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Sample records for mouse mus musculus

  1. Track recognition in 4 [mu]s by a systolic trigger processor using a parallel Hough transform

    SciTech Connect (OSTI)

    Klefenz, F.; Noffz, K.H.; Conen, W.; Zoz, R.; Kugel, A. . Lehrstuhl fuer Informatik V); Maenner, R. . Lehrstuhl fuer Informatik V Univ. Heidelberg . Interdisziplinaeres Zentrum fuer Wissenschaftliches Rechnen)

    1993-08-01

    A parallel Hough transform processor has been developed that identifies circular particle tracks in a 2D projection of the OPAL jet chamber. The high-speed requirements imposed by the 8 bunch crossing mode of LEP could be fulfilled by computing the starting angle and the radius of curvature for each well defined track in less than 4 [mu]s. The system consists of a Hough transform processor that determines well defined tracks, and a Euler processor that counts their number by applying the Euler relation to the thresholded result of the Hough transform. A prototype of a systolic processor has been built that handles one sector of the jet chamber. It consists of 35 [times] 32 processing elements that were loaded into 21 programmable gate arrays (XILINX). This processor runs at a clock rate of 40 MHz. It has been tested offline with about 1,000 original OPAL events. No deviations from the off-line simulation have been found. A trigger efficiency of 93% has been obtained. The prototype together with the associated drift time measurement unit has been installed at the OPAL detector at LEP and 100k events have been sampled to evaluate the system under detector conditions.

  2. Comparative mapping in the beige-satin region of mouse chromosome 13

    SciTech Connect (OSTI)

    Perou, C.M.; Pryor, R.; Kaplan, J.

    1997-01-15

    The proximal end of mouse chromosome (Chr) 13 contains regions conserved on human chromosomes 1q42-q44, 6p23-p21, and 7p22-p13. This region also contains mutations that may be models for human disease, including beige (human Chediak-Higashi syndrome). An interspecific backcross of SB/Le and Mus spretus mice was used to generate a molecular genetic linkage map of mouse chromosome 13 with an emphasis on the proximal region including beige (bg) and satin (sa). This map provides the gene order of the two phenotypic markers bg and sa relative to restriction fragment length polymorphisms and simple sequence length polymorphisms in 131 backcross animals. In parallel, we have created a physical map of the region using Nidogen (Nid) as a molecular starting point for cloning a YAC contig that was used to identify the beige gene. The physical map provides the fine-structure order of genes and anonymous DNA fragments that was not resolved by the genetic linkage mapping. The results show that the bg region of mouse Chr 13 is highly conserved on human Chr 1q42-q44 and provide a starting point for a complete functional analysis of the entire bg-sa interval. 37 refs., 4 figs., 1 tab.

  3. Mark Musculus named SAE fellow

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    ... in-cylinder processes of low-temperature combustion as well as new laser... for the heavy-duty, optical diesel engine project in the Combustion Research ...

  4. The mouse genome informatics and the mouse genome database

    SciTech Connect (OSTI)

    Maltais, L.J.; Blackburn, R.E.; Bradt, D.W.

    1994-09-01

    The Mouse Genome Database (MGD) is a centralized, comprehensive database of the mouse genome that includes genetic mapping data, comparative mapping data, gene descriptions, mutant phenotype descriptions, strains and allelic polymorphism data, inbred strain characteristics, physical mapping data, and molecular probes and clones data. Data in MGD are obtained from the published literature and by electronic transfer from laboratories working on large backcross panels of mice. MGD provides tools that enable the user to search the database, retrieve data, generate reports, analyze data, annotate records, and build genetic maps. The Encyclopedia of the Mouse Genome provides a graphic user interface to mouse genome data. It consists of software tools including: LinkMap, a graphic display of genetic linkage maps with the ability to magnify regions of high locus density: CytoMap, a graphic display of cytogenetic maps showing banded chromosomes with cytogenetic locations of genes and chromosomal aberrations; CATS, a catalog searching tool for text retrieval of mouse locus descriptions. These software tools provide access to the following data sets: Chromosome Committee Reports, MIT Genome Center data, GBASE reports, Mouse Locus Catalog (MLC), and Mouse Cytogenetic Mapping Data. The MGD is available to the scientific community through the World Wide Web (WWW) and Gopher. In addition GBASE can be accessed via the Internet.

  5. RAPID/Best Practices/Memorandums of Understanding (MOUs) | Open...

    Open Energy Info (EERE)

    Practices(Redirected from RAPIDBest PracticesMemorandums of Understanding (MOUs) for Interstate Transmission Projects)...

  6. Computational, Integrative, and Comparative Methods for the Elucidation of Genetic Coexpression Networks

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Baldwin, Nicole E.; Chesler, Elissa J.; Kirov, Stefan; Langston, Michael A.; Snoddy, Jay R.; Williams, Robert W.; Zhang, Bing

    2005-01-01

    Gene expression microarray data can be used for the assembly of genetic coexpression network graphs. Using mRNA samples obtained from recombinant inbred Mus musculus strains, it is possible to integrate allelic variation with molecular and higher-order phenotypes. The depth of quantitative genetic analysis of microarray data can be vastly enhanced utilizing this mouse resource in combination with powerful computational algorithms, platforms, and data repositories. The resulting network graphs transect many levels of biological scale. This approach is illustrated with the extraction of cliques of putatively co-regulated genes and their annotation using gene ontology analysis and cis -regulatory element discovery.more » The causal basis for co-regulation is detected through the use of quantitative trait locus mapping.« less

  7. Laboratory Memoranda of Understanding (MOUs) with Foreign Partners

    Broader source: Directives, Delegations, and Requirements [Office of Management (MA)]

    2012-05-14

    This memorandum establishes policy and procedures for any Memoranda of Understanding (MOUs) between DOE National Laboratories and any foreign entity, whether governmental or private.

  8. The Mouse House: a brief history of the ORNL mouse-genetics program, 1947-2009

    SciTech Connect (OSTI)

    Russell, Liane B

    2013-01-01

    The large mouse genetics program at the Oak Ridge National Lab is often re-membered chiefly for the germ-cell mutation-rate data it generated and their uses in estimating the risk of heritable radiation damage. In fact, it soon became a multi-faceted research effort that, over a period of almost 60 years, generated a wealth of information in the areas of mammalian mutagenesis, basic genetics (later enriched by molecular techniques), cytogenetics, reproductive biology, biochemistry of germ cells, and teratology. Research in the area of germ-cell mutagenesis explored the important physical and biological factors that affect the frequency and nature of induced mutations and made several unexpected discoveries, such as the major importance of the perigametic interval (the zygote stage) for the origin of spontaneous mutations and for the sensitivity to induced genetic change. Of practical value was the discovery that ethylnitrosourea was a supermutagen for point mutations, making high-efficiency mutagenesis in the mouse feasible worldwide. Teratogenesis findings resulted in recommendations still generally accepted in radiological practice. Studies supporting the mutagenesis research added whole bodies of information about mammalian germ-cell development and about molecular targets in germ cells. The early decision to not merely count but propagate genetic variants of all sorts made possible further discoveries, such as the Y-Chromosome s importance in mammalian sex determination and the identification of rare X-autosome translocations, which, in turn, led to the formulation of the single-active-X hypothesis and provided tools for studies of functional mosaicism for autosomal genes, male sterility, and chromosome-pairing mechanism. Extensive genetic and then molecular analyses of large numbers of induced specific-locus mutants resulted in fine-structure physical and correlated functional mapping of significant portions of the mouse genome and constituted a valuable source of mouse models for human genetic disorders.

  9. Insights from Human/Mouse genome comparisons

    SciTech Connect (OSTI)

    Pennacchio, Len A.

    2003-03-30

    Large-scale public genomic sequencing efforts have provided a wealth of vertebrate sequence data poised to provide insights into mammalian biology. These include deep genomic sequence coverage of human, mouse, rat, zebrafish, and two pufferfish (Fugu rubripes and Tetraodon nigroviridis) (Aparicio et al. 2002; Lander et al. 2001; Venter et al. 2001; Waterston et al. 2002). In addition, a high-priority has been placed on determining the genomic sequence of chimpanzee, dog, cow, frog, and chicken (Boguski 2002). While only recently available, whole genome sequence data have provided the unique opportunity to globally compare complete genome contents. Furthermore, the shared evolutionary ancestry of vertebrate species has allowed the development of comparative genomic approaches to identify ancient conserved sequences with functionality. Accordingly, this review focuses on the initial comparison of available mammalian genomes and describes various insights derived from such analysis.

  10. Transgenic Mouse Model of Chronic Beryllium Disease

    SciTech Connect (OSTI)

    Gordon, Terry

    2009-05-26

    Animal models provide powerful tools for dissecting dose-response relationships and pathogenic mechanisms and for testing new treatment paradigms. Mechanistic research on beryllium exposure-disease relationships is severely limited by a general inability to develop a sufficient chronic beryllium disease animal model. Discovery of the Human Leukocyte Antigen (HLA) - DPB1Glu69 genetic susceptibility component of chronic beryllium disease permitted the addition of this human beryllium antigen presentation molecule to an animal genome which may permit development of a better animal model for chronic beryllium disease. Using FVB/N inbred mice, Drs. Rubin and Zhu, successfully produced three strains of HLA-DPB1 Glu 69 transgenic mice. Each mouse strain contains a haplotype of the HLA-DPB1 Glu 69 gene that confers a different magnitude of odds ratio (OR) of risk for chronic beryllium disease: HLA-DPB1*0401 (OR = 0.2), HLA-DPB1*0201 (OR = 15), HLA-DPB1*1701 (OR = 240). In addition, Drs. Rubin and Zhu developed transgenic mice with the human CD4 gene to permit better transmission of signals between T cells and antigen presenting cells. This project has maintained the colonies of these transgenic mice and tested the functionality of the human transgenes.

  11. Control Board Digital Interface Input Devices Touchscreen, Trackpad, or Mouse?

    SciTech Connect (OSTI)

    Thomas A. Ulrich; Ronald L. Boring; Roger Lew

    2015-08-01

    The authors collaborated with a power utility to evaluate input devices for use in the human system interface (HSI) for a new digital Turbine Control System (TCS) at a nuclear power plant (NPP) undergoing a TCS upgrade. A standalone dynamic software simulation of the new digital TCS and a mobile kiosk were developed to conduct an input device study to evaluate operator preference and input device effectiveness. The TCS software presented the anticipated HSI for the TCS and mimicked (i.e., simulated) the turbine systems responses to operator commands. Twenty-four licensed operators from the two nuclear power units participated in the study. Three input devices were tested: a trackpad, mouse, and touchscreen. The subjective feedback from the survey indicates the operators preferred the touchscreen interface. The operators subjectively rated the touchscreen as the fastest and most comfortable input device given the range of tasks they performed during the study, but also noted a lack of accuracy for selecting small targets. The empirical data suggest the mouse input device provides the most consistent performance for screen navigation and manipulating on screen controls. The trackpad input device was both empirically and subjectively found to be the least effective and least desired input device.

  12. Janus Experiments: Data from Mouse Irradiation Experiments 1972 - 1989

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    The Janus Experiments, carried out at Argonne National Laboratory from 1972 to 1989 and supported by grants from the US Department of Energy, investigated the effects of neutron and gamma radiation on mouse tissues primarily from B6CF1 mice. 49,000 mice were irradiated: Death records were recorded for 42,000 mice; gross pathologies were recorded for 39,000 mice; and paraffin embedded tissues were preserved for most mice. Mouse record details type and source of radiation [gamma, neutrons]; dose and dose rate [including life span irradiation]; type and presence/absence of radioprotector treatment; tissue/animal morphology and pathology. Protracted low dose rate treatments, short term higher dose rate treatments, variable dose rates with a same total dose, etc. in some cases in conjunction with radioprotectors, were administered. Normal tissues, tumors, metastases were preserved. Standard tissues saved were : lung, liver, spleen, kidney, heart, any with gross lesions (including mammary glands, Harderian gland with eye, adrenal gland, gut, ovaries or testes, brain and pituitary, bone). Data are searchable and specimens can be obtained by request.

  13. The biology of novel animal genes: Mouse APEX gene knockout

    SciTech Connect (OSTI)

    MacInnes, M.; Altherr, M.R.; Ludwig, D.; Pedersen, R.; Mold, C.

    1997-07-01

    This is the final report of a one-year, Laboratory Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The controlled breeding of novel genes into mice, including the gene knockout (KO), or conversely by adding back transgenes provide powerful genetic technologies that together suffice to determine in large part the biological role(s) of novel genes. Inbred mouse remains the best understood and most useful mammalian experimental system available for tackling the biology of novel genes. The major mammalian apurinic/apyrimidinic (AP) endonuclease (APE), is involved in a key step in the repair of spontaneous and induced AP sites in DNA. Efficient repair of these lesions is imperative to prevent the stable incorporation of mutations into the cellular genome which may lead to cell death or transformation. Loss or modulation of base excison repair activity in vivo may elevate the spontaneous mutation rate in cells, and may lead to a substantial increase in the incidence of cancer. Despite extensive biochemical analysis, however, the significance of these individual APE functions in vivo has not been elucidated. Mouse embryonic stem (ES) cells heterozygous for a deletion mutation in APE have been generated and whole animals containing the APE mutation have been derived from these ES cells. Animals homozygous for the APE null mutation die early in gestation, underscoring the biological significance of this DNA repair gene.

  14. GATA-1 directly regulates Nanog in mouse embryonic stem cells

    SciTech Connect (OSTI)

    Li, Wen-Zhong; Ai, Zhi-Ying; Wang, Zhi-Wei; Chen, Lin-Lin; Guo, Ze-Kun; Zhang, Yong

    2015-09-25

    Nanog safeguards pluripotency in mouse embryonic stem cells (mESCs). Insight into the regulation of Nanog is important for a better understanding of the molecular mechanisms that control pluripotency of mESCs. In a silico analysis, we identify four GATA-1 putative binding sites in Nanog proximal promoter. The Nanog promoter activity can be significantly repressed by ectopic expression of GATA-1 evidenced by a promoter reporter assay. Mutation studies reveal that one of the four putative binding sites counts for GATA-1 repressing Nanog promoter activity. Direct binding of GATA-1 on Nanog proximal promoter is confirmed by electrophoretic mobility shift assay and chromatin immunoprecipitation. Our data provide new insights into the expanded regulatory circuitry that coordinates Nanog expression. - Highlights: • The Nanog proximal promoter conceives functional element for GATA-1. • GATA-1 occupies the Nanog proximal promoter in vitro and in vivo. • GATA-1 transcriptionally suppresses Nanog.

  15. A Systematic Analysis of a Deep Mouse Epididymal Sperm Proteome

    SciTech Connect (OSTI)

    Chauvin, Theodore; Xie, Fang; Liu, Tao; Nicora, Carrie D.; Yang, Feng; Camp, David G.; Smith, Richard D.; Roberts, Kenneth P.

    2012-12-21

    Spermatozoa are highly specialized cells that, when mature, are capable of navigating the female reproductive tract and fertilizing an oocyte. The sperm cell is thought to be largely quiescent in terms of transcriptional and translational activity. As a result, once it has left the male reproductive tract, the sperm cell is essentially operating with a static population of proteins. It is therefore theoretically possible to understand the protein networks contained in a sperm cell and to deduce its cellular function capabilities. To this end we have performed a proteomic analysis of mouse sperm isolated from the cauda epididymis and have confidently identified 2,850 proteins, which is the most comprehensive sperm proteome for any species reported to date. These proteins comprise many complete cellular pathways, including those for energy production via glycolysis, ?-oxidation and oxidative phosphorylation, protein folding and transport, and cell signaling systems. This proteome should prove a useful tool for assembly and testing of protein networks important for sperm function.

  16. The Crystal Structure of Mouse Exo70 Reveals Unique Features of the

    Office of Scientific and Technical Information (OSTI)

    Mammalian Exocyst (Journal Article) | SciTech Connect SciTech Connect Search Results Journal Article: The Crystal Structure of Mouse Exo70 Reveals Unique Features of the Mammalian Exocyst Citation Details In-Document Search Title: The Crystal Structure of Mouse Exo70 Reveals Unique Features of the Mammalian Exocyst Authors: Moore, Brian A. ; Robinson, Howard H. ; Xu, Zhaohui [1] ; Michigan-Med) [2] + Show Author Affiliations BNL ( Publication Date: 2015-08-24 OSTI Identifier: 1186909

  17. An Anisotropic Fluid-Solid Model of the Mouse Heart (Conference) | SciTech

    Office of Scientific and Technical Information (OSTI)

    Connect Conference: An Anisotropic Fluid-Solid Model of the Mouse Heart Citation Details In-Document Search Title: An Anisotropic Fluid-Solid Model of the Mouse Heart A critical challenge in biomechanical simulations is the spatial discretization of complex fluid-solid geometries created from imaging. This is especially important when dealing with Lagrangian interfaces, as there must be at a minimum both geometric and topological compatibility between fluid and solid phases, with exact

  18. A developmentally plastic adult mouse kidney cell line spontaneously generates multiple adult kidney structures

    SciTech Connect (OSTI)

    Webb, Carol F.; Ratliff, Michelle L.; Powell, Rebecca; Wirsig-Wiechmann, Celeste R.; Lakiza, Olga; Obara, Tomoko

    2015-08-07

    Despite exciting new possibilities for regenerative therapy posed by the ability to induce pluripotent stem cells, recapitulation of three-dimensional kidneys for repair or replacement has not been possible. ARID3a-deficient mouse tissues generated multipotent, developmentally plastic cells. Therefore, we assessed the adult mouse ARID3a−/− kidney cell line, KKPS5, which expresses renal progenitor surface markers as an alternative cell source for modeling kidney development. Remarkably, these cells spontaneously developed into multicellular nephron-like structures in vitro, and engrafted into immunocompromised medaka mesonephros, where they formed mouse nephron structures. These data implicate KKPS5 cells as a new model system for studying kidney development. - Highlights: • An ARID3a-deficient mouse kidney cell line expresses multiple progenitor markers. • This cell line spontaneously forms multiple nephron-like structures in vitro. • This cell line formed mouse kidney structures in immunocompromised medaka fish kidneys. • Our data identify a novel model system for studying kidney development.

  19. Interpretation of the Moessbauer Spectra of the Magnetic Nanoparticles in Mouse Spleen

    SciTech Connect (OSTI)

    Chuev, Mikhail A.; Cherepanov, Valery M.; Polikarpov, Mikhail A.; Panchenko, Vladislav Y.; Deyev, Sergey M.; Mischenko, Iliya N.; Nikitin, Maxim P.

    2010-12-02

    We have developed a stochastic model for description of relaxation effects in the system of homogeneously magnetized single-domain particles and applied the model to the analysis of Moessbauer spectra of magnetic nanoparticles (Chemicell ARA) and mouse spleen after i.v. injection into animals. We estimate that the fraction of exogenous iron in nanoparticles in the mouse spleen 3 months after injection was 0.27{+-}0.03. The spectra of the residual nanoparticles in the spleen had almost the same isomer shift but smaller mean hyperfine magnetic field values indicating decrease in the magnetic anisotropy energy (size) of the particles compared to the initial ones in the course of biodegradation. Concentration of ferritin-like iron was about three-fold higher than that in the spleen of untreated animals showing ferritin-like forms in the mouse spleen.

  20. Characterization of the Mouse Brain Proteome Using Global Proteomic Analysis Complemented with Cysteinyl-Peptide Enrichment

    SciTech Connect (OSTI)

    Wang, Haixing H.; Qian, Weijun; Chin, Mark H.; Petyuk, Vladislav A.; Barry, Richard C.; Liu, Tao; Gritsenko, Marina A.; Mottaz, Heather M.; Moore, Ronald J.; Camp, David G.; Khan, Arshad H.; Smith, Desmond; Smith, Richard D.

    2006-02-01

    Given the growing interest in applying genomic and proteomic approaches for studying the mammalian brain using mouse models, we hereby present for the first time a comprehensive characterization of the mouse brain proteome. Preparation of the whole brain sample incorporated a highly efficient cysteinyl-peptide enrichment (CPE) technique to complement a global enzymatic digestion method. Both the global and the cysteinyl-enriched peptide samples were analyzed by SCX fractionation coupled with reversed phase LC-MS/MS analysis. A total of 48,328 different peptides were confidently identified (>98% confidence level), covering 7792 non-redundant proteins (~34% of the predicted mouse proteome). 1564 and 1859 proteins were identified exclusively from the cysteinyl-peptide and the global peptide samples, respectively, corresponding to 25% and 31% improvements in proteome coverage compared to analysis of only the global peptide or cysteinyl-peptide samples. The identified proteins provide a broad representation of the mouse proteome with little bias evident due to protein pI, molecular weight, and/or cellular localization. Approximately 26% of the identified proteins with gene ontology (GO) annotations were membrane proteins, with 1447 proteins predicted to have transmembrane domains, and many of the membrane proteins were found to be involved in transport and cell signaling. The MS/MS spectrum count information for the identified proteins was used to provide a measure of relative protein abundances. The mouse brain peptide/protein database generated from this study represents the most comprehensive proteome coverage for the mammalian brain to date, and the basis for future quantitative brain proteomic studies using mouse models.

  1. Automated whole-genome multiple alignment of rat, mouse, and human

    SciTech Connect (OSTI)

    Brudno, Michael; Poliakov, Alexander; Salamov, Asaf; Cooper, Gregory M.; Sidow, Arend; Rubin, Edward M.; Solovyev, Victor; Batzoglou, Serafim; Dubchak, Inna

    2004-07-04

    We have built a whole genome multiple alignment of the three currently available mammalian genomes using a fully automated pipeline which combines the local/global approach of the Berkeley Genome Pipeline and the LAGAN program. The strategy is based on progressive alignment, and consists of two main steps: (1) alignment of the mouse and rat genomes; and (2) alignment of human to either the mouse-rat alignments from step 1, or the remaining unaligned mouse and rat sequences. The resulting alignments demonstrate high sensitivity, with 87% of all human gene-coding areas aligned in both mouse and rat. The specificity is also high: <7% of the rat contigs are aligned to multiple places in human and 97% of all alignments with human sequence > 100kb agree with a three-way synteny map built independently using predicted exons in the three genomes. At the nucleotide level <1% of the rat nucleotides are mapped to multiple places in the human sequence in the alignment; and 96.5% of human nucleotides within all alignments agree with the synteny map. The alignments are publicly available online, with visualization through the novel Multi-VISTA browser that we also present.

  2. Mapping of the NEP receptor tyrosine kinase gene to human chromosome 6p21.3 and mouse chromosome 17C

    SciTech Connect (OSTI)

    Edelhoff, S.; Disteche, C.M.; Sweetser, D.A.

    1995-01-01

    The mouse receptor tyrosine kinase (RTK) NEP, also called Ptk-3, is widely expressed, with high levels in proliferating neuroepithelia of mouse embryos. The recently described human discoidin domain receptor (DDR) has a predicted amino acid sequence 93% identical to that of murine NEP and may be its human homologue. We have mapped the gene encoding NEP in human and mouse by fluorescence in situ hybridization using a mouse cDNA probe. The NEP/Nep gene maps to human chromosome 6p21.3 and mouse chromosome 17C, respectively. This places the NEP/Nep gene at, or near, the major histocompatibility (MHC) locus-HLA in human and H2 in mouse, respectively. Based on its pattern of expression during development, NEP and Nep represent candidate genes for several MHC-linked developmental abnormalities in human and mouse. 19 refs., 1 fig.

  3. Human-mouse comparative genomics: successes and failures to reveal functional regions of the human genome

    SciTech Connect (OSTI)

    Pennacchio, Len A.; Baroukh, Nadine; Rubin, Edward M.

    2003-05-15

    Deciphering the genetic code embedded within the human genome remains a significant challenge despite the human genome consortium's recent success at defining its linear sequence (Lander et al. 2001; Venter et al. 2001). While useful strategies exist to identify a large percentage of protein encoding regions, efforts to accurately define functional sequences in the remaining {approx}97 percent of the genome lag. Our primary interest has been to utilize the evolutionary relationship and the universal nature of genomic sequence information in vertebrates to reveal functional elements in the human genome. This has been achieved through the combined use of vertebrate comparative genomics to pinpoint highly conserved sequences as candidates for biological activity and transgenic mouse studies to address the functionality of defined human DNA fragments. Accordingly, we describe strategies and insights into functional sequences in the human genome through the use of comparative genomics coupled wit h functional studies in the mouse.

  4. Sulfur mustard induces an endoplasmic reticulum stress response in the mouse ear vesicant model

    SciTech Connect (OSTI)

    Chang, Yoke-Chen; Wang, James D.; Svoboda, Kathy K.; Casillas, Robert P.; Laskin, Jeffrey D.; Gordon, Marion K.; Gerecke, Donald R.

    2013-04-15

    The endoplasmic reticulum (ER) stress response is a cell survival pathway upregulated when cells are under severe stress. Severely damaged mouse ear skin exposed to the vesicant, sulfur mustard (bis-2-chloroethyl sulfide, SM), resulted in increased expression of ER chaperone proteins that accompany misfolded and incorrectly made proteins targeted for degradation. Time course studies with SM using the mouse ear vesicant model (MEVM) showed progressive histopathologic changes including edema, separation of the epidermis from the dermis, persistent inflammation, upregulation of laminin ?2 (one of the chains of laminin-332, a heterotrimeric skin glycoprotein required for wound repair), and delayed wound healing from 24 h to 168 h post exposure. This was associated with time related increased expression of the cell survival ER stress marker, GRP78/BiP, and the ER stress apoptosis marker, GADD153/CHOP, suggesting simultaneous activation of both cell survival and non-mitochondrial apoptosis pathways. Dual immunofluorescence labeling of a keratinocyte migration promoting protein, laminin ?2 and GRP78/BIP, showed colocalization of the two molecules 72 h post exposure indicating that the laminin ?2 was misfolded after SM exposure and trapped within the ER. Taken together, these data show that ER stress is induced in mouse skin within 24 h of vesicant exposure in a defensive response to promote cell survival; however, it appears that this response is rapidly overwhelmed by the apoptotic pathway as a consequence of severe SM-induced injury. - Highlights: ? We demonstrated ER stress response in the mouse ear vesicant model. ? We described the asymmetrical nature of wound repair in the MEVM. ? We identified the distribution of various ER stress markers in the MEVM.

  5. Thalidomide induced early gene expression perturbations indicative of human embryopathy in mouse embryonic stem cells

    SciTech Connect (OSTI)

    Gao, Xiugong Sprando, Robert L.; Yourick, Jeffrey J.

    2015-08-15

    Developmental toxicity testing has traditionally relied on animal models which are costly, time consuming, and require the sacrifice of large numbers of animals. In addition, there are significant disparities between human beings and animals in their responses to chemicals. Thalidomide is a species-specific developmental toxicant that causes severe limb malformations in humans but not in mice. Here, we used microarrays to study transcriptomic changes induced by thalidomide in an in vitro model based on differentiation of mouse embryonic stem cells (mESCs). C57BL/6 mESCs were allowed to differentiate spontaneously and RNA was collected at 24, 48, and 72 h after exposure to 0.25 mM thalidomide. Global gene expression analysis using microarrays revealed hundreds of differentially expressed genes upon thalidomide exposure that were enriched in gene ontology (GO) terms and canonical pathways associated with embryonic development and differentiation. In addition, many genes were found to be involved in small GTPases-mediated signal transduction, heart development, and inflammatory responses, which coincide with clinical evidences and may represent critical embryotoxicities of thalidomide. These results demonstrate that transcriptomics in combination with mouse embryonic stem cell differentiation is a promising alternative model for developmental toxicity assessment. - Highlights: • Studied genomic changes in mouse embryonic stem cells upon thalidomide exposure • Identified gene expression changes that may represent thalidomide embryotoxicity • The toxicogenomic changes coincide well with known thalidomide clinical outcomes. • The mouse embryonic stem cell model is suitable for developmental toxicity testing. • The model has the potential for high-throughput screening of a multitude of compounds.

  6. Radiation-Induced Alterations in Mouse Brain Development Characterized by Magnetic Resonance Imaging

    SciTech Connect (OSTI)

    Gazdzinski, Lisa M.; Cormier, Kyle; Lu, Fred G.; Lerch, Jason P.; Department of Medical Biophysics, University of Toronto, Toronto ; Wong, C. Shun; Department of Medical Biophysics, University of Toronto, Toronto; Department of Radiation Oncology, University of Toronto, Toronto ; Nieman, Brian J.

    2012-12-01

    Purpose: The purpose of this study was to identify regions of altered development in the mouse brain after cranial irradiation using longitudinal magnetic resonance imaging (MRI). Methods and Materials: Female C57Bl/6 mice received a whole-brain radiation dose of 7 Gy at an infant-equivalent age of 2.5 weeks. MRI was performed before irradiation and at 3 time points following irradiation. Deformation-based morphometry was used to quantify volume and growth rate changes following irradiation. Results: Widespread developmental deficits were observed in both white and gray matter regions following irradiation. Most of the affected brain regions suffered an initial volume deficit followed by growth at a normal rate, remaining smaller in irradiated brains compared with controls at all time points examined. The one exception was the olfactory bulb, which in addition to an early volume deficit, grew at a slower rate thereafter, resulting in a progressive volume deficit relative to controls. Immunohistochemical assessment revealed demyelination in white matter and loss of neural progenitor cells in the subgranular zone of the dentate gyrus and subventricular zone. Conclusions: MRI can detect regional differences in neuroanatomy and brain growth after whole-brain irradiation in the developing mouse. Developmental deficits in neuroanatomy persist, or even progress, and may serve as useful markers of late effects in mouse models. The high-throughput evaluation of brain development enabled by these methods may allow testing of strategies to mitigate late effects after pediatric cranial irradiation.

  7. A mouse model of mitochondrial complex III dysfunction induced by myxothiazol

    SciTech Connect (OSTI)

    Davoudi, Mina; Kallijrvi, Jukka; Marjavaara, Sanna; Kotarsky, Heike; Hansson, Eva; Leven, Per; Fellman, Vineta

    2014-04-18

    Highlights: Reversible chemical inhibition of complex III in wild type mouse. Myxothiazol causes decreased complex III activity in mouse liver. The model is useful for therapeutic trials to improve mitochondrial function. - Abstract: Myxothiazol is a respiratory chain complex III (CIII) inhibitor that binds to the ubiquinol oxidation site Qo of CIII. It blocks electron transfer from ubiquinol to cytochrome b and thus inhibits CIII activity. It has been utilized as a tool in studies of respiratory chain function in in vitro and cell culture models. We developed a mouse model of biochemically induced and reversible CIII inhibition using myxothiazol. We administered myxothiazol intraperitoneally at a dose of 0.56 mg/kg to C57Bl/J6 mice every 24 h and assessed CIII activity, histology, lipid content, supercomplex formation, and gene expression in the livers of the mice. A reversible CIII activity decrease to 50% of control value occurred at 2 h post-injection. At 74 h only minor histological changes in the liver were found, supercomplex formation was preserved and no significant changes in the expression of genes indicating hepatotoxicity or inflammation were found. Thus, myxothiazol-induced CIII inhibition can be induced in mice for four days in a row without overt hepatotoxicity or lethality. This model could be utilized in further studies of respiratory chain function and pharmacological approaches to mitochondrial hepatopathies.

  8. Cell-autonomous progeroid changes in conditional mouse models for repair endonuclease XPG deficiency

    SciTech Connect (OSTI)

    Barnhoorn, Sander [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Uittenboogaard, Lieneke M. [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Jaarsma, Dick [Erasmus Univ. Medical Center, Rotterdam (The Netherlands). Dept. of Neuroscience.; Vermeij, Wilbert P. [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Tresini, Maria [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Weymaere, Michael [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Menoni, Herv [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Brandt, Renata M. C. [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; de Waard, Monique C. [VU Univ. Medical Center, Amsterdam (The Netherlands). Dept. of Intensive Care.; Botter, Sander M. [Uniklinik Balgrist, Zurich (Switzerland); Sarker, Altaf H. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division.; Jaspers, Nicolaas G. J. [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics; van der Horst, Gijsbertus T. J. [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; Cooper, Priscilla K. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division.; Hoeijmakers, Jan H. J. [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics.; van der Pluijm, Ingrid [Erasmus University Medical Center, Rotterdam (The Netherlands). Dept. of Genetics and Dept. of Vascular Surgery.; Niedernhofer, Laura J. [The Scripps Research Institute, San Diego, CA (United States)

    2014-10-09

    As part of the Nucleotide Excision Repair (NER) process, the endonuclease XPG is involved in repair of helix-distorting DNA lesions, but the protein has also been implicated in several other DNA repair systems, complicating genotype-phenotype relationship in XPG patients. Defects in XPG can cause either the cancer-prone condition xeroderma pigmentosum (XP) alone, or XP combined with the severe neurodevelopmental disorder Cockayne Syndrome (CS), or the infantile lethal cerebro-oculo-facio-skeletal (COFS) syndrome, characterized by dramatic growth failure, progressive neurodevelopmental abnormalities and greatly reduced life expectancy. Here, we present a novel (conditional) Xpg-/- mouse model whichin a C57BL6/FVB F1 hybrid genetic backgrounddisplays many progeroid features, including cessation of growth, loss of subcutaneous fat, kyphosis, osteoporosis, retinal photoreceptor loss, liver aging, extensive neurodegeneration, and a short lifespan of 45 months. We show that deletion of XPG specifically in the liver reproduces the progeroid features in the liver, yet abolishes the effect on growth or lifespan. In addition, specific XPG deletion in neurons and glia of the forebrain creates a progressive neurodegenerative phenotype that shows many characteristics of human XPG deficiency. Our findings therefore exclude that both the liver as well as the neurological phenotype are a secondary consequence of derailment in other cell types, organs or tissues (e.g. vascular abnormalities) and support a cell-autonomous origin caused by the DNA repair defect itself. In addition they allow the dissection of the complex aging process in tissue- and cell-type-specific components. Moreover, our data highlight the critical importance of genetic background in mouse aging studies, establish the Xpg-/- mouse as a valid model for the severe form of human XPG patients and segmental accelerated aging, and strengthen the link between DNA damage and aging.

  9. Characterization of a panel of somatic cell hybrids for regional mapping of the mouse X chromosome

    SciTech Connect (OSTI)

    Avner, P.; Arnaud, D.; Amar, L.; Cambrou, J.; Winking, H.; Russell, L.B.

    1987-08-01

    A panel of five hybrid cell lines containing mouse X chromosomes with various deletions has been obtained by fusing splenocytes from male mice carrying one of a series of reciprocal X-autosome translocations with the azaguanine-resistant Chinese hamster cell line CH3g. These hybrids have been extensively characterized by using the allozymes hypoxanthine/guanine phosphoribosyltransferase (encoded by the Hprt locus) and ..cap alpha..-galactosidase (Ags) and a series of 11 X-chromosome-specific DNA probes whose localization had been previously established by linkage studies. Such studies have established the genetic breakpoints of the T(X;12)13R1 and T(X;2)14R1 X-autosome translocations on the X chromosome and provided additional information as to the X-chromosome genetic breakpoints of the T(X;16)16H, T(X;4)7R1, and T(X;7)6R1 translocations. The data establish clearly that both the T(X;7)5RI and T(X;12)13R1 X-chromosome breakpoints are proximal to Hprt, the breakpoint of the former being more centromeric, lying as it does in the 9-centimorgan interval between the ornithine transcarbamoylase (Otc) and DXPas7 (M2C) loci. These five hybrid cell lines provide, with the previously characterized EBS4 hybrid cell line, a nested series of seven mapping intervals distributed along the length of the mouse X chromosome. Their characterization not only allows further correlation of the genetic and cytological X-chromosome maps but also should permit the rapid identification of DNA probes specific for particular regions of the mouse X chromosome.

  10. DNA repair decline during mouse spermiogenesis results in the accumulation of heritable DNA damage

    SciTech Connect (OSTI)

    Marchetti, Francesco; Marchetti, Francesco; Wryobek, Andrew J

    2008-02-21

    The post-meiotic phase of mouse spermatogenesis (spermiogenesis) is very sensitive to the genomic effects of environmental mutagens because as male germ cells form mature sperm they progressively lose the ability to repair DNA damage. We hypothesized that repeated exposures to mutagens during this repair-deficient phase result in the accumulation of heritable genomic damage in mouse sperm that leads to chromosomal aberrations in zygotes after fertilization. We used a combination of single or fractionated exposures to diepoxybutane (DEB), a component of tobacco smoke, to investigate how differential DNA repair efficiencies during the three weeks of spermiogenesis affected the accumulation of DEB-induced heritable damage in early spermatids (21-15 days before fertilization, dbf), late spermatids (14-8 dbf) and sperm (7- 1 dbf). Analysis of chromosomalaberrations in zygotic metaphases using PAINT/DAPI showed that late spermatids and sperm are unable to repair DEB-induced DNA damage as demonstrated by significant increases (P<0.001) in the frequencies of zygotes with chromosomal aberrations. Comparisons between single and fractionated exposures suggested that the DNA repair-deficient window during late spermiogenesis may be less than two weeks in the mouse and that during this repair-deficient window there is accumulation of DNA damage in sperm. Finally, the dose-response study in sperm indicated a linear response for both single and repeated exposures. These findings show that the differential DNA repair capacity of post-meioitic male germ cells has a major impact on the risk of paternally transmitted heritable damage and suggest that chronic exposures that may occur in the weeks prior to fertilization because of occupational or lifestyle factors (i.e, smoking) can lead to an accumulation of genetic damage in sperm and result in heritable chromosomal aberrations of paternal origin.

  11. DNA Repair Decline During Mouse Spermiogenesis Results in the Accumulation of Heritable DNA Damage

    SciTech Connect (OSTI)

    Marchetti, Francesco; Marchetti, Francesco; Wyrobek, Andrew J.

    2007-12-01

    The post-meiotic phase of mouse spermatogenesis (spermiogenesis) is very sensitive to the genomic effects of environmental mutagens because as male germ cells form mature sperm they progressively lose the ability to repair DNA damage. We hypothesized that repeated exposures to mutagens during this repair-deficient phase result in the accumulation of heritable genomic damage in mouse sperm that leads to chromosomal aberrations in zygotes after fertilization. We used a combination of single or fractionated exposures to diepoxybutane (DEB), a component of tobacco smoke, to investigate how differential DNA repair efficiencies during the three weeks of spermiogenesis affected the accumulation of DEB-induced heritable damage in early spermatids (21-15 days before fertilization, dbf), late spermatids (14-8 dbf) and sperm (7-1 dbf). Analysis of chromosomal aberrations in zygotic metaphases using PAINT/DAPI showed that late spermatids and sperm are unable to repair DEB-induced DNA damage as demonstrated by significant increases (P<0.001) in the frequencies of zygotes with chromosomal aberrations. Comparisons between single and fractionated exposures suggested that the DNA repair-deficient window during late spermiogenesis may be less than two weeks in the mouse and that during this repair-deficient window there is accumulation of DNA damage in sperm. Finally, the dose-response study in sperm indicated a linear response for both single and repeated exposures. These findings show that the differential DNA repair capacity of post-meioitic male germ cells has a major impact on the risk of paternally transmitted heritable damage and suggest that chronic exposures that may occur in the weeks prior to fertilization because of occupational or lifestyle factors (i.e, smoking) can lead to an accumulation of genetic damage in sperm and result in heritable chromosomal aberrations of paternal origin.

  12. Cell-autonomous progeroid changes in conditional mouse models for repair endonuclease XPG deficiency

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Barnhoorn, Sander; Uittenboogaard, Lieneke M.; Jaarsma, Dick; Vermeij, Wilbert P.; Tresini, Maria; Weymaere, Michael; Menoni, Hervé; Brandt, Renata M. C.; de Waard, Monique C.; Botter, Sander M.; et al

    2014-10-09

    As part of the Nucleotide Excision Repair (NER) process, the endonuclease XPG is involved in repair of helix-distorting DNA lesions, but the protein has also been implicated in several other DNA repair systems, complicating genotype-phenotype relationship in XPG patients. Defects in XPG can cause either the cancer-prone condition xeroderma pigmentosum (XP) alone, or XP combined with the severe neurodevelopmental disorder Cockayne Syndrome (CS), or the infantile lethal cerebro-oculo-facio-skeletal (COFS) syndrome, characterized by dramatic growth failure, progressive neurodevelopmental abnormalities and greatly reduced life expectancy. Here, we present a novel (conditional) Xpg-/- mouse model which—in a C57BL6/FVB F1 hybrid genetic background—displays manymore » progeroid features, including cessation of growth, loss of subcutaneous fat, kyphosis, osteoporosis, retinal photoreceptor loss, liver aging, extensive neurodegeneration, and a short lifespan of 4–5 months. We show that deletion of XPG specifically in the liver reproduces the progeroid features in the liver, yet abolishes the effect on growth or lifespan. In addition, specific XPG deletion in neurons and glia of the forebrain creates a progressive neurodegenerative phenotype that shows many characteristics of human XPG deficiency. Our findings therefore exclude that both the liver as well as the neurological phenotype are a secondary consequence of derailment in other cell types, organs or tissues (e.g. vascular abnormalities) and support a cell-autonomous origin caused by the DNA repair defect itself. In addition they allow the dissection of the complex aging process in tissue- and cell-type-specific components. Moreover, our data highlight the critical importance of genetic background in mouse aging studies, establish the Xpg-/- mouse as a valid model for the severe form of human XPG patients and segmental accelerated aging, and strengthen the link between DNA damage and aging.« less

  13. Pointright: a system to redirect mouse and keyboard control among multiple machines

    DOE Patents [OSTI]

    Johanson, Bradley E.; Winograd, Terry A.; Hutchins, Gregory M.

    2008-09-30

    The present invention provides a software system, PointRight, that allows for smooth and effortless control of pointing and input devices among multiple displays. With PointRight, a single free-floating mouse and keyboard can be used to control multiple screens. When the cursor reaches the edge of a screen it seamlessly moves to the adjacent screen and keyboard control is simultaneously redirected to the appropriate machine. Laptops may also redirect their keyboard and pointing device, and multiple pointers are supported simultaneously. The system automatically reconfigures itself as displays go on, go off, or change the machine they display.

  14. Arsenic- and cadmium-induced toxicogenomic response in mouse embryos undergoing neurulation

    SciTech Connect (OSTI)

    Robinson, Joshua F.; Yu, Xiaozhong; Moreira, Estefania G.; Hong, Sungwoo; Faustman, Elaine M.

    2011-01-15

    Arsenic (As) and cadmium (Cd) are well-characterized teratogens in animal models inducing embryotoxicity and neural tube defects (NTDs) when exposed during neurulation. Toxicological research is needed to resolve the specific biological processes and associated molecular pathways underlying metal-induced toxicity during this timeframe in gestational development. In this study, we investigated the dose-dependent effects of As and Cd on gene expression in C57BL/6J mouse embryos exposed in utero during neurulation (GD8) to identify significantly altered genes and corresponding biological processes associated with embryotoxicity. We quantitatively examined the toxicogenomic dose-response relationship at the gene level. Our results suggest that As and Cd induce dose-dependent gene expression alterations representing shared (cell cycle, response to UV, glutathione metabolism, RNA processing) and unique (alcohol/sugar metabolism) biological processes, which serve as robust indicators of metal-induced developmental toxicity and indicate underlying embryotoxic effects. Our observations also correlate well with previously identified impacts of As and Cd on specific genes associated with metal-induced toxicity (Cdkn1a, Mt1). In summary, we have identified in a quantitative manner As and Cd induced dose-dependent effects on gene expression in mouse embryos during a peak window of sensitivity to embryotoxicity and NTDs in the sensitive C57BL/6J strain.

  15. Sulforaphane induces phase II detoxication enzymes in mouse skin and prevents mutagenesis induced by a mustard gas analog

    SciTech Connect (OSTI)

    Abel, E.L.; Boulware, S.; Fields, T.; McIvor, E.; Powell, K.L.; DiGiovanni, J.; Vasquez, K.M.; MacLeod, M.C.

    2013-02-01

    Mustard gas, used in chemical warfare since 1917, is a mutagenic and carcinogenic agent that produces severe dermal lesions for which there are no effective therapeutics; it is currently seen as a potential terrorist threat to civilian populations. Sulforaphane, found in cruciferous vegetables, is known to induce enzymes that detoxify compounds such as the sulfur mustards that react through electrophilic intermediates. Here, we observe that a single topical treatment with sulforaphane induces mouse epidermal levels of the regulatory subunit of glutamate-cysteine ligase, the rate-limiting enzyme in glutathione biosynthesis, and also increases epidermal levels of reduced glutathione. Furthermore, a glutathione S-transferase, GSTA4, is also induced in mouse skin by sulforaphane. In an in vivo model in which mice are given a single mutagenic application of the sulfur mustard analog 2-(chloroethyl) ethyl sulfide (CEES), we now show that therapeutic treatment with sulforaphane abolishes the CEES-induced increase in mutation frequency in the skin, measured four days after exposure. Sulforaphane, a natural product currently in clinical trials, shows promise as an effective therapeutic against mustard gas. -- Highlights: ► Sulforaphane induces increased levels of glutathione in mouse skin. ► Sulforaphane induces increased levels of GSTA4 in mouse skin. ► Sulforaphane, applied after CEES-treatment, completely abolishes CEES-mutagenesis. ► The therapeutic effect may suggest a long biological half-life for CEES in vivo.

  16. Metastable primordial germ cell-like state induced from mouse embryonic stem cells by Akt activation

    SciTech Connect (OSTI)

    Yamano, Noriko; Kimura, Tohru; Watanabe-Kushima, Shoko; Shinohara, Takashi; Nakano, Toru; Department of Pathology, Medical School, Osaka University, 2-2 Yamada-oka, Suita, Osaka 565-0871

    2010-02-12

    Specification to primordial germ cells (PGCs) is mediated by mesoderm-induction signals during gastrulation. We found that Akt activation during in vitro mesodermal differentiation of embryonic stem cells (ESCs) generated self-renewing spheres with differentiation states between those of ESCs and PGCs. Essential regulators for PGC specification and their downstream germ cell-specific genes were expressed in the spheres, indicating that the sphere cells had commenced differentiation to the germ lineage. However, the spheres did not proceed to spermatogenesis after transplantation into testes. Sphere cell transfer to the original feeder-free ESC cultures resulted in chaotic differentiation. In contrast, when the spheres were cultured on mouse embryonic fibroblasts or in the presence of ERK-cascade and GSK3 inhibitors, reversion to the ESC-like state was observed. These results indicate that Akt signaling promotes a novel metastable and pluripotent state that is intermediate to those of ESCs and PGCs.

  17. A simple, low-cost, data logging pendulum built from a computer mouse

    SciTech Connect (OSTI)

    Gintautas, Vadas; Hubler, Alfred

    2009-01-01

    Lessons and homework problems involving a pendulum are often a big part of introductory physics classes and laboratory courses from high school to undergraduate levels. Although laboratory equipment for pendulum experiments is commercially available, it is often expensive and may not be affordable for teachers on fixed budgets, particularly in developing countries. We present a low-cost, easy-to-build rotary sensor pendulum using the existing hardware in a ball-type computer mouse. We demonstrate how this apparatus may be used to measure both the frequency and coefficient of damping of a simple physical pendulum. This easily constructed laboratory equipment makes it possible for all students to have hands-on experience with one of the most important simple physical systems.

  18. Automatic analysis of flow cytometric DNA histograms from irradiated mouse male germ cells

    SciTech Connect (OSTI)

    Lampariello, F.; Mauro, F.; Uccelli, R.; Spano, M.

    1989-01-01

    An automatic procedure for recovering the DNA content distribution of mouse irradiated testis cells from flow cytometric histograms is presented. First, a suitable mathematical model is developed, to represent the pattern of DNA content and fluorescence distribution in the sample. Then a parameter estimation procedure, based on the maximum likelihood approach, is constructed by means of an optimization technique. This procedure has been applied to a set of DNA histograms relative to different doses of 0.4-MeV neutrons and to different time intervals after irradiation. In each case, a good agreement between the measured histograms and the corresponding fits has been obtained. The results indicate that the proposed method for the quantitative analysis of germ cell DNA histograms can be usefully applied to the study of the cytotoxic and mutagenic action of agents of toxicological interest such as ionizing radiations.18 references.

  19. MONICA: a compact, portable dual gamma camera system for mouse whole-body imaging

    SciTech Connect (OSTI)

    Choyke, Peter L.; Xia, Wenze; Seidel, Jurgen; Kakareka, John W.; Pohida, Thomas J.; Milenic, Diane E.; Proffitt, James; Majewski, Stan; Weisenberger, Andrew G.; Green, Michael V.

    2010-04-01

    Introduction We describe a compact, portable dual-gamma camera system (named "MONICA" for MObile Nuclear Imaging CAmeras) for visualizing and analyzing the whole-body biodistribution of putative diagnostic and therapeutic single photon emitting radiotracers in animals the size of mice. Methods Two identical, miniature pixelated NaI(Tl) gamma cameras were fabricated and installed ?looking up? through the tabletop of a compact portable cart. Mice are placed directly on the tabletop for imaging. Camera imaging performance was evaluated with phantoms and field performance was evaluated in a weeklong In-111 imaging study performed in a mouse tumor xenograft model. Results Tc-99m performance measurements, using a photopeak energy window of 140 keV?10%, yielded the following results: spatial resolution (FWHM at 1 cm), 2.2 mm; sensitivity, 149 cps (counts per seconds)/MBq (5.5 cps/μCi); energy resolution (FWHM, full width at half maximum), 10.8%; count rate linearity (count rate vs. activity), r2=0.99 for 0?185 MBq (0?5 mCi) in the field of view (FOV); spatial uniformity, <3% count rate variation across the FOV. Tumor and whole-body distributions of the In-111 agent were well visualized in all animals in 5-min images acquired throughout the 168-h study period. Conclusion Performance measurements indicate that MONICA is well suited to whole-body single photon mouse imaging. The field study suggests that inter-device communications and user-oriented interfaces included in the MONICA design facilitate use of the system in practice. We believe that MONICA may be particularly useful early in the (cancer) drug development cycle where basic whole-body biodistribution data can direct future development of the agent under study and where logistical factors, e.g., limited imaging space, portability and, potentially, cost are important.

  20. Flavanone silibinin treatment attenuates nitrogen mustard-induced toxic effects in mouse skin

    SciTech Connect (OSTI)

    Jain, Anil K.; Tewari-Singh, Neera; Inturi, Swetha; Kumar, Dileep; Orlicky, David J.; Agarwal, Chapla; White, Carl W.; Agarwal, Rajesh

    2015-05-15

    Currently, there is no effective antidote to prevent skin injuries by sulfur mustard (SM) and nitrogen mustard (NM), which are vesicating agents with potential relevance to chemical warfare, terrorist attacks, or industrial/laboratory accidents. Our earlier report has demonstrated the therapeutic efficacy of silibinin, a natural flavanone, in reversing monofunctional alkylating SM analog 2-chloroethyl ethyl sulfide-induced toxic effects in mouse skin. To translate this effect to a bifunctional alkylating vesicant, herein, efficacy studies were carried out with NM. Topical application of silibinin (1 or 2 mg) 30 min after NM exposure on the dorsal skin of male SKH-1 hairless mice significantly decreased NM-induced toxic lesions at 24, 72 or 120 h post-exposure. Specifically, silibinin treatment resulted in dose-dependent reduction of NM-induced increase in epidermal thickness, dead and denuded epidermis, parakeratosis and microvesication. Higher silibinin dose also caused a 79% and 51%reversal in NM-induced increases in myeloperoxidase activity and COX-2 levels, respectively. Furthermore, silibinin completely prevented NM-induced H2A.X phosphorylation, indicating reversal of DNA damage which could be an oxidative DNA damage as evidenced by high levels of 8-oxodG in NM-exposed mouse skin that was significantly reversed by silibinin. Together, these findings suggest that attenuation of NM-induced skin injury by silibinin is due to its effects on the pathways associated with DNA damage, inflammation, vesication and oxidative stress. In conclusion, results presented here support the optimization of silibinin as an effective treatment of skin injury by vesicants. - Highlights: • Silibinin treatment attenuated nitrogen mustard (NM)-induced skin injury. • Silibinin affects pathways associated with DNA damage, inflammation and vesication. • The efficacy of silibinin could also be associated with oxidative stress. • These results support testing and optimization of silibinin against SM-induced skin injury.

  1. Strain-dependent Damage in Mouse Lung After Carbon Ion Irradiation

    SciTech Connect (OSTI)

    Moritake, Takashi; Proton Medical Research Center, University of Tsukuba, Tsukuba ; Fujita, Hidetoshi; Yanagisawa, Mitsuru; Nakawatari, Miyako; Imadome, Kaori; Nakamura, Etsuko; Iwakawa, Mayumi; Imai, Takashi

    2012-09-01

    Purpose: To examine whether inherent factors produce differences in lung morbidity in response to carbon ion (C-ion) irradiation, and to identify the molecules that have a key role in strain-dependent adverse effects in the lung. Methods and Materials: Three strains of female mice (C3H/He Slc, C57BL/6J Jms Slc, and A/J Jms Slc) were locally irradiated in the thorax with either C-ion beams (290 MeV/n, in 6 cm spread-out Bragg peak) or with {sup 137}Cs {gamma}-rays as a reference beam. We performed survival assays and histologic examination of the lung with hematoxylin-eosin and Masson's trichrome staining. In addition, we performed immunohistochemical staining for hyaluronic acid (HA), CD44, and Mac3 and assayed for gene expression. Results: The survival data in mice showed a between-strain variance after C-ion irradiation with 10 Gy. The median survival time of C3H/He was significantly shortened after C-ion irradiation at the higher dose of 12.5 Gy. Histologic examination revealed early-phase hemorrhagic pneumonitis in C3H/He and late-phase focal fibrotic lesions in C57BL/6J after C-ion irradiation with 10 Gy. Pleural effusion was apparent in C57BL/6J and A/J mice, 168 days after C-ion irradiation with 10 Gy. Microarray analysis of irradiated lung tissue in the three mouse strains identified differential expression changes in growth differentiation factor 15 (Gdf15), which regulates macrophage function, and hyaluronan synthase 1 (Has1), which plays a role in HA metabolism. Immunohistochemistry showed that the number of CD44-positive cells, a surrogate marker for HA accumulation, and Mac3-positive cells, a marker for macrophage infiltration in irradiated lung, varied significantly among the three mouse strains during the early phase. Conclusions: This study demonstrated a strain-dependent differential response in mice to C-ion thoracic irradiation. Our findings identified candidate molecules that could be implicated in the between-strain variance to early hemorrhagic pneumonitis after C-ion irradiation.

  2. Cell-specific oxidative stress and cytotoxicity after wildfire coarse particulate matter instillation into mouse lung

    SciTech Connect (OSTI)

    Williams, Keisha M.; Franzi, Lisa M.; Last, Jerold A.

    2013-01-01

    Our previous work has shown that coarse particulate matter (PM{sub 10-2.5}) from wildfire smoke is more toxic to lung macrophages on an equal dose (by mass) basis than coarse PM isolated from normal ambient air, as evidenced by decreased numbers of macrophages in lung lavage fluid 6 and 24 hours after PM instillation into mouse lungs in vivo and by cytotoxicity to a macrophage cell line observed directly in vitro. We hypothesized that pulmonary macrophages from mice instilled with wildfire coarse PM would undergo more cytotoxicity than macrophages from controls, and that there would be an increase in oxidative stress in their lungs. Cytotoxicity was quantified as decreased viable macrophages and increased percentages of dead macrophages in the bronchoalveolar lavage fluid (BALF) of mice instilled with wildfire coarse PM. At 1 hour after PM instillation, we observed both decreased numbers of viable macrophages and increased dead macrophage percentages as compared to controls. An increase in free isoprostanes, an indicator of oxidative stress, from control values of 28.1 3.2 pg/mL to 83.9 12.2 pg/mL was observed a half-hour after PM instillation. By 1 hour after PM instillation, isoprostane values had returned to 30.4 7.6 pg/mL, not significantly different from control concentrations. Lung sections from mice instilled with wildfire coarse PM showed rapid Clara cell responses, with decreased intracellular staining for the Clara cell secretory protein CCSP 1 hour after wildfire PM instillation. In conclusion, very rapid cytotoxicity occurs in pulmonary macrophages and oxidative stress responses are seen 0.51 hour after wildfire coarse PM instillation. These results define early cellular and biochemical events occurring in vivo and support the hypothesis that oxidative stress-mediated macrophage toxicity plays a key role in the initial response of the mouse lung to wildfire PM exposure. -- Highlights: ? We studied very early events (0.51 hour) after giving wildfire PM{sub 10-2.5} to mice. ? Wildfire PM{sub 10-2.5} rapidly kills lung macrophages in mice. ? Wildfire PM{sub 10-2.5} rapidly elicits oxidative stress in mice. ? Wildfire PM{sub 10-2.5} rapidly elicits Clara cell CCSP secretion in mice. ? Wildfire PM{sub 10-2.5} rapidly elicits TNF-? secretion into BALF in mice.

  3. PR-Set7 is degraded in a conditional Cul4A transgenic mouse model of lung cancer

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; Hsieh, David; Au, Alfred; Jablons, David M.; Li, Hui; You, Lian

    2015-06-01

    Background and objective. Maintenance of genomic integrity is essential to ensure normal organismal development and to prevent diseases such as cancer. PR-Set7 (also known as Set8) is a cell cycle regulated enzyme that catalyses monomethylation of histone 4 at Lys20 (H4K20me1) to promote chromosome condensation and prevent DNA damage. Recent studies show that CRL4CDT2-mediated ubiquitylation of PR-Set7 leads to its degradation during S phase and after DNA damage. This might occur to ensure appropriate changes in chromosome structure during the cell cycle or to preserve genome integrity after DNA damage. Methods. We developed a new model of lung tumor developmentmore » in mice harboring a conditionally expressed allele of Cul4A. We have therefore used a mouse model to demonstrate for the first time that Cul4A is oncogenic in vivo. With this model, staining of PR-Set7 in the preneoplastic and tumor lesions in AdenoCre-induced mouse lungs was performed. Meanwhile we identified higher protein level changes of γ-tubulin and pericentrin by IHC. Results. The level of PR-Set7 down-regulated in the preneoplastic and adenocarcinomous lesions following over-expression of Cul4A. We also identified higher levels of the proteins pericentrin and γ-tubulin in Cul4A mouse lungs induced by AdenoCre. Conclusion. PR-Set7 is a direct target of Cul4A for degradation and involved in the formation of lung tumors in the conditional Cul4A transgenic mouse model.« less

  4. 1H NMR Metabolomics Study of Metastatic Melanoma in C57BL/6J Mouse Spleen

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wang, Xuan; Hu, Mary Y.; Feng, Ju; Liu, Maili; Hu, Jian Z.

    2014-04-03

    Melanoma is a malignant tumor of melanocytes. Although extensive investigations have been done to study metabolic changes in primary melanoma in vivo and in vitro, little effort has been devoted to metabolic profiling of metastatic tumors in organs other than lymph nodes. In this work, NMR-based metabolomics combined with multivariate data analysis is used to study metastatic B16-F10 melanoma in C57BL/6J mouse spleen. Principal Component Analysis (PCA), an unsupervised multivariate data analysis method, is used to detect possible outliers, while Orthogonal Projection to Latent Structure (OPLS), a supervised multivariate data analysis method, is employed to find important metabolites responsible formore » discriminating the control and the melanoma groups. Two different strategies, i.e., spectral binning and spectral deconvolution, are used to reduce the original spectral data before statistical analysis. Spectral deconvolution is found to be superior for identifying a set of discriminatory metabolites between the control and the melanoma groups, especially when the sample size is small. OPLS results show that the melanoma group can be well separated from its control group. It is found that taurine, glutamate, aspartate, O-Phosphoethanolamine, niacinamide ,ATP, lipids and glycerol derivatives are decreased statistically and significantly while alanine, malate, xanthine, histamine, dCTP, GTP, thymidine, 2'-Deoxyguanosine are statistically and significantly elevated. These significantly changed metabolites are associated with multiple biological pathways and may be potential biomarkers for metastatic melanoma in spleen.« less

  5. X-ray scatter imaging of hepatocellular carcinoma in a mouse model using nanoparticle contrast agents

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Rand, Danielle; Derdak, Zoltan; Carlson, Rolf; Wands, Jack R.; Rose-Petruck, Christoph

    2015-10-29

    Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide and is almost uniformly fatal. Current methods of detection include ultrasound examination and imaging by CT scan or MRI; however, these techniques are problematic in terms of sensitivity and specificity, and the detection of early tumors (<1 cm diameter) has proven elusive. Better, more specific, and more sensitive detection methods are therefore urgently needed. Here we discuss the application of a newly developed x-ray imaging technique called Spatial Frequency Heterodyne Imaging (SFHI) for the early detection of HCC. SFHI uses x-rays scattered by an object to form anmore » image and is more sensitive than conventional absorption-based x-radiography. We show that tissues labeled in vivo with gold nanoparticle contrast agents can be detected using SFHI. We also demonstrate that directed targeting and SFHI of HCC tumors in a mouse model is possible through the use of HCC-specific antibodies. As a result, the enhanced sensitivity of SFHI relative to currently available techniques enables the x-ray imaging of tumors that are just a few millimeters in diameter and substantially reduces the amount of nanoparticle contrast agent required for intravenous injection relative to absorption-based x-ray imaging.« less

  6. X-ray scatter imaging of hepatocellular carcinoma in a mouse model using nanoparticle contrast agents

    SciTech Connect (OSTI)

    Rand, Danielle; Derdak, Zoltan; Carlson, Rolf; Wands, Jack R.; Rose-Petruck, Christoph

    2015-10-29

    Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide and is almost uniformly fatal. Current methods of detection include ultrasound examination and imaging by CT scan or MRI; however, these techniques are problematic in terms of sensitivity and specificity, and the detection of early tumors (<1 cm diameter) has proven elusive. Better, more specific, and more sensitive detection methods are therefore urgently needed. Here we discuss the application of a newly developed x-ray imaging technique called Spatial Frequency Heterodyne Imaging (SFHI) for the early detection of HCC. SFHI uses x-rays scattered by an object to form an image and is more sensitive than conventional absorption-based x-radiography. We show that tissues labeled in vivo with gold nanoparticle contrast agents can be detected using SFHI. We also demonstrate that directed targeting and SFHI of HCC tumors in a mouse model is possible through the use of HCC-specific antibodies. As a result, the enhanced sensitivity of SFHI relative to currently available techniques enables the x-ray imaging of tumors that are just a few millimeters in diameter and substantially reduces the amount of nanoparticle contrast agent required for intravenous injection relative to absorption-based x-ray imaging.

  7. Chromosomal mosaicism in mouse two-cell embryos after paternal exposure to acrylamide

    SciTech Connect (OSTI)

    Marchetti, Francesco; Bishop, Jack; Lowe, Xiu; Wyrobek, Andrew J

    2008-10-14

    Chromosomal mosaicism in human preimplantation embryos is a common cause ofspontaneous abortions, however, our knowledge of its etiology is limited. We used multicolor fluorescence in situ hybridization (FISH) painting to investigate whether paternally-transmitted chromosomal aberrations result in mosaicism in mouse 2-cell embryos. Paternal exposure to acrylamide, an important industrial chemical also found in tobacco smoke and generated during the cooking process of starchy foods, produced significant increases in chromosomally defective 2-cell embryos, however, the effects were transient primarily affecting the postmeiotic stages of spermatogenesis. Comparisons with our previous study of zygotes demonstrated similar frequencies of chromosomally abnormal zygotes and 2-cell embryos suggesting that there was no apparent selection against numerical or structural chromosomal aberrations. However, the majority of affected 2-cell embryos were mosaics showing different chromosomal abnormalities in the two blastomeric metaphases. Analyses of chromosomal aberrations in zygotes and 2-cell embryos showed a tendency for loss of acentric fragments during the first mitotic division ofembryogenesis, while both dicentrics and translocations apparently underwent propersegregation. These results suggest that embryonic development can proceed up to the end of the second cell cycle of development in the presence of abnormal paternal chromosomes and that even dicentrics can persist through cell division. The high incidence of chromosomally mosaic 2-cell embryos suggests that the first mitotic division of embryogenesis is prone to missegregation errors and that paternally-transmitted chromosomal abnromalities increase the risk of missegregation leading to embryonic mosaicism.

  8. DIFFERENTIAL SENSITIVITY OF MALE GERM CELLS TO MAINSTREAM AND SIDESTREAM TOBACCO SMOKE IN THE MOUSE

    SciTech Connect (OSTI)

    Polyzos, Aris; Schmid, Thomas Ernst; Pina-Guzman, Belem; Quintanilla-Vega, Betzabet; Marchetti, Francesco

    2009-03-13

    Cigarette smoking in men has been associated with increased chromosomal abnormalities in sperm and with increased risks for spontaneous abortions, birth defects and neonatal death. Little is known, however, about the reproductive consequences of paternal exposure to second-hand smoke. We used a mouse model to investigate the effects of paternal exposure to sidestream (SS) smoke, the main constituent of second-hand smoke, on the genetic integrity and function of sperm, and to determine whether male germ cells were equally sensitive to mainstream (MS) and SS smoke. A series of sperm DNA quality and reproductive endpoints were investigated after exposing male mice for two weeks to MS or SS smoke. Our results indicated that: (i) only SS smoke significantly affected sperm motility; (ii) only MS smoke induced DNA strand breaks in sperm; (iii) both MS and SS smoke increased sperm chromatin structure abnormalities; and (iv) MS smoke affected both fertilization and the rate of early embryonic development, while SS smoke affected fertilization only. These results show that MS and SS smoke have differential effects on the genetic integrity and function of sperm and provide further evidence that male exposure to second-hand smoke, as well as direct cigarette smoke, may diminish a couple's chance for a successful pregnancy and the birth of a healthy baby.

  9. Curine inhibits eosinophil activation and airway hyper-responsiveness in a mouse model of allergic asthma

    SciTech Connect (OSTI)

    Ribeiro-Filho, Jaime; Calheiros, Andrea Surrage; Vieira-de-Abreu, Adriana; Moraes de Carvalho, Katharinne Ingrid; Silva Mendes, Diego da; Melo, Christianne Bandeira; Martins, Marco Aurlio; Silva Dias, Celidarque da; Piuvezam, Mrcia Regina; and others

    2013-11-15

    Allergic asthma is a chronic inflammatory airway disease with increasing prevalence around the world. Current asthma therapy includes drugs that usually cause significant side effects, justifying the search for new anti-asthmatic drugs. Curine is a bisbenzylisoquinoline alkaloid that modulates calcium influx in many cell types; however, its anti-allergic and putative toxic effects remain to be elucidated. Our aim was to investigate the effects of curine on eosinophil activation and airway hyper-responsiveness (AHR) and to characterize its potential toxic effects. We used a mouse model of allergic asthma induced by sensitization and challenge with ovalbumin (OVA) to evaluate the anti-allergic effects of oral treatment with curine. The oral administration of curine significantly inhibited eosinophilic inflammation, eosinophil lipid body formation and AHR in animals challenged with OVA compared with animals in the untreated group. The curine treatment also reduced eotaxin and IL-13 production triggered by OVA. Verapamil, a calcium channel antagonist, had similar anti-allergic properties, and curine pre-treatment inhibited the calcium-induced tracheal contractile response ex-vivo, suggesting that the mechanism by which curine exerts its effects is through the inhibition of a calcium-dependent response. A toxicological evaluation showed that orally administered curine did not significantly alter the biochemical, hematological, behavioral and physical parameters measured in the experimental animals compared with saline-treated animals. In conclusion, curine showed anti-allergic activity through mechanisms that involve inhibition of IL-13 and eotaxin and of Ca{sup ++} influx, without inducing evident toxicity and as such, has the potential for the development of anti-asthmatic drugs. - Highlights: Curine is a bisbenzylisoquinoline alkaloid from Chondrodendron platyphyllum. Curine inhibits eosinophil influx and activation and airway hyper-responsiveness. Curine mechanisms involve inhibition of Ca{sup 2+} influx, and IL-13 and eotaxin secretion. No significant toxicity was observed in mice orally treated with curine for 7 days. Curine has the potential for the development of anti-asthmatic drugs.

  10. Methoxychlor reduces estradiol levels by altering steroidogenesis and metabolism in mouse antral follicles in vitro

    SciTech Connect (OSTI)

    Basavarajappa, Mallikarjuna S. Craig, Zelieann R. Hernandez-Ochoa, Isabel Paulose, Tessie Leslie, Traci C. Flaws, Jodi A.

    2011-06-15

    The organochlorine pesticide methoxychlor (MXC) is a known endocrine disruptor that affects adult rodent females by causing reduced fertility, persistent estrus, and ovarian atrophy. Since MXC is also known to target antral follicles, the major producer of sex steroids in the ovary, the present study was designed to test the hypothesis that MXC decreases estradiol (E{sub 2}) levels by altering steroidogenic and metabolic enzymes in the antral follicles. To test this hypothesis, antral follicles were isolated from CD-1 mouse ovaries and cultured with either dimethylsulfoxide (DMSO) or MXC. Follicle growth was measured every 24 h for 96 h. In addition, sex steroid hormone levels were measured using enzyme-linked immunosorbent assays (ELISA) and mRNA expression levels of steroidogenic enzymes as well as the E{sub 2} metabolic enzyme Cyp1b1 were measured using qPCR. The results indicate that MXC decreased E{sub 2}, testosterone, androstenedione, and progesterone (P{sub 4}) levels compared to DMSO. In addition, MXC decreased expression of aromatase (Cyp19a1), 17{beta}-hydroxysteroid dehydrogenase 1 (Hsd17b1), 17{alpha}-hydroxylase/17,20-lyase (Cyp17a1), 3{beta} hydroxysteroid dehydrogenase 1 (Hsd3b1), cholesterol side-chain cleavage (Cyp11a1), steroid acute regulatory protein (Star), and increased expression of Cyp1b1 enzyme levels. Thus, these data suggest that MXC decreases steroidogenic enzyme levels, increases metabolic enzyme expression and this in turn leads to decreased sex steroid hormone levels. - Highlights: > MXC inhibits steroidogenesis > MXC inhibits steroidogenic enzymes > MXC induces metabolic enzymes

  11. Isoniazid suppresses antioxidant response element activities and impairs adipogenesis in mouse and human preadipocytes

    SciTech Connect (OSTI)

    Chen, Yanyan; Xue, Peng; Hou, Yongyong; Zhang, Hao; Zheng, Hongzhi; Zhou, Tong; Qu, Weidong; Teng, Weiping; Zhang, Qiang; Andersen, Melvin E.; Pi, Jingbo

    2013-12-15

    Transcriptional signaling through the antioxidant response element (ARE), orchestrated by the Nuclear factor E2-related factor 2 (Nrf2), is a major cellular defense mechanism against oxidative or electrophilic stress. Here, we reported that isoniazid (INH), a widely used antitubercular drug, displays a substantial inhibitory property against ARE activities in diverse mouse and human cells. In 3T3-L1 preadipocytes, INH concentration-dependently suppressed the ARE-luciferase reporter activity and mRNA expression of various ARE-dependent antioxidant genes under basal and oxidative stressed conditions. In keeping with our previous findings that Nrf2-ARE plays a critical role in adipogenesis by regulating expression of CCAAT/enhancer-binding protein ? (C/EBP?) and peroxisome proliferator-activated receptor ? (PPAR?), suppression of ARE signaling by INH hampered adipogenic differentiation of 3T3-L1 cells and human adipose-derived stem cells (ADSCs). Following adipogenesis induced by hormonal cocktails, INH-treated 3T3-L1 cells and ADSCs displayed significantly reduced levels of lipid accumulation and attenuated expression of C/EBP? and PPAR?. Time-course studies in 3T3-L1 cells revealed that inhibition of adipogenesis by INH occurred in the early stage of terminal adipogenic differentiation, where reduced expression of C/EBP? and C/EBP? was observed. To our knowledge, the present study is the first to demonstrate that INH suppresses ARE signaling and interrupts with the transcriptional network of adipogenesis, leading to impaired adipogenic differentiation. The inhibition of ARE signaling may be a potential underlying mechanism by which INH attenuates cellular antioxidant response contributing to various complications. - Highlights: Isoniazid suppresses ARE-mediated transcriptional activity. Isoniazid inhibits adipogenesis in preadipocytes. Isoniazid suppresses adipogenic gene expression during adipogenesis.

  12. Comparative mapping of DNA markers from the familial Alzheimer disease and Down syndrome regions of human chromosome 21 to mouse chromosomes 16 and 17

    SciTech Connect (OSTI)

    Cheng, S.V.; Nadeau, J.H.; Tanzi, R.E.; Watkins, P.C.; Jagadesh, J.; Taylor, B.A.; Haines, J.L.; Sacchi, N.; Gusella, J.F. )

    1988-08-01

    Mouse trisomy 16 has been proposed as an animal model of Down syndrome (DS), since this chromosome contains homologues of several loci from the q22 band of human chromosome 21. The recent mapping of the defect causing familial Alzheimer disease (FAD) and the locus encoding the Alzheimer amyloid {beta} precursor protein (APP) to human chromosome 21 has prompted a more detailed examination of the extent of conservation of this linkage group between the two species. Using anonymous DNA probes and cloned genes from human chromosome 21 in a combination of recombinant inbred and interspecific mouse backcross analyses, the authors have established that the linkage group shared by mouse chromosome 16 includes not only the critical DS region of human chromosome 21 but also the APP gene and FAD-linked markers. Extending from the anonymous DNA locus D21S52 to ETS2, the linkage map of six loci spans 39% recombination in man but only 6.4% recombination in the mouse. A break in synteny occurs distal to ETS2, with the homologue of the human marker D21S56 mapping to mouse chromosome 17. Conservation of the linkage relationships of markers in the FAD region suggests that the murine homologue of the FAD locus probably maps to chromosome 16 and that detailed comparison of the corresponding region in both species could facilitate identification of the primary defect in this disorder. The break in synteny between the terminal portion of human chromosome 21 and mouse chromosome 16 indicates, however, that mouse trisomy 16 may not represent a complete model of DS.

  13. Inducible bilirubin oxidase: A novel function for the mouse cytochrome P450 2A5

    SciTech Connect (OSTI)

    Abu-Bakar, A'edah; Arthur, Dionne Maioha; Cooperative Research Centre for Contamination Assessment and Remediation of the Environment, Adelaide ; Aganovic, Simona; Ng, Jack C.; Cooperative Research Centre for Contamination Assessment and Remediation of the Environment, Adelaide ; Lang, Matti A.; Department of Pharmaceutical Biosciences, Uppsala University, Biomedical Centre, Box 578, S-751 23 Uppsala

    2011-11-15

    We have previously shown that bilirubin (BR), a breakdown product of haem, is a strong inhibitor and a high affinity substrate of the mouse cytochrome P450 2A5 (CYP2A5). The antioxidant BR, which is cytotoxic at high concentrations, is potentially useful in cellular protection against oxygen radicals if its intracellular levels can be strictly controlled. The mechanisms that regulate cellular BR levels are still obscure. In this paper we provide preliminary evidence for a novel function of CYP2A5 as hepatic 'BR oxidase'. A high-performance liquid chromatography/electrospray ionisation mass spectrometry screening showed that recombinant yeast microsomes expressing the CYP2A5 oxidise BR to biliverdin, as the main metabolite, and to three other smaller products with m/z values of 301, 315 and 333. The metabolic profile is significantly different from that of chemical oxidation of BR. In chemical oxidation the smaller products were the main metabolites. This suggests that the enzymatic reaction is selective, towards biliverdin production. Bilirubin treatment of primary hepatocytes increased the CYP2A5 protein and activity levels with no effect on the corresponding mRNA. Co-treatment with cycloheximide (CHX), a protein synthesis inhibitor, resulted in increased half-life of the CYP2A5 compared to cells treated only with CHX. Collectively, the observations suggest that the CYP2A5 is potentially an inducible 'BR oxidase' where BR may accelerate its own metabolism through stabilization of the CYP2A5 protein. It is possible that this metabolic pathway is potentially part of the machinery controlling intracellular BR levels in transient oxidative stress situations, in which high amounts of BR are produced. -- Highlights: Black-Right-Pointing-Pointer CYP2A5 metabolizes bilirubin to biliverdin and dipyrroles. Black-Right-Pointing-Pointer Bilirubin increased the hepatic CYP2A5 protein and activity levels. Black-Right-Pointing-Pointer Bilirubin does not change the hepatic CYP2A5 mRNA levels. Black-Right-Pointing-Pointer Co-treatment with a protein synthesis inhibitor prolongs CYP2A5 half-life. Black-Right-Pointing-Pointer CYP2A5 is potentially an inducible bilirubin oxidase.

  14. Transient inhibition of cell proliferation does not compromise self-renewal of mouse embryonic stem cells

    SciTech Connect (OSTI)

    Wang, Ruoxing; Guo, Yan-Lin

    2012-10-01

    Embryonic stem cells (ESCs) have unlimited capacity for self-renewal and can differentiate into various cell types when induced. They also have an unusual cell cycle control mechanism driven by constitutively active cyclin dependent kinases (Cdks). In mouse ESCs (mESCs). It is proposed that the rapid cell proliferation could be a necessary part of mechanisms that maintain mESC self-renewal and pluripotency, but this hypothesis is not in line with the finding in human ESCs (hESCs) that the length of the cell cycle is similar to differentiated cells. Therefore, whether rapid cell proliferation is essential for the maintenance of mESC state remains unclear. We provide insight into this uncertainty through chemical intervention of mESC cell cycle. We report here that inhibition of Cdks with olomoucine II can dramatically slow down cell proliferation of mESCs with concurrent down-regulation of cyclin A, B and E, and the activation of the Rb pathway. However, mESCs display can recover upon the removal of olomoucine II and are able to resume normal cell proliferation without losing self-renewal and pluripotency, as demonstrated by the expression of ESC markers, colony formation, embryoid body formation, and induced differentiation. We provide a mechanistic explanation for these observations by demonstrating that Oct4 and Nanog, two major transcription factors that play critical roles in the maintenance of ESC properties, are up-regulated via de novo protein synthesis when the cells are exposed to olomoucine II. Together, our data suggest that short-term inhibition of cell proliferation does not compromise the basic properties of mESCs. -- Highlights: Black-Right-Pointing-Pointer Inhibition of Cdks slows down mESCs proliferation. Black-Right-Pointing-Pointer mESCs display remarkable recovery capacity from short-term cell cycle interruption. Black-Right-Pointing-Pointer Short-term cell cycle interruption does not compromise mESC self-renewal. Black-Right-Pointing-Pointer Oct4 and Nanog are up-regulated via de novo synthesis by cell cycle interruption.

  15. UVB light upregulates prostaglandin synthases and prostaglandin receptors in mouse keratinocytes

    SciTech Connect (OSTI)

    Black, Adrienne T.; Gray, Joshua P.; Shakarjian, Michael P.; Mishin, Vladimir; Laskin, Debra L.; Heck, Diane E.; Laskin, Jeffrey D.

    2008-10-01

    Prostaglandins belong to a class of cyclic lipid-derived mediators synthesized from arachidonic acid via COX-1, COX-2 and various prostaglandin synthases. Members of this family include prostaglandins such as PGE{sub 2}, PGF{sub 2{alpha}}, PGD{sub 2} and PGI{sub 2} (prostacyclin) as well as thromboxane. In the present studies we analyzed the effects of UVB on prostaglandin production and prostaglandin synthase expression in primary cultures of undifferentiated and calcium-differentiated mouse keratinocytes. Both cell types were found to constitutively synthesize PGE{sub 2}, PGD{sub 2} and the PGD{sub 2} metabolite PGJ{sub 2}. Twenty-four hours after treatment with UVB (25 mJ/cm{sup 2}), production of PGE{sub 2} and PGJ{sub 2} increased, while PGD{sub 2} production decreased. This was associated with increased expression of COX-2 mRNA and protein. UVB (2.5-25 mJ/cm{sup 2}) also caused marked increases in mRNA expression for the prostanoid synthases PGDS, mPGES-1, mPGES-2, PGFS and PGIS, as well as expression of receptors for PGE{sub 2} (EP1 and EP2), PGD{sub 2} (DP and CRTH2) and prostacyclin (IP). UVB was more effective in inducing COX-2 and DP in differentiated cells and EP1 and IP in undifferentiated cells. UVB readily activated keratinocyte PI-3-kinase (PI3K)/Akt, JNK and p38 MAP signaling pathways which are known to regulate COX-2 expression. While inhibition of PI3K suppressed UVB-induced mPGES-1 and CRTH2 expression, JNK inhibition suppressed mPGES-1, PGIS, EP2 and CRTH2, and p38 kinase inhibition only suppressed EP1 and EP2. These data indicate that UVB modulates expression of prostaglandin synthases and receptors by distinct mechanisms. Moreover, both the capacity of keratinocytes to generate prostaglandins and their ability to respond to these lipid mediators are stimulated by exposure to UVB.

  16. Suppression of somatic expansion delays the onset of pathophysiology in a mouse model of Huntington’s Disease

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Budworth, Helen; Harris, Faye R.; Williams, Paul; Lee, Do Yup; Holt, Amy; Pahnke, Jens; Szczesny, Bartosz; Acevedo-Torres, Karina; Ayala-Peña, Sylvette; McMurray, Cynthia T.; et al

    2015-08-06

    Huntington’s Disease (HD) is caused by inheritance of a single disease-length allele harboring an expanded CAG repeat, which continues to expand in somatic tissues with age. The inherited disease allele expresses a toxic protein, and whether further somatic expansion adds to toxicity is unknown. We have created an HD mouse model that resolves the effects of the inherited and somatic expansions. We show here that suppressing somatic expansion substantially delays the onset of disease in littermates that inherit the same disease-length allele. Furthermore, a pharmacological inhibitor, XJB-5-131, inhibits the lengthening of the repeat tracks, and correlates with rescue of motormore » decline in these animals. The results provide evidence that pharmacological approaches to offset disease progression are possible.« less

  17. Suppression of somatic expansion delays the onset of pathophysiology in a mouse model of Huntington’s Disease

    SciTech Connect (OSTI)

    Budworth, Helen; Harris, Faye R.; Williams, Paul; Lee, Do Yup; Holt, Amy; Pahnke, Jens; Szczesny, Bartosz; Acevedo-Torres, Karina; Ayala-Peña, Sylvette; McMurray, Cynthia T.; McKinnon, Peter

    2015-08-06

    Huntington’s Disease (HD) is caused by inheritance of a single disease-length allele harboring an expanded CAG repeat, which continues to expand in somatic tissues with age. The inherited disease allele expresses a toxic protein, and whether further somatic expansion adds to toxicity is unknown. We have created an HD mouse model that resolves the effects of the inherited and somatic expansions. We show here that suppressing somatic expansion substantially delays the onset of disease in littermates that inherit the same disease-length allele. Furthermore, a pharmacological inhibitor, XJB-5-131, inhibits the lengthening of the repeat tracks, and correlates with rescue of motor decline in these animals. The results provide evidence that pharmacological approaches to offset disease progression are possible.

  18. The crystal structure of a partial mouse Notch-1 ankyrin domain: Repeats 4 through 7 preserve an ankyrin fold

    SciTech Connect (OSTI)

    Lubman, Olga Y.; Kopan, Raphael; Waksman, Gabriel; Korolev, Sergey (Birbeck); (St. Louis-MED); (WU-MED)

    2010-07-20

    Folding and stability of proteins containing ankyrin repeats (ARs) is of great interest because they mediate numerous protein-protein interactions involved in a wide range of regulatory cellular processes. Notch, an ankyrin domain containing protein, signals by converting a transcriptional repression complex into an activation complex. The Notch ANK domain is essential for Notch function and contains seven ARs. Here, we present the 2.2 {angstrom} crystal structure of ARs 4-7 from mouse Notch 1 (m1ANK). These C-terminal repeats were resistant to degradation during crystallization, and their secondary and tertiary structures are maintained in the absence of repeats 1-3. The crystallized fragment adopts a typical ankyrin fold including the poorly conserved seventh AR, as seen in the Drosophila Notch ANK domain (dANK). The structural preservation and stability of the C-terminal repeats shed a new light onto the mechanism of hetero-oligomeric assembly during Notch-mediated transcriptional activation.

  19. A Novel mouse model of enhanced proteostasis: Full-length human heat shock factor 1 transgenic mice

    SciTech Connect (OSTI)

    Pierce, Anson; Barshop Institute for Longevity and Aging Studies, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, 78229; The Department of Veteran's Affairs, South Texas Veterans Health Care System, San Antonio, Texas, 78284 ; Wei, Rochelle; Halade, Dipti; Yoo, Si-Eun; Barshop Institute for Longevity and Aging Studies, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, 78229 ; Ran, Qitao; Richardson, Arlan; Barshop Institute for Longevity and Aging Studies, The University of Texas Health Science Center at San Antonio, San Antonio, Texas, 78229; The Department of Veteran's Affairs, South Texas Veterans Health Care System, San Antonio, Texas, 78284

    2010-11-05

    Research highlights: {yields} Development of mouse overexpressing native human HSF1 in all tissues including CNS. {yields} HSF1 overexpression enhances heat shock response at whole-animal and cellular level. {yields} HSF1 overexpression protects from polyglutamine toxicity and favors aggresomes. {yields} HSF1 overexpression enhances proteostasis at the whole-animal and cellular level. -- Abstract: The heat shock response (HSR) is controlled by the master transcriptional regulator heat shock factor 1 (HSF1). HSF1 maintains proteostasis and resistance to stress through production of heat shock proteins (HSPs). No transgenic model exists that overexpresses HSF1 in tissues of the central nervous system (CNS). We generated a transgenic mouse overexpressing full-length non-mutant HSF1 and observed a 2-4-fold increase in HSF1 mRNA and protein expression in all tissues studied of HSF1 transgenic (HSF1{sup +/0}) mice compared to wild type (WT) littermates, including several regions of the CNS. Basal expression of HSP70 and 90 showed only mild tissue-specific changes; however, in response to forced exercise, the skeletal muscle HSR was more elevated in HSF1{sup +/0} mice compared to WT littermates and in fibroblasts following heat shock, as indicated by levels of inducible HSP70 mRNA and protein. HSF1{sup +/0} cells elicited a significantly more robust HSR in response to expression of the 82 repeat polyglutamine-YFP fusion construct (Q82YFP) and maintained proteasome-dependent processing of Q82YFP compared to WT fibroblasts. Overexpression of HSF1 was associated with fewer, but larger Q82YFP aggregates resembling aggresomes in HSF1{sup +/0} cells, and increased viability. Therefore, our data demonstrate that tissues and cells from mice overexpressing full-length non-mutant HSF1 exhibit enhanced proteostasis.

  20. Differential gene expression profiling of mouse skin after sulfur mustard exposure: Extended time response and inhibitor effect

    SciTech Connect (OSTI)

    Gerecke, Donald R. Chen Minjun; Isukapalli, Sastry S.; Gordon, Marion K.; Chang, Y.-C.; Tong Weida; Androulakis, Ioannis P.; Georgopoulos, Panos G.

    2009-01-15

    Sulfur mustard (HD, SM), is a chemical warfare agent that within hours causes extensive blistering at the dermal-epidermal junction of skin. To better understand the progression of SM-induced blistering, gene expression profiling for mouse skin was performed after a single high dose of SM exposure. Punch biopsies of mouse ears were collected at both early and late time periods following SM exposure (previous studies only considered early time periods). The biopsies were examined for pathological disturbances and the samples further assayed for gene expression profiling using the Affymetrix microarray analysis system. Principal component analysis and hierarchical cluster analysis of the differently expressed genes, performed with ArrayTrack showed clear separation of the various groups. Pathway analysis employing the KEGG library and Ingenuity Pathway Analysis (IPA) indicated that cytokine-cytokine receptor interaction, cell adhesion molecules (CAMs), and hematopoietic cell lineage are common pathways affected at different time points. Gene ontology analysis identified the most significantly altered biological processes as the immune response, inflammatory response, and chemotaxis; these findings are consistent with other reported results for shorter time periods. Selected genes were chosen for RT-PCR verification and showed correlations in the general trends for the microarrays. Interleukin 1 beta was checked for biological analysis to confirm the presence of protein correlated to the corresponding microarray data. The impact of a matrix metalloproteinase inhibitor, MMP-2/MMP-9 inhibitor I, against SM exposure was assessed. These results can help in understanding the molecular mechanism of SM-induced blistering, as well as to test the efficacy of different inhibitors.

  1. Dioxin exposure reduces the steroidogenic capacity of mouse antral follicles mainly at the level of HSD17B1 without altering atresia

    SciTech Connect (OSTI)

    Karman, Bethany N., E-mail: bklement@illinois.edu; Basavarajappa, Mallikarjuna S., E-mail: mbshivapur@gmail.com; Hannon, Patrick, E-mail: phannon2@illinois.edu; Flaws, Jodi A., E-mail: jflaws@illinois.edu

    2012-10-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent ovarian toxicant. Previously, we demonstrated that in vitro TCDD (1 nM) exposure decreases production/secretion of the sex steroid hormones progesterone (P4), androstenedione (A4), testosterone (T), and 17?-estradiol (E2) in mouse antral follicles. The purpose of this study was to determine the mechanism by which TCDD inhibits steroidogenesis. Specifically, we examined the effects of TCDD on the steroidogenic enzymes, atresia, and the aryl hydrocarbon receptor (AHR) protein. TCDD exposure for 48 h increased levels of A4, without changing HSD3B1 protein, HSD17B1 protein, estrone (E1), T or E2 levels. Further, TCDD did not alter atresia ratings compared to vehicle at 48 h. TCDD, however, did down regulate the AHR protein at 48 h. TCDD exposure for 96 h decreased transcript levels for Cyp11a1, Cyp17a1, Hsd17b1, and Cyp19a1, but increased Hsd3b1 transcript. TCDD exposure particularly lowered both Hsd17b1 transcript and HSD17B1 protein. However, TCDD exposure did not affect levels of E1 in the media nor atresia ratings at 96 h. TCDD, however, decreased levels of the proapoptotic factor Bax. Collectively, these data suggest that TCDD exposure causes a major block in the steroidogenic enzyme conversion of A4 to T and E1 to E2 and that it regulates apoptotic pathways, favoring survival over death in antral follicles. Finally, the down?regulation of the AHR protein in TCDD exposed follicles persisted at 96 h, indicating that the activation and proteasomal degradation of this receptor likely plays a central role in the impaired steroidogenic capacity and altered apoptotic pathway of exposed antral follicles. -- Highlights: ? TCDD disrupts steroidogenic enzymes in mouse antral follicles. ? TCDD particularly affects the HSD17B1 enzyme in mouse antral follicles. ? TCDD does not affect atresia ratings in mouse antral follicles. ? TCDD decreases levels of the proapoptitic factor Bax in mouse antral follicles. ? TCDD down regulates the AHR protein in mouse antral follicles.

  2. 2,6-Dithiopurine, a nucleophilic scavenger, protects against mutagenesis in mouse skin treated in vivo with 2-(chloroethyl) ethyl sulfide, a mustard gas analog

    SciTech Connect (OSTI)

    Boulware, Stephen; Fields, Tammy; McIvor, Elizabeth; Powell, K. Leslie; Abel, Erika L.; Vasquez, Karen M.; MacLeod, Michael C.

    2012-09-01

    Sulfur mustard [bis(2-chloroethyl)sulfide, SM] is a well-known DNA-damaging agent that has been used in chemical warfare since World War I, and is a weapon that could potentially be used in a terrorist attack on a civilian population. Dermal exposure to high concentrations of SM produces severe, long-lasting burns. Topical exposure to high concentrations of 2-(chloroethyl) ethyl sulfide (CEES), a monofunctional analog of SM, also produces severe skin lesions in mice. Utilizing a genetically engineered mouse strain, Big Blue, that allows measurement of mutation frequencies in mouse tissues, we now show that topical treatment with much lower concentrations of CEES induces significant dose- and time-dependent increases in mutation frequency in mouse skin; the mutagenic exposures produce minimal toxicity as determined by standard histopathology and immunohistochemical analysis for cytokeratin 6 and the DNA-damage induced phosphorylation of histone H2AX (γ-H2AX). We attempted to develop a therapeutic that would inhibit the CEES-induced increase in mutation frequency in the skin. We observe that multi-dose, topical treatment with 2,6-dithiopurine (DTP), a known chemical scavenger of CEES, beginning 1 h post-exposure to CEES, completely abolishes the CEES-induced increase in mutation frequency. These findings suggest the possibility that DTP, previously shown to be non-toxic in mice, may be useful as a therapeutic agent in accidental or malicious human exposures to SM. -- Highlights: ► 200 mM 2-(chloroethyl) ethyl sulfide (CEES) induces mutations in mouse skin. ► This dose of CEES is not overtly toxic, as assayed by histopathology. ► 2,6-Dithiopurine (DTP), applied after CEES-treatment, abolishes CEES-mutagenesis. ► This supports the idea that sulfur mustards exhibit long biological half-lives.

  3. Glomerular-specific imprinting of the mouse Gs{alpha} gene: How does this relate to hormone resistance in Albright hereditary osteodystrophy?

    SciTech Connect (OSTI)

    Williamson, C.M.; Dutton, E.R.; Seymour, A.

    1996-09-01

    The gene for alpha-stimulating guanine-nucleotide binding polypeptide, Gnas, has been considered as a candidate for the imprinting effects ascribed to distal mouse Chromosome (Chr) 2. Its human homologue (GNAS1) appears, from clinical and biochemical studies of patients with Albright hereditary ostodystrophy, to be paternally imprinted. GNAS1 maps to 20q13, a region that shows linkage conservation with distal mouse Chr 2. We have mapped Gnas within the imprinting region on distal Chr 2 by linkage analysis. To establish if Gnas is imprinted, we have looked for expression differences in tissues taken from mice carrying maternal duplication/paternal deficiency for distal Chr 2 (MatDp2) and its reciprocal (PatDp2). RNA in situ hybridization revealed high levels of Gnas mRNA in glomeruli of PatDp2 embryos at late gestation and lower levels in glomeruli of MatDp2 embryos. These results strongly suggest that Gnas is maternally imprinted and suggest that the mouse gene may be imprinted in a manner opposite the predicted in human. 42 refs., 4 figs.

  4. A novel rabbit anti-hepatocyte growth factor monoclonal neutralizing antibody inhibits tumor growth in prostate cancer cells and mouse xenografts

    SciTech Connect (OSTI)

    Yu, Yanlan; Chen, Yicheng; Ding, Guoqing; Wang, Mingchao; Wu, Haiyang; Xu, Liwei; Rui, Xuefang; Zhang, Zhigen

    2015-08-14

    The hepatocyte growth factor and its receptor c-Met are correlated with castration-resistance in prostate cancer. Although HGF has been considered as an attractive target for therapeutic antibodies, the lack of cross-reactivity of monoclonal antibodies with human/mouse HGFs is a major obstacle in preclinical developments. We generated a panel of anti-HGF RabMAbs either blocking HGF/c-Met interaction or inhibiting c-Met phosphorylation. We selected one RabMAb with mouse cross-reactivity and demonstrated that it blocked HGF-stimulated downstream activation in PC-3 and DU145 cells. Anti-HGF RabMAb inhibited not only the growth of PC-3 cells but also HGF-dependent proliferation in HUVECs. We further demonstrated the efficacy and potency of the anti-HGF RabMAb in tumor xenograft mice models. Through these in vitro and in vivo experiments, we explored a novel therapeutic antibody for advanced prostate cancer. - Highlights: • HGF is an attractive target for castration-refractory prostate cancer. • We generated and characterized a panel of anti-HGF rabbit monoclonal antibodies. • More than half of these anti-HGF RabMAbs was cross-reactive with mouse HGF. • Anti-HGF RabMAb blocks HGF-stimulated phosphorylation and cell growth in vitro. • Anti-HGF RabMAb inhibits tumor growth and angiogenesis in xenograft mice.

  5. Histone deacetylase inhibitor valproic acid promotes the induction of pluripotency in mouse fibroblasts by suppressing reprogramming-induced senescence stress

    SciTech Connect (OSTI)

    Zhai, Yingying; Chen, Xi; Yu, Dehai; Li, Tao; Cui, Jiuwei; Wang, Guanjun; Hu, Ji-Fan; Li, Wei

    2015-09-10

    Histone deacetylase inhibitor valproic acid (VPA) has been used to increase the reprogramming efficiency of induced pluripotent stem cell (iPSC) from somatic cells, yet the specific molecular mechanisms underlying this effect is unknown. Here, we demonstrate that reprogramming with lentiviruses carrying the iPSC-inducing factors (Oct4-Sox2-Klf4-cMyc, OSKM) caused senescence in mouse fibroblasts, establishing a stress barrier for cell reprogramming. Administration of VPA protected cells from reprogramming-induced senescent stress. Using an in vitro pre-mature senescence model, we found that VPA treatment increased cell proliferation and inhibited apoptosis through the suppression of the p16/p21 pathway. In addition, VPA also inhibited the G2/M phase blockage derived from the senescence stress. These findings highlight the role of VPA in breaking the cell senescence barrier required for the induction of pluripotency. - Highlights: • Histone deacetylase inhibitor valproic acid enhances iPSC induction. • Valproic acid suppresses reprogramming-induced senescence stress. • Valproic acid downregulates the p16/p21 pathway in reprogramming. • This study demonstrates a new mechanistic role of valproic acid in enhancing reprogramming.

  6. Metastatic Melanoma Induced Metabolic Changes in C57BL/6J Mouse Stomach Measured by 1H NMR Spectroscopy

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Hu, M; Wang, Xiliang

    2014-12-05

    Melanoma is a malignant tumor of melanocytes with high capability of invasion and rapid metastasis to other organs. Malignant melanoma is the most common metastatic malignancy found in gastrointestinal tract (GI). To the best of our knowledge, previous studies of melanoma in gastrointestinal tract are all clinical case reports. In this work, 1H NMR-based metabolomics approach is used to investigate the metabolite profiles differences of stomach tissue extracts of metastatic B16-F10 melanoma in C57BL/6J mouse and search for specific metabolite biomarker candidates. Principal Component Analysis (PCA), an unsupervised multivariate data analysis method, is used to detect possible outliers, while Orthogonalmore » Projection to Latent Structure (OPLS), a supervised multivariate data analysis method, is employed to evaluate important metabolites responsible for discriminating the control and the melanoma groups. Both PCA and OPLS results reveal that the melanoma group can be well separated from its control group. Among the 50 identified metabolites, it is found that the concentrations of 19 metabolites are statistically and significantly changed with the levels of O-phosphocholine and hypoxanthine down-regulated while the levels of isoleucine, leucine, valine, isobutyrate, threonine, cadaverine, alanine, glutamate, glutamine, methionine, citrate, asparagine, tryptophan, glycine, serine, uracil, and formate up-regulated in the melanoma group. These significantly changed metabolites are associated with multiple biological pathways and may be potential biomarkers for metastatic melanoma in stomach.« less

  7. Fourier Transform Infrared Imaging Showing Reduced Unsaturated Lipid Content in the Hippocampus of a mouse Model of Alzheimer's Disease

    SciTech Connect (OSTI)

    Leskovjan, A.C.; Kretlow, A.; Miller, L.M.

    2010-04-01

    Polyunsaturated fatty acids are essential to brain functions such as membrane fluidity, signal transduction, and cell survival. It is also thought that low levels of unsaturated lipid in the brain may contribute to Alzheimer's disease (AD) risk or severity. However, it is not known how accumulation of unsaturated lipids is affected in different regions of the hippocampus, which is a central target of AD plaque pathology, during aging. In this study, we used Fourier transform infrared imaging (FTIRI) to visualize the unsaturated lipid content in specific regions of the hippocampus in the PSAPP mouse model of AD as a function of plaque formation. Specifically, the unsaturated lipid content was imaged using the olefinic {double_bond}CH stretching mode at 3012 cm{sup -1}. The axonal, dendritic, and somatic layers of the hippocampus were examined in the mice at 13, 24, 40, and 56 weeks old. Results showed that lipid unsaturation in the axonal layer was significantly increased with normal aging in control (CNT) mice (p < 0.01) but remained low and relatively constant in PSAPP mice. Thus, these findings indicate that unsaturated lipid content is reduced in hippocampal white matter during amyloid pathogenesis and that maintaining unsaturated lipid content early in the disease may be critical in avoiding progression of the disease.

  8. Crystal Structure of Staphylococcal Enterotoxin G (SEG) in Complex with a Mouse T-cell Receptor Beta Chain

    SciTech Connect (OSTI)

    Fernandez, M.M.; Robinson, H.; Cho, S.; De Marzi, M. C.; Kerzic, M. C.; Mariuzza, R. A.; Malchiodi, E. L.

    2011-01-14

    Superantigens (SAgs) are bacterial or viral toxins that bind MHC class II (MHC-II) molecules and T-cell receptor (TCR) in a nonconventional manner, inducing T-cell activation that leads to inflammatory cytokine production, which may result in acute toxic shock. In addition, the emerging threat of purpura fulminans and community-associated meticillin-resistant Staphylococcus aureus emphasizes the importance of a better characterization of SAg binding to their natural ligands that may allow the development of reagents to neutralize their action. The three-dimensional structure of the complex between a mouse TCR {beta} chain (mV{beta}8.2) and staphylococcal enterotoxin G (SEG) at 2.0 {angstrom} resolution revealed a binding site that does not conserve the 'hot spots' present in mV{beta}8.2-SEC2, mV{beta}8.2-SEC3, mV{beta}8.2-SEB, and mV{beta}8.2-SPEA complexes. Analysis of the mV{beta}8.2-SEG interface allowed us to explain the higher affinity of this complex compared with the others, which may account for the early activation of T-cells bearing mV{beta}8.2 by SEG. This mode of interaction between SEG and mV{beta}8.2 could be an adaptive advantage to bestow on the pathogen a faster rate of colonization of the host.

  9. Metabolite signatures in hydrophilic extracts of mouse lungs exposed to cigarette smoke revealed by 1H NMR metabolomics investigation

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Hu, Jian Z.; Wang, Xuan; Feng, Ju; Webb-Robertson, Bobbie-Jo M.; Waters, Katrina M.; Tilton, Susan C.; Pounds, Joel G.; Corley, Richard A.; Liu, Maili; Hu, Mary Y.

    2015-05-12

    Herein, 1H-NMR metabolomics are carried out to evaluate the changes of metabolites in lungs of mice exposed to cigarette smoke. It is found that the concentrations of adenosine derivatives (i.e. ATP, ADP and AMP), inosine and uridine are significantly fluctuated in the lungs of mice exposed to cigarette smoke compared with those of controls regardless the mouse is obese or regular weight. The decreased ATP, ADP, AMP and elevated inosine predict that the deaminases in charge of adenosine derivatives to inosine derivatives conversion are altered in lungs of mice exposed to cigarette smoke. Transcriptional analysis reveals that the concentrations ofmore » adenosine monophosphate deaminase and adenosine deaminase are different in the lungs of mice exposed to cigarette smoke, confirming the prediction from metabolomics studies. We also found, for the first time, that the ratio of glycerophosphocholine (GPC) to phosphocholine (PC) is significantly increased in the lungs of obese mice compared with regular weight mice. The ratio of GPC/PC is further elevated in the lungs of obese group by cigarette smoke exposure. Since GPC/PC ratio is a known biomarker for cancer, these results may suggest that obese group is more susceptible to lung cancer when exposed to cigarette smoke.« less

  10. Metastatic Melanoma Induced Metabolic Changes in C57BL/6J Mouse Stomach Measured by 1H NMR Spectroscopy

    SciTech Connect (OSTI)

    Hu, M; Wang, Xiliang

    2014-12-05

    Melanoma is a malignant tumor of melanocytes with high capability of invasion and rapid metastasis to other organs. Malignant melanoma is the most common metastatic malignancy found in gastrointestinal tract (GI). To the best of our knowledge, previous studies of melanoma in gastrointestinal tract are all clinical case reports. In this work, 1H NMR-based metabolomics approach is used to investigate the metabolite profiles differences of stomach tissue extracts of metastatic B16-F10 melanoma in C57BL/6J mouse and search for specific metabolite biomarker candidates. Principal Component Analysis (PCA), an unsupervised multivariate data analysis method, is used to detect possible outliers, while Orthogonal Projection to Latent Structure (OPLS), a supervised multivariate data analysis method, is employed to evaluate important metabolites responsible for discriminating the control and the melanoma groups. Both PCA and OPLS results reveal that the melanoma group can be well separated from its control group. Among the 50 identified metabolites, it is found that the concentrations of 19 metabolites are statistically and significantly changed with the levels of O-phosphocholine and hypoxanthine down-regulated while the levels of isoleucine, leucine, valine, isobutyrate, threonine, cadaverine, alanine, glutamate, glutamine, methionine, citrate, asparagine, tryptophan, glycine, serine, uracil, and formate up-regulated in the melanoma group. These significantly changed metabolites are associated with multiple biological pathways and may be potential biomarkers for metastatic melanoma in stomach.

  11. Final Report of project entitled "A metabolomics and mouse models approach to study inflammatory and immune responses to radiation"

    SciTech Connect (OSTI)

    Fornace, Albert J.; Li, Henghong

    2013-12-02

    The three-year project entitled ?A Metabolomics and Mouse Models Approach to Study Inflammatory and Immune Responses to Radiation? was initiated in September 2009. The overall objectives of this project were to investigate the acute and persistent effects of low dose radiation on T cell lymphocyte function and physiology, as well the contributions of these cells to radiation-induced inflammatory responses. Inflammation after ionizing radiation (IR), even at low doses, may impact a variety of disease processes, including infectious disease, cardiovascular disease, cancer, and other potentially inflammatory disorders. There were three overall specific aims: 1. To investigate acute and persistent effects of low dose radiation on T cell subsets and function; 2. A genetic approach with mouse models to investigate p38 MAPK pathways that are involved in radiation-induced inflammatory signaling; 3. To investigate the effect of radiation quality on the inflammatory response. We have completed the work proposed in these aims. Below are our major accomplishments: ? Our data show that T cells from low dose irradiated animals have lower proliferation potency and cytokine production upon T cell receptor (TCR) stimulation. This effect was observed as early as 4 hours after radiation, and lasted up to two weeks. ? Using our ultraperformance liquid chromatography coupled with highly sensitive time-of-flight mass spectrometry (UPLC-QTOF) metabolomics method, we demonstrated the global changes of metabolites in T cells upon TCR stimulation in a time-dependent pattern. ? We found that the TCR activation induced metabolome changes are remarkably altered in a dose-dependent manner after radiation. At a dose of 0.5 Gy and above, IR mitigated TCR activation induced metabolome changes while at the dose of as low as 0.1Gy IR had a mild stimulatory effect on some of the metabolome changes. ? We revealed the mechanism for how radiation affects T cell activation by showing that the energy supply pathways in activated T cells are greatly compromised after radiation. ? We demonstrated that low dose ionizing radiation has a variety of effects on different T cell subsets, and p38 plays an important role in these effects. ? The study with low dose proton radiation shows similar effects on T cell proliferation upon TCR activation. Our dose rate study with proton radiation indicates that at low dose rates, proton exposure has less detrimental effects on T cell activation. ? We have one published paper and several manuscripts submitted or in preparation. ? We presented our findings at multiple DOE low dose program workshops, RRS annual meetings and other conferences. Our project is the first to apply a cutting-edge metabolomics approach to study the effects of radiation on immune cell function. Our findings demonstrate that metabolomics is a powerful method, which not only has higher sensitivity than the classical immune cell biology endpoints, but also helps to reveal the underlying mechanisms providing evidence that T cell activation is a metabolically dynamic process. Our T cell subset study sheds light on the effects of radiation on different T cell subsets and relevant signaling pathways mediating these effects. We have proved that our metabolomics platform and the T cell subset differentiation methods are useful and informative approaches for investigation and assessment of immune cell function after radiation. Our mechanistic findings on metabolic pathways may help to identify potential targets for intervention.

  12. Studies of Secondary Melanoma on C57BL/6J Mouse Liver Using 1H NMR Metabolomics

    SciTech Connect (OSTI)

    Feng, Ju; Isern, Nancy G.; Burton, Sarah D.; Hu, Jian Z.

    2013-10-31

    NMR metabolomics, consisting of solid state high resolution (hr) magic angle spinning (MAS) 1H NMR (1H hr-MAS), liquid state high resolution 1H-NMR, and principal components analysis (PCA) has been used to study secondary metastatic B16-F10 melanoma in C57BL/6J mouse liver . The melanoma group can be differentiated from its control group by PCA analysis of the absolute concentrations or by the absolute peak intensities of metabolites from either 1H hr-MAS NMR data on intact liver tissues or liquid state 1H-NMR spectra on liver tissue extracts. In particular, we found that the absolute concentrations of alanine, glutamate, creatine, creatinine, fumarate and cholesterol are elevated in the melanoma group as compared to controls, while the absolute concentrations of succinate, glycine, glucose, and the family of linear lipids including long chain fatty acids, total choline and acylglycerol are decreased. The ratio of glycerophosphocholine to phosphocholine is increased by about 1.5 fold in the melanoma group, while the absolute concentration of total choline is actually lower in melanoma mice. These results suggest the following picture in secondary melanoma metastasis: Linear lipid levels are decreased by beta oxidation in the melanoma group, which contributes to an increase in the synthesis of cholesterol, and also provides an energy source input for TCA cycle. These findings suggest a link between lipid oxidation, the TCA cycle and the hypoxia-inducible factors (HIF) signal pathway in tumor metastases. Thus this study indicates that the metabolic profile derived from NMR analysis can provide a valuable bio-signature of malignancy and cell hypoxia in metastatic melanoma.

  13. BENZO[a]PYRENE DIOL EPOXIDE PERTURBATION OF CELL CYCLE KINETICS OF SYNCHRONIZED MOUSE LIVER EPITHELIAL CELLS

    SciTech Connect (OSTI)

    Pearlman, A.L.; Navsky, B.N.; Bartholomew, J.C

    1980-07-01

    A cell cycle synchronization system is described for the analysis of the perturbation of cell cycle kinetics and the cycle-phase specificity of chemicals and other agents. We used the system to study the effects of ({+-})r-7, t-8-dihydroxy-t-9, 10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BaP diol epoxide) upon the cell cycle of mouse liver epithelial cells(NMuLi). BaP diol epoxide(0.6 uM) was added to replated cultures of NMuLi cells that had been synchronized in various stages of the cell cycle by centrifugal elutriation. DNA histograms were obtained by flow cytometry as a function of time after replating. The data were analyzed by a computer modeling routine and reduced to a few graphs illustrating the 'net effects' of the BaP diol epoxide relative to controls. BaP diol epoxide slowed S-phase traversal in all samples relative to their respective control. Traversal through G{sub 2}M was also slowed by at least 50%. BaP diol epoxide had no apparent effect upon G{sub 1} traversal by cycling cells, but delayed the recruitment of quiescent G{sub 0} cells by about 2 hrs. The methods described constitute a powerful new approach for probing the cell cycle effects of a wide variety of agents. The present system appears to be extremely sensitive and capable of characterizing the action of agents on each phase of the cell cycle. The methods are automatable and would allow for the assay and possible differential characterization of mutagens and carcinogens.

  14. Flavin-containing monooxygenase-3: Induction by 3-methylcholanthrene and complex regulation by xenobiotic chemicals in hepatoma cells and mouse liver

    SciTech Connect (OSTI)

    Celius, Trine; Pansoy, Andrea; Matthews, Jason; Okey, Allan B.; Henderson, Marilyn C.; Krueger, Sharon K.; Williams, David E.

    2010-08-15

    Flavin-containing monooxygenases often are thought not to be inducible but we recently demonstrated aryl hydrocarbon receptor (AHR)-dependent induction of FMO mRNAs in mouse liver by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) (Celius et al., Drug Metab Dispos 36:2499, 2008). We now evaluated FMO induction by other AHR ligands and xenobiotic chemicals in vivo and in mouse Hepa1c1c7 hepatoma cells (Hepa-1). In mouse liver, 3-methylcholanthrene (3MC) induced FMO3 mRNA 8-fold. In Hepa-1 cells, 3MC and benzo[a]pyrene (BaP) induced FMO3 mRNA > 30-fold. Induction by 3MC and BaP was AHR dependent but, surprisingly, the potent AHR agonist, TCDD, did not induce FMO3 mRNA in Hepa-1 cells nor did chromatin immunoprecipitation assays detect recruitment of AHR or ARNT to Fmo3 regulatory elements after exposure to 3MC in liver or in Hepa-1 cells. However, in Hepa-1, 3MC and BaP (but not TCDD) caused recruitment of p53 protein to a p53 response element in the 5'-flanking region of the Fmo3 gene. We tested the possibility that FMO3 induction in Hepa-1 cells might be mediated by Nrf2/anti-oxidant response pathways, but agents known to activate Nrf2 or to induce oxidative stress did not affect FMO3 mRNA levels. The protein synthesis inhibitor, cycloheximide (which causes 'superinduction' of CYP1A1 mRNA in TCDD-treated cells), by itself caused dramatic upregulation (> 300-fold) of FMO3 mRNA in Hepa-1 suggesting that cycloheximide prevents synthesis of a labile protein that suppresses FMO3 expression. Although FMO3 mRNA is highly induced by 3MC or TCDD in mouse liver and in Hepa-1 cells, FMO protein levels and FMO catalytic function showed only modest elevation.

  15. Development of doxorubicin-induced chronic cardiotoxicity in the B6C3F{sub 1} mouse model

    SciTech Connect (OSTI)

    Desai, Varsha G.; Herman, Eugene H.; Moland, Carrie L.; Branham, William S.; Lewis, Sherry M.; Davis, Kelly J.; George, Nysia I.; Lee, Taewon; Kerr, Susan; Fuscoe, James C.

    2013-01-01

    Serum levels of cardiac troponins serve as biomarkers of myocardial injury. However, troponins are released into the serum only after damage to cardiac tissue has occurred. Here, we report development of a mouse model of doxorubicin (DOX)-induced chronic cardiotoxicity to aid in the identification of predictive biomarkers of early events of cardiac tissue injury. Male B6C3F{sub 1} mice were administered intravenous DOX at 3 mg/kg body weight, or an equivalent volume of saline, once a week for 4, 6, 8, 10, 12, and 14 weeks, resulting in cumulative DOX doses of 12, 18, 24, 30, 36, and 42 mg/kg, respectively. Mice were sacrificed a week following the last dose. A significant reduction in body weight gain was observed in mice following exposure to a weekly DOX dose for 1 week and longer compared to saline-treated controls. DOX treatment also resulted in declines in red blood cell count, hemoglobin level, and hematocrit compared to saline-treated controls after the 2nd weekly dose until the 8th and 9th doses, followed by a modest recovery. All DOX-treated mice had significant elevations in cardiac troponin T concentrations in plasma compared to saline-treated controls, indicating cardiac tissue injury. Also, a dose-related increase in the severity of cardiac lesions was seen in mice exposed to 24 mg/kg DOX and higher cumulative doses. Mice treated with cumulative DOX doses of 30 mg/kg and higher showed a significant decline in heart rate, suggesting drug-induced cardiac dysfunction. Altogether, these findings demonstrate the development of DOX-induced chronic cardiotoxicity in B6C3F{sub 1} mice. -- Highlights: ? 24 mg/kg was a cumulative cardiotoxic dose of doxorubicin in male B6C3F{sub 1} mice. ? Doxorubicin-induced hematological toxicity was in association with splenomegaly. ? Doxorubicin induced severe testicular toxicity in B6C3F{sub 1} male mice.

  16. Mono-hydroxy methoxychlor alters levels of key sex steroids and steroidogenic enzymes in cultured mouse antral follicles

    SciTech Connect (OSTI)

    Craig, Zelieann R.; Leslie, Traci C.; Hatfield, Kimberly P.; Gupta, Rupesh K.; Flaws, Jodi A.

    2010-12-01

    Methoxychlor (MXC) is an organochlorine pesticide that reduces fertility in female rodents by decreasing antral follicle numbers and increasing follicular death. MXC is metabolized in the body to mono-hydroxy MXC (mono-OH). Little is known about the effects of mono-OH on the ovary. Thus, this work tested the hypothesis that mono-OH exposure decreases production of 17{beta}-estradiol (E{sub 2}) by cultured mouse antral follicles. Antral follicles were isolated from CD-1 mice (age 35-39 days) and exposed to dimethylsulfoxide (DMSO), or mono-OH (0.1-10 {mu}g/mL) for 96 h. Media and follicles were collected for analysis of sex steroid levels and mRNA expression, respectively. Mono-OH treatment (10 {mu}g/mL) decreased E{sub 2} (DMSO: 3009.72 {+-} 744.99 ng/mL; mono-OH 0.1 {mu}g/mL: 1679.66 {+-} 461.99 ng/mL; 1 {mu}g/mL: 1752.72 {+-} 532.41 ng/mL; 10 {mu}g/mL: 45.89 {+-} 33.83 ng/mL), testosterone (DMSO: 15.43 {+-} 2.86 ng/mL; mono-OH 0.1 {mu}g/mL: 17.17 {+-} 4.71 ng/mL; 1 {mu}g/mL: 13.64 {+-} 3.53 ng/mL; 10 {mu}g/mL: 1.29 {+-} 0.23 ng/mL), androstenedione (DMSO: 1.92 {+-} 0.34 ng/mL; mono-OH 0.1 {mu}g/mL: 1.49 {+-} 0.43 ng/mL; 1 {mu}g/mL: 0.64 {+-} 0.31 ng/mL; 10 {mu}g/mL: 0.12 {+-} 0.06 ng/mL) and progesterone (DMSO: 24.11 {+-} 4.21 ng/mL; mono-OH 0.1 {mu}g/mL: 26.77 {+-} 4.41 ng/mL; 1 {mu}g/mL: 20.90 {+-} 3.75 ng/mL; 10 {mu}g/mL: 9.44 {+-} 2.97 ng/mL) levels. Mono-OH did not alter expression of Star, Hsd3b1, Hsd17b1 and Cyp1b1, but it did reduce levels of Cyp11a1, Cyp17a1 and Cyp19a1 mRNA. Collectively, these data suggest that mono-OH significantly decreases levels of key sex steroid hormones and the expression of enzymes required for steroidogenesis.

  17. Linkage mapping of the aldo-2, Pax-5, Ambp, and D4H9S3E loci on mouse chromosome 4 in the region of homology with human chromosome 9

    SciTech Connect (OSTI)

    Pilz, A.; Abbott, C. ); Fountain, J. ); Peters, J. )

    1993-12-01

    The genes for aldolase-B (ALDOB), the [alpha]-microglobulin/bikunin precursor (AMBP), the paired box gene PAX5, and the anonymous DNA marker D9S3 map to human chromosome 9 (HSA9). The authors have set out to map the mouse homologues of each of these genes. The mouse genes for Pax-5 and Ambp previously been shown to map to MMU4. They have used an interspecific backcross to confirm these localizations and to map the mouse homologues of ALDOB (Aldo-2) and D9S3 (D4H9S3E) to the same chromosome. These genes were mapped with respect to the four anchor loci for MMU4. In addition, the panel of backcross DNAs had previously been typed for [delta]-amino levulinate dehydratase (Lv), orosomucoid-1 (Orm-1), and hexabranchion (Hxb), the human homologues of which map to HSA9q. The recombination distances [+-] the standard error between each pair of loci are D4Nds4-1.6[+-]1.1-D4H9S3E-4.0[+-]1.7 (Galt) 0.8[+-]0.8-Pax-5-4.8[+-]1.9-Aldo-2-6.3[+-]2.2-(Lv, Orm-1, Ambp)-1.6[+-]1.1-Hxb-4.0[+-]1.7-Tyrp-1-4.8[+-]1.9-Ifa. The data from this study have extended the known region of conserved synteny between human chromosome 9 and mouse chromosome 4. 17 refs., 2 figs.

  18. Arsenic augments the uptake of oxidized LDL by upregulating the expression of lectin-like oxidized LDL receptor in mouse aortic endothelial cells

    SciTech Connect (OSTI)

    Hossain, Ekhtear; Ota, Akinobu; Karnan, Sivasundaram; Damdindorj, Lkhagvasuren; Takahashi, Miyuki; Konishi, Yuko; Konishi, Hiroyuki; Hosokawa, Yoshitaka

    2013-12-15

    Although chronic arsenic exposure is a well-known risk factor for cardiovascular diseases, including atherosclerosis, the molecular mechanism underlying arsenic-induced atherosclerosis remains obscure. Therefore, this study aimed to elucidate this molecular mechanism. We examined changes in the mRNA level of the lectin-like oxidized LDL (oxLDL) receptor (LOX-1) in a mouse aortic endothelial cell line, END-D, after sodium arsenite (SA) treatment. SA treatment significantly upregulated LOX-1 mRNA expression; this finding was also verified at the protein expression level. Flow cytometry and fluorescence microscopy analyses showed that the cellular uptake of fluorescence (Dil)-labeled oxLDL was significantly augmented with SA treatment. In addition, an anti-LOX-1 antibody completely abrogated the augmented uptake of Dil-oxLDL. We observed that SA increased the levels of the phosphorylated forms of nuclear factor of kappa light polypeptide gene enhancer in B cells (NF-κB)/p65. SA-induced upregulation of LOX-1 protein expression was clearly prevented by treatment with an antioxidant, N-acetylcysteine (NAC), or an NF-κB inhibitor, caffeic acid phenethylester (CAPE). Furthermore, SA-augmented uptake of Dil-oxLDL was also prevented by treatment with NAC or CAPE. Taken together, our results indicate that arsenic upregulates LOX-1 expression through the reactive oxygen species-mediated NF-κB signaling pathway, followed by augmented cellular oxLDL uptake, thus highlighting a critical role of the aberrant LOX-1 signaling pathway in the pathogenesis of arsenic-induced atherosclerosis. - Highlights: • Sodium arsenite (SA) increases LOX-1 expression in mouse aortic endothelial cells. • SA enhances cellular uptake of oxidized LDL in dose-dependent manner. • SA-induced ROS generation enhances phosphorylation of NF-κB. • SA upregulates LOX-1 expression through ROS-activated NF-κB signaling pathway.

  19. BTG/Tob family members Tob1 and Tob2 inhibit proliferation of mouse embryonic stem cells via Id3 mRNA degradation

    SciTech Connect (OSTI)

    Chen, Yuanfan; Wang, Chenchen; Wu, Jenny; Li, Lingsong

    2015-07-03

    The mammalian BTG/Tob family is a group of proteins with anti-proliferative ability, and there are six members including BTG1, BTG2/PC3/Tis21, BTG3/ANA, BTG4/PC3B, Tob1/Tob and Tob2. Among them, Tob subfamily members, specifically Tob1/Tob and Tob2, have the most extensive C-terminal regions. As previously reported, overexpression of BTG/Tob proteins is associated with the inhibition of G1 to S-phase cell cycle progression and decreased cell proliferation in a variety of cell types. Tob subfamily proteins have similar anti-proliferative effects on cell cycle progression in cultured tumor cells. An important unresolved question is whether or not they have function in rapidly proliferating cells, such as embryonic stem cells (ESCs). Tob1 and Tob2 were expressed ubiquitously in mouse ESCs (mESCs), suggesting a possible role in early embryonic development and mESCs. To address the above question and explore the possible functions of the Tob subfamily in ESCs, we established ESCs from different genotypic knockout inner cell mass (ICM). We found that Tob1{sup −/−}, Tob2{sup −/−}, and Tob1/2 double knockout (DKO, Tob1{sup −/−} & Tob2{sup −/−}) ESCs grew faster than wild type (WT) ESCs without losing pluripotency, and we provide a possible mechanistic explanation for these observations: Tob1 and Tob2 inhibit the cell cycle via degradation of Id3 mRNA, which is a set of directly targeted genes of BMP4 signaling in mESCs that play critical roles in the maintenance of ESC properties. Together, our data suggest that BTG/Tob family protein Tob1 and Tob2 regulation cell proliferation does not compromise the basic properties of mESCs. - Highlights: • We established mouse Tob1/2 double knockout embryonic stem cells. • Tob1 and Tob2 inhibit the proliferation of ESCs without effect on pluripotency. • Tob1 and Tob2 involved in the degradation of Id3 in mESCs.

  20. Bisphenol A down-regulates rate-limiting Cyp11a1 to acutely inhibit steroidogenesis in cultured mouse antral follicles

    SciTech Connect (OSTI)

    Peretz, Jackye; Flaws, Jodi A.

    2013-09-01

    Bisphenol A (BPA) is the backbone of polycarbonate plastic products and the epoxy resin lining of aluminum cans. Previous studies have shown that exposure to BPA decreases sex steroid hormone production in mouse antral follicles. The current study tests the hypothesis that BPA first decreases the expression levels of the steroidogenic enzyme cytochrome P450 side-chain cleavage (Cyp11a1) and steroidogenic acute regulatory protein (StAR) in mouse antral follicles, leading to a decrease in sex steroid hormone production in vitro. Further, the current study tests the hypothesis that these effects are acute and reversible after removal of BPA. Exposure to BPA (10 μg/mL and 100 μg/mL) significantly decreased expression of Cyp11a1 and StAR beginning at 18 h and 72 h, respectively, compared to controls. Exposure to BPA (10 μg/mL and 100 μg/mL) significantly decreased progesterone levels beginning at 24 h and decreased androstenedione, testosterone, and estradiol levels at 72 h and 96 h compared to controls. Further, after removing BPA from the culture media at 20 h, expression of Cyp11a1 and progesterone levels were restored to control levels by 48 h and 72 h, respectively. Additionally, expression of StAR and levels of androstenedione, testosterone, and estradiol never decreased compared to controls. These data suggest that BPA acutely decreases expression of Cyp11a1 as early as 18 h and this reduction in Cyp11a1 may lead to a decrease in progesterone production by 24 h, followed by a decrease in androstenedione, testosterone, and estradiol production and expression of StAR at 72 h. Therefore, BPA exposure likely targets Cyp11a1 and steroidogenesis, but these effects are reversible with removal of BPA exposure. - Highlights: • BPA may target Cyp11a1 to inhibit steroidogenesis in antral follicles. • BPA may decrease the expression of Cyp11a1 prior to inhibiting steroidogenesis. • The adverse effects of BPA on steroidogenesis in antral follicles are reversible.

  1. Pregnenolone co-treatment partially restores steroidogenesis, but does not prevent growth inhibition and increased atresia in mouse ovarian antral follicles treated with mono-hydroxy methoxychlor

    SciTech Connect (OSTI)

    Craig, Zelieann R. Hannon, Patrick R. Flaws, Jodi A.

    2013-11-01

    Mono-hydroxy methoxychlor (mono-OH MXC) is a metabolite of the pesticide, methoxychlor (MXC). Although MXC is known to decrease antral follicle numbers, and increase follicle death in rodents, not much is known about the ovarian effects of mono-OH MXC. Previous studies indicate that mono-OH MXC inhibits mouse antral follicle growth, increases follicle death, and inhibits steroidogenesis in vitro. Further, previous studies indicate that CYP11A1 expression and production of progesterone (P{sub 4}) may be the early targets of mono-OH MXC in the steroidogenic pathway. Thus, this study tested whether supplementing pregnenolone, the precursor of progesterone and the substrate for HSD3B, would prevent decreased steroidogenesis, inhibited follicle growth, and increased follicle atresia in mono-OH MXC-treated follicles. Mouse antral follicles were exposed to vehicle (dimethylsulfoxide), mono-OH MXC (10 μg/mL), pregnenolone (1 μg/mL), or mono-OH MXC and pregnenolone together for 96 h. Levels of P{sub 4}, androstenedione (A), testosterone (T), estrone (E{sub 1}), and 17β-estradiol (E{sub 2}) in media were determined, and follicles were processed for histological evaluation of atresia. Pregnenolone treatment alone stimulated production of all steroid hormones except E{sub 2}. Mono-OH MXC-treated follicles had decreased sex steroids, but when given pregnenolone, produced levels of P{sub 4}, A, T, and E{sub 1} that were comparable to those in vehicle-treated follicles. Pregnenolone treatment did not prevent growth inhibition and increased atresia in mono-OH MXC-treated follicles. Collectively, these data support the idea that the most upstream effect of mono-OH MXC on steroidogenesis is by reducing the availability of pregnenolone, and that adding pregnenolone may not be sufficient to prevent inhibited follicle growth and survival. - Highlights: • Mono-OH MXC inhibited antral follicle steroidogenesis, growth, and survival. • Pregnenolone partially restored steroidogenesis in mono-OH MXC-treated follicles. • Pregnenolone did not prevent mono-OH MXC-induced inhibition of growth and survival.

  2. Metabolite signatures in hydrophilic extracts of mouse lungs exposed to cigarette smoke revealed by 1H NMR metabolomics investigation

    SciTech Connect (OSTI)

    Hu, Jian Z.; Wang, Xuan; Feng, Ju; Webb-Robertson, Bobbie-Jo M.; Waters, Katrina M.; Tilton, Susan C.; Pounds, Joel G.; Corley, Richard A.; Liu, Maili; Hu, Mary Y.

    2015-05-12

    Herein, 1H-NMR metabolomics are carried out to evaluate the changes of metabolites in lungs of mice exposed to cigarette smoke. It is found that the concentrations of adenosine derivatives (i.e. ATP, ADP and AMP), inosine and uridine are significantly fluctuated in the lungs of mice exposed to cigarette smoke compared with those of controls regardless the mouse is obese or regular weight. The decreased ATP, ADP, AMP and elevated inosine predict that the deaminases in charge of adenosine derivatives to inosine derivatives conversion are altered in lungs of mice exposed to cigarette smoke. Transcriptional analysis reveals that the concentrations of adenosine monophosphate deaminase and adenosine deaminase are different in the lungs of mice exposed to cigarette smoke, confirming the prediction from metabolomics studies. We also found, for the first time, that the ratio of glycerophosphocholine (GPC) to phosphocholine (PC) is significantly increased in the lungs of obese mice compared with regular weight mice. The ratio of GPC/PC is further elevated in the lungs of obese group by cigarette smoke exposure. Since GPC/PC ratio is a known biomarker for cancer, these results may suggest that obese group is more susceptible to lung cancer when exposed to cigarette smoke.

  3. The combination of glutamate receptor antagonist MK-801 with tamoxifen and its active metabolites potentiates their antiproliferative activity in mouse melanoma K1735-M2 cells

    SciTech Connect (OSTI)

    Ribeiro, Mariana P.C.; Nunes-Correia, Isabel; Santos, Armanda E.; Custdio, Jos B.A.

    2014-02-15

    Recent reports suggest that N-methyl-D-aspartate receptor (NMDAR) blockade by MK-801 decreases tumor growth. Thus, we investigated whether other ionotropic glutamate receptor (iGluR) antagonists were also able to modulate the proliferation of melanoma cells. On the other hand, the antiestrogen tamoxifen (TAM) decreases the proliferation of melanoma cells, and is included in combined therapies for melanoma. As the efficacy of TAM is limited by its metabolism, we investigated the effects of the NMDAR antagonist MK-801 in combination with TAM and its active metabolites, 4-hydroxytamoxifen (OHTAM) and endoxifen (EDX). The NMDAR blockers MK-801 and memantine decreased mouse melanoma K1735-M2 cell proliferation. In contrast, the NMDAR competitive antagonist APV and the AMPA and kainate receptor antagonist NBQX did not affect cell proliferation, suggesting that among the iGluR antagonists only the NMDAR channel blockers inhibit melanoma cell proliferation. The combination of antiestrogens with MK-801 potentiated their individual effects on cell biomass due to diminished cell proliferation, since it decreased the cell number and DNA synthesis without increasing cell death. Importantly, TAM metabolites combined with MK-801 promoted cell cycle arrest in G1. Therefore, the data obtained suggest that the activity of MK-801 and antiestrogens in K1735-M2 cells is greatly enhanced when used in combination. - Highlights: MK-801 and memantine decrease melanoma cell proliferation. The combination of MK-801 with antiestrogens inhibits melanoma cell proliferation. These combinations greatly enhance the effects of the compounds individually. MK-801 combined with tamoxifen active metabolites induces cell cycle arrest in G1. The combination of MK-801 and antiestrogens is an innovative strategy for melanoma.

  4. In-111 chelate conjugates of human transferrin (HTr) and mouse monoclonal anti human transferrin receptor antibody (. cap alpha. HTrR MoAb) for tumor imaging

    SciTech Connect (OSTI)

    Goodwin, D.A.; Meares, C.F.; Diamanti, C.I.; McCall, M.; McTigue, M.; Torti, F.; Martin, B.

    1984-01-01

    At least one of the major pathways of uptake of the commonly used tumor scanning agent Ga-67 is via the transferrin receptor. This suggested the use of stably radio-labeled HTr, and ..cap alpha..HTrR MoAb for tumor imaging in humans. HTr and mouse ..cap alpha..HTrR MoAb were alkylated with 1-(parabromacetamidobenzyl)-EDTA. The mM Alkylproteins, approx. =1 chelate/molecule were labeled with 1-3 mCi In-111 citrate pH/sub 5/ (Sp Act approx. = 100-300 Ci/m mole). Images were made 24 hours after 1 mCi IV and in some patients blood levels, urine excretion and digitized whole body scans were obtained at 1, 24,48 and 96 hours post injection. Ten patients with biopsy proven prostate cancer were studied with In-111 HTr, and four with In-111 ..cap alpha.. HTrR MoAb; all had positive mets on bone scan. In-111 HTr persisted in the circulation with a T1/2 of approx. = four days, approx. = 5%/day being excreted in the urine, to a total of approx. = 60% in 21 days. Nine of ten scans were false negative due to the high blood background. In-111 ..cap alpha..HTrR disappeared rapidly from the blood; with most in the bone marrow at 24 hours. ROI analysis of three patients showed whole body 94% at 24 hours, 89% at 48 hours, and 82% at 96 hours (T1/2 = 10.7 days); liver 19% at 1 hour, 25% at 24 hours, and 21% at 96 hours; spleen 3% at 1 hour, 8% at 24 hours, 7.3% at 48 hours, and 3% at 96 hours. The high bone marrow background allowed only a few of the bone mets seen as bone scan to be visualized. Other tumor types not located in bone may be more easily seen.

  5. Evaluation of Aroclor 1260 exposure in a mouse model of diet-induced obesity and non-alcoholic fatty liver disease

    SciTech Connect (OSTI)

    Wahlang, Banrida; Song, Ming; Beier, Juliane I.; Cameron Falkner, K.; Al-Eryani, Laila; Clair, Heather B.; Prough, Russell A.; Osborne, Tanasa S.; Malarkey, David E.; Christopher States, J.; Cave, Matthew C.

    2014-09-15

    Polychlorinated biphenyls (PCBs) are persistent organic pollutants associated with non-alcoholic fatty liver disease (NAFLD) in epidemiologic studies. The purpose of this study was to evaluate the hepatic effects of a PCB mixture, Aroclor 1260, whose composition mimics human bioaccumulation patterns, in a mouse model of diet-induced obesity (DIO). Male C57Bl/6J mice were fed control diet or 42% high fat diet (HFD) and exposed to Aroclor 1260 (20 mg/kg or 200 mg/kg in corn oil) for 12 weeks. A glucose tolerance test was performed; plasma/tissues were obtained at necropsy for measurements of adipocytokine levels, histology, and gene expression. Aroclor 1260 exposure was associated with decreased body fat in HFD-fed mice but had no effect on blood glucose/lipid levels. Paradoxically, Aroclor 1260 + HFD co-exposed mice demonstrated increased hepatic inflammatory foci at both doses while the degree of steatosis did not change. Serum cytokines, ALT levels and hepatic expression of IL-6 and TNFα were increased only at 20 mg/kg, suggesting an inhibition of pro-inflammatory cytokine production at the 200 mg/kg exposure. Aroclor 1260 induced hepatic expression of cytochrome P450s including Cyp3a11 (Pregnane-Xenobiotic Receptor target) and Cyp2b10 (constitutive androstane receptor target) but Cyp2b10 inducibility was diminished with HFD-feeding. Cyp1a2 (aryl hydrocarbon Receptor target) was induced only at 200 mg/kg. In summary, Aroclor 1260 worsened hepatic and systemic inflammation in DIO. The results indicated a bimodal response of PCB-diet interactions in the context of inflammation which could potentially be explained by xenobiotic receptor activation. Thus, PCB exposure may be a relevant “second hit” in the transformation of steatosis to steatohepatitis. - Highlights: • Aroclor 1260 exposure decreased adiposity in mice fed with high fat diet • Aroclor 1260 exposure induced steatohepatitis in diet-induced obese mice • Aroclor 1260 (20 and 200 mg/kg) induced Cyp2b10 and Cyp3a11 (CAR/PXR target genes) • Aroclor 1260 at 200 mg/kg induced Cyp1a2 (AhR target gene)

  6. Sodium arsenite represses the expression of myogenin in C2C12 mouse myoblast cells through histone modifications and altered expression of Ezh2, Glp, and Igf-1

    SciTech Connect (OSTI)

    Hong, Gia-Ming; Present address: The University of Chicago, Section of Hematology Bain, Lisa J.

    2012-05-01

    Arsenic is a toxicant commonly found in water systems and chronic exposure can result in adverse developmental effects including increased neonatal death, stillbirths, and miscarriages, low birth weight, and altered locomotor activity. Previous studies indicate that 20 nM sodium arsenite exposure to C2C12 mouse myocyte cells delayed myoblast differentiation due to reduced myogenin expression, the transcription factor that differentiates myoblasts into myotubes. In this study, several mechanisms by which arsenic could alter myogenin expression were examined. Exposing differentiating C2C12 cells to 20 nM arsenic increased H3K9 dimethylation (H3K9me2) and H3K9 trimethylation (H3K9me3) by 3-fold near the transcription start site of myogenin, which is indicative of increased repressive marks, and reduced H3K9 acetylation (H3K9Ac) by 0.5-fold, indicative of reduced permissive marks. Protein expression of Glp or Ehmt1, a H3-K9 methyltransferase, was also increased by 1.6-fold in arsenic-exposed cells. In addition to the altered histone remodeling status on the myogenin promoter, protein and mRNA levels of Igf-1, a myogenic growth factor, were significantly repressed by arsenic exposure. Moreover, a 2-fold induction of Ezh2 expression, and an increased recruitment of Ezh2 (3.3-fold) and Dnmt3a (? 2-fold) to the myogenin promoter at the transcription start site (? 40 to + 42), were detected in the arsenic-treated cells. Together, we conclude that the repressed myogenin expression in arsenic-exposed C2C12 cells was likely due to a combination of reduced expression of Igf-1, enhanced nuclear expression and promoter recruitment of Ezh2, and altered histone remodeling status on myogenin promoter (? 40 to + 42). -- Highlights: ? Igf-1 expression is decreased in C2C12 cells after 20 nM arsenite exposure. ? Arsenic exposure alters histone remodeling on the myogenin promoter. ? Glp expression, a H3K9 methyltransferase, was increased in arsenic-exposed cells. ? Ezh2 and Dnmt3a localization to the myogenin promoter is induced by arsenic.

  7. 2,3,7,8-Tetrachlorodibenzo-p-dioxin treatment alters eicosanoid levels in several organs of the mouse in an aryl hydrocarbon receptor-dependent fashion

    SciTech Connect (OSTI)

    Bui, Peter; Solaimani, Parrisa [Molecular Toxicology Program, University of California, Los Angeles, California 90095 (United States) [Molecular Toxicology Program, University of California, Los Angeles, California 90095 (United States); Dept of Pathology and Laboratory Medicine, University of California, Los Angeles, California 90095 (United States); Jonsson Comprehensive Cancer Center, University of California, Los Angeles, California 90095 (United States); Wu, Xiaomeng [Dept of Pathology and Laboratory Medicine, University of California, Los Angeles, California 90095 (United States) [Dept of Pathology and Laboratory Medicine, University of California, Los Angeles, California 90095 (United States); Jonsson Comprehensive Cancer Center, University of California, Los Angeles, California 90095 (United States); Hankinson, Oliver, E-mail: ohank@mednet.ucla.edu [Molecular Toxicology Program, University of California, Los Angeles, California 90095 (United States) [Molecular Toxicology Program, University of California, Los Angeles, California 90095 (United States); Dept of Pathology and Laboratory Medicine, University of California, Los Angeles, California 90095 (United States); Jonsson Comprehensive Cancer Center, University of California, Los Angeles, California 90095 (United States); Molecular Biology Institute, University of California, Los Angeles, California 90095 (United States)

    2012-03-01

    2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) adversely affects many mammalian organs and tissues. These effects are mediated by the aryl hydrocarbon receptor (AHR). CYP1A1, CYP1A2 and CYP1B1 are upregulated by the liganded AHR. These (and other) cytochromes P450 can metabolize arachidonic acid into a variety of bioactive eicosanoids. Towards investigating a potential role of eicosanoids in TCDD toxicity, arachidonic acid, two other unsaturated long-chain fatty acids, and up to twenty-five eicosanoids were measured in five organs/tissues of male and female wild-type and Ahr null mice treated or untreated with TCDD. TCDD generally increased the levels of the four dihydroxyeicosatrienoic acids (DHETs) and (where measured) 5,6-epoxyeicosatrienoic acid and 18-, 19- and 20-hydroxyeicosatrienoic acids (HETEs) in the serum, liver, spleen and lungs, but not the heart, of both sexes, and increased the levels in the serum, liver and spleen of several metabolites that are usually considered products of lipoxygenase activity, but which may also be generated by cytochromes P450. TCDD also increased the levels of the esterified forms of these eicosanoids in the liver in parallel with the corresponding free forms. The levels of prostanoids were generally not affected by TCDD. The above changes did not occur in Ahr null mice, and are therefore mediated by the AHR. TCDD increased the mRNA levels of Cyp1a1, Cyp1a2, Cyp1b1 and the Pla2g12a form of phospholipase A{sub 2} to varying degrees in the different organs, and these increases correlated with some but not all the changes in eicosanoids levels in the organs, suggesting that other enzymes may also be involved. -- Highlights: ? TCDD treatment increases the levels of many eicosanoids in several mouse organs. ? Products of both the cytochrome P450 and classical lipoxygenase pathways are increased. ? These increases are dependent on the aryl hydrocarbon receptor. ? Cyp1a1, Cyp1a2 and Cyp1b1 appear to be responsible for much but not all of the increases.

  8. 9. international mouse genome conference

    SciTech Connect (OSTI)

    1995-12-31

    This conference was held November 12--16, 1995 in Ann Arbor, Michigan. The purpose of this conference was to provide a multidisciplinary forum for exchange of state-of-the-art information on genetic mapping in mice. This report contains abstracts of presentations, focusing on the following areas: mutation identification; comparative mapping; informatics and complex traits; mutagenesis; gene identification and new technology; and genetic and physical mapping.

  9. 2,3,7,8-Tetrachlorodibenzo-p-dioxin activates the aryl hydrocarbon receptor and alters sex steroid hormone secretion without affecting growth of mouse antral follicles in vitro

    SciTech Connect (OSTI)

    Karman, Bethany N. Basavarajappa, Mallikarjuna S. Craig, Zelieann R. Flaws, Jodi A.

    2012-05-15

    The persistent environmental contaminant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is an ovarian toxicant. These studies were designed to characterize the actions of TCDD on steroidogenesis and growth of intact mouse antral follicles in vitro. Specifically, these studies tested the hypothesis that TCDD exposure leads to decreased sex hormone production/secretion by antral follicles as well as decreased growth of antral follicles in vitro. Since TCDD acts through binding to the aryl hydrocarbon receptor (AHR), and the AHR has been identified as an important factor in ovarian function, we also conducted experiments to confirm the presence and activation of the AHR in our tissue culture system. To do so, we exposed mouse antral follicles for 96 h to a series of TCDD doses previously shown to have effects on ovarian tissues and cells in culture, which also encompass environmentally relevant and pharmacological exposures (0.1–100 nM), to determine a dose response for TCDD in our culture system for growth, hormone production, and expression of the Ahr and Cyp1b1. The results indicate that TCDD decreases progesterone, androstenedione, testosterone, and estradiol levels in a non-monotonic dose response manner without altering growth of antral follicles. The addition of pregnenolone substrate (10 μM) restores hormone levels to control levels. Additionally, Cyp1b1 levels were increased by 3–4 fold regardless of the dose of TCDD exposure, evidence of AHR activation. Overall, these data indicate that TCDD may act prior to pregnenolone formation and through AHR transcriptional control of Cyp1b1, leading to decreased hormone levels without affecting growth of antral follicles. -- Highlights: ►TCDD disrupts sex steroid hormone levels, but not growth of antral follicles. ►Pregnenolone co-treatment by-passes TCDD-induced steroid hormone disruption. ►TCDD affects steroid hormone levels through an AHR pathway in antral follicles.

  10. Notch-modifying xylosyltransferase structures support an SNi-like retaining mechanism

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Yu, Hongjun; Li, Huilin; Takeuchi, Megumi; LeBarron, Jamie; Kantharia, Joshua; London, Erwin; Bakker, Hans; Haltiwanger, Robert S.; Takeuchi, Hideyuki

    2015-09-28

    A major question remaining in glycobiology is how a glycosyltransferase (GT) that retains the anomeric linkage of a sugar catalyzes the reaction. Xyloside α-1,3-xylosyltransferase (XXYLT1) is a retaining GT that regulates Notch receptor activation by adding xylose to the Notch extracellular domain. Here, using natural acceptor and donor substrates and active Mus musculus XXYLT1, we report a series of crystallographic snapshots along the reaction, including an unprecedented natural and competent Michaelis reaction complex for retaining enzymes. These structures strongly support the SNi-like reaction as the retaining mechanism for XXYLT1. Unexpectedly, the epidermal growth factor–like repeat acceptor substrate undergoes a largemore » conformational change upon binding to the active site, providing a structural basis for substrate specificity. As a result, our improved understanding of this retaining enzyme will accelerate the design of retaining GT inhibitors that can modulate Notch activity in pathological situations in which Notch dysregulation is known to cause cancer or developmental disorders.« less

  11. Notch-modifying xylosyltransferase structures support an SNi-like retaining mechanism

    SciTech Connect (OSTI)

    Yu, Hongjun; Li, Huilin; Takeuchi, Megumi; LeBarron, Jamie; Kantharia, Joshua; London, Erwin; Bakker, Hans; Haltiwanger, Robert S.; Takeuchi, Hideyuki

    2015-09-28

    A major question remaining in glycobiology is how a glycosyltransferase (GT) that retains the anomeric linkage of a sugar catalyzes the reaction. Xyloside ?-1,3-xylosyltransferase (XXYLT1) is a retaining GT that regulates Notch receptor activation by adding xylose to the Notch extracellular domain. Here, using natural acceptor and donor substrates and active Mus musculus XXYLT1, we report a series of crystallographic snapshots along the reaction, including an unprecedented natural and competent Michaelis reaction complex for retaining enzymes. These structures strongly support the SNi-like reaction as the retaining mechanism for XXYLT1. Unexpectedly, the epidermal growth factorlike repeat acceptor substrate undergoes a large conformational change upon binding to the active site, providing a structural basis for substrate specificity. As a result, our improved understanding of this retaining enzyme will accelerate the design of retaining GT inhibitors that can modulate Notch activity in pathological situations in which Notch dysregulation is known to cause cancer or developmental disorders.

  12. Absence of correlation between Sry polymorphisms and XY sex reversal caused by the M.m. domesticus Y chromosome

    SciTech Connect (OSTI)

    Carlisle, C.; Nagamine, C.M. [Vanderbilt Univ., School of Medicine, Nashville, TN (United States)] [Vanderbilt Univ., School of Medicine, Nashville, TN (United States); Winkinig, H.; Weichenhan, D. [Medizinische Universitaet Zu Luebeck (Germany)] [Medizinische Universitaet Zu Luebeck (Germany)

    1996-04-01

    Mus musculus domesticus Y chromosomes (Y{sup DOM} Chrs) vary in their ability to induce testes in the strain C57BL/6J. In severe cases, XY females develop (XY{sup DOM} sex reversal). To identify the molecular basis for the sex reversal, a 2.7-kb region of Sry, the testis-determining gene, was sequenced from Y{sup DOM} Chrs linked to normal testis determination, transient sex reversal, and severe sex reversal. Four mutations were identified. However, no correlation exists between these mutations and severity of XY{sup DOM} sex reversal. RT-PCR identified Sry transcripts in XY{sup DOM} sex-reversed fetal gonads at 11 d.p.c., the age when Sry is hypothesized to function. In addition, no correlation exists between XY{sup DOM} sex reversal and copy numbers of pSx1, a Y-repetitive sequence whose deletion is linked to XY sex reversal. We conclude that SRY protein variants, blockade of Sry transcription, and deletion of pSx1 sequences are not the underlying causes of XY{sup DOM} sex reversal. 63 refs., 6 figs., 6 tabs.

  13. Arsenic and chromium in drinking water promote tumorigenesis in a mouse colitis-associated colorectal cancer model and the potential mechanism is ROS-mediated Wnt/β-catenin signaling pathway

    SciTech Connect (OSTI)

    Wang, Xin; Mandal, Ardhendu K.; Saito, Hiroshi; Pulliam, Joseph F.; Lee, Eun Y.; Ke, Zun-Ji; Lu, Jian; Ding, Songze; Li, Li; Shelton, Brent J.; Tucker, Thomas; Evers, B. Mark; Zhang, Zhuo; Shi, Xianglin

    2012-07-01

    Exposure to carcinogenic metals, such as trivalent arsenic [As(III)] and hexavalent chromium [Cr(VI)], through drinking water is a major global public health problem and is associated with various cancers. However, the mechanism of their carcinogenicity remains unclear. In this study, we used azoxymethane/dextran sodium sulfate (AOM/DSS)-induced mouse colitis-associated colorectal cancer model to investigate their tumorigenesis. Our results demonstrate that exposure to As(III) or Cr(VI), alone or in combination, together with AOM/DSS pretreatment has a promotion effect, increasing the colorectal tumor incidence, multiplicity, size, and grade, as well as cell inflammatory response. Two-dimensional differential gel electrophoresis coupled with mass spectrometry revealed that As(III) or Cr(VI) treatment alone significantly changed the density of proteins. The expression of β-catenin and phospho-GSK was increased by treatment of carcinogenic metals alone. Concomitantly, the expression of NADPH oxidase1 (NOX1) and the level of 8-OHdG were also increased by treatment of carcinogenic metals alone. Antioxidant enzymes, such as superoxide dismutase (SOD) and catalase, were decreased. Similarly, in an in vitro system, exposure of CRL-1807 to carcinogenic metals increased reactive oxygen species (ROS) generation, the expression of β-catenin, phospho-GSK, and NOX1. Inhibition of ROS generation by addition of SOD or catalase inhibited β-catenin expression and activity. Our study provides a new animal model to study the carcinogenicity of As(III) and Cr(VI) and suggests that As(III) and Cr(VI) promote colorectal cancer tumorigenesis, at least partly, through ROS-mediated Wnt/β-catenin signaling pathway. -- Highlights: ► Carcinogenic metals in drinking water promote colorectal tumor formation in vivo. ► Carcinogenic metals induce β-catenin activation in vivo and in vitro. ► ROS generation induced by carcinogenic metals mediated β-catenin activation.

  14. Reduction of metastasis, cell invasion, and adhesion in mouse osteosarcoma by YM529/ONO-5920-induced blockade of the Ras/MEK/ERK and Ras/PI3K/Akt pathway

    SciTech Connect (OSTI)

    Tsubaki, Masanobu; Satou, Takao; Itoh, Tatsuki; Imano, Motohiro; Ogaki, Mitsuhiko; Yanae, Masashi; Depeartment of Pharmacy, Sakai Hospital, Kinki University School of Medicine, Sakai, Osaka 590-0132 ; Nishida, Shozo

    2012-03-15

    Osteosarcoma is one of the most common primary malignant bone tumors in children and adolescents. Some patients continue to have a poor prognosis, because of the metastatic disease. YM529/ONO-5920 is a nitrogen-containing bisphosphonate that has been used for the treatment of osteoporosis. YM529/ONO-5920 has recently been reported to induce apoptosis in various tumors including osteosarcoma. However, the mode of metastasis suppression in osteosarcoma by YM529/ONO-5920 is unclear. In the present study, we investigated whether YM529/ONO-5920 inhibited tumor cell migration, invasion, adhesion, or metastasis in the LM8 mouse osteosarcoma cell line. We found that YM529/ONO-5920 significantly inhibited metastasis, cell migration, invasion, and adhesion at concentrations that did not have antiproliferative effects on LM8 cells. YM529/ONO-5920 also inhibited the mRNA expression and protein activities of matrix metalloproteinases (MMPs). In addition, YM529/ONO-5920 suppressed phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2) and the serine/threonine protein kinase B (Akt) by the inhibition of Ras prenylation. Moreover, U0126, a mitogen-activated protein kinase kinase (MEK) 1/2 inhibitor, and LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, also inhibited LM8 cell migration, invasion, adhesion, and metastasis, as well as the mRNA expression and protein activities of MMP-1, MMP-2, MMP-9, and MT1-MMP. The results indicated that YM529/ONO-5920 suppressed the Ras/MEK/ERK and Ras/PI3K/Akt pathways, thereby inhibiting LM8 cell migration, invasion, adhesion, and metastasis. These findings suggest that YM529/ONO-5920 has potential clinical applications for the treatment of tumor cell metastasis in osteosarcoma. -- Highlights: ? We investigated whether YM529/ONO-5920 inhibited tumor metastasis in osteosarcoma. ? YM529/ONO-5920 inhibited metastasis, cell migration, invasion, and adhesion. ? YM529/ONO-5920 suppressed Ras signalings. ? YM529/ONO-5920 has potential clinical applications for the treatment in osteosarcoma.

  15. Sandia Energy - Sandia Maps Multiple Paths to Cleaner, Low-Temp...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Mark Musculus and his colleagues identified the sources of key pollutants from LTC engines. Understanding how LTC works as a combustion technique may lead to broader use of...

  16. TU-F-12A-01: Quantitative Non-Linear Compartment Modeling of 89Zr- and 124I- Labeled J591 Monoclonal Antibody Kinetics Using Serial Non-Invasive Positron Emission Tomography Imaging in a Pre-Clinical Human Prostate Cancer Mouse Model

    SciTech Connect (OSTI)

    Fung, EK; Cheal, SM; Chalasani, S; Fareedy, SB; Punzalan, B; Humm, JL; Osborne, JR; Larson, SM; Zanzonico, PB; Otto, B; Bander, NH

    2014-06-15

    Purpose: To examine the binding kinetics of human IgG monoclonal antibody J591 which targets prostate-specific membrane antigen (PSMA) in a pre-clinical mouse cancer model using quantitative PET compartmental analysis of two radiolabeled variants. Methods: PSMA is expressed in normal human prostate, and becomes highly upregulated in prostate cancer, making it a promising therapeutic target. Two forms of J591, radiolabeled with either {sup 89}Zr or {sup 124}I, were prepared. {sup 89}Zr is a radiometal that becomes trapped in the cell upon internalization by the antigen-antibody complex, while radioiodine leaves the cell. Mice with prostate cancer xenografts underwent non-invasive serial imaging on a Focus 120 microPET up to 144 hours post-injection of J591. A non-linear compartmental model describing the binding and internalization of antibody in tumor xenograft was developed and applied to the PET-derived time-activity curves. The antibody-antigen association rate constant (ka), total amount of antigen per gram tumor (Ag-total), internalization rate of antibody-antigen complex, and efflux rate of radioisotope from tumor were fitted using the model. The surface-bound and the internalized activity were also estimated. Results: Values for ka, Ag-total, and internalization rate were found to be similar regardless of radiolabel payload used. The efflux rate, however, was ? 9-fold higher for {sup 124}I-J591 than for {sup 89}Zr-J591. Time-dependent surface-bound and internalized radiotracer activity were similar for both radiolabels at early times post-injection, but clearly differed beyond 24 hours. Conclusion: Binding and internalization of J591 to PSMA-expressing tumor xenografts were similar when radiolabeled with either {sup 89}Zr or {sup 124}I payload. The difference in efflux of radioactivity from tumor may be attributable to differential biological fate intracellularly of the radioisotopes. This has great significance for radioimmunotherapy and antibody-drug conjugates. Further exploration using the model will examine binding and radioisotope residence as antibody dose is increased to antigen saturation. The Center for Targeted Radioimmunotherapy and Theranostics, Ludwig Center for Cancer Immunotherapy, Memorial Sloan Kettering Cancer Center (MSK), NIH (R25-CA096945). Technical services provided by the MSK Small-Animal Imaging Core Facility were supported by the NIH (R24-CA83084, P30-CA08748, and P50-CA92629; Zanzonico). NCI, Center to Reduce Cancer Health Disparity (R21 CA153177-03; Osborne)

  17. Metabonomic Profiling of TASTPM Transgenic Alzheimer's Disease Mouse Model

    SciTech Connect (OSTI)

    Hu, Zeping; Browne, Edward R.; Liu, Tao; Angel, Thomas E.; Ho, Paul C.; Chun Yong Chan, Eric

    2012-12-07

    Identification of molecular mechanisms underlying early stage Alzheimer’s disease (AD) is important for the development of new therapies against and diagnosis of AD. In this study, non-targeted metabotyping of TASTPM transgenic AD mice was performed. The metabolic profiles of both brain and plasma of TASTPM mice were characterized using gas chromatography-mass spectrometry and compared to those of wild type C57BL/6J mice. TASTPM mice were metabolically distinct compared to wild type mice (Q28 Y = 0.587 and 0.766 for PLS-DA models derived from brain and plasma, respectively). A number of metabolites were found to be perturbed in TASTPM mice in both brain (D11 fructose, L-valine, L-serine, L-threonine, zymosterol) and plasma (D-glucose, D12 galactose, linoleic acid, arachidonic acid, palmitic acid and D-gluconic acid). In addition, enzyme immunoassay confirmed that selected endogenous steroids were significantly perturbed in brain (androstenedione and 17-OH-progesterone) and plasma (cortisol and testosterone) of TASTPM mice. Ingenuity pathway analysis revealed that perturbations related to amino acid metabolism (brain), steroid biosynthesis (brain), linoleic acid metabolism (plasma) and energy metabolism (plasma) accounted for the differentiation of TASTPM and wild-type

  18. RAPID/Best Practices/Memorandums of Understanding (MOUs) | Open...

    Open Energy Info (EERE)

    Agencies. If necessary, enable (through legislation or otherwise) state agencies to enter into agreements with the agencies of other states to establish consistent technical...

  19. Saving Money and Fuel with a Click of a Mouse

    Broader source: Energy.gov [DOE]

    With so many options, it can be hard to decipher what car is right for you, or if there’s a clear economic benefit in trading up to a new vehicle. Fortunately, the Energy Department offers a number of tools that can help consumers save money and fuel, whether you’re in the market for a new vehicle or trying to make the most of your current one.

  20. The Crystal Structure of Mouse Exo70 Reveals Unique Features...

    Office of Scientific and Technical Information (OSTI)

    OSTI Identifier: 1186909 Resource Type: Journal Article Resource Relation: Journal Name: Journal of Molecular Biology; Journal Volume: 371; Journal Issue: (2) ; 08, 2007 Publisher: ...

  1. Cleaner, More Efficient Diesel Engines

    SciTech Connect (OSTI)

    Musculus, Mark

    2013-08-13

    Mark Musculus, an engine combustion scientist at Sandia National Laboratories, led a study that outlines the science base for auto and engine manufacturers to build the next generation of cleaner, more efficient engines using low-temperature combustion. Here, Musculus discusses the work at Sandia's Combustion Research Facility.

  2. Cleaner, More Efficient Diesel Engines

    ScienceCinema (OSTI)

    Musculus, Mark

    2014-02-26

    Mark Musculus, an engine combustion scientist at Sandia National Laboratories, led a study that outlines the science base for auto and engine manufacturers to build the next generation of cleaner, more efficient engines using low-temperature combustion. Here, Musculus discusses the work at Sandia's Combustion Research Facility.

  3. Heavy-Duty Low-Temperature and Diesel Combustion & Heavy-Duty...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Peer Evaluation PDF icon ace001musculus2011o.pdf More Documents & Publications Heavy-Duty Low-Temperature and Diesel Combustion & Heavy-Duty Combustion Modeling Heavy-Duty ...

  4. Preliminary X-ray crystallographic studies of mouse UPR responsive protein P58(IPK) TPR fragment

    SciTech Connect (OSTI)

    Tao, Jiahui; Wu, Yunkun [Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL 35294 (United States); Ron, David [Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY 10016 (United States); Sha, Bingdong, E-mail: bdsha@uab.edu [Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL 35294 (United States)

    2008-02-01

    To investigate the mechanism by which P58(IPK) functions to promote protein folding within the ER, a P58(IPK) TPR fragment without the C-terminal J-domain has been crystallized. Endoplasmic reticulum (ER) stress induces the unfolded protein response (UPR), which can promote protein folding and misfolded protein degradation and attenuate protein translation and protein translocation into the ER. P58(IPK) has been proposed to function as a molecular chaperone to maintain protein-folding homeostasis in the ER under normal and stressed conditions. P58(IPK) contains nine TPR motifs and a C-terminal J-domain within its primary sequence. To investigate the mechanism by which P58(IPK) functions to promote protein folding within the ER, a P58(IPK) TPR fragment without the C-terminal J-domain was crystallized. The crystals diffract to 2.5 resolution using a synchrotron X-ray source. The crystals belong to space group P2{sub 1}, with unit-cell parameters a = 83.53, b = 92.75, c = 84.32 , ? = 90.00, ? = 119.36, ? = 90.00. There are two P58(IPK) molecules in the asymmetric unit, which corresponds to a solvent content of approximately 60%. Structure determination by MAD methods is under way.

  5. Carbamazepine suppresses calpain-mediated autophagy impairment after ischemia/reperfusion in mouse livers

    SciTech Connect (OSTI)

    Kim, Jae-Sung, E-mail: Jae.Kim@surgery.ufl.edu; Wang, Jin-Hee, E-mail: jin-hee.wang@surgery.ufl.edu; Biel, Thomas G., E-mail: Thomas.Biel@surgery.ufl.edu; Kim, Do-Sung, E-mail: do-sung.kim@surgery.med.ufl.edu; Flores-Toro, Joseph A., E-mail: Joseph.Flores-Toro@surgery.ufl.edu; Vijayvargiya, Richa, E-mail: rvijayvargiya@ufl.edu; Zendejas, Ivan, E-mail: ivan.zendejas@surgery.ufl.edu; Behrns, Kevin E., E-mail: Kevin.Behrns@surgery.ufl.edu

    2013-12-15

    Onset of the mitochondrial permeability transition (MPT) plays a causative role in ischemia/reperfusion (I/R) injury. Current therapeutic strategies for reducing reperfusion injury remain disappointing. Autophagy is a lysosome-mediated, catabolic process that timely eliminates abnormal or damaged cellular constituents and organelles such as dysfunctional mitochondria. I/R induces calcium overloading and calpain activation, leading to degradation of key autophagy-related proteins (Atg). Carbamazepine (CBZ), an FDA-approved anticonvulsant drug, has recently been reported to increase autophagy. We investigated the effects of CBZ on hepatic I/R injury. Hepatocytes and livers from male C57BL/6 mice were subjected to simulated in vitro, as well as in vivo I/R, respectively. Cell death, intracellular calcium, calpain activity, changes in autophagy-related proteins (Atg), autophagic flux, MPT and mitochondrial membrane potential after I/R were analyzed in the presence and absence of 20 ?M CBZ. CBZ significantly increased hepatocyte viability after reperfusion. Confocal microscopy revealed that CBZ prevented calcium overloading, the onset of the MPT and mitochondrial depolarization. Immunoblotting and fluorometric analysis showed that CBZ blocked calpain activation, depletion of Atg7 and Beclin-1 and loss of autophagic flux after reperfusion. Intravital multiphoton imaging of anesthetized mice demonstrated that CBZ substantially reversed autophagic defects and mitochondrial dysfunction after I/R in vivo. In conclusion, CBZ prevents calcium overloading and calpain activation, which, in turn, suppresses Atg7 and Beclin-1 depletion, defective autophagy, onset of the MPT and cell death after I/R. - Highlights: A mechanism of carbamazepine (CBZ)-induced cytoprotection in livers is proposed. Impaired autophagy is a key event contributing to lethal reperfusion injury. The importance of autophagy is extended and confirmed in an in vivo model. CBZ is a potential agent to improve liver function after liver surgery.

  6. Radioprotective effect of combinations of WR-2721 and mercaptopropionylglycine on mouse bone marrow chromosomes

    SciTech Connect (OSTI)

    Uma Devi, P.; Prasanna, P.G. )

    1990-11-01

    The radioprotective and toxic effects of low to moderate doses of S-2-(3-aminopropylamino)ethyl phosphorothioic acid (WR-2721) and its combination with mercaptopropionylglycine (MPG) on the chromosomes of the bone marrow cells of Swiss albino mice were studied at 24 h and 14 days postirradiation. Significant protection against radiation-induced chromosome aberrations was observed with 50 mg/kg WR-2721. The protection increased with the dose of the drug administered, and the degree of protection per unit dose increment was more pronounced at lower than at higher doses. A combination of WR-2721 and MPG given before exposure resulted in a significantly greater number of normal metaphases at 24 h postirradiation compared to the respective single-drug treatment groups. On Day 14 postirradiation, when the presence of WR-2721 resulted in an increase in the frequency of aberrant cells, combination with MPG helped to reduce this value markedly, especially at WR-2721 doses below 200 mg/kg. On the basis of these results it is suggested that 150 mg/kg WR-2721 may be considered an optimum dose for combination with MPG for protection of chromosomes of bone marrow cells when repeated drug administrations are not needed. Changes in the level of glutathione (GSH) in the blood were studied at different times following the administration of 150 mg/kg WR-2721 and its combination with MPG before sham irradiation or exposure to 4.5 Gy 60Co gamma rays. The results showed that WR-2721 elevated blood GSH levels significantly above normal values by the time radiation was delivered, while MPG did not. Glutathione appears to have an important role in the action of WR-2721, while protection by MPG may not be mediated through GSH. Injection of MPG after WR-2721 helps to maintain the higher GSH level for a longer duration compared to treatment with WR-2721 alone.

  7. Scanning Tranmission X-ray Microscopic Analysis of Purifed Melanosomes of the Mouse Iris

    SciTech Connect (OSTI)

    Anderson,M.; Haraszti, T.; Peterson, G.; Wirick, S.; Jacobsen, C.; John, S.; Grunze, M.

    2006-01-01

    Melanosomes are specialized intracellular membrane bound organelles that produce and store melanin pigment. The composition of melanin and distribution of melanosomes determine the color of many mammalian tissues, including the hair, skin, and iris. However, the presence of melanosomes within a tissue carries potentially detrimental risks related to the cytotoxic indole-quinone intermediates produced during melanin synthesis. In order to study melanosomal molecules, including melanin and melanin-related intermediates, we have refined methods allowing spectromicroscopic analysis of purified melanosomes using scanning transmission X-ray microscopy. Here, we present for the first time absorption data for melanosomes at the carbon absorption edge ranging from 284 to 290 eV. High-resolution images of melanosomes at discrete energies demonstrate that fully melanized mature melanosomes are internally non-homogeneous, suggesting the presence of an organized internal sub-structure. Spectra of purified melanosomes are complex, partially described by a predominating absorption band at 288.4 eV with additional contributions from several minor bands. Differences in these spectra were detectable between samples from two strains of inbred mice known to harbor genetically determined melanosomal differences, DBA/2J and C57BL/6J, and are likely to represent signatures arising from biologically relevant and tractable phenomena.

  8. Microsoft Word - FINAL 2010_SPFPA_CBA With MOUs.doc

    National Nuclear Security Administration (NNSA)

    AGREEMENT between Honeywell Federal Manufacturing and Technologies, LLC and The International Union, Security, Police and Fire Professionals of America and its Amalgamated Local No. 251 Effective August 23, 2010 through August 21, 2016 2 TABLE OF CONTENTS Page 1. AGREEMENT ................................................................ 1 2. INTENT ........................................................................ 1 3. RECOGNITION

  9. Mauritius: Energy Resources | Open Energy Information

    Open Energy Info (EERE)

    "","visitedicon":"" Country Profile Name Mauritius Population 1,236,817 GDP 14 Energy Consumption 0.06 Quadrillion Btu 2-letter ISO code MU 3-letter ISO code MUS Numeric ISO...

  10. The role of acetate in alcohol-induced alterations of uterine glucose metabolism in the mouse during pregnancy

    SciTech Connect (OSTI)

    Simm, B. ); Murdoch, R.N. )

    1990-01-01

    The acute exposure of mice to ethanol during post-implantation pregnancy has been reported to cause alterations in the levels of several glycolytic intermediates in the uterus, suggesting a possible indirect mechanism of alcohol embryo-toxicity. The present study was undertaken to assess whether the ethanol metabolite, acetate is implicated in this phenomenon. Blood and uterine alcohol concentrations in day 9 - pregnant Quackenbush Swiss mice were maximal 15 minutes after the intraperitoneal injection of ethanol, and fell to almost negligible levels 6 hours later. In response to this treatment, the levels of blood and uterine acetate increased, liver glycogen decreased, plasma glucose increased, and uterine glucose, glucose-6-phosphate (G-6-P), fructose-6-phosphate (F-6-P), and citrate increased. When acetate was administered to pregnant mice in amounts approximating those generated by exposure to alcohol, the levels of uterine F-6-P and citrate increased while other metabolic parameters remained unaffected. The administration of 4-methylpyrazole to mice subsequently treated with alcohol produced conditions of alcohol exposure in the absence of ethanol-derived acetate and depressed the ethanol-induced rise in uterine G-6-P and citrate.

  11. Regulation Of Nf=kb And Mnsod In Low Dose Radiation Induced Adaptive Protection Of Mouse And Human Skin Cells

    SciTech Connect (OSTI)

    Jian Li

    2012-11-07

    A sampling of publications resulting from this grant is provided. One is on the subject of NF-κB-Mediated HER2 Overexpression in Radiation-Adaptive Resistance. Another is on NF-κB-mediated adaptive resistance to ionizing radiation.

  12. Ultraviolet germicidal irradiation and its effects on elemental distributions in mouse embryonic fibroblast cells in x-ray fluorescence microanalysis

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Jin, Qiaoling; Vogt, Stefan; Lai, Barry; Chen, Si; Finney, Lydia; Gleber, Sophie-Charlotte; Ward, Jesse; Deng, Junjing; Mak, Rachel; Moonier, Nena; et al

    2015-02-23

    Rapidly-frozen hydrated (cryopreserved) specimens combined with cryo-scanning x-ray fluorescence microscopy provide an ideal approach for investigating elemental distributions in biological cells and tissues. However, because cryopreservation does not deactivate potentially infectious agents associated with Risk Group 2 biological materials, one must be concerned with contamination of expensive and complicated cryogenic x-ray microscopes when working with such materials. We employed ultraviolet germicidal irradiation to decontaminate previously cryopreserved cells under liquid nitrogen, and then investigated its effects on elemental distributions under both frozen hydrated and freeze dried states with x-ray fluorescence microscopy. We show that the contents and distributions of most biologicallymore » important elements remain nearly unchanged when compared with non-ultraviolet-irradiated counterparts, even after multiple cycles of ultraviolet germicidal irradiation and cryogenic x-ray imaging. This provides a potential pathway for rendering Risk Group 2 biological materials safe for handling in multiuser cryogenic x-ray microscopes without affecting the fidelity of the results.« less

  13. Biodiesel versus diesel exposure: Enhanced pulmonary inflammation, oxidative stress, and differential morphological changes in the mouse lung

    SciTech Connect (OSTI)

    Yanamala, Naveena; Birch, M. Eileen; Kisin, Elena; Bugarski, Aleksandar D.

    2013-10-15

    The use of biodiesel (BD) or its blends with petroleum diesel (D) is considered to be a viable approach to reduce occupational and environmental exposures to particulate matter (PM). Due to its lower particulate mass emissions compared to D, use of BD is thought to alleviate adverse health effects. Considering BD fuel is mainly composed of unsaturated fatty acids, we hypothesize that BD exhaust particles could induce pronounced adverse outcomes, due to their ability to readily oxidize. The main objective of this study was to compare the effects of particles generated by engine fueled with neat BD and neat petroleum-based D. Biomarkers of tissue damage and inflammation were significantly elevated in lungs of mice exposed to BD particulates. Additionally, BD particulates caused a significant accumulation of oxidatively modified proteins and an increase in 4-hydroxynonenal. The up-regulation of inflammatory cytokines/chemokines/growth factors was higher in lungs upon BD particulate exposure. Histological evaluation of lung sections indicated presence of lymphocytic infiltrate and impaired clearance with prolonged retention of BD particulate in pigment laden macrophages. Taken together, these results clearly indicate that BD exhaust particles could exert more toxic effects compared to D. - Highlights: Exposure of mice to BDPM caused higher pulmonary toxicity compared to DPM. Oxidative stress and inflammation were higher in BD vs to D exposed mice. Inflammatory lymphocyte infiltrates were seen only in lungs of mice exposed to BD. Ineffective clearance, prolonged PM retention was present only after BD exposure.

  14. Ultraviolet germicidal irradiation and its effects on elemental distributions in mouse embryonic fibroblast cells in x-ray fluorescence microanalysis

    SciTech Connect (OSTI)

    Jin, Qiaoling; Vogt, Stefan; Lai, Barry; Chen, Si; Finney, Lydia; Gleber, Sophie-Charlotte; Ward, Jesse; Deng, Junjing; Mak, Rachel; Moonier, Nena; Jacobsen, Chris; Brody, James P.

    2015-02-23

    Rapidly-frozen hydrated (cryopreserved) specimens combined with cryo-scanning x-ray fluorescence microscopy provide an ideal approach for investigating elemental distributions in biological cells and tissues. However, because cryopreservation does not deactivate potentially infectious agents associated with Risk Group 2 biological materials, one must be concerned with contamination of expensive and complicated cryogenic x-ray microscopes when working with such materials. We employed ultraviolet germicidal irradiation to decontaminate previously cryopreserved cells under liquid nitrogen, and then investigated its effects on elemental distributions under both frozen hydrated and freeze dried states with x-ray fluorescence microscopy. We show that the contents and distributions of most biologically important elements remain nearly unchanged when compared with non-ultraviolet-irradiated counterparts, even after multiple cycles of ultraviolet germicidal irradiation and cryogenic x-ray imaging. This provides a potential pathway for rendering Risk Group 2 biological materials safe for handling in multiuser cryogenic x-ray microscopes without affecting the fidelity of the results.

  15. Focused ultrasound treatment of abscesses induced by methicillin resistant Staphylococcus aureus: Feasibility study in a mouse model

    SciTech Connect (OSTI)

    Rieck, Birgit; Bates, David; Pichardo, Samuel E-mail: lcuriel@lakeheadu.ca; Curiel, Laura E-mail: lcuriel@lakeheadu.ca; Zhang, Kunyan; Escott, Nicholas; Mougenot, Charles

    2014-06-15

    Purpose: To study the therapeutic effect of focused ultrasound on abscesses induced by methicillin-resistantStaphylococcus aureus (MRSA). MRSA is a major nosocomial pathogen where immunocompromised patients are prone to develop infections that are less and less responsive to regular treatments. Because of its capability to induce a rise of temperature at a very precise location, the use of focused ultrasound represents a considerable opportunity for therapy of localized MRSA-related infections. Methods: 50μl of MRSA strain USA400 bacteria suspension at a concentration of 1.32 ± 0.5 × 10{sup 5} colony forming units (cfu)/μl was injected subcutaneously in the left flank of BALB/c mice. An abscess of 6 ± 2 mm in diameter formed after 48 h. A transducer operating at 3 MHz with a focal length of 50 mm and diameter of 32 mm was used to treat the abscess. The focal point was positioned 2 mm under the skin at the abscess center. Forty-eight hours after injection four ultrasound exposures of 9 s each were applied to each abscess under magnetic resonance imaging guidance. Each exposure was followed by a 1 min pause. These parameters were based on preliminary experiments to ensure repetitive accurate heating of the abscess. Real-time estimation of change of temperature was done using water-proton resonance frequency and a communication toolbox (matMRI) developed inhouse. Three experimental groups of animals each were tested: control, moderate temperature (MT), and high temperature (HT). MT and HT groups reached, respectively, 52.3 ± 5.1 and 63.8 ± 7.5 °C at the end of exposure. Effectiveness of the treatment was assessed by evaluating the bacteria amount of the treated abscess 1 and 4 days after treatment. Myeloperoxidase (MPO) assay evaluating the neutrophil amount was performed to assess the local neutrophil recruitment and the white blood cell count was used to evaluate the systemic inflammatory response after focused ultrasound treatment. Results: Macroscopic evaluation of treated abscess indicated a diminution of external size of abscess 1 day after treatment. Treatment did not cause open wounds. The median (lower to upper quartile) bacterial count 1 day after treatment was 6.18 × 10{sup 3} (0.76 × 10{sup 3}–11.18 × 10{sup 3}), 2.86 × 10{sup 3} (1.22 × 10{sup 3}–7.07 × 10{sup 3}), and 3.52 × 10{sup 3} (1.18 × 10{sup 3}–6.72 × 10{sup 3}) cfu/100 μl for control, MT and HT groups, respectively; for the 4-day end point, the count was 1.37 × 10{sup 3} (0.67 × 10{sup 3}–2.89 × 10{sup 3}), 1.35 × 10{sup 3} (0.09 × 10{sup 3}–2.96 × 10{sup 3}), and 0.07 × 10{sup 3} (0.03 × 10{sup 3}–0.36 × 10{sup 3}) cfu/100 μl for control, MT and HT, showing a significant reduction (p = 0.002) on the bacterial load four days after focused ultrasound treatment when treating at high temperature (HT). The MPO amount remained unchanged between groups and days, indicating no change on local neutrophil recruitment in the abscess caused by the treatment. The white blood cell count remained unchanged between groups and days indicating that no systemic inflammatory response was caused by the treatment. Conclusions: Focused ultrasound induces a therapeutic effect in abscesses induced by MRSA. This effect is observed as a reduction of the number bacteria without significantly altering the amount of MPO at the site of a MRSA-induced abscess. These initial results suggest that focused ultrasound is a viable option for the treatment of localized MRSA-related infections.

  16. Di(2-ethylhexyl) phthalate inhibits antral follicle growth, induces atresia, and inhibits steroid hormone production in cultured mouse antral follicles

    SciTech Connect (OSTI)

    Hannon, Patrick R. Brannick, Katherine E. Wang, Wei Gupta, Rupesh K. Flaws, Jodi A.

    2015-04-01

    Di(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental toxicant found in consumer products that causes ovarian toxicity. Antral follicles are the functional ovarian units and must undergo growth, survival from atresia, and proper regulation of steroidogenesis to ovulate and produce hormones. Previous studies have determined that DEHP inhibits antral follicle growth and decreases estradiol levels in vitro; however, the mechanism by which DEHP elicits these effects is unknown. The present study tested the hypothesis that DEHP directly alters regulators of the cell cycle, apoptosis, and steroidogenesis to inhibit antral follicle functionality. Antral follicles from adult CD-1 mice were cultured with vehicle control or DEHP (1–100 μg/ml) for 24–96 h to establish the temporal effects of DEHP on the follicle. Following 24–96 h of culture, antral follicles were subjected to gene expression analysis, and media were subjected to measurements of hormone levels. DEHP increased the mRNA levels of cyclin D2, cyclin dependent kinase 4, cyclin E1, cyclin A2, and cyclin B1 and decreased the levels of cyclin-dependent kinase inhibitor 1A prior to growth inhibition. Additionally, DEHP increased the mRNA levels of BCL2-associated agonist of cell death, BCL2-associated X protein, BCL2-related ovarian killer protein, B-cell leukemia/lymphoma 2, and Bcl2-like 10, leading to an increase in atresia. Further, DEHP decreased the levels of progesterone, androstenedione, and testosterone prior to the decrease in estradiol levels, with decreased mRNA levels of side-chain cleavage, 17α-hydroxylase-17,20-desmolase, 17β-hydroxysteroid dehydrogenase, and aromatase. Collectively, DEHP directly alters antral follicle functionality by inhibiting growth, inducing atresia, and inhibiting steroidogenesis. - Highlights: • DEHP inhibits antral follicle growth by dysregulating cell cycle regulators. • DEHP induces antral follicle atresia by dysregulating apoptosis regulators. • DEHP inhibits the production of antral follicle produced sex steroid hormones.

  17. DNA repair efficiency in germ cells and early mouse embryos and consequences for radiation-induced transgenerational genomic damage

    SciTech Connect (OSTI)

    Marchetti, Francesco; Wyrobek, Andrew J.

    2009-01-18

    Exposure to ionizing radiation and other environmental agents can affect the genomic integrity of germ cells and induce adverse health effects in the progeny. Efficient DNA repair during gametogenesis and the early embryonic cycles after fertilization is critical for preventing transmission of DNA damage to the progeny and relies on maternal factors stored in the egg before fertilization. The ability of the maternal repair machinery to repair DNA damage in both parental genomes in the fertilizing egg is especially crucial for the fertilizing male genome that has not experienced a DNA repair-competent cellular environment for several weeks prior to fertilization. During the DNA repair-deficient period of spermatogenesis, DNA lesions may accumulate in sperm and be carried into the egg where, if not properly repaired, could result in the formation of heritable chromosomal aberrations or mutations and associated birth defects. Studies with female mice deficient in specific DNA repair genes have shown that: (i) cell cycle checkpoints are activated in the fertilized egg by DNA damage carried by the sperm; and (ii) the maternal genotype plays a major role in determining the efficiency of repairing genomic lesions in the fertilizing sperm and directly affect the risk for abnormal reproductive outcomes. There is also growing evidence that implicates DNA damage carried by the fertilizing gamete as a mediator of postfertilization processes that contribute to genomic instability in subsequent generations. Transgenerational genomic instability most likely involves epigenetic mechanisms or error-prone DNA repair processes in the early embryo. Maternal and embryonic DNA repair processes during the early phases of mammalian embryonic development can have far reaching consequences for the genomic integrity and health of subsequent generations.

  18. Conditional inactivation of Brca1 in the mouse ovarian surface epithelium results in an increase in preneoplastic changes

    SciTech Connect (OSTI)

    Clark-Knowles, Katherine V. . E-mail: kclar075@uottawa.ca; Garson, Kenneth; Jonkers, Jos; Vanderhyden, Barbara C.

    2007-01-01

    Epithelial ovarian cancer (EOC) is thought to arise from the ovarian surface epithelium (OSE); however, the molecular events underlying this transformation are poorly understood. Germline mutations in the BRCA1 tumor suppressor gene result in a significantly increased risk of developing EOC and a large proportion of sporadic EOCs display some sort of BRCA1 dysfunction. Using mice with conditional expression of Brca1, we inactivated Brca1 in the murine OSE and demonstrate that this inactivation results in the development of preneoplastic changes, such as hyperplasia, epithelial invaginations, and inclusion cysts, which arise earlier and are more numerous than in control ovaries. These changes resemble the premalignant lesions that have been reported in human prophylactic oophorectomy specimens from women with BRCA1 germline mutation. We also report that inactivation of Brca1 in primary cultures of murine OSE cells leads to a suppression of proliferation due to increased apoptosis that can be rescued by concomitant inactivation of p53. These observations, along with our finding that these cells display an increased sensitivity to the DNA-damaging agent cisplatin, indicate that loss of function of Brca1 in OSE cells impacts both cellular growth control and DNA-damage repair which results in altered cell behavior manifested as morphological changes in vivo that arise earlier and are more numerous than what can be attributed to ageing.

  19. Engineering shallow spins in diamond with nitrogen delta-doping

    SciTech Connect (OSTI)

    Ohno, Kenichi; Joseph Heremans, F.; Bassett, Lee C.; Myers, Bryan A.; Toyli, David M.; Bleszynski Jayich, Ania C.; Palmstrom, Christopher J.; Awschalom, David D.

    2012-08-20

    We demonstrate nanometer-precision depth control of nitrogen-vacancy (NV) center creation near the surface of synthetic diamond using an in situ nitrogen delta-doping technique during plasma-enhanced chemical vapor deposition. Despite their proximity to the surface, doped NV centers with depths (d) ranging from 5 to 100 nm display long spin coherence times, T{sub 2} > 100 {mu}s at d = 5 nm and T{sub 2} > 600 {mu}s at d {>=} 50 nm. The consistently long spin coherence observed in such shallow NV centers enables applications such as atomic-scale external spin sensing and hybrid quantum architectures.

  20. Process for recovery of hydrogen and

    DOE Patents [OSTI]

    James, Brian R.; Li-Lee, Chung; Lilga, Michael A.; Nelson, David A.

    1987-01-01

    on of sulfur Abstract A process of abstracting sulfur from H.sub.2 S and generating hydrogen is disclosed comprising dissolving Pd.sub.2 X.sub.2 (.mu.-dppm).sub.2 in a solvent and then introducing H.sub.2 S. The palladium complex abstracts sulfur, forming hydrogen and a (.mu.-S) complex. The (.mu.-S) complex is readily oxidizable to a (.mu.-SO.sub.2) adduct which spontaneously loses SO.sub.2 and regenerates the palladium complex.

  1. AAV-mediated delivery of the transcription factor XBP1s into the striatum reduces mutant Huntingtin aggregation in a mouse model of Huntington's disease

    SciTech Connect (OSTI)

    Zuleta, Amparo [Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago (Chile) [Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago (Chile); Center for Molecular Studies of the Cell, Institute of Biomedical Sciences, University of Chile, Santiago (Chile); Vidal, Rene L. [Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago (Chile) [Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago (Chile); Neurounion Biomedical Foundation, Santiago (Chile); Armentano, Donna; Parsons, Geoffrey [Department of Molecular Biology, Genzyme Corporation, 49 New York Avenue, Framingham, MA 01701 (United States)] [Department of Molecular Biology, Genzyme Corporation, 49 New York Avenue, Framingham, MA 01701 (United States); Hetz, Claudio, E-mail: chetz@med.uchile.cl [Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago (Chile) [Biomedical Neuroscience Institute, Faculty of Medicine, University of Chile, Santiago (Chile); Center for Molecular Studies of the Cell, Institute of Biomedical Sciences, University of Chile, Santiago (Chile); Department of Immunology and Infectious Diseases, Harvard School of Public Health, 651 Huntington Av., Boston, MA 02446 (United States); Neurounion Biomedical Foundation, Santiago (Chile)

    2012-04-13

    Highlights: Black-Right-Pointing-Pointer The contribution of ER stress to HD has not been directly addressed. Black-Right-Pointing-Pointer Expression of XBP1s using AAVs decreases Huntingtin aggregation in vivo. Black-Right-Pointing-Pointer We describe a new in vivo model of HD based on the expression of a large fragment of mHtt-RFP. -- Abstract: Huntington's disease (HD) is caused by mutations that expand a polyglutamine region in the amino-terminal domain of Huntingtin (Htt), leading to the accumulation of intracellular inclusions and progressive neurodegeneration. Recent reports indicate the engagement of endoplasmic reticulum (ER) stress responses in human HD post mortem samples and animal models of the disease. Adaptation to ER stress is mediated by the activation of the unfolded protein response (UPR), an integrated signal transduction pathway that attenuates protein folding stress by controlling the expression of distinct transcription factors including X-Box binding protein 1 (XBP1). Here we targeted the expression of XBP1 on a novel viral-based model of HD. We delivered an active form of XBP1 locally into the striatum of adult mice using adeno-associated vectors (AAVs) and co-expressed this factor with a large fragment of mutant Htt as a fusion protein with RFP (Htt588{sup Q95}-mRFP) to directly visualize the accumulation of Htt inclusions in the brain. Using this approach, we observed a significant reduction in the accumulation of Htt588{sup Q95}-mRFP intracellular inclusion when XBP1 was co-expressed in the striatum. These results contrast with recent findings indicating a protective effect of XBP1 deficiency in neurodegeneration using knockout mice, and suggest a potential use of gene therapy strategies to manipulate the UPR in the context of HD.

  2. Bisphenol A at a low concentration boosts mouse spermatogonial cell proliferation by inducing the G protein-coupled receptor 30 expression

    SciTech Connect (OSTI)

    Sheng, Zhi-Guo; Huang, Wei; Liu, Yu-Xiang; Zhu, Ben-Zhan

    2013-02-15

    Bisphenol A (BPA) is one of the most prevalent chemicals in daily-use materials, therefore, human exposure to BPA is ubiquitous. We found that low concentrations of BPA stimulate the spermatogonial GC-1 cells proliferation by G protein-coupled receptor 30 (GPR30)-mediated epidermal growth factor receptor (EGFR)-extracellular regulated kinase (ERK)-c-Fos pathway. However, through the same pathway GPR30 expression has been shown to be induced by EGF, an EGFR ligand. Thus, we want to know if low concentrations of BPA are able to induce the GPR30 expression and the possible mechanism(s) in GC-1 cells. By transient transfection with expression plasmids, 10{sup ?9} M BPA significantly transactivates the Gpr30-5?-flanking region through activating the GPR30, cGMP-dependent protein kinase (PKG), estrogen receptor-? (ER-?), and EFGR-ERK pathways. Furthermore, an activator protein-1 (AP-1) site located within this region is found to be responsible for the transactivation of BPA. Expectedly, through the same pathways, BPA significantly induces the gene and protein expression of GPR30. c-Fos is further observed to be strongly recruited to the AP-1 site in a chromatin immunoprecipitation assay and its dysfunction on the AP-1 site markedly suppresses the expression of GPR30, p-ERK1/2, p-Ser118-ER-? and cell proliferation by BPA. Our results demonstrate that a low-concentration BPA induces GPR30 expression through the GPR30-EFGR-ERK-c-Fos, ER-?, and PKG pathways, presumably boosting the cells proliferation via a regulatory loop. The present study provides a novel insight into the potential role of GPR30 in the initiation and progression of male germ cell cancer induced by environmentally relevant BPA. - Highlights: ? Low concentrations of BPA activate the PKG and GPR30-EFGR-ERK-ER-? pathways. ? Low concentrations of BPA activate the AP-1 site of Gpr30-5?-flanking region. ? Low concentrations of BPA induce the expression of GPR30 gene and protein. ? Low concentrations of BPA boost GC-1 cells proliferation via a regulatory loop.

  3. Application of a fuzzy neural network model in predicting polycyclic aromatic hydrocarbon-mediated perturbations of the Cyp1b1 transcriptional regulatory network in mouse skin

    SciTech Connect (OSTI)

    Larkin, Andrew; Siddens, Lisbeth K.; Krueger, Sharon K.; Tilton, Susan C.; Waters, Katrina M.; Williams, David E.; Baird, William M.

    2013-03-01

    Polycyclic aromatic hydrocarbons (PAHs) are present in the environment as complex mixtures with components that have diverse carcinogenic potencies and mostly unknown interactive effects. Non-additive PAH interactions have been observed in regulation of cytochrome P450 (CYP) gene expression in the CYP1 family. To better understand and predict biological effects of complex mixtures, such as environmental PAHs, an 11 gene input-1 gene output fuzzy neural network (FNN) was developed for predicting PAH-mediated perturbations of dermal Cyp1b1 transcription in mice. Input values were generalized using fuzzy logic into low, medium, and high fuzzy subsets, and sorted using k-means clustering to create Mamdani logic functions for predicting Cyp1b1 mRNA expression. Model testing was performed with data from microarray analysis of skin samples from FVB/N mice treated with toluene (vehicle control), dibenzo[def,p]chrysene (DBC), benzo[a]pyrene (BaP), or 1 of 3 combinations of diesel particulate extract (DPE), coal tar extract (CTE) and cigarette smoke condensate (CSC) using leave-one-out cross-validation. Predictions were within 1 log{sub 2} fold change unit of microarray data, with the exception of the DBC treatment group, where the unexpected down-regulation of Cyp1b1 expression was predicted but did not reach statistical significance on the microarrays. Adding CTE to DPE was predicted to increase Cyp1b1 expression, whereas adding CSC to CTE and DPE was predicted to have no effect, in agreement with microarray results. The aryl hydrocarbon receptor repressor (Ahrr) was determined to be the most significant input variable for model predictions using back-propagation and normalization of FNN weights. - Highlights: ? Tested a model to predict PAH mixture-mediated changes in Cyp1b1 expression ? Quantitative predictions in agreement with microarrays for Cyp1b1 induction ? Unexpected difference in expression between DBC and other treatments predicted ? Model predictions for combining PAH mixtures in agreement with microarrays ? Predictions highly dependent on aryl hydrocarbon receptor repressor expression.

  4. Lupeol induces p53 and cyclin-B-mediated G2/M arrest and targets apoptosis through activation of caspase in mouse skin

    SciTech Connect (OSTI)

    Nigam, Nidhi Prasad, Sahdeo; George, Jasmine; Shukla, Yogeshwer

    2009-04-03

    Lupeol, present in fruits and medicinal plants, is a biologically active compound that has been shown to have various pharmacological properties in experimental studies. In the present study, we demonstrated the modulatory effect of lupeol on 7,12-dimethylbenz[a]anthracene (DMBA)-induced alterations on cell proliferation in the skin of Swiss albino mice. Lupeol treatment showed significant (p < 0.05) preventive effects with marked inhibition at 48, 72, and 96 h against DMBA-mediated neoplastic events. Cell-cycle analysis showed that lupeol-induced G2/M-phase arrest (16-37%) until 72 h, and these inhibitory effects were mediated through inhibition of the cyclin-B-regulated signaling pathway involving p53, p21/WAF1, cdc25C, cdc2, and cyclin-B gene expression. Further lupeol-induced apoptosis was observed, as shown by an increased sub-G1 peak (28%) at 96 h, with upregulation of bax and caspase-3 genes and downregulation of anti-apoptotic bcl-2 and survivin genes. Thus, our results indicate that lupeol has novel anti-proliferative and apoptotic potential that may be helpful in designing strategies to fight skin cancer.

  5. Exposure to Ionizing Radiation Causes Long-Term Increase in Serum Estradiol and Activation of PI3K-Akt Signaling Pathway in Mouse Mammary Gland

    SciTech Connect (OSTI)

    Suman, Shubhankar; Johnson, Michael D.; Fornace, Albert J.; Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC ; Datta, Kamal

    2012-10-01

    Purpose: Exposure to ionizing radiation is an established risk factor for breast cancer. Radiation exposure during infancy, childhood, and adolescence confers the highest risk. Although radiation is a proven mammary carcinogen, it remains unclear where it acts in the complex multistage process of breast cancer development. In this study, we investigated the long-term pathophysiologic effects of ionizing radiation at a dose (2 Gy) relevant to fractionated radiotherapy. Methods and Materials: Adolescent (6-8 weeks old; n = 10) female C57BL/6J mice were exposed to 2 Gy total body {gamma}-radiation, the mammary glands were surgically removed, and serum and urine samples were collected 2 and 12 months after exposure. Molecular pathways involving estrogen receptor-{alpha} (ER{alpha}) and phosphatidylinositol-3-OH kinase (PI3K)-Akt signaling were investigated by immunohistochemistry and Western blot. Results: Serum estrogen and urinary levels of the oncogenic estrogen metabolite (16{alpha}OHE1) were significantly increased in irradiated animals. Immunostaining for the cellular proliferative marker Ki-67 and cyclin-D1 showed increased nuclear accumulation in sections of mammary glands from irradiated vs. control mice. Marked increase in p85{alpha}, a regulatory sub-unit of the PI3K was associated with increase in Akt, phospho-Akt, phospho-BAD, phospho-mTOR, and c-Myc in irradiated samples. Persistent increase in nuclear ER{alpha} in mammary tissues 2 and 12 months after radiation exposure was also observed. Conclusions: Taken together, our data not only support epidemiologic observations associating radiation and breast cancer but also, specify molecular events that could be involved in radiation-induced breast cancer.

  6. Neurobehavioral impairments, generation of oxidative stress and release of pro-apoptotic factors after chronic exposure to sulphur mustard in mouse brain

    SciTech Connect (OSTI)

    Sharma, Deep Raj; Sunkaria, Aditya; Bal, Amanjit; Bhutia, Yangchen D.; Vijayaraghavan, R.; Flora, S.J.S.; Gill, Kiran Dip

    2009-10-15

    Recent global events have focused attention on the potential threat of international and domestic chemical terrorism, as well as the possibility of chemical warfare proliferation. Sulphur mustard (SM) is one of the potent chemical warfare agents (CWA), which initiates a cascade of events that converge on the redox mechanisms common to brain injury. The present study was designed to examine the effects of chronic SM exposure on neurobehavioral impairments, mitochondrial oxidative stress in male Swiss Albino mice and its role in inducing apoptotic neuronal cell death. The animals were divided into four groups (control, low, medium and high dose) of 5 animals each. Exposure to SM was given percutaneously daily for 12 weeks. The results demonstrated impairment in neurobehavioral indices viz. rota rod, passive avoidance and water maze tests in a dose dependent manner. There was a significant increase in lipid peroxidation and protein carbonyl content whereas, decrease in the activity of manganese superoxide dismutase (MnSOD), glutathione reductase and glutathione peroxidase suggesting impaired antioxidant defense system. Immunoblotting of cytochrome c, Bcl-2, Bax and activation of caspase-3 suggest induction of apoptosis in a dose dependent manner. Finally, increased p53 expression suggests that it may target the mitochondrial pathway for inducing apoptosis in response to DNA damage signals. In conclusion, chronic SM exposure may have the potential to generate oxidative stress which may trigger the release of cytochrome c as well as caspase-3 activation in neurons leading to cell death by apoptosis in a dose dependent manner which may in the end be responsible for the disruption of cognitive functions in mice.

  7. Heavy-Duty Low-Temperature and Diesel Combustion & Heavy-Duty Combustion

    Broader source: Energy.gov (indexed) [DOE]

    Modeling | Department of Energy 1 DOE Hydrogen and Fuel Cells Program, and Vehicle Technologies Program Annual Merit Review and Peer Evaluation PDF icon ace001_musculus_2011_o.pdf More Documents & Publications Heavy-Duty Low-Temperature and Diesel Combustion & Heavy-Duty Combustion Modeling Heavy-Duty Low-Temperature and Diesel Combustion & Heavy-Duty Combustion Modeling High Efficiency Fuel Reactivity Controlled Compression Ignition Combustion

  8. In-Cylinder Processes of EGR-Diluted Low-Load, Low-Temperature Diesel

    Broader source: Energy.gov (indexed) [DOE]

    Combustion | Department of Energy In-Cylinder Processes of EGR-Diluted Low-Load, Low-Temperature Diesel Combustion PDF icon deer09_musculus.pdf More Documents & Publications A Conceptual Model for Partially PremixedLow-Temperature Diesel Combustion Based onIn-Cylinder Laser Diagnostics and Chemical Kinetics Modeling Heavy-Duty Low-Temperature and Diesel Combustion & Heavy-Duty Combustion Modeling Visualization of UHC Emissions for Low-Temperature Diesel Engine

  9. A VME timing system for the tokamak ISTTOK

    SciTech Connect (OSTI)

    Varandas, C.A.F.; Carvalho, B.; Fernandes, H.; Sousa, J.; Cabral, J.A.C.

    1995-05-01

    This paper describes the ISTTOK timing system, which was built under a centralized philosophy based on a locally developed 16 channel, 1 {mu}s maximum resolution, VME unit. The trigger options for each channel are provided, following an innovatory approach, by a programmable pulse multiplexer. {copyright} {ital 1995} {ital American} {ital Institute} {ital of} {ital Physics}.

  10. Numerical investigation of pulse-modulated atmospheric radio frequency discharges in helium under different duty cycles

    SciTech Connect (OSTI)

    Sun Jizhong; Ding Zhengfen; Li Xuechun; Wang Dezhen [School of Physics and Optoelectronic Technology, Dalian University of Technology, Dalian 116023 (China); Wang Qi [Dalian Institute of Semiconductor Technology, School of Electronics Science and Technology, Dalian University of Technology, Dalian 116023 (China)

    2011-12-15

    Experiments observed that the pulse duty cycle has effects on the plasma homogeneity in pulse-modulated radio frequency (rf) discharges. In this paper, pulse-modulated rf (13.56 MHz) helium discharges are theoretically investigated using a two dimensional fluid model. With the pulse period being fixed to 15 {mu}s, it is found that when the pulse-on duration is over 4 {mu}s, i.e., the duty cycle is larger than approximately 27%, the discharge transits from an inhomogeneous to a homogeneous mode in every specific part of each pulse cycle under currently-used simulation parameters. More quantitative analysis shows that the discharge becomes more homogeneous as the duty cycle is increased but does not reach complete homogeneity. Possible reasons for the homogeneity improvement are discussed.

  11. Prompt-period measurement of the Annular Core Research Reactor prompt neutron generation time

    SciTech Connect (OSTI)

    Coats, R.L.; Talley, D.G.; Trowbridge, F.R.

    1994-07-01

    The prompt neutron generation time for the Annular Core Research Reactor was experimentally determined using a prompt-period technique. The resultant value of 25.5 {mu}s agreed well with the analytically determined value of 24 {mu}s. The three different methods of reactivity insertion determination yielded {+-}5% agreement in the experimental values of the prompt neutron generation time. Discrepancies observed in reactivity insertion values determined by the three methods used (transient rod position, relative delayed critical control rod positions, and relative transient rod and control rod positions) were investigated to a limited extent. Rod-shadowing and low power fuel/coolant heat-up were addressed as possible causes of the discrepancies.

  12. Development of high-voltage pulse-slicer unit with variable pulse duration for pulse radiolysis system

    SciTech Connect (OSTI)

    Upadhyay, J.; Sharma, M. L.; Navathe, C. P.; Toley, M. A.; Shinde, S. J.; Nadkarni, S. A.; Sarkar, S. K.

    2012-02-15

    A high-voltage pulse-slicer unit with variable pulse duration has been developed and integrated with a 7 MeV linear electron accelerator (LINAC) for pulse radiolysis investigation. The pulse-slicer unit provides switching voltage from 1 kV to 10 kV with rise time better than 5 ns. Two MOSFET based 10 kV switches were configured in differential mode to get variable duration pulses. The high-voltage pulse has been applied to the deflecting plates of the LINAC for slicing of electron beam of 2 {mu}s duration. The duration of the electron beam has been varied from 30 ns to 2 {mu}s with the optimized pulse amplitude of 7 kV to get corresponding radiation doses from 6 Gy to 167 Gy.

  13. Improved Measurement of the Positive-Muon Lifetime and Determination of the Fermi Constant

    SciTech Connect (OSTI)

    Chitwood, D. B.; Clayton, S. M.; Crnkovic, J.; Debevec, P. T.; Hertzog, D. W.; Kammel, P.; Kiburg, B.; Kunkle, J.; McNabb, R.; Mulhauser, F.; Oezben, C. S.; Polly, C. C.; Webber, D. M.; Winter, P.; Banks, T. I.; Crowe, K. M.; Lauss, B.; Barnes, M. J.; Wait, G. D.; Battu, S.

    2007-07-20

    The mean life of the positive muon has been measured to a precision of 11 ppm using a low-energy, pulsed muon beam stopped in a ferromagnetic target, which was surrounded by a scintillator detector array. The result, {tau}{sub {mu}}=2.197 013(24) {mu}s, is in excellent agreement with the previous world average. The new world average {tau}{sub {mu}}=2.197 019(21) {mu}s determines the Fermi constant G{sub F}=1.166 371(6)x10{sup -5} GeV{sup -2} (5 ppm). Additionally, the precision measurement of the positive-muon lifetime is needed to determine the nucleon pseudoscalar coupling g{sub P}.

  14. Method and apparatus for improved high power impulse magnetron sputtering

    DOE Patents [OSTI]

    Anders, Andre

    2013-11-05

    A high power impulse magnetron sputtering apparatus and method using a vacuum chamber with a magnetron target and a substrate positioned in the vacuum chamber. A field coil being positioned between the magnetron target and substrate, and a pulsed power supply and/or a coil bias power supply connected to the field coil. The pulsed power supply connected to the field coil, and the pulsed power supply outputting power pulse widths of greater that 100 .mu.s.

  15. Solid state switch

    DOE Patents [OSTI]

    Merritt, Bernard T.; Dreifuerst, Gary R.

    1994-01-01

    A solid state switch, with reverse conducting thyristors, is designed to operate at 20 kV hold-off voltage, 1500 A peak, 1.0 .mu.s pulsewidth, and 4500 pps, to replace thyratrons. The solid state switch is more reliable, more economical, and more easily repaired. The switch includes a stack of circuit card assemblies, a magnetic assist and a trigger chassis. Each circuit card assembly contains a reverse conducting thyristor, a resistor capacitor network, and triggering circuitry.

  16. Laser synchronized high-speed shutter for spectroscopic application

    DOE Patents [OSTI]

    Miles, Paul C.; Porter, Eldon L.; Prast, Thomas L.; Sunnarborg, Duane A.

    2002-01-01

    A fast mechanical shutter, based on rotating chopper wheels, has been designed and implemented to shutter the entrance slit of a spectrograph. This device enables an exposure time of 9 .mu.s to be achieved for a 0.8 mm wide spectrograph entrance slit, achieves 100% transmission in the open state, and an essentially infinite extinction ratio. The device further incorporates chopper wheel position sensing electronics to permit the synchronous triggering of a laser source.

  17. The development of a one microsecond pulse length, repetitively pulsed, high power modulator and a long-pulse electron beam diode for the production of intense microwaves

    SciTech Connect (OSTI)

    Stringfield, R.M.; Faehl, R.J.; Fazio, M.V.; Hoeberling, R.F.; Kwan, T.J.T.; Rickel, D.G.; VanHaaften, F.; Wasierski, R.F.; Erickson, A.; Rust, K.

    1992-07-01

    This paper discusses the pulse power and explosive emission electron beam diode development effort we have undertaken to power a relativistic klystron amplifier (RKA) microwave source. The pulsed power and electron beam must enable the RKA to Produce one kilojoule of 13 GHz radiation per pulse at a 5 Hz repetition frequency. These efforts include tests and improvements of a 1 {mu}s pulse length thyratron switched modulator, and the computational and experimental design of a 1-{mu}s-pulse-length explosive emission electron gun. The one microsecond pulse length is almost an order of magnitude beyond what has been achieved heretofore with an RKA. Achieving a peak power approaching 1 GW for 1 {mu}s requires a well behaved electron beam on that time scale. An electron beam diode has been developed that delivers a peak current of 4 to 5 kA for a pulse duration exceeding 1 {mu}s, at a beam kinetic energy above 600 keV. BANSHEE is the high voltage modulator designed for use as an electron beam driver for high power microwave tube development. The BANSHEE output pulse design parameters are 1 MV and 10 kA, with a 1 {mu}s pulse width at a repetition rate of 3--5 Hz, driving a load of impedance of 100 ohms. BANSHEE is a thyratron-switched line-type modular with a pulse transformer output stage. The modulator design is pushing the state of the art in thyratron technology and capacitor lifetime. The results of the BANSHEE modulator testing are described.

  18. Feasibility study of volumetric modulated arc therapy with constant dose rate for endometrial cancer

    SciTech Connect (OSTI)

    Yang, Ruijie; Wang, Junjie; Xu, Feng; Li, Hua; Zhang, Xile

    2013-10-01

    To investigate the feasibility, efficiency, and delivery accuracy of volumetric modulated arc therapy with constant dose rate (VMAT-CDR) for whole-pelvic radiotherapy (WPRT) of endometrial cancer. The nine-field intensity-modulated radiotherapy (IMRT), VMAT with variable dose-rate (VMAT-VDR), and VMAT-CDR plans were created for 9 patients with endometrial cancer undergoing WPRT. The dose distribution of planning target volume (PTV), organs at risk (OARs), and normal tissue (NT) were compared. The monitor units (MUs) and treatment delivery time were also evaluated. For each VMAT-CDR plan, a dry run was performed to assess the dosimetric accuracy with MatriXX from IBA. Compared with IMRT, the VMAT-CDR plans delivered a slightly greater V{sub 20} of the bowel, bladder, pelvis bone, and NT, but significantly decreased the dose to the high-dose region of the rectum and pelvis bone. The MUs decreased from 1105 with IMRT to 628 with VMAT-CDR. The delivery time also decreased from 9.5 to 3.2 minutes. The average gamma pass rate was 95.6% at the 3%/3 mm criteria with MatriXX pretreatment verification for 9 patients. VMAT-CDR can achieve comparable plan quality with significant shorter delivery time and smaller number of MUs compared with IMRT for patients with endometrial cancer undergoing WPRT. It can be accurately delivered and be an alternative to IMRT on the linear accelerator without VDR capability.

  19. Experimental Investigation of Axial and Beam-Riding Propulsive Physics with TEA CO{sub 2} laser

    SciTech Connect (OSTI)

    Kenoyer, D. A.; Salvador, I.; Myrabo, L. N.; Notaro, S. N.; Bragulla, P. W.

    2010-10-08

    A twin Lumonics K922M pulsed TEA CO{sub 2} laser system (pulse duration of approximately 100 ns FWHM spike, with optional 1 {mu}s tail, depending upon laser gas mix) was employed to experimentally measure both axial thrust and beam-riding behavior of Type no. 200 lightcraft engines, using a ballistic pendulum and Angular Impulse Measurement Device (AIMD, respectively. Beam-riding forces and moments were examined along with engine thrust-vectoring behavior, as a function of: a) laser beam lateral offset from the vehicle axis of symmetry; b) laser pulse energy ({approx}12 to 40 joules); c) pulse duration (100 ns, and 1 {mu}s); and d) engine size (97.7 mm to 161.2 mm). Maximum lateral momentum coupling coefficients (C{sub M}) of 75 N-s/MJ were achieved with the K922M laser whereas previous PLVTS laser (420 J, 18 {mu}s duration) results reached only 15 N-s/MJ--an improvement of 5x. Maximum axial C{sub M} performance with the K922M reached 225 N-s/MJ, or about {approx}3x larger than the lateral C{sub M} values. These axial C{sub M} results are sharply higher than the 120 N/MW previously reported for long pulse (e.g., 10-18 {mu}s)CO{sub 2} electric discharge lasers.

  20. A 120kV IGBT modulator for driving a pierce electron gun

    SciTech Connect (OSTI)

    Earley, L. M.; Brown, R. W.; Carlson, R. L.; Ferguson, P.; Haynes, W. B.; Kirbie, H. C.; Russell, S. J.; Sigler, F. E.; Smirnova, E. I.; Wheat, R. M.

    2004-01-01

    An IGBT modulator has been developed to drive a 120 kV, 23 A Pierce electron gun. The modulator is capable of producing pulses up to 10 {mu}s in width at repetition rates up to 10Hz with no active reset. The pulse rise time on the electron gun will be approximately 2 {mu}s and the remaining 8 {mu}s of flattop is tuned to have a ripple of less than 1 percent rms. The modulator technology was developed from a previous 50 kV prototype. The modulator consists of six boards, each with one EUPEC IGBT that drives a single common step-up transformer wound on METGLAS 2605SC cores. The six transformer cores share a common bi-filar output secondary winding. The modulator uses a fiber optic trigger system and has a high voltage cable output with an epoxy receptacle on the oil end and a ceramic receptacle on the vacuum end. The 120 kV electron gun was manufactured by MDS Co. and will be used to generate sheet electron beams from the standard pencil beam produced by the Pierce electron gun.

  1. Mapping of the receptor protein-tyrosine kinase 10 to human chromosome 1q21-q23 and mouse chromosome 1H1-5 by fluorescence in situ hybridization

    SciTech Connect (OSTI)

    Edelhoff, S.; Disteche, C.M.; Lai, C.

    1995-01-01

    Receptor protein-tyrosine kinases (PTKs) play a critical role in the transduction of signals important to cell growth, differentiation, and survival. Mutations affecting the expression of receptor PTK genes have been associated with a number of vertebrate and invertebrate developmental abnormalities, and the aberrant regulation of tyrosine phosphorylation is implicated in a variety of neoplasias. One estimate suggests that approximately 100 receptor PTK genes exist in the mammalian genome, about half of which have been identified. The tyro-10 receptor protein-tyrosine kinase, first identified in a PCR-based survey for novel tyrosine kinases in the rat nervous system, defines a new subfamily of PTKs. It exhibits a catalytic domain most closely related to those found in the trk PTK receptor subfamily, which transduces signals for nerve growth factor and the related molecules brain-derived neurotrophic factor (BDNF), neurotrophin-3, and neurotrophin-4 (NT-3 and NT-4). Trk and the related PTK receptors trkB and trkC play a critical role in the neurotrophin-dependent survival of subsets of sensory and motor neurons. The predicted tyro-10 extracellular region is, however, distinct from that of the trk subfamily and is unique except for a domain shared with the blood coagulation factors V and VIII, thought to be involved in phospholipid binding. Although tyro-10 RNA is most abundant in heart and skeletal muscle in the adult rat, it is expressed in a wide variety of tissues, including the developing and mature brain. Tyro-10 appears identical to the murine TKT sequence reported by Karn et al. and exhibits a high degree of similarity with the CaK, DDR, and Nep PTKs. A ligand for tyro-10 has not yet been identified. 10 refs., 1 fig.

  2. Quantitation of repaglinide and metabolites in mouse whole-body thin tissue sections using droplet-based liquid microjunction surface sampling-high-performance liquid chromatography-electrospray ionization tandem mass spectrometry

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Chen, Weiqi; Wang, Lifei; Van Berkel, Gary J.; Gan, Jinping; Kertesz, Vilmos

    2015-11-03

    Herein, quantitation aspects of a fully automated autosampler/HPLC-MS/MS system applied for unattended droplet-based surface sampling of repaglinide dosed thin tissue sections with subsequent HPLC separation and mass spectrometric analysis of parent drug and various drug metabolites was studied. Major organs (brain, lung, liver, kidney, muscle) from whole-body thin tissue sections and corresponding organ homogenates prepared from repaglinide dosed mice were sampled by surface sampling and by bulk extraction, respectively, and analyzed by HPLC-MS/MS. A semi-quantitative agreement between data obtained by surface sampling and that by employing organ homogenate extraction was observed. Drug concentrations obtained by the two methods followed themore » same patterns for post-dose time points (0.25, 0.5, 1 and 2 h). Drug amounts determined in the specific tissues was typically higher when analyzing extracts from the organ homogenates. Furthermore, relative comparison of the levels of individual metabolites between the two analytical methods also revealed good semi-quantitative agreement.« less

  3. NOSH-aspirin (NBS-1120), a novel nitric oxide- and hydrogen sulfide-releasing hybrid is a potent inhibitor of colon cancer cell growth in vitro and in a xenograft mouse model

    SciTech Connect (OSTI)

    Chattopadhyay, Mitali; Kodela, Ravinder; Olson, Kenneth R.; Kashfi, Khosrow

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer NOSH-aspirin is the first dual acting NO and H{sub 2}S releasing hybrid. Black-Right-Pointing-Pointer Its IC{sub 50} for cell growth inhibition is in the low nano-molar range. Black-Right-Pointing-Pointer Structure-activity studies show that the sum of the parts does not equal the whole. Black-Right-Pointing-Pointer NOSH-aspirin reduced tumor growth by 85% in mice bearing a colon cancer xenograft. -- Abstract: Nonsteroidal anti-inflammatory drugs (NSAIDs) are prototypical anti-cancer agents. However, their long-term use is associated with adverse gastrointestinal effects. Recognition that endogenous gaseous mediators, nitric oxide (NO) and hydrogen sulfide (H{sub 2}S) can increase mucosal defense mechanisms has led to the development of NO- and H{sub 2}S-releasing NSAIDs with increased safety profiles. Here we report on a new hybrid, NOSH-aspirin, which is an NO- and H{sub 2}S-releasing agent. NOSH-aspirin inhibited HT-29 colon cancer growth with IC{sub 50}s of 45.5 {+-} 2.5, 19.7 {+-} 3.3, and 7.7 {+-} 2.2 nM at 24, 48, and 72 h, respectively. This is the first NSAID based agent with such high degree of potency. NOSH-aspirin inhibited cell proliferation, induced apoptosis, and caused G{sub 0}/G{sub 1} cell cycle block. Reconstitution and structure-activity studies representing a fairly close approximation to the intact molecule showed that NOSH-aspirin was 9000-fold more potent than the sum of its parts towards growth inhibition. NOSH-aspirin inhibited ovine COX-1 more than ovine COX-2. NOSH-ASA treatment of mice bearing a human colon cancer xenograft caused a reduction in volume of 85%. Taken together, these results demonstrate that NOSH-aspirin has strong anti-cancer potential and merits further evaluation.

  4. Evaluation of delivered monitor unit accuracy of gated step-and-shoot IMRT using a two-dimensional detector array

    SciTech Connect (OSTI)

    Cheong, Kwang-Ho; Kang, Sei-Kwon; Lee, MeYeon; Kim, Su SSan; Park, SoAh; Hwang, Tae-Jin; Kim, Kyoung Ju; Oh, Do Hoon; Bae, Hoonsik; Suh, Tae-Suk

    2010-03-15

    Purpose: To overcome the problem of organ motion in intensity-modulated radiation therapy (IMRT), gated IMRT is often used for the treatment of lung cancer. In this study, the authors investigated the accuracy of the delivered monitor units (MUs) from each segment during gated IMRT using a two-dimensional detector array for user-specific verification purpose. Methods: The authors planned a 6 MV photon, seven-port step-and-shoot lung IMRT delivery. The respiration signals for gated IMRT delivery were obtained from the one-dimensional moving phantom using the real-time position management (RPM) system (Varian Medical Systems, Palo Alto, CA). The beams were delivered using a Clinac iX (Varian Medical Systems, Palo Alto, CA) with the Millennium 120 MLC. The MatriXX (IBA Dosimetry GmbH, Germany) was validated through consistency and reproducibility tests as well as comparison with measurements from a Farmer-type ion chamber. The authors delivered beams with varying dose rates and duty cycles and analyzed the MatriXX data to evaluate MU delivery accuracy. Results: There was quite good agreement between the planned segment MUs and the MUs computed from the MatriXX within {+-}2% error. The beam-on times computed from the MatriXX data were almost identical for all cases, and they matched well with the RPM beam-on and beam-off signals. A slight difference was observed between them, but it was less than 40 ms. The gated IMRT delivery demonstrated an MU delivery accuracy that was equivalent to ungated IMRT, and the delivered MUs with a gating signal agreed with the planned MUs within {+-}0.5 MU regardless of dose rate and duty cycle. Conclusions: The authors can conclude that gated IMRT is able to deliver an accurate dose to a patient during a procedure. The authors believe that the methodology and results can be transferred to other vendors' devices, particularly those that do not provide MLC log data for a verification purpose.

  5. BNL-43197 BNL-43197

    Office of Scientific and Technical Information (OSTI)

    ... Laboratory. During this period a number of improvements were made in the generator to ... injected it into a mouse and imaged the mouse with their small animal rectilinear scanner. ...

  6. Post World War II missions emerge for Y-12

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    of Sciences." "Under Hollaender, Bill and Liane Russell started a large-scale mouse-genetics project in 1947. They began to build up special mouse strains for study of the...

  7. Fermilab | Director's Policy Manual | Home

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Financial Management Freedom of Information Act Requests Inclement Weather and Snow Policy Interactions with Legislators Issues Management Maintenance MOUs Between...

  8. On Target May 2013 | Jefferson Lab

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    May 2013 May 2013 The U.S. Department of Energy's Thomas Jefferson National Accelerator Facility Mouse Medicine: System Gets Clear, 3D Brain Scans of Moving Mice Mouse Medicine: System Gets Clear, 3D Brain Scans of Moving Mice Three markers attached to the head of a mouse enable the AwakeSPECT system to obtain detailed, functional images of the brain of a conscious mouse as it moves. Setting a mouse free to roam might alarm most people, but not researchers who have developed a new imaging system

  9. SU-E-T-421: Feasibility Study of Volumetric Modulated Arc Therapy with Constant Dose Rate for Endometrial Cancer

    SciTech Connect (OSTI)

    Yang, R; Wang, J

    2014-06-01

    Purpose: To investigate the feasibility, efficiency, and delivery accuracy of volumetric modulated arc therapy with constant dose rate (VMAT-CDR) for whole-pelvic radiotherapy (WPRT) of endometrial cancer. Methods: The nine-Field intensity-modulated radiotherapy (IMRT), VMAT with variable dose-rate (VMAT-VDR), and VMAT-CDR plans were created for 9 patients with endometrial cancer undergoing WPRT. The dose distribution of planning target volume (PTV), organs at risk (OARs), and normal tissue (NT) were compared. The monitor units (MUs) and treatment delivery time were also evaluated. For each VMAT-CDR plan, a dry Run was performed to assess the dosimetric accuracy with MatriXX from IBA. Results: Compared with IMRT, the VMAT-CDR plans delivered a slightly greater V20 of the bowel, bladder, pelvis bone, and NT, but significantly decreased the dose to the high-dose region of the rectum and pelvis bone. The MUs Decreased from 1105 with IMRT to 628 with VMAT-CDR. The delivery time also decreased from 9.5 to 3.2 minutes. The average gamma pass rate was 95.6% at the 3%/3 mm criteria with MatriXX pretreatment verification for 9 patients. Conclusion: VMAT-CDR can achieve comparable plan quality with significant shorter delivery time and smaller number of MUs compared with IMRT for patients with endometrial cancer undergoing WPRT. It can be accurately delivered and be an alternative to IMRT on the linear accelerator without VDR capability. This work is supported by the grant project, National Natural; Science Foundation of China (No. 81071237)

  10. Cosmic ray neutron background reduction using localized coincidence veto neutron counting

    DOE Patents [OSTI]

    Menlove, Howard O.; Bourret, Steven C.; Krick, Merlyn S.

    2002-01-01

    This invention relates to both the apparatus and method for increasing the sensitivity of measuring the amount of radioactive material in waste by reducing the interference caused by cosmic ray generated neutrons. The apparatus includes: (a) a plurality of neutron detectors, each of the detectors including means for generating a pulse in response to the detection of a neutron; and (b) means, coupled to each of the neutrons detectors, for counting only some of the pulses from each of the detectors, whether cosmic ray or fission generated. The means for counting includes a means that, after counting one of the pulses, vetos the counting of additional pulses for a prescribed period of time. The prescribed period of time is between 50 and 200 .mu.s. In the preferred embodiment the prescribed period of time is 128 .mu.s. The veto means can be an electronic circuit which includes a leading edge pulse generator which passes a pulse but blocks any subsequent pulse for a period of between 50 and 200 .mu.s. Alternately, the veto means is a software program which includes means for tagging each of the pulses from each of the detectors for both time and position, means for counting one of the pulses from a particular position, and means for rejecting those of the pulses which originate from the particular position and in a time interval on the order of the neutron die-away time in polyethylene or other shield material. The neutron detectors are grouped in pods, preferably at least 10. The apparatus also includes means for vetoing the counting of coincidence pulses from all of the detectors included in each of the pods which are adjacent to the pod which includes the detector which produced the pulse which was counted.

  11. Recent experimental results from a long-pulse J-band relativistic klystron amplifier developmental effort

    SciTech Connect (OSTI)

    Kato, K.G.; Crouch, D.D.; Sar, D.R.; Speciale, R.A.; Carlsten, B.E.; Fazio, M.V.; Haynes, W.B.; Stringfield, R.M.

    1994-12-31

    Recent experimental results, supporting simulations, and design modeling are presented from a developmental effort to a produce a long pulse ({approximately}1{mu}s) J-band (5.85-8.2 GHz) relativistic klystron amplifier (RKA) of the high current NRL genealogy. This RKA is designed to operate at approximately 6.6 GHz, with a desired RF output {approximately}700 MW. Conversion of electron beam energy to microwave energy is obtained by a mock magnetically insulated coaxial converter which, in various incarnations, can be made to be either a cavity gap extractor or an inverse cathode.

  12. Strangelet search in Pb-Pb interactions at 158 GeV/{ital c} per nucleon

    SciTech Connect (OSTI)

    Appelquist, G.; Baglin, C.; Beringer, J.; Bohm, C.; Borer, K.; Bussiere, A.; Dittus, F.; Elsener, K.; Frei, D.; Gorodetzky, P.; Guillaud, J.P.; Hugentobler, E.; Klingenberg, R.; Linden, T.; Lohmann, K.D.; Moser, U.; Pal, T.; Pretzl, K.; Schacher, J.; Sellden, B.; Stoffel, F.; Tuominiemi, J.; Zhang, Q.P.

    1996-05-01

    The NA52 experiment searches for long-lived massive strange quark matter particles, so-called {ital strangelets}, produced in Pb-Pb collisions at a beam momentum of {ital p}{sub lab}=158 AGeV/{ital c}. Upper limits for the production of strangelets at zero degree production angle covering a mass to charge ratio up to 120 GeV/{ital c}{sup 2} and lifetimes {ital t}{sub lab}{approx_gt}1.2 {mu}s are given. The data presented here were taken during the 1994 lead beam running period at CERN. {copyright} {ital 1996 The American Physical Society.}

  13. Hi-speed versatile serial crate controller for CAMAC

    SciTech Connect (OSTI)

    Horelick, D.

    1984-10-01

    A serial crate controller, primarily for use in the SLC CAMAC control system, has been designed, and has been in use for about 2 years. The design supports a party line approach, with up to 16 crates on a single twisted pair for data transfers, plus another pair for prompt L response. The bit rate is 5 megabits/s, and complete transaction times of about 10 ..mu..s are achieved for 16-bit data transfers over cables up to 1000 feet long. One of the primary objects of the design was simplicity - there are approximately 60 chips in the two-board unit.

  14. Solid state switch

    DOE Patents [OSTI]

    Merritt, B.T.; Dreifuerst, G.R.

    1994-07-19

    A solid state switch, with reverse conducting thyristors, is designed to operate at 20 kV hold-off voltage, 1,500 A peak, 1.0 [mu]s pulsewidth, and 4,500 pps, to replace thyratrons. The solid state switch is more reliable, more economical, and more easily repaired. The switch includes a stack of circuit card assemblies, a magnetic assist and a trigger chassis. Each circuit card assembly contains a reverse conducting thyristor, a resistor capacitor network, and triggering circuitry. 6 figs.

  15. Departure Roger Anthoine

    ScienceCinema (OSTI)

    None

    2011-04-25

    Remerciements et discours du D.G. H.Schopper à l'occasion du départ de Roger Anthoine (attaché de presse), qui travaillait dans la communication et quitte le Cern après 27 ans de service. Il gardait des relations avec des médias internationaux et la presse locale; remise des cadeaux: album photo avec images des musés de Genève et un radio aviation; R.A. fait un résumé de ses activités et souvenirs et remercie ses collaborateurs

  16. SlISANA MARTINEZ Governor JOHN A. SANCHEZ Lieutenant Governor

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    SlISANA MARTINEZ Governor JOHN A. SANCHEZ Lieutenant Governor March 13, 2013 Jose Franco, Manager Carlsbad Field Office Department of Energy P.O. Box 3090 NEW MEXICO ENVIRONMENT DEPARTMENT Resource Protection Division Harold Runnels Building 1190 Saint Francis Drive (87505) P.O. Box 5469, Santa Fe, NM 87502-5469 Phone (505) 827-0419 Fax (505) 827-0310 www.nrnenv.state.l1m.us CERTIFIED MAIL - RETURN RECEIPT REQUESTED M. Farok Sharif, Project Manager Nuclear Waste Partnership LLC P.O. Box 2078

  17. Measurements of prompt radiation induced conductivity of Kapton.

    SciTech Connect (OSTI)

    Preston, Eric F.; Zarick, Thomas Andrew; Sheridan, Timothy J.; Hartman, E. Frederick; Stringer, Thomas Arthur

    2010-10-01

    We performed measurements of the prompt radiation induced conductivity in thin samples of Kapton (polyimide) at the Little Mountain Medusa LINAC facility in Ogden, UT. Three mil samples were irradiated with a 0.5 {mu}s pulse of 20 MeV electrons, yielding dose rates of 1E9 to 1E10 rad/s. We applied variable potentials up to 2 kV across the samples and measured the prompt conduction current. Analysis rendered prompt conductivity coefficients between 6E-17 and 2E-16 mhos/m per rad/s, depending on the dose rate and the pulse width.

  18. Recent Upgrade of the Klystron Modulator at SLAC

    SciTech Connect (OSTI)

    Nguyen, M.N.; Burkhart, C.P.; Lam, B.K.; Morris, B.; /SLAC

    2011-11-04

    The SLAC National Accelerator Laboratory employs 244 klystron modulators on its two-mile-long linear accelerator that has been operational since the early days of the SLAC establishment in the sixties. Each of these original modulators was designed to provide 250 kV, 262 A and 3.5 {mu}S at up to 360 pps using an inductance-capacitance resonant charging system, a modified type-E pulse-forming network (PFN), and a pulse transformer. The modulator internal control comprised of large step-start resistor-contactors, vacuum-tube amplifiers, and 120 Vac relays for logical signals. A major, power-component-only upgrade, which began in 1983 to accommodate the required beam energy of the SLAC Linear Collider (SLC) project, raised the modulator peak output capacity to 360 kV, 420 A and 5.0 {mu}S at a reduced pulse repetition rate of 120 pps. In an effort to improve safety, performance, reliability and maintainability of the modulator, this recent upgrade focuses on the remaining three-phase AC power input and modulator controls. The upgrade includes the utilization of primary SCR phase control rectifiers, integrated fault protection and voltage regulation circuitries, and programmable logic controllers (PLC) -- with an emphasis on component physical layouts for safety and maintainability concerns. In this paper, we will describe the design and implementation of each upgraded component in the modulator control system. We will also report the testing and present status of the modified modulators.

  19. Multiple protocol fluorometer and method

    DOE Patents [OSTI]

    Kolber, Zbigniew S.; Falkowski, Paul G.

    2000-09-19

    A multiple protocol fluorometer measures photosynthetic parameters of phytoplankton and higher plants using actively stimulated fluorescence protocols. The measured parameters include spectrally-resolved functional and optical absorption cross sections of PSII, extent of energy transfer between reaction centers of PSII, F.sub.0 (minimal), F.sub.m (maximal) and F.sub.v (variable) components of PSII fluorescence, photochemical and non-photochemical quenching, size of the plastoquinone (PQ) pool, and the kinetics of electron transport between Q.sub.a and PQ pool and between PQ pool and PSI. The multiple protocol fluorometer, in one embodiment, is equipped with an excitation source having a controlled spectral output range between 420 nm and 555 nm and capable of generating flashlets having a duration of 0.125-32 .mu.s, an interval between 0.5 .mu.s and 2 seconds, and peak optical power of up to 2 W/cm.sup.2. The excitation source is also capable of generating, simultaneous with the flashlets, a controlled continuous, background illumination.

  20. Understanding composite explosive energetics: 3, Reactive flow modeling of aluminum reaction kinetics in PETN and TNT

    SciTech Connect (OSTI)

    Tao, W.C.; Tarver, C.M.; Ornellas, D.L.

    1991-12-06

    Using Fabry-Perot interferometry techniques, we have determined that early time rate of energy release from detonating PETN and TNT explosives filled with 5 and 10 wt % of either 5 {mu}m of 18 {mu}m spherical aluminum (Al) particles. From the measured particle velocity data, we are able to infer the reaction rate of aluminum with the detonation products, and calculate the extent of reaction 1--3 {mu}s after the detonation. We observed that a substantional portion of the aluminum metal in all of the PETN and TNE formulations reacted within the timeframe of the one-dimensional experiment. In the PETN formulation filed with 5 wt % of 5 {mu}m aluminum, all of the metal reacted within 1.5 {mu}s, resulting in an increase of 22% in energy compared to pure PETN. A reactive-flow hydrodynamic model based on the Zeldovich-von Neumann-Doring (ZND) description of the reaction zone and subsequent reaction produce expansion (Taylor wave) is used to interpret the reaction rate of the aluminum particles with detonation product gases. The diffusion-controlled reaction mechanism for aluminum and the global kinetic parameters used in the model have been found to be consistent for all the PETN and TNT formulations.

  1. Understanding composite explosive energetics: 4. Reactive flow modeling of aluminum reaction kinetics in PETN and TNT using normalized product equation of state

    SciTech Connect (OSTI)

    Tao, W.C.; Tarver, C.M.; Kury, J.W.; Lee, C.G.; Ornellas, D.L.

    1993-07-01

    Using Fabry-Perot interferometry techniques, we have determined the early time rate of energy release from detonating PETN and TNT explosives filled with 5 to 20 wt % of either 5 {mu}m or 18 {mu}m spherical aluminum with the detonation products, and calculate the extent of reaction at 1--3 {mu}s after the detonation. All of the metal in PETN formulations filled with 5 wt % and 10 wt % of either 5 {mu}m or 18 {mu}m aluminum reacted within 1.5 {mu}s, resulting in an increase of 18--22% in energy compared to pure PETN. For TNT formulations, between 5 to 10 wt % aluminum reacts completely with the same timeframe. A reactive flow hydrodynamic code model based on the Zeldovich-von Neumann-Doring (ZND) description of the reaction zone and subsequent reaction product expansion (Taylor wave) is used to address the reaction rate of the aluminum particles with detonation product gases. The detonation product JWL equation of state is derived from that of pure PETN using a parametric normalization methodology.

  2. Laser-induced temperature jump/time-resolved infrared study of the fast events in protein folding

    SciTech Connect (OSTI)

    Woodruff, W.H.; Dyer, R.B.; Williams, S. [Los Alamos National Laboratory, NM (United States); Callender, H.; Gilmanshin, R. [CUNY, NY (United States)

    1996-10-01

    Laser-induced temperature jump followed by time-resolved infrared probe of reaction dynamics are used to study the temporal evolution of polypeptide structure during protein folding and unfolding. Reactions are initiated in times of 50 ps or longer by T-jumps of 10`s of degrees, obtained by laser excitation of water overtone absorbances. Observation of the Amide I transient absorbances reveal melting lifetimes of helices unconstrained by tertiary structure to be ca. 160 ns in a model 21-peptide and ca. 30 ns in {open_quotes}molten globule{close_quotes} apomyoglobin. No other processes are observed in these systems over the timescale 50 ps to 2 ms. Equilibrium data suggest the corresponding helix formation lifetimes to be ca. 16 and 1 ns, respectively. In {open_quotes}native{close_quotes} apomyoglobin two helix melting lifetimes are observed and we infer that a third occurs on a timescale inaccessible to our experiment (> 1 ms). The shorter observed lifetime, as in the molten globule, is ca. 30 ns. The longer lifetime is ca. 70 {mu}s. We suggest that the slower process is helix melting that is rate-limited by the unfolding of tertiary structure. Equilibrium data suggest a lifetime of ca. 1 {mu}s for the development of these tertiary folds.

  3. Shock initiation sensitivity of hexanitrostilbene (HNS)

    SciTech Connect (OSTI)

    Schwarz, A C

    1981-01-01

    Experiments were conducted to study the influence of powder morphology, sample density, diameter of the impacting flyer and duration of the stimulus on the shock initiation sensitivity of the high explosive, hexanitrostilbene. The shock stimulus was provided by a polyimide flyer accelerated by an electrically-exploded, metallic foil. Impact pressure (P) was controlled between 3.8 and 9.8 GPa and pulse duration (tau) was nominally 0.035 ..mu..s except for the last experiment where pulse duration varied. Powder morphology significantly influenced the shock amplitude required for initiation with the finest-particle HNS requiring the least pressure, 6.3 GPa, and exhibiting the sharpest threshold. Both a reduction in density of HNS, from 1.60 to 1.30 Mg/m/sup 3/, and an increase in flyer diameter affected a reduction in impact velocity (or pressure) needed to induce detonation. The line which separates detontion from non-detonation is expressed by P/sup 2/ /sup 4/ tau = constant for tau between 0.01 and 0.10 ..mu..s; for longer pulses the initiation criterion becomes one of constant pressure.

  4. The LEB to MEB transfer kicker system prototype

    SciTech Connect (OSTI)

    Pappas, C.; Wilson, M.; Anderson, D.

    1994-08-01

    The design requirements for the Low Energy Booster (LEB) extraction kicker system at the Superconducting Super Collider Laboratory (SSCL) were to deflect a 12 GeV/c beam through an angle of 1.5 mrad. The circumference of the LEB was 540 M. This resulted in a 0.06 T-m integrated field, of 1.8 {mu}s width with a 1% to 99% rise time of less than 80 ns and allowable pulse ripple of less than {plus_minus}1%. The repetition frequency was 10 Hz and the allowable timing jitter was 2 ns. The field was required to be uniform over a 2{times}4 cm area to {plus_minus}2.5%. The requirements for the Medium Energy Booster (MEB) injection kicker were similar except that a 99% to 1% pulse fall time of less than 2 {mu}s was needed. Prototypes of the pulsed power system and magnet to meet these requirements were built and tested at the SSCL. This paper describes the results of that testing.

  5. Wide-range voltage modulation

    SciTech Connect (OSTI)

    Rust, K.R.; Wilson, J.M.

    1992-06-01

    The Superconducting Super Collider`s Medium Energy Booster Abort (MEBA) kicker modulator will supply a current pulse to the abort magnets which deflect the proton beam from the MEB ring into a designated beam stop. The abort kicker will be used extensively during testing of the Low Energy Booster (LEB) and the MEB rings. When the Collider is in full operation, the MEBA kicker modulator will abort the MEB beam in the event of a malfunction during the filling process. The modulator must generate a 14-{mu}s wide pulse with a rise time of less than 1 {mu}s, including the delay and jitter times. It must also be able to deliver a current pulse to the magnet proportional to the beam energy at any time during ramp-up of the accelerator. Tracking the beam energy, which increases from 12 GeV at injection to 200 GeV at extraction, requires the modulator to operate over a wide range of voltages (4 kV to 80 kV). A vacuum spark gap and a thyratron have been chosen for test and evaluation as candidate switches for the abort modulator. Modulator design, switching time delay, jitter and pre-fire data are presented.

  6. System Provides Clear Brain Scans of Awake, Unrestrained Mice...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    System Provides Clear Brain Scans of Awake, Unrestrained Mice dynamic imaging of mice ... obtain detailed, functional images of the brain of a conscious mouse as it moves around. ...

  7. Web-enabled Milestones Chart | OSTI, US Dept of Energy, Office...

    Office of Scientific and Technical Information (OSTI)

    Web-enabled Milestones Chart Over the last decade, numerous milestones have been achieved, including many "firsts" in government web search technology. Mouse over the year to view ...

  8. Sandia National Laboratories: Research: High Consequence, Automation...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    team knew they needed a robot for the job and called Sandia's High Consequence, Automation, & Robotics (HCAR) team. Mighty Mouse Challenge Typically the cylinder moved back...

  9. Transmission/Resource Library/NEPA | Open Energy Information

    Open Energy Info (EERE)

    Library Jump to: navigation, search ResourceLibraryHeader.png Planning Public Involvement GIS Tools and Maps Environmental Resources and Mitigation NEPA MOUs General...

  10. Category:RAPID Best Practices | Open Energy Information

    Open Energy Info (EERE)

    Practices RAPIDBest PracticesCoordinating Permit Offices RAPIDBest PracticesLandscape-Scale Mitigation R cont. RAPIDBest PracticesMemorandums of Understanding (MOUs)...

  11. Final Technical Report

    SciTech Connect (OSTI)

    Bult Carol J.

    2003-11-24

    The results of the DOE-funded Mouse Genome Sequence (MGS) project include a significant enhancement in the capacity of the community to connect biological knowledge with the mouse genome sequence in a comparative context. The resources developed as the result of the activities of the MGS project staff are used extensively by both individual researchers and other informatics groups.

  12. Spatiotemporal temperature and density characterization of high-power atmospheric flashover discharges over inert poly(methyl methacrylate) and energetic pentaerythritol tetranitrate dielectric surfaces

    SciTech Connect (OSTI)

    Tang, V.; Grant, C. D.; McCarrick, J. F.; Zaug, J. M.; Glascoe, E. A.; Wang, H.

    2012-03-01

    A flashover arc source that delivered up to 200 mJ on the 100s-of-ns time-scale to the arc and a user-selected dielectric surface was characterized for studying high-explosive kinetics under plasma conditions. The flashover was driven over thin pentaerythritol tetranitrate (PETN) and poly(methyl methacrylate) (PMMA) dielectric films and the resultant plasma was characterized in detail. Time- and space-resolved temperatures and electron densities of the plasma were obtained using atomic emission spectroscopy. The hydrodynamics of the plasma was captured through fast, visible imaging. Fourier transform infrared spectroscopy (FTIR) was used to characterize the films pre- and post-shot for any chemical alterations. Time-resolved infrared spectroscopy (TRIR) provided PETN depletion data during the plasma discharge. For both types of films, temperatures of 1.6-1.7 eV and electron densities of {approx}7-8 x 10{sup 17}/cm{sup 3}{approx}570 ns after the start of the discharge were observed with temperatures of 0.6-0.7 eV persisting out to 15 {mu}s. At 1.2 {mu}s, spatial characterization showed flat temperature and density profiles of 1.1-1.3 eV and 2-2.8 x 10{sup 17}/cm{sup 3} for PETN and PMMA films, respectively. Images of the plasma showed an expanding hot kernel starting from radii of {approx}0.2 mm at {approx}50 ns and reaching {approx}1.1 mm at {approx}600 ns. The thin films ablated or reacted several hundred nm of material in response to the discharge. First TRIR data showing the in situ reaction or depletion of PETN in response to the flashover arc were successfully obtained, and a 2-{mu}s, 1/e decay constant was measured. Preliminary 1 D simulations compared reasonably well with the experimentally determined plasma radii and temperatures. These results complete the first steps to resolving arc-driven PETN reaction pathways and their associated kinetic rates using in situ spectroscopy techniques.

  13. SU-E-T-11: A Dosimetric Comparison of Robotic Prostatic Radiosugery Using Multi- Leaf Collimation Vs Circular Collimators

    SciTech Connect (OSTI)

    Feng, J; Yang, J; Lamond, J; Lavere, N; Laciano, R; Ding, W; Arrigo, S; Brady, L

    2014-06-01

    Purpose: The study compared the dosimetry plans of Stereotatic Body Radiotherapy (SBRT) prostate cancer patients using the M6 Cyberknife with Multi-leaf Collimation (MLC) compared with the plans using G4 Cyberknife with circular collimators. Methods: Eight previously treated prostate cancer patients' SBRT plans using circular collimators, designed with Multiplan v3.5.3, were used as a benchmark. The CT, contours and the optimization scripts were imported into Multiplan v5.0 system and replanned with MLC. The same planning objectives were used: more than 95% of PTV received 36.25Gy, 90% of prostate received 40Gy and maximum dose <45Gy, in five fractions. For organs at risk, less than 1cc of rectum received 36Gy and less than 10cc of bladder received 37Gy. Plans were evaluated on parameters derived from dose volume. The beam number, MU and delivery time were recorded to compare the treatment efficiency. Results: The mean CTV volume was 41.3cc (27.5?57.6cc) and mean PTV volume was 76.77cc (59.1?99.7cc). The mean PTV coverage was comparable between MLC (98.87%) and cone (98.74%). MLC plans had a slightly more favorable homogeneity index (1.22) and conformity index (1.17), than the cone (1.24 and 1.15). The mean rectum volume of 36 Gy (0.52cc) of MLC plans was slightly larger than cone (0.38cc) and the mean bladder volume of 37 Gy was smaller in MLC (1.82cc) than in cone plans (3.09cc). The mean number of nodes and beams were 65.9 and 80.5 in MLC vs 65.9 and 203.6 in cone. The mean MUs were significantly less for MLC plans (24,228MUs) than cone (32,347MUs). The total delivery time (which included 5 minutes for setup) was less, 29.6min (26?32min) for MLC vs 45min (35?55min) for cone. Conclusion: While the differences in the dosimetry between the MLC and circular collimator plans were rather minor, the MLC plans were much more efficient and required significantly less treatment time.

  14. SU-E-T-606: A Novel Integrated VMAT/IMRT Technique For the Treatment of Non-Small Cell Lung Cancer

    SciTech Connect (OSTI)

    Zhao, N; Yang, R; Wang, J

    2014-06-01

    Purpose: To investigate a novel Integrated VMAT/IMRT technique which combines volumetric modulated arc therapy (VMAT) and intensity modulated radiation therapy (IMRT) for non-small cell lung cancer (NSCLC). Methods: 2 partial arcs VMAT, 5-field IMRT and Integrated VMAT/IMRT plans were created for 17 patients with NSCLC. The Integrated VMAT/IMRT technique consisted of 2 partial VMAT arcs and 5 IMRT fields. The dose distribution of planning target volume (PTV) and organs at risk (OARs) for Integrated VMAT/IMRT was compared with IMRT and VMAT. The monitor units (MUs) and treatment delivery time were also evaluated. For each plan, a dry run was performed to assess the dosimetric accuracy with MatriXX from IBA. Results: Integrated VMAT/IMRT significantly improved the target conformity and homogeneity. The V30 of normal lung for Integrated plans was significantly lower than IMRT plans (8.4% vs 9.2%; p<0.05). The V5 and mean lung dose (MLD) of normal lung for Integrated plans were 9.8% and 4.6% lower than VMAT plans (p<0.05). The maximum dose of spinal cord for Integrated plans was 4.9 Gy lower than IMRT plans (p<0.05). The mean delivery time of IMRT, VMAT and Integrated plans was 280 s, 114 s, and 327 s, respectively. The mean MUs needed for IMRT, VMAT and Integrated plans were 933, 512, and 737, respectively. The gamma pass rates were beyond 90% at the 3%/3 mm criteria when the gantry angles were set to 0 for pretreatment verification. Conclusion: Integrated VMAT/IMRT technique significantly reduced V5, V10 and MLD of normal lung compared with VMAT, and the irradiated volume of the OARs receiving medium to high dose with fewer MUs compared with IMRT. Integrated VMAT/IMRT technique can be a feasible radiotherapy technique with better plan quality and accurately delivered on the linear accelerator. Ruijie Yang was funded by the grant project: National Natural Science Foundation of China (No. 81071237). Other authors have no competing interest for this work.

  15. ABSOLUTE TIMING OF THE CRAB PULSAR WITH THE INTEGRAL/SPI TELESCOPE

    SciTech Connect (OSTI)

    Molkov, S.; Jourdain, E.; Roques, J. P.

    2010-01-01

    We have investigated the pulse shape evolution of the Crab pulsar emission in the hard X-ray domain of the electromagnetic spectrum. In particular, we have studied the alignment of the Crab pulsar phase profiles measured in the hard X-rays and in other wavebands. To obtain the hard X-ray pulse profiles, we have used six years (2003-2009, with a total exposure of about 4 Ms) of publicly available data of the SPI telescope on-board the International Gamma-Ray Astrophysics Laboratory observatory, folded with the pulsar time solution derived from the Jodrell Bank Crab Pulsar Monthly Ephemeris. We found that the main pulse in the hard X-ray 20-100 keV energy band leads the radio one by 8.18 +- 0.46 milliperiods in phase, or 275 +- 15 mus in time. Quoted errors represent only statistical uncertainties. Our systematic error is estimated to be approx40 mus and is mainly caused by the radio measurement uncertainties. In hard X-rays, the average distance between the main pulse and interpulse on the phase plane is 0.3989 +- 0.0009. To compare our findings in hard X-rays with the soft 2-20 keV X-ray band, we have used data of quasi-simultaneous Crab observations with the proportional counter array monitor on-board the Rossi X-Ray Timing Explorer mission. The time lag and the pulses separation values measured in the 3-20 keV band are 0.00933 +- 0.00016 (corresponding to 310 +- 6 mus) and 0.40016 +- 0.00028 parts of the cycle, respectively. While the pulse separation values measured in soft X-rays and hard X-rays agree, the time lags are statistically different. Additional analysis show that the delay between the radio and X-ray signals varies with energy in the 2-300 keV energy range. We explain such a behavior as due to the superposition of two independent components responsible for the Crab pulsed emission in this energy band.

  16. Dosimetric comparison of different multileaf collimator leaves in treatment planning of intensity-modulated radiotherapy for cervical cancer

    SciTech Connect (OSTI)

    Wang, Shichao; Ai, Ping; Xie, Li; Xu, Qingfeng; Bai, Sen; Lu, You; Li, Ping; Chen, Nianyong

    2013-01-01

    To study the effect of multileaf collimator (MLC) leaf widths (standard MLC [sMLC] width of 10 mm and micro-MLC [mMLC] width of 4 mm) on intensity-modulated radiotherapy (IMRT) for cervical cancer. Between January 2010 and August 2010, a retrospective analysis was conducted on 12 patients with cervical cancer. The treatment plans for all patients were generated with the same machine setup parameters and optimization methods in a treatment planning system (TPS) based on 2 commercial Elekta MLC devices. The dose distribution for the planning tumor volume (PTV), the dose sparing for organs at risk (OARs), the monitor units (MUs), and the number of IMRT segments were evaluated. For the delivery efficiency, the MUs were significantly higher in the sMLC-IMRT plan than in the mMLC-IMRT plan (802 ± 56.9 vs 702 ± 56.7; p < 0.05). The number of segments in the plans were 58.75 ± 1.8 and 59 ± 1.04 (p > 0.05). For the planning quality, the conformity index (CI) between the 2 paired IMRT plans with the mMLC and the sMLC did not differ significantly (average: 0.817 ± 0.024 vs 0.810 ± 0.028; p > 0.05). The differences of the homogeneity index (HI) between the 2 paired plans were statistically significant (average: 1.122 ± 0.010 vs 1.132 ± 0.014; p < 0.01). For OARs, the rectum, bladder, small intestine, and bony pelvis were evaluated in terms of V{sub 10}, V{sub 20}, V{sub 30}, and V{sub 40}, percentage of contoured OAR volumes receiving 10, 20, 30, and 40 Gy, respectively, and the mean dose (D{sub mean}) received. The IMRT plans with the mMLC protected the OARs better than the plans with the sMLC. There were significant differences (p < 0.05) in evaluated parameters between the 2 paired IMRT plans, except for V{sub 30} and V{sub 40} of the rectum and V{sub 10}, V{sub 20}, V{sub 40}, and D{sub mean} of the bladder. IMRT plans with the mMLC showed advantages over the plans with the sMLC in dose homogeneity for targets, dose sparing of OARs, and fewer MUs in cervical cancer.

  17. Volumetric-modulated arc radiotherapy for pancreatic malignancies: Dosimetric comparison with sliding-window intensity-modulated radiotherapy and 3-dimensional conformal radiotherapy

    SciTech Connect (OSTI)

    Nabavizadeh, Nima Simeonova, Anna O.; Waller, Joseph G.; Romer, Jeanna L.; Monaco, Debra L.; Elliott, David A.; Tanyi, James A.; Fuss, Martin; Thomas, Charles R.; Holland, John M.

    2014-10-01

    Volumetric-modulated arc radiotherapy (VMAT) is an iteration of intensity-modulated radiotherapy (IMRT), both of which deliver highly conformal dose distributions. Studies have shown the superiority of VMAT and IMRT in comparison with 3-dimensional conformal radiotherapy (3D-CRT) in planning target volume (PTV) coverage and organs-at-risk (OARs) sparing. This is the first study examining the benefits of VMAT in pancreatic cancer for doses more than 55.8 Gy. A planning study comparing 3D-CRT, IMRT, and VMAT was performed in 20 patients with pancreatic cancer. Treatments were planned for a 25-fraction delivery of 45 Gy to a large field followed by a reduced-volume 8-fraction external beam boost to 59.4 Gy in total. OARs and PTV doses, conformality index (CI) deviations from 1.0, monitor units (MUs) delivered, and isodose volumes were compared. IMRT and VMAT CI deviations from 1.0 for the large-field and the boost plans were equivalent (large field: 0.032 and 0.046, respectively; boost: 0.042 and 0.037, respectively; p > 0.05 for all comparisons). Both IMRT and VMAT CI deviations from 1.0 were statistically superior to 3D-CRT (large field: 0.217, boost: 0.177; p < 0.05 for all comparisons). VMAT showed reduction of the mean dose to the boost PTV (VMAT: 61.4 Gy, IMRT: 62.4 Gy, and 3D-CRT: 62.3 Gy; p < 0.05). The mean number of MUs per fraction was significantly lower for VMAT for both the large-field and the boost plans. VMAT delivery time was less than 3 minutes compared with 8 minutes for IMRT. Although no statistically significant dose reduction to the OARs was identified when comparing VMAT with IMRT, VMAT showed a reduction in the volumes of the 100% isodose line for the large-field plans. Dose escalation to 59.4 Gy in pancreatic cancer is dosimetrically feasible with shorter treatment times, fewer MUs delivered, and comparable CIs for VMAT when compared with IMRT.

  18. Observation of Stueckelberg oscillations in dipole-dipole interactions

    SciTech Connect (OSTI)

    Ditzhuijzen, C. S. E. van; Tauschinsky, Atreju; Van Linden van den Heuvell, H. B.

    2009-12-15

    We have observed Stueckelberg oscillations in the dipole-dipole interaction between Rydberg atoms with an externally applied radio-frequency field. The oscillating rf field brings the interaction between cold Rydberg atoms in two separated volumes into resonance. We observe multiphoton transitions when varying the amplitude of the rf field and the static electric field offset. The angular momentum states we use show a quadratic Stark shift, which leads to a fundamentally different behavior than linearly shifting states. Both cases are studied theoretically using the Floquet approach and are compared. The amplitude of the sidebands, related to the interaction strength, is given by the Bessel function in the linearly shifting case and by the generalized Bessel function in the quadratically shifting case. The oscillatory behavior of both functions corresponds to Stueckelberg oscillations, an interference effect described by the semiclassical Landau-Zener-Stueckelberg model. The measurements prove coherent dipole-dipole interaction during at least 0.6 mus.

  19. Ionization of Rb Rydberg atoms in the attractive nsnp dipole-dipole potential

    SciTech Connect (OSTI)

    Park, Hyunwook; Shuman, E. S.; Gallagher, T. F.

    2011-11-15

    We have observed the ionization of a cold gas of Rb Rydberg atoms which occurs when nsns van der Waals pairs of ns atoms of n{approx_equal} 40 on a weakly repulsive potential are transferred to an attractive dipole-dipole nsnp potential by a microwave transition. Comparing the measurements to a simple model shows that the initial 300-{mu}K thermal velocity of the atoms plays an important role. Excitation to a repulsive dipole-dipole potential does not lead to more ionization on a 15-{mu}s time scale than leaving the atoms in the weakly repulsive nsns state. This observation is slightly surprising since a radiative transition must occur to allow ionization in the latter case. Finally, by power broadening of the microwave transition, to allow transitions from the initial nsns state to the nsnp state over a broad range of internuclear spacings, it is possible to accelerate markedly the evolution to a plasma.

  20. Direct Observation of the Phenomenology of a Solid Thermal Explosion Using Time-Resolved Proton Radiography

    SciTech Connect (OSTI)

    Smilowitz, L.; Henson, B. F.; Romero, J. J.; Asay, B. W.; Schwartz, C. L.; Saunders, A.; Merrill, F. E.; Morris, C. L.; Kwiatkowski, K.; Hogan, G.; Nedrow, P.; Murray, M. M.; Thompson, T. N.; McNeil, W.; Rightley, P.; Marr-Lyon, M.

    2008-06-06

    We present a new phenomenology for burn propagation inside a thermal explosion based on dynamic radiography. Radiographic images were obtained of an aluminum cased solid cylindrical sample of a plastic bonded formulation of octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine. The phenomenology observed is ignition followed by cracking in the solid accompanied by the propagation of a radially symmetric front of increasing proton transmission. This is followed by a further increase in transmission through the sample, ending after approximately 100 {mu}s. We show that these processes are consistent with the propagation of a convective burn front followed by consumption of the remaining solid by conductive particle burning.

  1. Deployment at the Savannah River Site of a standardized, modular transportable and connectable hazard category 2 nuclear system for repackaging TRU waste

    SciTech Connect (OSTI)

    Lussiez, G.; Hickman, S.; Anast, K. R.; Oliver, W. B.

    2004-01-01

    This paper describes the conception, design, fabrication and deployment of a modular, transportable, connectable Category 2 nuclear system deployed at the Savannah River site to be used for characterizing and repackaging Transuranic Waste destined for the Waste Isolation Pilot Plant (WIPP). A standardized Nuclear Category 2 and Performance Category 2 envelope called a 'Nuclear Transportainer' was conceived and designed that provides a safety envelope for nuclear operations. The Nuclear Transportainer can be outfitted with equipment that performs functions necessary to meet mission objectives, in this case repackaging waste for shipment to WIPP. Once outfitted with process and ventilation systems the Nuclear Transportainer is a Modular Unit (MU). Each MU is connectable to other MUS - nuclear or non-nuclear - allowing for multiple functions, command & control, or increasing capacity. The design took advantage of work already in-progress at Los Alamos National Laboratory (LANL) for a similar system to be deployed at LANL's Technical Area 54.

  2. A digital optical phase-locked loop for diode lasers based on field programmable gate array

    SciTech Connect (OSTI)

    Xu Zhouxiang; Zhang Xian; Huang Kaikai; Lu Xuanhui

    2012-09-15

    We have designed and implemented a highly digital optical phase-locked loop (OPLL) for diode lasers in atom interferometry. The three parts of controlling circuit in this OPLL, including phase and frequency detector (PFD), loop filter and proportional integral derivative (PID) controller, are implemented in a single field programmable gate array chip. A structure type compatible with the model MAX9382/MCH12140 is chosen for PFD and pipeline and parallelism technology have been adapted in PID controller. Especially, high speed clock and twisted ring counter have been integrated in the most crucial part, the loop filter. This OPLL has the narrow beat note line width below 1 Hz, residual mean-square phase error of 0.14 rad{sup 2} and transition time of 100 {mu}s under 10 MHz frequency step. A main innovation of this design is the completely digitalization of the whole controlling circuit in OPLL for diode lasers.

  3. Suppression of beam induced pulse shortening modes in high power RF generator TW output structures

    SciTech Connect (OSTI)

    Haimson, J.; Mecklenburg, B.

    1992-12-31

    Several different style 11.4 GHz relativistic klystrons, operating with beam pulse widths of 50 ns and using large aperture, tapered phase-velocity TW structures,` have recently demonstrated output RF power levels in the range of 100 to 300 MW without breakdown or pulse shortening. To extend this performance into the long pulse regime (1 {mu}s) or to demonstrate a threefold increase in output power by using higher currents, the existing TW circuit designs must be modified (a) to reduce the cavity maximum surface E-fields by a factor of 2 to 3, and (b) to elevate the current threshold values of the beam induced higher order modes (HOM) to ensure avoidance of RF pulse shortening and associated instabilities. A technique for substantially elevating this threshold current is described, and microwave data and photographs are presented showing the degree of HOM damping achieved in a recently constructed 11.4 GHz TW structure.

  4. Ultra-short ion and neutron pulse production

    DOE Patents [OSTI]

    Leung, Ka-Ngo; Barletta, William A.; Kwan, Joe W.

    2006-01-10

    An ion source has an extraction system configured to produce ultra-short ion pulses, i.e. pulses with pulse width of about 1 .mu.s or less, and a neutron source based on the ion source produces correspondingly ultra-short neutron pulses. To form a neutron source, a neutron generating target is positioned to receive an accelerated extracted ion beam from the ion source. To produce the ultra-short ion or neutron pulses, the apertures in the extraction system of the ion source are suitably sized to prevent ion leakage, the electrodes are suitably spaced, and the extraction voltage is controlled. The ion beam current leaving the source is regulated by applying ultra-short voltage pulses of a suitable voltage on the extraction electrode.

  5. Aggregation behavior of hexaoxyethyleneglycol myristate and hexaoxyethyleneglycol mono (1-methyltridecane) ether and dynamics of their micelles in aqueous solution

    SciTech Connect (OSTI)

    Alami, E.; Zana, R. ); Van Os, N.M.; Jong, B. de; Kerkhof, F.J.M. ); Rupert, L.A.M. )

    1993-10-01

    The title surfactants have similar critical micelle concentrations and cloud temperatures. Their micellar solutions have been investigated by time resolved fluorescence quenching in the range 2--25 c. The micelle aggregation numbers of both surfactants do not differ much, and increase with temperature. Aggregation numbers are large, suggesting anisotropic micelles, and the results show that the micelles are polydisperse. Fast intermicellar exchange of material becomes detectable on the fluorescence timescale ([approximately]1 [mu]s) above T [approx] 10 C, i.e., some 35--40 C below the cloud temperature of the solution. This exchange probably occurs via micelle collisions with temporary merging. Overall the behavior of these two surfactants is very similar to that of the other ethoxylated nonionic surfactants previously examined.

  6. Recent progress in PNC`s MLIS Project

    SciTech Connect (OSTI)

    Yamaguchi, Hiromi; Suto, Osamu; Tashiro, Kiyoshi; Kawakami, Shigeaki; Shimazaki, Yoshihiro

    1994-12-31

    Research on the molecular laser isotope separation (MLIS) processing of uranium in Japan was initiated by the Institute of Physical and Chemical Research (RIKEN), who achieved a head separation factor as high as 4.7 in a single step using their original process. The Power Reactor and Nuclear Fuel Development Corporation (PNC) has continued the MLIS project in cooperation with RIKEN since 1988. The Japan Atomic Energy Committee will make a decision by about the year 2000 concerning whether or not to carry research and development efforts with respect to the laser enrichment technologies of uranium forward to the next stage. Our MUS development program was previously described in detail. In this paper we briefly discuss the fundamentals of the MLIS process and present some new information about our project.

  7. Phosphorescent organic light emitting diodes with high efficiency and brightness

    DOE Patents [OSTI]

    Forrest, Stephen R; Zhang, Yifan

    2015-11-12

    An organic light emitting device including a) an anode; b) a cathode; and c) an emissive layer disposed between the anode and the cathode, the emissive layer comprising an organic host compound and a phosphorescent compound exhibiting a Stokes Shift overlap greater than 0.3 eV. The organic light emitting device may further include a hole transport layer disposed between the emissive layer and the anode; and an electron transport layer disposed between the emissive layer and the cathode. In some embodiments, the phosphorescent compound exhibits a phosphorescent lifetime of less than 10 .mu.s. In some embodiments, the concentration of the phosphorescent compound ranges from 0.5 wt. % to 10 wt. %.

  8. Atom detection in a two-mode optical cavity with intermediate coupling: Autocorrelation studies

    SciTech Connect (OSTI)

    Norris, D. G.; Cahoon, E. J.; Orozco, L. A.

    2009-10-15

    We use an optical cavity in the regime of intermediate coupling between atom and cavity mode to detect single moving atoms. Degenerate polarization modes allow excitation of the atoms in one mode and collection of spontaneous emission in the other while keeping separate the two sources of light; we obtain a higher confidence and efficiency of detection by adding cavity-enhanced Faraday rotation. Both methods greatly benefit from coincidence detection of photons, attaining fidelities in excess of 99% in less than 1 {mu}s. Detailed studies of the second-order intensity autocorrelation function of light from the signal mode reveal evidence of antibunched photon emissions and the dynamics of single-atom transits.

  9. Luminescent and lasing characteristics of heavily doped Yb{sup 3+}:KY(WO{sub 4}){sub 2} crystals

    SciTech Connect (OSTI)

    Kisel', V E; Troshin, A E; Shcherbitskii, V G; Kuleshov, N V; Pavlyuk, A A; Brunner, F; Paschotta, R; Morier-Genoud, F; Keller, U

    2006-04-30

    The luminescence decay times are measured taking into account reabsorption for KY(WO{sub 4}){sub 2}:Yb(KYW:Yb) crystals with atomic concentrations of active ions from 0.2% to 30%. The radiative lifetime of Yb{sup 3+} ions was measured to be 233 {mu}s. The cw output power of 1.46 and 1.62 W was achieved with the slope efficiency 52% and 47% for Yb:KYW lasers with the atomic concentration of Yb{sup 3+} ions equal to 10% and 30%, respectively. Using a semiconductor mirror with a saturable absorber (SESAM) in the passive mode-locking regime, pulses of duration 194 and 180 fs were obtained at wavelengths of 1042 and 1039 nm for crystals with Yb{sup 3+} concentrations equal to 10% and 30%, respectively, the average output power being 0.63 and 0.75 W. (lasers and amplifiers)

  10. Ion funnel ion trap and process

    DOE Patents [OSTI]

    Belov, Mikhail E [Richland, WA; Ibrahim, Yehia M [Richland, WA; Clowers, Biran H [West Richland, WA; Prior, David C [Hermiston, OR; Smith, Richard D [Richland, WA

    2011-02-15

    An ion funnel trap is described that includes a inlet portion, a trapping portion, and a outlet portion that couples, in normal operation, with an ion funnel. The ion trap operates efficiently at a pressure of .about.1 Torr and provides for: 1) removal of low mass-to-charge (m/z) ion species, 2) ion accumulation efficiency of up to 80%, 3) charge capacity of .about.10,000,000 elementary charges, 4) ion ejection time of 40 to 200 .mu.s, and 5) optimized variable ion accumulation times. Ion accumulation with low concentration peptide mixtures has shown an increase in analyte signal-to-noise ratios (SNR) of a factor of 30, and a greater than 10-fold improvement in SNR for multiply charged analytes.

  11. Critical Magnetic Field Determination of Superconducting Materials

    SciTech Connect (OSTI)

    Canabal, A.; Tajima, T.; Dolgashev, V.A.; Tantawi, S.G.; Yamamoto, T.; /Tsukuba, Natl. Res. Lab. Metrol.

    2011-11-04

    Superconducting RF technology is becoming more and more important. With some recent cavity test results showing close to or even higher than the critical magnetic field of 170-180 mT that had been considered a limit, it is very important to develop a way to correctly measure the critical magnetic field (H{sup RF}{sub c}) of superconductors in the RF regime. Using a 11.4 GHz, 50-MW, <1 {mu}s, pulsed power source and a TE013-like mode copper cavity, we have been measuring critical magnetic fields of superconductors for accelerator cavity applications. This device can eliminate both thermal and field emission effects due to a short pulse and no electric field at the sample surface. A model of the system is presented in this paper along with a discussion of preliminary experimental data.

  12. Regulation of Meiotic Recombination

    SciTech Connect (OSTI)

    Gregory p. Copenhaver

    2011-11-09

    Meiotic recombination results in the heritable rearrangement of DNA, primarily through reciprocal exchange between homologous chromosome or gene conversion. In plants these events are critical for ensuring proper chromosome segregation, facilitating DNA repair and providing a basis for genetic diversity. Understanding this fundamental biological mechanism will directly facilitate trait mapping, conventional plant breeding, and development of genetic engineering techniques that will help support the responsible production and conversion of renewable resources for fuels, chemicals, and the conservation of energy (1-3). Substantial progress has been made in understanding the basal recombination machinery, much of which is conserved in organisms as diverse as yeast, plants and mammals (4, 5). Significantly less is known about the factors that regulate how often and where that basal machinery acts on higher eukaryotic chromosomes. One important mechanism for regulating the frequency and distribution of meiotic recombination is crossover interference - or the ability of one recombination event to influence nearby events. The MUS81 gene is thought to play an important role in regulating the influence of interference on crossing over. The immediate goals of this project are to use reverse genetics to identify mutants in two putative MUS81 homologs in the model plant Arabidopsis thaliana, characterize those mutants and initiate a novel forward genetic screen for additional regulators of meiotic recombination. The long-term goal of the project is to understand how meiotic recombination is regulated in higher eukaryotes with an emphasis on the molecular basis of crossover interference. The ability to monitor recombination in all four meiotic products (tetrad analysis) has been a powerful tool in the arsenal of yeast geneticists. Previously, the qrt mutant of Arabidopsis, which causes the four pollen products of male meiosis to remain attached, was developed as a facile system for assaying recombination using tetrad analysis in a higher eukaryotic system (6). This system enabled the measurement of the frequency and distribution of recombination events at a genome wide level in wild type Arabidopsis (7), construction of genetic linkage maps which include positions for each centromere (8), and modeling of the strength and pattern of interference (9). This proposal extends the use of tetrad analysis in Arabidopsis by using it as the basis for assessing the phenotypes of mutants in genes important for recombination and the regulation of crossover interference and performing a novel genetic screen. In addition to broadening our knowledge of a classic genetic problem - the regulation of recombination by crossover interference - this proposal also provides broader impact by: generating pedagogical tools for use in hands-on classroom experience with genetics, building interdisciplinary collegial partnerships, and creating a platform for participation by junior scientists from underrepresented groups. There are three specific aims: (1) Isolate mutants in Arabidopsis MUS81 homologs using T-DNA and TILLING (2) Characterize recombination levels and interference in mus81 mutants (3) Execute a novel genetic screen, based on tetrad analysis, for genes that regulate meiotic recombination

  13. Modulator considerations for beam chopping in the low energy beam transport at the SSC Laboratory

    SciTech Connect (OSTI)

    Anderson, D.; Pappas, G.

    1991-06-01

    Beam chopping in the low energy transport line at the Superconducting Super Collider Laboratory is accomplished using an electrostatic deflection system. LINAC requirements dictate the design of two modulators operating at 10 Hz with rise and fall times (as measured from approximately 10--99%) of {approximately}100 ns. Design of the first pulser, normally at 10 kV and pulsed to ground potential, utilizes a transformer-coupled diode-clamped solid state circuit to achieve the 2--35 {mu}s pulse width range required. The second pulser, which pulses from ground to approximately 7 kV, relies on a series vacuum tube circuit. The current designs, as well as recent test results and other circuit topologies considered, will be presented. 6 refs.

  14. Study of the fast photoswitching of spin crossover nanoparticles outside and inside their thermal hysteresis loop

    SciTech Connect (OSTI)

    Galle, G.; Degert, J.; Freysz, E.; Etrillard, C.; Letard, J.-F.; Guillaume, F.

    2013-02-11

    We have studied the low spin to high spin phase transition induced by nanosecond laser pulses outside and within the thermal hysteresis loop of the [Fe(Htrz){sub 2} trz](BF{sub 4}){sub 2}-H{sub 2}O spin crossover nanoparticles. We demonstrate that, whatever the temperature of the compound, the photo-switching is achieved in less than 12.5 ns. Outside the hysteresis loop, the photo-induced high spin state remains up to 100 {mu}s and then relaxes. Within the thermal hysteresis loop, the photo-induced high spin state remains as long as the temperature of the sample is kept within the thermal loop. A Raman study indicates that the photo-switching can be completed using single laser pulse excitation.

  15. Emittance Studies of the BNL/SLAC/UCLA 1.6 Cell Photocathode RF Gun

    SciTech Connect (OSTI)

    Palmer, D.T.; Wang, X.J.; Miller, R.H.; Babzien, M.; Ben-Zvi, I.; Pellegrini, C.; Sheehan, J.; Skaritka, J.; Winick, H.; Woodle, M.; Yakimenko, V.; /Brookhaven

    2011-09-09

    The symmetrized 1.6 cell S-band photocathode gun developed by the BNL/SLAC/UCLA collaboration is in operation at the Brookhaven Accelerator Test Facility (ATF). A novel emittance compensation solenoid magnet has also been designed, built and is in operation at the ATF. These two subsystems form an emittance compensated photoinjector used for beam dynamics, advanced acceleration and free electron laser experiments at the ATF. The highest acceleration field achieved on the copper cathode is 150 MV/m, and the guns normal operating field is 130 MV/m. The maximum rf pulse length is 3 {mu}s. The transverse emittance of the photoelectron beam were measured for various injection parameters. The 1 nC emittance results are presented along with electron bunch length measurements that indicated that at above the 400 pC, space charge bunch lengthening is occurring. The thermal emittance, {epsilon}{sub o}, of the copper cathode has been measured.

  16. Possible high power limitations from RF pulsed heating

    SciTech Connect (OSTI)

    Pritzkau, D.P.; Bowden, G.B.; Menegat, A.; Siemann, R.H. [Stanford Linear Accelerator Center, Stanford University, California 94309 (United States)

    1999-05-01

    One of the possible limitations to achieving high power in RF structures is damage to metal surfaces due to RF pulsed heating. Such damage may lead to degradation of RF performance. An experiment to study RF pulsed heating on copper has been developed at SLAC. The experiment consists of operating two pillbox cavities in the TE{sub 011} mode using a 50 MW X-Band klystron. The estimated temperature rise of the surface of copper is 350&hthinsp;{degree}C for a power input of 20 MW to each cavity with a pulse length of 1.5 {mu}s. Preliminary results from an experiment performed earlier are presented. A revised design for continued experiments is also presented along with relevant theory and calculations. {copyright} {ital 1999 American Institute of Physics.}

  17. Aerodynamic force measurement on a large-scale model in a short duration test facility

    SciTech Connect (OSTI)

    Tanno, H.; Kodera, M.; Komuro, T.; Sato, K.; Takahasi, M.; Itoh, K.

    2005-03-01

    A force measurement technique has been developed for large-scale aerodynamic models with a short test time. The technique is based on direct acceleration measurements, with miniature accelerometers mounted on a test model suspended by wires. Measuring acceleration at two different locations, the technique can eliminate oscillations from natural vibration of the model. The technique was used for drag force measurements on a 3 m long supersonic combustor model in the HIEST free-piston driven shock tunnel. A time resolution of 350 {mu}s is guaranteed during measurements, whose resolution is enough for ms order test time in HIEST. To evaluate measurement reliability and accuracy, measured values were compared with results from a three-dimensional Navier-Stokes numerical simulation. The difference between measured values and numerical simulation values was less than 5%. We conclude that this measurement technique is sufficiently reliable for measuring aerodynamic force within test durations of 1 ms.

  18. Suitability of epitaxial GaAs for x-ray imaging

    SciTech Connect (OSTI)

    Sun, G.C.; Talbi, N.; Verdeil, C.; Bourgoin, J.C.

    2004-09-20

    Because the rate of indirect photon-electron conversion for scintillator materials coupled with arrays of photodiodes is at least 25 times smaller than the rate of direct conversion, we examine the conditions to be fulfilled by semiconductors undergoing such direct conversion to be applied to x-ray imaging. Bulk grown materials are not well suited to this application, because large defect concentrations give rise to strongly nonuniform electronic properties. We argue that only epitaxial layers are suitable for use as imaging devices and we illustrate our argument using the case of thick epitaxial GaAs layers. Detectors made with such layers exhibit a good energy resolution, a charge collection efficiency which approaches 1, linearity over more than three orders of amplitude, no afterglow (a response time shorter than 20 {mu}s), and no charge-induced polarization effects.

  19. The earliest events in protein folding: Helix dynamics in proteins and model peptides

    SciTech Connect (OSTI)

    Dyer, R.B.; Williams, S.; Woodruff, W.H. [Los Alamos National Lab., NM (United States)] [and others

    1996-12-31

    The earliest events in protein folding are critically important in determining the folding pathway, but have proved difficult to study by conventional approaches. We have developed new rapid initiation methods and structure-specific probes to interrogate the earliest events of protein folding. Our focus is the pathways. Folding or unfolding reactions are initiated on a fast timescale (10 ns) using a laser induced temperature jump (15 C) and probed with time-resolved infrared spectroscopy. We obtained the kinetics of the helix-coil transition for a model 21-residue peptide. The observed rate constant k{sub obs} = k{sub f} + k{sub u} for reversible kinetics; from the observed rate (6 x 10{sup 6} s{sup -1}) and the equilibrium constant favoring folding of 7.5 at 27 C, we calculate a folding lifetime of 180 ns and an unfolding lifetime of 1.4 {mu}s. The {open_quotes}molten globule{close_quotes} form of apomyoglobin (horse, pH*3, 0.15M NaCl) shows similar kinetics for helix that is unconstrained by tertiary structure (helix with an unusually low Amide I frequency, near 1633 cm{sup -1}). In {open_quotes}native{close_quotes} apomyoglobin (horse, pH*5.3, 10 mM NaCl) two very different rates (45 ns and 70 {mu}s) are observed and we infer that a third occurs on a timescales inaccessible to our experiment (> 1 ms). We suggest that the slower processes are due to helix formation that is rate-limited by the formation of tertiary structure.

  20. SU-E-T-580: Comparison of Cervical Carcinoma IMRT Plans From Four Commercial Treatment Planning Systems (TPS)

    SciTech Connect (OSTI)

    Cao, Y; Li, R; Chi, Z; Zhu, S

    2014-06-01

    Purpose: Different treatment planning systems (TPS) use different treatment optimization and leaf sequencing algorithms. This work compares cervical carcinoma IMRT plans optimized with four commercial TPSs to investigate the plan quality in terms of target conformity and delivery efficiency. Methods: Five cervical carcinoma cases were planned with the Corvus, Monaco, Pinnacle and Xio TPSs by experienced planners using appropriate optimization parameters and dose constraints to meet the clinical acceptance criteria. Plans were normalized for at least 95% of PTV to receive the prescription dose (Dp). Dose-volume histograms and isodose distributions were compared. Other quantities such as Dmin(the minimum dose received by 99% of GTV/PTV), Dmax(the maximum dose received by 1% of GTV/PTV), D100, D95, D90, V110%, V105%, V100% (the volume of GTV/PTV receiving 110%, 105%, 100% of Dp), conformity index(CI), homogeneity index (HI), the volume of receiving 40Gy and 50 Gy to rectum (V40,V50) ; the volume of receiving 30Gy and 50 Gy to bladder (V30,V50) were evaluated. Total segments and MUs were also compared. Results: While all plans meet target dose specifications and normal tissue constraints, the maximum GTVCI of Pinnacle plans was up to 0.74 and the minimum of Corvus plans was only 0.21, these four TPSs PTVCI had significant difference. The GTVHI and PTVHI of Pinnacle plans are all very low and show a very good dose distribution. Corvus plans received the higer dose of normal tissue. The Monaco plans require significantly less segments and MUs to deliver than the other plans. Conclusion: To deliver on a Varian linear-accelerator, the Pinnacle plans show a very good dose distribution. Corvus plans received the higer dose of normal tissue. The Monaco plans have faster beam delivery.

  1. SciSat AM: Stereo 03: Dosmetric evaluation of single versus multi-arc VMAT for lung SBRT

    SciTech Connect (OSTI)

    Karan, T; Taremi, M; Comsa, D; Allibhai, Z; Ryan, M; Le, K

    2014-08-15

    Five non-small cell lung cancer patients previously treated with stereotactic body radiation therapy using the VMAT (volumetric modulated arc therapy) technique were selected for this retrospective study. Plans were re-optimized using Pinnacle treatment planning system (v9.0, Philips Medical), with the basis for comparison a two-arc plan involving a 360 arc in addition to a 90 arc with a couch kick. Additionally a single 360 arc was optimized for comparison, as well as a partial arc covering ?230, avoiding the contralateral lung. All plans met target coverage criteria as dictated by RTOG0236. Plans were evaluated based on conformity, sparing of organs at risk and practical considerations of delivery. Conformity was best in the two-arc plan; however the decrease seen in one- and partial arc plans was not statistically significant as tested by the Wilcoxon rank sum test. The partial-arc plan resulted in the lowest esophagus and trachea dose and the highest heart dose, however none of the plans exceeded organ at risk tolerances for lung SBRT. Partial arcs resulted in plans with slightly cooler dose distributions, a decrease in low dose spillage and an overall lower mean lung dose. The decrease in treatment time was on average 36 and 40 seconds for single and partial arcs, respectively, with partial arcs requiring the lowest number of MUs. The slight decrease in conformity seen in one-arc plans is offset by an increase in efficiency (optimization and treatment time, MUs) making the implementation of a single or partial-arc treatment technique clinically desirable.

  2. SU-E-T-131: Dosimetric Impact and Evaluation of Different Heterogenity Algorithm in Volumetric Modulated Arc Therapy Plan for Stereotactic Ablative Radiotherapy Lung Treatment with the Flattening Filter Free Beam

    SciTech Connect (OSTI)

    Chung, J; Kim, J; Lee, J; Kim, Y

    2014-06-01

    Purpose: The present study aimed to investigate the dosimetric impacts of the anisotropic analytic algorithm (AAA) and the Acuros XB (AXB) plan for lung stereotactic ablative radiation therapy using flattening filter-free (FFF) beam. We retrospectively analyzed 10 patients. Methods: We retrospectively analyzed 10 patients. The dosimetric parameters for the target and organs at risk (OARs) from the treatment plans calculated with these dose calculation algorithms were compared. The technical parameters, such as the computation times and the total monitor units (MUs), were also evaluated. Results: A comparison of DVHs from AXB and AAA showed that the AXB plan produced a high maximum PTV dose by average 4.40% with a statistical significance but slightly lower mean PTV dose by average 5.20% compared to the AAA plans. The maximum dose to the lung was slightly higher in the AXB compared to the AAA. For both algorithms, the values of V5, V10 and V20 for ipsilateral lung were higher in the AXB plan more than those of AAA. However, these parameters for contralateral lung were comparable. The differences of maximum dose for the spinal cord and heart were also small. The computation time of AXB was found fast with the relative difference of 13.7% than those of AAA. The average of monitor units (MUs) for all patients was higher in AXB plans than in the AAA plans. These results indicated that the difference between AXB and AAA are large in heterogeneous region with low density. Conclusion: The AXB provided the advantages such as the accuracy of calculations and the reduction of the computation time in lung stereotactic ablative radiotherapy (SABR) with using FFF beam, especially for VMAT planning. In dose calculation with the media of different density, therefore, the careful attention should be taken regarding the impacts of different heterogeneity correction algorithms. The authors report no conflicts of interest.

  3. TH-E-BRE-05: Analysis of Dosimetric Characteristics in Two Leaf Motion Calculator Algorithms for Sliding Window IMRT

    SciTech Connect (OSTI)

    Wu, L; Huang, B; Rowedder, B; Ma, B; Kuang, Y

    2014-06-15

    Purpose: The Smart leaf motion calculator (SLMC) in Eclipse treatment planning system is an advanced fluence delivery modeling algorithm as it takes into account fine MLC features including inter-leaf leakage, rounded leaf tips, non-uniform leaf thickness, and the spindle cavity etc. In this study, SLMC and traditional Varian LMC (VLMC) algorithms were investigated, for the first time, in dosimetric characteristics and delivery accuracy of sliding window (SW) IMRT. Methods: The SW IMRT plans of 51 cancer cases were included to evaluate dosimetric characteristics and dose delivery accuracy from leaf motion calculated by SLMC and VLMC, respectively. All plans were delivered using a Varian TrueBeam Linac. The DVH and MUs of the plans were analyzed. Three patient specific QA tools - independent dose calculation software IMSure, Delta4 phantom, and EPID portal dosimetry were also used to measure the delivered dose distribution. Results: Significant differences in the MUs were observed between the two LMCs (p≤0.001).Gamma analysis shows an excellent agreement between the planned dose distribution calculated by both LMC algorithms and delivered dose distribution measured by three QA tools in all plans at 3%/3 mm, leading to a mean pass rate exceeding 97%. The mean fraction of pixels with gamma < 1 of SLMC is slightly lower than that of VLMC in the IMSure and Delta4 results, but higher in portal dosimetry (the highest spatial resolution), especially in complex cases such as nasopharynx. Conclusion: The study suggests that the two LMCs generates the similar target coverage and sparing patterns of critical structures. However, SLMC is modestly more accurate than VLMC in modeling advanced MLC features, which may lead to a more accurate dose delivery in SW IMRT. Current clinical QA tools might not be specific enough to differentiate the dosimetric discrepancies at the millimeter level calculated by these two LMC algorithms. NIH/NIGMS grant U54 GM104944, Lincy Endowed Assistant Professorship.

  4. Confinement analyses of the high-density field-reversed configuration plasma in the field-reversed configuration experiment with a liner

    SciTech Connect (OSTI)

    Zhang Shouyin; Intrator, T.P.; Wurden, G.A.; Waganaar, W.J.; Taccetti, J.M.; Renneke, R.; Grabowski, C.; Ruden, E.L.

    2005-05-15

    The focus of the field-reversed configuration (FRC) experiment with a liner (FRX-L) is the formation of a target FRC plasma for magnetized target fusion experiments. An FRC plasma with density of 10{sup 23} m{sup -3}, total temperature in the range of 150-300 eV, and a lifetime of {approx_equal}20 {mu}s is desired. Field-reversed {theta}-pinch technology is used with programed cusp fields at {theta}-coil ends to achieve non-tearing field line reconnections during FRC formation. Well-formed FRCs with density between (2-4)x10{sup 22} m{sup -3}, lifetime in the range of 15-20 {mu}s, and total temperature between 300-500 eV are reproducibly created. Key FRC parameters have standard deviation in the mean of 10% during consecutive shots. The FRCs are formed at 50 mTorr deuterium static fill using 2 kG net reversed bias field inside the {theta}-coil confinement region, with external main field unexpectedly ranging between 15-30 kG. The high-density FRCs confinement properties are approximately in agreement with empirical scaling laws obtained from previous experiments with fill pressure mostly less than 20 mTorr. Analyses in this paper reveal that reducing the external main field modulation and/or extending the {theta}-coil length in the FRX-L device are critical in achieving higher FRC parameters for application in magnetized target fusion.

  5. SU-E-T-495: Neutron Induced Electronics Failure Rate Analysis for a Single Room Proton Accelerator

    SciTech Connect (OSTI)

    Knutson, N; DeWees, T; Klein, E

    2014-06-01

    Purpose: To determine the failure rate as a function of neutron dose of the range modulator's servo motor controller system (SMCS) while shielded with Borated Polyethylene (BPE) and unshielded in a single room proton accelerator. Methods: Two experimental setups were constructed using two servo motor controllers and two motors. Each SMCS was then placed 30 cm from the end of the plugged proton accelerator applicator. The motor was then turned on and observed from outside of the vault while being irradiated to known neutron doses determined from bubble detector measurements. Anytime the motor deviated from the programmed motion a failure was recorded along with the delivered dose. The experiment was repeated using 9 cm of BPE shielding surrounding the SMCS. Results: Ten SMCS failures were recorded in each experiment. The dose per monitor unit for the unshielded SMCS was 0.0211 mSv/MU and 0.0144 mSv/MU for the shielded SMCS. The mean dose to produce a failure for the unshielded SMCS was 63.5 58.3 mSv versus 17.0 12.2 mSv for the shielded. The mean number of MUs between failures were 2297 1891 MU for the unshielded SMCS and 2122 1523 MU for the shielded. A Wilcoxon Signed Ranked test showed the dose between failures were significantly different (P value = 0.044) while the number of MUs between failures were not (P value = 1.000). Statistical analysis determined a SMCS neutron dose of 5.3 mSv produces a 5% chance of failure. Depending on the workload and location of the SMCS, this failure rate could impede clinical workflow. Conclusion: BPE shielding was shown to not reduce the average failure of the SMCS and relocation of the system outside of the accelerator vault was required to lower the failure rate enough to avoid impeding clinical work flow.

  6. JLab, College of W&M researchers study radiation blockers while conducting

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    nuclear imaging of Iodine uptake in mouse tissues | Jefferson Lab JLab, College of W&amp;M researchers study radiation blockers while conducting nuclear imaging of Iodine uptake in mouse tissues JLab, College of W&M researchers study radiation blockers while conducting nuclear imaging of Iodine uptake in mouse tissues April 20, 2005 Scientists have found that a dose five times higher than the FDA-recommended dosage of potassium iodide in the event of a nuclear accident is needed to

  7. I I

    Office of Scientific and Technical Information (OSTI)

    ... (1) as a standard form ODE system y' f(t,y). For the second class, it is possible in theory to ... I mous change in the solution. All of these systems ran caiise problems ...

  8. Monoclonal antibodies to human glycophorin A and cell lines for the production thereof

    DOE Patents [OSTI]

    Vanderlaan, Martin; Bigbee, William L.; Jensen, Ronald H.; Fong, Stella S. N.; Langlois, Richard G.

    1988-01-01

    Cloned mouse hybridoma cell lines have been established which continuously produce antibodies that are highly specific to and exhibit high affinity for glycophorin A.sup.N and differentiate between the M and N forms of human glycophorin A.

  9. Jefferson Lab Detector Technology Aids Development of Cystic...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    the radiotracer, which is trapped in cells containing the transferred gene in the lungs and liver of the mouse. Click image for high-resolution file (7.8MB) Photo credit: Dr....

  10. Our Hidden Past: Biology, part 2 | Y-12 National Security Complex

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    7:11 min. In their new home at "The Mouse House" at Y-12, researchers from ORNL's Biology Division conducted studies that led to standards such as dose rate effects that form...

  11. Crystal structure of the[mu]-opioid receptor bound to a morphinan...

    Office of Scientific and Technical Information (OSTI)

    receptor (mu-OR) in the central nervous system. Here we describe the 2.8 angstrom crystal structure of the mouse mu-OR in complex with an irreversible morphinan antagonist. ...

  12. Structure of Actin Cross-linked with alpha-Actinin: A Network...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    under in vitro conditions. We show in Fig. 1 (left) laser scanning confocal microscope images of the actin cytoskeleton in mouse fibroblast cells. (The image is showing only the...

  13. Regional Dialogue Public Comments & Proposals (contracts/rd)

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    drive, click on the link with your right mouse button and select "Save Target As". Ken Canon, Executive Director, ICNU, June 13, 2005 (PDF, 4 pages, 73 kb, posted June 21, 2005)...

  14. Inhibiting Individual Notch Receptors Improves Treatment

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    in large tumors (approximately 2000 mm3). Bottom: Anti-NRR1 inhibits proper growth of blood vessels, resulting in a "hyper-vascularized" network of blood vessels in mouse neonate...

  15. Los Alamos National Laboratory announces Express Licensing program

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    of the software packages are freely available as either executable downloads or open-source software and may be accessed online with the click of a mouse." "By making access to...

  16. History | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    About the SunShot Initiative » History History This timeline features the key innovations that have advanced the solar industry in the United States. Learn more about these key events from 1955 to present. To see more details, either drag the timeline to the left or right or click the (+) icons. Solar Achievements Timeline INSTRUCTIONS Instructional graphic for mouse use. Click and drag screen or scroll mouse wheel to navigate through timeline. 2011 INDUSTRY Graphic of the U.S. Department of

  17. Annual Report of Site Surveillance and Maintenance Activities at the Rocky Flats Site, Colorado Calendar Year 2015

    Office of Legacy Management (LM)

    7 include annual weed distribution data (for selected species), annual weed control locations, biocontrol release locations, vegetation and wildlife monitoring locations (transect endpoints and sample points), vegetation community classifications, Preble's mouse habitat, wetland locations, wildfire/prescribed burn locations, Preble's mouse and wetland mitigation areas, and rare plant locations. These data are available in various ArcGIS-compatible formats. In addition to these types of spatial

  18. Microsoft Word - S05993_CY2009 Annual Rpt.doc

    Office of Legacy Management (LM)

    6 and sample points), vegetation community classifications, Preble's meadow jumping mouse habitat, wetland locations, wildfire/prescribed burn locations, Preble's meadow jumping mouse and wetland mitigation work, and rare plant locations. These data are available in various ArcGIS ® compatible formats. In addition to these types of spatial data, orthorectified aerial and satellite imagery is also available for the Site for different time frames, including pre- and post-closure. 3.4 Validation

  19. Microsoft Word - S09641_2012Annual.docx

    Office of Legacy Management (LM)

    8 biocontrol release locations, vegetation and wildlife monitoring locations (transect endpoints and sample points), vegetation community classifications, Preble's mouse habitat, wetland locations, wildfire/prescribed burn locations, Preble's mouse and wetland mitigation areas, and rare plant locations. These data are available in various ArcGIS-compatible formats. In addition to these types of spatial data, orthorectified aerial and satellite imagery is also available for the Site for different

  20. Adaptation of existing facilities to isentropic compression experiments

    SciTech Connect (OSTI)

    Tasker, Douglas G; Mielke, Charles H; Rodriguez, George; Rickel, Dwight G

    2011-01-07

    We demonstrate that the established pulsed power infrastructure at the National High Magnetic Field Laboratory - Pulsed Field Facility (NHMFL-PFF) at the Los Alamos National Laboratory can be adapted to obtain high quality isentropic compression experiment (ICE) data on materials in extreme conditions of dynamic high pressure. Experiments utilized a single-turn magnet pulsed power system at the NHMFL-PFF that was originally designed to measure actinide samples in extremes of high magnetic field (to 300 Tesla). A simple modification to the single-turn magnet has converted it to a fast turnaround dynamic high pressure measurement system. This paper details the work done including important background details that indicate that much more can be accomplished with optimization of the load characteristics in terms of ultimate peak pressures. To match the rise time of the NHMFL capacitor bank ({approx}2 {mu}s versus {approx}0.5 {mu}s for the Sandia Z-machine) the sample dimensions can be relatively large, i.e., up to 5 mm thickness. The maximum stresses are {approx}50GPa (0.5 Mbar) at the maximum bank voltage (60 kV) and higher pressures may be possible if the sample is tamped. For the design and predictions of performance of the NHMFL-ICE experiment it is important to have good predictive models. A SPICE code simulation was chosen to model all aspects of the experiment, electrical and physical. To this end, accurate dynamic load models were developed to simulate the compression and expansion of the dynamic load at high pressures using shock physics principles. A series experiments have been performed which demonstrated the feasibility of the NHMFL-ICE technique. The results will be shown and discussed. The NHMFL-ICE technique is an excellent method for measuring equations of state (EOS) at megabar pressures. Because a complete EOS can be obtained in one experiment from zero to the peak pressure, and because many shots can be fired in one day, the technique promises to provide high quality EOS data at relatively low cost.

  1. Knockdown of p53 suppresses Nanog expression in embryonic stem cells

    SciTech Connect (OSTI)

    Abdelalim, Essam Mohamed; Tooyama, Ikuo

    2014-01-10

    Highlights: We investigate the role of p53 in ESCs in the absence of DNA damage. p53 knockdown suppresses ESC proliferation. p53 knockdown downregulates Nanog expression. p53 is essential for mouse ESC self-renewal. -- Abstract: Mouse embryonic stem cells (ESCs) express high levels of cytoplasmic p53. Exposure of mouse ESCs to DNA damage leads to activation of p53, inducing Nanog suppression. In contrast to earlier studies, we recently reported that chemical inhibition of p53 suppresses ESC proliferation. Here, we confirm that p53 signaling is involved in the maintenance of mouse ESC self-renewal. RNA interference-mediated knockdown of p53 induced downregulation of p21 and defects in ESC proliferation. Furthermore, p53 knockdown resulted in a significant downregulation in Nanog expression at 24 and 48 h post-transfection. p53 knockdown also caused a reduction in Oct4 expression at 48 h post-transfection. Conversely, exposure of ESCs to DNA damage caused a higher reduction of Nanog expression in control siRNA-treated cells than in p53 siRNA-treated cells. These data show that in the absence of DNA damage, p53 is required for the maintenance of mouse ESC self-renewal by regulating Nanog expression.

  2. Carcinogenic effects in A/J mice of particulate of a coal-tar paint used in potable water systems

    SciTech Connect (OSTI)

    Robinson, M.; Laurie, R.D.; Bull, R.J.; Stober, J.A.

    1987-01-01

    Coal-tar paints are among the products used as inside coatings for water pipes and storage tanks to retard corrosion in potable water-supply systems. Four different formulations of these paints were tested in earlier work by this laboratory in the Ames mutagenesis and the mouse skin carcinogenesis bioassays(6). The paint most active in these assays was then tested in a particulate form in the lung adenoma assay with A/J mice. The paint was applied to clean glass plates, cured, collected and homogenized in 2% Emulphor. Doses of this coal-tar suspension were administered by gavage at 1.0, 10.0, and 55.0 mg in 0.2 ml per mouse 3 x weekly for 8 weeks. The total doses of coal-tar paint were 24, 240, and 1320 mg/mouse. Benzo(a)pyrene, administered in a parallel schedule to a total dose of 6 mg/mouse, served as positive control. A negative control group received an equivalent volume of 2% Emulphor. Animals were sacrificed at 9 months of age (8 months after first dose) and lung adenomas counted. A dose-related response, in the average number of lung tumors per mouse, was observed with the coal-tar particulate. There were also squamous-cell tumors of the forestomach in 42% of the mice receiving 55.0 mg coal tar paint per application.

  3. Strategies and tools for whole genome alignments

    SciTech Connect (OSTI)

    Couronne, Olivier; Poliakov, Alexander; Bray, Nicolas; Ishkhanov,Tigran; Ryaboy, Dmitriy; Rubin, Edward; Pachter, Lior; Dubchak, Inna

    2002-11-25

    The availability of the assembled mouse genome makespossible, for the first time, an alignment and comparison of two largevertebrate genomes. We have investigated different strategies ofalignment for the subsequent analysis of conservation of genomes that areeffective for different quality assemblies. These strategies were appliedto the comparison of the working draft of the human genome with the MouseGenome Sequencing Consortium assembly, as well as other intermediatemouse assemblies. Our methods are fast and the resulting alignmentsexhibit a high degree of sensitivity, covering more than 90 percent ofknown coding exons in the human genome. We have obtained such coveragewhile preserving specificity. With a view towards the end user, we havedeveloped a suite of tools and websites for automatically aligning, andsubsequently browsing and working with whole genome comparisons. Wedescribe the use of these tools to identify conserved non-coding regionsbetween the human and mouse genomes, some of which have not beenidentified by other methods.

  4. Alkylation damage repair in mammalian genomes

    SciTech Connect (OSTI)

    Mitra, S.; Roy, R.; Kim, N.K. |; Tano, K. |; Ibeanu, G.C. |; Dunn, W.C.; Natarajan, A.T.; Hartenstein, B.; Kaina, B.

    1992-11-01

    The repair of O{sup 6} -alkylguanine in DNA involves only O{sup 6} -methyltransferase (MGMT) while the repair of N-alkylpurines requires multiple proteins including N-methylpurine-DNA glycosylase (MPG). While the biochemical properties human and mouse MGMTs are very similar, the mouse MPG removes 7-methylguanine more efficiently than the human protein. An increased level of MGMT, without a change in the level of MPG associated with gene amplification, was observed in a mouse cell line resistant to 2-chloroethyl-N-nitrosourea. In contrast, no correlation was observed between MPG level and resistance to methyl methanesulfonate in Chinese hamster ovary (CHO) cells. This result suggests a protein other than MPG limits the repair rate of N-alkylpurine in CHO cells.

  5. Alkylation damage repair in mammalian genomes

    SciTech Connect (OSTI)

    Mitra, S.; Roy, R.; Kim, N.K. . Sealy Center for Molecular Science Oak Ridge National Lab., TN ); Tano, K. Oak Ridge National Lab., TN ); Ibeanu, G.C. Oak Ridge National Lab., TN ); Dunn, W.C. (

    1992-01-01

    The repair of O{sup 6} -alkylguanine in DNA involves only O{sup 6} -methyltransferase (MGMT) while the repair of N-alkylpurines requires multiple proteins including N-methylpurine-DNA glycosylase (MPG). While the biochemical properties human and mouse MGMTs are very similar, the mouse MPG removes 7-methylguanine more efficiently than the human protein. An increased level of MGMT, without a change in the level of MPG associated with gene amplification, was observed in a mouse cell line resistant to 2-chloroethyl-N-nitrosourea. In contrast, no correlation was observed between MPG level and resistance to methyl methanesulfonate in Chinese hamster ovary (CHO) cells. This result suggests a protein other than MPG limits the repair rate of N-alkylpurine in CHO cells.

  6. Sources and levels of ambient ocean sound near the antarctic peninsula

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Dziak, Robert P.; Bohnenstiehl, DelWayne R.; Stafford, Kathleen M.; Matsumoto, Haruyoshi; Park, Minkyu; Lee, Won Sang; Fowler, Matt J.; Lau, Tai-Kwan; Haxel, Joseph H.; Mellinger, David K.; et al

    2015-04-14

    Arrays of hydrophones were deployed within the Bransfield Strait and Scotia Sea (Antarctic Peninsula region) from 2005 to 2009 to record ambient ocean sound at frequencies of up to 125 and 500 Hz. Icequakes, which are broadband, short duration signals derived from fracturing of large free-floating icebergs, are a prominent feature of the ocean soundscape. Icequake activity peaks during austral summer and is minimum during winter, likely following freeze-thaw cycles. Iceberg grounding and rapid disintegration also releases significant acoustic energy, equivalent to large-scale geophysical events. Overall ambient sound levels can be as much as ~10–20 dB higher in the open,more » deep ocean of the Scotia Sea compared to the relatively shallow Bransfield Strait. Noise levels become lowest during the austral winter, as sea-ice cover suppresses wind and wave noise. Ambient noise levels are highest during austral spring and summer, as surface noise, ice cracking and biological activity intensifies. Vocalizations of blue (Balaenoptera musculus) and fin (B. physalus) whales also dominate the long-term spectra records in the 15–28 and 89 Hz bands. Blue whale call energy is a maximum during austral summer-fall in the Drake Passage and Bransfield Strait when ambient noise levels are a maximum and sea-ice cover is a minimum. Fin whale vocalizations were also most common during austral summer-early fall months in both the Bransfield Strait and Scotia Sea. The hydrophone data overall do not show sustained anthropogenic sources (ships and airguns), likely due to low coastal traffic and the typically rough weather and sea conditions of the Southern Ocean.« less

  7. Deflagration-to-detonation transition project: quarterly report for the period September through November 1979

    SciTech Connect (OSTI)

    Lieberman, M. L.

    1980-07-01

    The activities of the Sandia Laboratories project on deflagration-to-detonation transition (DDT) pertain primarily to the development of small, safe, low-voltage, hot-wire detonators. Its major goals are: the formulation of a modeling capability for DDT of the explosive 2-(5-cyanotetrazolato)pentaamminecobalt(III) perchlorate (CP); the development of improved DDT materials; the establishment of a data base for corrosion, compatibility, and reliability of CP-loaded detonators; and the design and development of advanced DDT components. Progress in this research is reported. The planned development of the MC3423 detonator has been completed and the final design review meeting has been held. Additional work must be performed to establish satisfactory output function. Ignition sensitivity data have also been obtained. Ignition and shock testing experiments for development of the MC3533 detonator have been planned. An initial version of the component will utilize available MC3423 headers, while the final design will incorporate a new header that has been designed and ordered. Detonator performance studies have been planned to optimize CP density-length factors. Feasibility studies on the MC3196A detonator have continued in an effort to obtain a reliable 50-200 ..mu..s function time.

  8. Characteristics of high-rate energy spectroscopy systems using HPGe coaxial detectors and time-variant filters

    SciTech Connect (OSTI)

    Britton, C.L.; Becker, T.H.; Paulus, T.J.; Trammell, R.C.

    1984-02-01

    A high-rate, high-resolution gamma spectrometer system is described. The system consists of a reverse electrode HPGe coaxial detector, a transistor reset preamplifier, an active, semi-Gaussian prefilter, a gated integrator, and a unique data acquisition system consisting of a 10 ..mu..s, 13 bit ADC, fast FIFO memory, 8k by 23 bit data memory, and computer interface circuitry under the control of a Z-80A ..mu..P. The effects of the various components on the throughput are described and throughput data is presented. The resolution and peak shift for various shaping times are presented for count rates up to 1 Mcps input rate using a mixed /sup 22/Na and /sup 60/Co source. The low rate resolutions of /sup 57/Co and /sup 60/Co for various shaping times using either the semi-Gaussian or gated integrator output are discussed as well as the low energy resolution and peak shifts in the presence of high energy events.

  9. GTA Beamloss-Monitor System

    SciTech Connect (OSTI)

    Rose, C.R.; Fortgang, C.M.; Power, J.P.

    1992-01-01

    The GTA Beamless-Monitor System at Los Alamos National Laboratory has been designed to detect high-energy particle loss in the accelerator beamline and shut down the accelerator before any damage can occur. To do this, the Beamless-Monitor System measures the induced gamma radiation, from (p, {gamma}) reactions, at 15 selected points along the beamline, converts this measured radiation to electrical signals integrates and compares them to preset limits, and, in the event of an over-limit condition causes the Fast-Protect System to shut down the entire accelerator. The system dynamic range exceeds 70 dB which will enable experimenters to use the Beamless-Monitor System to help steer the beam as well as provide signals for a Fast-Protect System. The system response time is less than 7 {mu}s assuming a step-function, worst-case beam spill of 50 mA. The system resolution, based on the noise floor of the electronics is about 1.3 mRads/s. Production units have been built and meet the above specifications. The remainder of the system will be installed and tested later in 1992/1993 with the GTA accelerator. The ionization chamber sensitivity and response time are described in the paper.

  10. GTA Beamloss-Monitor System

    SciTech Connect (OSTI)

    Rose, C.R.; Fortgang, C.M.; Power, J.P.

    1992-09-01

    The GTA Beamless-Monitor System at Los Alamos National Laboratory has been designed to detect high-energy particle loss in the accelerator beamline and shut down the accelerator before any damage can occur. To do this, the Beamless-Monitor System measures the induced gamma radiation, from (p, {gamma}) reactions, at 15 selected points along the beamline, converts this measured radiation to electrical signals integrates and compares them to preset limits, and, in the event of an over-limit condition causes the Fast-Protect System to shut down the entire accelerator. The system dynamic range exceeds 70 dB which will enable experimenters to use the Beamless-Monitor System to help steer the beam as well as provide signals for a Fast-Protect System. The system response time is less than 7 {mu}s assuming a step-function, worst-case beam spill of 50 mA. The system resolution, based on the noise floor of the electronics is about 1.3 mRads/s. Production units have been built and meet the above specifications. The remainder of the system will be installed and tested later in 1992/1993 with the GTA accelerator. The ionization chamber sensitivity and response time are described in the paper.

  11. Optical absorption and stimulated emission of neodymium in yttrium lithium fluoride

    SciTech Connect (OSTI)

    Ryan, J.R.; Beach, R. )

    1992-10-01

    A spectroscopic investigation of Nd{sup 3+} in yttrium lithium fluoride was performed. Spectrally and orientationally resolved cross sections for the {sup 4}{ital F}{sub 3/2}--{sup 4}{ital I}{sub 11/2} and {sup 4}{ital F}{sub 3/2}--{sup 4}{ital I}{sub 9/2} transitions are presented. We applied the Judd--Ofelt theory to measured absorption spectra to determine the orientation-averaged intensity parameters {Omega}{sub 2}=0.362{times}10{sup {minus}20} cm{sup 2}, {Omega}{sub 4}=4.02{times}10{sup {minus}20} cm{sup 2}, and {Omega}{sub 6}=4.84{times}10{sup {minus}20} cm{sup 2}. Using these intensity parameters, we predicted the radiative lifetime of the metastable {sup 4}{ital F}{sub 3/2} state to be 525 {mu}s, in excellent agreement with measured {sup 4}{ital F}{sub 3/2} decay signatures. Finally, absorption cross-section data are presented that will be of interest to the laser designer.

  12. Experimental study of a simple method to chop Penning SPS H{sup {minus}} beams

    SciTech Connect (OSTI)

    Smith, H.V. Jr.; Allison, P.; Schneider, J.D.; Stelzer, J.E.

    1994-08-01

    Accumulator rings proposed for use in high-intensity spallation-neutron sources require a chopped beam with particle-free gaps {approx}100 ns wide at 1--2 MHz rates with rise and fall times {le} 20 ns. Chopping the beam directly in the ion source may be an attractive way to provide the desired beam structure. A grounded collar placed in the drift region next to the emission aperture lowers the e{sup {minus}}/H{sup {minus}} ratio in the 8X source H{sup {minus}} beam. We electrically isolated the collar and biased it to modulate the extracted H{sup {minus}} current. Positive collar bias decreases the H{sup {minus}} beam by up to 90%. The fastest H{sup {minus}} current fall and rise times achieved to date are 400 ns and 2 {mu}s, respectively. The fall time is close to the pulser rise time ({approx}300 ns). The rise time is considerably longer than the pulser fall time ({approx}500 ns). Negative collar bias lowers the H{sup {minus}} beam by up to 50%.

  13. Propagation of gamma rays and production of free electrons in air

    SciTech Connect (OSTI)

    Dimant, Y. S.; Nusinovich, G. S.; Romero-Talamas, C. A.; Granatstein, V. L.; Sprangle, P.; Penano, J.

    2012-10-15

    This paper is devoted to the analysis of production of free electrons in air by gamma-rays leaking from radioactive materials. A model based on the Klein-Nishina scattering theory is used to calculate scattering cross sections and approximate the electron production rate. The model includes the effects of primary gamma-quanta radiated by the source as well as that scattered in air. Comparison of the model with the mcnpx kinetic code (http://mcnpx.lanl.gov/) in a sample problem shows excellent agreement. The motivation for this research comes from the recently proposed concept of remote detection of concealed radioactive materials [V. L. Granatstein and G. S. Nusinovich, J. Appl. Phys. 108, 063304 (2010)]. The concept is based on the breakdown in air at the focal point of a high-power beam of electromagnetic waves produced by a THz gyrotron with a 10-20 {mu}s pulse. The presence of a radioactive material can greatly exceed the production rate of free electrons over the natural background rate. Additional electrons act as seeds to initiate the breakdown and create sufficiently dense plasma at the focal region. The dense plasma can then be remotely detected as an unambiguous effect of the concealed radioactive material.

  14. Spectrographic temperature measurement of a high power breakdown arc in a high pressure gas switch

    SciTech Connect (OSTI)

    Yeckel, Christopher; Curry, Randy

    2011-09-15

    A procedure for obtaining an approximate temperature value of conducting plasma generated during self-break closure of a RIMFIRE gas switch is described. The plasma is in the form of a breakdown arc which conducts approximately 12 kJ of energy in 1 {mu}s. A spectrographic analysis of the trigger-section of the 6-MV RIMFIRE laser triggered gas switch used in Sandia National Laboratory's ''Z-Machine'' has been made. It is assumed that the breakdown plasma has sufficiently approached local thermodynamic equilibrium allowing a black-body temperature model to be applied. This model allows the plasma temperature and radiated power to be approximated. The gas dielectric used in these tests was pressurized SF{sub 6}. The electrode gap is set at 4.59 cm for each test. The electrode material is stainless steel and insulator material is poly(methyl methacrylate). A spectrum range from 220 to 550 nanometers has been observed and calibrated using two spectral irradiance lamps and three spectrograph gratings. The approximate plasma temperature is reported.

  15. An innovative high-power constant-current pulsed-arc power-supply for a high-density pulsed-arc-plasma ion-source using a LaB{sub 6}-filament

    SciTech Connect (OSTI)

    Ueno, A.; Oguri, H.; Ikegami, K.; Namekawa, Y.; Ohkoshi, K.; Tokuchi, A.

    2010-02-15

    An innovative high-power constant-current (CC) pulsed-arc (PA) power-supply (PS) indispensable for a high-density PA plasma ion-source using a lanthanum hexaboride (LaB{sub 6}) filament was devised by combining a constant-voltage (CV) PA-PS, which is composed of an insulated gate bipolar transistor (IGBT) switch, a CV direct-current (dc) PS and a 270 mF capacitor with a CC-PA-PS, which is composed of an IGBT-switch, a CC-dc-PS and a 400 {mu}H inductor, through the inductor. The hybrid-CC-PA-PS succeeded in producing a flat arc-pulse with a peak power of 56 kW (400 Ax140 V) and a duty factor of more than 1.5%(600 {mu}sx25 Hz) for Japan Proton Accelerator Research Complex (J-PARC) H{sup -} ion-source stably. It also succeeded in shortening the 99% rising-time of the arc-pulse-current to about 20 {mu}s and tilting up or down the arc-pulse-current arbitrarily and almost linearly by changing the setting voltage of its CV-dc-PS.

  16. Generation of high charge state metal ion beams by electron cyclotron resonance heating of vacuum arc plasma in cusp trap

    SciTech Connect (OSTI)

    Nikolaev, A. G.; Savkin, K. P.; Oks, E. M.; Vizir, A. V.; Yushkov, G. Yu.; Vodopyanov, A. V.; Izotov, I. V.; Mansfeld, D. A.

    2012-02-15

    A method for generating high charge state heavy metal ion beams based on high power microwave heating of vacuum arc plasma confined in a magnetic trap under electron cyclotron resonance conditions has been developed. A feature of the work described here is the use of a cusp magnetic field with inherent ''minimum-B'' structure as the confinement geometry, as opposed to a simple mirror device as we have reported on previously. The cusp configuration has been successfully used for microwave heating of gas discharge plasma and extraction from the plasma of highly charged, high current, gaseous ion beams. Now we use the trap for heavy metal ion beam generation. Two different approaches were used for injecting the vacuum arc metal plasma into the trap - axial injection from a miniature arc source located on-axis near the microwave window, and radial injection from sources mounted radially at the midplane of the trap. Here, we describe preliminary results of heating vacuum arc plasma in a cusp magnetic trap by pulsed (400 {mu}s) high power (up to 100 kW) microwave radiation at 37.5 GHz for the generation of highly charged heavy metal ion beams.

  17. Charge-state-resolved ion energy distribution functions of cathodic vacuum arcs: A study involving the plasma potential and biased plasmas

    SciTech Connect (OSTI)

    Anders, Andre; Oks, Efim

    2007-02-15

    Charge-state-resolved ion energy distribution functions were measured for pulsed cathodic arcs taking the sheath into account that formed between the plasma and the entrance of a combined energy and mass spectrometer. An electron emitting probe was employed to independently determine the plasma potential. All results were obtained by averaging over several individual measurements because the instantaneous energy distributions and the plasma potential show large amplitude fluctuations due to the explosive nature of the arc plasma generation. It was found that the ion energy distribution functions in the plasma were independent of the ion charge state. This is in contrast to findings with continuously operating, direct-current arcs that employ a magnetic field at the cathode to steer the cathode spot motion. The different findings indicate the important role of the magnetic steering field for the plasma properties of direct-current arcs. The results are further supported by experiments with 'biased plasmas' obtained by shifting the potential of the anode. Finally, it was shown that the ion energy distributions were broader and shifted to higher energy at the beginning of each arc pulse. The characteristic time for relaxation to steady state distributions is about 100 {mu}s.

  18. High current multicharged metal ion source using high power gyrotron heating of vacuum arc plasma

    SciTech Connect (OSTI)

    Vodopyanov, A. V.; Golubev, S. V.; Khizhnyak, V. I.; Mansfeld, D. A.; Nikolaev, A. G.; Oks, E. M.; Savkin, K. P.; Vizir, A. V.; Yushkov, G. Yu.

    2008-02-15

    A high current, multi charged, metal ion source using electron heating of vacuum arc plasma by high power gyrotron radiation has been developed. The plasma is confined in a simple mirror trap with peak magnetic field in the plug up to 2.5 T, mirror ratio of 3-5, and length variable from 15 to 20 cm. Plasma formed by a cathodic vacuum arc is injected into the trap either (i) axially using a compact vacuum arc plasma gun located on axis outside the mirror trap region or (ii) radially using four plasma guns surrounding the trap at midplane. Microwave heating of the mirror-confined, vacuum arc plasma is accomplished by gyrotron microwave radiation of frequency 75 GHz, power up to 200 kW, and pulse duration up to 150 {mu}s, leading to additional stripping of metal ions by electron impact. Pulsed beams of platinum ions with charge state up to 10+, a mean charge state over 6+, and total (all charge states) beam current of a few hundred milliamperes have been formed.

  19. Characterization of Arc Generated Plasma Interactions with a Liquid Metal Medium

    SciTech Connect (OSTI)

    Hahn, Gregory C.; Martin, Elijah H.; Bourham, Mohamed A.

    2005-05-15

    Plasma interaction with first wall and interior reactor chamber components is an influencing factor in the design of inertial fusion facilities. The concept of a liquid metal wall, in which a circulating lithium curtain would be used, has been considered in many studies. The interaction of plasmas with moving liquid metals is a complex subject due to the influence of hydrodynamics, evaporation and droplet formation, nucleation and agglomeration of condensed particulates. To gain an understanding of some of the specific details of this interaction an experimental setup of an arc-generated plasma interacting with a liquid lead pool has been designed, constructed and operated. This simulation of the plasma-liquid interaction focuses on the particle condensation of the liquid metal after plasma interaction. The experiment generates transient high-density plasma over 50 {mu}s pulse duration. Plasma characteristics are determined by various diagnostics. A set of collection substrates are arranged to collect nucleated particulates condensing from the evolving plume. Particulate size and distribution are analyzed numerically using digital images.

  20. Nature of high-energy ions in the cathode plasma jet of a vacuum arc with high rate of current rise

    SciTech Connect (OSTI)

    Beilis, I.I.

    2004-10-04

    The production mechanism of extremely high-energy (up to 10 keV) ions observed in vacuum arcs having only a few tens of volts of arc voltage was considered. A model was developed for the plasma acceleration in a high-current ({>=}1 kA) short pulsed (<1 {mu}s) vacuum arc, taking into account the high rate of rise of the spot current (dI/dt>100 MA/s). A system of equations, including equations for the cathode spot and the plasma jet, was solved self-consistently with dI/dt in the range of 0.1-10 GA/s. It was shown that the plasma could be accelerated to the measured energy in the near spot region due to a gas dynamic mechanism and that the ion energy depends on the ratio of the ion flux to the electron flux. This ratio is determined by the cathode erosion rate. The calculated cathode erosion rate varies from 200 to 10 {mu}g/C when the ion energy increases from 0.1 to 10 keV and well agrees with measurements.

  1. Dedicated Laboratory Setup for CO{sub 2} TEA Laser Propulsion Experiments at Rensselaer Polytechnic Institute

    SciTech Connect (OSTI)

    Salvador, Israel I.; Kenoyer, David; Myrabo, Leik N.; Notaro, Samuel

    2010-10-08

    Laser propulsion research progress has traditionally been hindered by the scarcity of photon sources with desirable characteristics, as well as integrated specialized flow facilities in a dedicated laboratory environment. For TEA CO{sub 2} lasers, the minimal requirements are time-average powers of >100 W), and pulse energies of >10 J pulses with short duration (e.g., 0.1 to 1 {mu}s); furthermore, for the advanced pulsejet engines of interest here, the laser system must simulate pulse repetition frequencies of 1-10 kilohertz or more, at least for two (carefully sequenced) pulses. A well-equipped laser propulsion laboratory should have an arsenal of sensor and diagnostics tools (such as load cells, thrust stands, moment balances, pressure and heat transfer gages), Tesla-level electromagnet and permanent magnets, flow simulation facilities, and high-speed visualization systems, in addition to other related equipment, such as optics and gas supply systems. In this paper we introduce a cutting-edge Laser Propulsion Laboratory created at Rensselaer Polytechnic Institute, one of the very few in the world to be uniquely set up for beamed energy propulsion (BEP) experiments. The present BEP research program is described, along with the envisioned research strategy that will exploit current and expanded facilities in the near future.

  2. Electric field induced needle-pulsed arc discharge carbon nanotube production apparatus: Circuitry and mechanical design

    SciTech Connect (OSTI)

    Kia, Kaveh Kazemi; Bonabi, Fahimeh

    2012-12-15

    A simple and low cost apparatus is reported to produce multiwall carbon nanotubes and carbon nano-onions by a low power short pulsed arc discharge reactor. The electric circuitry and the mechanical design details and a micro-filtering assembly are described. The pulsed-plasma is generated and applied between two graphite electrodes. The pulse width is 0.3 {mu}s. A strong dc electric field is established along side the electrodes. The repetitive discharges occur in less than 1 mm distance between a sharp tip graphite rod as anode, and a tubular graphite as cathode. A hydrocarbon vapor, as carbon source, is introduced through the graphite nozzle in the cathode assembly. The pressure of the chamber is controlled by a vacuum pump. A magnetic field, perpendicular to the plasma path, is provided. The results show that the synergetic use of a pulsed-current and a dc power supply enables us to synthesize carbon nanoparticles with short pulsed plasma. The simplicity and inexpensiveness of this plan is noticeable. Pulsed nature of plasma provides some extra degrees of freedom that make the production more controllable. Effects of some design parameters such as electric field, pulse frequency, and cathode shape are discussed. The products are examined using scanning probe microscopy techniques.

  3. Neutron measurements in search of cold fusion

    SciTech Connect (OSTI)

    Anderson, R.E.; Goulding, C.A.; Johnson, M.W.; Butterfield, K.B.; Gottesfeld, S.; Baker, D.A.; Springer, T.E.; Garzon, F.H.; Bolton, R.D.; Leonard, E.M.; Chancellor, T. )

    1991-05-10

    We have conducted a search for neutron emission from cold fusion systems of the electrochemical type and, to a lesser extent, the high-pressure gas cell type. Using a high-efficiency well counter and an NE 213 scintillator, the experiments were conducted on the earth's surface and in a shielded cave approximately 50 ft underground. After approximately 6500 h of counting time, we have obtained no evidence for cold fusion processes leading to neutron production. However, we have observed all three types of neutron data that have been presented as evidence for cold fusion: large positive fluctuations in the neutron counting rate, weak peaks near 2.5 MeV in the neutron energy spectrum, and bursts of up to 140 neutrons in 500-{mu}s intervals. The data were obtained under circumstances that clearly show our results to be data encountered as a part of the naturally occurring neutron background, which is due primarily to cosmic rays. Thus, observing these types of data does not, of itself, provide evidence for the existence of cold fusion processes. Artifacts in the data that were due to counter misbehavior were also observed to lead to long-term neutron bursts'' whose time duration varied from several hours to several days. We conclude that any experiments which attempt to observed neutron emission must include strong steps to ensure that the experiments deal adequately with both cosmic-ray processes and counter misbehavior.

  4. Progress on the Los Alamos heavy-ion injector

    SciTech Connect (OSTI)

    Wilson, D.C.; Riepe, K.B.; Ballard, E.O.; Meyer, E.A.; Shurter, R.P.; Van Haaften, F.W.; Humphries, S. Jr.

    1986-01-01

    Heavy-ion fusion using an induction linac requires injection of multiple high-current beams from a pulsed electrostatic accelerator at as high a voltage as practical. Los Alamos National Laboratory is developing a 16-beam, 2-MeV, pulsed electrostatic accelerator for Al/sup +/ ions. The ion source will use a pulsed metal vapor arc plasma. A biased grid will control plasma flux into the ion extraction region. This source has achieved a normalized emittance of epsilon/sub n/ < 3.10/sup -7/..pi..-m-rad with Al/sup +/ ions. An 800 kV Marx prototype with a laser fired diverter is being assembled. The ceramic accelerating column sections have been brazed and leak tested. Voltage hold off on a brazed sample was more than doubled by selective removal of the Ticusil braze fillet extending along the ceramic. A scaled test module held 250 kV for 50 ..mu..s, giving confidence that the full module can hold 175 kV per section. The pressure vessel should be received in June 1986. High-voltage testing of a 1 MV column will begin by early 1987.

  5. Ultrafast studies of solution dynamics

    SciTech Connect (OSTI)

    Woodruff, W.H.; Dyer, R.B.; Callender, R.H.

    1997-10-01

    This is the final report of a one-year, Laboratory Directed Research and Development (LDRD) project at Los Alamos National Laboratory (LANL). Fast chemical dynamics generally must be initiated photochemically. This limits the applicability of modern laser methods for following the structural changes that occur during chemical and biological reactions to those systems that have an electronic chromophore that has a significant yield of photoproduct when excited. This project has developed a new and entirely general approach to ultrafast initiation of reactions in solution: laser-induced temperature jump (T-jump). The results open entire new fields of study of ultrafast molecular dynamics in solution. The authors have demonstrated the T-jump technique on time scales of 50 ps and longer, and have applied it to study of the fast events in protein folding. They find that a general lifetime of alpha-helix formation is ca 100 ns, and that tertiary folds (in apomyoglobin) form in ca 100 {mu}s.

  6. Design of an ultra low power CMOS pixel sensor for a future neutron personal dosimeter

    SciTech Connect (OSTI)

    Zhang, Y.; Hu-Guo, C.; Husson, D.; Hu, Y.

    2011-07-01

    Despite a continuously increasing demand, neutron electronic personal dosimeters (EPDs) are still far from being completely established because their development is a very difficult task. A low-noise, ultra low power consumption CMOS pixel sensor for a future neutron personal dosimeter has been implemented in a 0.35 {mu}m CMOS technology. The prototype is composed of a pixel array for detection of charged particles, and the readout electronics is integrated on the same substrate for signal processing. The excess electrons generated by an impinging particle are collected by the pixel array. The charge collection time and the efficiency are the crucial points of a CMOS detector. The 3-D device simulations using the commercially available Synopsys-SENTAURUS package address the detailed charge collection process. Within a time of 1.9 {mu}s, about 59% electrons created by the impact particle are collected in a cluster of 4 x 4 pixels with the pixel pitch of 80 {mu}m. A charge sensitive preamplifier (CSA) and a shaper are employed in the frond-end readout. The tests with electrical signals indicate that our prototype with a total active area of 2.56 x 2.56 mm{sup 2} performs an equivalent noise charge (ENC) of less than 400 e - and 314 {mu}W power consumption, leading to a promising prototype. (authors)

  7. Half-life measurements of isomeric states populated in projectile fragmentation

    SciTech Connect (OSTI)

    Bowry, M.; Podolay, Zs.

    2012-10-20

    The half-lives of excited isomeric states observed in {sup 195}Au, {sup 201}Tl and {sup 215}Rn are reported for the first time. Delayed {gamma}-rays were correlated with nuclei produced in the projectile fragmentation of relativistic {sup 238}U ions, unambiguously identified in terms of their atomic number (Z) and mass-to-charge ratio (A/Q) after traversing an in-flight separator. The observation of a long-lived isomeric state in {sup 195}Au with t{sub 1/2} = 16{sub -4}{sup +8}{mu}s is presented. Two shorter-lived isomeric states were detected in {sup 201}Tl and {sup 215}Rn with t{sub 1/2} = 95{sub -21}{sup +39} and 57{sub -12}{sup +21} ns respectively. In total 24 isomeric states were identified in different nuclei from Pt to Rn (A {approx} 200) during the current study, the majority of which were previously reported. The wealth of spectroscopic data provides the opportunity to determine the isomeric ratios over a wide range of Z, A and angular momentum (I h) of the reaction products. In particular, high-spin states with I Greater-Than-Or-Equivalent-To 18 h provide a robust test of theoretical models of fragmentation.

  8. Is the resilience of C{sub 60}{sup +} towards decomposition a question of time?

    SciTech Connect (OSTI)

    Lifshitz, C.; Sandler, P.; Gotkis, I.; Laskin, J.

    1993-12-31

    The authors have measured kinetic energy release distributions (KERDs) for the evaporation of C{sub 2} from C{sub 60}{sup +} and other fullerene cations. Through analysis of the KERDs by Klot`s theory the authors determined binding energies -- the C{sub 2} binding energy in C{sub 60}{sup +} is 5.23 eV. A thermochemical cycle gave 5.77 eV as the C{sub 2} binding energy in C{sub 60} neutral. Once these binding energies were known, it enabled calculations of time resolved breakdown curves for parent and daughter ions. These calculations included the option of energy relaxation through emission in the infra-red. It was found that C{sub 60}{sup +} is almost completely undissociated at t = 1 {mu}s at an internal energy of 30 eV. However, at t = 1s, it is completely dissociated. An intrinsic shift (IS) of 16.2 eV is predicted. These results explain SID (Surface Induced Decomposition) data by Whetten and coworkers. The applicability of Trouton`s Rule and evaporation to fullerene decompositions will be discussed. Alternative mechanisms, including the Rice {open_quotes}shrink wrap{close_quotes} mechanism, will be discussed in view of the apparent universality of the Gspann parameter, {gamma}=23.5.

  9. Intensity-modulated radiosurgery with rapidarc for multiple brain metastases and comparison with static approach

    SciTech Connect (OSTI)

    Wang Jiazhu; Pawlicki, Todd; Rice, Roger; Mundt, Arno J.; Sandhu, Ajay; Lawson, Joshua; Murphy, Kevin T.

    2012-04-01

    Rotational RapidArc (RA) and static intensity-modulated radiosurgery (IMRS) have been used for brain radiosurgery. This study compares the 2 techniques from beam delivery parameters and dosimetry aspects for multiple brain metastases. Twelve patients with 2-12 brain lesions treated with IMRS were replanned using RA. For each patient, an optimal 2-arc RA plan from several trials was chosen for comparison with IMRS. Homogeneity, conformity, and gradient indexes have been calculated. The mean dose to normal brain and maximal dose to other critical organs were evaluated. It was found that monitor unit (MU) reduction by RA is more pronounced for cases with larger number of brain lesions. The MU-ratio of RA and IMRS is reduced from 104% to 39% when lesions increase from 2 to 12. The dose homogeneities are comparable in both techniques and the conformity and gradient indexes and critical organ doses are higher in RA. Treatment time is greatly reduced by RA in intracranial radiosurgery, because RA uses fewer MUs, fewer beams, and fewer couch angles.

  10. X-ray diffraction in the pulsed laser heated diamond anvil cell

    SciTech Connect (OSTI)

    Goncharov, Alexander F.; Struzhkin, Viktor V.; Dalton, D. Allen; Prakapenka, Vitali B.; Kantor, Innokenty; Rivers, Mark L.

    2010-11-15

    We have developed in situ x-ray synchrotron diffraction measurements of samples heated by a pulsed laser in the diamond anvil cell at pressure up to 60 GPa. We used an electronically modulated 2-10 kHz repetition rate, 1064-1075 nm fiber laser with 1-100 {mu}s pulse width synchronized with a gated x-ray detector (Pilatus) and time-resolved radiometric temperature measurements. This enables the time domain measurements as a function of temperature in a microsecond time scale (averaged over many events, typically more than 10 000). X-ray diffraction data, temperature measurements, and finite element calculations with realistic geometric and thermochemical parameters show that in the present experimental configuration, samples 4 {mu}m thick can be continuously temperature monitored (up to 3000 K in our experiments) with the same level of axial and radial temperature uniformities as with continuous heating. We find that this novel technique offers a new and convenient way of fine tuning the maximum sample temperature by changing the pulse width of the laser. This delicate control, which may also prevent chemical reactivity and diffusion, enables accurate measurement of melting curves, phase changes, and thermal equations of state.

  11. Dynamic processes in field-reversed-configuration compact toroids

    SciTech Connect (OSTI)

    Rej, D.J.

    1987-01-01

    In this lecture, the dynamic processes involved in field-reversed configuration (FRC) formation, translation, and compression will be reviewed. Though the FRC is related to the field-reversed mirror concept, the formation method used in most experiments is a variant of the field-reversed THETA-pinch. Formation of the FRC eqilibrium occurs rapidly, usually in less than 20 ..mu..s. The formation sequence consists of several coupled processes: preionization; radial implosion and compression; magnetic field line closure; axial contraction; equilibrium formation. Recent experiments and theory have led to a significantly improved understanding of these processes; however, the experimental method still relies on a somewhat empirical approach which involves the optimization of initial preionization plasma parameters and symmetry. New improvements in FRC formation methods include the use of lower voltages which extrapolate better to larger devices. The axial translation of compact toroid plasmas offers an attractive engineering convenience in a fusion reactor. FRC translation has been demonstrated in several experiments worldwide, and these plasmas are found to be robust, moving at speeds up to the Alfven velocity over distances of up to 16 m, with no degradation in the confinement. Compact toroids are ideal for magnetic compression. Translated FRCs have been compressed and heated by imploding liners. Upcoming experiments will rely on external flux compression to heat a translater FRC at 1-GW power levels. 39 refs.

  12. High speed curved position sensitive detector

    DOE Patents [OSTI]

    Hendricks, Robert W.; Wilson, Jack W.

    1989-01-01

    A high speed curved position sensitive porportional counter detector for use in x-ray diffraction, the detection of 5-20 keV photons and the like. The detector employs a planar anode assembly of a plurality of parallel metallic wires. This anode assembly is supported between two cathode planes, with at least one of these cathode planes having a serpentine resistive path in the form of a meander having legs generally perpendicular to the anode wires. This meander is produced by special microelectronic fabrication techniques whereby the meander "wire" fans outwardly at the cathode ends to produce the curved aspect of the detector, and the legs of the meander are small in cross-section and very closely spaced whereby a spatial resolution of about 50 .mu.m can be achieved. All of the other performance characteristics are about as good or better than conventional position sensitive proportional counter type detectors. Count rates of up to 40,000 counts per second with 0.5 .mu.s shaping time constants are achieved.

  13. Binding Preferences, Surface Attachment, Diffusivity, and Orientation of a Family 1 Carbohydrate-Binding Module on Cellulose

    SciTech Connect (OSTI)

    Nimlos, M. R.; Beckham, G. T.; Matthews, J. F.; Bu, L.; Himmel, M. E.; Crowley, M. F.

    2012-06-08

    Cellulase enzymes often contain carbohydrate-binding modules (CBMs) for binding to cellulose. The mechanisms by which CBMs recognize specific surfaces of cellulose and aid in deconstruction are essential to understand cellulase action. The Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase, Cel7A, is known to selectively bind to hydrophobic surfaces of native cellulose. It is most commonly suggested that three aromatic residues identify the planar binding face of this CBM, but several recent studies have challenged this hypothesis. Here, we use molecular simulation to study the CBM binding orientation and affinity on hydrophilic and hydrophobic cellulose surfaces. Roughly 43 {mu}s of molecular dynamics simulations were conducted, which enables statistically significant observations. We quantify the fractions of the CBMs that detach from crystal surfaces or diffuse to other surfaces, the diffusivity along the hydrophobic surface, and the overall orientation of the CBM on both hydrophobic and hydrophilic faces. The simulations demonstrate that there is a thermodynamic driving force for the Cel7A CBM to bind preferentially to the hydrophobic surface of cellulose relative to hydrophilic surfaces. In addition, the simulations demonstrate that the CBM can diffuse from hydrophilic surfaces to the hydrophobic surface, whereas the reverse transition is not observed. Lastly, our simulations suggest that the flat faces of Family 1 CBMs are the preferred binding surfaces. These results enhance our understanding of how Family 1 CBMs interact with and recognize specific cellulose surfaces and provide insights into the initial events of cellulase adsorption and diffusion on cellulose.

  14. Periodic permanent magnet development for linear collider X-band klystrons

    SciTech Connect (OSTI)

    Sprehn, D.; Caryotakis, G.; Jongewaard, E.; Phillips, R. [Stanford Linear Accelerator Center, Stanford University, Stanford, California 94309 (United States)

    1999-05-07

    The Stanford Linear Accelerator Center (SLAC) klystron group is currently designing, fabricating and testing 11.424 GHz klystrons with peak output powers from 50 to 75 MW at 1 to 2 {mu}s rf pulsewidths as part of an effort to realize components necessary for the construction of the Next Linear Collider (NLC). In order to eliminate the projected operational-year energy bill for klystron solenoids, Periodic Permanent Magnet (PPM) focusing has been employed on our latest X-band klystron designs. A PPM beam tester has operated at the same repetition rate, voltage and average beam power required for a 75-MW NLC klystron. Prototype 50 and 75-MW PPM klystrons were built and tested during 1996 and 1997 which operate from 50 to 70 MW at efficiencies greater than 55%. Construction and testing of 75-MW research klystrons will continue while the design and reliability is perfected. This paper will discuss the design of these PPM klystrons and the results of testing to date along with future plans for the development of a low-cost Design for Manufacture (DFM) 75-MW klystron and invitation for industry participation.

  15. Periodic permanent magnet development for linear collider X-band klystrons

    SciTech Connect (OSTI)

    Sprehn, D.; Caryotakis, G.; Jongewaard, E.; Phillips, R. [Stanford Linear Accelerator Center, Stanford University, Stanford, California 94309 (United States)

    1999-05-01

    The Stanford Linear Accelerator Center (SLAC) klystron group is currently designing, fabricating and testing 11.424 GHz klystrons with peak output powers from 50 to 75 MW at 1 to 2 {mu}s rf pulsewidths as part of an effort to realize components necessary for the construction of the Next Linear Collider (NLC). In order to eliminate the projected operational-year energy bill for klystron solenoids, Periodic Permanent Magnet (PPM) focusing has been employed on our latest X-band klystron designs. A PPM beam tester has operated at the same repetition rate, voltage and average beam power required for a 75-MW NLC klystron. Prototype 50 and 75-MW PPM klystrons were built and tested during 1996 and 1997 which operate from 50 to 70 MW at efficiencies greater than 55{percent}. Construction and testing of 75-MW research klystrons will continue while the design and reliability is perfected. This paper will discuss the design of these PPM klystrons and the results of testing to date along with future plans for the development of a low-cost Design for Manufacture (DFM) 75-MW klystron and invitation for industry participation. {copyright} {ital 1999 American Institute of Physics.}

  16. Chemopreventive activity of compounds extracted from Casearia sylvestris (Salicaceae) Sw against DNA damage induced by particulate matter emitted by sugarcane burning near Araraquara, Brazil

    SciTech Connect (OSTI)

    Prieto, A.M.; Santos, A.G.; Csipak, A.R.; Caliri, C.M.; Silva, I.C.; Arbex, M.A.; Silva, F.S.; Marchi, M.R.R.

    2012-12-15

    Ethanolic extract of Casearia sylvestris is thought to be antimutagenic. In this study, we attempted to determine whether this extract and casearin X (a clerodane diterpene from C. sylvestris) are protective against the harmful effects of airborne pollutants from sugarcane burning. To that end, we used the Tradescantia micronucleus test in meiotic pollen cells of Tradescantia pallida, the micronucleus test in mouse bone marrow cells, and the comet assay in mouse blood cells. The mutagenic compound was total suspended particulate (TSP) from air. For the Tradescantia micronucleus test, T. pallida cuttings were treated with the extract at 0.13, 0.25, or 0.50 mg/ml. Subsequently, TSP was added at 0.3 mg/ml, and tetrads from the inflorescences were examined for micronuclei. For the micronucleus test in mouse bone marrow cells and the comet assay in mouse blood cells, Balb/c mice were treated for 15 days with the extract3.9, 7.5, or 15.0 mg/kg body weight (BW)or with casearin X0.3, 0.25, or 1.2 mg/kg BWafter which they received TSP (3.75 mg/kg BW). In T. pallida and mouse bone marrow cells, the extract was antimutagenic at all concentrations tested. In mouse blood cells, the extract was antigenotoxic at all concentrations, whereas casearin X was not antimutagenic but was antigenotoxic at all concentrations. We conclude that C. sylvestris ethanolic extract and casearin X protect DNA from damage induced by airborne pollutants from sugarcane burning. -- Highlights: ? We assessed DNA protection of C. sylvestris ethanolic extract. ? We assessed DNA protection of casearin X. ? We used Tradescantia pallida micronucleus test as screening. ? We used comet assay and micronucleus test in mice. ? The compounds protected DNA against sugar cane burning pollutants.

  17. SU-E-T-376: 3-D Commissioning for An Image-Guided Small Animal Micro- Irradiation Platform

    SciTech Connect (OSTI)

    Qian, X; Wuu, C; Admovics, J

    2014-06-01

    Purpose: A 3-D radiochromic plastic dosimeter has been used to cross-test the isocentricity of a high resolution image-guided small animal microirradiation platform. In this platform, the mouse stage rotating for cone beam CT imaging is perpendicular to the gantry rotation for sub-millimeter radiation delivery. A 3-D dosimeter can be used to verify both imaging and irradiation coordinates. Methods: A 3-D dosimeter and optical CT scanner were used in this study. In the platform, both mouse stage and gantry can rotate 360 with rotation axis perpendicular to each other. Isocentricity and coincidence of mouse stage and gantry rotations were evaluated using star patterns. A 3-D dosimeter was placed on mouse stage with center at platform isocenter approximately. For CBCT isocentricity, with gantry moved to 90, the mouse stage rotated horizontally while the x-ray was delivered to the dosimeter at certain angles. For irradiation isocentricity, the gantry rotated 360 to deliver beams to the dosimeter at certain angles for star patterns. The uncertainties and agreement of both CBCT and irradiation isocenters can be determined from the star patterns. Both procedures were repeated 3 times using 3 dosimeters to determine short-term reproducibility. Finally, dosimeters were scanned using optical CT scanner to obtain the results. Results: The gantry isocentricity is 0.9 0.1 mm and mouse stage rotation isocentricity is about 0.91 0.11 mm. Agreement between the measured isocenters of irradiation and imaging coordinates was determined. The short-term reproducibility test yielded 0.5 0.1 mm between the imaging isocenter and the irradiation isocenter, with a maximum displacement of 0.7 0.1 mm. Conclusion: The 3-D dosimeter can be very useful in precise verification of targeting for a small animal irradiation research. In addition, a single 3-D dosimeter can provide information in both geometric and dosimetric uncertainty, which is crucial for translational studies.

  18. Networked alpha and gamma spectral acquisition and analysis system

    SciTech Connect (OSTI)

    Wilcox, C.M.; Gross, J.M.

    1993-10-01

    This manual assumes a knowledge of (terminology used and a working familiarity with) the windowing system and mouse of the Sun computer workstation. See the appropriate Sun manuals for additional information. ALDO, the alpha detector control program, is used to control, monitor, and edit log information associated with the collection of alpha spectra. Actual data collection and control functions are performed by Mizar Real-Time computers for which ALDO acts as a friendly user command interface and status display. It is normally started as part of your login procedure, but may also be started from the ``NETSPEC Utilities`` submenu of the root menu. The root menu is obtained by pushing the right mouse button when the cursor is over the root window (background picture). To become a user of ALDO and the other programs in the NETSPEC system, contact the person who performs systems administration tasks for the Sun computers. Most user interaction with ALDO is by means of mouse manipulation of screen items such as buttons, checkboxes, and sliders. The action of pushing the left mouse button when the cursor is over an item is called selecting that item. The left mouse button is therefore called the select button. The right mouse button is the menu button because a limited number of options may be displayed when that button is pressed when the cursor is over an item with a triangle (inverted delta). In this document, names of selectable items are printed in bold when they are first mentioned or when emphasis is helpful. In general, items which do not apply to the current context are either disabled or made invisible in order to prevent selection.

  19. Introduction to High Performance Computers Richard Gerber NERSC User Services

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    What are the main parts of a computer? Merit Badge Requirements ... 4. Explain the following to your counselor: a. The five major parts of a computer. ... Boy Scouts of America Offer a Computers Merit Badge 5 What are the "5 major parts"? 6 Five Major Parts eHow.com Answers.com Fluther.com Yahoo! Wikipedia CPU CPU CPU CPU Motherboard RAM Monitor RAM RAM Power Supply Hard Drive Printer Storage Power Supply Removable Media Video Card Mouse Keyboard/ Mouse Video Card Secondary Storage

  20. Mechanism for Clastogenic Activity of Naphthalene

    SciTech Connect (OSTI)

    Buchholz, Bruce A.

    2015-09-29

    Naphthalene incubations form DNA adducts in vitro in a dose dependent manner in both mouse and rat tissues. Rodent tissue incubations with naphthalene indicate that naphthalene forms as many DNA adducts as Benzo(a)pyrene, a known DNA binding carcinogen. The mouse airway has the greatest number of DNA adducts, corresponding to the higher metabolic activation of naphthalene in this location. Both rat tissues, the rat olfactory (tumor target) and the airways (non-tumor target), have similar levels of NA-DNA adducts, indicating that short term measures of initial adduct formation do not directly correlate with sites of tumor formation in the NTP bioassays.

  1. Section CC

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    30 J-12-1 ATTACHMENT J-12 GOVERNMENT FURNISHED SERVICES AND INFORMATION TABLE J-12.1 GFS/I LIST FROM SECTION C (SOW) ID GFS/I GFS/I Due Contract Section GF0001 DOE will administer MOUs with other law enforcement agencies or other Federal agencies (e.g., U.S. Department of Defense [Yakima Training Center]). DOE will provide copies of MOUs and/or contracts to the MSC. As required C.2.1.1.1 GF0002 DOE will provide Federal Commissions for Hanford Patrol personnel. As required C.2.1.1.1 GF0003 DOE

  2. Sandia National Laboratories: Research: High Consequence, Automation, &

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Robotics: Advanced Manipulation Manipulation Robotics Homepage About Robotics Research & Development Advanced Controls Advanced Manipulation Mighty Mouse (M2) Sandia Hand Cybernetics High-Consequence Automation Perception and Decision Tools Unique Mobility Facilities Publications and Factsheets Robotics Image Gallery Robotics Videos Contact Robotics Research Advanced Manipulation Addressing robotics challenges The Sandia Hand has overcome issues that have prevented widespread adoption of

  3. Cell lines for the production of monoclonal antibodies to human glycophorin A

    DOE Patents [OSTI]

    Bigbee, William L.; Fong, Stella S. N.; Jensen, Ronald H.; Vanderlaan, Martin; Langlois, Richard G.

    1988-01-01

    Cloned mouse hybridoma cell lines have been established which continuously produce antibodies that differentiate between the M and N forms of human glycophorin A. These antibodies have potential application as human blood group reagents, as markers for terminally differentiated erythroid cells and as immunofluorescent labels of somatically variant human erythrocytes.

  4. Conformationally constrained farnesoid X receptor (FXR) agonists: Heteroaryl replacements of the naphthalene

    SciTech Connect (OSTI)

    Bass, Jonathan Y.; Caravella, Justin A.; Chen, Lihong; Creech, Katrina L.; Deaton, David N.; Madauss, Kevin P.; Marr, Harry B.; McFadyen, Robert B.; Miller, Aaron B.; Mills, Wendy Y.; Navas, III, Frank; Parks, Derek J.; Smalley, Jr., Terrence L.; Spearing, Paul K.; Todd, Dan; Williams, Shawn P.; Wisely, G. Bruce

    2014-08-13

    To improve on the drug properties of GSK8062 1b, a series of heteroaryl bicyclic naphthalene replacements were prepared. The quinoline 1c was an equipotent FXR agonist with improved drug developability parameters relative to 1b. In addition, analog 1c lowered body weight gain and serum glucose in a DIO mouse model of diabetes.

  5. Accommodations for Physical Disabilities | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Physical Disabilities Accommodations for Physical Disabilities ADA-107-2T.jpg The Department offers a variety of accommodations to individuals with mobility limitations. Available computer and telecommunications access products include: Sequential Keystroke Input Key Repeat Rate Control Keyboard Macros Alternative Keyboards Non-Keyboard Dependent Input Devices Word Prediction Packages Speech Recognition Robotic Devices Mouse Alternatives Keyguard Optical Character Recognition Speaker Phone

  6. Transcriptional Regulation of Apolipoprotein A5 Gene Expression by the Nuclear Receptor ROR alpha

    SciTech Connect (OSTI)

    Genoux, Annelise; Dehondt, Helene; Helleboid-Chapman, Audrey; Duhem, Christian; Hum, Dean W.; Martin, Genevieve; Pennacchio, Len; Staels, Bart; Fruchart-Najib, Jamila; Fruchart, Jean-Charles

    2004-10-01

    Apolipoprotein A5 has recently been identified as a crucial determinant of plasma triglyceride levels. Our results showed that RORa up-regulates human APOA5 but has no effect on mouse apoa5 promoter. These data suggest an additional important physiological role for RORa in the regulation of genes involved in plasma triglyceride homeostasis in human and probably in the development of atherosclerosis

  7. Secure video communications system

    DOE Patents [OSTI]

    Smith, Robert L.

    1991-01-01

    A secure video communications system having at least one command network formed by a combination of subsystems. The combination of subsystems to include a video subsystem, an audio subsystem, a communications subsystem, and a control subsystem. The video communications system to be window driven and mouse operated, and having the ability to allow for secure point-to-point real-time teleconferencing.

  8. T-547: Microsoft Windows Human Interface Device (HID) Vulnerability

    Broader source: Energy.gov [DOE]

    Microsoft Windows does not properly warn the user before enabling additional Human Interface Device (HID) functionality over USB, which allows user-assisted attackers to execute arbitrary programs via crafted USB data, as demonstrated by keyboard and mouse data sent by malware on a Smartphone that the user connected to the computer.

  9. 2001 - 11 | Jefferson Lab

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    1 Nov 2001 Sat, 2001-11-17 00:00 Jefferson Lab Gets New Chief: Leemann takes top post (Times-Dispatch) Sat, 2001-11-17 00:00 Leemann Officially Takes Over Peninsula's Jefferson Lab (The Virginian-Pilot) Mon, 2001-11-05 00:00 Lab is Working to Build a Better Mouse Camera

  10. MicroRNA-130b targets Fmr1 and regulates embryonic neural progenitor cell proliferation and differentiation

    SciTech Connect (OSTI)

    Gong, Xi; Zhang, Kunshan; Wang, Yanlu; Wang, Junbang; Cui, Yaru; Li, Siguang; Luo, Yuping

    2013-10-04

    Highlights: We found that the 3? UTR of the Fmr1 mRNA is a target of miR-130b. MiR-130b suppresses the expression of Fmr1 in mouse embryonic stem cell. MiR-130b alters the proliferation of mouse embryonic stem cell. MiR-130b alters fate specification of mouse embryonic stem cell. -- Abstract: Fragile X syndrome, one of the most common forms of inherited mental retardation, is caused by expansion of the CGG repeat in the 5?-untranslated region of the X-linked Fmr1 gene, which results in transcriptional silencing and loss of expression of its encoded protein FMRP. The loss of FMRP increases proliferation and alters fate specification in adult neural progenitor cells (aNPCs). However, little is known about Fmr1 mRNA regulation at the transcriptional and post-transcriptional levels. In the present study, we report that miR-130b regulated Fmr1 expression by directly targeting its 3?-untranslated region (3? UTR). Up-regulation of miR-130b in mouse embryonic neural progenitor cells (eNPCs) decreased Fmr1 expression, markedly increased eNPC proliferation and altered the differentiation tendency of eNPCs, suggesting that antagonizing miR-130b may be a new therapeutic entry point for treating Fragile X syndrome.

  11. Method and cell lines for the production of monoclonal antibodies to human glycophorin A

    DOE Patents [OSTI]

    Bigbee, W.L.; Fong, S.S.N.; Jensen, R.H.; Vanderlaan, M.

    Cloned mouse hybridoma cell lines have been established which continuously produce antibodies that differentiate between the M and N forms of human glycophorin A. These antibodies have potential application as human blood group reagents, as markers for terminally differentiated erythroid cells and as immunofluorescent labels of somatically variant human erythrocytes.

  12. Optimized design for PIGMI

    SciTech Connect (OSTI)

    Hansborough, L.; Hamm, R.; Stovall, J.; Swenson, D.

    1980-01-01

    PIGMI (Pion Generator for Medical Irradiations) is a compact linear proton accelerator design, optimized for pion production and cancer treatment use in a hospital environment. Technology developed during a four-year PIGMI Prototype experimental program allows the design of smaller, less expensive, and more reliable proton linacs. A new type of low-energy accelerating structure, the radio-frequency quadrupole (RFQ) has been tested; it produces an exceptionally good-quality beam and allows the use of a simple 30-kV injector. Average axial electric-field gradients of over 9 MV/m have been demonstrated in a drift-tube linac (DTL) structure. Experimental work is underway to test the disk-and-washer (DAW) structure, another new type of accelerating structure for use in the high-energy coupled-cavity linac (CCL). Sufficient experimental and developmental progress has been made to closely define an actual PIGMI. It will consist of a 30-kV injector, and RFQ linac to a proton energy of 2.5 MeV, a DTL linac to 125 MeV, and a CCL linac to the final energy of 650 MeV. The total length of the accelerator is 133 meters. The RFQ and DTL will be driven by a single 440-MHz klystron; the CCL will be driven by six 1320-MHz klystrons. The peak beam current is 28 mA. The beam pulse length is 60 ..mu..s at a 60-Hz repetition rate, resulting in a 100-..mu..A average beam current. The total cost of the accelerator is estimated to be approx. $10 million.

  13. SU-E-T-578: MCEBRT, A Monte Carlo Code for External Beam Treatment Plan Verifications

    SciTech Connect (OSTI)

    Chibani, O; Ma, C; Eldib, A

    2014-06-01

    Purpose: Present a new Monte Carlo code (MCEBRT) for patient-specific dose calculations in external beam radiotherapy. The code MLC model is benchmarked and real patient plans are re-calculated using MCEBRT and compared with commercial TPS. Methods: MCEBRT is based on the GEPTS system (Med. Phys. 29 (2002) 835846). Phase space data generated for Varian linac photon beams (6 15 MV) are used as source term. MCEBRT uses a realistic MLC model (tongue and groove, rounded ends). Patient CT and DICOM RT files are used to generate a 3D patient phantom and simulate the treatment configuration (gantry, collimator and couch angles; jaw positions; MLC sequences; MUs). MCEBRT dose distributions and DVHs are compared with those from TPS in absolute way (Gy). Results: Calculations based on the developed MLC model closely matches transmission measurements (pin-point ionization chamber at selected positions and film for lateral dose profile). See Fig.1. Dose calculations for two clinical cases (whole brain irradiation with opposed beams and lung case with eight fields) are carried out and outcomes are compared with the Eclipse AAA algorithm. Good agreement is observed for the brain case (Figs 2-3) except at the surface where MCEBRT dose can be higher by 20%. This is due to better modeling of electron contamination by MCEBRT. For the lung case an overall good agreement (91% gamma index passing rate with 3%/3mm DTA criterion) is observed (Fig.4) but dose in lung can be over-estimated by up to 10% by AAA (Fig.5). CTV and PTV DVHs from TPS and MCEBRT are nevertheless close (Fig.6). Conclusion: A new Monte Carlo code is developed for plan verification. Contrary to phantombased QA measurements, MCEBRT simulate the exact patient geometry and tissue composition. MCEBRT can be used as extra verification layer for plans where surface dose and tissue heterogeneity are an issue.

  14. Planar velocity and scalar concentration measurements in shock-accelerated,unstable fluid interfaces.

    SciTech Connect (OSTI)

    Goodenough, C.; Kumar, S.; Marr-Lyon, M.; Boyts, A.; Prestridge, K. P.; Rightley, P. M.; Tomkins, C. D.; Cannon, M. T.; Kamm, J. R.; Rider, William; Zoldi, C. A.; Orlicz, G.; Vorobieff, P. V.

    2004-01-01

    We report applications of several high-speed photographic techniques to diagnose fluid instability and the onset of turbulence in an ongoing experimental study of the evolution of shock-accelerated, heavy-gas cylinders. Results are at Reynolds numbers well above that associated with the turbulent and mixing transitions. Recent developments in diagnostics enable high-resolution, planar (2D) measurements of velocity fields (using particle image velocimetry, or PIV) and scalar concentration (using planar laser-induced fluorescence, or PLIF). The purpose of this work is to understand the basic science of complex, shock-driven flows and to provide high-quality data for code validation and development. The combination of these high-speed optical methods, PIV and PLIF, is setting a new standard in validating large codes for fluid simulations. The PIV velocity measurements provide quantitative evidence of transition to turbulence. In the PIV technique, a frame transfer camera with a 1 ms separation is used to image flows illuminated by two 10 ns laser pulses. Individual particles in a seeded flow are tracked from frame to frame to produce a velocity field. Dynamic PLIF measurements of the concentration field are high-resolution, quantitative dynamic data that reveal finely detailed structure at several instances after shock passage. These structures include those associated with the incipient secondary instability and late-time transition. Multiple instances of the flow are captured using a single frame Apogee camera and laser pulses with 140 {mu}s spacing. We describe tradeoffs of diagnostic instrumentation to provide PLIF images.

  15. Clinical Utility of the Modified Segmental Boost Technique for Treatment of the Pelvis and Inguinal Nodes

    SciTech Connect (OSTI)

    Moran, M.S.; Castrucci, W.A.; Ahmad, M.; Song, H.; Lund, M.W.; Mani, S.; Chamberlain, Daniel; Higgins, S.A.

    2010-03-15

    Purpose: Low-lying pelvic malignancies often require simultaneous radiation to pelvis and inguinal nodes. We previously reported improved homogeneity with the modified segmental boost technique (MSBT) compared to that with traditional methods, using phantom models. Here we report our institutional clinical experience with MSBT. Methods and Materials: MSBT patients from May 2001 to March 2007 were evaluated. Parameters analyzed included isocenter/multileaf collimation shifts, time per fraction (four fields), monitor units (MU)/fraction, femoral doses, maximal dose relative to body mass index, and inguinal node depth. In addition, a dosimetric comparison of the MSBT versus intensity modulated radiation therapy (IMRT) was conducted. Results: Of the 37 MSBT patients identified, 32 were evaluable. Port film adjustments were required in 6% of films. Median values for each analyzed parameter were as follows: MU/fraction, 298 (range, 226-348); delivery time, 4 minutes; inguinal depth, 4.5 cm; volume receiving 45 Gy (V45), 7%; V27.5, 87%; body mass index, 25 (range, 16.0-33.8). Inguinal dose was 100% in all cases; in-field inhomogeneity ranged from 111% to 118%. IMRT resulted in significantly decreased dose to normal tissue but required more time for treatment planning and a higher number of MUs (1,184 vs. 313 MU). Conclusions: In our clinical experience, the mono-isocentric MSBT provides a high degree of accuracy, improved homogeneity compared with traditional techniques, ease of simulation, treatment planning, treatment delivery, and acceptable femoral doses for pelvic/inguinal radiation fields requiring 45 to 50.4 Gy. In addition, the MSBT delivers a relatively uniform dose distribution throughout the treatment volume, despite varying body habitus. Clinical scenarios for the use of MSBT vs. intensity-modulated radiation therapy are discussed. To our knowledge, this is the first study reporting the utility of MSBT in the clinical setting.

  16. SU-E-T-378: Limits and Possibilities of a Simplistic Approach to Whole Breast Radiation Therapy Planning

    SciTech Connect (OSTI)

    Hipp, E; Osa, E; No, H; Jozsef, G; Rosman, M; Formenti, S

    2014-06-01

    Purpose: Challenges for radiation therapy in developing countries include unreliable infrastructure and high patient load. We propose a system to treat whole breast in the prone position without computed tomography and/or planning software. Methods: Six parameters are measured using calipers and levels with the patient prone in the treatment position. (1) The largest separation; (2) the angle that separation makes with the horizontal; (3) the separation 2 cm posterior to the nipple; (4) the vertical distance between these two separations; (5) the sup/inf length and (6) angle of the desired posterior field edge. The data in (5) (6) and (2) provide field length, collimator and gantry angles. Isocenter is set to the midpoint of (1), anterior jaw setting is 20cm (half-beam setup), and the dose is prescribed to a point 1.5 cm anterior to isocenter. MUs and wedge angles are calculated using an MU calculator and by requiring 100% dose at that point and 100-105% at the midpoint of (3). Measurements on 30 CT scans were taken to obtain the data 1-6. To test the resulting MU/wedge combinations, they were entered into Eclipse (Varian) and dose distributions were calculated. The MU/wedge combinations were recorded and tabulated. Results: Performing a dose volume histogram analysis, the contoured breast V95 was 90.5%, and the average V90 was 94.1%. The maximum dose never exceeded 114.5%, (average 108%). The lung V20 was <5% for 96.7%, and the heart V5 was <10% for 93.3% of our sample. Conclusion: A method to provide prone whole breast treatment without CT-planning was developed. The method provides reasonable coverage and normal tissue sparing. This approach is not recommended if imaging and planning capabilities are available; it was designed to specifically avoid the need for CT planning and should be reserved to clinics that need to avoid that step.

  17. Generalizable Class Solutions for Treatment Planning of Spinal Stereotactic Body Radiation Therapy

    SciTech Connect (OSTI)

    Weksberg, David C.; Palmer, Matthew B.; Vu, Khoi N.; Rebueno, Neal C.; Sharp, Hadley J.; Luo, Dershan; Yang, James N.; Shiu, Almon S.; Rhines, Laurence D.; McAleer, Mary Frances; Brown, Paul D.; Chang, Eric L.

    2012-11-01

    Purpose: Spinal stereotactic body radiation therapy (SBRT) continues to emerge as an effective therapeutic approach to spinal metastases; however, treatment planning and delivery remain resource intensive at many centers, which may hamper efficient implementation in clinical practice. We sought to develop a generalizable class solution approach for spinal SBRT treatment planning that would allow confidence that a given plan provides optimal target coverage, reduce integral dose, and maximize planning efficiency. Methods and Materials: We examined 91 patients treated with spinal SBRT at our institution. Treatment plans were categorized by lesion location, clinical target volume (CTV) configuration, and dose fractionation scheme, and then analyzed to determine the technically achievable dose gradient. A radial cord expansion was subtracted from the CTV to yield a planning CTV (pCTV) construct for plan evaluation. We reviewed the treatment plans with respect to target coverage, dose gradient, integral dose, conformality, and maximum cord dose to select the best plans and develop a set of class solutions. Results: The class solution technique generated plans that maintained target coverage and improved conformality (1.2-fold increase in the 95% van't Riet Conformation Number describing the conformality of a reference dose to the target) while reducing normal tissue integral dose (1.3-fold decrease in the volume receiving 4 Gy (V{sub 4Gy}) and machine output (19% monitor unit (MU) reduction). In trials of planning efficiency, the class solution technique reduced treatment planning time by 30% to 60% and MUs required by {approx}20%: an effect independent of prior planning experience. Conclusions: We have developed a set of class solutions for spinal SBRT that incorporate a pCTV metric for plan evaluation while yielding dosimetrically superior treatment plans with increased planning efficiency. Our technique thus allows for efficient, reproducible, and high-quality spinal SBRT treatment planning.

  18. SU-E-T-584: Commissioning of the MC2 Monte Carlo Dose Computation Engine

    SciTech Connect (OSTI)

    Titt, U; Mirkovic, D; Liu, A; Ciangaru, G; Mohan, R; Anand, A; Perles, L

    2014-06-01

    Purpose: An automated system, MC2, was developed to convert DICOM proton therapy treatment plans into a sequence MCNPX input files, and submit these to a computing cluster. MC2 converts the results into DICOM format, and any treatment planning system can import the data for comparison vs. conventional dose predictions. This work describes the data and the efforts made to validate the MC2 system against measured dose profiles and how the system was calibrated to predict the correct number of monitor units (MUs) to deliver the prescribed dose. Methods: A set of simulated lateral and longitudinal profiles was compared to data measured for commissioning purposes and during annual quality assurance efforts. Acceptance criteria were relative dose differences smaller than 3% and differences in range (in water) of less than 2 mm. For two out of three double scattering beam lines validation results were already published. Spot checks were performed to assure proper performance. For the small snout, all available measurements were used for validation vs. simulated data. To calibrate the dose per MU, the energy deposition per source proton at the center of the spread out Bragg peaks (SOBPs) was recorded for a set of SOBPs from each option. Subsequently these were then scaled to the results of dose per MU determination based on published methods. The simulations of the doses in the magnetically scanned beam line were also validated vs. measured longitudinal and lateral profiles. The source parameters were fine tuned to achieve maximum agreement with measured data. The dosimetric calibration was performed by scoring energy deposition per proton, and scaling the results to a standard dose measurement of a 10 x 10 x 10 cm3 volume irradiation using 100 MU. Results: All simulated data passed the acceptance criteria. Conclusion: MC2 is fully validated and ready for clinical application.

  19. The effect of diluent gases on ignition delay times in the shock tube and in the rapid compression machine

    SciTech Connect (OSTI)

    Wuermel, J.; Silke, E.J.; Curran, H.J.; O Conaire, M.S.; Simmie, J.M.

    2007-10-15

    The diluent gas used in the preparation of test fuel/oxygen mixtures is inert and does not take part in the chemical reaction. However, it does have an effect on the measured ignition delay time both in rapid compression machines and in shock tubes - argon decelerates ignition in the RCM, but accelerates it in the shock tube under some conditions. This opposite effect is due to the times scales involved in these experimental devices. Typical ignition delay times in the RCM are in the region of 1-200 ms, while those in the shock tube are much shorter (10-1000 {mu}s). Comparative RCM experiments and simulations for helium, argon, xenon, and nitrogen have shown extreme heat loss in the postcompression period, particularly for helium. Autoignition measurements of 2,3-dimethylpentane have highlighted a direct dependency of ignition delay time on the type of diluent used, where longer ignition delay time were recorded with argon. This increased ignition delay time is due to the extreme cooling of argon in the postcompression period. This observation was strengthened by comparative experiments with helium and argon, where the diluent effect was even stronger for helium, caused by its higher thermal conductivity. In the shock tube, the diluent effect is opposite to that in the RCM. For dilute mixtures of isooctane, calculations have predicted that mixtures with argon will ignite faster than those with nitrogen, based on the relative heat capacities of the two diluent gases. Overall, we conclude that the choice of diluent gases in experimental devices must be made with care, as ignition delay times can depend strongly on the type of diluent gas used. (author)

  20. SU-E-T-291: Sensitivity of a Simple 2D EPID in Vivo Dosimetry

    SciTech Connect (OSTI)

    Peca, S; Brown, D

    2014-06-01

    Purpose: As radiotherapy (RT) increases in complexity, so does motivation for in vivo dosimetry (IVD), which may detect errors such as: setup, beam shaping and dose delivered. We have recently developed an easy-toimplement method for two-dimensional IVD based on images taken with the electronic portal imaging device (EPID) in cine mode during treatment. The purpose of this work is to characterize its sensitivity to possible RT delivery errors. Methods: We introduced a series of modifications to a simple RT field (1010, 100MU, 300RR, 20cm homogeneous phantom) to simulate errors. These modifications included multi-leaf collimator (MLC) position, number of MUs, and collimator angle. We quantified the sensitivity to inhomogeneities by inserting variable amounts of solid lung and bone. Finally we delivered realistic fields to an anthropomorphic phantom to estimate sensitivity to gantry angle and setup errors. Results: Our EPIDIVD is sensitive to MLC positioning errors of 1mm and 3mm in the closed and open directions respectively, and to 3% MU variations. Sensitivity to collimator angle depends on field shape irregularity; in the case of a 10x10 field, we are sensitive to errors of 0.8. The sensitivity to inhomogeneities is limited by the nature of MV imaging: approximately 1% signal change is noted when switching 5cm of water to equal amounts of bone or lung. This suggests that the EPID-IVD is likely not sensitive to small setup or gantry angle errors, as confirmed by anthropomorphic tests. Conclusion: We have characterized a simple method of 2D dose reconstruction at isocenter depth inside the patient, which is sensitive to possible RT delivery errors. This method may be useful as a secondary safety check, to prevent large errors from being carried on to following fractions, and to record delivered dose. By using readily available hardware, it is easily implemented and may prove especially useful in centers with limited resources.

  1. A retrospective analysis for patient-specific quality assurance of volumetric-modulated arc therapy plans

    SciTech Connect (OSTI)

    Li, Guangjun; Wu, Kui; Peng, Guang; Zhang, Yingjie; Bai, Sen

    2014-01-01

    Volumetric-modulated arc therapy (VMAT) is now widely used clinically, as it is capable of delivering a highly conformal dose distribution in a short time interval. We retrospectively analyzed patient-specific quality assurance (QA) of VMAT and examined the relationships between the planning parameters and the QA results. A total of 118 clinical VMAT cases underwent pretreatment QA. All plans had 3-dimensional diode array measurements, and 69 also had ion chamber measurements. Dose distribution and isocenter point dose were evaluated by comparing the measurements and the treatment planning system (TPS) calculations. In addition, the relationship between QA results and several planning parameters, such as dose level, control points (CPs), monitor units (MUs), average field width, and average leaf travel, were also analyzed. For delivered dose distribution, a gamma analysis passing rate greater than 90% was obtained for all plans and greater than 95% for 100 of 118 plans with the 3%/3-mm criteria. The difference (mean ± standard deviation) between the point doses measured by the ion chamber and those calculated by TPS was 0.9% ± 2.0% for all plans. For all cancer sites, nasopharyngeal carcinoma and gastric cancer have the lowest and highest average passing rates, respectively. From multivariate linear regression analysis, the dose level (p = 0.001) and the average leaf travel (p < 0.001) showed negative correlations with the passing rate, and the average field width (p = 0.003) showed a positive correlation with the passing rate, all indicating a correlation between the passing rate and the plan complexity. No statistically significant correlation was found between MU or CP and the passing rate. Analysis of the results of dosimetric pretreatment measurements as a function of VMAT plan parameters can provide important information to guide the plan parameter setting and optimization in TPS.

  2. Study of TFTR D-T neutron spectra using natural diamond detectors

    SciTech Connect (OSTI)

    Roquemore, A.L.; Krasilnikov, A.V., Gorelenkov, N.N.

    1996-12-31

    Three Natural Diamond Detector (NDD) based spectrometers have been used for neutron spectra measurement during Deuterium-Tritium (D-T) experiments using high power Neutral Beam Injection (NBI) and Ton Cyclotron Resonance Heating (ICRH) on the Tokamak Fusion Test Reactor (TFTR) in 1996. A 2-3 % energy resolution coupled with the high radiation resistance of NDDs (5 x 10{sup 14}n/cm{sup 2}) makes them ideal for measuring the D-T neutron spectra in the high radiation environment of TFTR tritium experiments. The compact size of the NDD made it possible to insert one of the detectors into one of the center channels of the TFTR multichannel neutron collimator to provide a vertical view perpendicular to the vessel midplane, Two other detectors were placed inside shields in TFTR test cell and provide measurements of the neutrons having angles of emission of 110- 180{degrees} and 60-12-{degrees} with respect to the direction of the plasma current. By using a 0.25 {mu}s shaping time of the Ortec 673 spectroscopy amplifier we were able to accumulate useful spectrometry data at count rates up to 1.5 x 10{sup 3} counts/sec. To model the D- T neutron spectra measured by each of three NDD`s the Neutron Source post TRANSP (NST) code and semi-analytical model were developed. A set of D-T and D-D plasmas is analyzed for the dynamics of D-T neutron spectral broadening for each of three NDD cones of view. The application of three NDD based D-T neutron -spectrometer array demonstrated the anisotropy of the ion distribution function. and provided a mature of the dynamics of the effective ion temperatures for each detector view, and determined the tangential velocity of resonant tritons during ICRH.

  3. EFFECTS OF INTERMITTENT EMISSION: NOISE INVENTORY FOR THE SCINTILLATING PULSAR B0834+06

    SciTech Connect (OSTI)

    Gwinn, C. R.; Johnson, M. D.; Smirnova, T. V.; Stinebring, D. R. E-mail: michaeltdh@physics.ucsb.edu E-mail: dan.stinebring@oberlin.edu

    2011-05-20

    We compare signal and noise for observations of the scintillating pulsar B0834+06, using very long baseline interferometry and a single-dish spectrometer. Comparisons between instruments and with models suggest that amplitude variations of the pulsar strongly affect the amount and distribution of self-noise. We show that noise follows a quadratic polynomial with flux density, in spectral observations. Constant coefficients, indicative of background noise, agree well with expectation; whereas second-order coefficients, indicative of self-noise, are {approx}3 times values expected for a pulsar with constant on-pulse flux density. We show that variations in flux density during the 10 s integration accounts for the discrepancy. In the secondary spectrum, {approx}97% of spectral power lies within the pulsar's typical scintillation bandwidth and timescale; an extended scintillation arc contains {approx}3%. For a pulsar with constant on-pulse flux density, noise in the dynamic spectrum will appear as a uniformly distributed background in the secondary spectrum. We find that this uniform noise background contains 95% of noise in the dynamic spectrum for interferometric observations; but only 35% of noise in the dynamic spectrum for single-dish observations. Receiver and sky dominate noise for our interferometric observations, whereas self-noise dominates for single-dish. We suggest that intermittent emission by the pulsar, on timescales <300 {mu}s, concentrates self-noise near the origin in the secondary spectrum, by correlating noise over the dynamic spectrum. We suggest that intermittency sets fundamental limits on pulsar astrometry or timing. Accounting of noise may provide means for detection of intermittent sources, when effects of propagation are unknown or impractical to invert.

  4. PROSPECTS FOR PROBING THE SPACETIME OF Sgr A* WITH PULSARS

    SciTech Connect (OSTI)

    Liu, K.; Wex, N.; Kramer, M.; Cordes, J. M.; Lazio, T. J. W.

    2012-03-01

    The discovery of radio pulsars in compact orbits around Sgr A* would allow an unprecedented and detailed investigation of the spacetime of this supermassive black hole. This paper shows that pulsar timing, including that of a single pulsar, has the potential to provide novel tests of general relativity, in particular its cosmic censorship conjecture and no-hair theorem for rotating black holes. These experiments can be performed by timing observations with 100 {mu}s precision, achievable with the Square Kilometre Array for a normal pulsar at frequency above 15 GHz. Based on the standard pulsar timing technique, we develop a method that allows the determination of the mass, spin, and quadrupole moment of Sgr A*, and provides a consistent covariance analysis of the measurement errors. Furthermore, we test this method in detailed mock data simulations. It seems likely that only for orbital periods below {approx}0.3 yr is there the possibility of having negligible external perturbations. For such orbits, we expect a {approx}10{sup -3} test of the frame dragging and a {approx}10{sup -2} test of the no-hair theorem within five years, if Sgr A* is spinning rapidly. Our method is also capable of identifying perturbations caused by distributed mass around Sgr A*, thus providing high confidence in these gravity tests. Our analysis is not affected by uncertainties in our knowledge of the distance to the Galactic center, R{sub 0}. A combination of pulsar timing with the astrometric results of stellar orbits would greatly improve the measurement precision of R{sub 0}.

  5. A LIGHT CURVE ANALYSIS OF CLASSICAL NOVAE: FREE-FREE EMISSION VERSUS PHOTOSPHERIC EMISSION

    SciTech Connect (OSTI)

    Hachisu, Izumi; Kato, Mariko E-mail: mariko@educ.cc.keio.ac.jp

    2015-01-10

    We analyzed light curves of seven relatively slower novae, PW Vul, V705 Cas, GQ Mus, RR Pic, V5558 Sgr, HR Del, and V723 Cas, based on an optically thick wind theory of nova outbursts. For fast novae, free-free emission dominates the spectrum in optical bands rather than photospheric emission, and nova optical light curves follow the universal decline law. Faster novae blow stronger winds with larger mass-loss rates. Because the brightness of free-free emission depends directly on the wind mass-loss rate, faster novae show brighter optical maxima. In slower novae, however, we must take into account photospheric emission because of their lower wind mass-loss rates. We calculated three model light curves of free-free emission, photospheric emission, and their sum for various white dwarf (WD) masses with various chemical compositions of their envelopes and fitted reasonably with observational data of optical, near-IR (NIR), and UV bands. From light curve fittings of the seven novae, we estimated their absolute magnitudes, distances, and WD masses. In PW Vul and V705 Cas, free-free emission still dominates the spectrum in the optical and NIR bands. In the very slow novae, RR Pic, V5558 Sgr, HR Del, and V723 Cas, photospheric emission dominates the spectrum rather than free-free emission, which makes a deviation from the universal decline law. We have confirmed that the absolute brightnesses of our model light curves are consistent with the distance moduli of four classical novae with known distances (GK Per, V603 Aql, RR Pic, and DQ Her). We also discussed the reason why the very slow novae are about ∼1 mag brighter than the proposed maximum magnitude versus rate of decline relation.

  6. MO-G-BRE-01: A Real-Time Virtual Delivery System for Photon Radiotherapy Delivery Monitoring

    SciTech Connect (OSTI)

    Shi, F; Gu, X; Jiang, S; Jia, X; Graves, Y

    2014-06-15

    Purpose: Treatment delivery monitoring is important for radiotherapy, which enables catching dosimetric error at the earliest possible opportunity. This project develops a virtual delivery system to monitor the dose delivery process of photon radiotherapy in real-time using GPU-based Monte Carlo (MC) method. Methods: The simulation process consists of 3 parallel CPU threads. A thread T1 is responsible for communication with a linac, which acquires a set of linac status parameters, e.g. gantry angles, MLC configurations, and beam MUs every 20 ms. Since linac vendors currently do not offer interface to acquire data in real time, we mimic this process by fetching information from a linac dynalog file at the set frequency. Instantaneous beam fluence map (FM) is calculated. A FM buffer is also created in T1 and the instantaneous FM is accumulated to it. This process continues, until a ready signal is received from thread T2 on which an inhouse developed MC dose engine executes on GPU. At that moment, the accumulated FM is transferred to T2 for dose calculations, and the FM buffer in T1 is cleared. Once the calculation finishes, the resulting 3D dose distribution is directed to thread T3, which displays it in three orthogonal planes overlaid on the CT image for treatment monitoring. This process continues to monitor the 3D dose distribution in real-time. Results: An IMRT and a VMAT cases used in our patient-specific QA are studied. Maximum dose differences between our system and treatment planning system are 0.98% and 1.58% for the two cases, respectively. The average time per MC calculation is 0.1sec with <2% relative uncertainty. The update frequency of ∼10Hz is considered as real time. Conclusion: By embedding a GPU-based MC code in a novel data/work flow, it is possible to achieve real-time MC dose calculations to monitor delivery process.

  7. Measurements of charge state breeding efficiency at BNL test EBIS

    SciTech Connect (OSTI)

    Kondrashev, S.; Alessi, J.; Beebe, E.N.; Dickerson, C.; Ostroumov, P.N.; Pikin, A.; Savard, G.

    2011-04-02

    Charge breeding of singly charged ions is required to efficiently accelerate rare isotope ion beams for nuclear and astrophysics experiments, and to enhance the accuracy of low-energy Penning trap-assisted spectroscopy. An efficient charge breeder for the Californium Rare Isotope Breeder Upgrade (CARIBU) to the ANL Tandem Linear Accelerator System (ATLAS) facility is being developed using the BNL Test Electron Beam Ion Source (Test EBIS) as a prototype. Parameters of the CARIBU EBIS charge breeder are similar to those of the BNL Test EBIS except the electron beam current will be adjustable in the range from 1 to 2 {angstrom}. The electron beam current density in the CARIBU EBIS trap will be significantly higher than in existing operational charge state breeders based on the EBIS concept. The charge state breeding efficiency is expected to be about 25% for the isotope ions extracted from the CARIBU. For the success of our EBIS project, it is essential to demonstrate high breeding efficiency at the BNL Test EBIS tuned to the regime close to the parameters of the CARIBU EBIS at ANL. The breeding efficiency optimization and measurements have been successfully carried out using a Cs{sup +} surface ionization ion source for externally pulsed injection into the BNL Test EBIS. A Cs{sup +} ion beam with a total number of ions of 5 x 10{sup 8} and optimized pulse length of 70 {mu}s has been injected into the Test EBIS and charge-bred for 5.3 ms for two different electron beam currents 1 and 1.5 {angstrom}. In these experiments we have achieved 70% injection/extraction efficiency and breeding efficiency into the most abundant charge state 17%.

  8. Glucose cycling is markedly enhanced in pancreatic islets of obese hyperglycemic mice

    SciTech Connect (OSTI)

    Khan, A.; Chandramouli, V.; Ostenson, C.G.; Berggren, P.O.; Loew, H.L.; Landau, B.R.; Efendic, S. )

    1990-05-01

    Pancreatic islets from fed 7-month old lean and obese hyperglycemic mice (ob/ob) were incubated with 3H2O and 5.5 mM or 16.7 mM glucose. Incorporation of 3H into the medium glucose was taken as the measure of glucose-6-P hydrolysis to glucose. Glucose utilization was measured from the yield of 3H2O from (5-3H)glucose. Only 3-4% of the glucose phosphorylated was dephosphorylated by the lean mouse islets irrespective of the glucose concentration. In contrast, the ob/ob mouse islets at 5.5 mM glucose dephosphorylated 18% of the glucose phosphorylated and 30% at 16.7 mM. Thus, the islets of hyperglycemic mice demonstrate increased glucose cycling as compared to the islets of normoglycemic lean mice.

  9. Establishing a process of irradiating small animal brain using a CyberKnife and a microCT scanner

    SciTech Connect (OSTI)

    Kim, Haksoo; Welford, Scott; Fabien, Jeffrey; Zheng, Yiran; Yuan, Jake; Brindle, James; Yao, Min; Lo, Simon; Wessels, Barry; Machtay, Mitchell; Sohn, Jason W.; Sloan, Andrew

    2014-02-15

    Purpose: Establish and validate a process of accurately irradiating small animals using the CyberKnife G4 System (version 8.5) with treatment plans designed to irradiate a hemisphere of a mouse brain based on microCT scanner images. Methods: These experiments consisted of four parts: (1) building a mouse phantom for intensity modulated radiotherapy (IMRT) quality assurance (QA), (2) proving usability of a microCT for treatment planning, (3) fabricating a small animal positioning system for use with the CyberKnife's image guided radiotherapy (IGRT) system, and (4)in vivo verification of targeting accuracy. A set of solid water mouse phantoms was designed and fabricated, with radiochromic films (RCF) positioned in selected planes to measure delivered doses. After down-sampling for treatment planning compatibility, a CT image set of a phantom was imported into the CyberKnife treatment planning system—MultiPlan (ver. 3.5.2). A 0.5 cm diameter sphere was contoured within the phantom to represent a hemispherical section of a mouse brain. A nude mouse was scanned in an alpha cradle using a microCT scanner (cone-beam, 157 × 149 pixels slices, 0.2 mm longitudinal slice thickness). Based on the results of our positional accuracy study, a planning treatment volume (PTV) was created. A stereotactic body mold of the mouse was “printed” using a 3D printer laying UV curable acrylic plastic. Printer instructions were based on exported contours of the mouse's skin. Positional reproducibility in the mold was checked by measuring ten CT scans. To verify accurate dose delivery in vivo, six mice were irradiated in the mold with a 4 mm target contour and a 2 mm PTV margin to 3 Gy and sacrificed within 20 min to avoid DNA repair. The brain was sliced and stained for analysis. Results: For the IMRT QA using a set of phantoms, the planned dose (6 Gy to the calculation point) was compared to the delivered dose measured via film and analyzed using Gamma analysis (3% and 3 mm). A passing rate of 99% was measured in areas of above 40% of the prescription dose. The final inverse treatment plan was comprised of 43 beams ranging from 5 to 12.5 mm in diameter (2.5 mm size increments are available up to 15 mm in diameter collimation). Using the Xsight Spine Tracking module, the CyberKnife system could not reliably identify and track the tiny mouse spine; however, the CyberKnife system could identify and track the fiducial markers on the 3D mold.In vivo positional accuracy analysis using the 3D mold generated a mean error of 1.41 mm ± 0.73 mm when fiducial markers were used for position tracking. Analysis of the dissected brain confirmed the ability to target the correct brain volume. Conclusions: With the use of a stereotactic body mold with fiducial markers, microCT imaging, and resolution down-sampling, the CyberKnife system can successfully perform small-animal radiotherapy studies.

  10. Hybrid radiosity-SP{sub 3} equation based bioluminescence tomography reconstruction for turbid medium with low- and non-scattering regions

    SciTech Connect (OSTI)

    Chen, Xueli E-mail: jimleung@mail.xidian.edu.cn; Zhang, Qitan; Yang, Defu; Liang, Jimin E-mail: jimleung@mail.xidian.edu.cn

    2014-01-14

    To provide an ideal solution for a specific problem of gastric cancer detection in which low-scattering regions simultaneously existed with both the non- and high-scattering regions, a novel hybrid radiosity-SP{sub 3} equation based reconstruction algorithm for bioluminescence tomography was proposed in this paper. In the algorithm, the third-order simplified spherical harmonics approximation (SP{sub 3}) was combined with the radiosity equation to describe the bioluminescent light propagation in tissues, which provided acceptable accuracy for the turbid medium with both low- and non-scattering regions. The performance of the algorithm was evaluated with digital mouse based simulations and a gastric cancer-bearing mouse based in situ experiment. Primary results demonstrated the feasibility and superiority of the proposed algorithm for the turbid medium with low- and non-scattering regions.

  11. ChIP-seq Mapping of Distant-Acting Enhancers and Their In Vivo Activities

    SciTech Connect (OSTI)

    Visel, Axel; Pennacchio, Len A.

    2011-06-01

    The genomic location and function of most distant-acting transcriptional enhancers in the human genome remains unknown We performed ChIP-seq for various transcriptional coactivator proteins (such as p300) directly from different embryonic mouse tissues, identifying thousands of binding sitesTransgenic mouse experiments show that p300 and other co-activator peaks are highly predictive of genomic location AND tissue-specific activity patterns of distant-acting enhancersMost enhancers are active only in one or very few tissues Genomic location of tissue-specific p300 peaks correlates with tissue-specific expression of nearby genes Most binding sites are conserved, but the global degree of conservation varies between tissues

  12. Construction of highly extensive polymorphic DNA libraries by in-gel competitive reassociation procedure

    SciTech Connect (OSTI)

    Inoue, Shinichi; Kiyama, Ryoiti; Oishi, Michio

    1996-02-01

    Differential genomic DNA libraries between two mouse strains and from two human individuals were constructed by means of the in-gel competitive reassociation (IGCR) procedure, a procedure developed for cloning altered anonymous restriction fragments. The libraries were highly enriched fragments, approximately 60 and 40% for the mouse and human libraries, respectively, and, more importantly, maintained most of the original complexities of the RFLP fragments. Therefore, differential genomic DNA libraries constructed by the IGCR procedure, particularly for human genomic DNA, should offer highly extensive sources for polymorphic DNA sequences necessary for a variety of genome analyses, including studies on the origin and mechanism of biological diversity among the same species. 19 refs., 4 figs.

  13. Library Resources for Bac End Sequencing. Final Technical Report

    SciTech Connect (OSTI)

    Pieter J. de Jong

    2000-10-01

    Studies directed towards the specific aims outlined for this research award are summarized. The RPCI II Human Bac Library has been expanded by the addition of 6.9-fold genomic coverage. This segment has been generated from a MBOI partial digest of the same anonymous donor DNA used for the rest of the library. A new cloning vector, pTARBAC1, has been constructed and used in the construction of RPCI-II segment 5. This new cloning vector provides a new strategy in identifying targeted genomic regions and will greatly facilitate a large-scale analysis for positional cloning. A new maleCS7BC/6J mouse BAC library has been constructed. RPCI-23 contain 576 plates (approx 210,000 clones) and represents approximately 11-fold coverage of the mouse genome.

  14. Improved mutagen testing systems in mice

    SciTech Connect (OSTI)

    Roderick, T.H.

    1992-01-01

    Our laboratory was the first to induce and ascertain a mammalian chromosomal inversion; we did this by searching for a high frequency of first meiotic anaphase bridges in testes of males whose fathers received post-spermatogonial radiation or mutagenesis from chromosomal breaking chemical mutagens. One test in was examined in each mouse, and those showing a high frequency were then mated to determine if the high frequency were passed on as a dominant and whether linkage analysis suggested the presence of an inversion. A very high incidence (exceeding 20% bridges in first meiotic anaphase bridges) was found in about 1 in 150 males examined and this frequency was generally found to be passed on to the offspring an predicted. Later cytological banding techniques were developed elsewhere and we used them to show visually the inverted orders of the inverted chromosomal segments. Since that time we have induced inversions covering most of the mouse genome.

  15. Improved mutagen testing systems in mice. Final report

    SciTech Connect (OSTI)

    Roderick, T.H.

    1992-12-31

    Our laboratory was the first to induce and ascertain a mammalian chromosomal inversion; we did this by searching for a high frequency of first meiotic anaphase bridges in testes of males whose fathers received post-spermatogonial radiation or mutagenesis from chromosomal breaking chemical mutagens. One test in was examined in each mouse, and those showing a high frequency were then mated to determine if the high frequency were passed on as a dominant and whether linkage analysis suggested the presence of an inversion. A very high incidence (exceeding 20% bridges in first meiotic anaphase bridges) was found in about 1 in 150 males examined and this frequency was generally found to be passed on to the offspring an predicted. Later cytological banding techniques were developed elsewhere and we used them to show visually the inverted orders of the inverted chromosomal segments. Since that time we have induced inversions covering most of the mouse genome.

  16. Establishing a process of irradiating small animal brain using a CyberKnife and a microCT scanner

    SciTech Connect (OSTI)

    Kim, Haksoo; Welford, Scott; Fabien, Jeffrey; Zheng, Yiran; Yuan, Jake; Brindle, James; Yao, Min; Lo, Simon; Wessels, Barry; Machtay, Mitchell; Sohn, Jason W.; Sloan, Andrew

    2014-02-15

    Purpose: Establish and validate a process of accurately irradiating small animals using the CyberKnife G4 System (version 8.5) with treatment plans designed to irradiate a hemisphere of a mouse brain based on microCT scanner images. Methods: These experiments consisted of four parts: (1) building a mouse phantom for intensity modulated radiotherapy (IMRT) quality assurance (QA), (2) proving usability of a microCT for treatment planning, (3) fabricating a small animal positioning system for use with the CyberKnife's image guided radiotherapy (IGRT) system, and (4)in vivo verification of targeting accuracy. A set of solid water mouse phantoms was designed and fabricated, with radiochromic films (RCF) positioned in selected planes to measure delivered doses. After down-sampling for treatment planning compatibility, a CT image set of a phantom was imported into the CyberKnife treatment planning systemMultiPlan (ver. 3.5.2). A 0.5 cm diameter sphere was contoured within the phantom to represent a hemispherical section of a mouse brain. A nude mouse was scanned in an alpha cradle using a microCT scanner (cone-beam, 157 149 pixels slices, 0.2 mm longitudinal slice thickness). Based on the results of our positional accuracy study, a planning treatment volume (PTV) was created. A stereotactic body mold of the mouse was printed using a 3D printer laying UV curable acrylic plastic. Printer instructions were based on exported contours of the mouse's skin. Positional reproducibility in the mold was checked by measuring ten CT scans. To verify accurate dose delivery in vivo, six mice were irradiated in the mold with a 4 mm target contour and a 2 mm PTV margin to 3 Gy and sacrificed within 20 min to avoid DNA repair. The brain was sliced and stained for analysis. Results: For the IMRT QA using a set of phantoms, the planned dose (6 Gy to the calculation point) was compared to the delivered dose measured via film and analyzed using Gamma analysis (3% and 3 mm). A passing rate of 99% was measured in areas of above 40% of the prescription dose. The final inverse treatment plan was comprised of 43 beams ranging from 5 to 12.5 mm in diameter (2.5 mm size increments are available up to 15 mm in diameter collimation). Using the Xsight Spine Tracking module, the CyberKnife system could not reliably identify and track the tiny mouse spine; however, the CyberKnife system could identify and track the fiducial markers on the 3D mold.In vivo positional accuracy analysis using the 3D mold generated a mean error of 1.41 mm 0.73 mm when fiducial markers were used for position tracking. Analysis of the dissected brain confirmed the ability to target the correct brain volume. Conclusions: With the use of a stereotactic body mold with fiducial markers, microCT imaging, and resolution down-sampling, the CyberKnife system can successfully perform small-animal radiotherapy studies.

  17. Finite Element Results Visualization for Unstructured Grids

    Energy Science and Technology Software Center (OSTI)

    1996-07-15

    GRIZ is a general-purpose post-processing application supporting interactive visualization of finite element analysis results on unstructured grids. In addition to basic pseudocolor renderings of state variables over the mesh surface, GRIZ provides modern visualization techniques such as isocontours and isosurfaces, cutting planes, vector field display, and particle traces. GRIZ accepts both command-line and mouse-driven input, and is portable to virtually any UNIX platform which provides Motif and OpenGl libraries.

  18. Webcam for New NERSC Building

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Scientific and Technical Information Web-enabled Milestones Chart Over the last decade, numerous milestones have been achieved, including many "firsts" in government web search technology. Mouse over the year to view the related milestone. Web-enabled milestones pioneered by OSTI 1994 dot First DOE homepage 1997 dot First searchable full-text DOE results 1999 dot First parallel searching of databases and websites 2000 dot First searchable gateway to preprint servers around the

  19. Federal Requirements for the Web | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Federal Requirements for the Web Federal Requirements for the Web Federal laws and requirements govern the Office of Energy Efficiency and Renewable Energy (EERE) with regards to its websites and other digital media. Section 508 Section 508 of the Rehabilitation Act requires federal websites to make all content accessible to users with disabilities. Disabled users may use screen readers, or have hearing issues, colorblindness, or difficulties using traditional hardware like a mouse or keyboard.

  20. Memorandum of Understanding (MOU) | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    of Understanding (MOU) Memorandum of Understanding (MOU) The following document is a sample Memorandum of Understanding for DOE laboratories to use as guidance in drafting lab MOUs. PDF icon Sample_lab_MOU.pdf More Documents & Publications Memorandum of Understanding (MOU) Between the State of Nevada and the U.S. Department of Energy Memorandum of understanding between DOE and NGLIA Electric Power Research Institute Cooperation to Increase Energy Efficiency, March 6, 2008

  1. Supercomputing: Eye-Opening Possibilities in Imaging | Department of Energy

    Energy Savers [EERE]

    Supercomputing: Eye-Opening Possibilities in Imaging Supercomputing: Eye-Opening Possibilities in Imaging September 20, 2013 - 5:00pm Addthis This overlay of mass spectrometry images shows the spatial distribution of three different kind of lipids across a whole mouse cross-section. Lipids act as the structural components of cell membranes and are responsible for energy storage, among other things. | Photo courtesy of Wolfgang Reindl (Berkeley Lab). This overlay of mass spectrometry images shows

  2. NETLMemorandum of Understanding or Agreement

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Memorandum of Understanding or Agreement Memorandums of Understanding (MOUs) OR Memorandums of Agreement (MOAs) are written agreements between NETL and other entities that state of scope of work for a specific project or state the terms of a partnering relationship. Parties to these agreements may include other federal agencies, local, state, international, or other government entities; the private sector; and educational institutions. An MOU or MOA is not considered a binding contract. It

  3. 'Data Deluge' Pushes Mass Spec Imaging to New Heights

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    'Data Deluge' Pushes Mass Spec Imaging to New Heights 'Data Deluge' Pushes Mass Spec Imaging to New Heights MANTISSA Team Takes Novel Approach to Improve Experimental Data Analysis July 15, 2015 Contact: Kathy Kincade, +1 510 495 2124, kkincade@lbl.gov MANTISSA Ion-intensity visualization of the 20 most important ions in a mouse brain segment selected by the CX/CUR algorithm. Of the 20 ions, little redundancy is present, pointing to the effectiveness of the CX approach for information

  4. Radiolabelling and positron emission tomography of PT70, a time-dependent inhibitor of InhA, the Mycobacterium tuberculosis enoyl-ACP reductase

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wang, Hui; Liu, Li; Lu, Yang; Pan, Pan; Hooker, Jacob M.; Fowler, Joanna S.; Tonge, Peter J.

    2015-07-14

    PT70 is a diaryl ether inhibitor of InhA, the enoyl-ACP reductase in the Mycobacterium tuberculosis fatty acid biosynthesis pathway. It has a residence time of 24 min on the target, and also shows antibacterial activity in a mouse model of tuberculosis infection. Due to the interest in studying target tissue pharmacokinetics of PT70, we developed a method to radiolabel PT70 with carbon-11 and have studied its pharmacokinetics in mice and baboons using positron emission tomography.

  5. Finishing in the Future

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    8 (Completed) Areas emphasized at this conference included: Keynote speakers Sydney Brenner, 2002 Nobel Prize Winner for Medicine, Founder of the Molecular Sciences Institute, and Senior Distinguished Fellow of the Crick-Jacobs Center, Salk Institute Bruce Birren, Director of the Microbial Sequencing Center and Co-Director of the Genome Sequencing and Analysis Program, Broad Institute Deanna Church, Coordinator for the Mouse and Human Genome Resources and Trace Archive, National Center for

  6. Microsoft Word - S08568_CY2011 Annual Rpt

    Office of Legacy Management (LM)

    2 Spatial ecology data for the Site are available for several data types and are stored in the GIS on the servers in Grand Junction, Colorado. The types of ecological spatial data that are available include annual weed distribution data (for selected species), annual weed control locations, biocontrol release locations, vegetation and wildlife monitoring locations (transect endpoints and sample points), vegetation community classifications, Preble's mouse habitat, wetland locations,

  7. Targeted disruption of Ataxia-telangiectasia mutated gene in miniature pigs by somatic cell nuclear transfer

    SciTech Connect (OSTI)

    Kim, Young June; Ahn, Kwang Sung; Kim, Minjeong; Kim, Min Ju; Park, Sang-Min; Ryu, Junghyun; Ahn, Jin Seop; Heo, Soon Young; Kang, Jee Hyun; Choi, You Jung; Choi, Seong-Jun; Shim, Hosup

    2014-10-03

    Highlights: • ATM gene-targeted pigs were produced by somatic cell nuclear transfer. • A novel large animal model for ataxia telangiectasia was developed. • The new model may provide an alternative to the mouse model. - Abstract: Ataxia telangiectasia (A-T) is a recessive autosomal disorder associated with pleiotropic phenotypes, including progressive cerebellar degeneration, gonad atrophy, and growth retardation. Even though A-T is known to be caused by the mutations in the Ataxia telangiectasia mutated (ATM) gene, the correlation between abnormal cellular physiology caused by ATM mutations and the multiple symptoms of A-T disease has not been clearly determined. None of the existing ATM mouse models properly reflects the extent to which neurological degeneration occurs in human. In an attempt to provide a large animal model for A-T, we produced gene-targeted pigs with mutations in the ATM gene by somatic cell nuclear transfer. The disrupted allele in the ATM gene of cloned piglets was confirmed via PCR and Southern blot analysis. The ATM gene-targeted pigs generated in the present study may provide an alternative to the current mouse model for the study of mechanisms underlying A-T disorder and for the development of new therapies.

  8. Genetic maps of polymorphic DNA loci on rat chromosome 1

    SciTech Connect (OSTI)

    Ding, Yan-Ping; Remmers, E.F.; Longman, R.E.

    1996-09-01

    Genetic linkage maps of loci defined by polymorphic DNA markers on rat chromosome 1 were constructed by genotyping F2 progeny of F344/N x LEW/N, BN/SsN x LEW/N, and DA/Bkl x F344/Hsd inbred rat strains. In total, 43 markers were mapped, of which 3 were restriction fragment length polymorphisms and the others were simple sequence length polymorphisms. Nineteen of these markers were associated with genes. Six markers for five genes, {gamma}-aminobutyric acid receptor {beta}3 (Gabrb3), syntaxin 2 (Stx2), adrenergic receptor {beta}3 (Gabrb3), syntaxin 2 (Stx2), adrenergic receptor {beta}1 (Adrb1), carcinoembryonic antigen gene family member 1 (Cgm1), and lipogenic protein S14 (Lpgp), and 20 anonymous loci were not previously reported. Thirteen gene loci (Myl2, Aldoa, Tnt, Igf2, Prkcg, Cgm4, Calm3, Cgm3, Psbp1, Sa, Hbb, Ins1, and Tcp1) were previously mapped. Comparative mapping analysis indicated that the large portion of rat chromosome 1 is homologous to mouse chromosome 7, although the homologous to mouse chromosome 7, although the homologs of two rat genes are located on mouse chromosomes 17 and 19. Homologs of the rat chromosome 1 genes that we mapped are located on human chromosomes 6, 10, 11, 12, 15, 16, and 19. 38 refs., 1 fig., 3 tabs.

  9. Assessment of dedicated low-dose cardiac micro-CT reconstruction algorithms using the left ventricular volume of small rodents as a performance measure

    SciTech Connect (OSTI)

    Maier, Joscha; Sawall, Stefan; Kachelrie, Marc

    2014-05-15

    Purpose: Phase-correlated microcomputed tomography (micro-CT) imaging plays an important role in the assessment of mouse models of cardiovascular diseases and the determination of functional parameters as the left ventricular volume. As the current gold standard, the phase-correlated Feldkamp reconstruction (PCF), shows poor performance in case of low dose scans, more sophisticated reconstruction algorithms have been proposed to enable low-dose imaging. In this study, the authors focus on the McKinnon-Bates (MKB) algorithm, the low dose phase-correlated (LDPC) reconstruction, and the high-dimensional total variation minimization reconstruction (HDTV) and investigate their potential to accurately determine the left ventricular volume at different dose levels from 50 to 500 mGy. The results were verified in phantom studies of a five-dimensional (5D) mathematical mouse phantom. Methods: Micro-CT data of eight mice, each administered with an x-ray dose of 500 mGy, were acquired, retrospectively gated for cardiac and respiratory motion and reconstructed using PCF, MKB, LDPC, and HDTV. Dose levels down to 50 mGy were simulated by using only a fraction of the projections. Contrast-to-noise ratio (CNR) was evaluated as a measure of image quality. Left ventricular volume was determined using different segmentation algorithms (Otsu, level sets, region growing). Forward projections of the 5D mouse phantom were performed to simulate a micro-CT scan. The simulated data were processed the same way as the real mouse data sets. Results: Compared to the conventional PCF reconstruction, the MKB, LDPC, and HDTV algorithm yield images of increased quality in terms of CNR. While the MKB reconstruction only provides small improvements, a significant increase of the CNR is observed in LDPC and HDTV reconstructions. The phantom studies demonstrate that left ventricular volumes can be determined accurately at 500 mGy. For lower dose levels which were simulated for real mouse data sets, the HDTV algorithm shows the best performance. At 50 mGy, the deviation from the reference obtained at 500 mGy were less than 4%. Also the LDPC algorithm provides reasonable results with deviation less than 10% at 50 mGy while PCF and MKB reconstruction show larger deviations even at higher dose levels. Conclusions: LDPC and HDTV increase CNR and allow for quantitative evaluations even at dose levels as low as 50 mGy. The left ventricular volumes exemplarily illustrate that cardiac parameters can be accurately estimated at lowest dose levels if sophisticated algorithms are used. This allows to reduce dose by a factor of 10 compared to today's gold standard and opens new options for longitudinal studies of the heart.

  10. SU-E-T-361: Clinical Benefit of Automatic Beam Gating Mixed with Breath Hold in Radiation Therapy of Left Breast

    SciTech Connect (OSTI)

    Wu, J; Hill, G; Spiegel, J; Ye, J; Mehta, V

    2014-06-01

    Purpose: To investigate the clinical and dosimetric benefits of automatic gating of left breast mixed with breath-hold technique. Methods: Two Active Breathing Control systems, ABC2.0 and ABC3.0, were used during simulation and treatment delivery. The two systems are different such that ABC2.0 is a breath-hold system without beam control capability, while ABC3.0 has capability in both breath-hold and beam gating. At simulation, each patient was scanned twice: one with free breathing (FB) and one with breath hold through ABC. Treatment plan was generated on the CT with ABC. The same plan was also recalculated on the CT with FB. These two plans were compared to assess plan quality. For treatments with ABC2.0, beams with MU > 55 were manually split into multiple subfields. All subfields were identical and shared the total MU. For treatment with ABC3.0, beam splitting was unnecessary. Instead, treatment was delivered in gating mode mixed with breath-hold technique. Treatment delivery efficiency using the two systems was compared. Results: The prescribed dose was 50.4Gy at 1.8Gy/fraction. The maximum heart dose averaged over 10 patients was 46.02.5Gy and 24.512.2Gy for treatments with FB and with ABC respectively. The corresponding heart V10 was 13.23.6% and 1.01.6% respectively. The averaged MUs were 99.87.5 for LMT, 99.29.4 for LLT. For treatment with ABC2.0, normally the original beam was split into 2 subfields. The averaged total time to delivery all beams was 4.30.4min for treatments with ABC2.0 and 3.30.6min for treatments with ABC3.0 in gating mode. Conclusion: Treatment with ABC tremendously reduced heart dose. Compared to treatments with ABC2.0, gating with ABC3.0 reduced the total treatment time by 23%. Use of ABC3.0 improved the delivery efficiency, and eliminated the possibility of mistreatments. The latter may happen with ABC2.0 where beam is not terminated when breath signal falls outside of the treatment window.

  11. Dosimetric comparison of 3D conformal, IMRT, and V-MAT techniques for accelerated partial-breast irradiation (APBI)

    SciTech Connect (OSTI)

    Qiu, Jian-Jian; Chang, Zheng; Horton, Janet K.; Wu, Qing-Rong Jackie; Yoo, Sua; Yin, Fang-Fang

    2014-07-01

    The purpose is to dosimetrically compare the following 3 delivery techniques: 3-dimensional conformal radiation therapy (3D-CRT), intensity-modulated arc therapy (IMRT), and volumetric-modulated arc therapy (V-MAT) in the treatment of accelerated partial-breast irradiation (APBI). Overall, 16 patients with T1/2N0 breast cancer were treated with 3D-CRT (multiple, noncoplanar photon fields) on the RTOG 0413 partial-breast trial. These cases were subsequently replanned using static gantry IMRT and V-MAT technology to understand dosimetric differences among these 3 techniques. Several dosimetric parameters were used in plan quality evaluation, including dose conformity index (CI) and dose-volume histogram analysis of normal tissue coverage. Quality assurance studies including gamma analysis were performed to compare the measured and calculated dose distributions. The IMRT and V-MAT plans gave more conformal target dose distributions than the 3D-CRT plans (p < 0.05 in CI). The volume of ipsilateral breast receiving 5 and 10 Gy was significantly less using the V-MAT technique than with either 3D-CRT or IMRT (p < 0.05). The maximum lung dose and the ipsilateral lung volume receiving 10 (V{sub 10}) or 20 Gy (V{sub 20}) were significantly less with both V-MAT and IMRT (p < 0.05). The IMRT technique was superior to 3D-CRT and V-MAT of low dose distributions in ipsilateral lung (p < 0.05 in V{sub 5} and D{sub 5}). The total mean monitor units (MUs) for V-MAT (621.0 111.9) were 12.2% less than those for 3D-CRT (707.3 130.9) and 46.5% less than those for IMRT (1161.4 315.6) (p < 0.05). The average machine delivery time was 1.5 0.2 minutes for the V-MAT plans, 7.0 1.6 minutes for the 3D-CRT plans, and 11.5 1.9 minutes for the IMRT plans, demonstrating much less delivery time for V-MAT. Based on this preliminary study, V-MAT and IMRT techniques offer improved dose conformity as compared with 3D-CRT techniques without increasing dose to the ipsilateral lung. In terms of MU and delivery time, V-MAT is significantly more efficient for APBI than for conventional 3D-CRT and static-beam IMRT.

  12. Groundwaters of Florence (Italy): Trace element distribution and vulnerability of the aquifers

    SciTech Connect (OSTI)

    Bencini, A.; Ercolanelli, R.; Sbaragli, A.

    1993-11-01

    Geochemical and hydrogeological research has been carried out in Florence, to evaluate conductivity and main chemistry of groundwaters, the pattern of some possible pollutant chemical species (Fe, Mn, Cr, Cu, Pb, Zn, NO{sub 2}, NO{sub 3}), and the vulnerability of the aquifers. The plain is made up of Plio-Quaternary alluvial and lacustrine sediments for a maximum thickness of 600 m. Silts and clays, sometimes with lenses of sandy gravels, are dominant, while considerable deposits of sands, pebbles, and gravels occur along the course of the Arno river and its tributary streams, and represent the most important aquifer of the plain. Most waters show conductivity values around 1000-1200 {mu}S, and almost all of them have an alkaline-earth-bicarbonate chemical character. In western areas higher salt content of the groundwaters is evident. Heavy metal and NO{sub 2}, NO{sub 3} analyses point out that no important pollution phenomena affect the groundwaters; all mean values are below the maximum admissible concentration (MAC) for drinkable waters. Some anomalies of NO{sub 2}, NO{sub 3}, Fe, Mn, and Zn are present. The most plausible causes can be recognized in losses of the sewage system; use of nitrate compounds in agriculture; oxidation of well pipes. All the observations of Cr, Cu, and Pb are below the MAC; the median values of <3, 3.9, and 1.1 {mu}g/l, respectively, could be considered reference concentrations for groundwaters in calcareous lithotypes, under undisturbed natural conditions. Finally, a map of vulnerability shows that the areas near the Arno river are highly vulnerable, for the minimum thickness (or lacking) of sediments covering the aquifer. On the other hand, in the case of pollution, several factors not considered could significantly increase the self-purification capacity of the aquifer, such asdilution of groundwaters, bacteria oxidation of nitrogenous species, and sorption capacity of clay minerals and organic matter. 31 refs., 6 figs., 5 tabs.

  13. SU-E-T-224: Is Monte Carlo Dose Calculation Method Necessary for Cyberknife Brain Treatment Planning?

    SciTech Connect (OSTI)

    Wang, L; Fourkal, E; Hayes, S; Jin, L; Ma, C

    2014-06-01

    Purpose: To study the dosimetric difference resulted in using the pencil beam algorithm instead of Monte Carlo (MC) methods for tumors adjacent to the skull. Methods: We retrospectively calculated the dosimetric differences between RT and MC algorithms for brain tumors treated with CyberKnife located adjacent to the skull for 18 patients (total of 27 tumors). The median tumor sizes was 0.53-cc (range 0.018-cc to 26.2-cc). The absolute mean distance from the tumor to the skull was 2.11 mm (range - 17.0 mm to 9.2 mm). The dosimetric variables examined include the mean, maximum, and minimum doses to the target, the target coverage (TC) and conformality index. The MC calculation used the same MUs as the RT dose calculation without further normalization and 1% statistical uncertainty. The differences were analyzed by tumor size and distance from the skull. Results: The TC was generally reduced with the MC calculation (24 out of 27 cases). The average difference in TC between RT and MC was 3.3% (range 0.0% to 23.5%). When the TC was deemed unacceptable, the plans were re-normalized in order to increase the TC to 99%. This resulted in a 6.9% maximum change in the prescription isodose line. The maximum changes in the mean, maximum, and minimum doses were 5.4 %, 7.7%, and 8.4%, respectively, before re-normalization. When the TC was analyzed with regards to target size, it was found that the worst coverage occurred with the smaller targets (0.018-cc). When the TC was analyzed with regards to the distance to the skull, there was no correlation between proximity to the skull and TC between the RT and MC plans. Conclusions: For smaller targets (< 4.0-cc), MC should be used to re-evaluate the dose coverage after RT is used for the initial dose calculation in order to ensure target coverage.

  14. Probing for exoplanets hiding in dusty debris disks: Disk imaging, characterization, and exploration with HST/STIS multi-roll coronagraphy

    SciTech Connect (OSTI)

    Schneider, Glenn; Hinz, Phillip M.; Grady, Carol A.; Hines, Dean C.; Debes, John H.; Perrin, Marshall D.; Moro-Martin, Amaya; Stark, Christopher C.; Kuchner, Marc J.; Woodgate, Bruce E.; Weinberger, Alycia J.; Rodigas, Timothy J.; Wisniewski, John P.; Silverstone, Murray D.; Jang-Condell, Hannah; Henning, Thomas; Serabyn, Eugene; Tamura, Motohide

    2014-10-01

    Spatially resolved scattered-light images of circumstellar debris in exoplanetary systems constrain the physical properties and orbits of the dust particles in these systems. They also inform on co-orbiting (but unseen) planets, the systemic architectures, and forces perturbing the starlight-scattering circumstellar material. Using Hubble Space Telescope (HST)/Space Telescope Imaging Spectrograph (STIS) broadband optical coronagraphy, we have completed the observational phase of a program to study the spatial distribution of dust in a sample of 10 circumstellar debris systems and 1 'mature' protoplanetrary disk, all with HST pedigree, using point-spread-function-subtracted multi-roll coronagraphy. These observations probe stellocentric distances ?5 AU for the nearest systems, and simultaneously resolve disk substructures well beyond corresponding to the giant planet and Kuiper Belt regions within our own solar system. They also disclose diffuse very low-surface-brightness dust at larger stellocentric distances. Herein we present new results inclusive of fainter disks such as HD 92945 (F {sub disk}/F {sub star} = 5 10{sup 5}), confirming, and better revealing, the existence of a narrow inner debris ring within a larger diffuse dust disk. Other disks with ring-like substructures and significant asymmetries and complex morphologies include HD 181327, for which we posit a spray of ejecta from a recent massive collision in an exo-Kuiper Belt; HD 61005, suggested to be interacting with the local interstellar medium; and HD 15115 and HD 32297, also discussed in the context of putative environmental interactions. These disks and HD 15745 suggest that debris system evolution cannot be treated in isolation. For AU Mic's edge-on disk, we find out-of-plane surface brightness asymmetries at ?5 AU that may implicate the existence of one or more planetary perturbers. Time-resolved images of the MP Mus protoplanetary disk provide spatially resolved temporal variability in the disk illumination. These and other new images from our HST/STIS GO/12228 program enable direct inter-comparison of the architectures of these exoplanetary debris systems in the context of our own solar system.

  15. A new anti-tumor strategy based on in vivo tumstatin overexpression after plasmid electrotransfer in muscle

    SciTech Connect (OSTI)

    Thevenard, Jessica; Mir, Lluis M.; Dupont-Deshorgue, Aurélie; Monboisse, Jean-Claude; Brassart-Pasco, Sylvie

    2013-03-22

    Highlights: ► A new therapeutic strategy based on tumstatin in vivo overexpression is proposed. ► pVAX1©–tumstatin electrotransfer in muscle mediates protein expression in muscle. ► A substantial expression of tumstatin is detected in the serum of electrotransfected mice. ► Tumstatin overexpression decreases tumor growth and increases mouse survival. -- Abstract: The NC1 domains from the different α(IV) collagen chains were found to exert anti-tumorigenic and/or anti-angiogenic activities. A limitation to the therapeutic use of these matrikines is the large amount of purified recombinant proteins, in the milligram range in mice that should be administered daily throughout the experimental procedures. In the current study, we developed a new therapeutic approach based on tumstatin (NC1α3(IV)) overexpression in vivo in a mouse melanoma model. Gene electrotransfer of naked plasmid DNA (pDNA) is particularly attractive because of its simplicity, its lack of immune responsiveness and its safety. The pDNA electrotransfer in muscle mediates a substantial gene expression that lasts several months. A pVAX1© vector containing the tumstatin cDNA was injected into the legs of C57BL/6 mice and submitted to electrotranfer. Sera were collected at different times and tumstatin was quantified by ELISA. Tumstatin secretion reached a plateau at day 21 with an expression level of 12 μg/mL. For testing the effects of tumstatin expression on tumor growth in vivo, B16F1 melanoma cells were subcutaneously injected in mice 7 days after empty pVAX1© (Mock) or pVAX1©–tumstatin electrotransfer. Tumstatin expression triggered a large decrease in tumor growth and an increase in mouse survival. This new therapeutic approach seems promising to inhibit tumor progression in vivo.

  16. Analysis of lung tumor initiation and progression in transgenic mice for Cre-inducible overexpression of Cul4A gene

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; Hung, Ming -Szu; Hsieh, David; Au, Alfred; Jablons, David M.; You, Liang

    2015-06-08

    Background: Lung cancer is the leading cause of morbidity and death worldwide. Although the available lung cancer animal models have been informative and further propel our understanding of human lung cancer, they still do not fully recapitulate the complexities of human lung cancer. The pathogenesis of lung cancer remains highly elusive because of its aggressive biologic nature and considerable heterogeneity, compared to other cancers. The association of Cul4A amplification with aggressive tumor growth and poor prognosis has been suggested. Our previous study suggested that Cul4A is oncogenic in vitro, but its oncogenic role in vivo has not been studied. Methods:more » Viral delivery approaches have been used extensively to model cancer in mouse models. In our experiments, we used Cre-recombinase induced overexpression of the Cul4A gene in transgenic mice to study the role of Cul4A on lung tumor initiation and progression and have developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. Results: Here we show that the use of a recombinant adenovirus expressing Cre-recombinase (“AdenoCre”) to induce Cul4A overexpression in the lungs of mice allows controls of the timing and multiplicity of tumor initiation. Following our mouse models, we are able to study the potential role of Cul4A in the development and progression in pulmonary adenocarcinoma as well. Conclusion: Our findings indicate that Cul4A is oncogenic in vivo, and this mouse model is a tool in understanding the mechanisms of Cul4A in human cancers and for testing experimental therapies targeting Cul4A.« less

  17. A preliminary regional PBPK model of lung metabolism for improving species dependent descriptions of 1,3-butadiene and its metabolites

    SciTech Connect (OSTI)

    Campbell, Jerry; Van Landingham, Cynthia; Crowell, Susan; Gentry, Robinan; Kaden, Debra; Fiebelkorn, Stacy; Loccisano, Anne; Clewell, Harvey

    2015-06-12

    1,3-Butadiene (BD), a volatile organic chemical (VOC), is used in synthetic rubber production and other industrial processes. It is detectable at low levels in ambient air as well as in tobacco smoke and gasoline vapors. Inhalation exposures to high concentrations of BD have been associated with lung cancer in both humans and experimental animals, although differences in species sensitivity have been observed. Metabolically active lung cells such as Pulmonary Type I and Type II epithelial cells and club cells (Clara cells)1 are potential targets of BD metabolite-induced toxicity. Metabolic capacities of these cells, their regional densities, and distributions vary throughout the respiratory tract as well as between species and cell types. Here we present a physiologically based pharmacokinetic (PBPK) model for BD that includes a regional model of lung metabolism, based on a previous model for styrene, to provide species-dependent descriptions of BD metabolism in the mouse, rat, and human. Since there are no in vivo data on BD pharmacokinetics in the human, the rat and mouse models were parameterized to the extent possible on the basis of in vitro metabolic data. Where it was necessary to use in vivo data, extrapolation from rat to mouse was performed to evaluate the level of uncertainty in the human model. A kidney compartment and description of downstream metabolism were also included in the model to allow for eventual use of available urinary and blood biomarker data in animals and humans to calibrate the model for estimation of BD exposures and internal metabolite levels. Results from simulated inhalation exposures to BD indicate that incorporation of differential lung region metabolism is important in describing species differences in pulmonary response and that these differences may have implications for risk assessments of human exposures to BD.

  18. A preliminary regional PBPK model of lung metabolism for improving species dependent descriptions of 1,3-butadiene and its metabolites

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Campbell, Jerry; Van Landingham, Cynthia; Crowell, Susan; Gentry, Robinan; Kaden, Debra; Fiebelkorn, Stacy; Loccisano, Anne; Clewell, Harvey

    2015-06-12

    1,3-Butadiene (BD), a volatile organic chemical (VOC), is used in synthetic rubber production and other industrial processes. It is detectable at low levels in ambient air as well as in tobacco smoke and gasoline vapors. Inhalation exposures to high concentrations of BD have been associated with lung cancer in both humans and experimental animals, although differences in species sensitivity have been observed. Metabolically active lung cells such as Pulmonary Type I and Type II epithelial cells and club cells (Clara cells)1 are potential targets of BD metabolite-induced toxicity. Metabolic capacities of these cells, their regional densities, and distributions vary throughoutmore » the respiratory tract as well as between species and cell types. Here we present a physiologically based pharmacokinetic (PBPK) model for BD that includes a regional model of lung metabolism, based on a previous model for styrene, to provide species-dependent descriptions of BD metabolism in the mouse, rat, and human. Since there are no in vivo data on BD pharmacokinetics in the human, the rat and mouse models were parameterized to the extent possible on the basis of in vitro metabolic data. Where it was necessary to use in vivo data, extrapolation from rat to mouse was performed to evaluate the level of uncertainty in the human model. A kidney compartment and description of downstream metabolism were also included in the model to allow for eventual use of available urinary and blood biomarker data in animals and humans to calibrate the model for estimation of BD exposures and internal metabolite levels. Results from simulated inhalation exposures to BD indicate that incorporation of differential lung region metabolism is important in describing species differences in pulmonary response and that these differences may have implications for risk assessments of human exposures to BD.« less

  19. Mechanisms of G1 cell cycle arrest and apoptosis in myeloma cells induced by hybrid-compound histone deacetylase inhibitor

    SciTech Connect (OSTI)

    Fujii, Seiko; Division of Maxillofacial Surgery, Kyushu Dental University ; Okinaga, Toshinori; Ariyoshi, Wataru; Oral Biology Research Center, Kyushu Dental University ; Takahashi, Osamu; Iwanaga, Kenjiro; Nishino, Norikazu; Tominaga, Kazuhiro; Nishihara, Tatsuji; Oral Biology Research Center, Kyushu Dental University

    2013-05-10

    Highlights: •Novel histone deacetylase inhibitor Ky-2, remarkably inhibits myeloma cell growth. •Ky-2 demonstrates no cytotoxicity against normal lymphocytic cells. •Ky-2 induces cell cycle arrest through the cell cycle-associated proteins. •Ky-2 induces Bcl-2-inhibitable apoptosis through a caspase-dependent cascade. -- Abstract: Objectives: Histone deacetylase (HDAC) inhibitors are new therapeutic agents, used to treat various types of malignant cancers. In the present study, we investigated the effects of Ky-2, a hybrid-compound HDAC inhibitor, on the growth of mouse myeloma cells. Materials and methods: Myeloma cells, HS-72, P3U1, and mouse normal cells were used in this study. Effect of HDAC inhibitors on cell viability was determined by WST-assay and trypan blue assay. Cell cycle was analyzed using flow cytometer. The expression of cell cycle regulatory and the apoptosis associated proteins were examined by Western blot analysis. Hoechst’s staining was used to detect apoptotic cells. Results: Our findings showed that Ky-2 decreased the levels of HDACs, while it enhanced acetylation of histone H3. Myeloma cell proliferation was inhibited by Ky-2 treatment. Interestingly, Ky-2 had no cytotoxic effects on mouse normal cells. Ky-2 treatment induced G1-phase cell cycle arrest and accumulation of a sub-G1 phase population, while Western blotting analysis revealed that expressions of the cell cycle-associated proteins were up-regulated. Also, Ky-2 enhanced the cleavage of caspase-9 and -3 in myeloma cells, followed by DNA fragmentation. In addition, Ky-2 was not found to induce apoptosis in bcl-2 overexpressing myeloma cells. Conclusion: These findings suggest that Ky-2 induces apoptosis via a caspase-dependent cascade and Bcl-2-inhibitable mechanism in myeloma cells.

  20. Implausibility of the vibrational theory of olfaction

    SciTech Connect (OSTI)

    Block, Eric; Ertem, Mehmed Z.; Jang, Seogjoo; Matsunami, Hiroaki; Sekharan, Sivakumar; Dethier, Berenice; Gundala, Sivaji; Pan, Yi; Li, Shengju; Li, Zhen; Lodge, Stephene N.; Ozbil, Mehmet; Jiang, Huihong; Penalba, Sonia Flores; Batista, Victor S.; Zhuang, Hanyi

    2015-04-21

    The vibrational theory of olfaction assumes that electron transfer occurs across odorants at the active sites of odorant receptors (ORs), serving as a sensitive measure of odorant vibrational frequencies, ultimately leading to olfactory perception. A previous study reported that human subjects differentiated hydrogen/deuterium isotopomers (isomers with isotopic atoms) of the musk compound cyclopentadecanone as evidence supporting the theory. Here, we find no evidence for such differentiation at the molecular level. In fact, we find that the human musk-recognizing receptor, OR5AN1, identified using a heterologous OR expression system and robustly responding to cyclopentadecanone and muscone, fails to distinguish isotopomers of these compounds in vitro. Furthermore, the mouse (methylthio)methanethiol (MTMT)-recognizing receptor, MOR244-3, and other selected human and mouse ORs, responded similarly to normal, deuterated, and ¹³C isotopomers of their respective ligands, paralleling our results with the musk receptor OR5AN1. These findings suggest that the proposed vibration theory does not apply to the human musk receptor OR5AN1, mouse thiol receptor MOR244-3, or other ORs examined. Also, contrary to the vibration theory predictions, muscone-d₃₀ lacks the 1,380-1,550 cm⁻¹ IR bands claimed to be essential for musk odor. Furthermore, our theoretical analysis shows that the proposed electron transfer mechanism of the vibrational frequencies of odorants could be easily suppressed by quantum effects of non-odorant molecular vibrational modes. As a result, these and other concerns about electron transfer at ORs, together with our extensive experimental data, argue against the plausibility of the vibration theory.

  1. Differential metabolism of 4-hydroxynonenal in liver, lung and brain of mice and rats

    SciTech Connect (OSTI)

    Zheng, Ruijin; Dragomir, Ana-Cristina; Mishin, Vladimir; Richardson, Jason R.; Heck, Diane E.; Laskin, Debra L.; Laskin, Jeffrey D.

    2014-08-15

    The lipid peroxidation end-product 4-hydroxynonenal (4-HNE) is generated in tissues during oxidative stress. As a reactive aldehyde, it forms Michael adducts with nucleophiles, a process that disrupts cellular functioning. Liver, lung and brain are highly sensitive to xenobiotic-induced oxidative stress and readily generate 4-HNE. In the present studies, we compared 4-HNE metabolism in these tissues, a process that protects against tissue injury. 4-HNE was degraded slowly in total homogenates and S9 fractions of mouse liver, lung and brain. In liver, but not lung or brain, NAD(P)+ and NAD(P)H markedly stimulated 4-HNE metabolism. Similar results were observed in rat S9 fractions from these tissues. In liver, lung and brain S9 fractions, 4-HNE formed protein adducts. When NADH was used to stimulate 4-HNE metabolism, the formation of protein adducts was suppressed in liver, but not lung or brain. In both mouse and rat tissues, 4-HNE was also metabolized by glutathione S-transferases. The greatest activity was noted in livers of mice and in lungs of rats; relatively low glutathione S-transferase activity was detected in brain. In mouse hepatocytes, 4-HNE was rapidly taken up and metabolized. Simultaneously, 4-HNE-protein adducts were formed, suggesting that 4-HNE metabolism in intact cells does not prevent protein modifications. These data demonstrate that, in contrast to liver, lung and brain have a limited capacity to metabolize 4-HNE. The persistence of 4-HNE in these tissues may increase the likelihood of tissue injury during oxidative stress. - Highlights: • Lipid peroxidation generates 4-hydroxynonenal, a highly reactive aldehyde. • Rodent liver, but not lung or brain, is efficient in degrading 4-hydroxynonenal. • 4-hydroxynonenal persists in tissues with low metabolism, causing tissue damage.

  2. The PDZ-binding motif of Yes-associated protein is required for its co-activation of TEAD-mediated CTGF transcription and oncogenic cell transforming activity

    SciTech Connect (OSTI)

    Shimomura, Tadanori; Miyamura, Norio; Hata, Shoji; Miura, Ryota; Hirayama, Jun Nishina, Hiroshi

    2014-01-17

    Highlights: Loss of the PDZ-binding motif inhibits constitutively active YAP (5SA)-induced oncogenic cell transformation. The PDZ-binding motif of YAP promotes its nuclear localization in cultured cells and mouse liver. Loss of the PDZ-binding motif inhibits YAP (5SA)-induced CTGF transcription in cultured cells and mouse liver. -- Abstract: YAP is a transcriptional co-activator that acts downstream of the Hippo signaling pathway and regulates multiple cellular processes, including proliferation. Hippo pathway-dependent phosphorylation of YAP negatively regulates its function. Conversely, attenuation of Hippo-mediated phosphorylation of YAP increases its ability to stimulate proliferation and eventually induces oncogenic transformation. The C-terminus of YAP contains a highly conserved PDZ-binding motif that regulates YAPs functions in multiple ways. However, to date, the importance of the PDZ-binding motif to the oncogenic cell transforming activity of YAP has not been determined. In this study, we disrupted the PDZ-binding motif in the YAP (5SA) protein, in which the sites normally targeted by Hippo pathway-dependent phosphorylation are mutated. We found that loss of the PDZ-binding motif significantly inhibited the oncogenic transformation of cultured cells induced by YAP (5SA). In addition, the increased nuclear localization of YAP (5SA) and its enhanced activation of TEAD-dependent transcription of the cell proliferation gene CTGF were strongly reduced when the PDZ-binding motif was deleted. Similarly, in mouse liver, deletion of the PDZ-binding motif suppressed nuclear localization of YAP (5SA) and YAP (5SA)-induced CTGF expression. Taken together, our results indicate that the PDZ-binding motif of YAP is critical for YAP-mediated oncogenesis, and that this effect is mediated by YAPs co-activation of TEAD-mediated CTGF transcription.

  3. Structure-based discovery of NANOG variant with enhanced properties to promote self-renewal and reprogramming of pluripotent stem cells

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Hayashi, Yohei; Caboni, Laura; Das, Debanu; Yumoto, Fumiaki; Clayton, Thomas; Deller, Marc C.; Nguyen, Phuong; Farr, Carol L.; Chiu, Hsiu -Ju; Miller, Mitchell D.; et al

    2015-03-30

    NANOG (from Irish mythology Tír na nÓg) transcription factor plays a central role in maintaining pluripotency, cooperating with OCT4 (also known as POU5F1 or OCT3/4), SOX2, and other pluripotency factors. Although the physiological roles of the NANOG protein have been extensively explored, biochemical and biophysical properties in relation to its structural analysis are poorly understood. Here we determined the crystal structure of the human NANOG homeodomain (hNANOG HD) bound to an OCT4 promoter DNA, which revealed amino acid residues involved in DNA recognition that are likely to be functionally important. We generated a series of hNANOG HD alanine substitution mutantsmore » based on the protein–DNA interaction and evolutionary conservation and determined their biological activities. Some mutant proteins were less stable, resulting in loss or decreased affinity for DNA binding. Overexpression of the orthologous mouse NANOG (mNANOG) mutants failed to maintain self-renewal of mouse embryonic stem cells without leukemia inhibitory factor. These results suggest that these residues are critical for NANOG transcriptional activity. Interestingly, one mutant, hNANOG L122A, conversely enhanced protein stability and DNA-binding affinity. The mNANOG L122A, when overexpressed in mouse embryonic stem cells, maintained their expression of self-renewal markers even when retinoic acid was added to forcibly drive differentiation. When overexpressed in epiblast stem cells or human induced pluripotent stem cells, the L122A mutants enhanced reprogramming into ground-state pluripotency. These findings indicate that structural and biophysical information on key transcriptional factors provides insights into the manipulation of stem cell behaviors and a framework for rational protein engineering.« less

  4. Structure-based discovery of NANOG variant with enhanced properties to promote self-renewal and reprogramming of pluripotent stem cells

    SciTech Connect (OSTI)

    Hayashi, Yohei; Caboni, Laura; Das, Debanu; Yumoto, Fumiaki; Clayton, Thomas; Deller, Marc C.; Nguyen, Phuong; Farr, Carol L.; Chiu, Hsiu -Ju; Miller, Mitchell D.; Elsliger, Marc -André; Deacon, Ashley M.; Godzik, Adam; Lesley, Scott A.; Tomoda, Kiichiro; Conklin, Bruce R.; Wilson, Ian A.; Yamanaka, Shinya; Fletterick, Robert J.

    2015-03-30

    NANOG (from Irish mythology Tír na nÓg) transcription factor plays a central role in maintaining pluripotency, cooperating with OCT4 (also known as POU5F1 or OCT3/4), SOX2, and other pluripotency factors. Although the physiological roles of the NANOG protein have been extensively explored, biochemical and biophysical properties in relation to its structural analysis are poorly understood. Here we determined the crystal structure of the human NANOG homeodomain (hNANOG HD) bound to an OCT4 promoter DNA, which revealed amino acid residues involved in DNA recognition that are likely to be functionally important. We generated a series of hNANOG HD alanine substitution mutants based on the protein–DNA interaction and evolutionary conservation and determined their biological activities. Some mutant proteins were less stable, resulting in loss or decreased affinity for DNA binding. Overexpression of the orthologous mouse NANOG (mNANOG) mutants failed to maintain self-renewal of mouse embryonic stem cells without leukemia inhibitory factor. These results suggest that these residues are critical for NANOG transcriptional activity. Interestingly, one mutant, hNANOG L122A, conversely enhanced protein stability and DNA-binding affinity. The mNANOG L122A, when overexpressed in mouse embryonic stem cells, maintained their expression of self-renewal markers even when retinoic acid was added to forcibly drive differentiation. When overexpressed in epiblast stem cells or human induced pluripotent stem cells, the L122A mutants enhanced reprogramming into ground-state pluripotency. These findings indicate that structural and biophysical information on key transcriptional factors provides insights into the manipulation of stem cell behaviors and a framework for rational protein engineering.

  5. Analysis of lung tumor initiation and progression in transgenic mice for Cre-inducible overexpression of Cul4A gene

    SciTech Connect (OSTI)

    Wang, Yang; Xu, Zhidong; Mao, Jian -Hua; Hung, Ming -Szu; Hsieh, David; Au, Alfred; Jablons, David M.; You, Liang

    2015-06-08

    Background: Lung cancer is the leading cause of morbidity and death worldwide. Although the available lung cancer animal models have been informative and further propel our understanding of human lung cancer, they still do not fully recapitulate the complexities of human lung cancer. The pathogenesis of lung cancer remains highly elusive because of its aggressive biologic nature and considerable heterogeneity, compared to other cancers. The association of Cul4A amplification with aggressive tumor growth and poor prognosis has been suggested. Our previous study suggested that Cul4A is oncogenic in vitro, but its oncogenic role in vivo has not been studied. Methods: Viral delivery approaches have been used extensively to model cancer in mouse models. In our experiments, we used Cre-recombinase induced overexpression of the Cul4A gene in transgenic mice to study the role of Cul4A on lung tumor initiation and progression and have developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. Results: Here we show that the use of a recombinant adenovirus expressing Cre-recombinase (AdenoCre) to induce Cul4A overexpression in the lungs of mice allows controls of the timing and multiplicity of tumor initiation. Following our mouse models, we are able to study the potential role of Cul4A in the development and progression in pulmonary adenocarcinoma as well. Conclusion: Our findings indicate that Cul4A is oncogenic in vivo, and this mouse model is a tool in understanding the mechanisms of Cul4A in human cancers and for testing experimental therapies targeting Cul4A.

  6. OSHA's approach to risk assessment for setting a revised occupational exposure standard for 1,3-butadiene

    SciTech Connect (OSTI)

    Grossman, E.A.; Martonik, J. )

    1990-06-01

    In its 1980 benzene decision (Industrial Union Department, ALF-CIO v. American Petroleum Institute, 448 U.S. 607 (1980)), the Supreme Court ruled that before he can promulgate any permanent health or safety standard, the Secretary (of Labor) is required to make a threshold finding that a place of employment is unsafe--in the sense that significant risks are present and can be lessened by a change in practices (448 U.S. at 642). The Occupational Safety and Health Administration (OSHA) has interpreted this to mean that whenever possible, it must quantify the risk associated with occupational exposure to a toxic substance at the current permissible exposure limit (PEL). If OSHA determines that there is significant risk to workers' health at its current standard, then it must quantify the risk associated with a variety of alternative standards to determine at what level, if any, occupational exposure to a substance no longer poses a significant risk. For rulemaking on occupational exposure to 1,3-butadiene, there are two studies that are suitable for quantitative risk assessment. One is a mouse inhalation bioassay conducted by the National Toxicology Program (NTP), and the other is a rat inhalation bioassay conducted by Hazelton Laboratories Europe. Of the four risk assessments that have been submitted to OSHA, all four have used the mouse and/or rat data with a variety of models to quantify the risk associated with occupational exposure to 1,3-butadiene. In addition, OSHA has performed its own risk assessment using the female mouse and female rat data and the one-hit and multistage models.

  7. Implausibility of the vibrational theory of olfaction

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Block, Eric; Ertem, Mehmed Z.; Jang, Seogjoo; Matsunami, Hiroaki; Sekharan, Sivakumar; Dethier, Berenice; Gundala, Sivaji; Pan, Yi; Li, Shengju; Li, Zhen; et al

    2015-04-21

    The vibrational theory of olfaction assumes that electron transfer occurs across odorants at the active sites of odorant receptors (ORs), serving as a sensitive measure of odorant vibrational frequencies, ultimately leading to olfactory perception. A previous study reported that human subjects differentiated hydrogen/deuterium isotopomers (isomers with isotopic atoms) of the musk compound cyclopentadecanone as evidence supporting the theory. Here, we find no evidence for such differentiation at the molecular level. In fact, we find that the human musk-recognizing receptor, OR5AN1, identified using a heterologous OR expression system and robustly responding to cyclopentadecanone and muscone, fails to distinguish isotopomers of thesemore » compounds in vitro. Furthermore, the mouse (methylthio)methanethiol (MTMT)-recognizing receptor, MOR244-3, and other selected human and mouse ORs, responded similarly to normal, deuterated, and ¹³C isotopomers of their respective ligands, paralleling our results with the musk receptor OR5AN1. These findings suggest that the proposed vibration theory does not apply to the human musk receptor OR5AN1, mouse thiol receptor MOR244-3, or other ORs examined. Also, contrary to the vibration theory predictions, muscone-d₃₀ lacks the 1,380-1,550 cm⁻¹ IR bands claimed to be essential for musk odor. Furthermore, our theoretical analysis shows that the proposed electron transfer mechanism of the vibrational frequencies of odorants could be easily suppressed by quantum effects of non-odorant molecular vibrational modes. As a result, these and other concerns about electron transfer at ORs, together with our extensive experimental data, argue against the plausibility of the vibration theory.« less

  8. ECRbase: Database of Evolutionary Conserved Regions, Promoters, and Transcription Factor Binding Sites in Vertebrate Genomes

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Loots, Gabriela G. [LLNL; Ovcharenko, I. [LLNL

    Evolutionary conservation of DNA sequences provides a tool for the identification of functional elements in genomes. This database of evolutionary conserved regions (ECRs) in vertebrate genomes features a database of syntenic blocks that recapitulate the evolution of rearrangements in vertebrates and a comprehensive collection of promoters in all vertebrate genomes generated using multiple sources of gene annotation. The database also contains a collection of annotated transcription factor binding sites (TFBSs) in evolutionary conserved and promoter elements. ECRbase currently includes human, rhesus macaque, dog, opossum, rat, mouse, chicken, frog, zebrafish, and fugu genomes. (taken from paper in Journal: Bioinformatics, November 7, 2006, pp. 122-124

  9. PPPL Scientists bring mysterious process down to earth | Princeton Plasma

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Physics Lab Bottling Magnetic Reconnection: PPPL Scientists bring mysterious process down to earth By John Greenwald September 30, 2011 Tweet Widget Google Plus One Share on Facebook With the click of a computer mouse, a scientist at the U.S. Department of Energy's Princeton Plasma Physics Laboratory (PPPL) sends 10,000 volts of electricity into a chamber filled with hydrogen gas. The charge heats the gas to 100,000 degrees Centigrade. In an instant -- one-thousandth of a second, to be

  10. Technology Transfer Reporting Form

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    This form is to be completed by the TTO for individual inquiry/case activity during the quarter as required by the Technology Transfer Commercialization Act of 2000. Mouse over definitions and descriptions appear over text/check boxes where appropriate. After completing this form, click on the submit button. *If you have no TTO activity for the quarter, please fill in your name, FY and quarter, lab or facility and check the box "No Quarterly Activity". Initial Ombuds Contact:

  11. Creating a Tiny Human Body on a Chip

    ScienceCinema (OSTI)

    Hunsberger, Maren; Soscia, Dave; Moya, Monica

    2016-03-16

    LLNL science communicator Maren Hunsberger takes us "Inside the Lab" to learn about the iChip (In-vitro Chip-based Human Investigational Platform) project at Lawrence Livermore National Laboratory. "One application of the iChip system would be to develop new pharmaceutical drugs," explains Dave Soscia, LLNL postdoc. "When you test in a mouse for example, it's not as close to the human system as you can get. If we can take human cells and put them on devices and actually mimic the structure and function of the organ systems in the human, we can actually replace animal testing and even make a better system for testing pharmaceutical drugs."

  12. Field Deployable Tritium Assay System Host Graphical User Interface Software

    Energy Science and Technology Software Center (OSTI)

    1998-05-12

    The FDTASHOST software is a Graphical User Interface for the Field Deployable Tritium Assay System (FDTAS - Invention Disclosure SRS-96-09-091 has been submitted). The program runs on the Host computer which is located in the Laboratory and connected to the FDTAS remote field system via a modem over a phone line. The operator receives status information and messages from the Remote system. The operator can enter in commands to be executed by the remote systemmore » using the mouse and a pull down menu.« less

  13. GenoGraphics for OpenWindows trademark

    SciTech Connect (OSTI)

    Hagstrom, R.; Overbeek, R.; Price, M.; Zawada, D. ); Michaels, G.S.; Taylor, R. . Div. of Computer Research and Technology); Yoshida, Kaoru )

    1992-04-01

    GenoGraphics is a generic utility for constructing and querying one-dimensional linear plots. The outgrowth of a request from Dr. Cassandra Smith for a tool to facilitate her genome mapping research. GenoGraphics development has benefited from a continued collaboration with her. Written in Sun Microsystem's OpenWindows environment and the BTOL toolkit developed at Argonne National Laboratory. GenoGraphics provides an interactive, intuitive, graphical interface. Its features include: viewing multiple maps simultaneously, zooming, and querying by mouse clicking. By expediting plot generation, GenoGraphics gives the scientist more time to analyze data and a novel means for deducing conclusions.

  14. GenoGraphics for OpenWindows{trademark}

    SciTech Connect (OSTI)

    Hagstrom, R.; Overbeek, R.; Price, M.; Zawada, D.; Michaels, G.S.; Taylor, R.; Yoshida, Kaoru

    1992-04-01

    GenoGraphics is a generic utility for constructing and querying one-dimensional linear plots. The outgrowth of a request from Dr. Cassandra Smith for a tool to facilitate her genome mapping research. GenoGraphics development has benefited from a continued collaboration with her. Written in Sun Microsystem`s OpenWindows environment and the BTOL toolkit developed at Argonne National Laboratory. GenoGraphics provides an interactive, intuitive, graphical interface. Its features include: viewing multiple maps simultaneously, zooming, and querying by mouse clicking. By expediting plot generation, GenoGraphics gives the scientist more time to analyze data and a novel means for deducing conclusions.

  15. Extraction and analysis of neuron firing signals from deep cortical video microscopy

    SciTech Connect (OSTI)

    Kerekes, Ryan A; Blundon, Jay

    2014-01-01

    We introduce a method for extracting and analyzing neuronal activity time signals from video of the cortex of a live animal. The signals correspond to the firing activity of individual cortical neurons. Activity signals are based on the changing fluorescence of calcium indicators in the cells over time. We propose a cell segmentation method that relies on a user-specified center point, from which the signal extraction method proceeds. A stabilization approach is used to reduce tissue motion in the video. The extracted signal is then processed to flatten the baseline and detect action potentials. We show results from applying the method to a cortical video of a live mouse.

  16. OpenMSI: A Science Gateway to Sort Through Bio-Imaging's Big Datasets

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    OpenMSI: A Science Gateway to Sort Through Bio-Imaging's Big Datasets OpenMSI: A Science Gateway to Sort Through Bio-Imaging's Big Datasets August 27, 2013 Contact: Linda Vu, +1 510 495 2402, lvu@lbl.gov OpenMSINERSC.jpg This overlay of mass spectrometry images shows the spatial distribution of three different kind of lipids across a whole mouse cross-section. Lipids act as the structural components of cell membranes and are responsible for energy storage, among other things. Image credit:

  17. CHPRC1104-16_Rev06A

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Deep Vadose Zone Deep Vadose Zone Deep Vadose Zone Addressing contamination deep in the vadose zone Enter Please use your mouse to navigate CHPRC1104-16_Rev05_5-18-11 Contents • Where is the Hanford deep vadose zone? • What is the vadose zone? • How was it contaminated? • What are the risks? • Why is it so tough to clean up? • What is being done? • What’s next? • For more information Exit Contents Home Deep Vadose Zone Where is the Hanford deep vadose zone? At the Hanford Site in

  18. 2005 - 04 | Jefferson Lab

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    4 Apr 2005 Mon, 2005-04-25 14:00 Jefferson Lab's Detector Group builds small-animal imaging device for the German Cancer Research Center Wed, 2005-04-20 14:00 JLab, College of W&amp;M researchers study radiation blockers while conducting nuclear imaging of Iodine uptake in mouse tissues Wed, 2005-04-20 14:00 HAPPEx Results Hint at Strangely Magnetic Proton Wed, 2005-04-20 14:00 Is It or Isn't It? Pentaquark Debate Heats Up Mon, 2005-04-11 14:00 JLab, Hampton U. celebrate Einstein's love of

  19. Development of BAC libraries and integrated physical mapping of human chromosome 22 using BACs. Annual report, July 1994--June 1995

    SciTech Connect (OSTI)

    Kim, U.J.; Shizuya, Hiroaki; Simon, M.I.

    1995-12-31

    BACs and fosmids are stable, nonchimeric, and highly representative cloning systems. BACs maintain large-fragment genomic inserts (100 to 300 kb) that are easily prepared for most types of experiments, including DNA sequencing. The authors have improved the methods for generating BACs and developed extensive BAC libraries. They have constructed human BAC libraries with more than 175,000 clones from male fibroblast and sperm, and a mouse BAC library with more than 200,000 clones. The authors are currently expanding human library with the aim of achieving total 50X coverage human genomic library using sperm samples from anonymous donors.

  20. Ensemble Data Analysis ENvironment (EDEN)

    SciTech Connect (OSTI)

    Steed, Chad Allen

    2012-08-01

    The EDEN toolkit facilitates exploratory data analysis and visualization of global climate model simulation datasets. EDEN provides an interactive graphical user interface (GUI) that helps the user visually construct dynamic queries of the characteristically large climate datasets using temporal ranges, variable selections, and geographic areas of interest. EDEN reads the selected data into a multivariate visualization panel which features an extended implementation of parallel coordinates plots as well as interactive scatterplots. The user can query data in the visualization panel using mouse gestures to analyze different ranges of data. The visualization panel provides coordinated multiple views whereby selections made in one plot are propagated to the other plots.

  1. Using CAD software to simulate PV energy yield - The case of product integrated photovoltaic operated under indoor solar irradiation

    SciTech Connect (OSTI)

    Reich, N.H.; van Sark, W.G.J.H.M.; Turkenburg, W.C.; Sinke, W.C.

    2010-08-15

    In this paper, we show that photovoltaic (PV) energy yields can be simulated using standard rendering and ray-tracing features of Computer Aided Design (CAD) software. To this end, three-dimensional (3-D) sceneries are ray-traced in CAD. The PV power output is then modeled by translating irradiance intensity data of rendered images back into numerical data. To ensure accurate results, the solar irradiation data used as input is compared to numerical data obtained from rendered images, showing excellent agreement. As expected, also ray-tracing precision in the CAD software proves to be very high. To demonstrate PV energy yield simulations using this innovative concept, solar radiation time course data of a few days was modeled in 3-D to simulate distributions of irradiance incident on flat, single- and double-bend shapes and a PV powered computer mouse located on a window sill. Comparisons of measured to simulated PV output of the mouse show that also in practice, simulation accuracies can be very high. Theoretically, this concept has great potential, as it can be adapted to suit a wide range of solar energy applications, such as sun-tracking and concentrator systems, Building Integrated PV (BIPV) or Product Integrated PV (PIPV). However, graphical user interfaces of 'CAD-PV' software tools are not yet available. (author)

  2. A PAUF-neutralizing antibody targets both carcinoma and endothelial cells to impede pancreatic tumor progression and metastasis

    SciTech Connect (OSTI)

    Kim, Su Jin; Chang, Suhwan; Lee, Yangsoon; Kim, Na Young; Hwang, Yeonsil; Min, Hye Jin; Yoo, Kyung-Sook; Park, Eun Hye; Kim, Seokho; Chung, Young-Hwa; Park, Young Woo; Koh, Sang Seok

    2014-11-07

    Highlights: • PMAb83, a human monoclonal antibody against PAUF, impaired tumor progression in vivo. • PMAb83 attenuated aggressiveness of tumor cells and suppressed angiogenesis. • PMAb83 in combination with gemcitabine conferred improved survival of mouse model. - Abstract: Pancreatic adenocarcinoma up-regulated factor (PAUF) is expressed in pancreatic ductal adenocarcinoma (PDAC) and plays an important role in tumor progression and metastasis. Here we evaluate the anti-tumor efficacy of a human monoclonal antibody against PAUF, PMAb83, to provide a therapeutic intervention to treat the disease. PMAb83 reduced tumor growth and distant metastasis in orthotopically xenografted mice of human PDAC cells. PMAb83 treatment retarded proliferation along with weakened aggressiveness traits of the carcinoma cells. AKT/β-catenin signaling played a role in the carcinoma cell proliferation and the treated xenograft tumors exhibited reduced levels of β-catenin and cyclin D1. Moreover PMAb83 abrogated the PAUF-induced angiogenic responses of endothelial cells, reducing the density of CD31{sup +} vessels in the treated tumors. In combination with gemcitabine, PMAb83 conferred enhanced survival of xenografted mice by about twofold compared to gemcitabine alone. Taken together, our findings show that PMAb83 treatment decreases the aggressiveness of carcinoma cells and suppresses tumor vascularization, which culminates in mitigated tumor growth and metastasis with improved survival in PDAC mouse models.

  3. The influence of TRP53 in the dose response of radiation-induced apoptosis, DNA repair and genomic stability in murine haematopoietic cells

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Lemon, Jennifer A.; Taylor, Kristina; Verdecchia, Kyle; Phan, Nghi; Boreham, Douglas R.

    2014-01-01

    Apoptotic and DNA damage endpoints are frequently used as surrogate markers of cancer risk, and have been well-studied in the Trp53+/- mouse model. We report the effect of differing Trp53 gene status on the dose response of ionizing radiation exposures (0.01-2 Gy), with the unique perspective of determining if effects of gene status remain at extended time points. Here we report no difference in the dose response for radiation-induced DNA double-strand breaks in bone marrow and genomic instability (MN-RET levels) in peripheral blood, between wild-type (Trp53+/+) and heterozygous (Trp53+/-) mice. The dose response for Trp53+/+ mice showed higher initial levelsmore » of radiation-induced lymphocyte apoptosis relative to Trp53+/- between 0 and 1 Gy. Although this trend was observed up to 12 hours post-irradiation, both genotypes ultimately reached the same level of apoptosis at 14 hours, suggesting the importance of late-onset p53-independent apoptotic responses in this mouse model. Expected radiation-induced G1 cell cycle delay was observed in Trp53+/+ but not Trp53+/-. Although p53 has an important role in cancer risk, we have shown its influence on radiation dose response can be temporally variable. This research highlights the importance of caution when using haematopoietic endpoints as surrogates to extrapolate radiation-induced cancer risk estimation.« less

  4. Roles of miRNAs in microcystin-LR-induced Sertoli cell toxicity

    SciTech Connect (OSTI)

    Zhou, Yuan; Wang, Hui; Wang, Cong; Qiu, Xuefeng; Benson, Mikael; Yin, Xiaoqin; Xiang, Zou; Li, Dongmei; and others

    2015-08-15

    Microcystin (MC)-LR, a cyclic heptapeptide, is a potent reproductive system toxin. To understand the molecular mechanisms of MC-induced reproductive system cytotoxicity, we evaluated global changes of miRNA and mRNA expression in mouse Sertoli cells following MC-LR treatment. Our results revealed that the exposure to MC-LR resulted in an altered miRNA expression profile that might be responsible for the modulation of mRNA expression. Bio-functional analysis indicated that the altered genes were involved in specific cellular processes, including cell death and proliferation. Target gene analysis suggested that junction injury in Sertoli cells exposed to MC-LR might be mediated by miRNAs through the regulation of the Sertoli cell-Sertoli cell pathway. Collectively, these findings may enhance our understanding on the modes of action of MC-LR on mouse Sertoli cells as well as the molecular mechanisms underlying the toxicity of MC-LR on the male reproductive system. - Highlights: • miRNAs were altered in Sertoli cells exposed to MC-LR. • Alerted genes were involved in different cell functions including the cell morphology. • MC-LR adversely affected Sertoli cell junction formation through the regulating miRNAs.

  5. Poly(dimethylsiloxane) microchip-based immunoassay with multiple reaction zones: Toward on-chip multiplex detection platform

    SciTech Connect (OSTI)

    Shao, Guocheng; Wang, Jun; Li, Zhaohui; Saraf, Laxmikant V.; Wang, Wanjun; Lin, Yuehe

    2011-09-20

    In this work, a poly(dimethylsiloxane) (PDMS) microchip-based immuno-sensing platform with integrated pneumatic micro valves is described. The microchip was fabricated with multiple layer soft lithography technology. By controlling the activation status of corresponding valves, reagent flows in the microchannel network can be well manipulated so that immuno-reactions only take place at designated reaction zones (DRZs). Four DRZs are included in the prototype microchip. Since these DRZs are all isolated from each other by micro valves, cross contamination is prevented. Using the inner surface of the all-PDMS microchannel as immunoassay substrate, on-chip sandwich format solid phase immunoassay was performed to demonstrate the feasibility of this immuno-sensing platform. Mouse IgG and fluorescein isothiocyanate (FITC) were used as the model analyte and the signal reporter respectively. Only 10 ul sample is needed for the assay and low detection limit of 5 ng/ml (?33 pM) was achieved though low-cost polyclonal antibodies were used in our experiment for feasibility study only. The encouraging results from mouse IgG immunoassay proved the feasibility of our microchip design. With slight modification of the assay protocol, the same chip design can be used for multi-target detection and can provide a simple, cost-effective and integrated microchip solution for multiplex immunoassay applications.

  6. Characterization of evolutionary rates and constraints in three mammalian genomes

    SciTech Connect (OSTI)

    Cooper, Gregory M.; Brudno, Michael; Stone, Eric A.; Dubchak, Inna; Batzoglou, Serafim; Sidow, Arend

    2004-02-15

    We present an analysis of rates and patterns of microevolutionary phenomena that have shaped the human, mouse, and rat genomes since their last common ancestor. We find evidence for a shift in the mutational spectrum between the mouse and rat lineages, with the net effect being a relative increase in GC content in the rat genome. Our estimate for the neutral point substitution rate separating the two rodents is 0.196 substitutions per site, and 0.65 substitutions per site for the tree relating all three mammals. Small insertions and deletions of 1-10 bp in length (''microindels'') occur at approximately 5 percent of the point substitution rate. Inferred regional correlations in evolutionary rates between lineages and between types of sites support the idea that rates of evolution are influenced by local genomic or cell biological context. No substantial correlations between rates of point substitutions and rates of microindels are found, however, implying that the influences that affect these processes are distinct. Finally, we have identified those regions in the human genome that are evolving slowly, which are likely to include functional elements important to human biology. At least 5 percent of the human genome is under substantial constraint, most of which is noncoding.

  7. ChIP-seq Accurately Predicts Tissue-Specific Activity of Enhancers

    SciTech Connect (OSTI)

    Visel, Axel; Blow, Matthew J.; Li, Zirong; Zhang, Tao; Akiyama, Jennifer A.; Holt, Amy; Plajzer-Frick, Ingrid; Shoukry, Malak; Wright, Crystal; Chen, Feng; Afzal, Veena; Ren, Bing; Rubin, Edward M.; Pennacchio, Len A.

    2009-02-01

    A major yet unresolved quest in decoding the human genome is the identification of the regulatory sequences that control the spatial and temporal expression of genes. Distant-acting transcriptional enhancers are particularly challenging to uncover since they are scattered amongst the vast non-coding portion of the genome. Evolutionary sequence constraint can facilitate the discovery of enhancers, but fails to predict when and where they are active in vivo. Here, we performed chromatin immunoprecipitation with the enhancer-associated protein p300, followed by massively-parallel sequencing, to map several thousand in vivo binding sites of p300 in mouse embryonic forebrain, midbrain, and limb tissue. We tested 86 of these sequences in a transgenic mouse assay, which in nearly all cases revealed reproducible enhancer activity in those tissues predicted by p300 binding. Our results indicate that in vivo mapping of p300 binding is a highly accurate means for identifying enhancers and their associated activities and suggest that such datasets will be useful to study the role of tissue-specific enhancers in human biology and disease on a genome-wide scale.

  8. Mapping cis-Regulatory Domains in the Human Genome UsingMulti-Species Conservation of Synteny

    SciTech Connect (OSTI)

    Ahituv, Nadav; Prabhakar, Shyam; Poulin, Francis; Rubin, EdwardM.; Couronne, Olivier

    2005-06-13

    Our inability to associate distant regulatory elements with the genes that they regulate has largely precluded their examination for sequence alterations contributing to human disease. One major obstacle is the large genomic space surrounding targeted genes in which such elements could potentially reside. In order to delineate gene regulatory boundaries we used whole-genome human-mouse-chicken (HMC) and human-mouse-frog (HMF) multiple alignments to compile conserved blocks of synteny (CBS), under the hypothesis that these blocks have been kept intact throughout evolution at least in part by the requirement of regulatory elements to stay linked to the genes that they regulate. A total of 2,116 and 1,942 CBS>200 kb were assembled for HMC and HMF respectively, encompassing 1.53 and 0.86 Gb of human sequence. To support the existence of complex long-range regulatory domains within these CBS we analyzed the prevalence and distribution of chromosomal aberrations leading to position effects (disruption of a genes regulatory environment), observing a clear bias not only for mapping onto CBS but also for longer CBS size. Our results provide a genome wide data set characterizing the regulatory domains of genes and the conserved regulatory elements within them.

  9. Controlled-Resonant Surface Tapping-Mode Scanning Probe Electrospray Ionization Mass Spectrometry Imaging

    SciTech Connect (OSTI)

    Lorenz, Matthias; Ovchinnikova, Olga S; Kertesz, Vilmos; Van Berkel, Gary J

    2014-01-01

    This paper reports on the advancement of a controlled-resonance surface tapping-mode single capillary liquid junction extraction/ESI emitter for mass spectrometry imaging. The basic instrumental setup and the general operation of the system were discussed and optimized performance metrics were presented. The ability to spot sample, lane scan and chemically image in an automated and controlled fashion were demonstrated. Rapid, automated spot sampling was demonstrated for a variety of compound types including the cationic dye basic blue 7, the oligosaccharide cellopentaose, and the protein equine heart cytochrome c. The system was used for lane scanning and chemical imaging of the cationic dye crystal violet in inked lines on glass and for lipid distributions in mouse brain thin tissue sections. Imaging of the lipids in mouse brain tissue under optimized conditions provided a spatial resolution of approximately 35 m based on the ability to distinguish between features observed both in the optical and mass spectral chemical images. The sampling spatial resolution of this system was comparable to the best resolution that has been reported for other types of atmospheric pressure liquid extraction-based surface sampling/ionization techniques used for mass spectrometry imaging.

  10. Advanced slow-magic angle spinning probe for magnetic resonance imaging and spectroscopy

    DOE Patents [OSTI]

    Wind, Robert A.; Hu, Jian Zhi; Minard, Kevin R.; Rommereim, Donald N.

    2006-01-24

    The present invention relates to a probe and processes useful for magnetic resonance imaging and spectroscopy instruments. More particularly, the invention relates to a MR probe and processes for obtaining resolution enhancements of fluid objects, including live specimens, using an ultra-slow (magic angle) spinning (MAS) of the specimen combined with a modified phase-corrected magic angle turning (PHORMAT) pulse sequence. Proton NMR spectra were measured of the torso and the top part of the belly of a female BALBc mouse in a 2T field, while spinning the animal at a speed of 1.5 Hz. Results show that even in this relatively low field with PHORMAT, an isotropic spectrum is obtained with line widths that are a factor 4.6 smaller than those obtained in a stationary mouse. Resolution of 1H NMR metabolite spectra are thus significantly enhanced. Results indicate that PHORMAT has the potential to significantly increase the utility of 1H NMR spectroscopy for in vivo biochemical, biomedical and/or medical applications involving large-sized biological objects such as mice, rats and even humans within a hospital setting. For small-sized objects, including biological objects, such as excised tissues, organs, live bacterial cells, and biofilms, use of PASS at a spinning rate of 30 Hz and above is preferred.

  11. Matrix Effects in Biological Mass Spectrometry Imaging: Identification and Compensation

    SciTech Connect (OSTI)

    Lanekoff, Ingela T.; Stevens, Susan; Stenzel-Poore, Mary; Laskin, Julia

    2014-07-21

    Matrix effects in mass spectrometry imaging (MSI) may affect the observed molecular distribution in chemical and biological systems. In this study, we introduce an experimental approach that efficiently compensates for matrix effects in nanospray desorption electrospray ionization (nano-DESI) MSI without introducing any complexity into the experimental protocol. We demonstrate compensation for matrix effects in nano-DESI MSI of phosphatidylcholine (PC) in normal and ischemic mouse brain tissue by doping the nano-DESI solvent with PC standards. Specifically, we use mouse brain tissue of a middle cerebral artery occlusion (MCAO) stroke model with an ischemic region localized to one hemisphere of the brain. Due to similar suppression in ionization of endogenous PC molecules extracted from the tissue and PC standards added to the solvent, matrix effects are eliminated by normalizing the intensity of the sodium and potassium adducts of endogenous PC to the intensity of the corresponding adduct of the PC standard. This approach efficiently compensates for signal variations resulting from differences in the local concentrations of sodium and potassium in tissue sections and from the complexity of the extracted analyte mixture derived from local variations in molecular composition.

  12. Norathyriol Suppresses Skin Cancers Induced by Solar Ultraviolet Radiation by Targeting ERK Kinases

    SciTech Connect (OSTI)

    Li, Jixia; Malakhova, Margarita; Mottamal, Madhusoodanan; Reddy, Kanamata; Kurinov, Igor; Carper, Andria; Langfald, Alyssa; Oi, Naomi; Kim, Myoung Ok; Zhu, Feng; Sosa, Carlos P.; Zhou, Keyuan; Bode, Ann M.; Dong, Zigang

    2012-06-27

    Ultraviolet (UV) irradiation is the leading factor in the development of skin cancer, prompting great interest in chemopreventive agents for this disease. In this study, we report the discovery of norathyriol, a plant-derived chemopreventive compound identified through an in silico virtual screening of the Chinese Medicine Library. Norathyriol is a metabolite of mangiferin found in mango, Hypericum elegans, and Tripterospermum lanceolatum and is known to have anticancer activity. Mechanistic investigations determined that norathyriol acted as an inhibitor of extracellular signal-regulated kinase (ERK)1/2 activity to attenuate UVB-induced phosphorylation in mitogen-activated protein kinases signaling cascades. We confirmed the direct and specific binding of norathyriol with ERK2 through a cocrystal structural analysis. The xanthone moiety in norathyriol acted as an adenine mimetic to anchor the compound by hydrogen bonds to the hinge region of the protein ATP-binding site on ERK2. Norathyriol inhibited in vitro cell growth in mouse skin epidermal JB6 P+ cells at the level of G{sub 2}-M phase arrest. In mouse skin tumorigenesis assays, norathyriol significantly suppressed solar UV-induced skin carcinogenesis. Further analysis indicated that norathyriol mediates its chemopreventive activity by inhibiting the ERK-dependent activity of transcriptional factors AP-1 and NF-{kappa}B during UV-induced skin carcinogenesis. Taken together, our results identify norathyriol as a safe new chemopreventive agent that is highly effective against development of UV-induced skin cancer.

  13. Immunoglobulin λ Gene Rearrangement Can Precede κ Gene Rearrangement

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Berg, Jörg; Mcdowell, Mindy; Jäck, Hans-Martin; Wabl, Matthias

    1990-01-01

    Imore » mmunoglobulin genes are generated during differentiation of B lymphocytes by joining gene segments. A mouse pre-B cell contains a functional immunoglobulin heavy-chain gene, but no light-chain gene. Although there is only one heavy-chain locus, there are two lightchain loci: κ and λ .It has been reported that κ loci in the germ-line configuration are never (in man) or very rarely (in the mouse) present in cells with functionally rearranged λ -chain genes. Two explanations have been proposed to explain this: (a) the ordered rearrangement theory, which postulates that light-chain gene rearrangement in the pre-B cell is first attempted at the κ locus, and that only upon failure to produce a functional κ chain is there an attempt to rearrange the λ locus; and (b) the stochastic theory, which postulates that rearrangement at the λ locus proceeds at a rate that is intrinsically much slower than that at the κ locus. We show here that λ -chain genes are generated whether or not the κ locus has lost its germ-line arrangement, a result that is compatible only with the stochastic theory.« less

  14. Construction of genome-wide physical BAC contigs using mapped cDNA as probes: Toward an integrated BAC library resource for genome sequencing and analysis. Annual report, July 1995--January 1997

    SciTech Connect (OSTI)

    Mitchell, S.C.; Bocskai, D.; Cao, Y.

    1997-12-31

    The goal of human genome project is to characterize and sequence entire genomes of human and several model organisms, thus providing complete sets of information on the entire structure of transcribed, regulatory and other functional regions for these organisms. In the past years, a number of useful genetic and physical markers on human and mouse genomes have been made available along with the advent of BAC library resources for these organisms. The advances in technology and resource development made it feasible to efficiently construct genome-wide physical BAC contigs for human and other genomes. Currently, over 30,000 mapped STSs and 27,000 mapped Unigenes are available for human genome mapping. ESTs and cDNAs are excellent resources for building contig maps for two reasons. Firstly, they exist in two alternative forms--as both sequence information for PCR primer pairs, and cDoreen genomic libraries efficiently for large number of DNA probes by combining over 100 cDNA probes in each hybridization. Second, the linkage and order of genes are rather conserved among human, mouse and other model organisms. Therefore, gene markers have advantages over random anonymous STSs in building maps for comparative genomic studies.

  15. H2-M polymorphism in mice susceptible to collagen-induced arthritis involves the peptide binding groove

    SciTech Connect (OSTI)

    Walter, W.; Loos, M.; Maeurer, M.J.

    1996-12-31

    The ability to develop type II collagen (CII)-induced arthritis (CIA) in mice is associated with the major histocompatibility I-A gene and with as yet poorly defined regulatory molecules of the major histocompatibility complex (MHC) class II antigen processing and presentation pathway. H2-M molecules are thought to be involved in the loading of antigenic peptides into the MHC class II binding cleft. We sequenced H2-Ma, H2-Mb1, and H2-Mb2 genes from CIA-susceptible and -resistant mouse strains and identified four different Ma and Mb2 alleles, and three different Mb1 alleles defined by polymorphic residues within the predicted peptide binding groove. Most CIA-resistant mouse strains share common Ma, Mb1, and Mb2 alleles. In contrast, H2-M alleles designated Ma-III, Ma-IV, Mb1-III, and Mb2-IV could be exclusively identified in the CIA-susceptible H2{sup r} and H2{sup q} haplotypes, suggesting that allelic H2-M molecules may modulate the composition of different CII peptides loaded onto MHC class II molecules, presumably presenting {open_quotes}arthritogenic{close_quotes} epitopes to T lymphocytes. 42 refs., 4 figs., 3 tabs.

  16. Critical role of iodination for T cell recognition of thyroglobulin in experimental murine thyroid autoimmunity

    SciTech Connect (OSTI)

    Champion, B.R.; Rayner, D.C.; Byfield, P.G.H.; Page, K.R.; Chan, C.T.J.; Roitt, I.M.

    1987-12-01

    The authors have used two clonotypically distinct thyroglobulin (Tg)-specific, I-A/sup k/-restricted monoclonal T cell populations to investigate the role of thyroid peroxidase-catalyzed iodination in Tg recognition by autorreactive T cells. The results showed that these T cells could recognize Tg only it was sufficiently iodinated. Unlike normal mouse Tg, noniodinated mouse Tg was unable to induce significant thyroid lesions but could trigger the production of Tg autoantibodies. In these experiments, the importance of T cell recognition of iodination-related epitopes was emphasized by the inability of serum antibodies to distinguish Tg on the basis of iodine content, whether they were induced with normal or noniodinated Tg. Therefore, thyroid peroxidase-dependent modification of Tg would appear to be central to its recognition by autoreactive T cells and hence its capacity to induce autoimmune thyroid lesions. Proliferative responses of the Tg-specific T cell clone was assessed by the incorporation of (/sup 125/I) deoxyuridine. Anti-Tg antibody activity was determined by radioimmunoassay.

  17. Online adaptation and verification of VMAT

    SciTech Connect (OSTI)

    Crijns, Wouter; Defraene, Gilles; Depuydt, Tom; Haustermans, Karin; Van Herck, Hans; Maes, Frederik; Van den Heuvel, Frank

    2015-07-15

    Purpose: This work presents a method for fast volumetric modulated arc therapy (VMAT) adaptation in response to interfraction anatomical variations. Additionally, plan parameters extracted from the adapted plans are used to verify the quality of these plans. The methods were tested as a prostate class solution and compared to replanning and to their current clinical practice. Methods: The proposed VMAT adaptation is an extension of their previous intensity modulated radiotherapy (IMRT) adaptation. It follows a direct (forward) planning approach: the multileaf collimator (MLC) apertures are corrected in the beam’s eye view (BEV) and the monitor units (MUs) are corrected using point dose calculations. All MLC and MU corrections are driven by the positions of four fiducial points only, without need for a full contour set. Quality assurance (QA) of the adapted plans is performed using plan parameters that can be calculated online and that have a relation to the delivered dose or the plan quality. Five potential parameters are studied for this purpose: the number of MU, the equivalent field size (EqFS), the modulation complexity score (MCS), and the components of the MCS: the aperture area variability (AAV) and the leaf sequence variability (LSV). The full adaptation and its separate steps were evaluated in simulation experiments involving a prostate phantom subjected to various interfraction transformations. The efficacy of the current VMAT adaptation was scored by target mean dose (CTV{sub mean}), conformity (CI{sub 95%}), tumor control probability (TCP), and normal tissue complication probability (NTCP). The impact of the adaptation on the plan parameters (QA) was assessed by comparison with prediction intervals (PI) derived from a statistical model of the typical variation of these parameters in a population of VMAT prostate plans (n = 63). These prediction intervals are the adaptation equivalent of the tolerance tables for couch shifts in the current clinical practice. Results: The proposed adaptation of a two-arc VMAT plan resulted in the intended CTV{sub mean} (Δ ≤ 3%) and TCP (ΔTCP ≤ 0.001). Moreover, the method assures the intended CI{sub 95%} (Δ ≤ 11%) resulting in lowered rectal NTCP for all cases. Compared to replanning, their adaptation is faster (13 s vs 10 min) and more intuitive. Compared to the current clinical practice, it has a better protection of the healthy tissue. Compared to IMRT, VMAT is more robust to anatomical variations, but it is also less sensitive to the different correction steps. The observed variations of the plan parameters in their database included a linear dependence on the date of treatment planning and on the target radius. The MCS is not retained as QA metric due to a contrasting behavior of its components (LSV and AAV). If three out of four plan parameters (MU, EqFS, AAV, and LSV) need to lie inside a 50% prediction interval (3/4—50%PI), all adapted plans will be accepted. In contrast, all replanned plans do not meet this loose criterion, mainly because they have no connection to the initially optimized and verified plan. Conclusions: A direct (forward) VMAT adaptation performs equally well as (inverse) replanning but is faster and can be extended to real-time adaptation. The prediction intervals for the machine parameters are equivalent to the tolerance tables for couch shifts in the current clinical practice. A 3/4—50%PI QA criterion accepts all the adapted plans but rejects all the replanned plans.

  18. Tuning the DARHT Axis-II linear induction accelerator focusing

    SciTech Connect (OSTI)

    Ekdahl, Carl A.

    2012-04-24

    Flash radiography of large hydrodynamic experiments driven by high explosives is a well-known diagnostic technique in use at many laboratories, and the Dual-Axis Radiography for Hydrodynamic Testing (DARHT) facility at Los Alamos produces flash radiographs of large hydrodynamic experiments. Two linear induction accelerators (LIAs) make the bremsstrahlung radiographic source spots for orthogonal views of each test. The 2-kA, 20-MeV Axis-I LIA creates a single 60-ns radiography pulse. The 1.7-kA, 16.5-MeV Axis-II LIA creates up to four radiography pulses by kicking them out of a longer pulse that has a 1.6-{mu}s flattop. The Axis-II injector, LIA, kicker, and downstream transport (DST) to the bremsstrahlung converter are described. Adjusting the magnetic focusing and steering elements to optimize the electron-beam transport through an LIA is often called 'tuning.' As in all high-current LIAs, the focusing field is designed to be as close to that of the ideal continuous solenoid as physically possible. In ideal continuous solenoidal transport a smoothly varying beam size can easily be found for which radial forces balance, and the beam is said to be 'matched' to the focusing field. A 'mismatched' beam exhibits unwanted oscillations in size, which are a source of free energy that contributes to emittance growth. This is undesirable, because in the absence of beam-target effects, the radiographic spot size is proportional to the emittance. Tuning the Axis-II LIA is done in two steps. First, the solenoidal focusing elements are set to values designed to provide a matched beam with little or no envelope oscillations, and little or no beam-breakup (BBU) instability growth. Then, steering elements are adjusted to minimize the motion of the centroid of a well-centered beam at the LIA exit. This article only describes the design of the tune for the focusing solenoids. The DARHT Axis-II LIA was required to be re-tuned after installing an accelerator cell to replace a failed solenoid in March of 2012. We took advantage of this opportunity to improve the design of the focusing tune with better models of the remaining partially failed solenoids, better estimates of beam initial conditions, and better values for pulsed-power voltages. As with all previous tunes for Axis-II, this one incorporates measures to mitigate beam-breakup (BBU) instability, image displacement instability (IDI), corkscrew (sweep), and emittance growth. Section II covers the general approach to of design of focusing solenoid tunes for the DARHT Axis-2 LIA. Section III explains the specific requirements and simulations needed to design the tune for the injector, which includes the thermionic electron source, diode, and six induction cells. Section IV explains the requirements and simulations for tuning the main accelerator, which consists of 68 induction cells. Finally, Section V explores sensitivity of the tune to deviations of parameters from nominal, random variations, and uncertainties in values. Four appendices list solenoid settings for this new tune, discuss comparisons of different simulation codes, show halo formation in mismatched beams, and present a brief discussion of the beam envelope equation, which is the heart of the method used to design LIA solenoid tunes.

  19. Quality assurance for online adapted treatment plans: Benchmarking and delivery monitoring simulation

    SciTech Connect (OSTI)

    Li, Taoran Wu, Qiuwen; Yang, Yun; Rodrigues, Anna; Yin, Fang-Fang; Jackie Wu, Q.

    2015-01-15

    Purpose: An important challenge facing online adaptive radiation therapy is the development of feasible and efficient quality assurance (QA). This project aimed to validate the deliverability of online adapted plans and develop a proof-of-concept online delivery monitoring system for online adaptive radiation therapy QA. Methods: The first part of this project benchmarked automatically online adapted prostate treatment plans using traditional portal dosimetry IMRT QA. The portal dosimetry QA results of online adapted plans were compared to original (unadapted) plans as well as randomly selected prostate IMRT plans from our clinic. In the second part, an online delivery monitoring system was designed and validated via a simulated treatment with intentional multileaf collimator (MLC) errors. This system was based on inputs from the dynamic machine information (DMI), which continuously reports actual MLC positions and machine monitor units (MUs) at intervals of 50 ms or less during delivery. Based on the DMI, the system performed two levels of monitoring/verification during the delivery: (1) dynamic monitoring of cumulative fluence errors resulting from leaf position deviations and visualization using fluence error maps (FEMs); and (2) verification of MLC positions against the treatment plan for potential errors in MLC motion and data transfer at each control point. Validation of the online delivery monitoring system was performed by introducing intentional systematic MLC errors (ranging from 0.5 to 2 mm) to the DMI files for both leaf banks. These DMI files were analyzed by the proposed system to evaluate the systems performance in quantifying errors and revealing the source of errors, as well as to understand patterns in the FEMs. In addition, FEMs from 210 actual prostate IMRT beams were analyzed using the proposed system to further validate its ability to catch and identify errors, as well as establish error magnitude baselines for prostate IMRT delivery. Results: Online adapted plans were found to have similar delivery accuracy in comparison to clinical IMRT plans when validated with portal dosimetry IMRT QA. FEMs for the simulated deliveries with intentional MLC errors exhibited distinct patterns for different MLC error magnitudes and directions, indicating that the proposed delivery monitoring system is highly specific in detecting the source of errors. Implementing the proposed QA system for online adapted plans revealed excellent delivery accuracy: over 99% of leaf position differences were within 0.5 mm, and >99% of pixels in the FEMs had fluence errors within 0.5 MU. Patterns present in the FEMs and MLC control point analysis for actual patient cases agreed with the error pattern analysis results, further validating the systems ability to reveal and differentiate MLC deviations. Calculation of the fluence map based on the DMI was performed within 2 ms after receiving each DMI input. Conclusions: The proposed online delivery monitoring system requires minimal additional resources and time commitment to the current clinical workflow while still maintaining high sensitivity to leaf position errors and specificity to error types. The presented online delivery monitoring system therefore represents a promising QA system candidate for online adaptive radiation therapy.

  20. A novel synthetic derivative of melatonin, 5-hydroxy-2’-isobutyl-streptochlorin (HIS), inhibits inflammatory responses via regulation of TRIF-dependent signaling and inflammasome activation

    SciTech Connect (OSTI)

    Shim, Do-Wan; Shin, Hee Jae; Han, Ji-Won; Ji, Young-Eun; Jang, Cheol-Hun; Koppula, Sushruta; Kang, Tae-Bong; Lee, Kwang-Ho

    2015-04-15

    Melatonin is substantially reported to possess anti-inflammatory properties. In the present study, we synthesized a novel melatonin derivative, 5-hydroxy-2′-isobutyl-streptochlorin (HIS), which displayed superior anti-inflammatory properties to its parent compound. Further, we explored its underlying mechanisms in cellular and experimental animal models. Lipopolysaccharide was used to induce in vitro inflammatory responses in RAW 264.7 macrophages. LPS-primed macrophages were pulsed with biologically unrelated toxic molecules to evaluate the role of HIS on inflammasome activation. In vivo verifications were carried out using acute lung injury (ALI) and Escherichia coli-induced septic shock mouse models. HIS inhibited the production of proinflammatory mediators and cytokines such as nitric oxide, cyclooxygenase 2, IL-1β, IL-6 and TNF-α in LPS-stimulated RAW 264.7 macrophages. HIS suppressed the infiltration of immune cells into the lung and the production of pro-inflammatory cytokines such as IL-6 and TNF-α in broncho-alveolar lavage fluid in the ALI mouse model. Mechanistic studies revealed that the inhibitory effects of HIS were mediated through the regulation of the TIR domain-containing, adaptor-inducing, interferon-β (TRIF)-dependent signaling pathway from toll-like receptors. Further, HIS attenuated IL-1β secretion via the inhibition of NLRP3 inflammasome activation independent of mitochondrial ROS production. Furthermore, HIS suppressed IL-1β, IL-6 and interferon-β production in peritoneal lavage in the Escherichia coli-induced sepsis mouse model. In conclusion, HIS exerted potent anti-inflammatory effects via the regulation of TRIF-dependent signaling and inflammasome activation. Notably, the superior anti-inflammatory properties of this derivative compared with its parent compound could be a promising lead for treating various inflammatory-mediated diseases. - Highlights: • Νovel compound, 5-hydroxy-2′-isobutyl-streptochlorin (HIS) was synthesized. • HIS inhibited TRIF-dependent signaling induced by LPS stimulation. • HIS inhibited NLRP3 inflammasome activation. • HIS inhibited acute lung injury and Escherichia coli-induced septic shock.

  1. MO-G-BRF-09: Investigating Magnetic Field Dose Effects in Mice: A Monte Carlo Study

    SciTech Connect (OSTI)

    Rubinstein, A; Guindani, M; Followill, D; Melancon, A; Hazle, J; Court, L

    2014-06-15

    Purpose: In MRI-linac treatments, radiation dose distributions are affected by magnetic fields, especially at high-density/low-density interfaces. Radiobiological consequences of magnetic field dose effects are presently unknown; therefore, preclinical studies are needed to ensure the safe clinical use of MRI-linacs. This study investigates the optimal combination of beam energy and magnetic field strength needed for preclinical murine studies. Methods: The Monte Carlo code MCNP6 was used to simulate the effects of a magnetic field when irradiating a mouse-sized lung phantom with a 1.0cmx1.0cm photon beam. Magnetic field effects were examined using various beam energies (225kVp, 662keV[Cs-137], and 1.25MeV[Co-60]) and magnetic field strengths (0.75T, 1.5T, and 3T). The resulting dose distributions were compared to Monte Carlo results for humans with various field sizes and patient geometries using a 6MV/1.5T MRI-linac. Results: In human simulations, the addition of a 1.5T magnetic field caused an average dose increase of 49% (range:36%60%) to lung at the soft tissue-to-lung interface and an average dose decrease of 30% (range:25%36%) at the lung-to-soft tissue interface. In mouse simulations, the magnetic fields had no effect on the 225kVp dose distribution. The dose increases for the Cs-137 beam were 12%, 33%, and 49% for 0.75T, 1.5T, and 3.0T magnetic fields, respectively while the dose decreases were 7%, 23%, and 33%. For the Co-60 beam, the dose increases were 14%, 45%, and 41%, and the dose decreases were 18%, 35%, and 35%. Conclusion: The magnetic field dose effects observed in mouse phantoms using a Co-60 beam with 1.5T or 3T fields and a Cs-137 beam with a 3T field compare well with those seen in simulated human treatments with an MRI-linac. These irradiator/magnet combinations are suitable for preclinical studies investigating potential biological effects of delivering radiation therapy in the presence of a magnetic field. Partially funded by Elekta.

  2. Modulation of keratinocyte expression of antioxidants by 4-hydroxynonenal, a lipid peroxidation end product

    SciTech Connect (OSTI)

    Zheng, Ruijin; Heck, Diane E.; Mishin, Vladimir; Black, Adrienne T.; Shakarjian, Michael P.; Kong, Ah-Ng Tony; Laskin, Debra L.; Laskin, Jeffrey D.

    2014-03-01

    4-Hydroxynonenal (4-HNE) is a lipid peroxidation end product generated in response to oxidative stress in the skin. Keratinocytes contain an array of antioxidant enzymes which protect against oxidative stress. In these studies, we characterized 4-HNE-induced changes in antioxidant expression in mouse keratinocytes. Treatment of primary mouse keratinocytes and PAM 212 keratinocytes with 4-HNE increased mRNA expression for heme oxygenase-1 (HO-1), catalase, NADPH:quinone oxidoreductase (NQO1) and glutathione S-transferase (GST) A1-2, GSTA3 and GSTA4. In both cell types, HO-1 was the most sensitive, increasing 8698 fold within 6 h. Further characterization of the effects of 4-HNE on HO-1 demonstrated concentration- and time-dependent increases in mRNA and protein expression which were maximum after 6 h with 30 ?M. 4-HNE stimulated keratinocyte Erk1/2, JNK and p38 MAP kinases, as well as PI3 kinase. Inhibition of these enzymes suppressed 4-HNE-induced HO-1 mRNA and protein expression. 4-HNE also activated Nrf2 by inducing its translocation to the nucleus. 4-HNE was markedly less effective in inducing HO-1 mRNA and protein in keratinocytes from Nrf2 ?/? mice, when compared to wild type mice, indicating that Nrf2 also regulates 4-HNE-induced signaling. Western blot analysis of caveolar membrane fractions isolated by sucrose density centrifugation demonstrated that 4-HNE-induced HO-1 is localized in keratinocyte caveolae. Treatment of the cells with methyl-?-cyclodextrin, which disrupts caveolar structure, suppressed 4-HNE-induced HO-1. These findings indicate that 4-HNE modulates expression of antioxidant enzymes in keratinocytes, and that this can occur by different mechanisms. Changes in expression of keratinocyte antioxidants may be important in protecting the skin from oxidative stress. - Highlights: Lipid peroxidation generates 4-hydroxynonenal, a reactive aldehyde. 4-HNE induces antioxidant proteins in mouse keratinocytes. Induction of antioxidant proteins is regulated via MAP kinases, Nrf2 and caveolae. 4-HNE is an effective signaling molecule in keratinocytes.

  3. Predicting carcinogenicity of diverse chemicals using probabilistic neural network modeling approaches

    SciTech Connect (OSTI)

    Singh, Kunwar P.; Gupta, Shikha; Rai, Premanjali

    2013-10-15

    Robust global models capable of discriminating positive and non-positive carcinogens; and predicting carcinogenic potency of chemicals in rodents were developed. The dataset of 834 structurally diverse chemicals extracted from Carcinogenic Potency Database (CPDB) was used which contained 466 positive and 368 non-positive carcinogens. Twelve non-quantum mechanical molecular descriptors were derived. Structural diversity of the chemicals and nonlinearity in the data were evaluated using Tanimoto similarity index and BrockDechertScheinkman statistics. Probabilistic neural network (PNN) and generalized regression neural network (GRNN) models were constructed for classification and function optimization problems using the carcinogenicity end point in rat. Validation of the models was performed using the internal and external procedures employing a wide series of statistical checks. PNN constructed using five descriptors rendered classification accuracy of 92.09% in complete rat data. The PNN model rendered classification accuracies of 91.77%, 80.70% and 92.08% in mouse, hamster and pesticide data, respectively. The GRNN constructed with nine descriptors yielded correlation coefficient of 0.896 between the measured and predicted carcinogenic potency with mean squared error (MSE) of 0.44 in complete rat data. The rat carcinogenicity model (GRNN) applied to the mouse and hamster data yielded correlation coefficient and MSE of 0.758, 0.71 and 0.760, 0.46, respectively. The results suggest for wide applicability of the inter-species models in predicting carcinogenic potency of chemicals. Both the PNN and GRNN (inter-species) models constructed here can be useful tools in predicting the carcinogenicity of new chemicals for regulatory purposes. - Graphical abstract: Figure (a) shows classification accuracies (positive and non-positive carcinogens) in rat, mouse, hamster, and pesticide data yielded by optimal PNN model. Figure (b) shows generalization and predictive abilities of the interspecies GRNN model to predict the carcinogenic potency of diverse chemicals. - Highlights: Global robust models constructed for carcinogenicity prediction of diverse chemicals. Tanimoto/BDS test revealed structural diversity of chemicals and nonlinearity in data. PNN/GRNN successfully predicted carcinogenicity/carcinogenic potency of chemicals. Developed interspecies PNN/GRNN models for carcinogenicity prediction. Proposed models can be used as tool to predict carcinogenicity of new chemicals.

  4. Development of Biomarkers for Chronic Beryllium Disease in Mice

    SciTech Connect (OSTI)

    Gordon, Terry

    2013-01-25

    Beryllium is a strategic metal, indispensable for national defense programs in aerospace, telecommunications, electronics, and weaponry. Exposure to beryllium is an extensively documented occupational hazard that causes irreversible, debilitating granulomatous lung disease in as much as 3 - 5% of exposed workers. Mechanistic research on beryllium exposure-disease relationships has been severely limited by a general lack of a sufficient CBD animal model. We have now developed and tested an animal model which can be used for dissecting dose-response relationships and pathogenic mechanisms and for testing new diagnostic and treatment paradigms. We have created 3 strains of transgenic mice in which the human antigen-presenting moiety, HLA-DP, was inserted into the mouse genome. Each mouse strain contains HLA-DPB1 alleles that confer different magnitude of risk for chronic beryllium disease (CBD): HLA-DPB1*0401 (odds ratio = 0.2), HLA-DPB1*0201 (odds ratio = 15), HLA-DPB1*1701 (odds ratio = 240). Our preliminary work has demonstrated that the *1701 allele, as predicted by human studies, results in the greatest degree of sensitization in a mouse ear swelling test. We have also completed dose-response experiments examining beryllium-induced lung granulomas and identified susceptible and resistant inbred strains of mice (without the human transgenes) as well as quantitative trait loci that may contain gene(s) that modify the immune response to beryllium. In this grant application, we propose to use the transgenic and ?normal inbred strains of mice to identify biomarkers for the progression of beryllium sensitization and CBD. To achieve this goal, we propose to compare the sensitivity and accuracy of the lymphocyte proliferation test (blood and bronchoalveolar lavage fluid) with the ELISPOT test in the three HLA-DP transgenic mice strains throughout a 6 month treatment with beryllium particles. Because of the availability of high-throughput proteomics, we will also identify changes in potential protein biomarkers in beryllium-treated mice. We will correlate these findings with the ability of the transgenic mice to develop a beryllium-specific adaptive immune response in blood and bronchoalveolar lavage (BAL) fluid. We will also determine whether beryllium-responsive CD4+ T cells in blood and BAL correlate with the onset of granuloma formation. Thus, we will provide the scientific community with biomarkers of sensitization and disease progression for CBD. These biomarkers will serve as critical tools for development of improved industrial hygiene and therapeutic interventions.

  5. Sulforaphane is not an effective antagonist of the human pregnane X-receptor in vivo

    SciTech Connect (OSTI)

    Poulton, Emma Jane; Department of Environmental and Occupational Health Sciences, University of Washington ; Levy, Lisa; Lampe, Johanna W.; Public Health Sciences Division, Fred Hutchinson Cancer Research Center ; Shen, Danny D.; Department of Pharmaceutics, University of Washington ; Tracy, Julia; Department of Environmental and Occupational Health Sciences, University of Washington ; Shuhart, Margaret C.; Thummel, Kenneth E.; Department of Pharmaceutics, University of Washington ; Eaton, David L.

    2013-01-01

    Sulforaphane (SFN), is an effective in vitro antagonist of ligand activation of the human pregnane and xenobiotic receptor (PXR). PXR mediated CYP3A4 up-regulation is implicated in adverse drug-drug interactions making identification of small molecule antagonists a desirable therapeutic goal. SFN is not an antagonist to mouse or rat PXR in vitro; thus, normal rodent species are not suitable as in vivo models for human response. To evaluate whether SFN can effectively antagonize ligand activation of human PXR in vivo, a three-armed, randomized, crossover trial was conducted with 24 healthy adults. The potent PXR ligand rifampicin (300 mg/d) was given alone for 7 days in arm 1, or in daily combination with 450 ?mol SFN (Broccoli Sprout extract) in arm 2; SFN was given alone in arm 3. Midazolam as an in vivo phenotype marker of CYP3A was administered before and after each treatment arm. Rifampicin alone decreased midazolam AUC by 70%, indicative of the expected increase in CYP3A4 activity. Co-treatment with SFN did not reduce CYP3A4 induction. Treatment with SFN alone also did not affect CYP3A4 activity in the cohort as a whole, although in the subset with the highest basal CYP3A4 activity there was a statistically significant increase in midazolam AUC (i.e., decrease in CYP3A4 activity). A parallel study in humanized PXR mice yielded similar results. The parallel effects of SFN between humanized PXR mice and human subjects demonstrate the predictive value of humanized mouse models in situations where species differences in ligand-receptor interactions preclude the use of a native mouse model for studying human ligand-receptor pharmacology. -- Highlights: ? The effects of SFN on PXR mediated CYP3A4 induction in humanized PXR mice and humans were examined. ? SFN had no effect on rifampicin mediated CYP3A4 induction in humans or humanized mice. ? SFN had a modest effect on basal CYP3A4 activity among subjects with higher baseline activity. ? Humanized PXR mice were generally predictive of the in vivo human response.

  6. Detection of genotoxic and non-genotoxic carcinogens in Xpc{sup ?/?}p53{sup +/?} mice

    SciTech Connect (OSTI)

    Melis, Joost P.M.; Leiden University Medical Center, Department of Toxicogenetics, Leiden ; Speksnijder, Ewoud N.; Kuiper, Raoul V.; Dutch Molecular Pathology Center, Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Utrecht ; Salvatori, Daniela C.F.; Schaap, Mirjam M.; Leiden University Medical Center, Department of Toxicogenetics, Leiden ; Maas, Saskia; Robinson, Joke; Verhoef, Aart; Benthem, Jan van; Luijten, Mirjam; Steeg, Harry van

    2013-01-15

    An accurate assessment of the carcinogenic potential of chemicals and pharmaceutical drugs is essential to protect humans and the environment. Therefore, substances are extensively tested before they are marketed to the public. Currently, the rodent two-year bioassay is still routinely used to assess the carcinogenic potential of substances. However, over time it has become clear that this assay yields false positive results and also has several economic and ethical drawbacks including the use of large numbers of animals, the long duration, and the high cost. The need for a suitable alternative assay is therefore high. Previously, we have proposed the Xpa*p53 mouse model as a very suitable alternative to the two-year bioassay. We now show that the Xpc*p53 mouse model preserves all the beneficial traits of the Xpa*p53 model for sub-chronic carcinogen identification and can identify both genotoxic and non-genotoxic carcinogens. Moreover, Xpc*p53 mice appear to be more responsive than Xpa*p53 mice towards several genotoxic and non-genotoxic carcinogens. Furthermore, Xpc*p53 mice are far less sensitive than Xpa*p53 mice for the toxic activity of DNA damaging agents and as such clearly respond in a similar way as wild type mice do. These advantageous traits of the Xpc*p53 model make it a better alternative for in vivo carcinogen testing than Xpa*p53. This pilot study suggests that Xpc*p53 mice are suited for routine sub-chronic testing of both genotoxic and non-genotoxic carcinogens and as such represent a suitable alternative to possibly replace the murine life time cancer bioassay. Highlights: ? The Xpc*p53 mouse model is able to identify genotoxic and non-genotoxic carcinogens. ? Time, animals and cost can be significantly reduced compared to the 2-year bioassay. ? Xpc*p53 mice are more advantageous for carcinogen identification than Xpa*p53 mice. ? Xpc*p53 mice exhibit a wild type response upon exposure to genotoxicants.

  7. Environmental Survey Report for ORNL: Small Mammal Abundance and Distribution Survey Oak Ridge National Environmental Research Park 2009 - 2010

    SciTech Connect (OSTI)

    Giffen, Neil R; Reasor, R. Scott; Campbell, Claire L.

    2009-12-01

    This report summarizes a 1-year small mammal biodiversity survey conducted on the Oak Ridge National Environmental Research Park (OR Research Park). The task was implemented through the Oak Ridge National Laboratory (ORNL) Natural Resources Management Program and included researchers from the ORNL Environmental Sciences Division, interns in the Oak Ridge Institute for Science and Education Higher Education Research Experiences Program, and ORNL Environmental Protection Services staff. Eight sites were surveyed reservation wide. The survey was conducted in an effort to determine species abundance and diversity of small mammal populations throughout the reservation and to continue the historical inventory of small mammal presence for biodiversity records. This data collection effort was in support of the approved Wildlife Management Plan for the Oak Ridge Reservation, a major goal of which is to maintain and enhance wildlife biodiversity on the Reservation. Three of the sites (Poplar Creek, McNew Hollow, and Deer Check Station Field) were previously surveyed during a major natural resources inventory conducted in 1996. Five new sites were included in this study: Bearden Creek, Rainy Knob (Natural Area 21), Gum Hollow, White Oak Creek and Melton Branch. The 2009-2010 small mammal surveys were conducted from June 2009 to July 2010 on the Oak Ridge National Environmental Research Park (OR Research Park). The survey had two main goals: (1) to determine species abundance and diversity and (2) to update historical records on the OR Research Park. The park is located on the Department of Energy-owned Oak Ridge Reservation, which encompasses 13,580 ha. The primary focus of the study was riparian zones. In addition to small mammal sampling, vegetation and coarse woody debris samples were taken at certain sites to determine any correlations between habitat and species presence. During the survey all specimens were captured and released using live trapping techniques including Sherman and pitfall traps. In total 227 small mammals representing nine species were captured during the course of the study. The most common species found in the study was the white-footed mouse (Peromyscus leucopus). The least common species found were the deer mouse (Peromyscus maniculatus), meadow jumping mouse (Zapus hudsonius), woodland vole (Microtus pinetorum), and northern short-tailed shrew (Blarina brevicauda).

  8. Threatened and Endangered Species Habitat Management Plan for Los Alamos National Laboratory

    SciTech Connect (OSTI)

    Keller, David Charles; Hathcock, Charles Dean

    2015-11-17

    Los Alamos National Laboratory’s (LANL) Threatened and Endangered Species Habitat Management Plan (HMP) fulfills a commitment made to the U.S. Department of Energy (DOE) in the “Final Environmental Impact Statement for the Dual-Axis Radiographic Hydrodynamic Test Facility Mitigation Action Plan” (DOE 1996). The HMP received concurrence from the U.S. Fish and Wildlife Service (USFWS) in 1999 (USFWS consultation numbers 2-22-98-I-336 and 2-22-95-I-108). This 2015 update retains the management guidelines from the 1999 HMP for listed species, updates some descriptive information, and adds the New Mexico Meadow Jumping Mouse (Zapus hudsonius luteus) and Yellow-billed Cuckoo (Coccyzus americanus) which were federally listed in 2014 (Keller 2015: USFWS consultation number 02ENNM00- 2015-I-0538).

  9. An Efficient Algorithm for Mapping Imaging Data to 3D Unstructured Grids in Computational Biomechanics

    SciTech Connect (OSTI)

    Einstein, Daniel R.; Kuprat, Andrew P.; Jiao, Xiangmin; Carson, James P.; Einstein, David M.; Corley, Richard A.; Jacob, Rick E.

    2013-01-01

    Geometries for organ scale and multiscale simulations of organ function are now routinely derived from imaging data. However, medical images may also contain spatially heterogeneous information other than geometry that are relevant to such simulations either as initial conditions or in the form of model parameters. In this manuscript, we present an algorithm for the efficient and robust mapping of such data to imaging based unstructured polyhedral grids in parallel. We then illustrate the application of our mapping algorithm to three different mapping problems: 1) the mapping of MRI diffusion tensor data to an unstuctured ventricular grid; 2) the mapping of serial cyro-section histology data to an unstructured mouse brain grid; and 3) the mapping of CT-derived volumetric strain data to an unstructured multiscale lung grid. Execution times and parallel performance are reported for each case.

  10. Six degree of freedom sensor

    DOE Patents [OSTI]

    Vann, Charles S.

    1999-01-01

    This small, non-contact optical sensor increases the capability and flexibility of computer controlled machines by detecting its relative position to a workpiece in all six degrees of freedom (DOF). At a fraction of the cost, it is over 200 times faster and up to 25 times more accurate than competing 3-DOF sensors. Applications range from flexible manufacturing to a 6-DOF mouse for computers. Until now, highly agile and accurate machines have been limited by their inability to adjust to changes in their tasks. By enabling them to sense all six degrees of position, these machines can now adapt to new and complicated tasks without human intervention or delay--simplifying production, reducing costs, and enhancing the value and capability of flexible manufacturing.

  11. Six degree of freedom sensor

    DOE Patents [OSTI]

    Vann, C.S.

    1999-03-16

    This small, non-contact optical sensor increases the capability and flexibility of computer controlled machines by detecting its relative position to a workpiece in all six degrees of freedom (DOF). At a fraction of the cost, it is over 200 times faster and up to 25 times more accurate than competing 3-DOF sensors. Applications range from flexible manufacturing to a 6-DOF mouse for computers. Until now, highly agile and accurate machines have been limited by their inability to adjust to changes in their tasks. By enabling them to sense all six degrees of position, these machines can now adapt to new and complicated tasks without human intervention or delay--simplifying production, reducing costs, and enhancing the value and capability of flexible manufacturing. 3 figs.

  12. An integrated distributed processing interface for supercomputers and workstations

    SciTech Connect (OSTI)

    Campbell, J.; McGavran, L.

    1989-01-01

    Access to documentation, communication between multiple processes running on heterogeneous computers, and animation of simulations of engineering problems are typically weak in most supercomputer environments. This presentation will describe how we are improving this situation in the Computer Research and Applications group at Los Alamos National Laboratory. We have developed a tool using UNIX filters and a SunView interface that allows users simple access to documentation via mouse driven menus. We have also developed a distributed application that integrated a two point boundary value problem on one of our Cray Supercomputers. It is controlled and displayed graphically by a window interface running on a workstation screen. Our motivation for this research has been to improve the usual typewriter/static interface using language independent controls to show capabilities of the workstation/supercomputer combination. 8 refs.

  13. Development of Oral Fomulation of SCV-07 for Use in Tuberculosis

    SciTech Connect (OSTI)

    2007-11-16

    An evaluation of the immunomodulatory peptide SCV-07 was conducted as a possible therapeutic treatment for tuberculosis. This evaluation included mouse models, clinical trials and various forms of the drug such as liquid injection and development of an oral pill. It was found that SCV-07 significantly increased the survival rate of animals infected with lethal doses of Mycobacterium bovis. It enhanced the functional activity of macrophages in a dose-dependent fashion. The combination of SCV-07 with bacteriostatic drugs, such as izoniazid, was particularly effective. Phase II clinical trials in a TB clinic demonstrated that the usage of the injection form of SCV-07 for lung TB treatment in combination with standard chemotherapy decreased the quantity of patients with positive sputum assays for Mycobacteria, promoted healing of cavities in lungs, stabilized parameters of cell immunity, and resulted in a significant improvement in the general condition of patients. Clinical trials results of the oral drug form are still being evaluated.

  14. Apolipoprotein gene involved in lipid metabolism

    DOE Patents [OSTI]

    Rubin, Edward; Pennacchio, Len A.

    2007-07-03

    Methods and materials for studying the effects of a newly identified human gene, APOAV, and the corresponding mouse gene apoAV. The sequences of the genes are given, and transgenic animals which either contain the gene or have the endogenous gene knocked out are described. In addition, single nucleotide polymorphisms (SNPs) in the gene are described and characterized. It is demonstrated that certain SNPs are associated with diseases involving lipids and triglycerides and other metabolic diseases. These SNPs may be used alone or with SNPs from other genes to study individual risk factors. Methods for intervention in lipid diseases, including the screening of drugs to treat lipid-related or diabetic diseases are also disclosed.

  15. Reconstructing cerebrovascular networks under local physiological constraints by integer programming

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Rempfler, Markus; Schneider, Matthias; Ielacqua, Giovanna D.; Xiao, Xianghui; Stock, Stuart R.; Klohs, Jan; Szekely, Gabor; Andres, Bjoern; Menze, Bjoern H.

    2015-04-23

    We introduce a probabilistic approach to vessel network extraction that enforces physiological constraints on the vessel structure. The method accounts for both image evidence and geometric relationships between vessels by solving an integer program, which is shown to yield the maximum a posteriori (MAP) estimate to the probabilistic model. Starting from an over-connected network, it is pruning vessel stumps and spurious connections by evaluating the local geometry and the global connectivity of the graph. We utilize a high-resolution micro computed tomography (µCT) dataset of a cerebrovascular corrosion cast to obtain a reference network and learn the prior distributions of ourmore » probabilistic model. As a result, we perform experiments on micro magnetic resonance angiography (µMRA) images of mouse brains and discuss properties of the networks obtained under different tracking and pruning approaches.« less

  16. Rapid and noncontact photoacoustic tomography imaging system using an interferometer with high-speed phase modulation technique

    SciTech Connect (OSTI)

    Liu, Jun; Tang, Zhilie; Wu, Yongbo; Wang, Yi

    2015-04-15

    We designed, fabricated, and tested a rapid and noncontact photoacoustic tomography (PAT) imaging system using a low-coherence interferometer with high-speed phase modulation technique. Such a rapid and noncontact probing system can greatly decrease the time of imaging. The proposed PAT imaging system is experimentally verified by capturing images of a simulated tissue sample and the blood vessels within the ear flap of a mouse (pinna) in vivo. The axial and lateral resolutions of the system are evaluated at 45 and ∼15 μm, respectively. The imaging depth of the system is 1 mm in a special phantom. Our results show that the proposed system opens a promising way to realize noncontact, real-time PAT.

  17. Systematic identification of genes and transduction pathways involved in radio-adaptive response

    SciTech Connect (OSTI)

    Wu, Honglu

    2015-05-22

    Low doses of radiation have been shown to protect against the biological effects of later exposure to toxic levels of radiation. In this study, we propose to identify the molecular mechanisms of this adaptive response by systematically identifying the genes that play a role in radio-protection. In the original proposal, a human cell line that is well-documented to exhibit the radio-adaptive effect was to be used. In this revised study plan, we will use a mouse model, C57BL/6, which has also been well investigated for radio-adaptation. The goal of the proposed study is to enhance our understanding of cellular responses to low doses of radiation exposure at the molecular level.

  18. Protein A suppresses immune responses during Staphylococcus aureus bloodstream infection in guinea pigs

    SciTech Connect (OSTI)

    Kim, Hwan Keun; Falugi, Fabiana; Thomer, Lena; Missiakas, Dominique M.; Schneewind, Olaf

    2015-01-06

    Staphylococcus aureus infection is not associated with the development of protective immunity, and disease relapses occur frequently. We hypothesize that protein A, a factor that binds immunoglobulin Fcγ and cross-links VH3 clan B cell receptors (IgM), is the staphylococcal determinant for host immune suppression. To test this, vertebrate IgM was examined for protein A cross-linking. High VH3 binding activity occurred with human and guinea immunoglobulin, whereas mouse and rabbit immunoglobulins displayed little and no binding, respectively. Establishing a guinea pig model of S. aureus bloodstream infection, we show that protein A functions as a virulence determinant and suppresses host B cell responses. Immunization with SpAKKAA, which cannot bind immunoglobulin, elicits neutralizing antibodies that enable guinea pigs to develop protective immunity.

  19. Structural Genomics of Protein Phosphatases

    SciTech Connect (OSTI)

    Almo,S.; Bonanno, J.; Sauder, J.; Emtage, S.; Dilorenzo, T.; Malashkevich, V.; Wasserman, S.; Swaminathan, S.; Eswaramoorthy, S.; et al

    2007-01-01

    The New York SGX Research Center for Structural Genomics (NYSGXRC) of the NIGMS Protein Structure Initiative (PSI) has applied its high-throughput X-ray crystallographic structure determination platform to systematic studies of all human protein phosphatases and protein phosphatases from biomedically-relevant pathogens. To date, the NYSGXRC has determined structures of 21 distinct protein phosphatases: 14 from human, 2 from mouse, 2 from the pathogen Toxoplasma gondii, 1 from Trypanosoma brucei, the parasite responsible for African sleeping sickness, and 2 from the principal mosquito vector of malaria in Africa, Anopheles gambiae. These structures provide insights into both normal and pathophysiologic processes, including transcriptional regulation, regulation of major signaling pathways, neural development, and type 1 diabetes. In conjunction with the contributions of other international structural genomics consortia, these efforts promise to provide an unprecedented database and materials repository for structure-guided experimental and computational discovery of inhibitors for all classes of protein phosphatases.

  20. Enhanced-locality fiber-optic two-photon-fluorescence live-brain interrogation

    SciTech Connect (OSTI)

    Fedotov, I. V.; Doronina-Amitonova, L. V.; Sidorov-Biryukov, D. A.; Fedotov, A. B.; Anokhin, K. V.; Kilin, S. Ya.; Sakoda, K.; Zheltikov, A. M.

    2014-02-24

    Two-photon excitation is shown to substantially enhance the locality of fiber-based optical interrogation of strongly scattering biotissues. In our experiments, a high-numerical-aperture, large-core-are fiber probe is used to deliver the 200-fs output of a 100-MHz mode-locked ytterbium fiber laser to samples of live mouse brain, induce two-photon fluorescence of nitrogen–vacancy centers in diamond markers in brain sample. Fiber probes with a high numerical aperture and a large core area are shown to enable locality enhancement in fiber-laser–fiber-probe two-photon brain excitation and interrogation without sacrificing the efficiency of fluorescence response collection.

  1. Reconstructing cerebrovascular networks under local physiological constraints by integer programming

    SciTech Connect (OSTI)

    Rempfler, Markus; Schneider, Matthias; Ielacqua, Giovanna D.; Xiao, Xianghui; Stock, Stuart R.; Klohs, Jan; Szekely, Gabor; Andres, Bjoern; Menze, Bjoern H.

    2015-04-23

    We introduce a probabilistic approach to vessel network extraction that enforces physiological constraints on the vessel structure. The method accounts for both image evidence and geometric relationships between vessels by solving an integer program, which is shown to yield the maximum a posteriori (MAP) estimate to the probabilistic model. Starting from an over-connected network, it is pruning vessel stumps and spurious connections by evaluating the local geometry and the global connectivity of the graph. We utilize a high-resolution micro computed tomography (µCT) dataset of a cerebrovascular corrosion cast to obtain a reference network and learn the prior distributions of our probabilistic model. As a result, we perform experiments on micro magnetic resonance angiography (µMRA) images of mouse brains and discuss properties of the networks obtained under different tracking and pruning approaches.

  2. Ensemble Data Analysis ENvironment (EDEN)

    Energy Science and Technology Software Center (OSTI)

    2012-08-01

    The EDEN toolkit facilitates exploratory data analysis and visualization of global climate model simulation datasets. EDEN provides an interactive graphical user interface (GUI) that helps the user visually construct dynamic queries of the characteristically large climate datasets using temporal ranges, variable selections, and geographic areas of interest. EDEN reads the selected data into a multivariate visualization panel which features an extended implementation of parallel coordinates plots as well as interactive scatterplots. The user can querymore » data in the visualization panel using mouse gestures to analyze different ranges of data. The visualization panel provides coordinated multiple views whereby selections made in one plot are propagated to the other plots.« less

  3. Phase contrast portal imaging using synchrotron radiation

    SciTech Connect (OSTI)

    Umetani, K.; Kondoh, T.

    2014-07-15

    Microbeam radiation therapy is an experimental form of radiation treatment with great potential to improve the treatment of many types of cancer. We applied a synchrotron radiation phase contrast technique to portal imaging to improve targeting accuracy for microbeam radiation therapy in experiments using small animals. An X-ray imaging detector was installed 6.0 m downstream from an object to produce a high-contrast edge enhancement effect in propagation-based phase contrast imaging. Images of a mouse head sample were obtained using therapeutic white synchrotron radiation with a mean beam energy of 130 keV. Compared to conventional portal images, remarkably clear images of bones surrounding the cerebrum were acquired in an air environment for positioning brain lesions with respect to the skull structure without confusion with overlapping surface structures.

  4. Modification of the effects of continuous low dose rate irradiation by concurrent chemotherapy infusion

    SciTech Connect (OSTI)

    Fu, K.K.; Rayner, P.A.; Lam, K.N.

    1984-08-01

    The combined effects of continuous low dose rate irradiation (CLDRI) and concurrent infusion of bleomycin, cyclophosphamide, cis-platinum, 5-fluorouracil, actinomycin D, and mitomycin C were studied in the SCC VII/SF tumor, a squamous cell carcinoma and the jejunal crypt cells in the mouse. For the SCC VII/SF tumor, enhanced cell killing was seen with each of the six drugs when infused concurrently with CLDRI; the greatest enhancement was seen with mitomycin C and cis-platinum. For the jejunal crypt cells, enhanced cell killing was seen primarily with bleomycin. The authors results suggest a therapeutic gain with concurrent CLDRI and chemotherapy infusion for five of the six chemotherapeutic drugs studied with the exception of bleomycin.

  5. Properties of zirconia thin films deposited by laser ablation

    SciTech Connect (OSTI)

    Cancea, V. N.; Filipescu, M.; Colceag, D.; Dinescu, M.; Mustaciosu, C.

    2013-11-13

    Zirconia thin films have been deposited by laser ablation of a ceramic ZrO{sub 2} target in vacuum or in oxygen background at 0.01 mbar. The laser beam generated by an ArF laser (λ=193 nm, ν=40 Hz) has been focalized on the target through a spherical lens at an incident angle of 45°. The laser fluence has been established to a value from 2.0 to 3.4 Jcm{sup −2}. A silicon (100) substrate has been placed parallel to the target, at a distance of 4 cm, and subsequently has been heated to temperatures ranging between 300 °C and 600 °C. Thin films morphology has been characterized by atomic force microscopy and secondary ion mass spectrometry. Biocompatibility of these thin films has been assessed by studying the cell attachment of L929 mouse fibroblasts.

  6. Using the Gene Ontology to Enrich Biological Pathways

    SciTech Connect (OSTI)

    Sanfilippo, Antonio P.; Baddeley, Robert L.; Beagley, Nathaniel; McDermott, Jason E.; Riensche, Roderick M.; Taylor, Ronald C.; Gopalan, Banu

    2009-12-10

    Most current approaches to automatic pathway generation are based on a reverse engineering approach in which pathway plausibility is solely derived from microarray gene expression data. These approaches tend to lack in generality and offer no independent validation as they are too reliant on the pathway observables that guide pathway generation. By contrast, alternative approaches that use prior biological knowledge to validate pathways inferred from gene expression data may err in the opposite direction as the prior knowledge is usually not sufficiently tuned to the pathology of focus. In this paper, we present a novel pathway generation approach that combines insights from the reverse engineering and knowledge-based approaches to increase the biological plausibility of automatically generated regulatory networks and describe an application of this approach to transcriptional data from a mouse model of neuroprotection during stroke.

  7. Protein A suppresses immune responses during Staphylococcus aureus bloodstream infection in guinea pigs

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Kim, Hwan Keun; Falugi, Fabiana; Thomer, Lena; Missiakas, Dominique M.; Schneewind, Olaf

    2015-01-06

    Staphylococcus aureus infection is not associated with the development of protective immunity, and disease relapses occur frequently. We hypothesize that protein A, a factor that binds immunoglobulin Fcγ and cross-links VH3 clan B cell receptors (IgM), is the staphylococcal determinant for host immune suppression. To test this, vertebrate IgM was examined for protein A cross-linking. High VH3 binding activity occurred with human and guinea immunoglobulin, whereas mouse and rabbit immunoglobulins displayed little and no binding, respectively. Establishing a guinea pig model of S. aureus bloodstream infection, we show that protein A functions as a virulence determinant and suppresses host Bmore » cell responses. Immunization with SpAKKAA, which cannot bind immunoglobulin, elicits neutralizing antibodies that enable guinea pigs to develop protective immunity.« less

  8. Multivariate Data EXplorer (MDX)

    Energy Science and Technology Software Center (OSTI)

    2012-08-01

    The MDX toolkit facilitates exploratory data analysis and visualization of multivariate datasets. MDX provides and interactive graphical user interface to load, explore, and modify multivariate datasets stored in tabular forms. MDX uses an extended version of the parallel coordinates plot and scatterplots to represent the data. The user can perform rapid visual queries using mouse gestures in the visualization panels to select rows or columns of interest. The visualization panel provides coordinated multiple views wherebymore » selections made in one plot are propagated to the other plots. Users can also export selected data or reconfigure the visualization panel to explore relationships between columns and rows in the data.« less

  9. DEVELOPMENT OF AN INSPECTION PLATFORM AND A SUITE OF SENSORS FOR ASSESSING CORROSION AND MECHANICAL DAMAGE ON UNPIGGABLE TRANSMISSION MAINS

    SciTech Connect (OSTI)

    George C. Vradis; William Leary

    2004-03-01

    The National Energy Technology Laboratory of the US Department of Energy (under Award DE-FC26-02NT41645) and the NYSEARCH Committee of the Northeast Gas Association (previous the New York Gas Group), have sponsored research to develop a robotic pipeline inspection system capable of navigation through the typical physical and operational obstacles that make transmission and distribution pipelines unpiggable. The research contractors, Foster-Miller and GE Oil & Gas (PII North America) have performed an engineering study and developed a conceptual design that meets all the requirements for navigating and inspecting unpiggable transmission pipelines. Based on Foster-Miller's previous efforts developing the Pipe Mouse robot, the RoboScan inspection robot (Figure ES-1) meets the navigational and physical challenges of unpiggable pipelines through an innovative modular platform design, segmented MFL inspection modules and improvements to the inter-module coupling design.

  10. Applications in Data-Intensive Computing

    SciTech Connect (OSTI)

    Shah, Anuj R.; Adkins, Joshua N.; Baxter, Douglas J.; Cannon, William R.; Chavarra-Miranda, Daniel; Choudhury, Sutanay; Gorton, Ian; Gracio, Deborah K.; Halter, Todd D.; Jaitly, Navdeep; Johnson, John R.; Kouzes, Richard T.; Macduff, Matt C.; Marquez, Andres; Monroe, Matthew E.; Oehmen, Christopher S.; Pike, William A.; Scherrer, Chad; Villa, Oreste; Webb-Robertson, Bobbie-Jo M.; Whitney, Paul D.; Zuljevic, Nino

    2010-04-01

    This book chapter, to be published in Advances in Computers, Volume 78, in 2010 describes applications of data intensive computing (DIC). This is an invited chapter resulting from a previous publication on DIC. This work summarizes efforts coming out of the PNNL's Data Intensive Computing Initiative. Advances in technology have empowered individuals with the ability to generate digital content with mouse clicks and voice commands. Digital pictures, emails, text messages, home videos, audio, and webpages are common examples of digital content that are generated on a regular basis. Data intensive computing facilitates human understanding of complex problems. Data-intensive applications provide timely and meaningful analytical results in response to exponentially growing data complexity and associated analysis requirements through the development of new classes of software, algorithms, and hardware.

  11. Institute for Molecular Medicine Research Program

    SciTech Connect (OSTI)

    Phelps, Michael E

    2012-12-14

    The objectives of the project are the development of new Positron Emission Tomography (PET) imaging instrumentation, chemistry technology platforms and new molecular imaging probes to examine the transformations from normal cellular and biological processes to those of disease in pre-clinical animal models. These technology platforms and imaging probes provide the means to: 1. Study the biology of disease using pre-clinical mouse models and cells. 2. Develop molecular imaging probes for imaging assays of proteins in pre-clinical models. 3. Develop imaging assays in pre-clinical models to provide to other scientists the means to guide and improve the processes for discovering new drugs. 4. Develop imaging assays in pre-clinical models for others to use in judging the impact of drugs on the biology of disease.

  12. Palmitoylation of SARS-CoV S protein is necessary for partitioning into detergent-resistant membranes and cell-cell fusion but not interaction with M protein

    SciTech Connect (OSTI)

    McBride, Corrin E.; Machamer, Carolyn E.

    2010-09-15

    Coronaviruses are enveloped RNA viruses that generally cause mild disease in humans. However, the recently emerged coronavirus that caused severe acute respiratory syndrome (SARS-CoV) is the most pathogenic human coronavirus discovered to date. The SARS-CoV spike (S) protein mediates virus entry by binding cellular receptors and inducing fusion between the viral envelope and the host cell membrane. Coronavirus S proteins are palmitoylated, which may affect function. Here, we created a non-palmitoylated SARS-CoV S protein by mutating all nine cytoplasmic cysteine residues. Palmitoylation of SARS-CoV S was required for partitioning into detergent-resistant membranes and for cell-cell fusion. Surprisingly, however, palmitoylation of S was not required for interaction with SARS-CoV M protein. This contrasts with the requirement for palmitoylation of mouse hepatitis virus S protein for interaction with M protein and may point to important differences in assembly and infectivity of these two coronaviruses.

  13. Structural and Functional Profiling of the Human Histone Methyltransferase SMYD3

    SciTech Connect (OSTI)

    Foreman, Kenneth W.; Brown, Mark; Park, Frances; Emtage, Spencer; Harriss, June; Das, Chhaya; Zhu, Li; Crew, Andy; Arnold, Lee; Shaaban, Salam; Tucker, Philip

    2012-10-23

    The SET and MYND Domain (SMYD) proteins comprise a unique family of multi-domain SET histone methyltransferases that are implicated in human cancer progression. Here we report an analysis of the crystal structure of the full length human SMYD3 in a complex with an analog of the S-adenosyl methionine (SAM) methyl donor cofactor. The structure revealed an overall compact architecture in which the 'split-SET' domain adopts a canonical SET domain fold and closely assembles with a Zn-binding MYND domain and a C-terminal superhelical 9 ?-helical bundle similar to that observed for the mouse SMYD1 structure. Together, these structurally interlocked domains impose a highly confined binding pocket for histone substrates, suggesting a regulated mechanism for its enzymatic activity. Our mutational and biochemical analyses confirm regulatory roles of the unique structural elements both inside and outside the core SET domain and establish a previously undetected preference for trimethylation of H4K20.

  14. Giant Magnetoresistive Sensors and Magnetic Labels for Chip-Scale Detection of Immunosorbent Assays

    SciTech Connect (OSTI)

    Rachel Lora Millen

    2005-12-17

    The combination of giant magnetoresistive sensors, magnetic labeling strategies, and biomolecule detection is just beginning to be explored. New readout methods and assay formats are necessary for biomolecules detection to flourish. The work presented in this dissertation describes steps toward the creation of a novel detection method for bioassays utilizing giant magnetoresistive sensors as the readout method. The introduction section contains a brief review of some of the current methods of bioassay readout. The theoretical underpinnings of the giant magnetoresistive effect are also discussed. Finally, the more prominent types of giant magnetoresistive sensors are described, as well as their complicated fabrication. Four data chapters follow the introduction; each chapter is presented as a separate manuscript, either already published or soon to be submitted. Chapter 1 presents research efforts toward the production of a bioassay on the surface of a gold-modified GMR sensor. The testing of this methodology involved the capture of goat a-mouse-coated magnetic nanoparticles on the mouse IgG-modified gold surface. The second, third and fourth chapters describe the utilization of a self-referenced sample stick for scanning across the GMR sensor. The sample stick consisted of alternating magnetic reference and bioactive gold addresses. Chapter 2 is concerned with the characterization of both the scanning readout method and the binding and detection of streptavidin-coated magnetic particles to a biotinylated surface. Chapter 3 advances the sample stick readout with the use of the system for detection of a sandwich immunoassay with rabbit IgG proteins. Finally, simultaneous detection of three IgG proteins is demonstrated in Chapter 4. The dissertation is concluded with a brief summary of the research presented and a discussion of the possible future applications and direction of this work.

  15. Efficient myogenic differentiation of human adipose-derived stem cells by the transduction of engineered MyoD protein

    SciTech Connect (OSTI)

    Sung, Min Sun; Biosystems and Bioengineering Program, University of Science and Technology , Daejeon 305-350 ; Mun, Ji-Young; Kwon, Ohsuk; Biosystems and Bioengineering Program, University of Science and Technology , Daejeon 305-350 ; Kwon, Ki-Sun; Oh, Doo-Byoung; Biosystems and Bioengineering Program, University of Science and Technology , Daejeon 305-350

    2013-07-19

    Highlights: •MyoD was engineered to contain protein transduction domain and endosome-disruptive INF7 peptide. •The engineered MyoD-IT showed efficient nuclear targeting through an endosomal escape by INF7 peptide. •By applying MyoD-IT, human adipose-derived stem cells (hASCs) were differentiated into myogenic cells. •hASCs differentiated by applying MyoD-IT fused to myotubes through co-culturing with mouse myoblasts. •Myogenic differentiation using MyoD-IT is a safe method without the concern of altering the genome. -- Abstract: Human adipose-derived stem cells (hASCs) have great potential as cell sources for the treatment of muscle disorders. To provide a safe method for the myogenic differentiation of hASCs, we engineered the MyoD protein, a key transcription factor for myogenesis. The engineered MyoD (MyoD-IT) was designed to contain the TAT protein transduction domain for cell penetration and the membrane-disrupting INF7 peptide, which is an improved version of the HA2 peptide derived from influenza. MyoD-IT showed greatly improved nuclear targeting ability through an efficient endosomal escape induced by the pH-sensitive membrane disruption of the INF7 peptide. By applying MyoD-IT to a culture, hASCs were efficiently differentiated into long spindle-shaped myogenic cells expressing myosin heavy chains. Moreover, these cells differentiated by an application of MyoD-IT fused to myotubes with high efficiency through co-culturing with mouse C2C12 myoblasts. Because internalized proteins can be degraded in cells without altering the genome, the myogenic differentiation of hASCs using MyoD-IT would be a safe and clinically applicable method.

  16. Coordinated Regulation of Virulence during Systemic Infection of Salmonella enterica serovar Typhimurium

    SciTech Connect (OSTI)

    Yoon, Hyunjin; McDermott, Jason E.; Porwollik, Steffen; Mcclelland, Michael; Heffron, Fred

    2009-02-20

    Salmonella must respond to a myriad of environmental cues during infection of a mouse and express specific subsets of genes in a temporal and spatial manner to subvert the host defense mechanisms but these regulatory pathways are poorly established. To unravel how micro-environmental signals are processed and integrated into coordinated action, we constructed in-frame non-polar deletions of 84 regulators inferred to play a role in Salmonella typhimurium virulence and tested them in three virulence assays (intraperitoneal (i.p.), and intragastric (i.g.) infection in BALB/c mice, and persistence in SvJ129 mice). Overall 36 regulators were identified that were less virulent in at least one assay, and of those, 15 regulators were required for systemic mouse infection in an acute infection model. As a first step towards understanding the interplay between a pathogen and its host from a systems biology standpoint we focused on these 15 genes. Transcriptional profiles were obtained for each of these 15 regulators from strains grown under four different environmental conditions. These results as well as publicly available transcriptional profiles were analyzed using both network inference and cluster analysis algorithms. The analysis predicts a regulatory network in which all 15 regulators control a specific set of genes necessary for Salmonella to cause systemic infection. We tested the regulatory model by expressing a subset of the regulators in trans and monitoring transcription of 7 known virulence factors located within Salmonella pathogenicity island 2 (SPI-2). These experiments validated the regulatory model and showed that, for these 7 genes, the response regulator SsrB and the marR type regulator SlyA co-regulate in a regulatory cascade by integrating multiple signals.

  17. The influence of TRP53 in the dose response of radiation-induced apoptosis, DNA repair and genomic stability in murine haematopoietic cells

    SciTech Connect (OSTI)

    Lemon, Jennifer A.; Taylor, Kristina; Verdecchia, Kyle; Phan, Nghi; Boreham, Douglas R.

    2014-01-01

    Apoptotic and DNA damage endpoints are frequently used as surrogate markers of cancer risk, and have been well-studied in the Trp53+/- mouse model. We report the effect of differing Trp53 gene status on the dose response of ionizing radiation exposures (0.01-2 Gy), with the unique perspective of determining if effects of gene status remain at extended time points. Here we report no difference in the dose response for radiation-induced DNA double-strand breaks in bone marrow and genomic instability (MN-RET levels) in peripheral blood, between wild-type (Trp53+/+) and heterozygous (Trp53+/-) mice. The dose response for Trp53+/+ mice showed higher initial levels of radiation-induced lymphocyte apoptosis relative to Trp53+/- between 0 and 1 Gy. Although this trend was observed up to 12 hours post-irradiation, both genotypes ultimately reached the same level of apoptosis at 14 hours, suggesting the importance of late-onset p53-independent apoptotic responses in this mouse model. Expected radiation-induced G1 cell cycle delay was observed in Trp53+/+ but not Trp53+/-. Although p53 has an important role in cancer risk, we have shown its influence on radiation dose response can be temporally variable. This research highlights the importance of caution when using haematopoietic endpoints as surrogates to extrapolate radiation-induced cancer risk estimation.

  18. Pharmacokinetic drivers of toxicity for basic molecules: Strategy to lower pKa results in decreased tissue exposure and toxicity for a small molecule Met inhibitor

    SciTech Connect (OSTI)

    Diaz, Dolores; Ford, Kevin A.; Hartley, Dylan P.; Harstad, Eric B.; Cain, Gary R.; Achilles-Poon, Kirsten; Nguyen, Trung; Peng, Jing; Zheng, Zhong; Merchant, Mark; Sutherlin, Daniel P.; Gaudino, John J.; Kaus, Robert; Lewin-Koh, Sock C.; Choo, Edna F.; Liederer, Bianca M.; Dambach, Donna M.

    2013-01-01

    Several toxicities are clearly driven by free drug concentrations in plasma, such as toxicities related to on-target exaggerated pharmacology or off-target pharmacological activity associated with receptors, enzymes or ion channels. However, there are examples in which organ toxicities appear to correlate better with total drug concentrations in the target tissues, rather than with free drug concentrations in plasma. Here we present a case study in which a small molecule Met inhibitor, GEN-203, with significant liver and bone marrow toxicity in preclinical species was modified with the intention of increasing the safety margin. GEN-203 is a lipophilic weak base as demonstrated by its physicochemical and structural properties: high LogD (distribution coefficient) (4.3) and high measured pKa (7.45) due to the basic amine (N-ethyl-3-fluoro-4-aminopiperidine). The physicochemical properties of GEN-203 were hypothesized to drive the high distribution of this compound to tissues as evidenced by a moderately-high volume of distribution (Vd > 3 l/kg) in mouse and subsequent toxicities of the compound. Specifically, the basicity of GEN-203 was decreased through addition of a second fluorine in the 3-position of the aminopiperidine to yield GEN-890 (N-ethyl-3,3-difluoro-4-aminopiperidine), which decreased the volume of distribution of the compound in mouse (Vd = 1.0 l/kg), decreased its tissue drug concentrations and led to decreased toxicity in mice. This strategy suggests that when toxicity is driven by tissue drug concentrations, optimization of the physicochemical parameters that drive tissue distribution can result in decreased drug concentrations in tissues, resulting in lower toxicity and improved safety margins. -- Highlights: ? Lower pKa for a small molecule: reduced tissue drug levels and toxicity. ? New analysis tools to assess electrostatic effects and ionization are presented. ? Chemical and PK drivers of toxicity can be leveraged to improve safety.

  19. Genome Clone Libraries and Data from the Integrated Molecular Analysis of Genomes and their Expression (I.M.A.G.E.) Consortium

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    The I.M.A.G.E. Consortium was initiated in 1993 by four academic groups on a collaborative basis after informal discussions led to a common vision of how to achieve an important goal in the study of the human genome: the Integrated Molecular Analysis of Genomes and their Expression Consortium's primary goal is to create arrayed cDNA libraries and associated bioinformatics tools, and make them publicly available to the research community. The primary organisms of interest include intensively studied mammalian species, including human, mouse, rat and non-human primate species. The Consortium has also focused on several commonly studied model organisms; as part of this effort it has arrayed cDNAs from zebrafish, and Fugu (pufferfish) as well as Xenopus laevis and X. tropicalis (frog). Utilizing high speed robotics, over nine million individual cDNA clones have been arrayed into 384-well microtiter plates, and sufficient replicas have been created to distribute copies both to sequencing centers and to a network of five distributors located worldwide. The I.M.A.G.E. Consortium represents the world's largest public cDNA collection, and works closely with the National Institutes of Health's Mammalian Gene Collection(MGC) to help it achieve its goal of creating a full-length cDNA clone for every human and mouse gene. I.M.A.G.E. is also a member of the ORFeome Collaboration, working to generate a complete set of expression-ready open reading frame clones representing each human gene. Custom informatics tools have been developed in support of these projects to better allow the research community to select clones of interest and track and collect all data deposited into public databases about those clones and their related sequences. I.M.A.G.E. clones are publicly available, free of any royalties, and may be used by anyone agreeing with the Consortium's guidelines.

  20. Studies of acute and chronic radiation injury at the Biological and Medical Research Division, Argonne National Laboratory, 1953-1970: Description of individual studies, data files, codes, and summaries of significant findings

    SciTech Connect (OSTI)

    Grahn, D.; Fox, C.; Wright, B.J.; Carnes, B.A.

    1994-05-01

    Between 1953 and 1970, studies on the long-term effects of external x-ray and {gamma} irradiation on inbred and hybrid mouse stocks were carried out at the Biological and Medical Research Division, Argonne National Laboratory. The results of these studies, plus the mating, litter, and pre-experimental stock records, were routinely coded on IBM cards for statistical analysis and record maintenance. Also retained were the survival data from studies performed in the period 1943-1953 at the National Cancer Institute, National Institutes of Health, Bethesda, Maryland. The card-image data files have been corrected where necessary and refiled on hard disks for long-term storage and ease of accessibility. In this report, the individual studies and data files are described, and pertinent factors regarding caging, husbandry, radiation procedures, choice of animals, and other logistical details are summarized. Some of the findings are also presented. Descriptions of the different mouse stocks and hybrids are included in an appendix; more than three dozen stocks were involved in these studies. Two other appendices detail the data files in their original card-image format and the numerical codes used to describe the animal`s exit from an experiment and, for some studies, any associated pathologic findings. Tabular summaries of sample sizes, dose levels, and other variables are also given to assist investigators in their selection of data for analysis. The archive is open to any investigator with legitimate interests and a willingness to collaborate and acknowledge the source of the data and to recognize appropriate conditions or caveats.

  1. Differential modulation of dibenzo[def,p]chrysene transplacental carcinogenesis: Maternal diets rich in indole-3-carbinol versus sulforaphane

    SciTech Connect (OSTI)

    Shorey, Lyndsey E.; Madeen, Erin P.; Atwell, Lauren L.; Ho, Emily; Lhr, Christiane V.; Pereira, Clifford B.; Dashwood, Roderick H.; Williams, David E.

    2013-07-01

    Cruciferous vegetable components have been documented to exhibit anticancer properties. Targets of action span multiple mechanisms deregulated during cancer progression, ranging from altered carcinogen metabolism to the restoration of epigenetic machinery. Furthermore, the developing fetus is highly susceptible to changes in nutritional status and to environmental toxicants. Thus, we have exploited a mouse model of transplacental carcinogenesis to assess the impact of maternal dietary supplementation on cancer risk in offspring. In this study, transplacental and lactational exposure to a maternal dose of 15 mg/Kg B.W. of dibenzo[def,p]chrysene (DBC) resulted in significant morbidity of offspring due to an aggressive T-cell lymphoblastic lymphoma. As in previous studies, indole-3-carbinol (I3C, feed to the dam at 100, 500 or 1000 ppm), derived from cruciferous vegetables, dose-dependently reduced lung tumor multiplicity and also increased offspring survival. Brussels sprout and broccoli sprout powders, selected for their relative abundance of I3C and the bioactive component sulforaphane (SFN), respectively, surprisingly enhanced DBC-induced morbidity and tumorigenesis when incorporated into the maternal diet at 10% wt/wt. Purified SFN, incorporated in the maternal diet at 400 ppm, also decreased the latency of DBC-dependent morbidity. Interestingly, I3C abrogated the effect of SFN when the two purified compounds were administered in equimolar combination (500 ppm I3C and 600 ppm SFN). SFN metabolites measured in the plasma of neonates positively correlated with exposure levels via the maternal diet but not with offspring mortality. These findings provide justification for further study of the safety and bioactivity of cruciferous vegetable phytochemicals at supplemental concentrations during the perinatal period. - Highlights: Dietary supplementation may modulate cancer risk in a mouse model of lymphoma. Cruciferous vegetables may not contain sufficient I3C for transplacental protection. SFN is abundant in cruciferous vegetables and may enhance risk in this model. SFN and its mercapturic acid metabolites were measurable in neonatal plasma.

  2. Toxicogenomic outcomes predictive of forestomach carcinogenesis following exposure to benzo(a)pyrene: Relevance to human cancer risk

    SciTech Connect (OSTI)

    Labib, Sarah Guo, Charles H. Williams, Andrew Yauk, Carole L. White, Paul A. Halappanavar, Sabina

    2013-12-01

    Forestomach tumors are observed in mice exposed to environmental carcinogens. However, the relevance of this data to humans is controversial because humans lack a forestomach. We hypothesize that an understanding of early molecular changes after exposure to a carcinogen in the forestomach will provide mode-of-action information to evaluate the applicability of forestomach cancers to human cancer risk assessment. In the present study we exposed mice to benzo(a)pyrene (BaP), an environmental carcinogen commonly associated with tumors of the rodent forestomach. Toxicogenomic tools were used to profile gene expression response in the forestomach. Adult MutaMouse males were orally exposed to 25, 50, and 75 mg BaP/kg-body-weight/day for 28 consecutive days. The forestomach was collected three days post-exposure. DNA microarrays, real-time RT-qPCR arrays, and protein analyses were employed to characterize responses in the forestomach. Microarray results showed altered expression of 414 genes across all treatment groups ( 1.5 fold; false discovery rate adjusted P ? 0.05). Significant downregulation of genes associated with phase II xenobiotic metabolism and increased expression of genes implicated in antigen processing and presentation, immune response, chemotaxis, and keratinocyte differentiation were observed in treated groups in a dose-dependent manner. A systematic comparison of the differentially expressed genes in the forestomach from the present study to differentially expressed genes identified in human diseases including human gastrointestinal tract cancers using the NextBio Human Disease Atlas showed significant commonalities between the two models. Our results provide molecular evidence supporting the use of the mouse forestomach model to evaluate chemically-induced gastrointestinal carcinogenesis in humans. - Highlights: Benzo(a)pyrene-mediated transcriptomic response in the forestomach was examined. The immunoproteosome subunits and MHC class I pathway were the most affected. Keratinocyte differentiation associated gene expression changes were dose-dependent. Molecular similarities exist between cancers of the forestomach and human stomach.

  3. Cytotoxicity of synthetic cannabinoids on primary neuronal cells of the forebrain: the involvement of cannabinoid CB{sub 1} receptors and apoptotic cell death

    SciTech Connect (OSTI)

    Tomiyama, Ken-ichi; Funada, Masahiko

    2014-01-01

    The abuse of herbal products containing synthetic cannabinoids has become an issue of public concern. The purpose of this paper was to evaluate the acute cytotoxicity of synthetic cannabinoids on mouse brain neuronal cells. Cytotoxicity induced by synthetic cannabinoid (CP-55,940, CP-47,497, CP-47,497-C8, HU-210, JWH-018, JWH-210, AM-2201, and MAM-2201) was examined using forebrain neuronal cultures. These synthetic cannabinoids induced cytotoxicity in the forebrain cultures in a concentration-dependent manner. The cytotoxicity was suppressed by preincubation with the selective CB{sub 1} receptor antagonist AM251, but not with the selective CB{sub 2} receptor antagonist AM630. Furthermore, annexin-V-positive cells were found among the treated forebrain cells. Synthetic cannabinoid treatment induced the activation of caspase-3, and preincubation with a caspase-3 inhibitor significantly suppressed the cytotoxicity. These synthetic cannabinoids induced apoptosis through a caspase-3-dependent mechanism in the forebrain cultures. Our results indicate that the cytotoxicity of synthetic cannabinoids towards primary neuronal cells is mediated by the CB{sub 1} receptor, but not by the CB{sub 2} receptor, and further suggest that caspase cascades may play an important role in the apoptosis induced by these synthetic cannabinoids. In conclusion, excessive synthetic cannabinoid abuse may present a serious acute health concern due to neuronal damage or deficits in the brain. - Highlights: Synthetic cannabinoids (classical cannabinoids, non-classical cannabinoids, and aminoalkylindole derivatives) induce cytotoxicity in mouse forebrain cultures. Synthetic cannabinoid-induced cytotoxicity towards forebrain cultures is mediated by the CB{sub 1} receptor, but not by the CB{sub 2} receptor, and involves caspase-dependent apoptosis. A high concentration of synthetic cannabinoids may be toxic to neuronal cells that express CB{sub 1} receptors.

  4. Chronic subhepatotoxic exposure to arsenic enhances hepatic injury caused by high fat diet in mice

    SciTech Connect (OSTI)

    Tan, Min; Schmidt, Robin H.; Beier, Juliane I.; Department of Medicine, University of Louisville Health Sciences Center, Louisville, KY 40292 ; Watson, Walter H.; University of Louisville Alcohol Research Center, University of Louisville Health Sciences Center, Louisville, KY 40292 ; Zhong, Hai; University of Louisville Alcohol Research Center, University of Louisville Health Sciences Center, Louisville, KY 40292 ; States, J. Christopher; Arteel, Gavin E.

    2011-12-15

    Arsenic is a ubiquitous contaminant in drinking water. Whereas arsenic can be directly hepatotoxic, the concentrations/doses required are generally higher than present in the US water supply. However, physiological/biochemical changes that are alone pathologically inert can enhance the hepatotoxic response to a subsequent stimulus. Such a '2-hit' paradigm is best exemplified in chronic fatty liver diseases. Here, the hypothesis that low arsenic exposure sensitizes liver to hepatotoxicity in a mouse model of non-alcoholic fatty liver disease was tested. Accordingly, male C57Bl/6J mice were exposed to low fat diet (LFD; 13% calories as fat) or high fat diet (HFD; 42% calories as fat) and tap water or arsenic (4.9 ppm as sodium arsenite) for ten weeks. Biochemical and histologic indices of liver damage were determined. High fat diet ({+-} arsenic) significantly increased body weight gain in mice compared with low-fat controls. HFD significantly increased liver to body weight ratios; this variable was unaffected by arsenic exposure. HFD caused steatohepatitis, as indicated by histological assessment and by increases in plasma ALT and AST. Although arsenic exposure had no effect on indices of liver damage in LFD-fed animals, it significantly increased the liver damage caused by HFD. This effect of arsenic correlated with enhanced inflammation and fibrin extracellular matrix (ECM) deposition. These data indicate that subhepatotoxic arsenic exposure enhances the toxicity of HFD. These results also suggest that arsenic exposure might be a risk factor for the development of fatty liver disease in human populations. -- Highlights: Black-Right-Pointing-Pointer Characterizes a mouse model of arsenic enhanced NAFLD. Black-Right-Pointing-Pointer Arsenic synergistically enhances experimental fatty liver disease at concentrations that cause no overt hepatotoxicity alone. Black-Right-Pointing-Pointer This effect is associated with increased inflammation.

  5. SU-E-T-501: Normal Tissue Toxicities of Pulsed Low Dose Rate Radiotherapy and Conventional Radiotherapy: An in Vivo Total Body Irradiation Study

    SciTech Connect (OSTI)

    Cvetkovic, D; Zhang, P; Wang, B; Chen, L; Ma, C

    2014-06-01

    Purpose: Pulsed low dose rate radiotherapy (PLDR) is a re-irradiation technique for therapy of recurrent cancers. We have previously shown a significant difference in the weight and survival time between the mice treated with conventional radiotherapy (CRT) and PLDR using total body irradiation (TBI). The purpose of this study was to investigate the in vivo effects of PLDR on normal mouse tissues.Materials and Methods: Twenty two male BALB/c nude mice, 4 months of age, were randomly assigned into a PLDR group (n=10), a CRT group (n=10), and a non-irradiated control group (n=2). The Siemens Artiste accelerator with 6 MV photon beams was used. The mice received a total of 18Gy in 3 fractions with a 20day interval. The CRT group received the 6Gy dose continuously at a dose rate of 300 MU/min. The PLDR group was irradiated with 0.2Gyx20 pulses with a 3min interval between the pulses. The mice were weighed thrice weekly and sacrificed 2 weeks after the last treatment. Brain, heart, lung, liver, spleen, gastrointestinal, urinary and reproductive organs, and sternal bone marrow were removed, formalin-fixed, paraffin-embedded and stained with H and E. Morphological changes were observed under a microscope. Results: Histopathological examination revealed atrophy in several irradiated organs. The degree of atrophy was mild to moderate in the PLDR group, but severe in the CRT group. The most pronounced morphological abnormalities were in the immune and hematopoietic systems, namely spleen and bone marrow. Brain hemorrhage was seen in the CRT group, but not in the PLDR group. Conclusions: Our results showed that PLDR induced less toxicity in the normal mouse tissues than conventional radiotherapy for the same dose and regimen. Considering that PLDR produces equivalent tumor control as conventional radiotherapy, it would be a good modality for treatment of recurrent cancers.

  6. Mass Spectrometry Imaging of Biological Tissue: An Approach for Multicenter Studies

    SciTech Connect (OSTI)

    Rompp, Andreas; Both, Jean-Pierre; Brunelle, Alain; Heeren, Ronald M.; Laprevote, Olivier; Prideaux, Brendan; Seyer, Alexandre; Spengler, Bernhard; Stoeckli, Markus; Smith, Donald F.

    2015-03-01

    Mass spectrometry imaging has become a popular tool for probing the chemical complexity of biological surfaces. This led to the development of a wide range of instrumentation and preparation protocols. It is thus desirable to evaluate and compare the data output from different methodologies and mass spectrometers. Here, we present an approach for the comparison of mass spectrometry imaging data from different laboratories (often referred to as multicenter studies). This is exemplified by the analysis of mouse brain sections in five laboratories in Europe and the USA. The instrumentation includes matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF), MALDI-QTOF, MALDIFourier transform ion cyclotron resonance (FTICR), atmospheric-pressure (AP)-MALDI-Orbitrap, and cluster TOF-secondary ion mass spectrometry (SIMS). Experimental parameters such as measurement speed, imaging bin width, and mass spectrometric parameters are discussed. All datasets were converted to the standard data format imzML and displayed in a common open-source software with identical parameters for visualization, which facilitates direct comparison of MS images. The imzML conversion also allowed exchange of fully functional MS imaging datasets between the different laboratories. The experiments ranged from overview measurements of the full mouse brain to detailed analysis of smaller features (depending on spatial resolution settings), but common histological features such as the corpus callosum were visible in all measurements. High spatial resolution measurements of AP-MALDI-Orbitrap and TOF-SIMS showed comparable structures in the low-micrometer range. We discuss general considerations for planning and performing multicenter studies in mass spectrometry imaging. This includes details on the selection, distribution, and preparation of tissue samples as well as on data handling. Such multicenter studies in combination with ongoing activities for reporting guidelines, a common data format (imzML) and a public data repository can contribute to more reliability and transparency of MS imaging studies.

  7. Activation of nuclear receptor NR5A2 increases Glut4 expression and glucose metabolism in muscle cells

    SciTech Connect (OSTI)

    Bolado-Carrancio, A.; Riancho, J.A.; Sainz, J.; Rodrguez-Rey, J.C.

    2014-04-04

    Highlights: NR5A2 expression in C2C12 is associated with myotube differentiation. DLPC induces an increase in GLUT4 levels and glucose uptake in C2C12 myotubes. In high glucose conditions the activation of NR5A2 inhibits fatty acids oxidation. - Abstract: NR5A2 is a nuclear receptor which regulates the expression of genes involved in cholesterol metabolism, pluripotency maintenance and cell differentiation. It has been recently shown that DLPC, a NR5A2 ligand, prevents liver steatosis and improves insulin sensitivity in mouse models of insulin resistance, an effect that has been associated with changes in glucose and fatty acids metabolism in liver. Because skeletal muscle is a major tissue in clearing glucose from blood, we studied the effect of the activation of NR5A2 on muscle metabolism by using cultures of C2C12, a mouse-derived cell line widely used as a model of skeletal muscle. Treatment of C2C12 with DLPC resulted in increased levels of expression of GLUT4 and also of several genes related to glycolysis and glycogen metabolism. These changes were accompanied by an increased glucose uptake. In addition, the activation of NR5A2 produced a reduction in the oxidation of fatty acids, an effect which disappeared in low-glucose conditions. Our results suggest that NR5A2, mostly by enhancing glucose uptake, switches muscle cells into a state of glucose preference. The increased use of glucose by muscle might constitute another mechanism by which NR5A2 improves blood glucose levels and restores insulin sensitivity.

  8. IGF-I enhances cellular senescence via the reactive oxygen species-p53 pathway

    SciTech Connect (OSTI)

    Handayaningsih, Anastasia-Evi; Takahashi, Michiko; Fukuoka, Hidenori; Iguchi, Genzo; Nishizawa, Hitoshi; Yamamoto, Masaaki; Suda, Kentaro; Takahashi, Yutaka

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer Cellular senescence plays an important role in tumorigenesis and aging process. Black-Right-Pointing-Pointer We demonstrated IGF-I enhanced cellular senescence in primary confluent cells. Black-Right-Pointing-Pointer IGF-I enhanced cellular senescence in the ROS and p53-dependent manner. Black-Right-Pointing-Pointer These results may explain the underlying mechanisms of IGF-I involvement in tumorigenesis and in regulation of aging. -- Abstract: Cellular senescence is characterized by growth arrest, enlarged and flattened cell morphology, the expression of senescence-associated {beta}-galactosidase (SA-{beta}-gal), and by activation of tumor suppressor networks. Insulin-like growth factor-I (IGF-I) plays a critical role in cellular growth, proliferation, tumorigenesis, and regulation of aging. In the present study, we show that IGF-I enhances cellular senescence in mouse, rat, and human primary cells in the confluent state. IGF-I induced expression of a DNA damage marker, {gamma}H2AX, the increased levels of p53 and p21 proteins, and activated SA-{beta}-gal. In the confluent state, an altered downstream signaling of IGF-I receptor was observed. Treatment with a reactive oxygen species (ROS) scavenger, N-acetylcystein (NAC) significantly suppressed induction of these markers, indicating that ROS are involved in the induction of cellular senescence by IGF-I. In p53-null mouse embryonic fibroblasts, the IGF-I-induced augmentation of SA-{beta}-gal and p21 was inhibited, demonstrating that p53 is required for cellular senescence induced by IGF-I. Thus, these data reveal a novel pathway whereby IGF-I enhances cellular senescence in the ROS and p53-dependent manner and may explain the underlying mechanisms of IGF-I involvement in tumorigenesis and in regulation of aging.

  9. Curcumin blocks interleukin (IL)-2 signaling in T-lymphocytes by inhibiting IL-2 synthesis, CD25 expression, and IL-2 receptor signaling

    SciTech Connect (OSTI)

    Forward, Nicholas A.; Conrad, David M.; Power Coombs, Melanie R.; Doucette, Carolyn D.; Furlong, Suzanne J.; Lin, Tong-Jun; Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia ; Hoskin, David W.

    2011-04-22

    Highlights: {yields} Curcumin inhibits CD4{sup +} T-lymphocyte proliferation. {yields} Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4{sup +} T-lymphocytes. {yields} Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. {yields} IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4{sup +} T-lymphocytes, as well as interleukin-2 (IL-2) and CD25 ({alpha} chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca{sup 2+} release to inhibit I{kappa}B phosphorylation, which is required for nuclear translocation of the transcription factor NF{kappa}B. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4{sup +}CD25{sup +} regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  10. High-throughput behavioral phenotyping of drug and alcohol susceptibility traits in the expanded panel of BXD recombinant inbred strains

    SciTech Connect (OSTI)

    Philip, Vivek M [ORNL; Ansah, T [University of Tennessee Health Science Center, Memphis; Blaha, C, [University of Tennessee Health Science Center, Memphis; Cook, Melloni N. [University of Memphis; Hamre, Kristin M. [University of Tennessee Health Science Center, Memphis; Lariviere, William R [University of Pittsburgh; Matthews, Douglas B [Baylor University; Goldowitz, Daniel [University of British Columbia, Vancouver; Chesler, Elissa J [ORNL

    2010-01-01

    Genetic reference populations, particularly the BXD recombinant inbred strains, are a valuable resource for the discovery of the bio-molecular substrates and genetic drivers responsible for trait variation and co- ariation. This approach can be profitably applied in the analysis of susceptibility and mechanisms of drug and alcohol use disorders for which many predisposing behaviors may predict occurrence and manifestation of increased preference for these substances. Many of these traits are modeled by common mouse behavioral assays, facilitating the detection of patterns and sources of genetic co-regulation of predisposing phenotypes and substance consumption. Members of the Tennessee Mouse Genome Consortium have obtained behavioral phenotype data from 260 measures related to multiple behavioral assays across several domains: self-administration, response to, and withdrawal from cocaine, MDMA, morphine and alcohol; novelty seeking; behavioral despair and related neurological phenomena; pain sensitivity; stress sensitivity; anxiety; hyperactivity; and sleep/wake cycles. All traits have been measured in both sexes and the recently expanded panel of 69 additional BXD recombinant inbred strains (N=69). Sex differences and heritability estimates were obtained for each trait, and a comparison of early (N = 32) and recent BXD RI lines was performed. Primary data is publicly available for heritability, sex difference and genetic analyses using www.GeneNetwork.org. These analyses include QTL detection and genetic analysis of gene expression. Stored results from these analyses are available at http://ontologicaldiscovery.org for comparison to other genomic analysis results. Together with the results of related studies, these data form a public resource for integrative systems genetic analysis of neurobehavioral traits.

  11. Anticancer effect of genistein on BG-1 ovarian cancer growth induced by 17 ?-estradiol or bisphenol A via the suppression of the crosstalk between estrogen receptor alpha and insulin-like growth factor-1 receptor signaling pathways

    SciTech Connect (OSTI)

    Hwang, Kyung-A; Park, Min-Ah; Kang, Nam-Hee; Yi, Bo-Rim; Hyun, Sang-Hwan; Jeung, Eui-Bae; Choi, Kyung-Chul

    2013-11-01

    The interaction between estrogen receptor (ER) and insulin-like growth factor-1 receptor (IGF-1R) signaling pathway plays an important role in proliferation of and resistance to endocrine therapy to estrogen dependent cancers. Estrogen (E2) upregulates the expression of components of IGF-1 system and induces the downstream of mitogenic signaling cascades via phosphorylation of insulin receptor substrate-1 (IRS-1). In the present study, we evaluated the xenoestrogenic effect of bisphenol A (BPA) and antiproliferative activity of genistein (GEN) in accordance with the influence on this crosstalk. BPA was determined to affect this crosstalk by upregulating mRNA expressions of ER? and IGF-1R and inducing phosphorylation of IRS-1 and Akt in protein level in BG-1 ovarian cancer cells as E2 did. In the mouse model xenografted with BG-1 cells, BPA significantly increased a tumor burden of mice and expressions of ER?, pIRS-1, and cyclin D1 in tumor mass compared to vehicle, indicating that BPA induces ovarian cancer growth by promoting the crosstalk between ER and IGF-1R signals. On the other hand, GEN effectively reversed estrogenicity of BPA by reversing mRNA and protein expressions of ER?, IGF-1R, pIRS-1, and pAkt induced by BPA in cellular model and also significantly decreased tumor growth and in vivo expressions of ER?, pIRS-1, and pAkt in xenografted mouse model. Also, GEN was confirmed to have an antiproliferative effect by inducing apoptotic signaling cascades. Taken together, these results suggest that GEN effectively reversed the increased proliferation of BG-1 ovarian cancer by suppressing the crosstalk between ER? and IGF-1R signaling pathways upregulated by BPA or E2.

  12. SU-E-T-20: A Novel Hybrid CBCT, Bioluminescence and Fluorescence Tomography System for Preclinical Radiation Research

    SciTech Connect (OSTI)

    Zhang, B; Eslami, S; Iordachita, I; Yang, Y; Patterson, M; Wong, J; Wang, K

    2014-06-01

    Purpose: A novel standalone bioluminescence and fluorescence tomography (BLT and FT) system equipped with high resolution CBCT has been built in our group. In this work, we present the system calibration method and validate our system in both phantom and in vivo environment. Methods: The CBCT is acquired by rotating the animal stage while keeping the x-ray source and detector panel static. The optical signal is reflected by the 3-mirror system to a multispectral filter set and then delivered to the CCD camera with f/1.4 lens mounted. Nine fibers passing through the stage and in contact with the mouse skin serve as the light sources for diffuse optical tomography (DOT) and FT. The anatomical information and optical properties acquired from the CBCT and DOT, respectively, are used as the priori information to improve the BLT/FT reconstruction accuracy. Flat field correction for the optical system was acquired at multiple wavelengths. A home-built phantom is used to register the optical and CBCT coordinates. An absolute calibration relating the CCD photon counts rate to the light fluence rate emitted at animal surface was developed to quantify the bioluminescence power or fluorophore concentration. Results: An optical inhomogeneous phantom with 2 light sources (3mm separation) imbedded is used to test the system. The optical signal is mapped onto the mesh generated from CBCT for optical reconstruction. Our preliminary results show that the center of mass can be reconstructed within 2.8mm accuracy. A live mouse with the light source imbedded is also used to validate our system. Liver or lung metastatic luminescence tumor model will be used for further testing. Conclusion: This hybrid system transforms preclinical research to a level that even sub-palpable volume of cells can be imaged rapidly and non-invasively, which largely extends the scope of radiobiological research. The research is supported by the NCI grant R01CA158100-01.

  13. Hepatic lipid profiling of deer mice fed ethanol using {sup 1}H and {sup 31}P NMR spectroscopy: A dose-dependent subchronic study

    SciTech Connect (OSTI)

    Fernando, Harshica; Bhopale, Kamlesh K.; Boor, Paul J.; Ansari, G.A. Shakeel; Kaphalia, Bhupendra S.

    2012-11-01

    Chronic alcohol abuse is a 2nd major cause of liver disease resulting in significant morbidity and mortality. Alcoholic liver disease (ALD) is characterized by a wide spectrum of pathologies starting from fat accumulation (steatosis) in early reversible stage to inflammation with or without fibrosis and cirrhosis in later irreversible stages. Previously, we reported significant steatosis in the livers of hepatic alcohol dehydrogenase (ADH)-deficient (ADH{sup −}) vs. hepatic ADH-normal (ADH{sup +}) deer mice fed 4% ethanol daily for 2 months [Bhopale et al., 2006, Alcohol 39, 179–188]. However, ADH{sup −} deer mice fed 4% ethanol also showed a significant mortality. Therefore, a dose-dependent study was conducted to understand the mechanism and identify lipid(s) involved in the development of ethanol-induced fatty liver. ADH{sup −} and ADH{sup +} deer mice fed 1, 2 or 3.5% ethanol daily for 2 months and fatty infiltration in the livers were evaluated by histology and by measuring dry weights of extracted lipids. Lipid metabolomic changes in extracted lipids were determined by proton ({sup 1}H) and {sup 31}phosphorus ({sup 31}P) nuclear magnetic resonance (NMR) spectroscopy. The NMR data was analyzed by hierarchical clustering (HC) and principle component analysis (PCA) for pattern recognition. Extensive vacuolization by histology and significantly increased dry weights of total lipids found only in the livers of ADH{sup −} deer mice fed 3.5% ethanol vs. pair-fed controls suggest a dose-dependent formation of fatty liver in ADH{sup −} deer mouse model. Analysis of NMR data of ADH{sup −} deer mice fed 3.5% ethanol vs. pair-fed controls shows increases for total cholesterol, esterified cholesterol, fatty acid methyl esters (FAMEs), triacylglycerides and unsaturation, and decreases for free cholesterol, phospholipids and allylic and diallylic protons. Certain classes of neutral lipids (cholesterol esters, fatty acyl chain (-COCH{sub 2}-) and FAMEs) were also mildly increased in ADH{sup −} deer mice fed 1 or 2% ethanol. Only small increases were observed for allylic and diallylic protons, FAMEs and unsaturations in ADH{sup +} deer mice fed 3.5% ethanol vs. pair-fed controls. PCA of NMR data showed increased clustering by gradual separation of ethanol-fed ADH{sup −} deer mice groups from their respective pair-fed control groups and corresponding ethanol-fed ADH{sup +} deer mice groups. Our data indicate that dose of ethanol and hepatic ADH deficiency are two key factors involved in initiation and progression of alcoholic fatty liver disease. Further studies on characterization of individual lipid entities and associated metabolic pathways altered in our deer mouse model after different durations of ethanol feeding could be important to delineate mechanism(s) and identify potential biomarker candidate(s) of early stage ALD. -- Highlights: ► Dose-dependent ethanol-induced fatty liver was studied in deer mouse model. ► A NMR-based lipidomic approach with histology and dry lipid weights was used. ► We used principal component analysis (PCA) to analyze the NMR lipidomic data. ► Dose-dependent clustering patterns by PCA were compared among the groups.

  14. LDRD final report on confinement of cluster fusion plasmas with magnetic fields.

    SciTech Connect (OSTI)

    Argo, Jeffrey W.; Kellogg, Jeffrey W.; Headley, Daniel Ignacio; Stoltzfus, Brian Scott; Waugh, Caleb J.; Lewis, Sean M.; Porter, John Larry, Jr.; Wisher, Matthew; Struve, Kenneth William; Savage, Mark Edward; Quevedo, Hernan J.; Bengtson, Roger

    2011-11-01

    Two versions of a current driver for single-turn, single-use 1-cm diameter magnetic field coils have been built and tested at the Sandia National Laboratories for use with cluster fusion experiments at the University of Texas in Austin. These coils are used to provide axial magnetic fields to slow radial loss of electrons from laser-produced deuterium plasmas. Typical peak field strength achievable for the two-capacitor system is 50 T, and 200 T for the ten-capacitor system. Current rise time for both systems is about 1.7 {mu}s, with peak current of 500 kA and 2 MA, respectively. Because the coil must be brought to the laser, the driver needs to be portable and drive currents in vacuum. The drivers are complete but laser-plasma experiments are still in progress. Therefore, in this report, we focus on system design, initial tests, and performance characteristics of the two-capacitor and ten-capacitors systems. The questions of whether a 200 T magnetic field can retard the breakup of a cluster-fusion plasma, and whether this field can enhance neutron production have not yet been answered. However, tools have been developed that will enable producing the magnetic fields needed to answer these questions. These are a two-capacitor, 400-kA system that was delivered to the University of Texas in 2010, and a 2-MA ten-capacitor system delivered this year. The first system allowed initial testing, and the second system will be able to produce the 200 T magnetic fields needed for cluster fusion experiments with a petawatt laser. The prototype 400-kA magnetic field driver system was designed and built to test the design concept for the system, and to verify that a portable driver system could be built that delivers current to a magnetic field coil in vacuum. This system was built copying a design from a fixed-facility, high-field machine at LANL, but made to be portable and to use a Z-machine-like vacuum insulator and vacuum transmission line. This system was sent to the University of Texas in Austin where magnetic fields up to 50 T have been produced in vacuum. Peak charge voltage and current for this system have been 100 kV and 490 kA. It was used this last year to verify injection of deuterium and surrogate clusters into these small, single-turn coils without shorting the coil. Initial test confirmed the need to insulate the inner surface of the coil, which requires that the clusters must be injected through small holes in an insulator. Tests with a low power laser confirmed that it is possible to inject clusters into the magnetic field coils through these holes without destroying the clusters. The university team also learned the necessity of maintaining good vacuum to avoid insulator, transmission line, and coil shorting. A 200-T, 2 MA system was also constructed using the experience from the first design to make the pulsed-power system more robust. This machine is a copy of the prototype design, but with ten 100-kV capacitors versus the two used in the prototype. It has additional inductance in the switch/capacitor unit to avoid breakdown seen in the prototype design. It also has slightly more inductance at the cable connection to the vacuum chamber. With this design we have been able to demonstrate 1 MA current into a 1 cm diameter coil with the vacuum chamber at air pressure. Circuit code simulations, including the additional inductance with the new design, agree well with the measured current at a charge voltage of 40 kV with a short circuit load, and at 50 kV with a coil. The code also predicts that with a charge voltage of 97 kV we will be able to get 2 MA into a 1 cm diameter coil, which will be sufficient for 200 T fields. Smaller diameter or multiple-turn coils will be able to achieve even higher fields, or be able to achieve 200-T fields with lower charge voltage. Work is now proceeding at the university under separate funding to verify operation at the 2-MA level, and to address issues of debris mitigation, measurement of the magnetic field, and operation in vacuum. We anticipate operation at full current with single

  15. Lung autophagic response following exposure of mice to whole body irradiation, with and without amifostine

    SciTech Connect (OSTI)

    Zois, Christos E.; Giatromanolaki, Alexandra; Kainulainen, Heikki; Botaitis, Sotirios; Torvinen, Sira; Simopoulos, Constantinos; Kortsaris, Alexandros; Sivridis, Efthimios; Koukourakis, Michael I.

    2011-01-07

    Research highlights: {yields} We investigated the effect 6 Gy of WBI on the autophagic machinery of normal mouse lung. {yields} Irradiation induces dysfunction of the autophagic machinery in normal lung, characterized by decreased transcription of the LC3A/Beclin-1 mRNA and accumulation of the LC3A, and p62 proteins. {yields} The membrane bound LC3A-II protein levels increased in the cytosolic fraction (not in the pellet), contrasting the patterns noted after starvation-induced autophagy. {yields} Administration of amifostine, reversed all the LC3A and p62 findings, suggesting protection of the normal autophagic function. -- Abstract: Purpose: The effect of ionizing irradiation on the autophagic response of normal tissues is largely unexplored. Abnormal autophagic function may interfere the protein quality control leading to cell degeneration and dysfunction. This study investigates its effect on the autophagic machinery of normal mouse lung. Methods and materials: Mice were exposed to 6 Gy of whole body {gamma}-radiation and sacrificed at various time points. The expression of MAP1LC3A/LC3A/Atg8, beclin-1, p62/sequestosome-1 and of the Bnip3 proteins was analyzed. Results: Following irradiation, the LC3A-I and LC3A-II protein levels increased significantly at 72 h and 7 days. Strikingly, LC3A-II protein was increased (5.6-fold at 7 days; p < 0.001) only in the cytosolic fraction, but remained unchanged in the membrane fraction. The p62 protein, was significantly increased in both supernatant and pellet fraction (p < 0.001), suggesting an autophagosome turnover deregulation. These findings contrast the patterns of starvation-induced autophagy up-regulation. Beclin-1 levels remained unchanged. The Bnip3 protein was significantly increased at 8 h, but it sharply decreased at 72 h (p < 0.05). Administration of amifostine (200 mg/kg), 30 min before irradiation, reversed all the LC3A and p62 findings on blots, suggesting restoration of the normal autophagic function. The LC3A and Beclin1 mRNA levels significantly declined following irradiation (p < 0.01), whereas Bnip3 levels increased. Conclusions: It is suggested that irradiation induces dysfunction of the autophagic machinery in normal lung, characterized by decreased transcription of the LC3A/Beclin-1 mRNA and accumulation of the LC3A, and p62 proteins. Whether this is due to defective maturation or to aberrant degradation of the autophagosomes requires further investigation.

  16. Lamellar crystalline self-assembly behaviour and solid lipid nanoparticles of a palmityl prodrug analogue of Capecitabine?A chemotherapy agent

    SciTech Connect (OSTI)

    Gong, Xiaojuan; Moghaddam, Minoo J.; Sagnella, Sharon M.; Conn, Charlotte E.; Danon, Stephen J.; Waddington, Lynne J.; Drummond, Calum J.

    2014-09-24

    An amphiphile prodrug, 5'-deoxy-5-fluoro-N{sup 4}-(palmityloxycarbonyl) cytidine or 5'-deoxy-5-fluoro-N{sup 4}-(hexadecanaloxycarbonyl) cytidine (5-FCPal), consisting of the same head group as the commercially available chemotherapeutic agent Capecitabine, linked to a palmityl hydrocarbon chain via a carbamate bond is reported. Thermal analysis of this prodrug indicates that it melts at {approx}115 C followed quickly by degradation beginning at {approx}120 C. The neat solid 5-FCPal amphiphile acquires a lamellar crystalline arrangement with a d-spacing of 28.6 {+-} 0.3 {angstrom}, indicating interdigitation of the hydrocarbon chains. Under aqueous conditions, solid 5-FCPal is non-swelling and no lyotropic liquid crystalline phase formation is observed. In order to assess the in vitro toxicity and in vivo efficacy in colloidal form, solid lipid nanoparticles (SLNs) with an average size of {approx}700 nm were produced via high pressure homogenization. The in vitro toxicity of the 5-FCPal SLNs against several different cancer and normal cell types was assessed over a 48 h period, and IC{sub 50} values were comparable to those observed for Capecitabine. The in vivo efficacy of the 5-FCPal SLNs was then assessed against the highly aggressive mouse 4T1 breast cancer model. To do so, the prodrug SLNs were administered orally at 3 different dosages (0.1, 0.25, 0.5 mmol/mouse/day) and compared to Capecitabine delivered at the same dosages. After 21 days of receiving the treatments, the 0.5 mmol dose of 5-FCPal exhibited the smallest average tumour volume. Since 5-FCPal is activated in a similar manner to Capecitabine via a 3 step enzymatic pathway with the final step occurring preferentially at the tumour site, formulation of the prodrug into SLNs combines the advantage of selective, localized activation with the sustained release properties of nanostructured amphiphile self-assembly and multiple payload materials thereby potentially creating a more effective anticancer agent.

  17. Estimation of placental and lactational transfer and tissue distribution of atrazine and its main metabolites in rodent dams, fetuses, and neonates with physiologically based pharmacokinetic modeling

    SciTech Connect (OSTI)

    Lin, Zhoumeng; Fisher, Jeffrey W.; Wang, Ran; Ross, Matthew K.; Filipov, Nikolay M.

    2013-11-15

    Atrazine (ATR) is a widely used chlorotriazine herbicide, a ubiquitous environmental contaminant, and a potential developmental toxicant. To quantitatively evaluate placental/lactational transfer and fetal/neonatal tissue dosimetry of ATR and its major metabolites, physiologically based pharmacokinetic models were developed for rat dams, fetuses and neonates. These models were calibrated using pharmacokinetic data from rat dams repeatedly exposed (oral gavage; 5 mg/kg) to ATR followed by model evaluation against other available rat data. Model simulations corresponded well to the majority of available experimental data and suggest that: (1) the fetus is exposed to both ATR and its major metabolite didealkylatrazine (DACT) at levels similar to maternal plasma levels, (2) the neonate is exposed mostly to DACT at levels two-thirds lower than maternal plasma or fetal levels, while lactational exposure to ATR is minimal, and (3) gestational carryover of DACT greatly affects its neonatal dosimetry up until mid-lactation. To test the model's cross-species extrapolation capability, a pharmacokinetic study was conducted with pregnant C57BL/6 mice exposed (oral gavage; 5 mg/kg) to ATR from gestational day 12 to 18. By using mouse-specific parameters, the model predictions fitted well with the measured data, including placental ATR/DACT levels. However, fetal concentrations of DACT were overestimated by the model (10-fold). This overestimation suggests that only around 10% of the DACT that reaches the fetus is tissue-bound. These rodent models could be used in fetal/neonatal tissue dosimetry predictions to help design/interpret early life toxicity/pharmacokinetic studies with ATR and as a foundation for scaling to humans. - Highlights: We developed PBPK models for atrazine in rat dams, fetuses, and neonates. We conducted pharmacokinetic (PK) study with atrazine in pregnant mice. Model predictions were in good agreement with experimental rat and mouse PK data. The fetus is exposed to atrazine/its main metabolite at levels similar to the dam. The nursing neonate is exposed primarily to atrazine's main metabolite DACT.

  18. Cyanidin-3-glucoside inhibits UVB-induced oxidative damage and inflammation by regulating MAP kinase and NF-κB signaling pathways in SKH-1 hairless mice skin

    SciTech Connect (OSTI)

    Pratheeshkumar, Poyil; Son, Young-Ok; Wang, Xin; Divya, Sasidharan Padmaja; Joseph, Binoy; Hitron, John Andrew; Wang, Lei; Kim, Donghern; Yin, Yuanqin; Roy, Ram Vinod; Lu, Jian; Zhang, Zhuo; Wang, Yitao; and others

    2014-10-01

    Skin cancer is one of the most commonly diagnosed cancers in the United States. Exposure to ultraviolet-B (UVB) radiation induces inflammation and photocarcinogenesis in mammalian skin. Cyanidin-3-glucoside (C3G), a member of the anthocyanin family, is present in various vegetables and fruits especially in edible berries, and displays potent antioxidant and anticarcinogenic properties. In this study, we have assessed the in vivo effects of C3G on UVB irradiation induced chronic inflammatory responses in SKH-1 hairless mice, a well-established model for UVB-induced skin carcinogenesis. Here, we show that C3G inhibited UVB-induced skin damage and inflammation in SKH-1 hairless mice. Our results indicate that C3G inhibited glutathione depletion, lipid peroxidation and myeloperoxidation in mouse skin by chronic UVB exposure. C3G significantly decreased the production of UVB-induced pro-inflammatory cytokines, such as IL-6 and TNF-α, associated with cutaneous inflammation. Likewise, UVB-induced inflammatory responses were diminished by C3G as observed by a remarkable reduction in the levels of phosphorylated MAP kinases, Erk1/2, p38, JNK1/2 and MKK4. Furthermore, C3G also decreased UVB-induced cyclooxygenase-2 (COX-2), PGE{sub 2} and iNOS levels, which are well-known key mediators of inflammation and cancer. Treatment with C3G inhibited UVB-induced nuclear translocation of NF-κB and degradation of IκBα in mice skin. Immunofluorescence assay revealed that topical application of C3G inhibited the expression of 8-hydroxy-2′-deoxyguanosine, proliferating cell nuclear antigen, and cyclin D1 in chronic UVB exposed mouse skin. Collectively, these data indicates that C3G can provide substantial protection against the adverse effects of UVB radiation by modulating UVB-induced MAP kinase and NF-κB signaling pathways. - Highlights: • C3G inhibited UVB-induced oxidative damage and inflammation. • C3G inhibited UVB-induced COX-2, iNOS and PGE{sub 2} production. • C3G inhibited UVB-induced elevated proinflammatory cytokine level. • C3G inhibited UVB-induced p38 MAP kinase signaling. • C3G inhibited UVB-induced NF-κB activation.

  19. Fibroblasts maintained in 3 dimensions show a better differentiation state and higher sensitivity to estrogens

    SciTech Connect (OSTI)

    Montani, Claudia; Steimberg, Nathalie; Boniotti, Jennifer; Biasiotto, Giorgio; Zanella, Isabella; Diafera, Giuseppe; Biunno, Ida; Caimi, Luigi; Mazzoleni, Giovanna; Di Lorenzo, Diego

    2014-11-01

    Cell differentiation and response to hormonal signals were studied in a 3D environment on an in-house generated mouse fibroblast cell line expressing a reporter gene under the control of estrogen responsive sequences (EREs). 3D cell culture conditions were obtained in a Rotary Cell Culture System; (RCCS™), a microgravity based bioreactor that promotes the aggregation of cells into multicellular spheroids (MCS). In this bioreactor the cells maintained a better differentiated phenotype and more closely resembled in vivo tissue. The RCCS™ cultured fibroblasts showed higher expression of genes regulating cell assembly, differentiation and hormonal functions. Microarray analysis showed that genes related to cell cycle, proliferation, cytoskeleton, migration, adhesion and motility were all down-regulated in 3D as compared to 2D conditions, as well as oncogene expression and inflammatory cytokines. Controlled remodeling of ECM, which is an essential aspect of cell organization, homeostasis and tissue was affected by the culture method as assessed by immunolocalization of β-tubulin. Markers of cell organization, homeostasis and tissue repair, metalloproteinase 2 (MMP2) and its physiological inhibitor (TIMP4) changed expression in association with the relative formation of cell aggregates. The fibroblasts cultured in the RCCS™ maintain a better responsiveness to estrogens, measured as expression of ERα and regulation of an ERE-dependent reporter and of the endogenous target genes CBP, Rarb, MMP1 and Dbp. Our data highlight the interest of this 3D culture model for its potential application in the field of cell response to hormonal signals and the pharmaco-toxicological analyses of chemicals and natural molecules endowed of estrogenic potential. - Highlights: • We here characterized the first cell line derived from an estrogen reporter mouse. • In the RCCS cells express an immortalized behavior but not a transformed phenotype. • The RCCS provides a system for maintaining cells in more physiological conditions. • RCCS-cultured fibroblasts showed higher hormonal sensitivity to estradiol. • This bioreactor is a novel 3D model to be applied to pharmacotoxicological studies.

  20. Case study on the utility of hepatic global gene expression profiling in the risk assessment of the carcinogen furan

    SciTech Connect (OSTI)

    Jackson, Anna Francina; Williams, Andrew; Recio, Leslie; Waters, Michael D.; Lambert, Iain B.; Yauk, Carole L.

    2014-01-01

    Furan is a chemical hepatocarcinogen in mice and rats. Its previously postulated cancer mode of action (MOA) is chronic cytotoxicity followed by sustained regenerative proliferation; however, its molecular basis is unknown. To this end, we conducted toxicogenomic analysis of B3C6F1 mouse livers following three week exposures to non-carcinogenic (0, 1, 2 mg/kg bw) or carcinogenic (4 and 8 mg/kg bw) doses of furan. We saw enrichment for pathways responsible for cytotoxicity: stress-activated protein kinase (SAPK) and death receptor (DR5 and TNF-alpha) signaling, and proliferation: extracellular signal-regulated kinases (ERKs) and TNF-alpha. We also noted the involvement of NF-kappaB and c-Jun in response to furan, which are genes that are known to be required for liver regeneration. Furan metabolism by CYP2E1 produces cis-2-butene-1,4-dial (BDA), which is required for ensuing cytotoxicity and oxidative stress. NRF2 is a master regulator of gene expression during oxidative stress and we suggest that chronic NFR2 activity and chronic inflammation may represent critical transition events between the adaptive (regeneration) and adverse (cancer) outcomes. Another objective of this study was to demonstrate the applicability of toxicogenomics data in quantitative risk assessment. We modeled benchmark doses for our transcriptional data and previously published cancer data, and observed consistency between the two. Margin of exposure values for both transcriptional and cancer endpoints were also similar. In conclusion, using furan as a case study we have demonstrated the value of toxicogenomics data in elucidating dose-dependent MOA transitions and in quantitative risk assessment. - Highlights: Global gene expression changes in furan-exposed mouse livers were analyzed. A molecular mode of action for furan-induced hepatocarcinogenesis is proposed. Key pathways include NRF2, SAPK, ERK and death receptor signaling. Important roles for TNF-alpha, c-Jun, and NF-?B in tumorigenesis are proposed. BMD and MoE values from transcriptional and apical data are compared.

  1. Pulmonary instillation of low doses of titanium dioxide nanoparticles in mice leads to particle retention and gene expression changes in the absence of inflammation

    SciTech Connect (OSTI)

    Husain, Mainul; Saber, Anne T.; Guo, Charles; Jacobsen, Nicklas R.; Jensen, Keld A.; Yauk, Carole L.; Williams, Andrew; Vogel, Ulla; Wallin, Hakan; Halappanavar, Sabina

    2013-06-15

    We investigated gene expression, protein synthesis, and particle retention in mouse lungs following intratracheal instillation of varying doses of nano-sized titanium dioxide (nano-TiO{sub 2}). Female C57BL/6 mice were exposed to rutile nano-TiO{sub 2} via single intratracheal instillations of 18, 54, and 162 μg/mouse. Mice were sampled 1, 3, and 28 days post-exposure. The deposition of nano-TiO{sub 2} in the lungs was assessed using nanoscale hyperspectral microscopy. Biological responses in the pulmonary system were analyzed using DNA microarrays, pathway-specific real-time RT-PCR (qPCR), gene-specific qPCR arrays, and tissue protein ELISA. Hyperspectral mapping showed dose-dependent retention of nano-TiO{sub 2} in the lungs up to 28 days post-instillation. DNA microarray analysis revealed approximately 3000 genes that were altered across all treatment groups (± 1.3 fold; p < 0.1). Several inflammatory mediators changed in a dose- and time-dependent manner at both the mRNA and protein level. Although no influx of neutrophils was detected at the low dose, changes in the expression of several genes and proteins associated with inflammation were observed. Resolving inflammation at the medium dose, and lack of neutrophil influx in the lung fluid at the low dose, were associated with down-regulation of genes involved in ion homeostasis and muscle regulation. Our gene expression results imply that retention of nano-TiO{sub 2} in the absence of inflammation over time may potentially perturb calcium and ion homeostasis, and affect smooth muscle activities. - Highlights: • Pulmonary effects following exposure to low doses of nano-TiO{sub 2} were examined. • Particle retention in lungs was assessed using nanoscale hyperspectral microscopy. • Particles persisted up to 28 days in lungs in all dose groups. • Inflammation was the pathway affected in the high dose group at all time points. • Ion homeostasis and muscle activity pathways were affected in the low dose group.

  2. Unexpected gender difference in sensitivity to the acute toxicity of dioxin in mice

    SciTech Connect (OSTI)

    Pohjanvirta, Raimo; Miettinen, Hanna; Sankari, Satu; Hegde, Nagabhooshan; Lindn, Jere; Department of Veterinary Biosciences, Faculty of Veterinary Medicine, University of Helsinki, P.O. Box 66, FI-00014 University of Helsinki

    2012-07-15

    The acute toxicity of the ubiquitous environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) varies widely among species and strains. Previous studies in rats have established that females are approximately 2-fold more sensitive to TCDD lethality than males. However, there is a surprising gap in the literature regarding possible gender-related sensitivity differences in mice. In the present study, by using three substrains of TCDD-sensitive C57BL/6 mice and transgenic mice on this background, we demonstrated that: 1) in contrast to the situation in rats, female mice are the more resistant gender; 2) the magnitude of the divergence between male and female mice depends on the substrain, but can amount to over 10-fold; 3) AH receptor protein expression levels or mutations in the primary structure of this receptor are not involved in the resistance of female mice of a C57BL/6 substrain, despite their acute LD{sub 50} for TCDD being over 5000 ?g/kg; 4) transgenic mice that globally express the rat wildtype AH receptor follow the mouse type of gender difference; 5) in gonadectomized mice, ovarian estrogens appear to enhance TCDD resistance, whereas testicular androgens seem to augment TCDD susceptibility; and 6) the gender difference correlates best with the severity of liver damage, which is also reflected in hepatic histopathology and the expression of pro-inflammatory cytokines, especially IL-6. Hence, the two closely related rodent species most often employed in toxicological risk characterization studies, rat and mouse, represent opposite examples of the influence of gender on dioxin sensitivity, further complicating the risk assessment of halogenated aromatic hydrocarbons. -- Highlights: ? In contrast to rats, male mice are more sensitive to TCDD toxicity than female mice. ? The resistance of female C57BL/6Kuo mice matches or exceeds that of male DBA/2 mice. ? The resistance of female C57BL/6Kuo mice is not based on AHR structure or abundance. ? Both androgens and estrogens appear to influence TCDD sensitivity. ? TCDD sensitivity correlates best with the severity of lesions in the liver.

  3. The non-competitive acetylcholinesterase inhibitor APS12-2 is a potent antagonist of skeletal muscle nicotinic acetylcholine receptors

    SciTech Connect (OSTI)

    Grandič, Marjana; Aráoz, Romulo; Molgó, Jordi; Turk, Tom; Sepčić, Kristina; Benoit, Evelyne; Frangež, Robert

    2012-12-01

    APS12-2, a non-competitive acetylcholinesterase inhibitor, is one of the synthetic analogs of polymeric alkylpyridinium salts (poly-APS) isolated from the marine sponge Reniera sarai. In the present work the effects of APS12-2 were studied on isolated mouse phrenic nerve–hemidiaphragm muscle preparations, using twitch tension measurements and electrophysiological recordings. APS12-2 in a concentration-dependent manner blocked nerve-evoked isometric muscle contraction (IC{sub 50} = 0.74 μM), without affecting directly-elicited twitch tension up to 2.72 μM. The compound (0.007–3.40 μM) decreased the amplitude of miniature endplate potentials until a complete block by concentrations higher than 0.68 μM, without affecting their frequency. Full size endplate potentials, recorded after blocking voltage-gated muscle sodium channels, were inhibited by APS12-2 in a concentration-dependent manner (IC{sub 50} = 0.36 μM) without significant change in the resting membrane potential of the muscle fibers up to 3.40 μM. The compound also blocked acetylcholine-evoked inward currents in Xenopus oocytes in which Torpedo (α1{sub 2}β1γδ) muscle-type nicotinic acetylcholine receptors (nAChRs) have been incorporated (IC{sub 50} = 0.0005 μM), indicating a higher affinity of the compound for Torpedo (α1{sub 2}β1γδ) than for the mouse (α1{sub 2}β1γε) nAChR. Our data show for the first time that APS12-2 blocks neuromuscular transmission by a non-depolarizing mechanism through an action on postsynaptic nAChRs of the skeletal neuromuscular junction. -- Highlights: ► APS12-2 produces concentration-dependent inhibition of nerve-evoked muscle contraction in vitro. ► APS12-2 blocks MEPPs and EPPs at the neuromuscular junction. APS12-2 blocks ACh-activated current in Xenopus oocytes incorporated with Torpedo nAChRs.

  4. Characterization of a double-sided silicon strip detector autoradiography system

    SciTech Connect (OSTI)

    Örbom, Anders Ahlstedt, Jonas; Östlund, Karl; Strand, Sven-Erik; Serén, Tom; Auterinen, Iiro; Kotiluoto, Petri; Hauge, Håvard; Olafsen, Tove; Wu, Anna M.; Dahlbom, Magnus

    2015-02-15

    Purpose: The most commonly used technology currently used for autoradiography is storage phosphor screens, which has many benefits such as a large field of view but lacks particle-counting detection of the time and energy of each detected radionuclide decay. A number of alternative designs, using either solid state or scintillator detectors, have been developed to address these issues. The aim of this study is to characterize the imaging performance of one such instrument, a double-sided silicon strip detector (DSSD) system for digital autoradiography. A novel aspect of this work is that the instrument, in contrast to previous prototype systems using the same detector type, provides the ability for user accessible imaging with higher throughput. Studies were performed to compare its spatial resolution to that of storage phosphor screens and test the implementation of multiradionuclide ex vivo imaging in a mouse preclinical animal study. Methods: Detector background counts were determined by measuring a nonradioactive sample slide for 52 h. Energy spectra and detection efficiency were measured for seven commonly used radionuclides under representative conditions for tissue imaging. System dead time was measured by imaging {sup 18}F samples of at least 5 kBq and studying the changes in count rate over time. A line source of {sup 58}Co was manufactured by irradiating a 10 μm nickel wire with fast neutrons in a research reactor. Samples of this wire were imaged in both the DSSD and storage phosphor screen systems and the full width at half maximum (FWHM) measured for the line profiles. Multiradionuclide imaging was employed in a two animal study to examine the intratumoral distribution of a {sup 125}I-labeled monoclonal antibody and a {sup 131}I-labeled engineered fragment (diabody) injected in the same mouse, both targeting carcinoembryonic antigen. Results: Detector background was 1.81 × 10{sup −6} counts per second per 50 × 50 μm pixel. Energy spectra and detection efficiency were successfully measured for seven radionuclides. The system dead time was measured to be 59 μs, and FWHM for a {sup 58}Co line source was 154 ± 14 μm for the DSSD system and 343 ± 15 μm for the storage phosphor system. Separation of the contributions from {sup 125}I and {sup 131}I was performed on autoradiography images of tumor sections. Conclusions: This study has shown that a DSSD system can be beneficially applied for digital autoradiography with simultaneous multiradionuclide imaging capability. The system has a low background signal, ability to image both low and high activity samples, and a good energy resolution.

  5. Role for DNA methylation in the regulation of miR-200c and miR-141 expression in normal and cancer cells

    SciTech Connect (OSTI)

    Vrba, Lukas; Jensen, Taylor J.; Garbe, James C.; Heimark, Ronald L.; Cress, Anne E.; Dickinson, Sally; Stampfer, Martha R.; Futscher, Bernard W.

    2009-12-23

    BACKGROUND: The microRNA-200 family participates in the maintenance of an epithelial phenotype and loss of its expression can result in epithelial to mesenchymal transition (EMT). Furthermore, the loss of expression of miR-200 family members is linked to an aggressive cancer phenotype. Regulation of the miR-200 family expression in normal and cancer cells is not fully understood. METHODOLOGY/ PRINCIPAL FINDINGS: Epigenetic mechanisms participate in the control of miR-200c and miR-141 expression in both normal and cancer cells. A CpG island near the predicted mir-200c/mir-141 transcription start site shows a striking correlation between miR-200c and miR-141 expression and DNA methylation in both normal and cancer cells, as determined by MassARRAY technology. The CpG island is unmethylated in human miR-200/miR-141 expressing epithelial cells and in miR-200c/miR-141 positive tumor cells. The CpG island is heavily methylated in human miR-200c/miR-141 negative fibroblasts and miR-200c/miR-141 negative tumor cells. Mouse cells show a similar inverse correlation between DNA methylation and miR-200c expression. Enrichment of permissive histone modifications, H3 acetylation and H3K4 trimethylation, is seen in normal miR-200c/miR-141-positive epithelial cells, as determined by chromatin immunoprecipitation coupled to real-time PCR. In contrast, repressive H3K9 dimethylation marks are present in normal miR-200c/miR-141-negative fibroblasts and miR-200c/miR-141 negative cancer cells and the permissive histone modifications are absent. The epigenetic modifier drug, 5-aza-2'-deoxycytidine, reactivates miR-200c/miR-141 expression showing that epigenetic mechanisms play a functional role in their transcriptional control. CONCLUSIONS/ SIGNIFICANCE: We report that DNA methylation plays a role in the normal cell type-specific expression of miR-200c and miR-141 and this role appears evolutionarily conserved, since similar results were obtained in mouse. Aberrant DNA methylation of the miR-200c/141 CpG island is closely linked to their inappropriate silencing in cancer cells. Since the miR-200c cluster plays a significant role in EMT, our results suggest an important role for DNA methylation in the control of phenotypic conversions in normal cells.

  6. MicroRNA-196b promotes cell proliferation and suppress cell differentiation in vitro

    SciTech Connect (OSTI)

    Cao, Donglin Hu, Liangshan; Lei, Da; Fang, Xiaolin; Zhang, Zhihong; Wang, Ting; Lin, Maorui; Huang, Jiwei; Yang, Huawen; Zhou, Xuan; Zhong, Limei

    2015-01-30

    Highlights: • miRNA-196b increases proliferation and blocks differentiation of progenitor cell. • miRNA-196b inhibits apoptosis and increases viability of cells lines. • Forced expression of miR-196b blocks the differentiation of THP1 induced by PMA. - Abstract: MicroRNA-196b (miR-196b) is frequently amplified and aberrantly overexpressed in acute leukemias. To investigate the role of miR-196b in acute leukemias, it has been observed that forced expression of this miRNA increases proliferation and inhibits apoptosis in human cell lines. More importantly, we show that this miRNA can significantly increase the colony-forming capacity of mouse normal bone marrow progenitor cells alone, as well as partially blocking the cells from differentiation. Taken together, our studies suggest that miRNA-196b may play an essential role in the development of MLL-associated leukemias through inhibiting cell differentiation and apoptosis, while promoting cell proliferation.

  7. The N-terminal leucine-zipper motif in PTRF/cavin-1 is essential and sufficient for its caveolae-association

    SciTech Connect (OSTI)

    Wei, Zhuang; Zou, Xinle; Wang, Hongzhong; Lei, Jigang; Wu, Yuan; Liao, Kan

    2015-01-16

    Highlight: • The N-terminal leucine-zipper motif in PTRF/cavin-1 determines caveolar association. • Different cellular localization of PTRF/cavin-1 influences its serine 389 and 391 phosphorylation state. • PTRF/cavin-1 regulates cell motility via its caveolar association. - Abstract: PTRF/cavin-1 is a protein of two lives. Its reported functions in ribosomal RNA synthesis and in caveolae formation happen in two different cellular locations: nucleus vs. plasma membrane. Here, we identified that the N-terminal leucine-zipper motif in PTRF/cavin-1 was essential for the protein to be associated with caveolae in plasma membrane. It could counteract the effect of nuclear localization sequence in the molecule (AA 235–251). Deletion of this leucine-zipper motif from PTRF/cavin-1 caused the mutant to be exclusively localized in nuclei. The fusion of this leucine-zipper motif with histone 2A, which is a nuclear protein, could induce the fusion protein to be exported from nucleus. Cell migration was greatly inhibited in PTRF/cavin-1{sup −/−} mouse embryonic fibroblasts (MEFs). The inhibited cell motility could only be rescued by exogenous cavin-1 but not the leucine-zipper motif deleted cavin-1 mutant. Plasma membrane dynamics is an important factor in cell motility control. Our results suggested that the membrane dynamics in cell migration is affected by caveolae associated PTRF/cavin-1.

  8. Detection of Weakly Conserved Ancestral Mammalian RegulatorySequences by Primate Comparisons

    SciTech Connect (OSTI)

    Wang, Qian-fei; Prabhakar, Shyam; Chanan, Sumita; Cheng,Jan-Fang; Rubin, Edward M.; Boffelli, Dario

    2006-06-01

    Genomic comparisons between human and distant, non-primatemammals are commonly used to identify cis-regulatory elements based onconstrained sequence evolution. However, these methods fail to detectcryptic functional elements, which are too weakly conserved among mammalsto distinguish from nonfunctional DNA. To address this problem, weexplored the potential of deep intra-primate sequence comparisons. Wesequenced the orthologs of 558 kb of human genomic sequence, coveringmultiple loci involved in cholesterol homeostasis, in 6 nonhumanprimates. Our analysis identified 6 noncoding DNA elements displayingsignificant conservation among primates, but undetectable in more distantcomparisons. In vitro and in vivo tests revealed that at least three ofthese 6 elements have regulatory function. Notably, the mouse orthologsof these three functional human sequences had regulatory activity despitetheir lack of significant sequence conservation, indicating that they arecryptic ancestral cis-regulatory elements. These regulatory elementscould still be detected in a smaller set of three primate speciesincluding human, rhesus and marmoset. Since the human and rhesus genomesequences are already available, and the marmoset genome is activelybeing sequenced, the primate-specific conservation analysis describedhere can be applied in the near future on a whole-genome scale, tocomplement the annotation provided by more distant speciescomparisons.

  9. ADAM15 expression is downregulated in melanoma metastasis compared to primary melanoma

    SciTech Connect (OSTI)

    Ungerer, Christopher; Doberstein, Kai; Boehm, Beate; Pfeilschifter, Josef; Mihic-Probst, Daniela; Gutwein, Paul

    2010-10-22

    Research highlights: {yields} Strong ADAM15 expression is found in normal melanocytes. {yields} ADAM15 expression is significantly downregulated in patients with melanoma metastasis. {yields} TGF-{beta} can downregulate ADAM15 expression in melanoma cells. {yields} Overexpression of ADAM15 in melanoma cells inhibits migration, proliferation and invasion of melanoma cells. {yields} Conclusion: ADAM15 represents an tumor suppressor protein in melanoma. -- Abstract: In a mouse melanoma metastasis model it has been recently shown that ADAM15 overexpression in melanoma cells significantly reduced the number of metastatic nodules on the lung. Unfortunately, the expression of ADAM15 in human melanoma tissue has not been determined so far. In our study, we characterized the expression of ADAM15 in tissue micro-arrays of patients with primary melanoma with melanoma metastasis. ADAM15 was expressed in melanocytes and endothelial cells of benign nevi and melanoma tissue. Importantly, ADAM15 was significantly downregulated in melanoma metastasis compared to primary melanoma. We further demonstrate that IFN-{gamma} and TGF-{beta} downregulate ADAM15 protein levels in melanoma cells. To investigate the role of ADAM15 in melanoma progression, we overexpressed ADAM15 in melanoma cells. Importantly, overexpression of ADAM15 in melanoma cells reduced the migration, invasion and the anchorage dependent and independent cell growth of melanoma cells. In summary, the downregulation of ADAM15 plays an important role in melanoma progression and ADAM15 act as a tumorsuppressor in melanoma.

  10. The mapping and differentiation of biological and environmental elemental signatures in the fossil remains of a 50 million year old bird

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Egerton, Victoria M.; Wogelius, Roy A.; Norell, Mark A.; Edwards, Nicholas P.; Sellers, William I.; Bergmann, Uwe; Sokaras, Dimosthenis; Alonso-Mori, Roberto; Ignatyev, Konstantin; van Veelen, Arjen; et al

    2015-01-22

    The preservation of fossils reflects the interplay of inorganic and organic chemical processes, which should be clearly differentiated to make interpretations about the biology of extinct organisms. A new coliiformes bird (mouse bird) from the ~50 million year old Green River Formation (Wyoming, USA) has here been analysed using synchrotron X-ray fluorescence and environmental scanning electron microscopy with an attached X-ray energy dispersive system (ESEM-EDS). The concentration and distribution of 16 elements (Si, P, S, Cl, K, Ca, Ti, Mg, Fe, Ni, Cu, Zn, As, Br, Ba, Hg) has been mapped for individual points on the sample. S, Cu andmore » Zn map distinctly within visibly preserved feathers and X-ray Absorption Spectroscopy (XAS) shows that S and Cu within the feathers are organically bound in a similar manner to modern feathers. The morphological preservation of the feathers, on both macro- and microscopic scales, is variable throughout the fossil and the differences in the lateral microfacies have resulted in a morphological preservation gradient. This study clearly differentiates endogenous organic remains from those representing exogenous overprinted geochemical precipitates and illustrates the chemical complexity of the overall taphonomic process.« less

  11. Identification of genetic loci that control mammary tumor susceptibility through the host microenvironment

    SciTech Connect (OSTI)

    Zhang, Pengju; Lo, Alvin; Huang, Yurong; Huang, Ge; Liang, Guozhou; Mott, Joni; Karpen, Gary H.; Blakely, Eleanor A.; Bissell, Mina J.; Barcellos-Hoff, Mary Helen; Snijders, Antoine M.; Mao, Jian-Hua

    2015-03-09

    The interplay between host genetics, tumor microenvironment and environmental exposure in cancer susceptibility remains poorly understood. Here we assessed the genetic control of stromal mediation of mammary tumor susceptibility to low dose ionizing radiation (LDIR) using backcrossed F1 into BALB/c (F1Bx) between cancer susceptible (BALB/c) and resistant (SPRET/EiJ) mouse strains. Tumor formation was evaluated after transplantation of non-irradiated Trp53-/- BALB/c mammary gland fragments into cleared fat pads of F1Bx hosts. Genome-wide linkage analysis revealed 2 genetic loci that constitute the baseline susceptibility via host microenvironment. However, once challenged with LDIR, we discovered 13 additional loci that were enriched for genes involved in cytokines, including TGF?1 signaling. Surprisingly, LDIR-treated F1Bx cohort significantly reduced incidence of mammary tumors from Trp53-/- fragments as well as prolonged tumor latency, compared to sham-treated controls. We demonstrated further that plasma levels of specific cytokines were significantly correlated with tumor latency. Using an ex vivo 3-D assay, we confirmed TGF?1 as a strong candidate for reduced mammary invasion in SPRET/EiJ, which could explain resistance of this strain to mammary cancer risk following LDIR. Our results open possible new avenues to understand mechanisms of genes operating via the stroma that affect cancer risk from external environmental exposures.

  12. Cul4A overexpression associated with Gli1 expression in malignant pleural mesothelioma

    SciTech Connect (OSTI)

    Yang, Yi -Lin; Ni, Jian; Hsu, Ping -Chih; Mao, Jian -Hua; Hsieh, David; Xu, Angela; Chan, Geraldine; Au, Alfred; Xu, Zhidong; Jablons, David M.; You, Liang

    2015-07-27

    Malignant pleural mesothelioma (mesothelioma) is a highly aggressive cancer without an effective treatment. Cul4A, a scaffold protein that recruits substrates for degradation, is amplified in several human cancers, including mesothelioma. We have recently shown that Cul4A plays an oncogenic role in vitro and in a mouse model. In this study, we analysed clinical mesothelioma tumours and found moderate to strong expression of Cul4A in 70.9% (51/72) of these tumours, as shown by immunohistochemistry. In 72.2% mesothelioma tumours with increased Cul4A copy number identified by fluorescence in situ hybridization analysis, Cul4A protein expression was moderate to strong. Similarly, Cul4A was overexpressed and Cul4A copy number was increased in human mesothelioma cell lines. Because Gli1 is highly expressed in human mesothelioma cells, we compared Cul4A and Gli1 expression in mesothelioma tumours and found their expression associated (P < 0.05, chi-square). In mesothelioma cell lines, inhibiting Cul4A by siRNA decreased Gli1 expression, suggesting that Gli1 expression is, at least in part, regulated by Cul4A in mesothelioma cells. Our results suggest a linkage between Cul4A and Gli1 expression in human mesothelioma.

  13. Metazoan Gene Families from Metazome

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Metazome is a joint project of the Department of Energy's Joint Genome Institute and the Center for Integrative Genomics to facilitate comparative genomic studies amongst metazoans. Clusters of orthologous and paralogous genes that represent the modern descendents of ancestral gene sets are constructed at key phylogenetic nodes. These clusters allow easy access to clade specific orthology/paralogy relationships as well as clade specific genes and gene expansions. As of version 2.0.4, Metazome provides access to twenty-four sequenced and annotated metazoan genomes, clustered at nine evolutionarily significant nodes. Where possible, each gene has been annotated with PFAM, KOG, KEGG, and PANTHER assignments, and publicly available annotations from RefSeq, UniProt, Ensembl, and JGI are hyper-linked and searchable. The included organisms (by common name) are: Human, Mouse, Rat, Dog, Opossum, Chicken, Frog, Stickleback, Medaka, Fugu pufferfish; Zebrafish, Seasquirt - savignyi, Seasquirt - intestinalis, Amphioxus, Sea Urchin, Fruitfly, Mosquite, Yellow Fever Mosquito, Silkworm, Red Flour Beetle, Worm, Briggsae Worm, Owl limpet (snail), and Sea anemone. [Copied from Metazome Overview at http://www.metazome.net/Metazome_info.php

  14. cAMP-response-element-binding protein positively regulates breast cancer metastasis and subsequent bone destruction

    SciTech Connect (OSTI)

    Son, Jieun; Lee, Jong-Ho; Kim, Ha-Neui; Ha, Hyunil Lee, Zang Hee

    2010-07-23

    Research highlights: {yields} CREB is highly expressed in advanced breast cancer cells. {yields} Tumor-related factors such as TGF-{beta} further elevate CREB expression. {yields} CREB upregulation stimulates metastatic potential of breast cancer cells. {yields} CREB signaling is required for breast cancer-induced bone destruction. -- Abstract: cAMP-response-element-binding protein (CREB) signaling has been reported to be associated with cancer development and poor clinical outcome in various types of cancer. However, it remains to be elucidated whether CREB is involved in breast cancer development and osteotropism. Here, we found that metastatic MDA-MB-231 breast cancer cells exhibited higher CREB expression than did non-metastatic MCF-7 cells and that CREB expression was further increased by several soluble factors linked to cancer progression, such as IL-1, IGF-1, and TGF-{beta}. Using wild-type CREB and a dominant-negative form (K-CREB), we found that CREB signaling positively regulated the proliferation, migration, and invasion of MDA-MB-231 cells. In addition, K-CREB prevented MDA-MB-231 cell-induced osteolytic lesions in a mouse model of cancer metastasis. Furthermore, CREB signaling in cancer cells regulated the gene expression of PTHrP, MMPs, and OPG, which are closely involved in cancer metastasis and bone destruction. These results indicate that breast cancer cells acquire CREB overexpression during their development and that this CREB upregulation plays an important role in multiple steps of breast cancer bone metastasis.

  15. Numerical calculation of protein-ligand binding rates through solution of the Smoluchowski equation using smoothed particle hydrodynamics

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Pan, Wenxiao; Daily, Michael; Baker, Nathan A.

    2015-05-07

    Background: The calculation of diffusion-controlled ligand binding rates is important for understanding enzyme mechanisms as well as designing enzyme inhibitors. Methods: We demonstrate the accuracy and effectiveness of a Lagrangian particle-based method, smoothed particle hydrodynamics (SPH), to study diffusion in biomolecular systems by numerically solving the time-dependent Smoluchowski equation for continuum diffusion. Unlike previous studies, a reactive Robin boundary condition (BC), rather than the absolute absorbing (Dirichlet) BC, is considered on the reactive boundaries. This new BC treatment allows for the analysis of enzymes with “imperfect” reaction rates. Results: The numerical method is first verified in simple systems and thenmore » applied to the calculation of ligand binding to a mouse acetylcholinesterase (mAChE) monomer. Rates for inhibitor binding to mAChE are calculated at various ionic strengths and compared with experiment and other numerical methods. We find that imposition of the Robin BC improves agreement between calculated and experimental reaction rates. Conclusions: Although this initial application focuses on a single monomer system, our new method provides a framework to explore broader applications of SPH in larger-scale biomolecular complexes by taking advantage of its Lagrangian particle-based nature.« less

  16. Understanding dynamic changes in live cell adhesion with neutron reflectometry

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Junghans, Ann; Waltman, Mary Jo; Smith, Hillary L.; Pocivavsek, Luka; Zebda, Noureddine; Birukov, Konstantin; Viapiano, Mariano; Majewski, Jaroslaw

    2014-12-10

    In this study, neutron reflectometry (NR) was used to examine various live cells' adhesion to quartz substrates under different environmental conditions, including flow stress. To the best of our knowledge, these measurements represent the first successful visualization and quantization of the interface between live cells and a substrate with sub-nanometer resolution. In our first experiments, we examined live mouse fibroblast cells as opposed to past experiments using supported lipids, proteins, or peptide layers with no associated cells. We continued the NR studies of cell adhesion by investigating endothelial monolayers and glioblastoma cells under dynamic flow conditions. We demonstrated that neutronmore » reflectometry is a powerful tool to study the strength of cellular layer adhesion in living tissues, which is a key factor in understanding the physiology of cell interactions and conditions leading to abnormal or disease circumstances. Continuative measurements, such as investigating changes in tumor cell — surface contact of various glioblastomas, could impact advancements in tumor treatments. In principle, this can help us to identify changes that correlate with tumor invasiveness. Pursuit of these studies can have significant medical impact on the understanding of complex biological problems and their effective treatment, e.g. for the development of targeted anti-invasive therapies.« less

  17. Chemical Concentrations in Field Mice from Open-Detonation Firing Sites TA-36 Minie and TA-39 Point 6 at Los Alamos National Laboratory

    SciTech Connect (OSTI)

    Fresquez, Philip R.

    2011-01-01

    Field mice (mostly Peromyscus spp.) were collected at two open-detonation (high explosive) firing sites - Minie at Technical Area (TA) 36 and Point 6 at TA-39 - at Los Alamos National Laboratory in August of 2010 and in February of 2011 for chemical analysis. Samples of whole body field mice from both sites were analyzed for target analyte list elements (mostly metals), dioxin/furans, polychlorinated biphenyl congeners, high explosives, and perchlorate. In addition, uranium isotopes were analyzed in a composite sample collected from TA-36 Minie. In general, all constituents, with the exception of lead at TA-39 Point 6, in whole body field mice samples collected from these two open-detonation firing sites were either not detected or they were detected below regional statistical reference levels (99% confidence level), biota dose screening levels, and/or soil ecological chemical screening levels. The amount of lead in field mice tissue collected from TA-39 Point 6 was higher than regional background, and some lead levels in the soil were higher than the ecological screening level for the field mouse; however, these levels are not expected to affect the viability of the populations over the site as a whole.

  18. New insights into potential functions for the protein 4.1superfamily of proteins in kidney epithelium

    SciTech Connect (OSTI)

    Calinisan, Venice; Gravem, Dana; Chen, Ray Ping-Hsu; Brittin,Sachi; Mohandas, Narla; Lecomte, Marie-Christine; Gascard, Philippe

    2005-06-17

    Members of the protein 4.1 family of adapter proteins are expressed in a broad panel of tissues including various epithelia where they likely play an important role in maintenance of cell architecture and polarity and in control of cell proliferation. We have recently characterized the structure and distribution of three members of the protein 4.1 family, 4.1B, 4.1R and 4.1N, in mouse kidney. We describe here binding partners for renal 4.1 proteins, identified through the screening of a rat kidney yeast two-hybrid system cDNA library. The identification of putative protein 4.1-based complexes enables us to envision potential functions for 4.1 proteins in kidney: organization of signaling complexes, response to osmotic stress, protein trafficking, and control of cell proliferation. We discuss the relevance of these protein 4.1-based interactions in kidney physio-pathology in the context of their previously identified functions in other cells and tissues. Specifically, we will focus on renal 4.1 protein interactions with beta amyloid precursor protein (beta-APP), 14-3-3 proteins, and the cell swelling-activated chloride channel pICln. We also discuss the functional relevance of another member of the protein 4.1 superfamily, ezrin, in kidney physiopathology.

  19. Targeted deletion of Kif18a protects from colitis-associated colorectal (CAC) tumors in mice through impairing Akt phosphorylation

    SciTech Connect (OSTI)

    Zhu, Houbao [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China)] [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China); Xu, Wangyang [Department of Clinical Laboratories, Ninth Peoples Hospital, SJTUSM, Shanghai 200011 (China)] [Department of Clinical Laboratories, Ninth Peoples Hospital, SJTUSM, Shanghai 200011 (China); Zhang, Hongxin [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China)] [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China); Liu, Jianbing [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China) [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China); Shanghai Research Center for Model Organisms, Shanghai 201203 (China); Xu, Haimin [Department of Pathology, Rui-Jin Hospital, SJTUSM, Shanghai 200025 (China)] [Department of Pathology, Rui-Jin Hospital, SJTUSM, Shanghai 200025 (China); Lu, Shunyuan; Dang, Suying [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China)] [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China); Kuang, Ying [Shanghai Research Center for Model Organisms, Shanghai 201203 (China)] [Shanghai Research Center for Model Organisms, Shanghai 201203 (China); Jin, Xiaolong [Department of Pathology, Rui-Jin Hospital, SJTUSM, Shanghai 200025 (China)] [Department of Pathology, Rui-Jin Hospital, SJTUSM, Shanghai 200025 (China); Wang, Zhugang, E-mail: zhugangw@shsmu.edu.cn [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China) [State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital and Department of Medical Genetics, E-Institutes of Shanghai Universities, Shanghai Jiao Tong University School of Medicine (SJTUSM), Shanghai 200025 (China); Shanghai Research Center for Model Organisms, Shanghai 201203 (China)

    2013-08-16

    Highlights: Kif18A is up-regulated in CAC of mouse model. Kif18a{sup ?/?} mice are protected from CAC. Tumor cells from Kif18a{sup ?/?} mice undergo more apoptosis. Kif18A deficiency induces poor Atk phosphorylation. -- Abstract: Kinesins are a superfamily of molecular motors involved in cell division or intracellular transport. They are becoming important targets for chemotherapeutic intervention of cancer due to their crucial role in mitosis. Here, we demonstrate that the kinesin-8 Kif18a is overexpressed in murine CAC and is a crucial promoter during early CAC carcinogenesis. Kif18a-deficient mice are evidently protected from AOMDSS-induced colon carcinogenesis. Kif18A is responsible for proliferation of colonic tumor cells, while Kif18a ablation in mice promotes cell apoptosis. Mechanistically, Kif18a is responsible for induction of Akt phosphorylation, which is known to be associated with cell survival regulation. In conclusion, Kif18a is critical for colorectal carcinogenesis in the setting of inflammation by mechanisms of increased PI3K-AKT signaling. Inhibition of Kif18A activity may be useful in the prevention or chemotherapeutic intervention of CAC.

  20. Mechanisms of radiation-induced gene responses

    SciTech Connect (OSTI)

    Woloschak, G.E.; Paunesku, T.

    1996-10-01

    In the process of identifying genes differentially expressed in cells exposed ultraviolet radiation, we have identified a transcript having a 26-bp region that is highly conserved in a variety of species including Bacillus circulans, yeast, pumpkin, Drosophila, mouse, and man. When the 5` region (flanking region or UTR) of a gene, the sequence is predominantly in +/+ orientation with respect to the coding DNA strand; while in the coding region and the 3` region (UTR), the sequence is most frequently in the +/-orientation with respect to the coding DNA strand. In two genes, the element is split into two parts; however, in most cases, it is found only once but with a minimum of 11 consecutive nucleotides precisely depicting the original sequence. The element is found in a large number of different genes with diverse functions (from human ras p21 to B. circulans chitonase). Gel shift assays demonstrated the presence of a protein in HeLa cell extracts that binds to the sense and antisense single-stranded consensus oligomers, as well as to the double- stranded oligonucleotide. When double-stranded oligomer was used, the size shift demonstrated as additional protein-oligomer complex larger than the one bound to either sense or antisense single-stranded consensus oligomers alone. It is speculated either that this element binds to protein(s) important in maintaining DNA is a single-stranded orientation for transcription or, alternatively that this element is important in the transcription-coupled DNA repair process.

  1. Experimental comparison of grating- and propagation-based hard X-ray phase tomography of soft tissue

    SciTech Connect (OSTI)

    Lang, S.; Schulz, G.; Müller, B.; Zanette, I.; Dominietto, M.; Langer, M.; Rack, A.; Le Duc, G.; David, C.; Mohr, J.; Pfeiffer, F.; Weitkamp, T.

    2014-10-21

    When imaging soft tissues with hard X-rays, phase contrast is often preferred over conventional attenuation contrast due its superior sensitivity. However, it is unclear which of the numerous phase tomography methods yields the optimized results at given experimental conditions. Therefore, we quantitatively compared the three phase tomography methods implemented at the beamline ID19 of the European Synchrotron Radiation Facility: X-ray grating interferometry (XGI), and propagation-based phase tomography, i.e., single-distance phase retrieval (SDPR) and holotomography (HT), using cancerous tissue from a mouse model and an entire heart of a rat. We show that for both specimens, the spatial resolution derived from the characteristic morphological features is about a factor of two better for HT and SDPR compared to XGI, whereas the XGI data generally exhibit much better contrast-to-noise ratios for the anatomical features. Moreover, XGI excels in fidelity of the density measurements, and is also more robust against low-frequency artifacts than HT, but it might suffer from phase-wrapping artifacts. Thus, we can regard the three phase tomography methods discussed as complementary. The application will decide which spatial and density resolutions are desired, for the imaging task and dose requirements, and, in addition, the applicant must choose between the complexity of the experimental setup and the one of data processing.

  2. Fast kinase domain-containing protein 3 is a mitochondrial protein essential for cellular respiration

    SciTech Connect (OSTI)

    Simarro, Maria; Gimenez-Cassina, Alfredo; Kedersha, Nancy; Lazaro, Jean-Bernard; Adelmant, Guillaume O.; Marto, Jarrod A.; Rhee, Kirsten; Tisdale, Sarah; Danial, Nika; Benarafa, Charaf; Orduna, Anonio; Anderson, Paul

    2010-10-22

    Research highlights: {yields} Five members of the FAST kinase domain-containing proteins are localized to mitochondria in mammalian cells. {yields} The FASTKD3 interactome includes proteins involved in various aspects of mitochondrial metabolism. {yields} Targeted knockdown of FASTKD3 significantly reduces basal and maximal mitochondrial oxygen consumption. -- Abstract: Fas-activated serine/threonine phosphoprotein (FAST) is the founding member of the FAST kinase domain-containing protein (FASTKD) family that includes FASTKD1-5. FAST is a sensor of mitochondrial stress that modulates protein translation to promote the survival of cells exposed to adverse conditions. Mutations in FASTKD2 have been linked to a mitochondrial encephalomyopathy that is associated with reduced cytochrome c oxidase activity, an essential component of the mitochondrial electron transport chain. We have confirmed the mitochondrial localization of FASTKD2 and shown that all FASTKD family members are found in mitochondria. Although human and mouse FASTKD1-5 genes are expressed ubiquitously, some of them are most abundantly expressed in mitochondria-enriched tissues. We have found that RNA interference-mediated knockdown of FASTKD3 severely blunts basal and stress-induced mitochondrial oxygen consumption without disrupting the assembly of respiratory chain complexes. Tandem affinity purification reveals that FASTKD3 interacts with components of mitochondrial respiratory and translation machineries. Our results introduce FASTKD3 as an essential component of mitochondrial respiration that may modulate energy balance in cells exposed to adverse conditions by functionally coupling mitochondrial protein synthesis to respiration.

  3. The mapping and differentiation of biological and environmental elemental signatures in the fossil remains of a 50 million year old bird

    SciTech Connect (OSTI)

    Egerton, Victoria M.; Wogelius, Roy A.; Norell, Mark A.; Edwards, Nicholas P.; Sellers, William I.; Bergmann, Uwe; Sokaras, Dimosthenis; Alonso-Mori, Roberto; Ignatyev, Konstantin; van Veelen, Arjen; Anné, Jennifer; van Dongen, Bart; Knoll, Fabien; Manning, Phillip L.

    2015-01-22

    The preservation of fossils reflects the interplay of inorganic and organic chemical processes, which should be clearly differentiated to make interpretations about the biology of extinct organisms. A new coliiformes bird (mouse bird) from the ~50 million year old Green River Formation (Wyoming, USA) has here been analysed using synchrotron X-ray fluorescence and environmental scanning electron microscopy with an attached X-ray energy dispersive system (ESEM-EDS). The concentration and distribution of 16 elements (Si, P, S, Cl, K, Ca, Ti, Mg, Fe, Ni, Cu, Zn, As, Br, Ba, Hg) has been mapped for individual points on the sample. S, Cu and Zn map distinctly within visibly preserved feathers and X-ray Absorption Spectroscopy (XAS) shows that S and Cu within the feathers are organically bound in a similar manner to modern feathers. The morphological preservation of the feathers, on both macro- and microscopic scales, is variable throughout the fossil and the differences in the lateral microfacies have resulted in a morphological preservation gradient. This study clearly differentiates endogenous organic remains from those representing exogenous overprinted geochemical precipitates and illustrates the chemical complexity of the overall taphonomic process.

  4. Accelerated Evolution of Conserved Noncoding Sequences in theHuman Genome

    SciTech Connect (OSTI)

    Prambhakar, Shyam; Noonan, James P.; Paabo, Svante; Rubin, EdwardM.

    2006-07-06

    Genomic comparisons between human and distant, non-primatemammals are commonly used to identify cis-regulatory elements based onconstrained sequence evolution. However, these methods fail to detect"cryptic" functional elements, which are too weakly conserved amongmammals to distinguish from nonfunctional DNA. To address this problem,we explored the potential of deep intra-primate sequence comparisons. Wesequenced the orthologs of 558 kb of human genomic sequence, coveringmultiple loci involved in cholesterol homeostasis, in 6 nonhumanprimates. Our analysis identified 6 noncoding DNA elements displayingsignificant conservation among primates, but undetectable in more distantcomparisons. In vitro and in vivo tests revealed that at least three ofthese 6 elements have regulatory function. Notably, the mouse orthologsof these three functional human sequences had regulatory activity despitetheir lack of significant sequence conservation, indicating that they arecryptic ancestral cis-regulatory elements. These regulatory elementscould still be detected in a smaller set of three primate speciesincluding human, rhesus and marmoset. Since the human and rhesus genomesequences are already available, and the marmoset genome is activelybeing sequenced, the primate-specific conservation analysis describedhere can be applied in the near future on a whole-genome scale, tocomplement the annotation provided by more distant speciescomparisons.

  5. In Vivo Enhancer Analysis Chromosome 16 Conserved NoncodingSequences

    SciTech Connect (OSTI)

    Pennacchio, Len A.; Ahituv, Nadav; Moses, Alan M.; Nobrega,Marcelo; Prabhakar, Shyam; Shoukry, Malak; Minovitsky, Simon; Visel,Axel; Dubchak, Inna; Holt, Amy; Lewis, Keith D.; Plajzer-Frick, Ingrid; Akiyama, Jennifer; De Val, Sarah; Afzal, Veena; Black, Brian L.; Couronne, Olivier; Eisen, Michael B.; Rubin, Edward M.

    2006-02-01

    The identification of enhancers with predicted specificitiesin vertebrate genomes remains a significant challenge that is hampered bya lack of experimentally validated training sets. In this study, weleveraged extreme evolutionary sequence conservation as a filter toidentify putative gene regulatory elements and characterized the in vivoenhancer activity of human-fish conserved and ultraconserved1 noncodingelements on human chromosome 16 as well as such elements from elsewherein the genome. We initially tested 165 of these extremely conservedsequences in a transgenic mouse enhancer assay and observed that 48percent (79/165) functioned reproducibly as tissue-specific enhancers ofgene expression at embryonic day 11.5. While driving expression in abroad range of anatomical structures in the embryo, the majority of the79 enhancers drove expression in various regions of the developingnervous system. Studying a set of DNA elements that specifically droveforebrain expression, we identified DNA signatures specifically enrichedin these elements and used these parameters to rank all ~;3,400human-fugu conserved noncoding elements in the human genome. The testingof the top predictions in transgenic mice resulted in a three-foldenrichment for sequences with forebrain enhancer activity. These datadramatically expand the catalogue of in vivo-characterized human geneenhancers and illustrate the future utility of such training sets for avariety of iological applications including decoding the regulatoryvocabulary of the human genome.

  6. Predicting Tissue-Specific Enhancers in the Human Genome

    SciTech Connect (OSTI)

    Pennacchio, Len A.; Loots, Gabriela G.; Nobrega, Marcelo A.; Ovcharenko, Ivan

    2006-07-01

    Determining how transcriptional regulatory signals areencoded in vertebrate genomes is essential for understanding the originsof multi-cellular complexity; yet the genetic code of vertebrate generegulation remains poorly understood. In an attempt to elucidate thiscode, we synergistically combined genome-wide gene expression profiling,vertebrate genome comparisons, and transcription factor binding siteanalysis to define sequence signatures characteristic of candidatetissue-specific enhancers in the human genome. We applied this strategyto microarray-based gene expression profiles from 79 human tissues andidentified 7,187 candidate enhancers that defined their flanking geneexpression, the majority of which were located outside of knownpromoters. We cross-validated this method for its ability to de novopredict tissue-specific gene expression and confirmed its reliability in57 of the 79 available human tissues, with an average precision inenhancer recognition ranging from 32 percent to 63 percent, and asensitivity of 47 percent. We used the sequence signatures identified bythis approach to assign tissue-specific predictions to ~;328,000human-mouse conserved noncoding elements in the human genome. Byoverlapping these genome-wide predictions with a large in vivo dataset ofenhancers validated in transgenic mice, we confirmed our results with a28 percent sensitivity and 50 percent precision. These results indicatethe power of combining complementary genomic datasets as an initialcomputational foray into the global view of tissue-specific generegulation in vertebrates.

  7. Two Rounds of Whole Genome Duplication in the AncestralVertebrate

    SciTech Connect (OSTI)

    Dehal, Paramvir; Boore, Jeffrey L.

    2005-04-12

    The hypothesis that the relatively large and complex vertebrate genome was created by two ancient, whole genome duplications has been hotly debated, but remains unresolved. We reconstructed the evolutionary relationships of all gene families from the complete gene sets of a tunicate, fish, mouse, and human, then determined when each gene duplicated relative to the evolutionary tree of the organisms. We confirmed the results of earlier studies that there remains little signal of these events in numbers of duplicated genes, gene tree topology, or the number of genes per multigene family. However, when we plotted the genomic map positions of only the subset of paralogous genes that were duplicated prior to the fish-tetrapod split, their global physical organization provides unmistakable evidence of two distinct genome duplication events early in vertebrate evolution indicated by clear patterns of 4-way paralogous regions covering a large part of the human genome. Our results highlight the potential for these large-scale genomic events to have driven the evolutionary success of the vertebrate lineage.

  8. MCM ring hexamerization is a prerequisite for DNA-binding

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Froelich, Clifford A.; Nourse, Amanda; Enemark, Eric J.

    2015-09-13

    The hexameric Minichromosome Maintenance (MCM) protein complex forms a ring that unwinds DNA at the replication fork in eukaryotes and archaea. Our recent crystal structure of an archaeal MCM N-terminal domain bound to single-stranded DNA (ssDNA) revealed ssDNA associating across tight subunit interfaces but not at the loose interfaces, indicating that DNA-binding is governed not only by the DNA-binding residues of the subunits (MCM ssDNA-binding motif, MSSB) but also by the relative orientation of the subunits. We now extend these findings to show that DNA-binding by the MCM N-terminal domain of the archaeal organism Pyrococcus furiosus occurs specifically in themore » hexameric oligomeric form. We show that mutants defective for hexamerization are defective in binding ssDNA despite retaining all the residues observed to interact with ssDNA in the crystal structure. One mutation that exhibits severely defective hexamerization and ssDNA-binding is at a conserved phenylalanine that aligns with the mouse Mcm4(Chaos3) mutation associated with chromosomal instability, cancer, and decreased intersubunit association.« less

  9. Non-cell-autonomous stimulation of stem cell proliferation following ablation of Tcf3

    SciTech Connect (OSTI)

    Yi, Fei; Merrill, Bradley J.

    2010-04-01

    A combination of cell intrinsic factors and extracellular signals determine whether mouse embryonic stem cells (ESC) divide, self-renew, and differentiate. Here, we report a new interaction between cell intrinsic aspects of the canonical Wnt/Tcf/{beta}-catenin signaling pathway and extracellular Lif/Jak/Stat3 stimulation that combines to promote self-renewal and proliferation of ESC. Mutant ESC lacking the Tcf3 transcriptional repressor continue to self-renew in the absence of exogenous Lif and through pharmacological inhibition of Lif/Jak/Stat3 signaling; however, proliferation rates of TCF3-/- ESC were significantly decreased by inhibiting Jak/Stat3 activity. Cell mixing experiments showed that stimulation of Stat3 phosphorylation in TCF3-/- ESC was mediated through secretion of paracrine acting factors, but did not involve elevated Lif or LifR transcription. The new interaction between Wnt and Lif/Jak/Stat3 signaling pathways has potential for new insights into the growth of tumors caused by aberrant activity of Wnt/Tcf/{beta}-catenin signaling.

  10. Cell recognition molecule L1 promotes embryonic stem cell differentiation through the regulation of cell surface glycosylation

    SciTech Connect (OSTI)

    Li, Ying; Department of Clinical Laboratory, Second Affiliated Hospital of Dalian Medical University, Dalian 116023 ; Huang, Xiaohua; Department of Clinical Biochemistry, College of Laboratory Medicine, Dalian Medical University, Dalian 116044 ; An, Yue; Ren, Feng; Yang, Zara Zhuyun; Zhu, Hongmei; Zhou, Lei; Department of Anatomy and Developmental Biology, Monash University, Clayton 3800 ; He, Xiaowen; Schachner, Melitta; Xiao, Zhicheng; Department of Anatomy and Developmental Biology, Monash University, Clayton 3800 ; Ma, Keli; Li, Yali; Department of Anatomy, National University of Singapore, Singapore 119078

    2013-10-25

    Highlights: Down-regulating FUT9 and ST3Gal4 expression blocks L1-induced neuronal differentiation of ESCs. Up-regulating FUT9 and ST3Gal4 expression in L1-ESCs depends on the activation of PLC?. L1 promotes ESCs to differentiate into neuron through regulating cell surface glycosylation. -- Abstract: Cell recognition molecule L1 (CD171) plays an important role in neuronal survival, migration, differentiation, neurite outgrowth, myelination, synaptic plasticity and regeneration after injury. Our previous study has demonstrated that overexpressing L1 enhances cell survival and proliferation of mouse embryonic stem cells (ESCs) through promoting the expression of FUT9 and ST3Gal4, which upregulates cell surface sialylation and fucosylation. In the present study, we examined whether sialylation and fucosylation are involved in ESC differentiation through L1 signaling. RNA interference analysis showed that L1 enhanced differentiation of ESCs into neurons through the upregulation of FUT9 and ST3Gal4. Furthermore, blocking the phospholipase C? (PLC?) signaling pathway with either a specific PLC? inhibitor or knockdown PLC? reduced the expression levels of both FUT9 and ST3Gal4 mRNAs and inhibited L1-mediated neuronal differentiation. These results demonstrate that L1 promotes neuronal differentiation from ESCs through the L1-mediated enhancement of FUT9 and ST3Gal4 expression.

  11. A system for combined three-dimensional morphological and molecular analysis of thick tissue specimens

    SciTech Connect (OSTI)

    Fernandez-Gonzalez, Rodrigo; Jones, Arthur; Garcia-Rodriguez, Enrique; Yuan Chen, Ping; Idica, Adam; Lockett, Stephen J.; Barcellos-Hoff, Mary Helen; Ortiz-de-Solorzano, Carlos

    2002-04-25

    We present a new system for simultaneous morphological and molecular analysis of thick tissue samples. The system is composed of a computer assisted microscope and a JAVA-based image display, analysis and visualization program that allows acquisition, annotation, meaningful storage, three-dimensional reconstruction and analysis of structures of interest in thick sectioned tissue specimens. We describe the system in detail and illustrate its use by imaging, reconstructing and analyzing two complete tissue blocks which were differently processed and stained. One block was obtained from a ductal carcinoma in situ (DCIS) lumpectomy specimen and stained alternatively with Hematoxilyn and Eosin (H&E), and with a counterstain and fluorescence in situ hybridization (FISH) to the ERB-B2 gene. The second block contained a fully sectioned mammary gland of a mouse, stained for Histology with H&E. We show how the system greatly reduces the amount of interaction required for the acquisition and analysis and is therefore suitable for studies that require morphologically driven, wide scale (e.g., whole gland) analysis of complex tissue samples or cultures.

  12. A Semiautomated Framework for Integrating Expert Knowledge into Disease Marker Identification

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wang, Jing; Webb-Robertson, Bobbie-Jo M.; Matzke, Melissa M.; Varnum, Susan M.; Brown, Joseph N.; Riensche, Roderick M.; Adkins, Joshua N.; Jacobs, Jon M.; Hoidal, John R.; Scholand, Mary Beth; et al

    2013-01-01

    Background . The availability of large complex data sets generated by high throughput technologies has enabled the recent proliferation of disease biomarker studies. However, a recurring problem in deriving biological information from large data sets is how to best incorporate expert knowledge into the biomarker selection process. Objective . To develop a generalizable framework that can incorporate expert knowledge into data-driven processes in a semiautomated way while providing a metric for optimization in a biomarker selection scheme. Methods . The framework was implemented as a pipeline consisting of five components for the identification of signatures from integrated clustering (ISIC).more » Expert knowledge was integrated into the biomarker identification process using the combination of two distinct approaches; a distance-based clustering approach and an expert knowledge-driven functional selection. Results . The utility of the developed framework ISIC was demonstrated on proteomics data from a study of chronic obstructive pulmonary disease (COPD). Biomarker candidates were identified in a mouse model using ISIC and validated in a study of a human cohort. Conclusions . Expert knowledge can be introduced into a biomarker discovery process in different ways to enhance the robustness of selected marker candidates. Developing strategies for extracting orthogonal and robust features from large data sets increases the chances of success in biomarker identification.« less

  13. Molecular Imaging of Conscious, Unrestrained Mice with AwakeSPECT

    SciTech Connect (OSTI)

    Baba, Justin S.; Endres, Christopher J.; Foss, Catherine A.; Nimmagadda, Sridhar; Jung, Hyeyun; Goddard, James S.; Lee, Seung Joon; McKisson, John; Smith, Mark F.; Stolin, Alexander V.; Weisenberger, Andrew G.; Pomper, Martin G.

    2013-06-01

    We have developed a SPECT imaging system, AwakeSPECT, to enable molecular brain imaging of untrained mice that are conscious, unanesthetized, and unrestrained. We accomplished this with head tracking and motion correction techniques. Methods: The capability of the system for motion-corrected imaging was demonstrated with a ^99mTc-pertechnetate phantom, ^99mTc-methylene diphosphonate bone imaging, and measurement of the binding potential of the dopamine transporter radioligand ^123I-ioflupane in mouse brain in the awake and anesthetized (isoflurane) states. Stress induced by imaging in the awake state was assessed through measurement of plasma corticosterone levels. Results: AwakeSPECT provided high-resolution bone images reminiscent of those obtained from CT. The binding potential of ^123I-ioflupane in the awake state was on the order of 50% of that obtained with the animal under anesthesia, consistent with previous studies in nonhuman primates. Levels of stress induced were on the order of those seen in other behavioral tasks and imaging studies of awake animals. Conclusion: These results demonstrate the feasibility of SPECT molecular brain imaging of mice in the conscious, unrestrained state and demonstrate the effects of isoflurane anesthesia on radiotracer uptake.

  14. Aspirin-triggered resolvin D1 down-regulates inflammatory responses and protects against endotoxin-induced acute kidney injury

    SciTech Connect (OSTI)

    Chen, Jiao; Shetty, Sreerama; Zhang, Ping; Gao, Rong; Hu, Yuxin; Wang, Shuxia; Li, Zhenyu; Fu, Jian

    2014-06-01

    The presence of endotoxin in blood can lead to acute kidney injury (AKI) and septic shock. Resolvins, the endogenous lipid mediators derived from docosahexaenoic acid, have been reported to exhibit potent anti-inflammatory action. Using a mouse model of lipopolysaccharide (LPS)-induced AKI, we investigated the effects of aspirin-triggered resolvin D1 (AT-RvD1) on inflammatory kidney injury. Administration of AT-RvD1 1 h after LPS challenge protected the mice from kidney injury as indicated by the measurements of blood urea nitrogen, serum creatinine, and morphological alterations associated with tubular damage. The protective effects were evidenced by decreased neutrophil infiltration in the kidney indicating reduction in inflammation. AT-RvD1 treatment restored kidney cell junction protein claudin-4 expression, which was otherwise reduced after LPS challenge. AT-RvD1 treatment inhibited endotoxin-induced NF-κB activation and suppressed LPS-induced ICAM-1 and VCAM-1 expression in the kidney. Moreover, AT-RvD1 treatment markedly decreased LPS-induced IL-6 level in the kidney and blocked IL-6-mediated signaling including STAT3 and ERK phosphorylation. Our findings demonstrate that AT-RvD1 is a potent anti-inflammatory mediator in LPS-induced kidney injury, and AT-RvD1 has therapeutic potential against AKI during endotoxemia.

  15. Activation of farnesoid X receptor attenuates hepatic injury in a murine model of alcoholic liver disease

    SciTech Connect (OSTI)

    Wu, Weibin; Institutes of Biomedical Science, Fudan University, Shanghai 200032 ; Zhu, Bo; Peng, Xiaomin; Zhou, Meiling; Jia, Dongwei; Gu, Jianxin; Institutes of Biomedical Science, Fudan University, Shanghai 200032

    2014-01-03

    Highlights: FXR activity was impaired by chronic ethanol ingestion in a murine model of ALD. Activation of FXR attenuated alcohol-induced liver injury and steatosis. Activation of FXR attenuated cholestasis and oxidative stress in mouse liver. -- Abstract: Alcoholic liver disease (ALD) is a common cause of advanced liver disease, and considered as a major risk factor of morbidity and mortality worldwide. Hepatic cholestasis is a pathophysiological feature observed in all stages of ALD. The farnesoid X receptor (FXR) is a member of the nuclear hormone receptor superfamily, and plays an essential role in the regulation of bile acid, lipid and glucose homeostasis. However, the role of FXR in the pathogenesis and progression of ALD remains largely unknown. Mice were fed Lieber-DeCarli ethanol diet or an isocaloric control diet. We used a specific agonist of FXR WAY-362450 to study the effect of pharmacological activation of FXR in alcoholic liver disease. In this study, we demonstrated that FXR activity was impaired by chronic ethanol ingestion in a murine model of ALD. Activation of FXR by specific agonist WAY-362450 protected mice from the development of ALD. We also found that WAY-362450 treatment rescued FXR activity, suppressed ethanol-induced Cyp2e1 up-regulation and attenuated oxidative stress in liver. Our results highlight a key role of FXR in the modulation of ALD development, and propose specific FXR agonists for the treatment of ALD patients.

  16. Structure of Human Ferritin L Chain

    SciTech Connect (OSTI)

    Wang,Z.; Li, C.; Ellenburg, M.; Soistman, E.; Ruble, J.; Wright, B.; Ho, J.; Carter, D.

    2006-01-01

    Ferritin is the major iron-storage protein present in all cells. It generally contains 24 subunits, with different ratios of heavy chain (H) to light chain (L), in the shape of a hollow sphere hosting up to 4500 ferric Fe atoms inside. H-rich ferritins catalyze the oxidation of iron(II), while L-rich ferritins promote the nucleation and storage of iron(III). Several X-ray structures have been determined, including those of L-chain ferritins from horse spleen (HoSF), recombinant L-chain ferritins from horse (HoLF), mouse (MoLF) and bullfrog (BfLF) as well as recombinant human H-chain ferritin (HuHF). Here, structures have been determined of two crystal forms of recombinant human L-chain ferritin (HuLF) obtained from native and perdeuterated proteins. The structures show a cluster of acidic residues at the ferrihydrite nucleation site and at the iron channel along the threefold axis. An ordered Cd{sup 2+} structure is observed within the iron channel, offering further insight into the route and mechanism of iron transport into the capsid. The loop between helices D and E, which is disordered in many other L-chain structures, is clearly visible in these two structures. The crystals generated from perdeuterated HuLF will be used for neutron diffraction studies.

  17. Transcriptional and metabolic flux profiling of triadimefon effects on cultured hepatocytes

    SciTech Connect (OSTI)

    Iyer, Vidya V.; Ovacik, Meric A.; Androulakis, Ioannis P.; Roth, Charles M.; Ierapetritou, Marianthi G.

    2010-11-01

    Conazoles are a class of azole fungicides used to prevent fungal growth in agriculture, for treatment of fungal infections, and are found to be tumorigenic in rats and/or mice. In this study, cultured primary rat hepatocytes were treated to two different concentrations (0.3 and 0.15 mM) of triadimefon, which is a tumorigenic conazole in rat and mouse liver, on a temporal basis with daily media change. Following treatment, cells were harvested for microarray data ranging from 6 to 72 h. Supernatant was collected daily for three days, and the concentrations of various metabolites in the media and supernatant were quantified. Gene expression changes were most significant following exposure to 0.3 mM triadimefon and were characterized mainly by metabolic pathways related to carbohydrate, lipid and amino acid metabolism. Correspondingly, metabolic network flexibility analysis demonstrated a switch from fatty acid synthesis to fatty acid oxidation in cells exposed to triadimefon. It is likely that fatty acid oxidation is active in order to supply energy required for triadimefon detoxification. In 0.15 mM triadimefon treatment, the hepatocytes are able to detoxify the relatively low concentration of triadimefon with less pronounced changes in hepatic metabolism.

  18. Low Dose Radiation Response Curves, Networks and Pathways in Human Lymphoblastoid Cells Exposed from 1 to 10 cGy of Acute Gamma Radiation

    SciTech Connect (OSTI)

    Wyrobek, A. J.; Manohar, C. F.; Nelson, D. O.; Furtado, M. R.; Bhattacharya, M. S.; Marchetti, F.; Coleman, M.A.

    2011-04-18

    We investigated the low dose dependency of the transcriptional response of human cells to characterize the shape and biological functions associated with the dose response curve and to identify common and conserved functions of low dose expressed genes across cells and tissues. Human lymphoblastoid (HL) cells from two unrelated individuals were exposed to graded doses of radiation spanning the range of 1-10 cGy were analyzed by transcriptome profiling, qPCR and bioinformatics, in comparison to sham irradiated samples. A set of {approx}80 genes showed consistent responses in both cell lines; these genes were associated with homeostasis mechanisms (e.g., membrane signaling, molecule transport), subcellular locations (e.g., Golgi, and endoplasmic reticulum), and involved diverse signal transduction pathways. The majority of radiation-modulated genes had plateau-like responses across 1-10 cGy, some with suggestive evidence that transcription was modulated at doses below 1 cGy. MYC, FOS and TP53 were the major network nodes of the low-dose response in HL cells. Comparison our low dose expression findings in HL cells with those of prior studies in mouse brain after whole body exposure, in human keratinocyte cultures, and in endothelial cells cultures, indicates that certain components of the low dose radiation response are broadly conserved across cell types and tissues, independent of proliferation status.

  19. Nuclear substructure reorganization during late stageerythropoiesis is selective and does not involve caspase cleavage ofmajor nuclear substructural proteins

    SciTech Connect (OSTI)

    Krauss, Sharon Wald; Lo, Annie J.; Short, Sarah A.; Koury, MarkJ.; Mohandas, Narla; Chasis, Joel Anne

    2005-04-06

    Enucleation, a rare feature of mammalian differentiation, occurs in three cell types: erythroblasts, lens epithelium and keratinocytes. Previous investigations suggest that caspase activation functions in lens epithelial and keratinocyte enucleation, as well as in early erythropoiesis encompassing BFU-E differentiation to proerythroblast. To determine whether caspase activation contributes to later erythropoiesis and whether nuclear substructures other than chromatin reorganize, we analyzed distributions of nuclear subcompartment proteins and assayed for caspase-induced cleavage of subcompartmental target proteins in mouse erythroblasts. We found that patterns of lamin B in the filamentous network interacting with both the nuclear envelope and DNA, nuclear matrix protein NuMA, and splicing factors Sm and SC35 persisted during nuclear condensation, consistent with effective transcription of genes expressed late in differentiation. Thus nuclear reorganization prior to enucleation is selective, allowing maintenance of critical transcriptional processes independent of extensive chromosomal reorganization. Consistent with these data, we found no evidence for caspase-induced cleavage of major nuclear subcompartment proteins during late erythropoiesis, in contrast to what has been observed in early erythropoiesis and in lens epithelial and keratinocyte differentiation. These findings imply that nuclear condensation and extrusion during terminal erythroid differentiation involve novel mechanisms that do not entail major activation of apoptotic machinery.

  20. Enhanced human papillomavirus type 8 oncogene expression levels are crucial for skin tumorigenesis in transgenic mice

    SciTech Connect (OSTI)

    Hufbauer, M.; Lazic, D.; Akguel, B.; Brandsma, J.L.; Pfister, H.; Weissenborn, S.J.

    2010-08-01

    Human papillomavirus 8 (HPV8) is involved in skin cancer development in epidermodysplasia verruciformis patients. Transgenic mice expressing HPV8 early genes (HPV8-CER) developed papillomas, dysplasias and squamous cell carcinomas. UVA/B-irradiation and mechanical wounding of HPV8-CER mouse skin led to prompt papilloma induction in about 3 weeks. The aim of this study was to analyze the kinetics and level of transgene expression in response to skin irritations. Transgene expression was already enhanced 1 to 2 days after UVA/B-irradiation or tape-stripping and maintained during papilloma development. The enhanced transgene expression could be assigned to UVB and not to UVA. Papilloma development was thus always paralleled by an increased transgene expression irrespective of the type of skin irritation. A knock-down of E6 mRNA by tattooing HPV8-E6-specific siRNA led to a delay and a lower incidence of papilloma development. This indicates that the early increase of viral oncogene expression is crucial for induction of papillomatosis.

  1. A miniature mimic of host defense peptides with systemic antibacterial efficacy

    SciTech Connect (OSTI)

    Sarig, Hadar; Livne, Liran; Held-Kuznetsov, Victoria; Zaknoon, Fadia; Ivankin, Andrey; Gidalevitz, David; Mor, Amram

    2010-08-23

    Oligomers of acylated lysines (OAKs) are synthetic mimics of host defense peptides (HDPs) with promising antimicrobial properties. Here we challenged the OAK concept for its ability to generate both systemically efficient and economically viable lead compounds for fighting multidrug-resistant bacteria. We describe the design and characterization of a miniature OAK composed of only 3 lysyls and 2 acyls (designated C{sub 12({omega}7)}K-{beta}{sub 12}) that preferentially targets gram-positive species by a bacteriostatic mode of action. To gain insight into the mechanism of action, we examined the interaction of OAK with various potential targets, including phospholipid bilayers, using surface plasmon resonance, and Langmuir monolayers, using insertion assays, epifluorescence microscopy, and grazing incidence X-ray diffraction, in a complementary manner. Collectively, the data support the notion that C{sub 12({omega}7)}K-{beta}{sub 12} damages the plasma-membrane architecture similarly to HDPs, that is, following a near-classic 2-step interaction including high-affinity electrostatic adhesion and a subsequent shallow insertion that was limited to the phospholipid head group region. Notably, preliminary acute toxicity and efficacy studies performed with mouse models of infection have consolidated the potential of OAK for treating bacterial infections, including systemic treatments of methicillin-resistant Staphylococcus aureus. Such simple yet robust chemicals might be useful for various antibacterial applications while circumventing potential adverse effects associated with cytolytic compounds.

  2. Understanding dynamic changes in live cell adhesion with neutron reflectometry

    SciTech Connect (OSTI)

    Junghans, Ann; Waltman, Mary Jo; Smith, Hillary L.; Pocivavsek, Luka; Zebda, Noureddine; Birukov, Konstantin; Viapiano, Mariano; Majewski, Jaroslaw

    2014-12-10

    In this study, neutron reflectometry (NR) was used to examine various live cells' adhesion to quartz substrates under different environmental conditions, including flow stress. To the best of our knowledge, these measurements represent the first successful visualization and quantization of the interface between live cells and a substrate with sub-nanometer resolution. In our first experiments, we examined live mouse fibroblast cells as opposed to past experiments using supported lipids, proteins, or peptide layers with no associated cells. We continued the NR studies of cell adhesion by investigating endothelial monolayers and glioblastoma cells under dynamic flow conditions. We demonstrated that neutron reflectometry is a powerful tool to study the strength of cellular layer adhesion in living tissues, which is a key factor in understanding the physiology of cell interactions and conditions leading to abnormal or disease circumstances. Continuative measurements, such as investigating changes in tumor cell — surface contact of various glioblastomas, could impact advancements in tumor treatments. In principle, this can help us to identify changes that correlate with tumor invasiveness. Pursuit of these studies can have significant medical impact on the understanding of complex biological problems and their effective treatment, e.g. for the development of targeted anti-invasive therapies.

  3. Effective delivery of recombinant proteins to rod photoreceptors via lipid nanovesicles

    SciTech Connect (OSTI)

    Asteriti, Sabrina; Dal Cortivo, Giuditta; Pontelli, Valeria; Cangiano, Lorenzo; Buffelli, Mario; Dell’Orco, Daniele

    2015-06-12

    The potential of liposomes to deliver functional proteins in retinal photoreceptors and modulate their physiological response was investigated by two experimental approaches. First, we treated isolated mouse retinas with liposomes encapsulating either recoverin, an important endogenous protein operating in visual phototransduction, or antibodies against recoverin. We then intravitrally injected in vivo liposomes encapsulating either rhodamin B or recoverin and we investigated the distribution in retina sections by confocal microscopy. The content of liposomes was found to be released in higher amount in the photoreceptor layer than in the other regions of the retina and the functional effects of the release were in line with the current model of phototransduction. Our study sets the basis for quantitative investigations aimed at assessing the potential of intraocular protein delivery via biocompatible nanovesicles, with promising implications for the treatment of retinal diseases affecting the photoreceptor layer. - Highlights: • Recombinant proteins encapsulated in nano-sized liposomes injected intravitreally reach retinal photoreceptors. • The phototransduction cascade in rods is modulated by the liposome content. • Mathematical modeling predicts the alteration of the photoresponses following liposome fusion.

  4. Assessment of the efficacy of laser hyperthermia and nanoparticle-enhanced therapies by heat shock protein analysis

    SciTech Connect (OSTI)

    Tang, Fei; Zhang, Ye; Zhang, Juan; Liu, Ran; Guo, Junwei

    2014-03-15

    Tumor thermotherapy is a method of cancer treatment wherein cancer cells are killed by exposing the body tissues to high temperatures. Successful clinical implementation of this method requires a clear understanding and assessment of the changes of the tumor area after the therapy. In this study, we evaluated the effect of near-infrared laser tumor thermotherapy at the molecular, cellular, and physical levels. We used single-walled carbon nanotubes (SWNTs) in combination with this thermotherapy. We established a mouse model for breast cancer and randomly divided the mice into four groups: mice with SWNT-assisted thermotherapy; mice heat treated without SWNT; mice injected with SWNTs without thermotherapy; and a control group. Tumors were irradiated using a near-infrared laser with their surface temperature remaining at approximately 45 C. We monitored the tumor body growth trend closely by daily physical measurements, immunohistochemical staining, and H and E (hematoxylin-eosin) staining by stage. Our results showed that infrared laser hyperthermia had a significant inhibitory effect on the transplanted breast tumor, with an inhibition rate of 53.09%, and also significantly reduced the expression of the heat shock protein Hsp70. Furthermore, we have found that protein analysis and histological analysis can be used to assess therapeutic effects effectively, presenting broad application prospects for determining the effect of different treatments on tumors. Finally, we discuss the effects of SWNT-assisted near-infrared laser tumor thermotherapy on tumor growth at the molecular, cellular, and physical levels.

  5. Centrifugal microfluidic platform for ultrasensitive detection of botulinum toxin

    SciTech Connect (OSTI)

    Koh, Chung -Yan; Schaff, Ulrich Y.; Piccini, Matthew E.; Stanker, Larry H.; Cheng, Luisa W.; Ravichandran, Easwaran; Singh, Bal -Ram; Sommer, Greg J.; Singh, Anup K.

    2014-12-18

    In this study, we present an innovative centrifugal microfluidic immunoassay platform (SpinDx) to address the urgent biodefense and public health need for ultrasensitive point-of-care/incident detection of botulinum toxin. The simple, sample-to-answer centrifugal microfluidic immunoassay approach is based on binding of toxins to antibody-laden capture particles followed by sedimentation of the particles through a density-media in a microfluidic disk and quantification by laser-induced fluorescence. A blind, head-to-head comparison study of SpinDx versus the gold-standard mouse bioassay demonstrates 100-fold improvement in sensitivity (limit of detection = 0.09 pg/mL), while achieving total sample-to-answer time of <30 min with 2-μL required volume of the unprocessed sample. We further demonstrate quantification of botulinum toxin in both exogeneous (human blood and serum spiked with toxins) and endogeneous (serum from mice intoxicated via oral, intranasal, and intravenous routes) samples. SpinDx can analyze, without any sample preparation, multiple sample types including whole blood, serum, and food. It is readily expandable to additional analytes as the assay reagents (i.e., the capture beads and detection antibodies) are disconnected from the disk architecture and the reader, facilitating rapid development of new assays. SpinDx can also serve as a general-purpose immunoassay platform applicable to diagnosis of other conditions and diseases.

  6. Numerical calculation of protein-ligand binding rates through solution of the Smoluchowski equation using smoothed particle hydrodynamics

    SciTech Connect (OSTI)

    Pan, Wenxiao; Daily, Michael; Baker, Nathan A.

    2015-05-07

    Background: The calculation of diffusion-controlled ligand binding rates is important for understanding enzyme mechanisms as well as designing enzyme inhibitors. Methods: We demonstrate the accuracy and effectiveness of a Lagrangian particle-based method, smoothed particle hydrodynamics (SPH), to study diffusion in biomolecular systems by numerically solving the time-dependent Smoluchowski equation for continuum diffusion. Unlike previous studies, a reactive Robin boundary condition (BC), rather than the absolute absorbing (Dirichlet) BC, is considered on the reactive boundaries. This new BC treatment allows for the analysis of enzymes with imperfect reaction rates. Results: The numerical method is first verified in simple systems and then applied to the calculation of ligand binding to a mouse acetylcholinesterase (mAChE) monomer. Rates for inhibitor binding to mAChE are calculated at various ionic strengths and compared with experiment and other numerical methods. We find that imposition of the Robin BC improves agreement between calculated and experimental reaction rates. Conclusions: Although this initial application focuses on a single monomer system, our new method provides a framework to explore broader applications of SPH in larger-scale biomolecular complexes by taking advantage of its Lagrangian particle-based nature.

  7. Identification of stem cells from human umbilical cord blood with embryonic and hematopoietic characteristics

    SciTech Connect (OSTI)

    Zhao Yong . E-mail: yongzhao@uic.edu; Wang Honglan; Mazzone, Theodore

    2006-08-01

    We identified stem cells from the umbilical cord blood, designated cord blood-stem cells (CB-SC). CB-SC displayed important embryonic stem (ES) cell characteristics including expression of ES-cell-specific molecular markers including transcription factors OCT-4 and Nanog, along with stage-specific embryonic antigen (SSEA)-3 and SSEA-4. CB-SC also expressed hematopoietic cell antigens including CD9, CD45 and CD117, but were negative for CD34. CB-SC displayed very low immunogenicity as indicated by expression of a very low level of major histocompatibility complex (MHC) antigens and failure to stimulate the proliferation of allogeneic lymphocytes. CB-SC could give rise to cells with endothelial-like and neuronal-like characteristics in vitro, as demonstrated by expression of lineage-associated markers. Notably, CB-SC could be stimulated to differentiate into functional insulin-producing cells in vivo and eliminated hyperglycemia after transplantation into a streptozotocin-induced diabetic mouse model. These findings may have significant potential to advance stem-cell-based therapeutics.

  8. A Semiautomated Framework for Integrating Expert Knowledge into Disease Marker Identification

    SciTech Connect (OSTI)

    Wang, Jing; Webb-Robertson, Bobbie-Jo M.; Matzke, Melissa M.; Varnum, Susan M.; Brown, Joseph N.; Riensche, Roderick M.; Adkins, Joshua N.; Jacobs, Jon M.; Hoidal, John R.; Scholand, Mary Beth; Pounds, Joel G.; Blackburn, Michael R.; Rodland, Karin D.; McDermott, Jason E.

    2013-10-01

    Background. The availability of large complex data sets generated by high throughput technologies has enabled the recent proliferation of disease biomarker studies. However, a recurring problem in deriving biological information from large data sets is how to best incorporate expert knowledge into the biomarker selection process. Objective. To develop a generalizable framework that can incorporate expert knowledge into data-driven processes in a semiautomated way while providing a metric for optimization in a biomarker selection scheme. Methods. The framework was implemented as a pipeline consisting of five components for the identification of signatures from integrated clustering (ISIC). Expert knowledge was integrated into the biomarker identification process using the combination of two distinct approaches; a distance-based clustering approach and an expert knowledge-driven functional selection. Results. The utility of the developed framework ISIC was demonstrated on proteomics data from a study of chronic obstructive pulmonary disease (COPD). Biomarker candidates were identified in a mouse model using ISIC and validated in a study of a human cohort. Conclusions. Expert knowledge can be introduced into a biomarker discovery process in different ways to enhance the robustness of selected marker candidates. Developing strategies for extracting orthogonal and robust features from large data sets increases the chances of success in biomarker identification.

  9. Catalog of gene expression in adult neural stem cells and their in vivo microenvironment

    SciTech Connect (OSTI)

    Williams, Cecilia; Wirta, Valtteri; Meletis, Konstantinos; Wikstroem, Lilian; Carlsson, Leif; Frisen, Jonas; Lundeberg, Joakim . E-mail: joakim.lundeberg@biotech.kth.se

    2006-06-10

    Stem cells generally reside in a stem cell microenvironment, where cues for self-renewal and differentiation are present. However, the genetic program underlying stem cell proliferation and multipotency is poorly understood. Transcriptome analysis of stem cells and their in vivo microenvironment is one way of uncovering the unique stemness properties and provides a framework for the elucidation of stem cell function. Here, we characterize the gene expression profile of the in vivo neural stem cell microenvironment in the lateral ventricle wall of adult mouse brain and of in vitro proliferating neural stem cells. We have also analyzed an Lhx2-expressing hematopoietic-stem-cell-like cell line in order to define the transcriptome of a well-characterized and pure cell population with stem cell characteristics. We report the generation, assembly and annotation of 50,792 high-quality 5'-end expressed sequence tag sequences. We further describe a shared expression of 1065 transcripts by all three stem cell libraries and a large overlap with previously published gene expression signatures for neural stem/progenitor cells and other multipotent stem cells. The sequences and cDNA clones obtained within this framework provide a comprehensive resource for the analysis of genes in adult stem cells that can accelerate future stem cell research.

  10. Centrifugal microfluidic platform for ultrasensitive detection of botulinum toxin

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Koh, Chung -Yan; Schaff, Ulrich Y.; Sandstone Diagnostics, Livermore, CA; Piccini, Matthew E.; Cepheid, Sunnyvale, CA; Stanker, Larry H.; Cheng, Luisa W.; Ravichandran, Easwaran; Singh, Bal -Ram; Sommer, Greg J.; et al

    2014-12-18

    In this study, we present an innovative centrifugal microfluidic immunoassay platform (SpinDx) to address the urgent biodefense and public health need for ultrasensitive point-of-care/incident detection of botulinum toxin. The simple, sample-to-answer centrifugal microfluidic immunoassay approach is based on binding of toxins to antibody-laden capture particles followed by sedimentation of the particles through a density-media in a microfluidic disk and quantification by laser-induced fluorescence. A blind, head-to-head comparison study of SpinDx versus the gold-standard mouse bioassay demonstrates 100-fold improvement in sensitivity (limit of detection = 0.09 pg/mL), while achieving total sample-to-answer time of <30 min with 2-μL required volume of themore » unprocessed sample. We further demonstrate quantification of botulinum toxin in both exogeneous (human blood and serum spiked with toxins) and endogeneous (serum from mice intoxicated via oral, intranasal, and intravenous routes) samples. SpinDx can analyze, without any sample preparation, multiple sample types including whole blood, serum, and food. It is readily expandable to additional analytes as the assay reagents (i.e., the capture beads and detection antibodies) are disconnected from the disk architecture and the reader, facilitating rapid development of new assays. SpinDx can also serve as a general-purpose immunoassay platform applicable to diagnosis of other conditions and diseases.« less

  11. Cul4A overexpression associated with Gli1 expression in malignant pleural mesothelioma

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Yang, Yi -Lin; Ni, Jian; Hsu, Ping -Chih; Mao, Jian -Hua; Hsieh, David; Xu, Angela; Chan, Geraldine; Au, Alfred; Xu, Zhidong; Jablons, David M.; et al

    2015-07-27

    Malignant pleural mesothelioma (mesothelioma) is a highly aggressive cancer without an effective treatment. Cul4A, a scaffold protein that recruits substrates for degradation, is amplified in several human cancers, including mesothelioma. We have recently shown that Cul4A plays an oncogenic role in vitro and in a mouse model. In this study, we analysed clinical mesothelioma tumours and found moderate to strong expression of Cul4A in 70.9% (51/72) of these tumours, as shown by immunohistochemistry. In 72.2% mesothelioma tumours with increased Cul4A copy number identified by fluorescence in situ hybridization analysis, Cul4A protein expression was moderate to strong. Similarly, Cul4A was overexpressedmore » and Cul4A copy number was increased in human mesothelioma cell lines. Because Gli1 is highly expressed in human mesothelioma cells, we compared Cul4A and Gli1 expression in mesothelioma tumours and found their expression associated (P < 0.05, chi-square). In mesothelioma cell lines, inhibiting Cul4A by siRNA decreased Gli1 expression, suggesting that Gli1 expression is, at least in part, regulated by Cul4A in mesothelioma cells. Our results suggest a linkage between Cul4A and Gli1 expression in human mesothelioma.« less

  12. Molecular cytogenetic characterization of a human thyroid cancercell line

    SciTech Connect (OSTI)

    Weier, Heinz-Ulrich G.; Tuton, Tiffany B.; Ito, Yuko; Chu, LisaW.; Lu, Chung-Mei; Baumgartner, Adolf; Zitzelsberger, Horst F.; Weier,Jingly F.

    2006-01-04

    The incidence of papillary thyroid carcinoma (PTC) increases significantly after exposure of the head and neck region to ionizing radiation, yet we know neither the steps involved in malignant transformation of thyroid epithelium nor the specific carcinogenic mode of action of radiation. Such increased tumor frequency became most evident in children after the 1986 nuclear accident in Chernobyl, Ukraine. In the twelve years following the accident, the average incidence of childhood PTCs (chPTC) increased over one hundred-fold compared to the rate of about 1 tumor incidence per 10{sup 6} children per year prior to 1986. To study the etiology of radiation-induced thyroid cancer, we formed an international consortium to investigate chromosomal changes and altered gene expression in cases of post-Chernobyl chPTC. Our approach is based on karyotyping of primary cultures established from chPTC specimens, establishment of cell lines and studies of genotype-phenotype relationships through high resolution chromosome analysis, DNA/cDNA micro-array studies, and mouse xenografts that test for tumorigenicity. Here, we report the application of fluorescence in situ hybridization (FISH)-based techniques for the molecular cytogenetic characterization of a highly tumorigenic chPTC cell line, S48TK, and its subclones. Using chromosome 9 rearrangements as an example, we describe a new approach termed ''BAC-FISH'' to rapidly delineate chromosomal breakpoints, an important step towards a better understanding of the formation of translocations and their functional consequences.

  13. Inhibition of K+ permeability diminishes alpha 2-adrenoceptor mediated effects on norepinephrine release

    SciTech Connect (OSTI)

    Zimanyi, I.; Folly, G.; Vizi, E.S.

    1988-05-01

    The effect of two different potassium channel blockers, 4-aminopyridine (4-AP) and quinine, on the alpha 2-adrenoceptor mediated modulation of norepinephrine (NE) release was investigated. Pairs of mouse vasa deferentia were loaded with /sup 3/H-norepinephrine (/sup 3/H-NE), superfused continuously, and stimulated electrically. 4-AP (5.3 x 10(-4) M), and quinine (10(-5) M) enhanced the stimulation-evoked release of tritium significantly. The electrically induced release of radioactivity was reduced by alpha 2-adrenoceptor agonists (1-NE and xylazine) and enhanced by the alpha 2-adrenoceptor antagonist yohimbine. Both effects were affected markedly by 4-AP or quinine: the depressant action of 1-NA and xylazine was partially antagonized and the facilitatory effect of yohimbine was completely abolished during the blockade of the potassium channels. It is suggested that the blockade of the potassium permeability counteracts negative feedback modulation; therefore, it seems likely that the stimulation of alpha 2-adrenoceptors leads to an enhanced potassium permeability and hyperpolarization of varicose axon terminals.

  14. High-resolution imaging of selenium in kidneys: a localized selenium pool associated with glutathione peroxidase 3

    SciTech Connect (OSTI)

    Malinouski, M.; Kehr, S.; Finney, L.; Vogt, S.; Carlson, B.A.; Seravalli, J.; Jin, R.; Handy, D.E.; Park, T.J.; Loscalzo, J.; Hatfield, D.L.; Gladyshev, V.N.

    2012-04-17

    Recent advances in quantitative methods and sensitive imaging techniques of trace elements provide opportunities to uncover and explain their biological roles. In particular, the distribution of selenium in tissues and cells under both physiological and pathological conditions remains unknown. In this work, we applied high-resolution synchrotron X-ray fluorescence microscopy (XFM) to map selenium distribution in mouse liver and kidney. Liver showed a uniform selenium distribution that was dependent on selenocysteine tRNA{sup [Ser]Sec} and dietary selenium. In contrast, kidney selenium had both uniformly distributed and highly localized components, the latter visualized as thin circular structures surrounding proximal tubules. Other parts of the kidney, such as glomeruli and distal tubules, only manifested the uniformly distributed selenium pattern that co-localized with sulfur. We found that proximal tubule selenium localized to the basement membrane. It was preserved in Selenoprotein P knockout mice, but was completely eliminated in glutathione peroxidase 3 (GPx3) knockout mice, indicating that this selenium represented GPx3. We further imaged kidneys of another model organism, the naked mole rat, which showed a diminished uniformly distributed selenium pool, but preserved the circular proximal tubule signal. We applied XFM to image selenium in mammalian tissues and identified a highly localized pool of this trace element at the basement membrane of kidneys that was associated with GPx3. XFM allowed us to define and explain the tissue topography of selenium in mammalian kidneys at submicron resolution.

  15. The Near Naked Hairless (HrN) Mutation Disrupts Hair Formation but is not Due to a Mutation in the Hairless Coding Region

    SciTech Connect (OSTI)

    Liu, Yutao; Das, Suchita; Olszewski, Robert Edward; Culiat, Cymbeline T; Carpenter, D A; Sundberg, John P; Soteropoulos, Patricia; Liu, Xiaochen; Doktycz, Mitchel John; Michaud III, Edward J; Voy, Brynn H

    2007-01-01

    Near naked hairless (HrN) is a semi-dominant mutation that arose spontaneously and was suggested by allelism testing to be an allele of mouse Hairless (Hr). HrN mice differ from other Hr mutants in that hair loss appears as the postnatal coat begins to emerge, as opposed to failure to initiate the first postnatal hair cycle, and that the mutation displays semi-dominant inheritance. We sequenced the Hr cDNA in HrN/HrN mice and characterized the pathological and molecular phenotypes to identify the basis for hair loss in this model. HrN/HrN mice exhibit dystrophic hairs that are unable to consistently emerge from the hair follicle, while HrN/+ mice display a sparse coat of hair and a milder degree of follicular dystrophy than their homozygous littermates. DNA microarray analysis of cutaneous gene expression demonstrates that numerous genes are downregulated in HrN/HrN mice, primarily genes important for hair structure. By contrast, Hr expression is significantly increased. Sequencing the Hr coding region, intron-exon boundaries, 5'- and 3'- UTR and immediate upstream region did not reveal the underlying mutation. Therefore HrN does not appear to be an allele of Hr but may result from a mutation in a closely linked gene or from a regulatory mutation in Hr.

  16. A possible role for the canonical Wnt pathway in endocrine cell development in chicks

    SciTech Connect (OSTI)

    Pedersen, Anna Hauntoft; Heller, R. Scott . E-mail: shll@hagedorn.dk

    2005-08-05

    Wnt signalling is involved in many developmental processes such as proliferation, differentiation, cell fate decisions, and morphogenesis. However, little is known about Wnt signalling during pancreas development. Multiple Wnt ligands and Frizzled receptors are expressed in the embryonic mouse pancreas, the surrounding mesenchyme, and have also been detected in the chicken endoderm during development. The aim of this study was to investigate the role of canonical Wnt signalling on endocrine cell development by use of the in ovo electroporation of the chicken endoderm. Overexpression with a constitutive active form of {beta}-catenin in combination with Ngn3 resulted in reduced numbers of glucagon cells. dnLEF-1 or naked-1 did not alter endocrine cell differentiation when co-expressed with Ngn3, but dnLEF-1 appeared to have some potential for inhibiting delamination of Ngn3 cells. In addition, neuronal {beta}-III-tubulin, which had previously been considered a specific marker for neuronal cells, was observed in the pancreas and was upregulated in the electroporated Ngn3 cells and thus may be a new endocrine marker in the chicken.

  17. Identification of genetic loci that control mammary tumor susceptibility through the host microenvironment

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Zhang, Pengju; Lo, Alvin; Huang, Yurong; Huang, Ge; Liang, Guozhou; Mott, Joni; Karpen, Gary H.; Blakely, Eleanor A.; Bissell, Mina J.; Barcellos-Hoff, Mary Helen; et al

    2015-03-09

    The interplay between host genetics, tumor microenvironment and environmental exposure in cancer susceptibility remains poorly understood. Here we assessed the genetic control of stromal mediation of mammary tumor susceptibility to low dose ionizing radiation (LDIR) using backcrossed F1 into BALB/c (F1Bx) between cancer susceptible (BALB/c) and resistant (SPRET/EiJ) mouse strains. Tumor formation was evaluated after transplantation of non-irradiated Trp53-/- BALB/c mammary gland fragments into cleared fat pads of F1Bx hosts. Genome-wide linkage analysis revealed 2 genetic loci that constitute the baseline susceptibility via host microenvironment. However, once challenged with LDIR, we discovered 13 additional loci that were enriched for genesmore » involved in cytokines, including TGFβ1 signaling. Surprisingly, LDIR-treated F1Bx cohort significantly reduced incidence of mammary tumors from Trp53-/- fragments as well as prolonged tumor latency, compared to sham-treated controls. We demonstrated further that plasma levels of specific cytokines were significantly correlated with tumor latency. Using an ex vivo 3-D assay, we confirmed TGFβ1 as a strong candidate for reduced mammary invasion in SPRET/EiJ, which could explain resistance of this strain to mammary cancer risk following LDIR. Our results open possible new avenues to understand mechanisms of genes operating via the stroma that affect cancer risk from external environmental exposures.« less

  18. The Adipose Renin-Angiotensin System Modulates Systemic Markers of Insulin Sensitivity and Activates the Intrarenal Renin-Angiotensin System

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Kim, Suyeon; Soltani-Bejnood, Morvarid; Quignard-Boulange, Annie; Massiera, Florence; Teboul, Michele; Ailhaud, Gerard; Kim, Jung Han; Moustaid-Moussa, Naima; Voy, Brynn H.

    2006-01-01

    Background . The adipose tissue renin-angiotensin system (RAS) contributes to regulation of fat mass and may also impact systemic functions such as blood pressure and metabolism. Methods and results . A panel of mouse models including mice lacking angiotensinogen, Agt ( Agt -KO), mice expressing Agt solely in adipose tissue (aP2- Agt/Agt -KO), and mice overexpressing Agt in adipose tissue (aP2- Agt ) was studied. Total body weight, epididymal fat pad weight, and circulating levels of leptin, insulin, and resistin were significantly decreased in Agt -KO mice, while plasma adiponectin levels were increased. aP2- Agt mice exhibited increased adipositymore » and plasma leptin and insulin levels compared to wild type (WT) controls. Angiotensinogen and type I Ang II receptor protein levels were also elevated in kidney of aP2- Agt mice. Conclusion . These findings demonstrate that alterations in adipose RAS activity significantly impact both local and systemic physiology in a way that may contribute to the detrimental health effects of obesity.« less

  19. Polymorphism of the MHC class II Eb gene determines the protection against collagen-induced arthritis

    SciTech Connect (OSTI)

    Gonzalez-Gay, M.A.; Zanelli, E.; Krco, C.J.

    1995-05-01

    Collagen-induced arthritis (CIA) is an animal model of auto immune polyarthritis, sharing similarities with rheumatoid arthritis (RA). Paradoxally, susceptibility to mouse CIA is controlled by the H2A loci (DQ homologous) while RA is linked to HLA.DR genes (H2E homologous). We recently showed that the E{beta}{sup d} molecule prevents CIA development in susceptible H2{sup q} mice. We addressed the question of whether H2Eb polymorphism will influence CIA incidence as HLA.DRB1 polymorphism does in RA. In F{sub 1} mice, only H2Eb{sup d} and H2Eb{sup s} molecules showed protection. Using recombinant B10.RDD (Eb{sup d/b}) mice, we found that CIA protection was mediated by the first domain of the E{beta}{sup d} molecule. Using peptides covering the third hypervariable region of the E{beta} chain, we found a perfect correlation between presentation of E{beta} peptides by the H2A{sup q} molecule and protection on CIA. Therefore, the mechanism by which H2Eb protects against CIA seems to rely on the affinity of E{beta} peptides for the H2A{sup q} molecule. 35 refs., 2 figs., 3 tabs.

  20. Relationship of DNA repair processes to mutagenesis and carcinogenesis in mammalian cells. Progress report, August 1, 1977-October 31, 1980

    SciTech Connect (OSTI)

    Evans, H.H.

    1980-10-01

    The objective of this research is to determine the role of DNA repair in mutagenesis and carcinogenesis in mammalian cells. More specifically, mutant strains will be selected which are deficient in various DNA repair pathways. These strains will be studied with regard to (1) the nature of the defect in repair, and (2) the mutability and transformability of the defective cells by various agents as compared to the wild type parental cells. The results to date include progress in the following areas: (1) determination of optimum conditions for growth and maintenance of cells and for quantitative measurement of various cellular parameters; (2) investigation of the effect of holding mutagenized cells for various periods in a density inhibited state on survival and on mutation and transformation frequencies; (3) examination of the repair capabilities of BHK cells, as compared to repair-proficient and repair-deficient human cells and excision-deficient mouse cells, as measured by the reactivation of Herpes simplex virus (HSV) treated with radiation and ethylmethane sulfonate (EMS); (4) initiation of host cell reactivation viral sucide enrichment and screening of survivors of the enrichment for sensitivity to ionizing radiation; and (5) investigation of the toxicity, mutagenicity, and carcinogenicity of various metabolites of 4-nitroquinoline-1-oxide (4-NQO). (ERB)

  1. Managing turbine-generator outages by computer

    SciTech Connect (OSTI)

    Reinhart, E.R. [Reinhart and Associates, Inc., Austin, TX (United States)

    1997-09-01

    This article describes software being developed to address the need for computerized planning and documentation programs that can help manage outages. Downsized power-utility companies and the growing demand for independent, competitive engineering and maintenance services have created a need for a computer-assisted planning and technical-direction program for turbine-generator outages. To meet this need, a software tool is now under development that can run on a desktop or laptop personal computer to assist utility personnel and technical directors in outage planning. Total Outage Planning Software (TOPS), which runs on Windows, takes advantage of the mass data storage available with compact-disc technology by archiving the complete outage documentation on CD. Previous outage records can then be indexed, searched, and viewed on a computer with the click of a mouse. Critical-path schedules, parts lists, parts order tracking, work instructions and procedures, custom data sheets, and progress reports can be generated by computer on-site during an outage.

  2. Protein Co-Expression Network Analysis (ProCoNA)

    SciTech Connect (OSTI)

    Gibbs, David L.; Baratt, Arie; Baric, Ralph; Kawaoka, Yoshihiro; Smith, Richard D.; Orwoll, Eric S.; Katze, Michael G.; Mcweeney, Shannon K.

    2013-06-01

    Biological networks are important for elucidating disease etiology due to their ability to model complex high dimensional data and biological systems. Proteomics provides a critical data source for such models, but currently lacks robust de novo methods for network construction, which could bring important insights in systems biology. We have evaluated the construction of network models using methods derived from weighted gene co-expression network analysis (WGCNA). We show that approximately scale-free peptide networks, composed of statistically significant modules, are feasible and biologically meaningful using two mouse lung experiments and one human plasma experiment. Within each network, peptides derived from the same protein are shown to have a statistically higher topological overlap and concordance in abundance, which is potentially important for inferring protein abundance. The module representatives, called eigenpeptides, correlate significantly with biological phenotypes. Furthermore, within modules, we find significant enrichment for biological function and known interactions (gene ontology and protein-protein interactions). Biological networks are important tools in the analysis of complex systems. In this paper we evaluate the application of weighted co-expression network analysis to quantitative proteomics data. Protein co-expression networks allow novel approaches for biological interpretation, quality control, inference of protein abundance, a framework for potentially resolving degenerate peptide-protein mappings, and a biomarker signature discovery.

  3. Mapping N-linked Glycosylation Sites in the Secretome and Whole Cells of Aspergillus niger Using Hydrazide Chemistry and Mass Spectrometry

    SciTech Connect (OSTI)

    Wang, Lu; Aryal, Uma K.; Dai, Ziyu; Mason, Alisa C.; Monroe, Matthew E.; Tian, Zhixin; Zhou, Jianying; Su, Dian; Weitz, Karl K.; Liu, Tao; Camp, David G.; Smith, Richard D.; Baker, Scott E.; Qian, Weijun

    2012-01-01

    Protein glycosylation is known to play an essential role in both cellular functions and the secretory pathways; however, little information is available on the dynamics of glycosylated N-linked glycosites of fungi. Herein we present the first extensive mapping of glycosylated N-linked glycosites in industrial strain Aspergillus niger by applying an optimized solid phase enrichment of glycopeptide protocol using hydrazide modified magnetic beads. The enrichment protocol was initially optimized using mouse plasma and A. niger secretome samples, which was then applied to profile N-linked glycosites from both the secretome and whole cell lysates of A. niger. A total of 847 unique N-linked glycosites and 330 N-linked glycoproteins were confidently identified by LC-MS/MS. Based on gene ontology analysis, the identified N-linked glycoproteins in the whole cell lysate were primarily localized in the plasma membrane, endoplasmic reticulum, golgi apparatus, lysosome, and storage vacuoles. The identified N-linked glycoproteins are involved in a wide range of biological processes including gene regulation and signal transduction, protein folding and assembly, protein modification and carbohydrate metabolism. The extensive coverage of glycosylated N-linked glycosites along with identification of partial N-linked glycosylation in those enzymes involving in different biochemical pathways provide useful information for functional studies of N-linked glycosylation and their biotechnological applications in A. niger.

  4. Characterization of genetic variability of Venezuelan equine encephalitis viruses

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Gardner, Shea N.; McLoughlin, Kevin; Be, Nicholas A.; Allen, Jonathan; Weaver, Scott C.; Forrester, Naomi; Guerbois, Mathilde; Jaing, Crystal

    2016-04-07

    Venezuelan equine encephalitis virus (VEEV) is a mosquito-borne alphavirus that has caused large outbreaks of severe illness in both horses and humans. New approaches are needed to rapidly infer the origin of a newly discovered VEEV strain, estimate its equine amplification and resultant epidemic potential, and predict human virulence phenotype. We performed whole genome single nucleotide polymorphism (SNP) analysis of all available VEE antigenic complex genomes, verified that a SNP-based phylogeny accurately captured the features of a phylogenetic tree based on multiple sequence alignment, and developed a high resolution genome-wide SNP microarray. We used the microarray to analyze a broadmore » panel of VEEV isolates, found excellent concordance between array- and sequence-based SNP calls, genotyped unsequenced isolates, and placed them on a phylogeny with sequenced genomes. The microarray successfully genotyped VEEV directly from tissue samples of an infected mouse, bypassing the need for viral isolation, culture and genomic sequencing. Lastly, we identified genomic variants associated with serotypes and host species, revealing a complex relationship between genotype and phenotype.« less

  5. Manual for implementing residual radioactive material guidelines using RESRAD, Version 5.0

    SciTech Connect (OSTI)

    Yu, C.; Zielen, A.J.; Cheng, J.J.

    1993-09-01

    This manual presents information for implementing US Department of Energy (DOE) guidelines for residual radioactive material. It describes the analysis and models used to derive site-specific guidelines for allowable residual concentrations of radionuclides in soil and the design and use of the RESRAD computer code for calculating doses, risks, and guideline values. It also describes procedures for implementing DOE policy for reducing residual radioactivity to levels that are as low as reasonably achievable. Two new pathways, radon inhalation and soil ingestion, have been added to RESRAD. Twenty-seven new radionuclides have also been added, and the cutoff half-life for associated radionuclides has been reduced to six months. Other major improvements to the RESRAD code include the ability to run sensitivity analyses, the addition of graphical output, user-specified dose factors, updated databases, an improved groundwater transport model, optional input of a groundwater concentration and a solubility constant, special models for tritium and carbon-14, calculation of cancer incidence risk, and the use of a mouse with menus.

  6. Crystallization and preliminary crystallographic studies of human kallikrein 7, a serine protease of the multigene kallikrein family

    SciTech Connect (OSTI)

    Fernndez, Israel S.; Stndker, Ludger; Forssmann, Wolf-Georg; Gimnez-Gallego, Guillermo; Romero, Antonio

    2007-08-01

    The cloning, expression, purification and crystallization of recombinant human kallikrein 7, directly synthesized in the active form in E. coli, is described. Diffraction data were collected to 2.8 resolution from native crystals. Human kallikreins are a group of serine proteases of high sequence homology whose genes are grouped as a single cluster at chromosome 19. Although the physiological roles of kallikreins are generally still unknown, members of the kallikrein family have been clearly implicated in pathological situations such as cancer and psoriasis. Human kallikrein 7 (hK7) has been shown to be involved in pathological keratinization, psoriasis and ovarian cancer. In order to gain insight into the molecular structure of this protein, hK7 was crystallized after recombinant production in its folded and active form using a periplasmic secretion vector in Escherichia coli. The crystals belonged to the rhombohedral space group H32 and diffracted to 2.8 . The phase problem was solved by molecular replacement using the mouse kallikrein-related protein neuropsin. Completion of the model and structure refinement are under way.

  7. Acoustic Droplet Vaporization, Cavitation, and Therapeutic Properties of Copolymer-Stabilized Perfluorocarbon Nanoemulsions

    SciTech Connect (OSTI)

    Nam, Kweon-Ho; Christensen, Douglas A.; Rapoport, Natalya; Kennedy, Anne M.

    2009-04-14

    Acoustic and therapeutic properties of Doxorubicin (DOX) and paclitaxel (PTX)-loaded perfluorocarbon nanoemulsions have been investigated in a mouse model of ovarian cancer. The nanoemulsions were stabilized by two biodegradable amphiphilic block copolymers that differed in the structure of the hydrophobic block. Acoustic droplet vaporization (ADV) and cavitation parameters were measured as a function of ultrasound frequency, pressure, duty cycles, and temperature. The optimal parameters that induced ADV and inertial cavitation of the formed microbubbles were used in vivo in the experiments on the ultrasound-mediated chemotherapy of ovarian cancer. A combination tumor treatment by intravenous injections of drug-loaded perfluoropentane nanoemulsions and tumor-directed 1-MHz ultrasound resulted in a dramatic decrease of ovarian or breast carcinoma tumor volume and sometimes complete tumor resolution. However, tumors often recurred three to six weeks after the treatment indicating that some cancer cells survived the treatment. The recurrent tumors proved more aggressive and resistant to the repeated therapy than initial tumors suggesting selection for the resistant cells during the first treatment.

  8. Cancer Associated Fibroblasts express pro-inflammatory factors in human breast and ovarian tumors

    SciTech Connect (OSTI)

    Erez, Neta; Glanz, Sarah; Raz, Yael; Department of Obstetrics and Gynecology, LIS Maternity Hospital, Tel Aviv Sourasky Medical Center, affiliated with Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv ; Avivi, Camilla; Barshack, Iris; Department of Pathology, Sheba Medical Center, Tel Hashomer, affiliated with Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv

    2013-08-02

    Highlights: CAFs in human breast and ovarian tumors express pro-inflammatory factors. Expression of pro-inflammatory factors correlates with tumor invasiveness. Expression of pro-inflammatory factors is associated with NF-?b activation in CAFs. -- Abstract: Inflammation has been established in recent years as a hallmark of cancer. Cancer Associated Fibroblasts (CAFs) support tumorigenesis by stimulating angiogenesis, cancer cell proliferation and invasion. We previously demonstrated that CAFs also mediate tumor-enhancing inflammation in a mouse model of skin carcinoma. Breast and ovarian carcinomas are amongst the leading causes of cancer-related mortality in women and cancer-related inflammation is linked with both these tumor types. However, the role of CAFs in mediating inflammation in these malignancies remains obscure. Here we show that CAFs in human breast and ovarian tumors express high levels of the pro-inflammatory factors IL-6, COX-2 and CXCL1, previously identified to be part of a CAF pro-inflammatory gene signature. Moreover, we show that both pro-inflammatory signaling by CAFs and leukocyte infiltration of tumors are enhanced in invasive ductal carcinoma as compared with ductal carcinoma in situ. The pro-inflammatory genes expressed by CAFs are known NF-?B targets and we show that NF-?B is up-regulated in breast and ovarian CAFs. Our data imply that CAFs mediate tumor-promoting inflammation in human breast and ovarian tumors and thus may be an attractive target for stromal-directed therapeutics.

  9. Proliferation of Estrogen Receptor alpha Positive Mammary Epithelial Cells is Restrained by TGFbeta1 in Adult Mice

    SciTech Connect (OSTI)

    Ewan, Kenneth B.R.; Oketch-Rabah, Hellen A.; Ravani, Shraddha A.; Shyamala, G.; Moses, Harold L.; Barcellos-Hoff, Mary Helen

    2005-03-03

    Transforming growth factor {beta}1 (TGF{beta}1) is a potent inhibitor of mammary epithelial proliferation. In human breast, estrogen receptor {alpha} (ER{alpha}) cells rarely co-localize with markers of proliferation, but their increased frequency correlates with breast cancer risk. To determine whether TGF{beta}1 is necessary for the quiescence of ER{alpha}-positive population, we examined mouse mammary epithelial gland at estrus. Approximately 35% of cells showed TGF{beta}1 activation, which co-localized with nuclear receptor-phosphorylated Smad 2/3, indicating that TGF{beta} signaling is autocrine. Furthermore, nuclear Smad co-localized with nuclear ER{alpha}. To test whether TGF{beta} was functional, we examined genetically engineered mice with different levels of TGF{beta}1. ER{alpha} co-localization with markers of proliferation (i.e. Ki-67 or BrdU) at estrus was significantly increased in the mammary glands of Tgf{beta}1 C57/bl/129SV heterozygote mice. This relationship was maintained following pregnancy, but was absent at puberty. Conversely, mammary epithelial expression of constitutively active TGF{beta}1 via the MMTV promoter suppressed proliferation of ER{alpha} positive cells. Thus, TGF{beta}1 activation functionally restrains ER{alpha} positive cells from proliferating in adult mammary gland. Accordingly, we propose that TGF{beta}1 dysregulation may promote proliferation of ER{alpha} positive cells associated with breast cancer risk in humans.

  10. MCM ring hexamerization is a prerequisite for DNA-binding

    SciTech Connect (OSTI)

    Froelich, Clifford A.; Nourse, Amanda; Enemark, Eric J.

    2015-09-13

    The hexameric Minichromosome Maintenance (MCM) protein complex forms a ring that unwinds DNA at the replication fork in eukaryotes and archaea. Our recent crystal structure of an archaeal MCM N-terminal domain bound to single-stranded DNA (ssDNA) revealed ssDNA associating across tight subunit interfaces but not at the loose interfaces, indicating that DNA-binding is governed not only by the DNA-binding residues of the subunits (MCM ssDNA-binding motif, MSSB) but also by the relative orientation of the subunits. We now extend these findings to show that DNA-binding by the MCM N-terminal domain of the archaeal organism Pyrococcus furiosus occurs specifically in the hexameric oligomeric form. We show that mutants defective for hexamerization are defective in binding ssDNA despite retaining all the residues observed to interact with ssDNA in the crystal structure. One mutation that exhibits severely defective hexamerization and ssDNA-binding is at a conserved phenylalanine that aligns with the mouse Mcm4(Chaos3) mutation associated with chromosomal instability, cancer, and decreased intersubunit association.

  11. Enzyme-Directed Assembly of Nanoparticles in Tumors Monitored by In Vivo Whole Animal and Ex Vivo Super-Resolution Fluorescence Imaging

    SciTech Connect (OSTI)

    Chien, Miao-Ping; Carlini, Andrea S.; Hu, Dehong; Barback, Christopher V.; Rush, Anthony M.; Hall, David J.; Orr, Galya; Gianneschi, Nathan C.

    2013-12-18

    Matrix metalloproteinase enzymes, overexpressed in HT-1080 human fibrocarcinoma tumors, were used to guide the accumulation and retention of an enzyme-responsive nanoparticle in a xenograft mouse model. The nanoparticles were prepared as micelles from amphiphilic block copolymers bearing a simple hydrophobic block, and a hydrophilic peptide brush. The polymers were end-labeled with Alexa Fluor 647 dyes leading to the formation of labeled micelles upon dialysis of the polymers from DMSO to aqueous buffer. This dye-labeling strategy allowed the presence of the retained material to be visualized via whole animal imaging in vivo, and in ex vivo organ analysis following intratumoral injection into HT-1080 xenograft tumors. We propose that the material is retained by virtue of an enzyme-induced accumulation process whereby particles change morphology from 20 nm spherical micelles to micron-scale aggregates, kinetically trapping them within the tumor. This hypothesis is tested here via an unprecedented super resolution fluorescence analysis of ex vivo tissue slices confirming a particle size increase occurs concomitantly with extended retention of responsive particles compared to unresponsive controls.

  12. Radiation leukaemogenesis at low doses DE-FG02-05 ER 63947 Final Technical Report 15 May 2005 ???????????????¢???????????????????????????????? 14 May 2010

    SciTech Connect (OSTI)

    Simon Bouffler

    2010-07-28

    This report provides a complete summary of the work undertaken and results obtained under US Department of Energy grant DF-FG02-05 ER 63947, Radiation leukaemogenesis at low doses. There is ample epidemiological evidence indicating that ionizing radiation is carcinogenic in the higher dose range. This evidence, however, weakens and carries increasing uncertainties at doses below 100-200 mSv. At these low dose levels the form of the dose-response curve for radiation-induced cancer cannot be determined reliably or directly from studies of human populations. Therefore animal, cellular and other experimental systems must be employed to provide supporting evidence on which to base judgements of risk at low doses. Currently in radiological protection a linear non-threshold (LNT) extrapolation of risk estimates derived from human epidemiological studies is used to estimate risks in the dose range of interest for protection purposes. Myeloid leukaemias feature prominently among the cancers associated with human exposures to ionising radiation (eg UNSCEAR 2006; IARC 2000). Good animal models of radiation-induced acute myeloid leukaemia (AML) are available including strains such as CBA, RFM and SJL (eg Major and Mole 1978; Ullrich et al 1976; Resnitzky et al 1985). Early mechanistic studies using cytogenetic methods in these mouse models established that the majority of radiation-induced AMLs carried substantial interstitial deletions in one copy of chromosome (chr) 2 (eg Hayata et al 1983; Trakhtenbrot et al 1988; Breckon et al 1991; Rithidech et al 1993; Bouffler et al 1996). Chr2 aberrations are known to occur in bone marrow cells as early as 24 hours after in vivo irradiation (Bouffler et al 1997). Subsequent molecular mapping studies defined a distinct region of chr2 that is commonly lost in AMLs (Clark et al 1996; Silver et al 1999). Further, more detailed, analysis identified point mutations at a specific region of the Sfpi1/PU.1 haemopoietic transcription factor gene which lies in the commonly deleted region of chr2 (Cook et al 2004; Suraweera et al 2005). These lines of evidence strongly implicate the Sfpi1/PU.1 gene as a tumour suppressor gene, dysregulation of which leads to myeloid leukaemia. The main focus of this project was to utilize the CBA mouse model of radiation leukaemogenesis to explore mechanisms of low dose and low dose-rate leukaemogenesis. A series of mechanistic investigations were undertaken, the central aim of which was to identify the events that convert normal cells into myeloid leukaemia cells and explore the dose-response relationships for these. Much of the work centred on the Sfpi1/PU.1 gene and its role in leukaemogenesis. Specific studies considered the dose-response and time-course relationships for loss of the gene, the functional consequences of Sfpi1/PU.1 loss and mutation on transcriptional programmes and developing an in vivo reporter gene system for radiation-induced alterations to PU.1 expression. Additional work sought further genetic changes associated with radiation-induced AMLs and a better characterization of the cell of origin or 'target cell' for radiation-induced AML. All the information gathered is of potential use in developing biologically realistic mathematical models for low dose cancer risk projection.

  13. Trypanosoma Cruzi Cyp51 Inhibitor Derived from a Mycobacterium Tuberculosis Screen Hit

    SciTech Connect (OSTI)

    Chen, Chiung-Kuang; Doyle, Patricia S.; Yermalitskaya, Liudmila V.; Mackey, Zachary B.; Ang, Kenny K.H.; McKerrow, James H.; Podust, Larissa M.

    2009-02-18

    The two front-line drugs for chronic Trypanosoma cruzi infections are limited by adverse side-effects and declining efficacy. One potential new target for Chagas disease chemotherapy is sterol 14{alpha}-demethylase (CYP51), a cytochrome P450 enzyme involved in biosynthesis of membrane sterols. In a screening effort targeting Mycobacterium tuberculosis CYP51 (CYP51{sub Mt}), we previously identified the N-[4-pyridyl]-formamide moiety as a building block capable of delivering a variety of chemotypes into the CYP51 active site. In that work, the binding modes of several second generation compounds carrying this scaffold were determined by high-resolution co-crystal structures with CYP51{sub Mt}. Subsequent assays against the CYP51 orthologue in T. cruzi, CYP51{sub Tc}, demonstrated that two of the compounds tested in the earlier effort bound tightly to this enzyme. Both were tested in vitro for inhibitory effects against T. cruzi and the related protozoan parasite Trypanosoma brucei, the causative agent of African sleeping sickness. One of the compounds had potent, selective anti-T. cruzi activity in infected mouse macrophages. Cure of treated host cells was confirmed by prolonged incubation in the absence of the inhibiting compound. Discrimination between T. cruzi and T. brucei CYP51 by the inhibitor was largely based on the variability (phenylalanine versus isoleucine) of a single residue at a critical position in the active site. CYP51{sub Mt}-based crystal structure analysis revealed that the functional groups of the two tightly bound compounds are likely to occupy different spaces in the CYP51 active site, suggesting the possibility of combining the beneficial features of both inhibitors in a third generation of compounds to achieve more potent and selective inhibition of CYP51{sub Tc}. Enzyme sterol 14{alpha}-demethylase (CYP51) is a well-established target for anti-fungal therapy and is a prospective target for Chagas disease therapy. We previously identified a chemical scaffold capable of delivering a variety of chemical structures into the CYP51 active site. In this work the binding modes of several second generation compounds carrying this scaffold were determined in high-resolution co-crystal structures with CYP51 of Mycobacterium tuberculosis. Subsequent assays against CYP51 in Trypanosoma cruzi, the agent of Chagas disease, demonstrated that two of the compounds bound tightly to the enzyme. Both were tested for inhibitory effects against T. cruzi and the related protozoan parasite Trypanosoma brucei. One of the compounds had potent, selective anti-T. cruzi activity in infected mouse macrophages. This compound is currently being evaluated in animal models of Chagas disease. Discrimination between T. cruzi and T. brucei CYP51 by the inhibitor was largely based on the variability of a single amino acid residue at a critical position in the active site. Our work is aimed at rational design of potent and highly selective CYP51 inhibitors with potential to become therapeutic drugs. Drug selectivity to prevent host-pathogen cross-reactivity is pharmacologically important, because CYP51 is present in human host.

  14. Inhibition of cell proliferation by a selective inhibitor of the Ca{sup 2+}-activated Cl{sup -} channel, Ano1

    SciTech Connect (OSTI)

    Mazzone, Amelia; Eisenman, Seth T.; Strege, Peter R.; Yao, Zhen; Ordog, Tamas; Gibbons, Simon J.; Farrugia, Gianrico

    2012-10-19

    Highlights: Black-Right-Pointing-Pointer T16A{sub inh}-A01 blocked Ano1 currents in HEK cells expressing Ano1. Black-Right-Pointing-Pointer T16A{sub inh}-A01 reduced proliferation in ICC primary cultures and CFPAC-1 cell line. Black-Right-Pointing-Pointer T16A{sub inh}-A01 reduced proliferation of ICC in intact smooth muscle strips. -- Abstract: Background: Ion channels play important roles in regulation of cellular proliferation. Ano1 (TMEM16A) is a Ca{sup 2+}-activated Cl{sup -} channel expressed in several tumors and cell types. In the muscle layers of the gastrointestinal tract Ano1 is selectively expressed in interstitial cells of Cajal (ICC) and appears to be required for normal gastrointestinal slow wave electrical activity. However, Ano1 is expressed in all classes of ICC, including those that do not generate slow waves suggesting that Ano1 may have other functions. Indeed, a role for Ano1 in regulating proliferation of tumors and ICC has been recently suggested. Recently, a high-throughput screen identified a small molecule, T16A{sub inh}-A01 as a specific inhibitor of Ano1. Aim: To investigate the effect of the T16A{sub inh}-A01 inhibitor on proliferation in ICC and in the Ano1-expressing human pancreatic cancer cell line CFPAC-1. Methods: Inhibition of Ano1 was demonstrated by whole cell voltage clamp recordings of currents in cells transfected with full-length human Ano1. The effect of T16A{sub inh}-A01 on ICC proliferation was examined in situ in organotypic cultures of intact mouse small intestinal smooth muscle strips and in primary cell cultures prepared from these tissues. ICC were identified by Kit immunoreactivity. Proliferating ICC and CFPAC-1 cells were identified by immunoreactivity for the nuclear antigen Ki67 or EdU incorporation, respectively. Results: T16A{sub inh}-A01 inhibited Ca{sup 2+}-activated Cl{sup -} currents by 60% at 10 {mu}M in a voltage-independent fashion. Proliferation of ICC was significantly reduced in primary cultures from BALB/c mice following treatment with T16A{sub inh}-A01. Proliferation of the CFPAC-1 human cell-line was also reduced by T16A{sub inh}-A01. In organotypic cultures of smooth muscle strips from mouse jejunum, the proliferation of ICC was reduced but the total number of proliferating cells/confocal stack was not affected, suggesting that the inhibitory effect was specific for ICC. Conclusions: The selective Ano1 inhibitor T16A{sub inh}-A01 inhibited Ca{sup 2+}-activated Cl{sup -} currents, reduced the number of proliferating ICC in culture and inhibited proliferation in the pancreatic cancer cell line CFPAC-1. These data support the notion that chloride channels in general and Ano1 in particular are involved in the regulation of proliferation.

  15. Seed-mediated synthesis, properties and application of {gamma}-Fe{sub 2}O{sub 3}-CdSe magnetic quantum dots

    SciTech Connect (OSTI)

    Lin, Alex W.H.; Ang, Chung Yen; Patra, Pranab K.; Han Yu; Gu Hongwei; Le Breton, Jean-Marie; Juraszek, Jean; Chiron, Hubert; Papaefthymiou, Georgia C.; Tamil Selvan, Subramanian; Ying, Jackie Y.

    2011-08-15

    Seed-mediated growth of fluorescent CdSe quantum dots (QDs) around {gamma}-Fe{sub 2}O{sub 3} magnetic cores was performed at high temperature (300 deg. C) in the presence of organic surfactants. Bi-functional magnetic quantum dots (MQDs) with tunable emission properties were successfully prepared. The as-synthesized MQDs were characterized by high-resolution transmission electron microscopy (HRTEM) and dynamic light scattering (DLS), which confirmed the assembly of heterodimers. When a longer growth period was employed, a homogeneous dispersion of QDs around a magnetic nanoparticle was obtained. The magnetic properties of these nanocomposites were examined. The MQDs were superparamagnetic with a saturation magnetization of 0.40 emu/g and a coercivity of 138 Oe at 5 K. To demonstrate their potential application in bio-labeling, these MQDs were coated with a thin silica shell, and functionalized with a polyethylene glycol (PEG) derivative. The functionalized MQDs were effectively used for the labeling of live cell membranes of 4T1 mouse breast cancer cells and HepG2 human liver cancer cells. - Graphical abstract: (a) HRTEM image of oleic acid capped MPs. The size of MPs ranges from 8 to 10 nm. (b) XRD pattern of {gamma}-Fe{sub 2}O{sub 3} MPs. Highlights: > The fabrication of MQDs through a seed-mediated approach has been demonstrated. > The formation and assembly of these bi-functional nanocomposites have been elucidated. > The MQDs exhibit superparamagnetism and tunable emissions characteristic of the components. > MQDs with thin silica coating were successfully employed in the labeling of cancer cell membranes.

  16. Non-lethal heat treatment of cells results in reduction of tumor initiation and metastatic potential

    SciTech Connect (OSTI)

    Kim, Yoo-Shin; Lee, Tae Hoon; O'Neill, Brian E.

    2015-08-14

    Non-lethal hyperthermia is used clinically as adjuvant treatment to radiation, with mixed results. Denaturation of protein during hyperthermia treatment is expected to synergize with radiation damage to cause cell cycle arrest and apoptosis. Alternatively, hyperthermia is known to cause tissue level changes in blood flow, increasing the oxygenation and radiosensitivity of often hypoxic tumors. In this study, we elucidate a third possibility, that hyperthermia alters cellular adhesion and mechanotransduction, with particular impact on the cancer stem cell population. We demonstrate that cell heating results in a robust but temporary loss of cancer cell aggressiveness and metastatic potential in mouse models. In vitro, this heating results in a temporary loss in cell mobility, adhesion, and proliferation. Our hypothesis is that the loss of cellular adhesion results in suppression of cancer stem cells and loss of tumor virulence and metastatic potential. Our study suggests that the metastatic potential of cancer is particularly reduced by the effects of heat on cellular adhesion and mechanotransduction. If true, this could help explain both the successes and failures of clinical hyperthermia, and suggest ways to target treatments to those who would most benefit. - Highlights: • Non-lethal hyperthermia treatment of cancer cells is shown to cause a reduction in rates of tumor initiation and metastasis. • Dynamic imaging of cells during heat treatment shows temporary changes in cell shape, cell migration, and cell proliferation. • Loss of adhesion may lead to the observed effect, which may disproportionately impact the tumor initiating cell fraction. • Loss or suppression of the tumor initiating cell fraction results in the observed loss of metastatic potential in vivo. • This result may lead to new approaches to synergizing hyperthermia with surgery, radiation, and chemotherapy.

  17. YAP/TAZ enhance mammalian embryonic neural stem cell characteristics in a Tead-dependent manner

    SciTech Connect (OSTI)

    Han, Dasol; Byun, Sung-Hyun; Park, Soojeong; Kim, Juwan; Kim, Inhee; Ha, Soobong; Kwon, Mookwang; Yoon, Keejung

    2015-02-27

    Mammalian brain development is regulated by multiple signaling pathways controlling cell proliferation, migration and differentiation. Here we show that YAP/TAZ enhance embryonic neural stem cell characteristics in a cell autonomous fashion using diverse experimental approaches. Introduction of retroviral vectors expressing YAP or TAZ into the mouse embryonic brain induced cell localization in the ventricular zone (VZ), which is the embryonic neural stem cell niche. This change in cell distribution in the cortical layer is due to the increased stemness of infected cells; YAP-expressing cells were colabeled with Sox2, a neural stem cell marker, and YAP/TAZ increased the frequency and size of neurospheres, indicating enhanced self-renewal- and proliferative ability of neural stem cells. These effects appear to be TEA domain family transcription factor (Tead)–dependent; a Tead binding-defective YAP mutant lost the ability to promote neural stem cell characteristics. Consistently, in utero gene transfer of a constitutively active form of Tead2 (Tead2-VP16) recapitulated all the features of YAP/TAZ overexpression, and dominant negative Tead2-EnR resulted in marked cell exit from the VZ toward outer cortical layers. Taken together, these results indicate that the Tead-dependent YAP/TAZ signaling pathway plays important roles in neural stem cell maintenance by enhancing stemness of neural stem cells during mammalian brain development. - Highlights: • Roles of YAP and Tead in vivo during mammalian brain development are clarified. • Expression of YAP promotes embryonic neural stem cell characteristics in vivo in a cell autonomous fashion. • Enhancement of neural stem cell characteristics by YAP depends on Tead. • Transcriptionally active form of Tead alone can recapitulate the effects of YAP. • Transcriptionally repressive form of Tead severely reduces stem cell characteristics.

  18. Low-frequency quantitative ultrasound imaging of cell death in vivo

    SciTech Connect (OSTI)

    Sadeghi-Naini, Ali; Falou, Omar; Czarnota, Gregory J.; Department of Radiation Oncology, Odette Cancer Centre, Sunnybrook Health Sciences Centre, Toronto, Ontario M4N 3M5; Department of Medical Biophysics, Faculty of Medicine, University of Toronto, Toronto, Ontario M4N 3M5; Department of Radiation Oncology, Faculty of Medicine, University of Toronto, Toronto, Ontario M4N 3M5 ; Papanicolau, Naum; Tadayyon, Hadi; Lee, Justin; Zubovits, Judit; Sadeghian, Alireza; Karshafian, Raffi; Al-Mahrouki, Azza; Giles, Anoja; Kolios, Michael C.

    2013-08-15

    Purpose: Currently, no clinical imaging modality is used routinely to assess tumor response to cancer therapies within hours to days of the delivery of treatment. Here, the authors demonstrate the efficacy of ultrasound at a clinically relevant frequency to quantitatively detect changes in tumors in response to cancer therapies using preclinical mouse models.Methods: Conventional low-frequency and corresponding high-frequency ultrasound (ranging from 4 to 28 MHz) were used along with quantitative spectroscopic and signal envelope statistical analyses on data obtained from xenograft tumors treated with chemotherapy, x-ray radiation, as well as a novel vascular targeting microbubble therapy.Results: Ultrasound-based spectroscopic biomarkers indicated significant changes in cell-death associated parameters in responsive tumors. Specifically changes in the midband fit, spectral slope, and 0-MHz intercept biomarkers were investigated for different types of treatment and demonstrated cell-death related changes. The midband fit and 0-MHz intercept biomarker derived from low-frequency data demonstrated increases ranging approximately from 0 to 6 dBr and 0 to 8 dBr, respectively, depending on treatments administrated. These data paralleled results observed for high-frequency ultrasound data. Statistical analysis of ultrasound signal envelope was performed as an alternative method to obtain histogram-based biomarkers and provided confirmatory results. Histological analysis of tumor specimens indicated up to 61% cell death present in the tumors depending on treatments administered, consistent with quantitative ultrasound findings indicating cell death. Ultrasound-based spectroscopic biomarkers demonstrated a good correlation with histological morphological findings indicative of cell death (r{sup 2}= 0.71, 0.82; p < 0.001).Conclusions: In summary, the results provide preclinical evidence, for the first time, that quantitative ultrasound used at a clinically relevant frequency, in addition to high-frequency ultrasound, can detect tissue changes associated with cell death in vivo in response to cancer treatments.

  19. Imaging Biomarker Dynamics in an Intracranial Murine Glioma Study of Radiation and Antiangiogenic Therapy

    SciTech Connect (OSTI)

    Chung, Caroline; Jalali, Shahrzad; Foltz, Warren; Burrell, Kelly; Wildgoose, Petra; Lindsay, Patricia; Graves, Christian; Camphausen, Kevin; Milosevic, Michael; Jaffray, David; Zadeh, Gelareh; Mnard, Cynthia

    2013-03-01

    Purpose: There is a growing need for noninvasive biomarkers to guide individualized spatiotemporal delivery of radiation therapy (RT) and antiangiogenic (AA) therapy for brain tumors. This study explored early biomarkers of response to RT and the AA agent sunitinib (SU), in a murine intracranial glioma model, using serial magnetic resonance imaging (MRI). Methods and Materials: Mice with MRI-visible tumors were stratified by tumor size into 4 therapy arms: control, RT, SU, and SU plus RT (SURT). Single-fraction conformal RT was delivered using MRI and on-line cone beam computed tomography (CT) guidance. Serial MR images (T2-weighted, diffusion, dynamic contrast-enhanced and gadolinium-enhanced T1-weighted scans) were acquired biweekly to evaluate tumor volume, apparent diffusion coefficient (ADC), and tumor perfusion and permeability responses (K{sub trans}, K{sub ep}). Results: Mice in all treatment arms survived longer than those in control, with a median survival of 35 days for SURT (P<.0001) and 30 days for RT (P=.009) and SU (P=.01) mice vs 26 days for control mice. At Day 3, ADC rise was greater with RT than without (P=.002). Sunitinib treatment reduced tumor perfusion/permeability values with mean K{sub trans} reduction of 27.6% for SU (P=.04) and 26.3% for SURT (P=.04) mice and mean K{sub ep} reduction of 38.1% for SU (P=.01) and 27.3% for SURT (P=.02) mice. The magnitude of individual mouse ADC responses at Days 3 and 7 correlated with subsequent tumor growth rate R values of ?0.878 (P=.002) and ?0.80 (P=.01), respectively. Conclusions: Early quantitative changes in diffusion and perfusion MRI measures reflect treatment responses soon after starting therapy and thereby raise the potential for these imaging biomarkers to guide adaptive and potentially individualized therapy approaches in the future.

  20. Structural basis for recognition of hemi-methylated DNA by the SRA domain of human UHRF1.

    SciTech Connect (OSTI)

    Avvakumov, George V.; Walker, John R.; Xue, Sheng; Li, Yanjun; Duan, Shili; Bronner, Christian; Arrowsmith, Cheryl H.; Dhe-Paganon, Sirano

    2008-11-17

    Epigenetic inheritance in mammals is characterized by high-fidelity replication of CpG methylation patterns during development. UHRF1 (also known as ICBP90 in humans and Np95 in mouse) is an E3 ligase important for the maintenance of global and local DNA methylation in vivo. The preferential affinity of UHRF1 for hemi-methylated DNA over symmetrically methylated DNA by means of its SET and RING-associated (SRA) domain and its association with the maintenance DNA methyltransferase 1 (DNMT1) suggests a role in replication of the epigenetic code. Here we report the 1.7 {angstrom} crystal structure of the apo SRA domain of human UHRF1 and a 2.2 {angstrom} structure of its complex with hemi-methylated DNA, revealing a previously unknown reading mechanism for methylated CpG sites (mCpG). The SRA-DNA complex has several notable structural features including a binding pocket that accommodates the 5-methylcytosine that is flipped out of the duplex DNA. Two specialized loops reach through the resulting gap in the DNA from both the major and the minor grooves to read the other three bases of the CpG duplex. The major groove loop confers both specificity for the CpG dinucleotide and discrimination against methylation of deoxycytidine of the complementary strand. The structure, along with mutagenesis data, suggests how UHRF1 acts as a key factor for DNMT1 maintenance methylation through recognition of a fundamental unit of epigenetic inheritance, mCpG.