National Library of Energy BETA

Sample records for human skin cells

  1. p53 modulates the AMPK inhibitor compound C induced apoptosis in human skin cancer cells

    SciTech Connect (OSTI)

    Huang, Shi-Wei [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Wu, Chun-Ying [Division of Gastroenterology and Hepatology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Wang, Yen-Ting [Department of Medical Research and Education, Cheng Hsin General Hospital, Taipei, Taiwan (China); Kao, Jun-Kai [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Department of Pediatrics, Children's Hospital, Changhua Christian Hospital, Changhua, Taiwan (China); Lin, Chi-Chen; Chang, Chia-Che; Mu, Szu-Wei; Chen, Yu-Yu [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Chiu, Husan-Wen [Institute of Biotechnology, National Cheng-Kung University, Tainan, Taiwan (China); Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan (China); Chang, Chuan-Hsun [Department of Surgical Oncology, Cheng Hsin General Hospital, Taipei, Taiwan (China); Department of Nutrition Therapy, Cheng Hsin General Hospital, Taipei, Taiwan (China); School of Nutrition and Health Sciences, Taipei Medical University, Taipei, Taiwan (China); Liang, Shu-Mei [Institute of Biotechnology, National Cheng-Kung University, Tainan, Taiwan (China); Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan (China); Chen, Yi-Ju [Department of Dermatology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Huang, Jau-Ling [Department of Bioscience Technology, Chang Jung Christian University, Tainan, Taiwan (China); Shieh, Jeng-Jer, E-mail: shiehjj@vghtc.gov.tw [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Department of Education and Research, Taichung Veterans General Hospital, Taichung, Taiwan (China)

    2013-02-15

    Compound C, a well-known inhibitor of the intracellular energy sensor AMP-activated protein kinase (AMPK), has been reported to cause apoptotic cell death in myeloma, breast cancer cells and glioma cells. In this study, we have demonstrated that compound C not only induced autophagy in all tested skin cancer cell lines but also caused more apoptosis in p53 wildtype skin cancer cells than in p53-mutant skin cancer cells. Compound C can induce upregulation, phosphorylation and nuclear translocalization of the p53 protein and upregulate expression of p53 target genes in wildtype p53-expressing skin basal cell carcinoma (BCC) cells. The changes of p53 status were dependent on DNA damage which was caused by compound C induced reactive oxygen species (ROS) generation and associated with activated ataxia-telangiectasia mutated (ATM) protein. Using the wildtype p53-expressing BCC cells versus stable p53-knockdown BCC sublines, we present evidence that p53-knockdown cancer cells were much less sensitive to compound C treatment with significant G2/M cell cycle arrest and attenuated the compound C-induced apoptosis but not autophagy. The compound C induced G2/M arrest in p53-knockdown BCC cells was associated with the sustained inactive Tyr15 phosphor-Cdc2 expression. Overall, our results established that compound C-induced apoptosis in skin cancer cells was dependent on the cell's p53 status. - Highlights: ? Compound C caused more apoptosis in p53 wildtype than p53-mutant skin cancer cells. ? Compound C can upregulate p53 expression and induce p53 activation. ? Compound C induced p53 effects were dependent on ROS induced DNA damage pathway. ? p53-knockdown attenuated compound C-induced apoptosis but not autophagy. ? Compound C-induced apoptosis in skin cancer cells was dependent on p53 status.

  2. Green tea polyphenol, (?)-epigallocatechin-3-gallate, induces toxicity in human skin cancer cells by targeting ?-catenin signaling

    SciTech Connect (OSTI)

    Singh, Tripti; Katiyar, Santosh K.

    2013-12-01

    The green tea polyphenol, (?)-epigallocatechin-3-gallate (EGCG), has been shown to have anti-carcinogenic effects in several skin tumor models, and efforts are continued to investigate the molecular targets responsible for its cytotoxic effects to cancer cells. Our recent observation that ?-catenin is upregulated in skin tumors suggested the possibility that the anti-skin carcinogenic effects of EGCG are mediated, at least in part, through its effects on ?-catenin signaling. We have found that treatment of the A431 and SCC13 human skin cancer cell lines with EGCG resulted in reduced cell viability and increased cell death and that these cytotoxic effects were associated with inactivation of ?-catenin signaling. Evidence of EGCG-induced inactivation of ?-catenin included: (i) reduced accumulation of nuclear ?-catenin; (ii) enhanced levels of casein kinase1?, reduced phosphorylation of glycogen synthase kinase-3?, and increased phosphorylation of ?-catenin on critical serine{sup 45,33/37} residues; and (iii) reduced levels of matrix metalloproteinase (MMP)-2 and MMP-9, which are down-stream targets of ?-catenin. Treatment of cells with prostaglandin E2 (PGE{sub 2}) enhanced the accumulation of ?-catenin and enhanced ?-catenin signaling. Treatment with either EGCG or an EP2 antagonist (AH6809) reduced the PGE{sub 2}-enhanced levels of cAMP, an upstream regulator of ?-catenin. Inactivation of ?-catenin by EGCG resulted in suppression of cell survival signaling proteins. siRNA knockdown of ?-catenin in A431 and SCC13 cells reduced cell viability. Collectively, these data suggest that induction of cytotoxicity in skin cancer cells by EGCG is mediated by targeting of ?-catenin signaling and that the ?-catenin signaling is upregulated by inflammatory mediators. - Highlights: • EGCG inhibits cancer cell viability through inactivation of ?-catenin signaling. • Inactivation of ?-catenin involves the downregulation of inflammatory mediators. • EGCG inactivates ?-catenin in skin cancer cells by inhibition of cAMP and PGE{sub 2}. • siRNA knockdown of ?-catenin or COX-2 reduces the viability of cancer cells.

  3. Chemoprevention of human skin cancers.

    E-Print Network [OSTI]

    Loescher, L J; Meyskens, F L Jr

    1991-01-01

    chemopreventive activity of resveratrol, a natural productJM, Dannenberg AJ. Resveratrol inhibits cyclooxygenase-2C, Ma WY, Goranson A, Dong Z. Resveratrol sup- presses cell

  4. Ultraviolet-light-induced transformation of human primary cells

    SciTech Connect (OSTI)

    Sutherland, B.M.

    1981-01-01

    The development of model systems for probing the ultraviolet radiation induced oncogenic transformation of human skin cells is described. (ACR)

  5. In vitro dermal absorption of pyrethroid pesticides in human and rat skin

    SciTech Connect (OSTI)

    Hughes, Michael F.; Edwards, Brenda C.

    2010-07-15

    Dermal exposure to pyrethroid pesticides can occur during manufacture and application. This study examined the in vitro dermal absorption of pyrethroids using rat and human skin. Dermatomed skin from adult male Long Evans rats or human cadavers was mounted in flow-through diffusion cells, and radiolabeled bifenthrin, deltamethrin or cis-permethrin was applied in acetone to the skin. Fractions of receptor fluid were collected every 4 h. At 24 h, the skins were washed with soap and water to remove unabsorbed chemical. The skin was then solubilized. Two additional experiments were performed after washing the skin; the first was tape-stripping the skin and the second was the collection of receptor fluid for an additional 24 h. Receptor fluid, skin washes, tape strips and skin were analyzed for radioactivity. For rat skin, the wash removed 53-71% of the dose and 26-43% remained in the skin. The cumulative percentage of the dose at 24 h in the receptor fluid ranged from 1 to 5%. For human skin, the wash removed 71-83% of the dose and 14-25% remained in the skin. The cumulative percentage of the dose at 24 h in the receptor fluid was 1-2%. Tape-stripping removed 50-56% and 79-95% of the dose in rat and human skin, respectively, after the wash. From 24-48 h, 1-3% and about 1% of the dose diffused into the receptor fluid of rat and human skin, respectively. The pyrethroids bifenthrin, deltamethrin and cis-permethrin penetrated rat and human skin following dermal application in vitro. However, a skin wash removed 50% or more of the dose from rat and human skin. Rat skin was more permeable to the pyrethroids than human skin. Of the dose in skin, 50% or more was removed by tape-stripping, suggesting that permeation of pyrethroids into viable tissue could be impeded. The percentage of the dose absorbed into the receptor fluid was considerably less than the dose in rat and human skin. Therefore, consideration of the skin type used and fractions analyzed are important when using in vitro dermal absorption data for risk assessment.

  6. Skewed distribution of natural killer cells in psoriasis skin lesions

    E-Print Network [OSTI]

    2013-01-01

    of natural killer cells in psoriasis skin lesions Mariana D.and unaf- fected skin of psoriasis patients and normal skinComparisons are made between psoriasis groups (lesional and

  7. Screen-Space Perceptual Rendering of Human Skin JORGE JIMENEZ

    E-Print Network [OSTI]

    Gutierrez, Diego

    SUNDSTEDT Trinity College Dublin and DIEGO GUTIERREZ Universidad de Zaragoza We propose a novel skin shader., Sundstedt, V., and Gutierrez, D. 2009. Screen-Space perceptual rendering of human skin. ACM Trans. Appl.Sundstedt@cs.tcd.ie; D. Gutierrez, Departamento de Inform´atica e Ingenier´ia de Sistemas, Universidad de Zaragoza

  8. Master Project in Stem Cell Biology In Vivo Manipulation of Skin Stem Cells

    E-Print Network [OSTI]

    Uppsala Universitet

    Master Project in Stem Cell Biology ­ In Vivo Manipulation of Skin Stem Cells Our lab is interested in understanding how stem cells contribute to tissue homeostasis and disease. Our model system is currently the skin, which harbors several distinct pools of stem cells. In order to identify regulatory networks

  9. Technical Challenges in Using Human Induced Pluripotent Stem Cells to Model Disease

    E-Print Network [OSTI]

    Saha, Krishanu

    Reprogramming of human somatic cells uses readily accessible tissue, such as skin or blood, to generate embryonic-like induced pluripotent stem cells (iPSCs). This procedure has been applied to somatic cells from patients ...

  10. Skin electronics is one of the most promising applications of stretchable electronics. The versatility of skin electronics can only be guaranteed when it has conformal contact with human

    E-Print Network [OSTI]

    Skin electronics is one of the most promising applications of stretchable electronics. The versatility of skin electronics can only be guaranteed when it has conformal contact with human skin. While analysis for the conformability of skin electronics, including modeling, meshing method and step setup etc

  11. Development of a method for assessing non-targeted radiation damage in an artificial 3D human skin model

    E-Print Network [OSTI]

    Brenner, David Jonathan

    at doses as low as 0.1 Gy in the directly irradiated as well as in the bystander cells. Conclusions. #12;critical relevance in low-dose and/or non-Development of a method for assessing non-targeted radiation damage in an artificial 3D human skin

  12. Deep sequencing of small RNAs from human skin reveals major alterations

    E-Print Network [OSTI]

    Zhang, Weixiong

    Deep sequencing of small RNAs from human skin reveals major alterations in the psoriasis mi Psoriasis is a chronic and complex inflammatory skin disease with lesions displaying dramatically altered m for functional characterization of miRNAs in healthy and diseased skin. INTRODUCTION Psoriasis (PS) is a chronic

  13. 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.

  14. The skin's role in human thermoregulation and comfort

    E-Print Network [OSTI]

    Arens, Edward A; Zhang, H.

    2006-01-01

    the convective and radiation heat transfer are about equal (heat transfer at the skin surface (via conduction, convection, and radiation (Heat transfer through and above the skin. outdoors, wind strongly affects convective heat loss or gain, and radiation (

  15. The Effects of Low Dose Irradiation on Inflammatory Response Proteins in a 3D Reconstituted Human Skin Tissue Model

    SciTech Connect (OSTI)

    Varnum, Susan M.; Springer, David L.; Chaffee, Mary E.; Lien, Katie A.; Webb-Robertson, Bobbie-Jo M.; Waters, Katrina M.; Sacksteder, Colette A.

    2012-12-01

    Skin responses to moderate and high doses of ionizing radiation include the induction of DNA repair, apoptosis, and stress response pathways. Additionally, numerous studies indicate that radiation exposure leads to inflammatory responses in skin cells and tissue. However, the inflammatory response of skin tissue to low dose radiation (<10 cGy) is poorly understood. In order to address this, we have utilized a reconstituted human skin tissue model (MatTek EpiDerm FT) and assessed changes in 23 cytokines twenty-four and forty eight hours following treatment of skin with either 3 or 10 cGy low-dose of radiation. Three cytokines, IFN-?, IL-2, MIP-1?, were significantly altered in response to low dose radiation. In contrast, seven cytokines were significantly altered in response to a high radiation dose of 200 cGy (IL-2, IL-10, IL-13, IFN-?, MIP-1?, TNF ?, and VEGF) or the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (G-CSF, GM-CSF, IL-1?, IL-8, MIP-1?, MIP-1?, RANTES). Additionally, radiation induced inflammation appears to have a distinct cytokine response relative to the non-radiation induced stressor, TPA. Overall, these results indicate that there are subtle changes in the inflammatory protein levels following exposure to low dose radiation and this response is a sub-set of what is seen following a high dose in a human skin tissue model.

  16. Skin strain analysis software for the study of human skin deformation

    E-Print Network [OSTI]

    Marecki, Andrew T. (Andrew Thomas)

    2012-01-01

    Skin strain studies have never been conducted in a precise and automated fashion. Previous in vivo strain investigations have been labor intensive and the data resolution was extremely limited such that their results were ...

  17. The effect of low dose ionizing radiation on homeostasis and functional integrity in an organotypic human skin model

    SciTech Connect (OSTI)

    von Neubeck, Claere; Geniza, Matthew; Kauer, Paula M.; Robinson, Joseph E.; Chrisler, William B.; Sowa, Marianne B.

    2015-05-01

    Outside the protection of earth’s atmosphere, astronauts are exposed to low doses of high linear energy transfer (LET) radiation. Future NASA plans for deep space missions or a permanent settlement on the moon are limited by the health risks associated with space radiation exposures. There is a paucity of direct epidemiological data for low dose exposures to space radiation-relevant high LET ions. Health risk models are used to estimate the risk for such exposures, though these models are based on high dose experiments. There is increasing evidence, however, that low and high dose exposures result in different signaling events at the molecular level, and may involve different response mechanisms. Further, despite their low abundance, high LET particles have been identified as the major contributor to health risk during manned space flight. The human skin is exposed in every external radiation scenario, making it an ideal epithelial tissue model in which to study radiation induced effects. Here, we exposed an in vitro three dimensional (3-D) human organotypic skin tissue model to low doses of high LET oxygen (O), silicon (Si) and iron (Fe) ions. We measured proliferation and differentiation profiles in the skin tissue and examined the integrity of the skin’s barrier function. We discuss the role of secondary particles in changing the proportion of cells receiving a radiation dose, emphasizing the possible impact on radiation-induced health issues in astronauts.

  18. Differentiated human stem cells resemble fetal, not adult, ? cells

    E-Print Network [OSTI]

    Hrvatin, Sinisa

    Human pluripotent stem cells (hPSCs) have the potential to generate any human cell type, and one widely recognized goal is to make pancreatic ? cells. To this end, comparisons between differentiated cell types produced in ...

  19. Thermal analysis of ultrathin, compliant sensors for characterization of the human skin

    E-Print Network [OSTI]

    Rogers, John A.

    and identifying optimized designs. This paper presents an analytical model, validated by the finite element method that have the ability to bend, twist, and stretch like human skin, with minimal effect on natural processes onto a thin, low elastic modulus (60 mm thick, 30 kPa) elastomeric sheet. A cross-sectional view

  20. EVALUATION OF SUB-ZERO AND RESIDENCE TIMES AFTER CONTINUOUS VERSUS MULTIPLE INTERMITTENT CRYOGEN SPRAY COOLING EXPOSURE ON HUMAN SKIN

    E-Print Network [OSTI]

    Aguilar, Guillermo

    ; (2) high heat transfer rates as cryogen is deposited onto the skin and evaporates at the sprayed SPRAY COOLING EXPOSURE ON HUMAN SKIN PHANTOM Julio C. Ramirez-San-Juan Beckman Laser Institute Guillermo Aguilar Department of Mechanical Engineering, University of California Riverside ABSTRACT Cryogen

  1. Quantitative Proteomic Profiling of Low Dose Ionizing Radiation Effects in a Human Skin Model

    SciTech Connect (OSTI)

    Hengel, Shawna; Aldrich, Joshua T.; Waters, Katrina M.; Pasa-Tolic, Ljiljana; Stenoien, David L.

    2014-07-29

    To assess molecular responses to low doses of radiation that may be encountered during medical diagnostic procedures, nuclear accidents, or terrorist acts, a quantitative global proteomic approach was used to identify protein alterations in a reconstituted human skin tissue treated with 10 cGy of ionizing radiation. Subcellular fractionation was employed to remove highly abundant structural proteins and provide insight on radiation induced alterations in protein abundance and localization. In addition, peptides were post-fractionated using high resolution 2-dimensional liquid chromatography to increase the dynamic range of detection of protein abundance and translocation changes. Quantitative data was obtained by labeling peptides with 8-plex isobaric iTRAQ tags. A total of 207 proteins were detected with statistically significant alterations in abundance and/or subcellular localization compared to sham irradiated tissues. Bioinformatics analysis of the data indicated that the top canonical pathways affected by low dose radiation are related to cellular metabolism. Among the proteins showing alterations in abundance, localization and proteolytic processing was the skin barrier protein filaggrin which is consistent with our previous observation that ionizing radiation alters profilaggrin processing with potential effects on skin barrier functions. In addition, a large number of proteases and protease regulators were affected by low dose radiation exposure indicating that altered proteolytic activity may be a hallmark of low dose radiation exposure. While several studies have demonstrated altered transcriptional regulation occurs following low dose radiation exposures, the data presented here indicates post-transcriptional regulation of protein abundance, localization, and proteolytic processing play an important role in regulating radiation responses in complex human tissues.

  2. Effects of Wnt3a on proliferation and differentiation of human epidermal stem cells

    SciTech Connect (OSTI)

    Jia Liwei; Zhou Jiaxi; Peng Sha; Li Juxue [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing 100101 (China); Graduate School of the Chinese Academy of Sciences, 19 Yu-quan Road, Beijing 100039 (China); Cao Yujing [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing 100101 (China); Duan Enkui [State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing 100101 (China)], E-mail: duane@ioz.ac.cn

    2008-04-11

    Epidermal stem cells maintain development and homeostasis of mammalian epidermis throughout life. However, the molecular mechanisms involved in the proliferation and differentiation of epidermal stem cells are far from clear. In this study, we investigated the effects of Wnt3a and Wnt/{beta}-catenin signaling on proliferation and differentiation of human fetal epidermal stem cells. We found both Wnt3a and active {beta}-catenin, two key members of the Wnt/{beta}-catenin signaling, were expressed in human fetal epidermis and epidermal stem cells. In addition, Wnt3a protein can promote proliferation and inhibit differentiation of epidermal stem cells in vitro culture. Our results suggest that Wnt/{beta}-catenin signaling plays important roles in human fetal skin development and homeostasis, which also provide new insights on the molecular mechanisms of oncogenesis in human epidermis.

  3. Reflective Terahertz Imaging for early diagnosis of skin burn severity

    E-Print Network [OSTI]

    TEWARI, PRIYAMVADA

    2013-01-01

    of human skin was used for hydration experiments whereby itthe human skin, was used for the hydration experiments and

  4. Generation of Cardiomyocytes from Human Endogenous and Pluripotent Stem-Cell Derived Endothelial Cells

    E-Print Network [OSTI]

    Truong, Raymond

    2013-01-01

    Embryonic Stem Cell Lines Derived from Human Blastocysts.Human Embryonic Stem Cell-Derived Oligodendrocyte ProgenitorPluripotent Stem Cell-Derived Cardiovascular Progenitor

  5. Photoreactivation in bacteria and in skin

    SciTech Connect (OSTI)

    Sutherland, B M

    1980-01-01

    In many procaryotic and eucaryotic cells, photoreactivating enzyme mediates light-dependent repair of uv-induced damage; the enzyme binds to a pyrimidine dimer in DNA, and, on absorption of a photon (300 to 600 nm), specifically monomerizes the dimer, thus repairing the DNA. Photoreactivating enzyme has been found in human tissues and human cells in culture can photoreactivate cellular dimers, and can mediate photoreactivation of Herpes (human fibroblasts) and Epstein-Barr virus (human leukocytes). Measurements of pyrimidine dimer formation and repair in human skin indicate that detectable numbers of dimers are formed at 1 minimal erythemal dose, that the dimiers are rapidly removed in skin kept in the absence of light, and they are more rapidly removed when the skin is exposed to visible light. Whether this apparent photorecovery is true, enzymatic photoreactivation is yet to be determined.

  6. Intrinsic radiation resistance in human chondrosarcoma cells

    SciTech Connect (OSTI)

    Moussavi-Harami, Farid [Departments of Orthopaedics and Rehabilitation, University of Iowa, Iowa City, IA 52242 (United States); Mollano, Anthony [Departments of Orthopaedics and Rehabilitation, University of Iowa, Iowa City, IA 52242 (United States); Martin, James A. [Departments of Orthopaedics and Rehabilitation, University of Iowa, Iowa City, IA 52242 (United States); Ayoob, Andrew [Departments of Orthopaedics and Rehabilitation, University of Iowa, Iowa City, IA 52242 (United States); Domann, Frederick E. [Department of Radiation Oncology, Iowa City, IA 52245 (United States); Gitelis, Steven [Department of Internal Medicine, Section of Medical Oncology, Chicago, IL 60612 (United States); Department of Orthopaedics Rush-Presbyterian St. Luke's Medical Center, Chicago, IL 60612 (United States); Buckwalter, Joseph A. [Departments of Orthopaedics and Rehabilitation, University of Iowa, Iowa City, IA 52242 (United States)]. E-mail: joseph-buckwalter@uiowa.edu

    2006-07-28

    Human chondrosarcomas rarely respond to radiation treatment, limiting the options for eradication of these tumors. The basis of radiation resistance in chondrosarcomas remains obscure. In normal cells radiation induces DNA damage that leads to growth arrest or death. However, cells that lack cell cycle control mechanisms needed for these responses show intrinsic radiation resistance. In previous work, we identified immortalized human chondrosarcoma cell lines that lacked p16{sup ink4a}, one of the major tumor suppressor proteins that regulate the cell cycle. We hypothesized that the absence of p16{sup ink4a} contributes to the intrinsic radiation resistance of chondrosarcomas and that restoring p16{sup ink4a} expression would increase their radiation sensitivity. To test this we determined the effects of ectopic p16{sup ink4a} expression on chondrosarcoma cell resistance to low-dose {gamma}-irradiation (1-5 Gy). p16{sup ink4a} expression significantly increased radiation sensitivity in clonogenic assays. Apoptosis did not increase significantly with radiation and was unaffected by p16{sup ink4a} transduction of chondrosarcoma cells, indicating that mitotic catastrophe, rather than programmed cell death, was the predominant radiation effect. These results support the hypothesis that p16{sup ink4a} plays a role in the radiation resistance of chondrosarcoma cell lines and suggests that restoring p16 expression will improve the radiation sensitivity of human chondrosarcomas.

  7. SHORT REPORT Micropatterning of human embryonic stem cells

    E-Print Network [OSTI]

    Zandstra, Peter W.

    SHORT REPORT Micropatterning of human embryonic stem cells dissects the mesoderm and endoderm in revised form 10 November 2008; accepted 13 November 2008 Abstract Human pluripotent cells such as human embryonic stem cells (hESC) are a great potential source of cells for cell-based therapies; however

  8. Endothelial influences enhance human pluripotent stem cell -derived cardiomyocyte maturation

    E-Print Network [OSTI]

    Wei, Karen A.

    2011-01-01

    Human embryonic stem cell-derived cardiomyocytes survive andet al. , Cardiomyocytes derived from human embryonic stemcardiac function by hESC-derived cardiomyocytes correlates

  9. Thermal Modeling and Experimental Validation of Human Hair and Skin Heated by Broadband Light

    E-Print Network [OSTI]

    Aguilar, Guillermo

    distribution within the hair follicle is highly non-uniform: the minimum temperature occurs at the follicle Sun, PhD,1 Alex Chaney,1 Robert Anderson, PhD,2 and Guillermo Aguilar, PhD 1 * 1 Department:(a)determinetheoveralleffectofPPxonskinhumidi- tyandassociatedskinopticalproperties,and;(b)developaPT numerical model to study the spatial and temporal hair and skin temperature

  10. Changes in Women’s Facial Skin Color Over the Ovulatory Cycle are Not Detectable by the Human Visual System

    E-Print Network [OSTI]

    Burriss, Robert P.; Troscianko, Jolyon; Lovell, P. George; Fulford, Anthony J. C.; Stevens, Martin; Quigley, Rachael; Payne, Jenny; Saxton, Tamsin K.; Rowland, Hannah M.

    2015-01-01

    the mean RGB values for each patch, and converted these to photon 197 catch values equivalent to long, medium, and short wave (LMS) cone responses, and to CIE 198 XYZ responses. We then averaged the left and right patch values, giving one color value per... 199 photograph. Cone-catch models were generated following the methodology of Párraga et al. 200 Running head: FACIAL SKIN COLOR AND THE OVULATORY CYCLE 11 [59]. Human cone-catch quanta (LMS sensitivities from Stockman and Sharpe [71], and CIE 201...

  11. Osmotic water permeability of human red cells

    SciTech Connect (OSTI)

    Terwilliger, T.C.; Solomon, A.K.

    1981-05-01

    The osmotic water permeability of human red cells has been reexamined with a stopped-flow device and a new perturbation technique. Small osmotic gradients are used to minimize the systematic error caused by nonlinearities in the relationship between cell volume and light scattering. Corrections are then made for residual systematic error. Our results show that the hydraulic conductivity, Lp, is essentially independent of the direction of water flow and of osmolality in the range 184-365 mosM. the mean value of Lp obtained obtained was 1.8 +/- 0.1 (SEM) X 10-11 cm3 dyne -1 s-1.

  12. Generation of Human Embryonic Stem Cell-Derived Mesoderm and Cardiac Cells

    E-Print Network [OSTI]

    Zandstra, Peter W.

    ARTICLE Generation of Human Embryonic Stem Cell-Derived Mesoderm and Cardiac Cells Using Size pluripotent stem cell-derived cell types at sufficiently high numbers and in a reproducible manner for the differentiation of pluripotent cells such as human embryonic stem cells (hESC) rely on the generation

  13. High and Low Doses of Ionizing Radiation Induce Different Secretome Profiles in a Human Skin Model

    SciTech Connect (OSTI)

    Zhang, Qibin; Matzke, Melissa M.; Schepmoes, Athena A.; Moore, Ronald J.; Webb-Robertson, Bobbie-Jo M.; Hu, Zeping; Monroe, Matthew E.; Qian, Weijun; Smith, Richard D.; Morgan, William F.

    2014-03-18

    It is postulated that secreted soluble factors are important contributors of bystander effect and adaptive responses observed in low dose ionizing radiation. Using multidimensional liquid chromatography-mass spectrometry based proteomics, we quantified the changes of skin tissue secretome – the proteins secreted from a full thickness, reconstituted 3-dimensional skin tissue model 48 hr after exposure to 3, 10 and 200 cGy of X-rays. Overall, 135 proteins showed statistical significant difference between the sham (0 cGy) and any of the irradiated groups (3, 10 or 200 cGy) on the basis of Dunnett adjusted t-test; among these, 97 proteins showed a trend of downregulation and 9 proteins showed a trend of upregulation with increasing radiation dose. In addition, there were 21 and 8 proteins observed to have irregular trends with the 10 cGy irradiated group either having the highest or the lowest level among all three radiated doses. Moreover, two proteins, carboxypeptidase E and ubiquitin carboxyl-terminal hydrolase isozyme L1 were sensitive to ionizing radiation, but relatively independent of radiation dose. Conversely, proteasome activator complex subunit 2 protein appeared to be sensitive to the dose of radiation, as rapid upregulation of this protein was observed when radiation doses were increased from 3, to 10 or 200 cGy. These results suggest that different mechanisms of action exist at the secretome level for low and high doses of ionizing radiation.

  14. Live cell imaging analysis of the epigenetic regulation of the human endothelial cell migration at single-cell resolution{

    E-Print Network [OSTI]

    Huang, Yanyi

    into individual blank regions, we can measure the migration speed parameter for human primary cells within a few genes. We showed that the migration behaviour of these human primary cells are clearly regulatedLive cell imaging analysis of the epigenetic regulation of the human endothelial cell migration

  15. On the deformation of human skin for mechanical counter pressure space suit development

    E-Print Network [OSTI]

    Obropta, Edward William, Jr

    2015-01-01

    Exploration of planetary bodies requires space suits that do not inhibit astronaut mobility. Gas pressurized suits are typically bulky and stiff to operate or require unnatural human motion. Development of mechanical counter ...

  16. Isolation of stem cells from adult telogen skin Elizabeth Deschene Greco Lab

    E-Print Network [OSTI]

    Greco, Valentina

    in the CO2 chamber · If you are using a transgenic fluorescent mice, verify at the dissection microscope it with a 25 ml plastic pipette for about 1 minute per skin. · Filter the trypsin fraction first with 40 micron(-) serum (chelexed serum) on a shaker. When you have filtered the trypsin fraction you can filter this 10

  17. Gene Expression in Skin and Lymphoblastoid Cells: Refined Statistical Method Reveals

    E-Print Network [OSTI]

    Abecasis, Goncalo

    Rajan P. Nair,2 James T. Elder,2,6 and Gonc¸alo R. Abecasis1,* Psoriasis, an immune to be associated with psoriasis than are randomly selected SNPs. To explore the tissue specificity of these e on psoriasis and other skin traits. Introduction Transcriptional regulation of gene expression is essential

  18. Transgenic rats overexpressing the human MrgX3 gene show cataracts and an abnormal skin phenotype

    SciTech Connect (OSTI)

    Kaisho, Yoshihiko . E-mail: Kaisho_Yoshihiko@takeda.co.jp; Watanabe, Takuya; Nakata, Mitsugu; Yano, Takashi; Yasuhara, Yoshitaka; Shimakawa, Kozo; Mori, Ikuo; Sakura, Yasufumi; Terao, Yasuko; Matsui, Hideki; Taketomi, Shigehisa

    2005-05-13

    The human MrgX3 gene, belonging to the mrgs/SNSRs (mass related genes/sensory neuron specific receptors) family, was overexpressed in transgenic rats using the actin promoter. Two animal lines showed cataracts with liquification/degeneration and swelling of the lens fiber cells. The transient epidermal desquamation was observed in line with higher gene expression. Histopathology of the transgenic rats showed acanthosis and focal parakeratosis. In the epidermis, there was an increase in cellular keratin 14, keratin 10, and loricrin, as well as PGP 9.5 in innervating nerve fibers. These phenotypes accompanied an increase in the number of proliferating cells. These results suggest that overexpression of the human MrgX3 gene causes a disturbance of the normal cell-differentiation process.

  19. Periodic patterning stem cells and induction of skin appendages: p-ERK-dependent mes-enchymal condensation is coupled with Turing mechanism to convert stripes to spots

    E-Print Network [OSTI]

    Maini, Philip K.

    ABSTRACTS 931 Periodic patterning stem cells and induction of skin appendages: p-ERK-dependent mes patterns remains unknown. Using the feather model, here we show ERK activity-dependent mesenchymal cell chemotaxis toward initial peaks is essential for completing pattern formation. Adding ERK inhibitors produced

  20. University of Connecticut Stem Cell Core Registration Application for Human Embryonic Stem Cell Culture Training

    E-Print Network [OSTI]

    University of Connecticut Stem Cell Core Registration Application for Human Embryonic Stem Cell: (required) Date: #12;University of Connecticut Stem Cell Core Registration Application for Human Embryonic copy to: Internal Mail: MC-3301 OR Mail: UConn Stem Cell Core University of Connecticut Health Center

  1. The role of let-7 in human embryonic stem cell-derived neural precursor cells

    E-Print Network [OSTI]

    Chen, Connie

    2012-01-01

    L. (2010). Neural crest-derived stem cells. Stembook,A.V. (2011). Human ESC-derived neural crest model reveals ahuman embryonic stem cell-derived motoneurons. Stem Cell 25:

  2. Stress-induced changes in gene interactions in human cells

    E-Print Network [OSTI]

    Ives, Zachary G.

    Stress-induced changes in gene interactions in human cells Renuka R. Nayak1 , William E. Bernal2 environments by changing gene expression and gene interactions. To study how human cells response to stress, we endo- plasmic reticulum stress and exposure to ionizing radiation. We identified thousands of genes

  3. Continuous human cell lines and method of making same

    DOE Patents [OSTI]

    Stampfer, M.R.

    1985-07-01

    Substantially genetically stable continuous human cell lines derived from normal human mammary epithelial cells (HMEC) and processes for making and using the same. In a preferred embodiment, the cell lines are derived by treating normal human mammary epithelial tissue with a chemical carcinogen such as benzo(a)pyrene. The novel cell lines serve as useful substrates for elucidating the potential effects of a number of toxins, carcinogens and mutagens as well as of the addition of exogenous genetic material. The autogenic parent cells from which the cell lines are derived serve as convenient control samples for testing. The cell lines are not neoplastically transformed, although they have acquired several properties which distinguish them from their normal progenitors. 2 tabs.

  4. Skin flicks

    E-Print Network [OSTI]

    Orth, Margaret A. (Margaret Ann), 1964-

    1993-01-01

    The written and artistic part of this thesis are both separated into the two categories of "SKIN" and "FLICKS". The Artistic part of my thesis consists of five artificial skins made on my body, and a series of video tapes ...

  5. Comparison of blood flow and cell function in ischemic skin flaps

    SciTech Connect (OSTI)

    Bean, D.; Rees, R.S.; O'Leary, J.P.; Lynch, J.B.

    1984-07-01

    Cellular function and blood flow in acute, steroid-treated, and surgically delayed random skin flaps have been examined. In these studies, the period following flap elevation could be divided into early (0-2 hr), intermediate (4-6 hr), and late (12 hr) periods of ischemia, based on the cutaneous blood flow and cellular function measured by thallium-201 uptake. There was a close correlation between blood flow and cellular function during the early period of ischemia which became worse with time. Blood flow studies demonstrated a significant difference between the early and intermediate periods of ischemia which was abolished by surgical delay. Improvement in cellular function was accomplished by improved blood flow in the surgically delayed flaps, while steroid-treated flaps enhanced cellular metabolism by another mechanism. Cellular function approximated blood flow during the early and immediate period of ischemia. Steroids may augment cellular function without improving blood flow, while surgical delay improves cellular function by improving blood flow.

  6. Wild-type human p53 transactivates the human proliferating cell nuclear antigen promoter

    SciTech Connect (OSTI)

    Shivakumar, C.V.; Brown, D.R.; Deb, S.; Deb, S.P. [Univ. of Texas Health Science Center, San Antonio, TX (United States)

    1995-12-01

    The p53 tumor suppressor protein negatively regulates cell growth and somatic mutations in the p53 gene lead to uncontrolled cell growth and oncogenesis. This report describes research which demonstrates, using a number of different cell lines, that at low levels, wild-type p53 transactivates the human proliferating cell nuclear antigen (PCNA) promoter. When expressed at similar levels, tumor-derived p53 mutants did not transactivate the PCNA promoter. It also reports the identification of a wild-type human p53-binding site on the human PCNA promote. 84 refs., 5 figs, 3 tabs.

  7. Human papillomavirus 16 E5 induces bi-nucleated cell formation by cell-cell fusion

    SciTech Connect (OSTI)

    Hu Lulin; Plafker, Kendra [Department of Cell Biology, University of Oklahoma (United States); Vorozhko, Valeriya [Department of Cell Biology, University of Oklahoma (United States); Cell Cycle and Cancer Biology Research Program, Oklahoma Medical Research Foundation (United States); Zuna, Rosemary E. [Department of Pathology, University of Oklahoma HSC (United States); Hanigan, Marie H. [Department of Cell Biology, University of Oklahoma (United States); Gorbsky, Gary J. [Department of Cell Biology, University of Oklahoma (United States); Cell Cycle and Cancer Biology Research Program, Oklahoma Medical Research Foundation (United States); Plafker, Scott M. [Department of Cell Biology, University of Oklahoma (United States); Angeletti, Peter C. [Nebraska Center for Virology (United States); Ceresa, Brian P. [Department of Cell Biology, University of Oklahoma (United States)], E-mail: brian-ceresa@oushc.edu

    2009-02-05

    Human papillomaviruses (HPV) 16 is a DNA virus encoding three oncogenes - E5, E6, and E7. The E6 and E7 proteins have well-established roles as inhibitors of tumor suppression, but the contribution of E5 to malignant transformation is controversial. Using spontaneously immortalized human keratinocytes (HaCaT cells), we demonstrate that expression of HPV16 E5 is necessary and sufficient for the formation of bi-nucleated cells, a common characteristic of precancerous cervical lesions. Expression of E5 from non-carcinogenic HPV6b does not produce bi-nucleate cells. Video microscopy and biochemical analyses reveal that bi-nucleates arise through cell-cell fusion. Although most E5-induced bi-nucleates fail to propagate, co-expression of HPV16 E6/E7 enhances the proliferation of these cells. Expression of HPV16 E6/E7 also increases bi-nucleated cell colony formation. These findings identify a new role for HPV16 E5 and support a model in which complementary roles of the HPV16 oncogenes lead to the induction of carcinogenesis.

  8. Robust derivation of epicardium and its differentiated smooth muscle cell progeny from human pluripotent stem cells

    E-Print Network [OSTI]

    Iyer, Dharini; Gambardella, Laure; Bernard, William G.; Serrano, Felipe; Mascetti, Victoria L.; Pedersen, Roger A.; Talasila, Amarnath; Sinha, Sanjay

    2015-03-26

    and its potential ability to initiate myocardial repair in injured adult tissues. Here, we describe a chemically defined method for generating epicardium and epicardium-derived smooth muscle cells (EPI-SMCs) and CFs from human pluripotent stem cells (HPSCs...

  9. including cell-cycle regulation, and, along with hTERT, can immortalize human cells6

    E-Print Network [OSTI]

    Balibar, Sébastien

    including cell-cycle regulation, and, along with hTERT, can immortalize human cells6 . So it is possible that a central effect of these two pro- teins is to activate the cell-division machinery in chromosomes), thereby facilitating reprogramming. Are the iPS cells that Park et al. generated, or indeed

  10. Expansion of Human Cord Blood Hematopoietic Stem Cells for Transplantation

    E-Print Network [OSTI]

    Chou, Song

    A recent Science paper reported a purine derivative that expands human cord blood hematopoietic stem cells in culture ( Boitano et al., 2010) by antagonizing the aryl hydrocarbon receptor. Major problems need to be overcome ...

  11. The Effect of TNF- alpha On The Odontogenic Potential Of Human Dental Stem Cells

    E-Print Network [OSTI]

    Tseng, Edward

    2012-01-01

    dental  pulp  stem  cells  (hDPSCs)  and  human   mesenchymal  stem  cells  (hMSCs)  for  tissue  engineering  cell  therapy.  Biomaterials,  

  12. Regulation of apoptosis in human cancer cells

    E-Print Network [OSTI]

    Lloyd, S. Julie-Ann (Simone Julie-Ann)

    2005-01-01

    Nitric oxide is postulated to protect cancer cells from the death-inducing effects of tumour necrosis factor alpha by S-nitrosating the active site cysteines, inhibiting cleavage of caspase-9. We aimed to test this hypothesis ...

  13. Distribution of mutant cells in human skin : exploration of the fetal-juvenile mutability hypothesis

    E-Print Network [OSTI]

    Kao, Leslie E

    2009-01-01

    The multiple "hits" carcinogenesis models are extensions of the cancer incidence theory developed by researchers from Nordling (1953), Armitage-Doll (1954 and 1957), Knudson (1971), Moolgavkar and Verzon (1979), to Moolgavkar ...

  14. Teroxirone inhibited growth of human non-small cell lung cancer cells by activating p53

    SciTech Connect (OSTI)

    Wang, Jing-Ping; Lin, Kai-Han; Liu, Chun-Yen; Yu, Ya-Chu; Wu, Pei-Tsun [Department of Life Science, National Taiwan Normal University, Taipei, Taiwan (China); Chiu, Chien-Chih [Department of Biotechnology, Kaohsiung Medical University, Kaohsiung, Taiwan (China); Su, Chun-Li [Department of Human Development and Family Studies, National Taiwan Normal University, Taipei, Taiwan (China); Chen, Kwun-Min [Department of Chemistry, National Taiwan Normal University, Taipei, Taiwan (China); Fang, Kang, E-mail: kangfang@ntnu.edu.tw [Department of Life Science, National Taiwan Normal University, Taipei, Taiwan (China)

    2013-11-15

    In this work, we demonstrated that the growth of human non-small-cell-lung-cancer cells H460 and A549 cells can be inhibited by low concentrations of an epoxide derivative, teroxirone, in both in vitro and in vivo models. The cytotoxicity was mediated by apoptotic cell death through DNA damage. The onset of ultimate apoptosis is dependent on the status of p53. Teroxirone caused transient elevation of p53 that activates downstream p21 and procaspase-3 cleavage. The presence of caspase-3 inhibitor reverted apoptotic phenotype. Furthermore, we showed the cytotoxicity of teroxirone in H1299 cells with stable ectopic expression of p53, but not those of mutant p53. A siRNA-mediated knockdown of p53 expression attenuated drug sensitivity. The in vivo experiments demonstrated that teroxirone suppressed growth of xenograft tumors in nude mice. Being a potential therapeutic agent by restraining cell growth through apoptotic death at low concentrations, teroxirone provides a feasible perspective in reversing tumorigenic phenotype of human lung cancer cells. - Highlights: • Teroxirone repressed tumor cell growth in nude mice of human lung cancer cells. • The apoptotic cell death reverted by caspase-3 inhibitor is related to p53 status. • Teroxirone provides a good candidate for lung cancer treatment.

  15. Cell surface display of functional human MHC class II proteins: yeast display versus insect cell display

    E-Print Network [OSTI]

    Zhao, Huimin

    Cell surface display of functional human MHC class II proteins: yeast display versus insect cell histocompatibility complex class II (MHCII) pro- teins have proved elusive. Availability of such systems would enable success, the tetramer technology has been more difficult to use for MHC class II molecules, mainly due

  16. Healthy Skin Matters Normal Skin

    E-Print Network [OSTI]

    Rau, Don C.

    into contact with the rest of the world. It holds body fluids in, preventing dehydration (dee when your skin is injured If you do get a cut or scratch, clean it right away with soap and warm water

  17. Trichodermin induces cell apoptosis through mitochondrial dysfunction and endoplasmic reticulum stress in human chondrosarcoma cells

    SciTech Connect (OSTI)

    Su, Chen-Ming; Wang, Shih-Wei; Lee, Tzong-Huei; Tzeng, Wen-Pei; Hsiao, Che-Jen; Liu, Shih-Chia; Tang, Chih-Hsin

    2013-10-15

    Chondrosarcoma is the second most common primary bone tumor, and it responds poorly to both chemotherapy and radiation treatment. Nalanthamala psidii was described originally as Myxosporium in 1926. This is the first study to investigate the anti-tumor activity of trichodermin (trichothec-9-en-4-ol, 12,13-epoxy-, acetate), an endophytic fungal metabolite from N. psidii against human chondrosarcoma cells. We demonstrated that trichodermin induced cell apoptosis in human chondrosarcoma cell lines (JJ012 and SW1353 cells) instead of primary chondrocytes. In addition, trichodermin triggered endoplasmic reticulum (ER) stress protein levels of IRE1, p-PERK, GRP78, and GRP94, which were characterized by changes in cytosolic calcium levels. Furthermore, trichodermin induced the upregulation of Bax and Bid, the downregulation of Bcl-2, and the dysfunction of mitochondria, which released cytochrome c and activated caspase-3 in human chondrosarcoma. In addition, animal experiments illustrated reduced tumor volume, which led to an increased number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells and an increased level of cleaved PARP protein following trichodermin treatment. Together, this study demonstrates that trichodermin is a novel anti-tumor agent against human chondrosarcoma cells both in vitro and in vivo via mitochondrial dysfunction and ER stress. - Highlights: • Trichodermin induces chondrosarcoma apoptosis. • ER stress is involved in trichodermin-induced cell death. • Trichodermin induces chondrosarcoma death in vivo.

  18. Raman Spectroscopy of DNA Packaging in Individual Human Sperm Cells distinguishes Normal from Abnormal Cells

    SciTech Connect (OSTI)

    Huser, T; Orme, C; Hollars, C; Corzett, M; Balhorn, R

    2009-03-09

    Healthy human males produce sperm cells of which about 25-40% have abnormal head shapes. Increases in the percentage of sperm exhibiting aberrant sperm head morphologies have been correlated with male infertility, and biochemical studies of pooled sperm have suggested that sperm with abnormal shape may contain DNA that has not been properly repackaged by protamine during spermatid development. We have used micro-Raman spectroscopy to obtain Raman spectra from individual human sperm cells and examined how differences in the Raman spectra of sperm chromatin correlate with cell shape. We show that Raman spectra of individual sperm cells contain vibrational marker modes that can be used to assess the efficiency of DNA-packaging for each cell. Raman spectra obtained from sperm cells with normal shape provide evidence that DNA in these sperm is very efficiently packaged. We find, however, that the relative protein content per cell and DNA packaging efficiencies are distributed over a relatively wide range for sperm cells with both normal and abnormal shape. These findings indicate that single cell Raman spectroscopy should be a valuable tool in assessing the quality of sperm cells for in-vitro fertilization.

  19. Somatic cell genotoxicity at the glycophorin A locus in humans

    SciTech Connect (OSTI)

    Jensen, R.H.; Grant, S.G.; Langlois, R.G.; Bigbee, W.L.

    1990-12-28

    We have developed an assay for detecting variant erythrocytes that occur as a result of in vivo allele loss at the glycophorin A (GPA) locus on chromosome 4 in humans. This gene codes for an erythroid- specific cell surface glycoprotein, and with our assay we are able to detect rare variant erythrocytes that have lost expression of one of the two GPA alleles. Two distinctly different variant cell types are detected with this assay. One variant cell type (called N{O}) is hemizygous. Our assay also detects homozygous variant erythrocytes that have lost expression of the GPA(M) allele and express the GPA(N) allele at twice the heterozygous level. The results of this assay are an enumeration of the frequency of N{O} and NN variant cell types for each individual analyzed. These variant cell frequencies provide a measure of the amount of somatic cell genotoxicity that has occurred at the GPA locus. Such genotoxicity could be the result of (1) reactions of toxic chemicals to which the individual has been exposed, or (2) high energy radiation effects on erythroid precursor cells, or (3) errors in DNA replication or repair in these cells of the bone marrow. Thus, the GPA-based variant cell frequency can serve as a biodosimeter that indicates the amount of genotoxic exposure each individual has received. Because two very different kinds of variant cells are enumerated, different kinds of genotoxicity should be distinguishable. Results of the GPA somatic genotoxicity assay may also provide valuable information for cancer-risk estimation on each individual. 16 refs.

  20. In vivo imaging of immune cell dynamics in skin in response to zinc-oxide nanoparticle exposure

    E-Print Network [OSTI]

    Boppart, Stephen

    in cosmetic and sunscreen products which are applied topically to the skin. Despite their widespread use of nanosized particles in titanium dioxide- and zinc oxide-based sunscreens," J. Am. Acad. Dermatol. 61(4), 685 by cosmetic pigments and sunscreen agents under solar exposure and artificial UV illumination," J. Oleo Sci

  1. Fulvestrant radiosensitizes human estrogen receptor-positive breast cancer cells

    SciTech Connect (OSTI)

    Wang, Jing; Department of Oncology, Affiliated Hospital of Qingdao University Medical College, Shandong Province ; Yang, Qifeng; Haffty, Bruce G.; Li, Xiaoyan; Moran, Meena S.

    2013-02-08

    Highlights: ? Fulvestrant radiosensitizes MCF-7 cells. ? Fulvestrant increases G1 arrest and decreases S phase in MCF-7 cells. ? Fulvestrant down-regulates DNA-PKcs and RAD51 in MCF-7 cells. -- Abstract: The optimal sequencing for hormonal therapy and radiation are yet to be determined. We utilized fulvestrant, which is showing promise as an alternative to other agents in the clinical setting of hormonal therapy, to assess the cellular effects of concomitant anti-estrogen therapy (fulvestrant) with radiation (F + RT). This study was conducted to assess the effects of fulvestrant alone vs. F + RT on hormone-receptor positive breast cancer to determine if any positive or negative combined effects exist. The effects of F + RT on human breast cancer cells were assessed using MCF-7 clonogenic and tetrazolium salt colorimetric (MTT) assays. The assays were irradiated with a dose of 0, 2, 4, 6 Gy ± fulvestrant. The effects of F + RT vs. single adjuvant treatment alone on cell-cycle distribution were assessed using flow cytometry; relative expression of repair proteins (Ku70, Ku80, DNA-PKcs, Rad51) was assessed using Western Blot analysis. Cell growth for radiation alone vs. F + RT was 0.885 ± 0.013 vs. 0.622 ± 0.029 @2 Gy, 0.599 ± 0.045 vs. 0.475 ± 0.054 @4 Gy, and 0.472 ± 0.021 vs. 0.380 ± 0.018 @6 Gy RT (p = 0.003). While irradiation alone induced G2/M cell cycle arrest, the combination of F + RT induced cell redistribution in the G1 phase and produced a significant decrease in the proportion of cells in G2 phase arrest and in the S phase in breast cancer cells (p < 0.01). Furthermore, levels of repair proteins DNA-PKcs and Rad51 were significantly decreased in the cells treated with F + RT compared with irradiation alone. F + RT leads to a decrease in the surviving fraction, increased cell cycle arrest, down regulating of nonhomologous repair protein DNA-PKcs and homologous recombination repair protein RAD51. Thus, our findings suggest that F + RT increases breast cancer cell radiosensitivity compared with radiation alone. These findings have salient implications for designing clinical trials using fulvestrant and radiation therapy.

  2. Urocortin 3 Marks Mature Human Primary and Embryonic Stem Cell-Derived Pancreatic Alpha and Beta

    E-Print Network [OSTI]

    Sander, Maike

    Urocortin 3 Marks Mature Human Primary and Embryonic Stem Cell-Derived Pancreatic Alpha and Beta cells. To substantiate these findings, we analyzed human embryonic stem cell (hESC)-derived pancreatic endoderm that differentiates into mature endocrine cells upon engraftment in mice. Ucn 3 expression in hESC-derived

  3. PROTOCOLS FOR HUMAN EMBRYONIC STEM CELL WORK THE BRIVANLOU LABORATORY OF MOLECULAR EMBRYOLOGY

    E-Print Network [OSTI]

    de Lange, Titia

    ) and inactivated using Mitomycin-C or gamma irradiation or commercially available Mitomycin-C treated MEFs scissors make a cut across the belly and cut away the skin to expose the gut. With sterile forceps in a total of 30 mls of medium per flask. This density allows the cells to adhere but not become overly

  4. Selective Destruction Of Cells Infected With The Human Immunodeficiency Virus

    DOE Patents [OSTI]

    Keener, William K. (Idaho Falls, ID); Ward, Thomas E. (Idaho Falls, ID)

    2006-03-28

    Compositions and methods for selectively killing a cell containing a viral protease are disclosed. The composition is a varient of a protein synthesis inactivating toxin wherein a viral protease cleavage site is interposed between the A and B chains. The variant of the type II ribosome-inactivating protein is activated by digestion of the viral protease cleavage site by the specific viral protease. The activated ribosome-inactivating protein then kills the cell by inactivating cellular ribosomes. A preferred embodiment of the invention is specific for human immunodeficiency virus (HIV) and uses ricin as the ribosome-inactivating protein. In another preferred embodiment of the invention, the variant of the ribosome-inactivating protein is modified by attachment of one or more hydrophobic agents. The hydrophobic agent facilitates entry of the variant of the ribosome-inactivating protein into cells and can lead to incorporation of the ribosome-inactivating protein into viral particles. Still another preferred embodiment of the invention includes a targeting moiety attached to the variants of the ribosome-inactivating protein to target the agent to HIV infectable cells.

  5. Selective destruction of cells infected with human immunodeficiency virus

    DOE Patents [OSTI]

    Keener, William K.; Ward, Thomas E.

    2003-09-30

    Compositions and methods for selectively killing a cell containing a viral protease are disclosed. The composition is a variant of a protein synthesis inactivating toxin wherein a viral protease cleavage site is interposed between the A and B chains. The variant of the type II ribosome-inactivating protein is activated by digestion of the viral protease cleavage site by the specific viral protease. The activated ribosome-inactivating protein then kills the cell by inactivating cellular ribosomes. A preferred embodiment of the invention is specific for human immunodeficiency virus (HIV) and uses ricin as the ribosome-inactivating protein. In another preferred embodiment of the invention, the variant of the ribosome-inactivating protein is modified by attachment of one or more hydrophobic agents. The hydrophobic agent facilitates entry of the variant of the ribosome-inactivating protein into cells and can lead to incorporation of the ribosome-inactivating protein into viral particles. Still another preferred embodiment of the invention includes a targeting moiety attached to the variants of the ribosome-inactivating protein to target the agent to HIV infectable cells.

  6. Mesenchymal stem cells secreting angiopoietin-like-5 support efficient expansion of human hematopoietic stem cells without compromising their repopulating potential

    E-Print Network [OSTI]

    Drake, Adam

    Clinical and preclinical applications of human hematopoietic stem cells (HSCs) are often limited by scarcity of cells. Expanding human HSCs to increase their numbers while maintaining their stem cell properties has therefore ...

  7. Vorinostat, an HDAC inhibitor attenuates epidermoid squamous cell carcinoma growth by dampening mTOR signaling pathway in a human xenograft murine model

    SciTech Connect (OSTI)

    Kurundkar, Deepali; Srivastava, Ritesh K.; Chaudhary, Sandeep C. [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, VH 509, Birmingham, AL 35294-0019 (United States)] [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, VH 509, Birmingham, AL 35294-0019 (United States); Ballestas, Mary E. [Department of Pediatrics Infectious Disease, Children's of Alabama, School of Medicine, University of Alabama at Birmingham, AL (United States)] [Department of Pediatrics Infectious Disease, Children's of Alabama, School of Medicine, University of Alabama at Birmingham, AL (United States); Kopelovich, Levy [Division of Cancer Prevention, National Cancer Institute, 6130 Executive Blvd., Suite 2114, Bethesda, MD 20892 (United States)] [Division of Cancer Prevention, National Cancer Institute, 6130 Executive Blvd., Suite 2114, Bethesda, MD 20892 (United States); Elmets, Craig A. [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, VH 509, Birmingham, AL 35294-0019 (United States)] [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, VH 509, Birmingham, AL 35294-0019 (United States); Athar, Mohammad, E-mail: mathar@uab.edu [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, VH 509, Birmingham, AL 35294-0019 (United States)] [Department of Dermatology and Skin Diseases Research Center, University of Alabama at Birmingham, 1530 3rd Avenue South, VH 509, Birmingham, AL 35294-0019 (United States)

    2013-01-15

    Histone deacetylase (HDAC) inhibitors are potent anticancer agents and show efficacy against various human neoplasms. Vorinostat is a potent HDAC inhibitor and has shown potential to inhibit growth of human xenograft tumors. However, its effect on the growth of skin neoplasm remains undefined. In this study, we show that vorinostat (2 ?M) reduced expression of HDAC1, 2, 3, and 7 in epidermoid carcinoma A431 cells. Consistently, it increased acetylation of histone H3 and p53. Vorinostat (100 mg/kg body weight, IP) treatment reduced human xenograft tumor growth in highly immunosuppressed nu/nu mice. Histologically, the vorinostat-treated tumor showed features of well-differentiation with large necrotic areas. Based on proliferating cell nuclear antigen (PCNA) staining and expression of cyclins D1, D2, E, and A, vorinostat seems to impair proliferation by down-regulating the expression of these proteins. However, it also induced apoptosis. The mechanism by which vorinostat blocks proliferation and makes tumor cells prone to apoptosis, involved inhibition of mTOR signaling which was accompanied by reduction in cell survival AKT and extracellular-signal regulated kinase (ERK) signaling pathways. Our data provide a novel mechanism-based therapeutic intervention for cutaneous squamous cell carcinoma (SCC). Vorinostat may be utilized to cure skin neoplasms in organ transplant recipient (OTR). These patients have high morbidity and surgical removal of these lesions which frequently develop in these patients, is difficult. -- Highlights: ? Vorinostat reduces SCC growth in a xenograft murine model. ? Vorinostat dampens proliferation and induces apoptosis in tumor cells. ? Diminution in mTOR, Akt and ERK signaling underlies inhibition in proliferation. ? Vorinostat by inhibiting HDACs inhibits epithelial–mesenchymal transition.

  8. The Skin Microbiome in Healthy and Allergic Dogs 

    E-Print Network [OSTI]

    Hoffmann, Aline Rodrigues; Patterson, Adam P.; Diesel, Alison; Lawhon, Sara D.; Ly, Hoai Jaclyn; Stephenson, Christine Elkins; Mansell, Joanne; Steiner, Jö rg M.; Dowd, Scot E.; Olivry, Thierry; Suchodolski, Jan S.

    2014-01-08

    Changes in the microbial populations on the skin of animals have traditionally been evaluated using conventional microbiology techniques. The sequencing of bacterial 16S rRNA genes has revealed that the human skin is inhabited by a highly diverse...

  9. The transcriptome of human CD34 hematopoietic stem-progenitor cells

    E-Print Network [OSTI]

    cells, including the de novo-generated 3 ESTs and the existing sequences of full-length cDNAs, ESTsThe transcriptome of human CD34 hematopoietic stem-progenitor cells Yeong C. Kima,1 , Qingfa Wua,1 for understanding the genetic basis of hematopoiesis. We analyzed gene expression in human CD34 hematopoietic cells

  10. Design and Analysis of a Wireless Nanosensor Network for Monitoring Human Lung Cells

    E-Print Network [OSTI]

    New South Wales, University of

    Design and Analysis of a Wireless Nanosensor Network for Monitoring Human Lung Cells Eisa Zarepour one day. In this paper, we design and analyse a WNSN for monitoring human lung cells. We find that influences the terahertz channel inside lung cells. The channel is characterised as a two-state channel

  11. pH-sensitive intracellular photoluminescence of carbon nanotubefluorescein conjugates in human ovarian cancer cells

    E-Print Network [OSTI]

    Zhou, Chongwu

    ovarian cancer cells This article has been downloaded from IOPscience. Please scroll down to see the full photoluminescence of carbon nanotube­fluorescein conjugates in human ovarian cancer cells M T Chen1 , L M Gomez2 , F-walled carbon nanotube­fluorescein carbazide (SWCNT­FC) conjugates in human ovarian cancer cells. Light

  12. Role of copper in the regulation of CU, ZN-superoxide dismutase in human K562 erythroleukemia cells and human fibroblasts 

    E-Print Network [OSTI]

    Yu, Dan

    1994-01-01

    Activation of the enzyme CU2Zn2-SUperoxide dismutase (CuZnSOD) by its copper cofactor was studied in K562 erythroleukemia cells and skin fibroblasts. K562 cells were incubated in medium supplemented with 0-50 IIM CUC12 or ZnCI2 for 24 h and extracts...

  13. Induction of apoptotic death and retardation of neuronal differentiation of human neural stem cells by sodium arsenite treatment

    SciTech Connect (OSTI)

    Ivanov, Vladimir N., E-mail: vni3@columbia.edu [Center for Radiological Research, Department of Radiation Oncology, College of Physicians and Surgeons, Columbia University, 630 West 168th Street, NY 10032 (United States); Hei, Tom K. [Center for Radiological Research, Department of Radiation Oncology, College of Physicians and Surgeons, Columbia University, 630 West 168th Street, NY 10032 (United States)

    2013-04-01

    Chronic arsenic toxicity is a global health problem that affects more than 100 million people worldwide. Long-term health effects of inorganic sodium arsenite in drinking water may result in skin, lung and liver cancers and in severe neurological abnormalities. We investigated in the present study whether sodium arsenite affects signaling pathways that control cell survival, proliferation and neuronal differentiation of human neural stem cells (NSC). We demonstrated that the critical signaling pathway, which was suppressed by sodium arsenite in NSC, was the protective PI3K–AKT pathway. Sodium arsenite (2–4 ?M) also caused down-regulation of Nanog, one of the key transcription factors that control pluripotency and self-renewal of stem cells. Mitochondrial damage and cytochrome-c release induced by sodium arsenite exposure was followed by initiation of the mitochondrial apoptotic pathway in NSC. Beside caspase-9 and caspase-3 inhibitors, suppression of JNK activity decreased levels of arsenite-induced apoptosis in NSC. Neuronal differentiation of NSC was substantially inhibited by sodium arsenite exposure. Overactivation of JNK1 and ERK1/2 and down-regulation of PI3K–AKT activity induced by sodium arsenite were critical factors that strongly affected neuronal differentiation. In conclusion, sodium arsenite exposure of human NSC induces the mitochondrial apoptotic pathway, which is substantially accelerated due to the simultaneous suppression of PI3K–AKT. Sodium arsenite also negatively affects neuronal differentiation of NSC through overactivation of MEK–ERK and suppression of PI3K–AKT. - Highlights: ? Arsenite induces the mitochondrial apoptotic pathway in human neural stem cells. ? Arsenite-induced apoptosis is strongly upregulated by suppression of PI3K–AKT. ? Arsenite-induced apoptosis is strongly down-regulated by inhibition of JNK–cJun. ? Arsenite negatively affects neuronal differentiation by inhibition of PI3K–AKT.

  14. The influence of microwave radiation on the state of chromatin in human cells

    E-Print Network [OSTI]

    Y. G. Shckorbatov; V. N. Pasiuga; V. A. Grabina; N. N. Kolchigin; D. O. Batrakov; V. V. Kalashnikov; D. D. Ivanchenko; V. N. Bykov

    2008-09-03

    Isolated human buccal epithelium cell were irradiated by microwaves at frequency f=35 GHz and surface power density E=30 mcW/cm2. The state of chromatin in human cells was determined by methodsof light and electron microscopy. The state of cell membranes was evaluated by the method of vital indigo carmine staining. The microwave-induced condensation of chromatin in human cells was revealed. Left side circulary polarized waves induced less effect than linearly polarized radiation. The linearly polarized electromagnetic waves induced cell membrane damage revealed by the increase of cell stainability. The data obtained are discussed in connection with the mechanisms of biologica effect of electromagnetic waves.

  15. Skin Bleaching in Jamaica: A Colonial Legacy 

    E-Print Network [OSTI]

    Robinson, Petra Alaine

    2012-07-16

    -1 SKIN BLEACHING IN JAMAICA: A COLONIAL LEGACY A Dissertation by PETRA ALAINE ROBINSON Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY... May 2011 Major Subject: Educational Human Resource Development Skin Bleaching in Jamaica: A Colonial Legacy Copyright 2011 Petra Alaine Robinson SKIN BLEACHING IN JAMAICA: A COLONIAL LEGACY...

  16. Molecular Signatures of Human Induced Pluripotent Stem Cells Highlight Sex Differences and Cancer Genes

    E-Print Network [OSTI]

    Anguera, Montserrat C.

    Although human induced pluripotent stem cells (hiPSCs) have enormous potential in regenerative medicine, their epigenetic variability suggests that some lines may not be suitable for human therapy. There are currently few ...

  17. Modeling hepatitis C virus infection using human induced pluripotent stem cells

    E-Print Network [OSTI]

    Schwartz, Robert E.

    Human pathogens impact patient health through a complex interplay with the host, but models to study the role of host genetics in this process are limited. Human induced pluripotent stem cells (iPSCs) offer the ability to ...

  18. Prodigiosin activates endoplasmic reticulum stress cell death pathway in human breast carcinoma cell lines

    SciTech Connect (OSTI)

    Pan, Mu-Yun [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China)] [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Shen, Yuh-Chiang [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China) [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); National Research Institute of Chinese Medicine, Taipei, Taiwan (China); Lu, Chien-Hsing [Department of Obstetrics and Gynecology, Taichung Veterans General Hospital, Taichung, Taiwan (China) [Department of Obstetrics and Gynecology, Taichung Veterans General Hospital, Taichung, Taiwan (China); Department of Obstetrics and Gynecology, National Yang-Ming University School of Medicine, Taipei, Taiwan (China); Yang, Shu-Yi [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China)] [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Ho, Tsing-Fen [Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan (China)] [Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, Taiwan (China); Peng, Yu-Ta [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China)] [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Chang, Chia-Che, E-mail: chia_che@dragon.nchu.edu.tw [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China) [Institute of Biomedical Sciences, National Chung Hsing University, Taichung, Taiwan (China); Agricultural Biotechnology Center, National Chung Hsing University, Taichung, Taiwan (China); Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan (China)

    2012-12-15

    Prodigiosin is a bacterial tripyrrole pigment with potent cytotoxicity against diverse human cancer cell lines. Endoplasmic reticulum (ER) stress is initiated by accumulation of unfolded or misfolded proteins in the ER lumen and may induce cell death when irremediable. In this study, the role of ER stress in prodigiosin-induced cytotoxicity was elucidated for the first time. Comparable to the ER stress inducer thapsigargin, prodigiosin up-regulated signature ER stress markers GRP78 and CHOP in addition to activating the IRE1, PERK and ATF6 branches of the unfolded protein response (UPR) in multiple human breast carcinoma cell lines, confirming prodigiosin as an ER stress inducer. Prodigiosin transcriptionally up-regulated CHOP, as evidenced by its promoting effect on the CHOP promoter activity. Of note, knockdown of CHOP effectively lowered prodigiosin's capacity to evoke PARP cleavage, reduce cell viability and suppress colony formation, highlighting an essential role of CHOP in prodigiosin-induced cytotoxic ER stress response. In addition, prodigiosin down-regulated BCL2 in a CHOP-dependent manner. Importantly, restoration of BCL2 expression blocked prodigiosin-induced PARP cleavage and greatly enhanced the survival of prodigiosin-treated cells, suggesting that CHOP-dependent BCL2 suppression mediates prodigiosin-elicited cell death. Moreover, pharmacological inhibition of JNK by SP600125 or dominant-negative blockade of PERK-mediated eIF2? phosphorylation impaired prodigiosin-induced CHOP up-regulation and PARP cleavage. Collectively, these results identified ER stress-mediated cell death as a mode-of-action of prodigiosin's tumoricidal effect. Mechanistically, prodigiosin engages the IRE1–JNK and PERK–eIF2? branches of the UPR signaling to up-regulate CHOP, which in turn mediates BCL2 suppression to induce cell death. Highlights: ? Prodigiosin is a bacterial tripyrrole pigment with potent anticancer effect. ? Prodigiosin is herein identified as an endoplasmic reticulum (ER) stress inducer. ? Prodigiosin-induced cytotoxicity involves ER stress-mediated cell death. ? Prodigiosin transcriptionally induces CHOP to suppress BCL2 for evoking cell death. ? Prodigiosin engages the IRE1–JNK and PERK–eIF2? pathways to up-regulate CHOP.

  19. Cathepsin S regulates class II MHC processing in human CD4+ HLA-DR+ T cells

    E-Print Network [OSTI]

    Ploegh, Hidde

    Although it has long been known that human CD4+ T cells can express functional class II MHC molecules, the role of lysosomal proteases in the T cell class II MHC processing and presentation pathway is unknown. Using CD4+ ...

  20. Dynamic behavior of healthy and malaria infected human red blood cells

    E-Print Network [OSTI]

    Quinn, David John, Sc. D. Massachusetts Institute of Technology

    2010-01-01

    Hereditary hematological disorders and foreign organisms often introduce changes to the spectrin molecular network and membrane of human red blood cells (RBCs). These structural changes lead to altered cell shape, ...

  1. Combinatorial Development of Biomaterials for Clonal Growth of Human Pluripotent Stem Cells

    E-Print Network [OSTI]

    Mei, Ying

    Both human embryonic stem cells and induced pluripotent stem cells can self-renew indefinitely in culture; however, present methods to clonally grow them are inefficient and poorly defined for genetic manipulation and ...

  2. Equivalence of Conventionally-Derived and Parthenote-Derived Human Embryonic Stem Cells

    E-Print Network [OSTI]

    2011-01-01

    Equivalence of Conventionally-Derived andParthenote- Derived Human Embryonic Stem Cells Julie V.cell (hESC) lines can be derived via multiple means, it is

  3. Predicting human developmental toxicity of pharmaceuticals using human embryonic stem cells and metabolomics

    SciTech Connect (OSTI)

    West, Paul R., E-mail: pwest@stemina.co [Stemina Biomarker Discovery, Inc., 504 S. Rosa Rd., Suite 150, Madison, WI 53719 (United States); Weir, April M.; Smith, Alan M.; Donley, Elizabeth L.R. [Stemina Biomarker Discovery, Inc., 504 S. Rosa Rd., Suite 150, Madison, WI 53719 (United States); Cezar, Gabriela G. [Stemina Biomarker Discovery, Inc., 504 S. Rosa Rd., Suite 150, Madison, WI 53719 (United States); University of Wisconsin-Madison, Department of Animal Sciences, 1675 Observatory Drive, Madison, WI 53706 (United States)

    2010-08-15

    Teratogens, substances that may cause fetal abnormalities during development, are responsible for a significant number of birth defects. Animal models used to predict teratogenicity often do not faithfully correlate to human response. Here, we seek to develop a more predictive developmental toxicity model based on an in vitro method that utilizes both human embryonic stem (hES) cells and metabolomics to discover biomarkers of developmental toxicity. We developed a method where hES cells were dosed with several drugs of known teratogenicity then LC-MS analysis was performed to measure changes in abundance levels of small molecules in response to drug dosing. Statistical analysis was employed to select for specific mass features that can provide a prediction of the developmental toxicity of a substance. These molecules can serve as biomarkers of developmental toxicity, leading to better prediction of teratogenicity. In particular, our work shows a correlation between teratogenicity and changes of greater than 10% in the ratio of arginine to asymmetric dimethylarginine levels. In addition, this study resulted in the establishment of a predictive model based on the most informative mass features. This model was subsequently tested for its predictive accuracy in two blinded studies using eight drugs of known teratogenicity, where it correctly predicted the teratogenicity for seven of the eight drugs. Thus, our initial data shows that this platform is a robust alternative to animal and other in vitro models for the prediction of the developmental toxicity of chemicals that may also provide invaluable information about the underlying biochemical pathways.

  4. RNA-programmed genome editing in human cells Martin Jinek1

    E-Print Network [OSTI]

    Doudna, Jennifer A.

    1 RNA-programmed genome editing in human cells Martin Jinek1 , Alexandra East2 , Aaron endonuclease stimulates site-specific genome editing in human cells. #12; 2 Abstract Type II at the 3' end enhances DNA targeting activity in vivo. These results show that RNA-programmed genome

  5. Stress-specific signatures: expression profiling of p53 wild-type and -null human cells

    E-Print Network [OSTI]

    Stress-specific signatures: expression profiling of p53 wild-type and -null human cells Sally responses of human cell lines exposed to a diverse set of stress agents were compared by cDNA microarray expression signatures were defined that discriminated between four broad general mechanisms of stress agents

  6. Characterization of axon guidance cue sensitivity of human embryonic stem cell-derived dopaminergic neurons

    E-Print Network [OSTI]

    McConnell, Susan

    Characterization of axon guidance cue sensitivity of human embryonic stem cell-derived dopaminergic derived from human embryonic stem cells will be useful in future transplantation studies of Parkinson of hESC-derived neurons to respond to axon guidance cues will be critical. Both Netrin-1 and Slit-2

  7. Chemoprevention of human skin cancers.

    E-Print Network [OSTI]

    Loescher, L J; Meyskens, F L Jr

    1991-01-01

    activity of celecoxib, a specific cyclooxygenase- 2ad- ministration of celecoxib, a selective COX-2 inhibitor,

  8. Investigation of a suppression of asymmetric cell kinetics (SACK) approach for ex vivo expansion of human hematopoietic stem cells

    E-Print Network [OSTI]

    Taghizadeh, Rouzbeh R

    2006-01-01

    Ex vivo expansion of hematopoietic stem cells (HSCs) is a long-standing challenge faced by both researchers and clinicians. To date, no robust, efficient method for the pure, ex vivo expansion of human HSCs has been ...

  9. Characterisation of epithelial progenitor cells for human and mouse thymus 

    E-Print Network [OSTI]

    Farley, Alison

    2009-01-01

    The thymus is a complex cellular structure made up of several interdependent cell types and is the primary site for T cell development. A population of fetal thymic epithelial cells (TEC), marked by MTS20 and MTS24, when ...

  10. Chromosomal changes in cultured human epithelial cells transformed by low- and high-LET radiation

    SciTech Connect (OSTI)

    Yang, Tracy Chui-hsu; Craise, L.M; Prioleau, J.C.; Stampfer, M.R.; Rhim, J.S.

    1990-11-01

    For a better assessment of radiation risk in space, an understanding of the responses of human cells, especially the epithelial cells, to low- and high-LET radiation is essential. In our laboratory, we have successfully developed techniques to study the neoplastic transformation of two human epithelial cell systems by ionizing radiation. These cell systems are human mammary epithelial cells (H184B5) and human epidermal keratinocytes (HEK). Both cell lines are immortal, anchorage dependent for growth, and nontumorigenic in athymic nude nice. Neoplastic transformation was achieved by irradiation cells successively. Our results showed that radiogenic cell transformation is a multistep process and that a single exposure of ionizing radiation can cause only one step of transformation. It requires, therefore, multihits to make human epithelial cells fully tumorigenic. Using a simple karyotyping method, we did chromosome analysis with cells cloned at various stages of transformation. We found no consistent large terminal deletion of chromosomes in radiation-induced transformants. Some changes of total number of chromosomes, however, were observed in the transformed cells. These transformants provide an unique opportunity for further genetic studies at a molecular level. 15 refs., 9 figs., 2 tabs.

  11. Reprogramming of murine and human somatic cells using a single polycistronic vector

    E-Print Network [OSTI]

    Saha, Krishanu

    are equivalent to embryonic stem (ES) cells maintaining a full capacity for differentiation with the abilityReprogramming of murine and human somatic cells using a single polycistronic vector Bryce W. Careya, 2008) Directed reprogramming of somatic cells by defined factors pro- vides a novel method

  12. A microfluidic processor for gene expression profiling of single human embryonic stem cells

    E-Print Network [OSTI]

    Quake, Stephen R.

    A microfluidic processor for gene expression profiling of single human embryonic stem cells Jiang F and differentiation of individual cells. Here, we report a microfluidic approach that can extract total mRNA from. This feature makes large-scale single-cell gene expression profiling possible. Using this microfluidic device

  13. Reflective Terahertz Imaging for early diagnosis of skin burn severity

    E-Print Network [OSTI]

    TEWARI, PRIYAMVADA

    2013-01-01

    regions of skin, burns, psoriasis, basal cell carcinomas andinto diseased states like psoriasis, eczema, basal celland cancer, conditions like psoriasis, eczema, dermatitis,

  14. Lumican induces human corneal epithelial cell migration and integrin expression via ERK 1/2 signaling

    SciTech Connect (OSTI)

    Seomun, Young [Laboratory of Ophthalmology and Visual Science, Korean Eye Tissue and Gene Bank related to Blindness, College of Medicine, Catholic University of Korea, 505 Banpo-dong, Seocho-ku, Seoul 137-040 (Korea, Republic of); Joo, Choun-Ki [Laboratory of Ophthalmology and Visual Science, Korean Eye Tissue and Gene Bank related to Blindness, College of Medicine, Catholic University of Korea, 505 Banpo-dong, Seocho-ku, Seoul 137-040 (Korea, Republic of)], E-mail: ckjoo@catholic.ac.kr

    2008-07-18

    Lumican is a major proteoglycans of the human cornea. Lumican knock-out mice have been shown to lose corneal transparency and to display delayed wound healing. The purpose of this study was to define the role of lumican in corneal epithelial cell migration. Over-expression of lumican in human corneal epithelial (HCE-T) cells increased both cell migration and proliferation, and increased levels of integrins {alpha}2 and {beta}1. ERK 1/2 was also activated in lumican over-expressed cells. When we treated HCE-T cells with the ERK-specific inhibitor U0126, cell migration and the expression of integrin {beta}1 were completely blocked. These data provide evidence that lumican stimulates cell migration in the corneal epithelium by activating ERK 1/2, and point to a novel signaling pathway implicated in corneal epithelial cell migration.

  15. Mortalin antibody-conjugated quantum dot transfer from human mesenchymal stromal cells to breast cancer cells requires cell–cell interaction

    SciTech Connect (OSTI)

    Pietilä, Mika; Lehenkari, Petri; Kuvaja, Paula; Kaakinen, Mika; Kaul, Sunil C.; Wadhwa, Renu; Uemura, Toshimasa

    2013-11-01

    The role of tumor stroma in regulation of breast cancer growth has been widely studied. However, the details on the type of heterocellular cross-talk between stromal and breast cancer cells (BCCs) are still poorly known. In the present study, in order to investigate the intercellular communication between human mesenchymal stromal cells (hMSCs) and breast cancer cells (BCCs, MDA-MB-231), we recruited cell-internalizing quantum dots (i-QD) generated by conjugation of cell-internalizing anti-mortalin antibody and quantum dots (QD). Co-culture of illuminated and color-coded hMSCs (QD655) and BCCs (QD585) revealed the intercellular transfer of QD655 signal from hMSCs to BCCs. The amount of QD double positive BCCs increased gradually within 48 h of co-culture. We found prominent intercellular transfer of QD655 in hanging drop co-culture system and it was non-existent when hMSCs and BBCs cells were co-cultured in trans-well system lacking imminent cell–cell contact. Fluorescent and electron microscope analyses also supported that the direct cell-to-cell interactions may be required for the intercellular transfer of QD655 from hMSCs to BCCs. To the best of our knowledge, the study provides a first demonstration of transcellular crosstalk between stromal cells and BCCs that involve direct contact and may also include a transfer of mortalin, an anti-apoptotic and growth-promoting factor enriched in cancer cells.

  16. Deoxyelephantopin from Elephantopus scaber L. induces cell-cycle arrest and apoptosis in the human nasopharyngeal cancer CNE cells

    SciTech Connect (OSTI)

    Su, Miaoxian [Biology Programme (Formally Biology Dept.), School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR (China)] [Biology Programme (Formally Biology Dept.), School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR (China); Chung, Hau Yin, E-mail: anthonychung@cuhk.edu.hk [Biology Programme (Formally Biology Dept.), School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR (China); Food and Nutritional Sciences Programme, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong SAR (China); Li, Yaolan [Institute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan University, Guangzhou (China) [Institute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan University, Guangzhou (China); Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drug Research, Guangzhou (China)

    2011-07-29

    Highlights: {yields} Deoxyelephantopin (ESD) inhibited cell proliferation in the human nasopharyngeal cancer CNE cells. {yields} ESD induced cell cycle arrest in S and G2/M phases via modulation of cell cycle regulatory proteins. {yields} ESD triggered apoptosis by dysfunction of mitochondria and induction of both intrinsic and extrinsic apoptotic signaling pathways. {yields} ESD also triggered Akt, ERK, and JNK signaling pathways. -- Abstract: Deoxyelephantopin (ESD), a naturally occurring sesquiterpene lactone present in the Chinese medicinal herb, Elephantopus scaber L. exerted anticancer effects on various cultured cancer cells. However, the cellular mechanisms by which it controls the development of the cancer cells are unavailable, particularly the human nasopharyngeal cancer CNE cells. In this study, we found that ESD inhibited the CNE cell proliferation. Cell cycle arrest in S and G2/M phases was also found. Western blotting analysis showed that modulation of cell cycle regulatory proteins was responsible for the ESD-induced cell cycle arrest. Besides, ESD also triggered apoptosis in CNE cells. Dysfunction in mitochondria was found to be associated with the ESD-induced apoptosis as evidenced by the loss of mitochondrial membrane potential ({Delta}{Psi}m), the translocation of cytochrome c, and the regulation of Bcl-2 family proteins. Despite the Western blotting analysis showed that both intrinsic and extrinsic apoptotic pathways (cleavage of caspases-3, -7, -8, -9, and -10) were triggered in the ESD-induced apoptosis, additional analysis also showed that the induction of apoptosis could be achieved by the caspase-independent manner. Besides, Akt, ERK and JNK pathways were found to involve in ESD-induced cell death. Overall, our findings provided the first evidence that ESD induced cell cycle arrest, and apoptosis in CNE cells. ESD could be a potential chemotherapeutic agent in the treatment of nasopharyngeal cancer (NPC).

  17. Accelerated cellular senescence phenotype of GAPDH-depleted human lung carcinoma cells

    SciTech Connect (OSTI)

    Phadke, Manali; Krynetskaia, Natalia; Mishra, Anurag; Krynetskiy, Evgeny; Jayne Haines Center for Pharmacogenomics, Temple University School of Pharmacy, Philadelphia, PA 19140

    2011-07-29

    Highlights: {yields} We examined the effect of glyceraldehyde 3-phosphate (GAPDH) depletion on proliferation of human carcinoma A549 cells. {yields} GAPDH depletion induces accelerated senescence in tumor cells via AMPK network, in the absence of DNA damage. {yields} Metabolic and genetic rescue experiments indicate that GAPDH has regulatory functions linking energy metabolism and cell cycle. {yields} Induction of senescence in LKB1-deficient lung cancer cells via GAPDH depletion suggests a novel strategy to control tumor cell proliferation. -- Abstract: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a pivotal glycolytic enzyme, and a signaling molecule which acts at the interface between stress factors and the cellular apoptotic machinery. Earlier, we found that knockdown of GAPDH in human carcinoma cell lines resulted in cell proliferation arrest and chemoresistance to S phase-specific cytotoxic agents. To elucidate the mechanism by which GAPDH depletion arrests cell proliferation, we examined the effect of GAPDH knockdown on human carcinoma cells A549. Our results show that GAPDH-depleted cells establish senescence phenotype, as revealed by proliferation arrest, changes in morphology, SA-{beta}-galactosidase staining, and more than 2-fold up-regulation of senescence-associated genes DEC1 and GLB1. Accelerated senescence following GAPDH depletion results from compromised glycolysis and energy crisis leading to the sustained AMPK activation via phosphorylation of {alpha} subunit at Thr172. Our findings demonstrate that GAPDH depletion switches human tumor cells to senescent phenotype via AMPK network, in the absence of DNA damage. Rescue experiments using metabolic and genetic models confirmed that GAPDH has important regulatory functions linking the energy metabolism and the cell cycle networks. Induction of senescence in LKB1-deficient non-small cell lung cancer cells via GAPDH depletion suggests a novel strategy to control tumor cell proliferation.

  18. Osmotic stress affects functional properties of human melanoma cell lines

    E-Print Network [OSTI]

    La Porta, Caterina A M; Pasini, Maria; Laurson, Lasse; Alava, Mikko J; Zapperi, Stefano; Amar, Martine Ben

    2015-01-01

    Understanding the role of microenvironment in cancer growth and metastasis is a key issue for cancer research. Here, we study the effect of osmotic pressure on the functional properties of primary and metastatic melanoma cell lines. In particular, we experimentally quantify individual cell motility and transmigration capability. We then perform a circular scratch assay to study how a cancer cell front invades an empty space. Our results show that primary melanoma cells are sensitive to a low osmotic pressure, while metastatic cells are less. To better understand the experimental results, we introduce and study a continuous model for the dynamics of a cell layer and a stochastic discrete model for cell proliferation and diffusion. The two models capture essential features of the experimental results and allow to make predictions for a wide range of experimentally measurable parameters.

  19. Reprogramming of Human Peripheral Blood Cells to Induced Pluripotent Stem Cells

    E-Print Network [OSTI]

    Staerk, Judith

    Embryonic stem cells are pluripotent cells derived from the inner cell mass of the developing embryo that have the capacity to differentiate into every cell type of the adult (Evans and Kaufman, 1981, Martin, 1981, Martin ...

  20. Detection of human hematopoietic stem cell engraftment in the livers of adult immunodeficient mice by an optimized flow cytometric method

    E-Print Network [OSTI]

    Varga, Nicole L.; Bárcena, Alicia; Fomin, Marina E.; Muench, Marcus O.

    2010-01-01

    CD19 + CD34 + cells, indicating that the full range of humanfull Figure 9. Detection of human B-lymphoid cells in mousefull lymph- oid reconstitution 229 days after FBM trans- plantation are shown in Figure 10. T-cells

  1. How Plasmodium falciparum malaria parasites bind to human brain endothelial cells 

    E-Print Network [OSTI]

    Claessens, Antoine

    2011-01-01

    Cerebral malaria is characterised by an accumulation of infected erythrocytes in the microvasculature of the brain. Plasmodium falciparum infected erythrocytes have been shown to bind to a Human Brain Endothelial Cell line (HBEC-5i) in vitro...

  2. Direct and indirect effects of alpha-particle irradiations of human prostate tumor cells

    E-Print Network [OSTI]

    Wang, Rong, Ph. D. Massachusetts Institute of Technology

    2005-01-01

    The objective of this project is to establish a model system to study the direct effect, the bystander effect and the combinational effect of alpha-particle irradiations of human prostate tumor cells, toward the goal of ...

  3. Role for oestrogen in dynamic interactions between cell types within the human endometrium 

    E-Print Network [OSTI]

    Gibson, Douglas Alistair

    2012-11-30

    The human endometrium is a complex multicellular tissue, located within the cavity of the uterus. Its luminal surface is defined by a layer of epithelial cells supported on a multicellular stroma containing fibroblasts, ...

  4. Soluble interleukin 2 receptors are released from activated human lymphoid cells in vitro

    SciTech Connect (OSTI)

    Rubin, L.A.; Kurman, C.C.; Fritz, M.E.; Biddison, W.E.; Boutin, B.; Yarchoan, R.; Nelson, D.L.

    1985-11-01

    With the use of an enzyme-linked immunoabsorbent assay to measure soluble human interleukin 2 receptors (IL 2R), certain human T cell leukemia virus I (HTLV I)-positive T cell lines were found to spontaneously release large quantities of IL 2R into culture supernatants. This was not found with HTLV I-negative and IL 2 independent T cell lines, and only one of seven B cell-derived lines examined produced small amounts of IL 2R. In addition to this constitutive production of soluble IL 2R by certain cell lines, normal human peripheral blood mononuclear cells (PBMC) could be induced to release soluble IL 2R by plant lectins, the murine monoclonal antibody OKT3, tetanus toxoid, and allogeneic cells. Such activated cells also expressed cellular IL 2R measurable in detergent solubilized cell extracts. The generation of cellular and supernatant IL 2R was: dependent on cellular activation, rapid, radioresistant (3000 rad), and inhibited by cycloheximide treatment. NaDodSO4-polyacrylamide gel electrophoresis analysis of soluble IL 2R demonstrated molecules of apparent Mr = 35,000 to 40,000, and 45,000 to 50,000, respectively, somewhat smaller than the mature surface receptor on these cells. The release of soluble IL 2R appears to be a characteristic marker of T lymphocyte activation and might serve an immunoregulatory function during both normal and abnormal cell growth and differentiation.

  5. 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.

  6. Human natural killer cells control Plasmodium falciparum infection by eliminating infected red blood cells

    E-Print Network [OSTI]

    Chen, Qingfeng

    Immunodeficient mouse–human chimeras provide a powerful approach to study host-specific pathogens, such as Plasmodium falciparum that causes human malaria. Supplementation of immunodeficient mice with human RBCs supports ...

  7. Transcriptome analysis of the human T lymphocyte cell line Jurkat and human peripheral blood mononuclear cells exposed to deoxynivalenol (DON): New mechanistic insights

    SciTech Connect (OSTI)

    Katika, Madhumohan R.; Department of Health Risk Analysis and Toxicology, Maastricht University; Netherlands Toxicogenomics Centre ; Hendriksen, Peter J.M.; Netherlands Toxicogenomics Centre ; Shao, Jia; Department of Health Risk Analysis and Toxicology, Maastricht University; Netherlands Toxicogenomics Centre ; Loveren, Henk van; National Institute for Public Health and the Environment , Bilthoven; Netherlands Toxicogenomics Centre ; Peijnenburg, Ad; Netherlands Toxicogenomics Centre

    2012-10-01

    Deoxynivalenol (DON) or vomitoxin is a commonly encountered type-B trichothecene mycotoxin, produced by Fusarium species predominantly found in cereals and grains. DON is known to exert toxic effects on the gastrointestinal, reproductive and neuroendocrine systems, and particularly on the immune system. Depending on dose and exposure time, it can either stimulate or suppress immune function. The main objective of this study was to obtain a deeper insight into DON-induced effects on lymphoid cells. For this, we exposed the human T-lymphocyte cell line Jurkat and human peripheral blood mononuclear cells (PBMCs) to various concentrations of DON for various times and examined gene expression changes by DNA microarray analysis. Jurkat cells were exposed to 0.25 and 0.5 ?M DON for 3, 6 and 24 h. Biological interpretation of the microarray data indicated that DON affects various processes in these cells: It upregulates genes involved in ribosome structure and function, RNA/protein synthesis and processing, endoplasmic reticulum (ER) stress, calcium-mediated signaling, mitochondrial function, oxidative stress, the NFAT and NF-?B/TNF-? pathways, T cell activation and apoptosis. The effects of DON on the expression of genes involved in ER stress, NFAT activation and apoptosis were confirmed by qRT-PCR. Other biochemical experiments confirmed that DON activates calcium-dependent proteins such as calcineurin and M-calpain that are known to be involved in T cell activation and apoptosis. Induction of T cell activation was also confirmed by demonstrating that DON activates NFATC1 and induces its translocation from the cytoplasm to the nucleus. For the gene expression profiling of PBMCs, cells were exposed to 2 and 4 ?M DON for 6 and 24 h. Comparison of the Jurkat microarray data with those obtained with PBMCs showed that most of the processes affected by DON in the Jurkat cell line were also affected in the PBMCs. -- Highlights: ? The human T cell line Jurkat and human PBMCs were exposed to DON. ? Whole-genome microarray experiments were performed. ? Microarray data indicates that DON affects ribosome and RNA/protein synthesis. ? DON treatment induces ER stress, calcium mediated signaling, NFAT and NF-?B. ? Exposure to DON induces T cell activation, oxidative stress and apoptosis.

  8. KILLING OF TARGET CELLS DUE TO RADON PROGENY IN THE HUMAN LUNG

    E-Print Network [OSTI]

    Yu, Peter K.N.

    KILLING OF TARGET CELLS DUE TO RADON PROGENY IN THE HUMAN LUNG B. M. F. Lau1 , D. Nikezic1,2 and K to inhaled radon progeny in the human lung. The present work uses the microdosimetric approach and determines/alleviate this discrepancy, including those based on different lung morpho- metry models(4) , different ethnic groups(5

  9. Enhancement of P53-Mutant Human Colorectal Cancer Cells Radiosensitivity by Flavonoid Fisetin

    SciTech Connect (OSTI)

    Chen Wenshu [Department of Life Science, Tzu Chi University, Hualien (China); Lee Yijang [Department of Biomedical Imaging and Radiological Sciences, National Yang-Ming University, Taipei (China); Yu Yichu; Hsaio Chinghui [Department of Life Science, Tzu Chi University, Hualien (China)

    2010-08-01

    Purpose: The aim of this study was to investigate whether fisetin is a potential radiosensitizer for human colorectal cancer cells, which are relatively resistant to radiotherapy. Methods and Materials: Cell survival was examined by clonogenic survival assay, and DNA fragmentation was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. The effects of treatments on cell cycle distribution and apoptosis were examined by flow cytometry. Western blot analysis was performed to ascertain the protein levels of {gamma}-H2AX, phospho-Chk2, active caspase-3, PARP cleavage, phospho-p38, phospho-AKT, and phospho-ERK1/2. Results: Fisetin pretreatment enhanced the radiosensitivity of p53-mutant HT-29 human colorectal cancer cells but not human keratocyte HaCaT cells; it also prolonged radiation-induced G{sub 2}/M arrest, enhanced radiation-induced cell growth arrest in HT-29 cells, and suppressed radiation-induced phospho-H2AX (Ser-139) and phospho-Chk2 (Thr-68) in p53-mutant HT-29 cells. Pretreatment with fisetin enhanced radiation-induced caspase-dependent apoptosis in HT-29 cells. Fisetin pretreatment augmented radiation-induced phosphorylation of p38 mitogen-activated protein kinase, which is involved in caspase-mediated apoptosis, and SB202190 significantly reduced apoptosis and radiosensitivity in fisetin-pretreated HT-29 cells. By contrast, both phospho-AKT and phospho-ERK1/2, which are involved in cell proliferation and antiapoptotic pathways, were suppressed after irradiation combined with fisetin pretreatment. Conclusions: To our knowledge, this study is the first to provide evidence that fisetin exerts a radiosensitizing effect in p53-mutant HT-29 cells. Fisetin could potentially be developed as a novel radiosensitizer against radioresistant human cancer cells.

  10. Generation of knock-in primary human T cells using Cas9 ribonucleoproteins

    SciTech Connect (OSTI)

    Schumann, Kathrin; Lin, Steven; Boyer, Eric; Simeonov, Dimitre R.; Subramaniam, Meena; Gate, Rachel E.; Haliburton, Genevieve E.; Ye, Chun J.; Bluestone, Jeffrey A.; Doudna, Jennifer A.; Marson, Alexander

    2015-07-27

    T-cell genome engineering holds great promise for cell-based therapies for cancer, HIV, primary immune deficiencies, and autoimmune diseases, but genetic manipulation of human T cells has been challenging. Improved tools are needed to efficiently “knock out” genes and “knock in” targeted genome modifications to modulate T-cell function and correct disease-associated mutations. CRISPR/Cas9 technology is facilitating genome engineering in many cell types, but in human T cells its efficiency has been limited and it has not yet proven useful for targeted nucleotide replacements. Here we report efficient genome engineering in human CD4+ T cells using Cas9:single-guide RNA ribonucleoproteins (Cas9 RNPs). Cas9 RNPs allowed ablation of CXCR4, a coreceptor for HIV entry. Cas9 RNP electroporation caused up to ~40% of cells to lose high-level cell-surface expression of CXCR4, and edited cells could be enriched by sorting based on low CXCR4 expression. Importantly, Cas9 RNPs paired with homology-directed repair template oligonucleotides generated a high frequency of targeted genome modifications in primary T cells. Targeted nucleotide replacement was achieved in CXCR4 and PD-1 (PDCD1), a regulator of T-cell exhaustion that is a validated target for tumor immunotherapy. Deep sequencing of a target site confirmed that Cas9 RNPs generated knock-in genome modifications with up to ~20% efficiency, which accounted for up to approximately one-third of total editing events. These results establish Cas9 RNP technology for diverse experimental and therapeutic genome engineering applications in primary human T cells.

  11. Generation of knock-in primary human T cells using Cas9 ribonucleoproteins

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

    Schumann, Kathrin; Lin, Steven; Boyer, Eric; Simeonov, Dimitre R.; Subramaniam, Meena; Gate, Rachel E.; Haliburton, Genevieve E.; Ye, Chun J.; Bluestone, Jeffrey A.; Doudna, Jennifer A.; et al

    2015-07-27

    T-cell genome engineering holds great promise for cell-based therapies for cancer, HIV, primary immune deficiencies, and autoimmune diseases, but genetic manipulation of human T cells has been challenging. Improved tools are needed to efficiently “knock out” genes and “knock in” targeted genome modifications to modulate T-cell function and correct disease-associated mutations. CRISPR/Cas9 technology is facilitating genome engineering in many cell types, but in human T cells its efficiency has been limited and it has not yet proven useful for targeted nucleotide replacements. Here we report efficient genome engineering in human CD4+ T cells using Cas9:single-guide RNA ribonucleoproteins (Cas9 RNPs). Cas9more »RNPs allowed ablation of CXCR4, a coreceptor for HIV entry. Cas9 RNP electroporation caused up to ~40% of cells to lose high-level cell-surface expression of CXCR4, and edited cells could be enriched by sorting based on low CXCR4 expression. Importantly, Cas9 RNPs paired with homology-directed repair template oligonucleotides generated a high frequency of targeted genome modifications in primary T cells. Targeted nucleotide replacement was achieved in CXCR4 and PD-1 (PDCD1), a regulator of T-cell exhaustion that is a validated target for tumor immunotherapy. Deep sequencing of a target site confirmed that Cas9 RNPs generated knock-in genome modifications with up to ~20% efficiency, which accounted for up to approximately one-third of total editing events. These results establish Cas9 RNP technology for diverse experimental and therapeutic genome engineering applications in primary human T cells.« less

  12. Human serum activates CIDEB-mediated lipid droplet enlargement in hepatoma cells

    SciTech Connect (OSTI)

    Singaravelu, Ragunath; National Research Council of Canada, Ottawa, Ontario K1A 0R6 ; Lyn, Rodney K.; National Research Council of Canada, Ottawa, Ontario K1A 0R6 ; Srinivasan, Prashanth; Delcorde, Julie; National Research Council of Canada, Ottawa, Ontario K1A 0R6 ; Steenbergen, Rineke H.; Tyrrell, D. Lorne; Li Ka Shing Institute of Virology, Katz Centre for Pharmacy and Health Research, Edmonton, Alberta T6G 2S2 ; Pezacki, John P.

    2013-11-15

    Highlights: •Human serum induced differentiation of hepatoma cells increases cellular lipid droplet (LD) size. •The observed increase in LD size correlates with increased PGC-1? and CIDEB expression. •Induction of CIDEB expression correlates with rescue of VLDL secretion and loss of ADRP. •siRNA knockdown of CIDEB impairs the human serum mediated increase in LD size. •This system represents a cost-efficient model to study CIDEB’s role in lipid biology. -- Abstract: Human hepatocytes constitutively express the lipid droplet (LD) associated protein cell death-inducing DFFA-like effector B (CIDEB). CIDEB mediates LD fusion, as well as very-low-density lipoprotein (VLDL) maturation. However, there are limited cell culture models readily available to study CIDEB’s role in these biological processes, as hepatoma cell lines express negligible levels of CIDEB. Recent work has highlighted the ability of human serum to differentiate hepatoma cells. Herein, we demonstrate that culturing Huh7.5 cells in media supplemented with human serum activates CIDEB expression. This activation occurs through the induced expression of PGC-1?, a positive transcriptional regulator of CIDEB. Coherent anti-Stokes Raman scattering (CARS) microscopy revealed a correlation between CIDEB levels and LD size in human serum treated Huh7.5 cells. Human serum treatment also resulted in a rapid decrease in the levels of adipose differentiation-related protein (ADRP). Furthermore, individual overexpression of CIDEB was sufficient to down-regulate ADRP protein levels. siRNA knockdown of CIDEB revealed that the human serum mediated increase in LD size was CIDEB-dependent. Overall, our work highlights CIDEB’s role in LD fusion, and presents a new model system to study the PGC-1?/CIDEB pathway’s role in LD dynamics and the VLDL pathway.

  13. Distribution of Immune Cells in the Human Cervix and Implications for HIV Transmission

    E-Print Network [OSTI]

    Lieberman, Judy

    Distribution of Immune Cells in the Human Cervix and Implications for HIV Transmission Radiana T transmission. Am J Reprod Immunol 2014; 71: 252­264 doi:10.1111/aji.12198 Problem Knowledge of the mucosal a third were CD11c+ , most of which were CD103À CD11b+ CX3CR1+ DC-SIGN+ dendritic cells (DCs). The other

  14. The promoters of human cell cycle genes integrate signals from two tumor suppressive pathways during

    E-Print Network [OSTI]

    Domany, Eytan

    The promoters of human cell cycle genes integrate signals from two tumor suppressive pathways.9.05 Deciphering regulatory events that drive malignant transformation represents a major challenge for systemsINK4A tumor suppressors inactivation. This cluster predominantly consists of cell cycle genes

  15. ERK-dependent and -independent pathways trigger human neural progenitor cell migration

    SciTech Connect (OSTI)

    Moors, Michaela [Institut fuer Umweltmedizinische Forschung gGmbH at the Heinrich Heine-University, Group of Toxicology, Auf'm Hennekamp 50, 40225 Duesseldorf (Germany)]. E-mail: moors@uni-duesseldorf.de; Cline, Jason E. [Institut fuer Umweltmedizinische Forschung gGmbH at the Heinrich Heine-University, Group of Toxicology, Auf'm Hennekamp 50, 40225 Duesseldorf (Germany)]. E-mail: jason.cline@uni-duesseldorf.de; Abel, Josef [Institut fuer Umweltmedizinische Forschung gGmbH at the Heinrich Heine-University, Group of Toxicology, Auf'm Hennekamp 50, 40225 Duesseldorf (Germany)]. E-mail: josef.abel@uni-duesseldorf.de; Fritsche, Ellen [Institut fuer Umweltmedizinische Forschung gGmbH at the Heinrich Heine-University, Group of Toxicology, Auf'm Hennekamp 50, 40225 Duesseldorf (Germany)]. E-mail: ellen.fritsche@uni-duesseldorf.de

    2007-05-15

    Besides differentiation and apoptosis, cell migration is a basic process in brain development in which neural cells migrate several centimeters within the developing brain before reaching their proper positions and forming the right connections. For identifying signaling events that control neural migration and are therefore potential targets of chemicals to disturb normal brain development, we developed a human neurosphere-based migration assay based on normal human neural progenitor (NHNP) cells, in which the distance is measured that cells wander over time. Applying this assay, we investigated the role of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in the regulation of NHNP cell migration. Exposure to model substances like ethanol or phorbol 12-myristate 13-acetate (PMA) revealed a correlation between ERK1/2 activation and cell migration. The participation of phospho-(P-) ERK1/2 was confirmed by exposure of the cells to the MEK inhibitor PD98059, which directly prohibits ERK1/2 phosphorylation and inhibited cell migration. We identified protein kinase C (PKC) and epidermal growth factor receptor (EGFR) as upstream signaling kinases governing ERK1/2 activation, thereby controlling NHNP cell migration. Additionally, treatments with src kinase inhibitors led to a diminished cell migration without affecting ERK1/2 phosphorylation. Based on these results, we postulate that migration of NHNP cells is controlled via ERK1/2-dependent and -independent pathways.

  16. PIAS? Is Required for Faithful Chromosome Segregation in Human Cells

    E-Print Network [OSTI]

    Dí az-Martí nez, Laura A.; Gimé nez-Abiá n, Juan F.; Azuma, Yoshiaki; Guacci, Vincent; Gimé nez-Martí n, Gonzalo; Lanier, Lorene M.; Clarke, Duncan J.

    2006-12-20

    . In cells lacking PIAS?, normal metaphase plates form, but the spindle checkpoint is activated, leading to a prolonged metaphase block. Sister chromatids remain cohered even if cohesin is removed by depletion of hSgo1, because DNA catenations persist...

  17. Endothelial influences enhance human pluripotent stem cell -derived cardiomyocyte maturation

    E-Print Network [OSTI]

    Wei, Karen A.

    2011-01-01

    hPSC EBs are plated on gelatin- coated tissue culturewere plated on 0.1% gelatin-coated (Stem Cell Technologies,EBs were plated on 0.1% gelatin-coated tissue culture dishes

  18. Peripheral blood derived mononuclear cells enhance osteoarthritic human chondrocyte migration

    E-Print Network [OSTI]

    Hopper, Niina; Henson, Frances; Brooks, Roger; Ali, Erden; Rushton, Neil; Wardale, John

    2015-01-01

    peripheral blood mononuclear cells stimulated by LPS or infected by Borrelia. Physiological research / Academia Scientiarum Bohemoslovaca. 2003;52(6):593-8. 23. Mildner M, Hacker S, Haider T, Gschwandtner M, Werba G, Barresi C et al. Secretome...

  19. Activation of ERK mitogen-activated protein kinase in human cells by the mycotoxin patulin

    SciTech Connect (OSTI)

    Wu, T.-S. [Department of Life Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Road, Taichung, Taiwan (China); Yu, F.-Y. [Department of Life Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Road, Taichung, Taiwan (China); Su, C.-C. [Tian-Sheng Memorial Hospital, Tong kong, Ping-Tong, Taiwan (China); Kan, J.-C. [Department of Life Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Road, Taichung, Taiwan (China); Chung, C.-P. [Department of Life Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Road, Taichung, Taiwan (China); Liu, B.-H. [Department of Life Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Road, Taichung, Taiwan (China)]. E-mail: bingliu@csmu.edu.tw

    2005-09-01

    Patulin (PAT), a mycotoxin produced by certain species of Penicillium and Aspergillus, is often detectable in moldy fruits and their derivative products. PAT led to a concentration-dependent and time-dependent increase in phosphorylation of extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) in human embryonic kidney (HEK293) cells, human peripheral blood mononuclear cells (PBMCs), and Madin-Darby canine kidney (MDCK) cells. Exposure of HEK293 cells to concentrations above 5 {mu}M PAT for 30 min induced ERK1/2 phosphorylation; activation of ERK1/2 was also observed after 24 h incubation with 0.05 {mu}M of PAT. Treatment of human PBMCs for 30 min with 30 {mu}M PAT dramatically increased the phosphorylated ERK1/2 levels. Both MEK1/2 inhibitors, U0126 and PD98059, suppressed ERK1/2 activation in either HEK293 or MDCK cells. In HEK293 cells, U0126-mediated inhibition of PAT-induced ERK1/2 phosphorylation resulted in a significant decrease in levels of DNA damage, expressed as tail moment values, in the single cell gel electrophoresis assay. Conversely, U0126 did not affect cell viability, lactate dehydrogenase release, and the DNA synthesis rate in PAT-treated cultures. Exposure of HEK293 cells for 90 min to 15 {mu}M PAT elevated the levels of early growth response gene-1 (egr-1) mRNA, but not of c-fos, fosB, and junB mRNAs. These results indicate that in human cells, PAT causes a rapid and persistent activation of ERK1/2 and this signaling pathway plays an important role in mediating PAT-induced DNA damage and egr-1 gene expression.

  20. Enhanced growth medium and method for culturing human mammary epithelial cells

    DOE Patents [OSTI]

    Stampfer, Martha R. (7290 Sayre Dr., Oakland, CA 94611); Smith, Helene S. (5693 Cabot Dr., Oakland, CA 94611); Hackett, Adeline J. (82 Evergreen Dr., Orinda, CA 94563)

    1983-01-01

    Methods are disclosed for isolating and culturing human mammary epithelial cells of both normal and malignant origin. Tissue samples are digested with a mixture including the enzymes collagenase and hyaluronidase to produce clumps of cells substantially free from stroma and other undesired cellular material. Growing the clumps of cells in mass culture in an enriched medium containing particular growth factors allows for active cell proliferation and subculture. Clonal culture having plating efficiencies of up to 40% or greater may be obtained using individual cells derived from the mass culture by plating the cells on appropriate substrates in the enriched media. The clonal growth of cells so obtained is suitable for a quantitative assessment of the cytotoxicity of particular treatment. An exemplary assay for assessing the cytotoxicity of the drug adriamycin is presented.

  1. Development and Investigation of Synthetic Skin Simulant Platform (3SP) in Friction Blister Applications 

    E-Print Network [OSTI]

    Guerra, Carlos

    2012-02-14

    Skin is the largest organ of the human body. It is the first line of defense between the vulnerable organs and tissues of the body and the environment. Healthy skin is paramount to avoiding infection and disease. Therefore, ...

  2. Regulation of E2F-1 gene expression in human breast cancer cells 

    E-Print Network [OSTI]

    Ngwenya, Sharon Khethiwe

    2005-08-29

    Breast Cancer Cells. (May 2005) Sharon Khethiwe Ngwenya, B.S., Oakwood College Chair of Advisory Committee: Dr. Stephen Safe 17beta-Estradiol induces E2F-1 gene expression in ZR-75 and MCF-7 human breast cancer cells. Analysis of the E2F-1 gene... region. This promoter region was also E2-responsive in ERalpha-positive ZR-75 cells; however, further analysis of the promoter showed that cooperative ERalpha/Sp1/NFY interactions were not necessary for hormone-induced transactivation in ZR-75 cells...

  3. Gap Junctions and Connexon Hemichannels in Human Embryonic Stem Cells

    E-Print Network [OSTI]

    Huettner, James E.

    cells main- tained in vitro expressed RNA for 18 of the 20 known connexins; only connexin 40.1 (Cx40.1) and Cx50 were not detected by reverse transcription-polymerase chain reac- tion. Cx40, Cx43, and Cx45 communication that is observed in early embryos [3, 4]. More than 20 different connexin (Cx) subunits

  4. Mechanosensitivity of human osteosarcoma cells and phospholipase C {beta}2 expression

    SciTech Connect (OSTI)

    Hoberg, M. [Department of Orthopaedics, University of Tuebingen (Germany)]. E-mail: Maik.Hoberg@med.uni-tuebingen.de; Gratz, H.-H. [Experimental Orthopaedics and Biomechanics, Phillips-University of Marburg (Germany); Noll, M. [Experimental Orthopaedics and Biomechanics, Phillips-University of Marburg (Germany); Jones, D.B. [Experimental Orthopaedics and Biomechanics, Phillips-University of Marburg (Germany)

    2005-07-22

    Bone adapts to mechanical load by osteosynthesis, suggesting that osteoblasts might respond to mechanical stimuli. We therefore investigated cell proliferation and phospholipase C (PLC) expression in osteoblasts. One Hertz uniaxial stretching at 4000 {mu}strains significantly increased the proliferation rates of human osteoblast-like osteosarcoma cell line MG-63 and primary human osteoblasts. However, U-2/OS, SaOS-2, OST, and MNNG/HOS cells showed no significant changes in proliferation rate. We investigated the expression pattern of different isoforms of PLC in these cell lines. We were able to detect PLC {beta}1, {beta}3, {gamma}1, {gamma}2, and {delta}1 in all cells, but PLC {beta}2 was only detectable in the mechanosensitive cells. We therefore investigated the possible role of PLC {beta}2 in mechanotransduction. Inducible antisense expression for 24 h inhibited the translation of PLC {beta}1 in U-2/OS cells by 35% and PLC {beta}2 in MG-63 by 29%. Fluid shear flow experiments with MG-63 lacking PLC {beta}2 revealed a significantly higher level of cells losing attachment to coverslips and a significantly lower number of cells increasing intracellular free calcium.

  5. Distinct p53 genomic binding patterns in normal and cancer-derived human cells

    SciTech Connect (OSTI)

    Botcheva K.; McCorkle S. R.; McCombie W. R.; Dunn J. J.; Anderson C. W.

    2011-12-15

    We report here genome-wide analysis of the tumor suppressor p53 binding sites in normal human cells. 743 high-confidence ChIP-seq peaks representing putative genomic binding sites were identified in normal IMR90 fibroblasts using a reference chromatin sample. More than 40% were located within 2 kb of a transcription start site (TSS), a distribution similar to that documented for individually studied, functional p53 binding sites and, to date, not observed by previous p53 genome-wide studies. Nearly half of the high-confidence binding sites in the IMR90 cells reside in CpG islands, in marked contrast to sites reported in cancer-derived cells. The distinct genomic features of the IMR90 binding sites do not reflect a distinct preference for specific sequences, since the de novo developed p53 motif based on our study is similar to those reported by genome-wide studies of cancer cells. More likely, the different chromatin landscape in normal, compared with cancer-derived cells, influences p53 binding via modulating availability of the sites. We compared the IMR90 ChIPseq peaks to the recently published IMR90 methylome1 and demonstrated that they are enriched at hypomethylated DNA. Our study represents the first genome-wide, de novo mapping of p53 binding sites in normal human cells and reveals that p53 binding sites reside in distinct genomic landscapes in normal and cancer-derived human cells.

  6. Mechanisms of Telomere Protection and Deprotection in Human Cells

    E-Print Network [OSTI]

    Sarthy, Jay Francis

    2009-07-31

    ………..77 e. Telomeric hRAP1 Counteracts Uncapping by TRF2????…....79 4. Discussion……………………………………………………………….85 9 Chapter IV. Replication Stress at Telomeres Results in “t-wrecks”, a New Type of Highly Conserved Telomere Dysfunction…………………………………….91 1... are considered to be the fundamental unit of life on Earth. The first description of cells is from the 17 th century polymath Robert Hooke. While inspecting cork bark at high magnification using the newly invented microscope, Hooke famously noted...

  7. Reconstitution activity of hypoxic cultured human cord blood CD34-positive cells in NOG mice

    SciTech Connect (OSTI)

    Shima, Haruko; Takubo, Keiyo; Iwasaki, Hiroko; Yoshihara, Hiroki; Gomei, Yumiko; Hosokawa, Kentaro; Arai, Fumio; Takahashi, Takao; Suda, Toshio

    2009-01-16

    Hematopoietic stem cells (HSCs) reside in hypoxic areas of the bone marrow. However, the role of hypoxia in the maintenance of HSCs has not been fully characterized. We performed xenotransplantation of human cord blood cells cultured in hypoxic or normoxic conditions into adult NOD/SCID/IL-2R{gamma}{sup null} (NOG) mice. Hypoxic culture (1% O{sub 2}) for 6 days efficiently supported the maintenance of HSCs, although cell proliferation was suppressed compared to the normoxic culture. In contrast, hypoxia did not affect in vitro colony-forming ability. Upregulation of a cell cycle inhibitor, p21, was observed in hypoxic culture. Immunohistochemical analysis of recipient bone marrow revealed that engrafted CD34{sup +}CD38{sup -} cord blood HSCs were hypoxic. Taken together, these results demonstrate the significance of hypoxia in the maintenance of quiescent human cord blood HSCs.

  8. Evidence for toxicity differences between inorganic arsenite and thioarsenicals in human bladder cancer cells

    SciTech Connect (OSTI)

    Naranmandura, Hua [Analytical and Environmental Toxicology, Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada T6G 2G3 (Canada); Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675 (Japan); Ogra, Yasumitsu; Iwata, Katsuya [Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675 (Japan); Lee, Jane [Department of Oncology, Cross Cancer Institute, University of Alberta, Edmonton, Alberta, T6G 1Z2 (Canada); Suzuki, Kazuo T. [Graduate School of Pharmaceutical Sciences, Chiba University, Chuo, Chiba 260-8675 (Japan); Weinfeld, Michael [Department of Oncology, Cross Cancer Institute, University of Alberta, Edmonton, Alberta, T6G 1Z2 (Canada); Le, X. Chris [Analytical and Environmental Toxicology, Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, T6G 2G3 (Canada)], E-mail: xc.le@ualberta.ca

    2009-07-15

    Arsenic toxicity is dependent on its chemical species. In humans, the bladder is one of the primary target organs for arsenic-induced carcinogenicity. However, little is known about the mechanisms underlying arsenic-induced carcinogenicity, and what arsenic species are responsible for this carcinogenicity. The present study aimed at comparing the toxic effect of DMMTA{sup V} with that of inorganic arsenite (iAs{sup III}) on cell viability, uptake efficiency and production of reactive oxygen species (ROS) toward human bladder cancer EJ-1 cells. The results were compared with those of a previous study using human epidermoid carcinoma A431 cells. Although iAs{sup III} was known to be toxic to most cells, here we show that iAs{sup III} (LC{sub 50} = 112 {mu}M) was much less cytotoxic than DMMTA{sup V} (LC{sub 50} = 16.7 {mu}M) in human bladder EJ-1 cells. Interestingly, pentavalent sulfur-containing DMMTA{sup V} generated a high level of intracellular ROS in EJ-1 cells. However, this was not observed in the cells exposed to trivalent inorganic iAs{sup III} at their respective LC{sub 50} dose. Furthermore, the presence of N-acetyl-cysteine completely inhibited the cytotoxicity of DMMTA{sup V} but not iAs{sup III}, suggesting that production of ROS was the main cause of cell death from exposure to DMMTA{sup V}, but not iAs{sup III}. Because the cellular uptake of iAs{sup III} is mediated by aquaporin proteins, and because the resistance of cells to arsenite can be influenced by lower arsenic uptake due to lower expression of aquaporin proteins (AQP 3, 7 and 9), the expression of several members of the aquaporin family was also examined. In human bladder EJ-1 cells, mRNA/proteins of AQP3, 7 and 9 were not detected by reverse transcription polymerase chain reaction (RT-PCR)/western blotting. In A431 cells, only mRNA and protein of AQP3 were detected. The large difference in toxicity between the two cell lines could be related to their differences in uptake of arsenic species.

  9. Elevated expression of ERK 2 in human tumor cells chronically treated with PD98059

    SciTech Connect (OSTI)

    Kanda, Shigeru [Department of Molecular Microbiology and Immunology, Division of Endothelial Cell Biology, Nagasaki University Graduate School of Biomedical Science, Nagasaki (Japan)]. E-mail: skanda-jua@umin.net; Kanetake, Hiroshi [Department of Urology, Nagasaki University Graduate School of Biomedical Science, Nagasaki (Japan); Miyata, Yasuyoshi [Department of Urology, Nagasaki University Graduate School of Biomedical Science, Nagasaki (Japan)

    2006-07-14

    We examined the effect of chronic exposure of tumor cells to a mitogen-activated protein kinase/extracellular signal-regulated kinases (ERK) kinase inhibitor, PD98059, on cell proliferation was investigated. Human renal carcinoma cells (ACHN) and prostatic carcinoma cells (DU145) were cultured in the presence of PD98059 for more than 4 weeks (denoted ACHN (PD) cells and DU145 (PD) cells, respectively) and proliferation and signal transduction pathways were examined. PD98059 significantly inhibited the proliferation of parental cells. However, PD98059 failed to inhibit proliferation of ACHN (PD) and DU145 (PD) cells significantly. Expression of ERK 1 and 2 was elevated in these cells. These phenotypes were reversible. Downregulation of ERK 2, but not ERK 1, by small interfering RNA significantly inhibited the proliferation of ACHN (PD) and DU145 (PD) cells. Taken together, chronic exposure of tumor cells to PD98059 induced elevated expression of ERK 2, which was associated with decreased sensitivity of cellular proliferation to PD98059.

  10. Establishment of three-dimensional cultures of human pancreatic duct epithelial cells

    SciTech Connect (OSTI)

    Gutierrez-Barrera, Angelica M. [Department of Gastrointestinal Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Unit 426, 1515 Holcombe Boulevard, Houston, TX 77030 (United States); Menter, David G. [Department of Thoracic Head and Neck Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, TX (United States); Abbruzzese, James L. [Department of Gastrointestinal Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Unit 426, 1515 Holcombe Boulevard, Houston, TX 77030 (United States); Reddy, Shrikanth A.G. [Department of Gastrointestinal Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Unit 426, 1515 Holcombe Boulevard, Houston, TX 77030 (United States)]. E-mail: sa08366@wotan.mdacc.tmc.edu

    2007-07-06

    Three-dimensional (3D) cultures of epithelial cells offer singular advantages for studies of morphogenesis or the role of cancer genes in oncogenesis. In this study, as part of establishing a 3D culture system of pancreatic duct epithelial cells, we compared human pancreatic duct epithelial cells (HPDE-E6E7) with pancreatic cancer cell lines. Our results show, that in contrast to cancer cells, HPDE-E6E7 organized into spheroids with what appeared to be apical and basal membranes and a luminal space. Immunostaining experiments indicated that protein kinase Akt was phosphorylated (Ser473) and CTMP, a negative Akt regulator, was expressed in both HPDE-E6E7 and cancer cells. However, a nuclear pool of CTMP was detectable in HPDE-E6E7 cells that showed a dynamic concentrated expression pattern, a feature that further distinguished HPDE-E637 cells from cancer cells. Collectively, these data suggest that 3D cultures of HPDE-E6E7 cells are useful for investigating signaling and morphological abnormalities in pancreatic cancer cells.

  11. Generation of human cortical neurons from a new immortal fetal neural stem cell line

    SciTech Connect (OSTI)

    Cacci, E. [Laboratory of Neural Stem Cell Biology, Section of Restorative Neurology, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, BMC B10, Klinikgatan 26, University Hospital, SE-221 84 Lund (Sweden); Villa, A. [Laboratory of Human Neural Stem Cell Research, Center of Molecular Biology Severo Ochoa, Lab CX-450, Autonomous University of Madrid, 28049 Madrid (Spain); Parmar, M. [Division of Neurobiology, Wallenberg Neuroscience Center, Lund University, BMC A11, SE-221 84 Lund (Sweden); Cavallaro, M. [Laboratory of Neural Stem Cell Biology, Section of Restorative Neurology, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, BMC B10, Klinikgatan 26, University Hospital, SE-221 84 Lund (Sweden); Mandahl, N. [Department of Laboratory Medicine, Section of Clinical Genetics, University Hospital, SE-221 85 Lund (Sweden); Lindvall, O. [Laboratory of Neurogenesis and Cell Therapy, Section of Restorative Neurology, Wallenberg Neuroscience Center, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, University Hospital, SE-221 84 Lund (Sweden); Martinez-Serrano, A. [Laboratory of Human Neural Stem Cell Research, Center of Molecular Biology Severo Ochoa, Lab CX-450, Autonomous University of Madrid, 28049 Madrid (Spain); Kokaia, Z. [Laboratory of Neural Stem Cell Biology, Section of Restorative Neurology, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, BMC B10, Klinikgatan 26, University Hospital, SE-221 84 Lund (Sweden)]. E-mail: Zaal.Kokaia@med.lu.se

    2007-02-01

    Isolation and expansion of neural stem cells (NSCs) of human origin are crucial for successful development of cell therapy approaches in neurodegenerative diseases. Different epigenetic and genetic immortalization strategies have been established for long-term maintenance and expansion of these cells in vitro. Here we report the generation of a new, clonal NSC (hc-NSC) line, derived from human fetal cortical tissue, based on v-myc immortalization. Using immunocytochemistry, we show that these cells retain the characteristics of NSCs after more than 50 passages. Under proliferation conditions, when supplemented with epidermal and basic fibroblast growth factors, the hc-NSCs expressed neural stem/progenitor cell markers like nestin, vimentin and Sox2. When growth factors were withdrawn, proliferation and expression of v-myc and telomerase were dramatically reduced, and the hc-NSCs differentiated into glia and neurons (mostly glutamatergic and GABAergic, as well as tyrosine hydroxylase-positive, presumably dopaminergic neurons). RT-PCR analysis showed that the hc-NSCs retained expression of Pax6, Emx2 and Neurogenin2, which are genes associated with regionalization and cell commitment in cortical precursors during brain development. Our data indicate that this hc-NSC line could be useful for exploring the potential of human NSCs to replace dead or damaged cortical cells in animal models of acute and chronic neurodegenerative diseases. Taking advantage of its clonality and homogeneity, this cell line will also be a valuable experimental tool to study the regulatory role of intrinsic and extrinsic factors in human NSC biology.

  12. The FBXW7 {beta}-form is suppressed in human glioma cells

    SciTech Connect (OSTI)

    Gu, Zhaodi; Inomata, Kenichi; Ishizawa, Kota; Horii, Akira . E-mail: horii@mail.tains.tohoku.ac.jp

    2007-03-23

    FBXW7 (F-box and WD40 domain protein 7) is an F-box protein with 7 tandem WDs (tryptophan-aspartic acid) that functions as a phosphoepitope-specific substrate recognition component of SCF (Skp1-Cul1-F-box protein) ubiquitin ligases and catalyzes the ubiquitination of proteins promoting cell proliferation, such as CCNE1, MYC, AURKA, NOTCH1, and JUN, which are frequently activated in a wide range of human cancers. FBXW7 is a candidate tumor suppressor, and mutations have been reported in some human tumors. In this study, we analyzed 84 human tumor cell lines in search for genetic alterations of FBXW7, as well as mRNA and protein expressional changes, and compared them with expression levels of the CCNE1, MYC, and AURKA proteins. We found a novel nonsense mutation in a colon cancer cell line SCC and confirmed the missense mutations in SKOV3, an ovarian cancer cell line, and LoVo, a colon cancer cell line. Moreover, suppressed expression of FBXW7 accompanied by activation of the target proteins were observed in ovarian, colon, endometrial, gastric, and prostate cancers. It is notable that highly suppressed mRNA expression of the FBXW7 {beta}-form was found in all the human glioma cell lines analyzed; enhanced expressions of CCNE1, MYC, and AURKA were observed in these cells. Our present results imply that FBXW7 plays a pivotal role in many tissues by controlling the amount of cell cycle promoter proteins and that dysfunction of this protein is one of the essential steps in carcinogenesis in multiple organs.

  13. Triptolide, an Inhibitor of the Human Heat Shock Response That Enhances Stress-induced Cell Death*S

    E-Print Network [OSTI]

    Morimoto, Richard

    Triptolide, an Inhibitor of the Human Heat Shock Response That Enhances Stress-induced Cell Death chaperones, inducible by heat shock and a variety of other stresses, have critical roles in protein homeostasis, balancing cell stress with adaptation, survival, and cell death mechanisms. In transformed cells

  14. HISTORY OF SKIN GRAFTS and Hauben and colleagues2

    E-Print Network [OSTI]

    Stanford University

    report of successful pinch grafts. Ollier in 1872 pointed out the importance of the dermis in skin grafts, Vandeput, and Olley4 gave us the technology to expand skin grafts with a machine that would cut the graft technology was published by Rheinwald and Green,5 and in 1979 cultured human keratinocytes were grown to form

  15. A human breast cell model of pre-invasive to invasive transition

    SciTech Connect (OSTI)

    Bissell, Mina J; Rizki, Aylin; Weaver, Valerie M.; Lee, Sun-Young; Rozenberg, Gabriela I.; Chin, Koei; Myers, Connie A.; Bascom, Jamie L.; Mott, Joni D.; Semeiks, Jeremy R.; Grate, Leslie R.; Mian, I. Saira; Borowsky, Alexander D.; Jensen, Roy A.; Idowu, Michael O.; Chen, Fanqing; Chen, David J.; Petersen, Ole W.; Gray, Joe W.; Bissell, Mina J.

    2008-03-10

    A crucial step in human breast cancer progression is the acquisition of invasiveness. There is a distinct lack of human cell culture models to study the transition from pre-invasive to invasive phenotype as it may occur 'spontaneously' in vivo. To delineate molecular alterations important for this transition, we isolated human breast epithelial cell lines that showed partial loss of tissue polarity in three-dimensional reconstituted-basement membrane cultures. These cells remained non-invasive; however, unlike their non-malignant counterparts, they exhibited a high propensity to acquire invasiveness through basement membrane in culture. The genomic aberrations and gene expression profiles of the cells in this model showed a high degree of similarity to primary breast tumor profiles. The xenograft tumors formed by the cell lines in three different microenvironments in nude mice displayed metaplastic phenotypes, including squamous and basal characteristics, with invasive cells exhibiting features of higher grade tumors. To find functionally significant changes in transition from pre-invasive to invasive phenotype, we performed attribute profile clustering analysis on the list of genes differentially expressed between pre-invasive and invasive cells. We found integral membrane proteins, transcription factors, kinases, transport molecules, and chemokines to be highly represented. In addition, expression of matrix metalloproteinases MMP-9,-13,-15,-17 was up regulated in the invasive cells. Using siRNA based approaches, we found these MMPs to be required for the invasive phenotype. This model provides a new tool for dissection of mechanisms by which pre-invasive breast cells could acquire invasiveness in a metaplastic context.

  16. Single-cell analysis of the dynamics and functional outcomes of interactions between human natural killer cells and target cells

    E-Print Network [OSTI]

    Yamanaka, Yvonne Joy

    Natural killer (NK) cells are a subset of innate immune lymphocytes that interrogate potential target cells and rapidly respond by lysing them or secreting inflammatory immunomodulators. Productive interactions between NK ...

  17. Omalizumab may decrease IgE synthesis by targeting membrane IgE+ human B cells

    E-Print Network [OSTI]

    Chan, Marcia A.; Gigliotti, Nicole M.; Dotson, Abby Louise; Rosenwasser, Lanny J.

    2013-09-02

    Omalizumab, is a humanized anti-IgE monoclonal antibody used to treat allergic asthma. Decreased serum IgE levels, lower eosinophil and B cell counts have been noted as a result of treatment. In vitro studies and animal models support the hypothesis...

  18. Discovering functional transcription-factor combinations in the human cell cycle

    E-Print Network [OSTI]

    Church, George M.

    Discovering functional transcription-factor combinations in the human cell cycle Zhou Zhu,1 Jay-factor (TF) combinations using phylogenetically conserved sequences and microarray-based expression data of binding sites in the vicinity of one another and whether these combinations result in more coherent

  19. IL-8 Production in Human Lung Fibroblasts and Epithelial Cells Activated by the Pseudomonas Autoinducer

    E-Print Network [OSTI]

    Springer, Timothy A.

    IL-8 Production in Human Lung Fibroblasts and Epithelial Cells Activated by the Pseudomonas is caused, in part, by the produc- tion of the chemokine IL-8, which recruits neutrophils into the lung. These findings support the concept that the severe lung damage that accompanies P. aeruginosa infections

  20. Induction of apoptosis by epigallocatechin-3-gallate in human lymphoblastoid B cells

    SciTech Connect (OSTI)

    Noda, Chiseko He, Jinsong; Takano, Tomoko; Tanaka, Chisato; Kondo, Toshinori; Tohyama, Kaoru; Yamamura, Hirohei; Tohyama, Yumi

    2007-11-03

    (-)-Epigallocatechin-3-gallate (EGCG), a major constituent of green tea polyphenols, has been shown to suppress cancer cell proliferation and induce apoptosis. In this study we investigated its efficacy and the mechanism underlying its effect using human B lymphoblastoid cell line Ramos, and effect of co-treatment with EGCG and a chemotherapeutic agent on apoptotic cell death. EGCG induced dose- and time-dependent apoptotic cell death accompanied by loss of mitochondrial transmembrane potential, release of cytochrome c into the cytosol, and cleavage of pro-caspase-9 to its active form. EGCG also enhanced production of intracellular reactive oxygen species (ROS). Pretreatment with diphenylene iodonium chloride, an inhibitor of NAD(P)H oxidase and an antioxidant, partially suppressed both EGCG-induced apoptosis and production of ROS, implying that oxidative stress is involved in the apoptotic response. Furthermore, we showed that combined-treatment with EGCG and a chemotherapeutic agent, etoposide, synergistically induced apoptosis in Ramos cells.

  1. Calcitriol inhibits Ether-a go-go potassium channel expression and cell proliferation in human breast cancer cells

    SciTech Connect (OSTI)

    Garcia-Becerra, Rocio [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)] [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico); Diaz, Lorenza, E-mail: lorenzadiaz@gmail.com [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)] [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico); Camacho, Javier [Department of Pharmacology, Centro de Investigacion y de Estudios Avanzados, Instituto Politecnico Nacional, Av. Instituto Politecnico Nacional 2508, San Pedro Zacatenco 07360, Mexico, D.F. (Mexico)] [Department of Pharmacology, Centro de Investigacion y de Estudios Avanzados, Instituto Politecnico Nacional, Av. Instituto Politecnico Nacional 2508, San Pedro Zacatenco 07360, Mexico, D.F. (Mexico); Barrera, David; Ordaz-Rosado, David; Morales, Angelica [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)] [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico); Ortiz, Cindy Sharon [Department of Pathology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)] [Department of Pathology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico); Avila, Euclides [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)] [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico); Bargallo, Enrique [Department of Breast Tumors, Instituto Nacional de Cancerologia, Av. San Fernando No. 22, Tlalpan 14080, Mexico, D.F. (Mexico)] [Department of Breast Tumors, Instituto Nacional de Cancerologia, Av. San Fernando No. 22, Tlalpan 14080, Mexico, D.F. (Mexico); Arrecillas, Myrna [Department of Pathology, Instituto Nacional de Cancerologia, Av. San Fernando No. 22, Tlalpan 14080, Mexico, D.F. (Mexico)] [Department of Pathology, Instituto Nacional de Cancerologia, Av. San Fernando No. 22, Tlalpan 14080, Mexico, D.F. (Mexico); Halhali, Ali; Larrea, Fernando [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)] [Department of Reproductive Biology, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Vasco de Quiroga No. 15, Tlalpan 14000 Mexico, D.F. (Mexico)

    2010-02-01

    Antiproliferative actions of calcitriol have been shown to occur in many cell types; however, little is known regarding the molecular basis of this process in breast carcinoma. Ether-a-go-go (Eag1) potassium channels promote oncogenesis and are implicated in breast cancer cell proliferation. Since calcitriol displays antineoplastic effects while Eag1 promotes tumorigenesis, and both factors antagonically regulate cell cycle progression, we investigated a possible regulatory effect of calcitriol upon Eag1 as a mean to uncover new molecular events involved in the antiproliferative activity of this hormone in human breast tumor-derived cells. RT real-time PCR and immunocytochemistry showed that calcitriol suppressed Eag1 expression by a vitamin D receptor (VDR)-dependent mechanism. This effect was accompanied by inhibition of cell proliferation, which was potentiated by astemizole, a nonspecific Eag1 inhibitor. Immunohistochemistry and Western blot demonstrated that Eag1 and VDR abundance was higher in invasive-ductal carcinoma than in fibroadenoma, and immunoreactivity of both proteins was located in ductal epithelial cells. Our results provide evidence of a novel mechanism involved in the antiproliferative effects of calcitriol and highlight VDR as a cancer therapeutic target for breast cancer treatment and prevention.

  2. Monoclonal antibodies to human hemoglobin S and cell lines for the production thereof

    DOE Patents [OSTI]

    Jensen, Ronald H. (Livermore, CA); Vanderlaan, Martin (San Ramon, CA); Bigbee, William L. (Livermore, CA); Stanker, Larry H. (Livermore, CA); Branscomb, Elbert W. (Walnut Creek, CA); Grabske, Robert J. (Berkeley, CA)

    1988-01-01

    The present invention provides monoclonal antibodies specific to and distinguish between hemoglobin S and hemoglobin A and methods for their production and use. These antibodies are capable of distinguishing between two hemoglobin types which differ from each other by only a single amino acid residue. The antibodies produced according to the present method are useful as immunofluorescent markers to enumerate circulating red blood cells which have the property of altered expression of the hemoglobin gene due to somatic mutation in stem cells. Such a measurement is contemplated as an assay for in vivo cellular somatic mutations in humans. Since the monoclonal antibodies produced in accordance with the instant invention exhibit a high degree of specificity to and greater affinity for hemoglobin S, they are suitable for labeling human red blood cells for flow cytometric detection of hemoglobin genotype.

  3. Monoclonal antibodies to human hemoglobin S and cell lines for the production thereof

    DOE Patents [OSTI]

    Jensen, R.H.; Vanderlaan, M.; Bigbee, W.L.; Stanker, L.H.; Branscomb, E.W.; Grabske, R.J.

    1984-11-29

    The present invention provides monoclonal antibodies specific to and distinguishing between hemoglobin S and hemoglobin A and methods for their production and use. These antibodies are capable of distinguishing between two hemoglobin types which differ from each other by only a single amino acid residue. The antibodies produced according to the present method are useful as immunofluorescent markers to enumerate circulating red blood cells which have the property of altered expression of the hemoglobin gene due to somatic mutation in stem cells. Such a measurement is contemplated as an assay for in vivo cellular somatic mutations in humans. Since the monoclonal antibodies produced in accordance with the instant invention exhibit a high degree of specificity to and greater affinity for hemoglobin S, they are suitable for labeling human red blood cells for flow cytometric detection of hemoglobin genotype. 4 figs.

  4. STAT3 signaling pathway is necessary for cell survival and tumorsphere forming capacity in ALDH{sup +}/CD133{sup +} stem cell-like human colon cancer cells

    SciTech Connect (OSTI)

    Lin, Li; Division of Cardiology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030 ; Fuchs, James; Li, Chenglong; Olson, Veronica; Bekaii-Saab, Tanios; Lin, Jiayuh

    2011-12-16

    Highlights: Black-Right-Pointing-Pointer The phosphorylated or activated form of STAT3 was expressed in colon cancer stem-like cells. Black-Right-Pointing-Pointer STAT3 inhibitor, FLLL32 inhibits P-STAT3 and STAT3 target genes in colon cancer stem-like cells. Black-Right-Pointing-Pointer Inhibition of STAT3 resulted in decreased cell viability and reduced numbers of tumorspheres. Black-Right-Pointing-Pointer STAT3 is required for survival and tumorsphere forming capacity in colon cancer stem-like cells. Black-Right-Pointing-Pointer Targeting STAT3 in cancer stem-like cells may offer a novel treatment approach for colon cancer. -- Abstract: Persistent activation of Signal Transducers and Activators of Transcription 3 (STAT3) is frequently detected in colon cancer. Increasing evidence suggests the existence of a small population of colon cancer stem or cancer-initiating cells may be responsible for tumor initiation, metastasis, and resistance to chemotherapy and radiation. Whether STAT3 plays a role in colon cancer-initiating cells and the effect of STAT3 inhibition is still unknown. Flow cytometry was used to isolate colon cancer stem-like cells from three independent human colon cancer cell lines characterized by both aldehyde dehydrogenase (ALDH)-positive and CD133-positive subpopulation (ALDH{sup +}/CD133{sup +}). The effects of STAT3 inhibition in colon cancer stem-like cells were examined. The phosphorylated or activated form of STAT3 was expressed in colon cancer stem-like cells and was reduced by a STAT3-selective small molecular inhibitor, FLLL32. FLLL32 also inhibited the expression of potential STAT3 downstream target genes in colon cancer stem-like cells including survivin, Bcl-XL, as well as Notch-1, -3, and -4, which may be involved in stem cell function. Furthermore, FLLL32 inhibited cell viability and tumorsphere formation as well as induced cleaved caspase-3 in colon cancer stem-like cells. FLLL32 is more potent than curcumin as evidenced with lower IC50 in colon cancer stem-like cells. In summary, our results indicate that STAT3 is a novel therapeutic target in colon cancer stem-like cells and inhibition of STAT3 in cancer stem-like cells may offer a potential treatment for colorectal cancer.

  5. Involvement of HIF-2?-mediated inflammation in arsenite-induced transformation of human bronchial epithelial cells

    SciTech Connect (OSTI)

    Xu, Yuan; Zhao, Yue; Xu, Wenchao; Luo, Fei; Wang, Bairu; Li, Yuan; Pang, Ying; Liu, Qizhan, E-mail: drqzliu@hotmail.com

    2013-10-15

    Arsenic is a well established human carcinogen that causes diseases of the lung. Some studies have suggested a link between inflammation and lung cancer; however, it is unknown if arsenite-induced inflammation causally contributes to arsenite-caused malignant transformation of cells. In this study, we investigated the molecular mechanisms underlying inflammation during neoplastic transformation induced in human bronchial epithelial (HBE) cells by chronic exposure to arsenite. The results showed that, on acute or chronic exposure to arsenite, HBE cells over-expressed the pro-inflammatory cytokines, interleukin-6 (IL-6), interleukin-8 (IL-8), and interleukin-1? (IL-1?). The data also indicated that HIF-2? was involved in arsenite-induced inflammation. Moreover, IL-6 and IL-8 were essential for the malignant progression of arsenite-transformed HBE cells. Thus, these experiments show that HIF-2? mediates arsenite-induced inflammation and that such inflammation is involved in arsenite-induced malignant transformation of HBE cells. The results provide a link between the inflammatory response and the acquisition of a malignant transformed phenotype by cells chronically exposed to arsenite and thus establish a previously unknown mechanism for arsenite-induced carcinogenesis. - Highlights: • Arsenite induces inflammation. • Arsenite-induced the increases of IL-6 and IL-8 via HIF-2?. • Inflammation is involved in arsenite-induced carcinogenesis.

  6. BDNF/TrkB signaling protects HT-29 human colon cancer cells from EGFR inhibition

    SciTech Connect (OSTI)

    Brunetto de Farias, Caroline; Children's Cancer Institute, 90420-140 Porto Alegre, RS; Laboratory of Neuropharmacology and Neural Tumor Biology, Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, 90050-170 Porto Alegre, RS; National Institute for Translational Medicine , 90035-003 Porto Alegre, RS ; Heinen, Tiago Elias; Pereira dos Santos, Rafael; Laboratory of Neuropharmacology and Neural Tumor Biology, Department of Pharmacology, Institute for Basic Health Sciences, Federal University of Rio Grande do Sul, 90050-170 Porto Alegre, RS; National Institute for Translational Medicine , 90035-003 Porto Alegre, RS ; Abujamra, Ana Lucia; Children's Cancer Institute, 90420-140 Porto Alegre, RS; National Institute for Translational Medicine , 90035-003 Porto Alegre, RS ; Schwartsmann, Gilberto; National Institute for Translational Medicine , 90035-003 Porto Alegre, RS; Department of Internal Medicine, School of Medicine, Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS ; and others

    2012-08-24

    Highlights: Black-Right-Pointing-Pointer BDNF protected HT-29 colorectal cancer cells from the antitumor effect of cetuximab. Black-Right-Pointing-Pointer TrkB inhibition potentiated the antitumor effect of cetuximab. Black-Right-Pointing-Pointer BDNF/TrkB signaling might be involved in resistance to anti-EGFR therapy. -- Abstract: The clinical success of targeted treatment of colorectal cancer (CRC) is often limited by resistance to anti-epidermal growth factor receptor (EGFR) therapy. The neurotrophin brain-derived neurotrophic factor (BDNF) and its receptor TrkB have recently emerged as anticancer targets, and we have previously shown increased BDNF levels in CRC tumor samples. Here we report the findings from in vitro experiments suggesting that BDNF/TrkB signaling can protect CRC cells from the antitumor effects of EGFR blockade. The anti-EGFR monoclonal antibody cetuximab reduced both cell proliferation and the mRNA expression of BDNF and TrkB in human HT-29 CRC cells. The inhibitory effect of cetuximab on cell proliferation and survival was counteracted by the addition of human recombinant BDNF. Finally, the Trk inhibitor K252a synergistically enhanced the effect of cetuximab on cell proliferation, and this effect was blocked by BDNF. These results provide the first evidence that increased BDNF/TrkB signaling might play a role in resistance to EGFR blockade. Moreover, it is possible that targeting TrkB could potentiate the anticancer effects of anti-EGFR therapy.

  7. Loss of p53 protein during radiation transformation of primary human mammary epithelial cells

    SciTech Connect (OSTI)

    Wazer, D.E.; Chu, Qiuming; Liu, Xiao Long; Gao, Qingshen; Safaii, H.; Band, V. (Tufts Univ. School of Medicine, Boston, MA (United States))

    1994-04-01

    The causative factors leading to breast cancer are largely unknown. Increased incidence of breast cancer following diagnostic or therapeutic radiation suggests that radiation may contribute to mammary oncogenesis. This report describes the in vitro neoplastic transformation of a normal human mammary epithelial cell strain, 76N, by fractionated [gamma]-irradiation at a clinically used dose (30 Gy). The transformed cells (76R-30) were immortal, had reduced growth factor requirements, and produced tumors in nude mice. Remarkably, the 76R-30 cells completely lacked the p53 tumor suppressor protein. Loss of p53 was due to deletion of the gene on one allele and a 26-bp deletion within the third intron on the second allele which resulted in abnormal splicing out of either the third or fourth exon from the mRNA. PCR with a mutation-specific primer showed that intron 3 mutation was present in irradiated cells before selection for immortal phenotype. 76R-30 cells did not exhibit G[sub 1] arrest in response to radiation, indicating a loss of p53-mediated function. Expression of the wild-type p53 gene in 76R-30 cells led to their growth inhibition. Thus, loss of p53 protein appears to have contributed to neoplastic transformation of these cells. This unique model should facilitate analyses of molecular mechanisms of radiation-induced breast cancer and allow identification of p53-regulated cellular genes in breast cells. 44 refs., 8 figs., 1 tab.

  8. Mueller matrix imaging for skin cancer detection 

    E-Print Network [OSTI]

    Baldwin, Angela Michelle

    2004-09-30

    is suspect, a biopsy is performed to verify the diagnosis. For squamous cell and basal cell carcinomas, the most common treatment options are excision, curettage and electrodesiccation, cryosurgery, radiation, and Moh?s micrographic surgery. Excision... is where all visible cancer is cut away together with a 3 to 10 mm margin of healthy tissue and then the skin is stitched closed with sutures. Curettage and electrodesiccation are where all visible cancer is scraped away and then an electric probe...

  9. CDK-associated Cullin 1 promotes cell proliferation with activation of ERK1/2 in human lung cancer A549 cells

    SciTech Connect (OSTI)

    Chen, Tian Jun; Gao, Fei; Yang, Tian; Thakur, Asmitanand; Ren, Hui; Li, Yang; Zhang, Shuo; Wang, Ting; Chen, Ming Wei

    2013-07-19

    Highlights: •CDK-associated Cullin 1 (CAC1) expression increases in human lung carcinoma. •CAC1 promotes the proliferation of lung cancer A549 cells. •CAC1 promotes human lung cancer A549 cell proliferation with activation of ERK1/2. -- Abstract: Lung cancer is one of the most common causes of cancer-related death in the world, but the mechanisms remain unknown. In this study, we investigated the expression of CDK-associated Cullin 1 (CAC1) in lung cancer, the effect of CAC1 on the proliferation of human lung cancer A549 cells, and the activation of signaling pathways of mitogen-activated protein kinases (MAPKs). Results showed that CAC1 expression was higher levels in human lung carcinoma than normal lung tissue, and CAC1 siRNA reduced the proliferation of lung cancer A549 cells by decreasing cell activity and cell division in vitro. The proportion of cells treated with CAC1 siRNA increased in the G1 phase and decreased in the S and G2/M phase, indicative of G1 cell cycle arrest. Furthermore, the proportions of early/late apoptosis in lung cancer A549 cells were enhanced with CAC1 siRNA treatment. It was also found that activation of extracellular signal-regulated protein kinase (ERK) and p38 signaling pathways were involved in the proliferation of A549 cells. After CAC1 siRNA treatment, p-ERK1/2 levels decreased, and meanwhile p-p38 level increased, A549 cell proliferation increased when ERK1/2 signaling is activated by PMA. Our findings demonstrated that CAC1 promoted the proliferation of human lung cancer A549 cells with activation of ERK1/2 signaling pathways, suggesting a potential cure target for treatment of human lung cancer.

  10. Soft inertial microfluidics for high throughput separation of bacteria from human blood cells

    SciTech Connect (OSTI)

    Wu, Zhigang; Willing, Ben; Bjerketorp, Joakim; Jansson, Janet K.; Hjort, Klas

    2009-01-05

    We developed a new approach to separate bacteria from human blood cells based on soft inertial force induced migration with flow defined curved and focused sample flow inside a microfluidic device. This approach relies on a combination of an asymmetrical sheath flow and proper channel geometry to generate a soft inertial force on the sample fluid in the curved and focused sample flow segment to deflect larger particles away while the smaller ones are kept on or near the original flow streamline. The curved and focused sample flow and inertial effect were visualized and verified using a fluorescent dye primed in the device. First the particle behavior was studied in detail using 9.9 and 1.0 {micro}m particles with a polymer-based prototype. The prototype device is compact with an active size of 3 mm{sup 2}. The soft inertial effect and deflection distance were proportional to the fluid Reynolds number (Re) and particle Reynolds number (Re{sub p}), respectively. We successfully demonstrated separation of bacteria (Escherichia coli) from human red blood cells at high cell concentrations (above 10{sup 8}/mL), using a sample flow rate of up to 18 {micro}L/min. This resulted in at least a 300-fold enrichment of bacteria at a wide range of flow rates with a controlled flow spreading. The separated cells were proven to be viable. Proteins from fractions before and after cell separation were analyzed by gel electrophoresis and staining to verify the removal of red blood cell proteins from the bacterial cell fraction. This novel microfluidic process is robust, reproducible, simple to perform, and has a high throughput compared to other cell sorting systems. Microfluidic systems based on these principles could easily be manufactured for clinical laboratory and biomedical applications.

  11. Transcription factor co-localization patterns affect human cell type-specific gene expression.

    E-Print Network [OSTI]

    Wang, Dennis; Rendon, Augusto; Ouwehand, Willem; Wernisch, Lorenz

    2012-06-21

    Wang et al. BMC Genomics 2012, 13:263 http://www.biomedcentral.com/1471-2164/13/263 RESEARCH ARTICLE Open Access Transcription factor co-localization patterns affect human cell type-specific gene expression Dennis Wang1*, Augusto Rendon2, Willem... identified thousands of regions along the *Correspondence: dennis.wang@mrc-bsu.cam.ac.uk 1MRC Biostatistics Unit, Institute of Public Health, Robinson Way, Cambridge, UK Full list of author information is available at the end of the article human genome where...

  12. Human Haploid Cell Genetics Reveals Roles for Lipid Metabolism Genes in Nonapoptotic Cell Death

    E-Print Network [OSTI]

    Stockwell, Brent R.

    Corner Building, MC 4846, New York, New York 10027, United States CeMM Research Center for Molecular, Stanford University, 337 Campus Drive, Stanford, California 94305, United States *S Supporting Information) in ferroptosis. One novel compound, CIL56, triggered cell death dependent upon the rate-limiting de novo lipid

  13. The pyrimidine nucleotide carrier PNC1 and mitochondrial trafficking of thymidine phosphates in cultured human cells

    SciTech Connect (OSTI)

    Franzolin, Elisa; Miazzi, Cristina; Frangini, Miriam; Palumbo, Elisa; Rampazzo, Chiara [Department of Biology, University of Padova, Via Ugo Bassi 58B, I-35131 Padova (Italy)] [Department of Biology, University of Padova, Via Ugo Bassi 58B, I-35131 Padova (Italy); Bianchi, Vera, E-mail: vbianchi@bio.unipd.it [Department of Biology, University of Padova, Via Ugo Bassi 58B, I-35131 Padova (Italy)] [Department of Biology, University of Padova, Via Ugo Bassi 58B, I-35131 Padova (Italy)

    2012-10-15

    In cycling cells cytosolic de novo synthesis of deoxynucleotides is the main source of precursors for mitochondrial (mt) DNA synthesis. The transfer of deoxynucleotides across the inner mt membrane requires protein carriers. PNC1, a SLC25 family member, exchanges pyrimidine nucleoside triphosphates in liposomes and its downregulation decreases mtUTP concentration in cultured cells. By an isotope-flow protocol we confirmed transport of uridine nucleotides by PNC1 in intact cultured cells and investigated PNC1 involvement in the mt trafficking of thymidine phosphates. Key features of our approach were the manipulation of PNC1 expression by RNA interference or inducible overexpression, the employment of cells proficient or deficient for cytosolic thymidine kinase (TK1) to distinguish the direction of flow of thymidine nucleotides across the mt membrane during short pulses with [{sup 3}H]-thymidine, the determination of mtdTTP specific radioactivity to quantitate the rate of mtdTTP export to the cytoplasm. Downregulation of PNC1 in TK1{sup -} cells increased labeled dTTP in mitochondria due to a reduced rate of export. Overexpression of PNC1 in TK1{sup +} cells increased mtdTTP pool size and radioactivity, suggesting an involvement in the import of thymidine phosphates. Thus PNC1 is a component of the network regulating the mtdTTP pool in human cells. -- Highlights: Black-Right-Pointing-Pointer Thymidine phosphates exchange between mitochondria and cytosol in mammalian cells. Black-Right-Pointing-Pointer siRNA-downregulation of PNC1 delays mitochondrial dTTP export in TK1{sup -} cells. Black-Right-Pointing-Pointer PNC1 overexpression accumulates dTTP in mitochondria of TK1{sup +} cells. Black-Right-Pointing-Pointer PNC1 exchanges thymidine nucleotides across the mitochondrial inner membrane. Black-Right-Pointing-Pointer PNC1 participates in the regulation of the mtdTTP pool supporting mtDNA synthesis.

  14. Theophylline prevents NAD{sup +} depletion via PARP-1 inhibition in human pulmonary epithelial cells

    SciTech Connect (OSTI)

    Moonen, Harald J.J. . E-mail: h.moonen@grat.unimaas.nl; Geraets, Liesbeth; Vaarhorst, Anika; Bast, Aalt; Wouters, Emiel F.M.; Hageman, Geja J.

    2005-12-30

    Oxidative DNA damage, as occurs during exacerbations in chronic obstructive pulmonary disease (COPD), highly activates the nuclear enzyme poly(ADP-ribose)polymerase-1 (PARP-1). This can lead to cellular depletion of its substrate NAD{sup +}, resulting in an energy crisis and ultimately in cell death. Inhibition of PARP-1 results in preservation of the intracellular NAD{sup +} pool, and of NAD{sup +}-dependent cellular processes. In this study, PARP-1 activation by hydrogen peroxide decreased intracellular NAD{sup +} levels in human pulmonary epithelial cells, which was found to be prevented in a dose-dependent manner by theophylline, a widely used compound in the treatment of COPD. This enzyme inhibition by theophylline was confirmed in an ELISA using purified human PARP-1 and was found to be competitive by nature. These findings provide new mechanistic insights into the therapeutic effect of theophylline in oxidative stress-induced lung pathologies.

  15. Live-cell imaging RNAi screen identifies PP2AB55 and importin-1 as key mitotic exit regulators in human cells

    E-Print Network [OSTI]

    Trinkle-Mulcahy, Laura

    subunits that associate with a small group of catalytic subunits3,4,17 . It is possible that mitotic exit mitotic exit in live human cells, we measured the timing from anaphase onset until nuclear reformation. We

  16. VI-14, a novel flavonoid derivative, inhibits migration and invasion of human breast cancer cells

    SciTech Connect (OSTI)

    Li, Fanni; Li, Chenglin; Zhang, Haiwei; Lu, Zhijian [State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009 (China)] [State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009 (China); Li, Zhiyu; You, Qidong [Department of Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009 (China)] [Department of Medicinal Chemistry, China Pharmaceutical University, Nanjing 210009 (China); Lu, Na [State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009 (China)] [State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009 (China); Guo, Qinglong, E-mail: anticancer_drug@yahoo.com.cn [State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009 (China)] [State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, 24 Tongjiaxiang, Nanjing 210009 (China)

    2012-06-01

    It has been well characterized that flavonoids possess pronounced anticancer potentials including anti-angiogenesis, anti-metastasis, and pro-apoptosis. Herein, we report, for the first time, that VI-14, a novel flavonoid derivative, possesses anti-cancer properties. The purpose of this study is to investigate the anti-migration and anti-invasion activities of VI-14 in breast cancer cells. Our data indicate that VI-14 inhibits adhesion, migration and invasion of MDA-MB-231 and MDA-MB-435 human breast cancer cells. MDA-MB-231 cells treated with VI-14 display reduced activities and expressions of ECM degradation-associated proteins including matrix metalloproteinase 2 (MMP-2) and 9 (MMP-9) at both the protein and mRNA levels. Meanwhile, VI-14 treatment induces an up-regulated expression of tissue inhibitor of metalloproteinase 1 (TIMP-1) and 2 (TIMP-2) in MDA-MB-231 cells. Western blotting results show that phosphorylation levels of critical components of the MAPK signaling pathway, including ERK, JNK and P38, are dramatically decreased in VI-14-treated MDA-MB-231 cells. Furthermore, treatment of VI-14 significantly decreases the nuclear levels and the binding ability of nuclear factor-kappa B (NF-?B) and activator protein-1 (AP-1). Taken together, our data suggest that VI-14 treatment suppresses migration and motility of breast cancer cells, and VI-14 may be a potential compound for cancer therapy. Highlights: ? We report for the first time that VI-14 possesses anti-cancer properties. ? VI-14 weakens the adhesion, migration and invasion of human breast cancer cells. ? VI-14 decreases the activities and expressions of MMP-2/9. ? VI-14 suppresses the phosphorylation levels of the MAPK signaling pathway. ? VI-14 decreases the nuclear levels and the binding ability of NF-?B and AP-1.

  17. Sprayed skin turbine component

    DOE Patents [OSTI]

    Allen, David B

    2013-06-04

    Fabricating a turbine component (50) by casting a core structure (30), forming an array of pits (24) in an outer surface (32) of the core structure, depositing a transient liquid phase (TLP) material (40) on the outer surface of the core structure, the TLP containing a melting-point depressant, depositing a skin (42) on the outer surface of the core structure over the TLP material, and heating the assembly, thus forming both a diffusion bond and a mechanical interlock between the skin and the core structure. The heating diffuses the melting-point depressant away from the interface. Subsurface cooling channels (35) may be formed by forming grooves (34) in the outer surface of the core structure, filling the grooves with a fugitive filler (36), depositing and bonding the skin (42), then removing the fugitive material.

  18. Noscapine induces mitochondria-mediated apoptosis in human colon cancer cells in vivo and in vitro

    SciTech Connect (OSTI)

    Yang, Zi-Rong; Liu, Meng; Peng, Xiu-Lan; Lei, Xiao-Fei; Zhang, Ji-Xiang; Dong, Wei-Guo

    2012-05-11

    Highlights: Black-Right-Pointing-Pointer Noscapine inhibited cell viability of colon cancer in a time- and dose- dependent manner. Black-Right-Pointing-Pointer G{sub 2}/M phase arrest and chromatin condensation and nuclear fragmentation were induced. Black-Right-Pointing-Pointer Noscapine promoted apoptosis via mitochondrial pathways. Black-Right-Pointing-Pointer Tumorigenicity was inhibited by noscapine. -- Abstract: Noscapine, a phthalide isoquinoline alkaloid derived from opium, has been widely used as a cough suppressant for decades. Noscapine has recently been shown to potentiate the anti-cancer effects of several therapies by inducing apoptosis in various malignant cells without any detectable toxicity in cells or tissues. However, the mechanism by which noscapine induces apoptosis in colon cancer cells remains unclear. The signaling pathways by which noscapine induces apoptosis were investigated in colon cancer cell lines treated with various noscapine concentrations for 72 h, and a dose-dependent inhibition of cell viability was observed. Noscapine effectively inhibited the proliferation of LoVo cells in vitro (IC{sub 50} = 75 {mu}M). This cytotoxicity was reflected by cell cycle arrest at G{sub 2}/M and subsequent apoptosis, as indicated by increased chromatin condensation and fragmentation, the upregulation of Bax and cytochrome c (Cyt-c), the downregulation of survivin and Bcl-2, and the activation of caspase-3 and caspase-9. Moreover, in a xenograft tumor model in mice, noscapine injection clearly inhibited tumor growth via the induction of apoptosis, which was demonstrated using a TUNEL assay. These results suggest that noscapine induces apoptosis in colon cancer cells via mitochondrial pathways. Noscapine may be a safe and effective chemotherapeutic agent for the treatment of human colon cancer.

  19. Disrupting the wall accumulation of human sperm cells by artificial corrugation

    E-Print Network [OSTI]

    H. A. Guidobaldi; Y. Jeyaram; C. A. Condat; M. Oviedo; I. Berdakin; V. V. Moshchalkov; L. C. Giojalas; A. V. Silhanek; V. I. Marconi

    2015-04-29

    Many self-propelled microorganisms are attracted to surfaces. This makes their dynamics in restricted geometries very different from that observed in the bulk. Swimming along walls is beneficial for directing and sorting cells, but may be detrimental if homogeneous populations are desired, such as in counting microchambers. In this work, we characterize the motion of human sperm cells $60 \\mu m$ long, strongly confined to $25 \\mu m$ shallow chambers. We investigate the nature of the cell trajectories between the confining surfaces and their accumulation near the borders. Observed cell trajectories are composed of a succession of quasi-circular and quasi-linear segments. This suggests that the cells follow a path of intermittent trappings near the top and bottom surfaces separated by stretches of quasi-free motion in between the two surfaces, as confirmed by depth resolved confocal microscopy studies. We show that the introduction of artificial petal-shaped corrugation in the lateral boundaries removes the tendency of cells to accumulate near the borders, an effect which we hypothesize may be valuable for microfluidic applications in biomedicine.

  20. Chip-Based Comparison of the Osteogenesis of Human Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem Cells under Mechanical Stimulation

    E-Print Network [OSTI]

    Park, Sang-Hyug

    Adipose tissue-derived stem cells (ASCs) are considered as an attractive stem cell source for tissue engineering and regenerative medicine. We compared human bone marrow-derived mesenchymal stem cells (hMSCs) and hASCs ...

  1. HER/ErbB Receptor Interactions and Signaling Patterns in Human Mammary Epithelial Cells

    SciTech Connect (OSTI)

    Zhang, Yi; Opresko, Lee K.; Shankaran, Harish; Chrisler, William B.; Wiley, H. S.; Resat, Haluk

    2009-10-31

    Knowledge about signaling pathways is typically compiled based on data gathered using different cell lines. This approach implicitly assumes that cell line dependence is not important, which can be misleading because different cell lines do not always respond to a particular stimulus in the same way. The lack of coherent data collected from closely related cellular systems can be detrimental to the efforts to understand the regulation of biological processes. In this study, we report the development of a library of human mammary epithelial (HME) cell lines which express endogenous levels of the cell surface receptor EGFR/HER1, and different levels of HER2 and HER3. Using our clone library, we have quantified the interactions among the HER1-3 receptors and systematically investigated the existing hypotheses about their interaction patterns. Contrary to earlier suggestions, we find that lateral interactions with HER2 do not lead to strong transactivation between EGFR and HER3. Our study identified HER2 as the dominant dimerization partner for both EGFR and HER3, and revealed that EGFR and HER3 activations are only weakly linked in HME cells. We have also quantified the time-dependent activation patterns of the downstream effectors Erk and Akt. We found that HER3 signaling makes the strongest contribution to Akt activation and that, stimulation of either EGFR or HER3 pathways activate Erk at significant levels. Our study shows that cell libraries formed from closely related clones can be a powerful resource for pursuing the quantitative investigations that are necessary for developing a systems level understanding of cell signaling.

  2. A Comparative Study on Human Embryonic Stem Cell Patent Law in the United States, the European Patent Organization, and China

    E-Print Network [OSTI]

    Zhu, Huan

    2011-05-31

    With the recent developments in biotechnology, associated patent law issues have been a growing concern since the 1980s. Among all the subcategories within the general field of biotechnology, human embryonic stem cell ...

  3. Novel Direct Targets and Functional Roles for MicroRNA-21 in Granulosa Cells and Human Uterine Leiomyomas

    E-Print Network [OSTI]

    Fitzgerald, Jonathan Browning

    2013-08-31

    MicroRNA-21 (miR-21) is important for maintaining optimal ovulation rates and granulosa cell viability. It is also upregulated in human uterine leiomyomas (ULMs), a disease characterized by the presence of benign tumors ...

  4. Chondrogenesis and mineralization during in vitro culture of human stem cells on 3D-woven scaffolds

    E-Print Network [OSTI]

    Abrahamsson, Christoffer K.

    Human mesenchymal stem cells (hMSCs) and three-dimensional (3D) woven poly(?-caprolactone) (PCL) scaffolds are promising tools for skeletal tissue engineering. We hypothesized that in vitro culture duration and medium ...

  5. Comparative Analysis of Live, Heat-inactivated, and Electron Beam Inactivated Salmonella Typhimurium Infection in Human Host Cells 

    E-Print Network [OSTI]

    Corkill, Carolina

    2013-08-07

    Salmonella Typhimurium continues to be a leading cause of human gastroenteritis worldwide. This organism is a facultative intracellular pathogen, meaning that it is able grow and reproduce within the host cell it inhabits. S...

  6. VEGF induces proliferation of human hair follicle dermal papilla cells through VEGFR-2-mediated activation of ERK

    SciTech Connect (OSTI)

    Li, Wei; Man, Xiao-Yong [Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009 (China)] [Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009 (China); Li, Chun-Ming [Department of Dermatology, Second Affiliated Hospital, Nanchang University School of Medicine, Nanchang, Jiangxi 330000 (China)] [Department of Dermatology, Second Affiliated Hospital, Nanchang University School of Medicine, Nanchang, Jiangxi 330000 (China); Chen, Jia-Qi; Zhou, Jiong; Cai, Sui-Qing; Lu, Zhong-Fa [Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009 (China)] [Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009 (China); Zheng, Min, E-mail: minz@zju.edu.cn [Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009 (China)] [Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009 (China)

    2012-08-15

    Vascular endothelial growth factor (VEGF) is one of the strongest regulators of physiological and pathological angiogenesis. VEGF receptor 2 (VEGFR-2), the primary receptor for VEGF, is thought to mediate major functional effects of VEGF. Previously, we have localized both VEGF and VEGFR-2 in human hair follicles. In this study, we further defined the expression and roles of VEGFR-2 on human hair follicle dermal papilla (DP) cells. The expression of VEGFR-2 on DP cells was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis separately, and localization of VEGFR-2 was defined by immunofluorescence. The effect of VEGF on DP cells was analyzed by MTT assays and specific inhibitors. Finally, the role of VEGF involved in the signaling pathways was investigated by Western blot. RT-PCR and Western blot analysis demonstrated the expression of VEGFR-2 on DP cells. Immunostaining for VEGFR-2 showed strong signal on cultured human DP cells in vitro. Exogenous VEGF{sub 165} stimulated proliferation of DP cells in a dose-dependent manner. Furthermore, this stimulation was blocked by a VEGFR-2 neutralizing antibody (MAB3571) and an ERK inhibitor (PD98059). VEGF{sub 165}-induced phosphorylation of ERK1/2 was abolished by MAB3571 and PD98059, while the phosphorylation of p38, JNK and AKT were not changed by VEGF{sub 165}. Taken together, VEGFR-2 is expressed on primary human hair follicle DP cells and VEGF induces proliferation of DP cells through VEGFR-2/ERK pathway, but not p38, JNK or AKT signaling. -- Highlights: Black-Right-Pointing-Pointer We examine the expression of VEGFR-2 on cultured human dermal papilla (DP) cells. Black-Right-Pointing-Pointer VEGF{sub 165} stimulated proliferation of human DP cells in a dose-dependent manner. Black-Right-Pointing-Pointer This stimulation was through VEGFR-2-mediated activation of ERK.

  7. Human T-lymphotropic Virus Type 1-infected Cells Secrete Exosomes That Contain Tax Protein*S

    E-Print Network [OSTI]

    Vertes, Akos

    Human T-lymphotropic Virus Type 1-infected Cells Secrete Exosomes That Contain Tax Protein, the ¶ Unité d'Epidémiologie et Physiopathologie des Virus Oncogènes, Département de Virologie, Institut, or tropical spastic paraparesis. Human T-lymphotropic virus type 1 (HTLV-1) is the causa- tive agent of adult

  8. Activation of ERK and JNK signaling pathways by mycotoxin citrinin in human cells

    SciTech Connect (OSTI)

    Chang, C.-H.; Yu, F.-Y.; Wang, L.-T.; Lin, Y.-S. [Department of Biomedical Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Rd., Taichung 402, Taiwan (China); Liu, B.-H. [Department of Biomedical Sciences, Chung Shan Medical University, No. 110, Sec. 1, Chien-Kuo N. Rd., Taichung 402, Taiwan (China)], E-mail: bingliu@csmu.edu.tw

    2009-06-15

    Mycotoxin citrinin (CTN) is commonly found in foods and feeds that are contaminated/inoculated with Penicillium, Aspergillus and Monascus species. The exposure of human embryonic kidney (HEK293) and HeLa cells to CTN resulted in a dose-dependent increase in the phosphorylation of two major mitogen-activated protein kinases (MAPKs), ERK1/2 and JNK. In HEK293 cultures, the administering of CTN increased both the mRNA and protein levels of egr-1, c-fos and c-jun genes; additionally, the ERK1/2 pathway contributed to the upregulation of Egr-1 and c-Fos protein expression. CTN treatment also induced the transcription activity of Egr-1 and AP-1 proteins, as evidenced by luciferase reporter assays. Bioinformatic analyses indicated two genes Gadd45{beta} and MMP3 have Egr-1 and AP-1 response elements in their promoters, respectively. Furthermore, co-exposure of HEK293 cells to CTN and MAPK pathway inhibitors demonstrated that CTN increased the levels of Gadd45{beta} mRNA through ERK1/2 signaling pathway and up-regulated the MMP3 transcripts majorly via JNK pathway. Finally, CTN-triggered caspase 3 activity was significantly reduced in the presence of MAPK inhibitors. Our results suggest that CTN positively regulates ERK1/2 and JNK pathways as well as their downstream effectors in human cells; activated MAPK pathways are also involved in CTN-induced apoptosis.

  9. Induction of human breast cell carcinogenesis by triclocarban and intervention by curcumin

    SciTech Connect (OSTI)

    Sood, Shilpa; Choudhary, Shambhunath; Wang, Hwa-Chain Robert

    2013-09-06

    Highlights: •Triclocarban exposure induces breast epithelial cell carcinogenesis. •Triclocarban induces the Erk–Nox pathway, ROS elevation, and DNA damage. •Physiological doses of triclocarban induce cellular carcinogenesis. •Non-cytotoxic curcumin blocks triclocarban-induced carcinogenesis and pathways. -- Abstract: More than 85% of breast cancers are sporadic and attributable to long-term exposure to environmental carcinogens and co-carcinogens. To identify co-carcinogens with abilities to induce cellular pre-malignancy, we studied the activity of triclocarban (TCC), an antimicrobial agent commonly used in household and personal care products. Here, we demonstrated, for the first time, that chronic exposure to TCC at physiologically-achievable nanomolar concentrations resulted in progressive carcinogenesis of human breast cells from non-cancerous to pre-malignant. Pre-malignant carcinogenesis was measured by increasingly-acquired cancer-associated properties of reduced dependence on growth factors, anchorage-independent growth and increased cell proliferation, without acquisition of cellular tumorigenicity. Long-term TCC exposure also induced constitutive activation of the Erk–Nox pathway and increases of reactive oxygen species (ROS) in cells. A single TCC exposure induced transient induction of the Erk–Nox pathway, ROS elevation, increased cell proliferation, and DNA damage in not only non-cancerous breast cells but also breast cancer cells. Using these constitutively- and transiently-induced changes as endpoints, we revealed that non-cytotoxic curcumin was effective in intervention of TCC-induced cellular pre-malignancy. Our results lead us to suggest that the co-carcinogenic potential of TCC should be seriously considered in epidemiological studies to reveal the significance of TCC in the development of sporadic breast cancer. Using TCC-induced transient and constitutive endpoints as targets will likely help identify non-cytotoxic preventive agents, such as curcumin, effective in suppressing TCC-induced cellular pre-malignancy.

  10. Biochemical signatures of in vitro radiation response in human lung, breast and prostate tumour cells observed with Raman spectroscopy

    E-Print Network [OSTI]

    Brolo, Alexandre G.

    cells observed with Raman spectroscopy This article has been downloaded from IOPscience. Please scroll down to see the full text article. 2011 Phys. Med. Biol. 56 6839 (http://iopscience.iop.org/0031 signatures of in vitro radiation response in human lung, breast and prostate tumour cells observed with Raman

  11. Skin contamination dosimeter

    DOE Patents [OSTI]

    Hamby, David M. (Corvallis, OR); Farsoni, Abdollah T. (Corvallis, OR); Cazalas, Edward (Corvallis, OR)

    2011-06-21

    A technique and device provides absolute skin dosimetry in real time at multiple tissue depths simultaneously. The device uses a phoswich detector which has multiple scintillators embedded at different depths within a non-scintillating material. A digital pulse processor connected to the phoswich detector measures a differential distribution (dN/dH) of count rate N as function of pulse height H for signals from each of the multiple scintillators. A digital processor computes in real time from the differential count-rate distribution for each of multiple scintillators an estimate of an ionizing radiation dose delivered to each of multiple depths of skin tissue corresponding to the multiple scintillators embedded at multiple corresponding depths within the non-scintillating material.

  12. Long term culture of genome-stable bipotent progenitor cells from adult human liver

    E-Print Network [OSTI]

    Huch, Meritxell; Gehart, Helmuth; van Boxtel, Ruben; Hamer, Karien; Blokzijl, Francis; Verstegen, Monique M. A.; Ellis, Ewa; van Wenum, Martien; Fuchs, Sabine A.; de Ligt, Joep; van de Wetering, Marc; Sasaki, Nobuo; Boers, Susanne J.; Kemperman, Hans; de Jonge, Jeroen; Ijzermans, Jan N. M.; Niewenhuis, Edward; Hoekstra, Ruurdtje; Strom, Stephen; Vries, Robert R. G.; van der Laan, Luc J. W.; Cuppen, Edwin; Clevers, Hans

    2014-12-18

    , S.K., Larson, J.J., Yawn, B., Therneau, T.M., and Kim, W.R. (2013). Underestimation of liver-related mortality in the United States. Gastroenterology 145, 375- 382 e371-372. Baker, D.E., Harrison, N.J., Maltby, E., Smith, K., Moore, H.D., Shaw, P... .J., Heath, P.R., Holden, H., and Andrews, P.W. (2007). Adaptation to culture of human embryonic stem cells and oncogenesis in vivo. Nat Biotechnol 25, 207-215. Barker, N., Huch, M., Kujala, P., van de Wetering, M., Snippert, H.J., van Es, J.H., Sato, T...

  13. Cell cycle regulation of human immunodeficiency virus type 1 integration in T cells: antagonistic effects of nuclear envelope breakdown and chromatin condensation

    SciTech Connect (OSTI)

    Mannioui, Abdelkrim . E-mail: karim.mannioui@chu-stlouis.fr; Schiffer, Cecile . E-mail: cecile.schiffer@voila.fr; Felix, Nathalie . E-mail: nathalie.felix@chu-stlouis.fr

    2004-11-10

    We examined the influence of mitosis on the kinetics of human immunodeficiency virus type 1 integration in T cells. Single-round infection of cells arrested in G1b or allowed to synchronously proceed through division showed that mitosis delays virus integration until 18-24 h postinfection, whereas integration reaches maximum levels by 15 h in G1b-arrested cells. Subcellular fractionation of metaphase-arrested cells indicated that, while nuclear envelope disassembly facilitates docking of viral DNA to chromatin, chromosome condensation directly antagonizes and therefore delays integration. As a result of the balance between the two effects, virus integration efficiency is eventually up to threefold greater in dividing cells. At the single-cell level, using a green fluorescent protein-expressing reporter virus, we found that passage through mitosis leads to prominent asymmetric segregation of the viral genome in daughter cells without interfering with provirus expression.

  14. Effects of ultrasound and sodium lauryl sulfate on the transdermal delivery of hydrophilic permeants: Comparative in vitro studies with full-thickness and split-thickness pig and human skin

    E-Print Network [OSTI]

    Seto, Jennifer E.

    The simultaneous application of ultrasound and the surfactant sodium lauryl sulfate (referred to as US/SLS) to skin enhances transdermal drug delivery (TDD) in a synergistic mechanical and chemical manner. Since full-thickness ...

  15. Viability of adult rat skin following 13 Mev proton irradiation 

    E-Print Network [OSTI]

    Caraway, Bobby Lamar

    1966-01-01

    alteration resulting from total skin proton irradiation seemed to be dose related. Therefore, since the amount of tissue alteration seems to be dose-dependent, a hypothesis was developed that growth and viability of skin cells removed' by biopsy... rats each were subjected to total-skin proton irradiation of varying doses. The dose varied from 1300 rad in Group I to 200 rad in Group 1V. Two rats from each group served as controls and received no irradiation. Five days and 30 days...

  16. Nicotine induces fibrogenic changes in human liver via nicotinic acetylcholine receptors expressed on hepatic stellate cells

    SciTech Connect (OSTI)

    Soeda, Junpei; Morgan, Maelle; McKee, Chad; Mouralidarane, Angelina; Lin, ChingI [University College London, Centre for Hepatology, Royal Free Hospital, London NW3 2PF (United Kingdom)] [University College London, Centre for Hepatology, Royal Free Hospital, London NW3 2PF (United Kingdom); Roskams, Tania [Department of Morphology and Molecular Pathology, University of Leuven (Belgium)] [Department of Morphology and Molecular Pathology, University of Leuven (Belgium); Oben, Jude A., E-mail: j.oben@ucl.ac.uk [University College London, Centre for Hepatology, Royal Free Hospital, London NW3 2PF (United Kingdom); Department of Gastroenterology and Hepatology, Guy's and St Thomas' Hospital, London SE1 7EH (United Kingdom)

    2012-01-06

    Highlights: Black-Right-Pointing-Pointer Cigarette smoke may induce liver fibrosis via nicotine receptors. Black-Right-Pointing-Pointer Nicotine induces proliferation of hepatic stellate cells (HSCs). Black-Right-Pointing-Pointer Nicotine activates hepatic fibrogenic pathways. Black-Right-Pointing-Pointer Nicotine receptor antagonists attenuate HSC proliferation. Black-Right-Pointing-Pointer Nicotinic receptor antagonists may have utility as novel anti-fibrotic agents. -- Abstract: Background and aims: Cigarette smoke (CS) may cause liver fibrosis but possible involved mechanisms are unclear. Among the many chemicals in CS is nicotine - which affects cells through nicotinic acetylcholine receptors (nAChR). We studied the effects of nicotine, and involved pathways, on human primary hepatic stellate cells (hHSCs), the principal fibrogenic cells in the liver. We then determined possible disease relevance by assaying nAChR in liver samples from human non-alcoholic steatohepatitis (NASH). Methods: hHSC were isolated from healthy human livers and nAChR expression analyzed - RT-PCR and Western blotting. Nicotine induction of hHSC proliferation, upregulation of collagen1-{alpha}2 and the pro-fibrogenic cytokine transforming growth factor beta 1 (TGF-{beta}1) was determined along with involved intracellular signaling pathways. nAChR mRNA expression was finally analyzed in whole liver biopsies obtained from patients diagnosed with non-alcoholic steatohepatitis (NASH). Results: hHSCs express muscle type ({alpha}1, {beta}1, delta and epsilon) and neuronal type ({alpha}3, {alpha}6, {alpha}7, {beta}2 and {beta}4) nAChR subunits at the mRNA level. Among these subunits, {alpha}3, {alpha}7, {beta}1 and {epsilon} were predominantly expressed as confirmed by Western blotting. Nicotine induced hHSC proliferation was attenuated by mecamylamine (p < 0.05). Additionally, collagen1-{alpha}2 and TGF-{beta}1 mRNA expression were significantly upregulated by nicotine and inhibited by mecamylamine. {alpha}1 and {alpha}3-nAChR mRNA expression was significantly upregulated in NASH fibrosis compared to normal livers. Conclusion: Nicotine at levels in smokers' blood is pro-fibrogenic, through actions on hHSCs expressed nAChRs. Therefore, CS, via its nicotine content, may worsen liver fibrosis. Moreover, nicotinic receptor antagonists may have utility as novel anti-fibrotic agents.

  17. CXCR6, a Newly Defined Biomarker of Tissue-Specific Stem Cell Asymmetric Self-Renewal, Identifies More Aggressive Human Melanoma Cancer Stem Cells

    E-Print Network [OSTI]

    Rouzbeh Taghizadeh; Minsoo Noh; Yang Hoon Huh; Emilio Ciusani; Luca Sigalotti; Michele Maio; Beatrice Arosio; Maria R. Nicotra; PierGiorgio Natali; James L. Sherley; Caterina A. M. La Porta

    2013-08-28

    Background: A fundamental problem in cancer research is identifying the cell type that is capable of sustaining neoplastic growth and its origin from normal tissue cells. Recent investigations of a variety of tumor types have shown that phenotypically identifiable and isolable subfractions of cells possess the tumor-forming ability. In the present paper, using two lineage-related human melanoma cell lines, primary melanoma line IGR39 and its metastatic derivative line IGR37, two main observations are reported. The first one is the first phenotypic evidence to support the origin of melanoma cancer stem cells (CSCs) from mutated tissue-specific stem cells; and the second one is the identification of a more aggressive subpopulation of CSCs in melanoma that are CXCR6+. Conclusions/Significance: The association of a more aggressive tumor phenotype with asymmetric self-renewal phenotype reveals a previously unrecognized aspect of tumor cell physiology. Namely, the retention of some tissue-specific stem cell attributes, like the ability to asymmetrically self-renew, impacts the natural history of human tumor development. Knowledge of this new aspect of tumor development and progression may provide new targets for cancer prevention and treatment.

  18. Zinc chromate induces chromosome instability and DNA double strand breaks in human lung cells

    SciTech Connect (OSTI)

    Xie Hong; Holmes, Amie L.; Young, Jamie L.; Qin Qin; Joyce, Kellie; Pelsue, Stephen C.; Peng Cheng; Wise, Sandra S.; Jeevarajan, Antony S.; Wallace, William T.; Hammond, Dianne; Wise, John Pierce E-mail: John.Wise@usm.maine.edu

    2009-02-01

    Hexavalent chromium Cr(VI) is a respiratory toxicant and carcinogen, with solubility playing an important role in its carcinogenic potential. Zinc chromate, a water insoluble or 'particulate' Cr(VI) compound, has been shown to be carcinogenic in epidemiology studies and to induce tumors in experimental animals, but its genotoxicity is poorly understood. Our study shows that zinc chromate induced concentration-dependent increases in cytotoxicity, chromosome damage and DNA double strand breaks in human lung cells. In response to zinc chromate-induced breaks, MRE11 expression was increased and ATM and ATR were phosphorylated, indicating that the DNA double strand break repair system was initiated in the cells. In addition, our data show that zinc chromate-induced double strand breaks were only observed in the G2/M phase population, with no significant amount of double strand breaks observed in G1 and S phase cells. These data will aid in understanding the mechanisms of zinc chromate toxicity and carcinogenesis.

  19. Synergistic effects of retinoic acid and tamoxifen on human breast cancer cells: Proteomic characterization

    SciTech Connect (OSTI)

    Wang Ying [Department of Chemistry, University of Hong Kong, Pokfulam, Hong Kong (China); He Qingyu [Department of Chemistry, University of Hong Kong, Pokfulam, Hong Kong (China); Chen Hongming [Department of Biochemistry, University of Vermont, College of Medicine, Burlington 05405 (United States); Chiu Jenfu [Department of Anatomy, The University of Hong Kong, Pokfulam, Hong Kong (China) and Department of Biochemistry, University of Vermont, College of Medicine, Burlington 05405 (United States)]. E-mail: jfchiu@hkucc.hku.hk

    2007-01-15

    The anti-estrogen tamoxifen and vitamin A-related compound, all-trans retinoic acid (RA), in combination act synergistically to inhibit the growth of MCF-7 human breast cancer cells. In the present study, we applied two-dimensional gel electrophoresis based proteomic approach to globally analyze this synergistic effect of RA and tamoxifen. Proteomic study revealed that multiple clusters of proteins were involved in RA and tamoxifen-induced apoptosis in MCF-7 breast cancer cells, including post-transcriptional and splicing factors, proteins related to cellular proliferation or differentiation, and proteins related to energy production and internal degradation systems. The negative growth factor-transforming growth factor {beta} (TGF{beta}) was secreted by RA and/or tamoxifen treatment and was studies as a potential mediator of the synergistic effects of RA and tamoxifen in apoptosis. By comparing protein alterations in treatments of RA and tamoxifen alone or in combination to those of TGF{beta} treatment, or co-treatment with TGF{beta} inhibitor SB 431542, proteomic results showed that a number of proteins were involved in TGF{beta} signaling pathway. These results provide valuable insights into the mechanisms of RA and tamoxifen-induced TGF{beta} signaling pathway in breast cancer cells.

  20. The critical roles of pre-replicative complex (pre-RC) component, Cdc6, in DNA replication and checkpoint response in human cells

    E-Print Network [OSTI]

    Lau, Eric Kirk

    2008-01-01

    In Molecular Pathology Eric Kirk Lau Committee in charge:in human cells by Eric Kirk Lau Doctor of Philosophy in

  1. Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2r??/? (NSG) Mice

    E-Print Network [OSTI]

    Drake, Adam

    Increasing demand for human hematopoietic stem cells (HSCs) in clinical and research applications necessitates expansion of HSCs in vitro. Before these cells can be used they must be carefully evaluated to assess their ...

  2. The number of cells in the human body is literally astronomical, about three orders of magnitude more than the number of stars in the Milky Way. Yet,

    E-Print Network [OSTI]

    Boal, David

    1 The number of cells in the human body is literally astronomical, about three orders of magnitude more than the number of stars in the Milky Way. Yet, for their immense number, the variety of cells is much smaller: only about 200 different cell types are represented in the collection of about 1014 cells

  3. The Dose-Response of Vitamin D on Cell Proliferation, Differentiation and Apoptosis in Human Osteosarcoma Cells

    E-Print Network [OSTI]

    Thompson, Lindsey M.

    2009-01-28

    ,25D in Osteosarcoma Cells ....................... 9 Mechanisms Involved in 1,25D Effects on Proliferation .......................... 11 Differentiation of Osteosarcoma Cells by 1,25D ..................................... 12 Apoptosis in Osteosarcoma... treatment with 1,25D or 25D in SaOS-2 and 143B cells ................................................................................ 38 FIGURE 12. Cell cycle distribution of SaOS-2 versus 143B control cells ................. 40 FIGURE 13. Working model...

  4. WNT5A inhibits human dental papilla cell proliferation and migration

    SciTech Connect (OSTI)

    Peng, L. [West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan (China) [West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan (China); State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, Sichuan (China); Ye, L.; Dong, G.; Ren, L.B.; Wang, C.L.; Xu, P. [West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan (China)] [West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan (China); Zhou, X.D., E-mail: pl_huaxi@yahoo.com.cn [West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan (China); State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, Sichuan (China)

    2009-12-18

    WNT proteins are a large family of cysteine-rich secreted molecules that are linked to both canonical and non-canonical signal pathways, and have been implicated in oncogenesis and tissue development. Canonical WNT proteins have been proven to play critical roles in tooth development, while little is known about the role of non-canonical WNT proteins such as WNT5A. In this study, WNT5A was localized to human dental papilla tissue and human dental papilla cells (HDPCs) cultured in vitro, using immunochemistry and RT-PCR. Recombinant adenovirus encoding full-length Wnt5a cDNA was constructed to investigate the biological role of WNT5A on HDPCs. The BrdU incorporation assay, the MTT assay and flow cytometric analysis showed that over-expression of Wnt5a strongly inhibited the proliferation of HDPCs in vitro. Wound healing and transwell migration assays indicated that over-expression of WNT5A reduced migration of HDPCs. In conclusion, our results showed that WNT5A negatively regulates both proliferation and migration of HDPCs, suggesting its important role in odontogenesis via controlling the HDPCs.

  5. Physics of the Structural Color on the Skin of Cephalopods 

    E-Print Network [OSTI]

    Gao, Meng 1981-

    2012-10-30

    by leucophore and iridophore cells, are important for cephalopod camou age; however, their scattering properties have not been very well studied. These colors are mainly due to the scattering of the speci c small scatterers inside of the cell. In this work we.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 1.2 Modeled cephalpod skin with the same sequence of cells as shown in the electron micrograph: chromatophore (a), iridophore (b), and leucophore (c)). The structural color is produced by iridophore (b) and leucophore (c...

  6. Human metastatic melanoma cell lines express high levels of growth hormone receptor and respond to GH treatment

    SciTech Connect (OSTI)

    Sustarsic, Elahu G.; Department of Biological Sciences, Ohio University, Athens, OH ; Junnila, Riia K.; Kopchick, John J.

    2013-11-08

    Highlights: •Most cancer types of the NCI60 have sub-sets of cell lines with high GHR expression. •GHR is highly expressed in melanoma cell lines. •GHR is elevated in advanced stage IV metastatic tumors vs. stage III. •GH treatment of metastatic melanoma cell lines alters growth and cell signaling. -- Abstract: Accumulating evidence implicates the growth hormone receptor (GHR) in carcinogenesis. While multiple studies show evidence for expression of growth hormone (GH) and GHR mRNA in human cancer tissue, there is a lack of quantification and only a few cancer types have been investigated. The National Cancer Institute’s NCI60 panel includes 60 cancer cell lines from nine types of human cancer: breast, CNS, colon, leukemia, melanoma, non-small cell lung, ovarian, prostate and renal. We utilized this panel to quantify expression of GHR, GH, prolactin receptor (PRLR) and prolactin (PRL) mRNA with real-time RT qPCR. Both GHR and PRLR show a broad range of expression within and among most cancer types. Strikingly, GHR expression is nearly 50-fold higher in melanoma than in the panel as a whole. Analysis of human metastatic melanoma biopsies confirmed GHR gene expression in melanoma tissue. In these human biopsies, the level of GHR mRNA is elevated in advanced stage IV tumor samples compared to stage III. Due to the novel finding of high GHR in melanoma, we examined the effect of GH treatment on three NCI60 melanoma lines (MDA-MB-435, UACC-62 and SK-MEL-5). GH increased proliferation in two out of three cell lines tested. Further analysis revealed GH-induced activation of STAT5 and mTOR in a cell line dependent manner. In conclusion, we have identified cell lines and cancer types that are ideal to study the role of GH and PRL in cancer, yet have been largely overlooked. Furthermore, we found that human metastatic melanoma tumors express GHR and cell lines possess active GHRs that can modulate multiple signaling pathways and alter cell proliferation. Based on this data, GH could be a new therapeutic target in melanoma.

  7. Formation of tRNA granules in the nucleus of heat-induced human cells

    SciTech Connect (OSTI)

    Miyagawa, Ryu [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan) [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Department of Biological Science, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8654 (Japan); Mizuno, Rie [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan)] [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Watanabe, Kazunori, E-mail: watanabe@ric.u-tokyo.ac.jp [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan)] [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Ijiri, Kenichi [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan) [Radioisotope Center, The University of Tokyo, 2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032 (Japan); Department of Biological Science, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8654 (Japan)

    2012-02-03

    Highlights: Black-Right-Pointing-Pointer tRNAs are tranlocated into the nucleus in heat-induced HeLa cells. Black-Right-Pointing-Pointer tRNAs form the unique granules in the nucleus. Black-Right-Pointing-Pointer tRNA ganules overlap with nuclear stress granules. -- Abstract: The stress response, which can trigger various physiological phenomena, is important for living organisms. For instance, a number of stress-induced granules such as P-body and stress granule have been identified. These granules are formed in the cytoplasm under stress conditions and are associated with translational inhibition and mRNA decay. In the nucleus, there is a focus named nuclear stress body (nSB) that distinguishes these structures from cytoplasmic stress granules. Many splicing factors and long non-coding RNA species localize in nSBs as a result of stress. Indeed, tRNAs respond to several kinds of stress such as heat, oxidation or starvation. Although nuclear accumulation of tRNAs occurs in starved Saccharomyces cerevisiae, this phenomenon is not found in mammalian cells. We observed that initiator tRNA{sup Met} (Meti) is actively translocated into the nucleus of human cells under heat stress. During this study, we identified unique granules of Meti that overlapped with nSBs. Similarly, elongator tRNA{sup Met} was translocated into the nucleus and formed granules during heat stress. Formation of tRNA granules is closely related to the translocation ratio. Then, all tRNAs may form the specific granules.

  8. Three Human Cell Types Respond to Multi-Walled Carbon Nanotubes and Titanium Dioxide Nanobelts with Cell-Specific Transcriptomic and Proteomic Expression Patterns.

    SciTech Connect (OSTI)

    Tilton, Susan C.; Karin, Norman J.; Tolic, Ana; Xie, Yumei; Lai, Xianyin; Hamilton, Raymond F.; Waters, Katrina M.; Holian, Andrij; Witzmann, Frank A.; Orr, Galya

    2014-08-01

    The growing use of engineered nanoparticles (NPs) in commercial and medical applications raises the urgent need for tools that can predict NP toxicity. Global transcriptome and proteome analyses were conducted on three human cell types, exposed to two high aspect ratio NP types, to identify patterns of expression that might indicate high versus low NP toxicity. Three cell types representing the most common routes of human exposure to NPs, including macrophage-like (THP-1), small airway epithelial and intestinal (Caco-2/HT29-MTX) cells, were exposed to TiO2 nanobelts (TiO2-NB; high toxicity) and multi-walled carbon nanotubes (MWCNT; low toxicity) at low (10 µg/mL) and high (100 µg/mL) concentrations for 1 and 24 h. Unique patterns of gene and protein expressions were identified for each cell type, with no differentially expressed (p < 0.05, 1.5-fold change) genes or proteins overlapping across all three cell types. While unique to each cell type, the early response was primarily independent of NP type, showing similar expression patterns in response to both TiO2-NB and MWCNT. The early response might, therefore, indicate a general response to insult. In contrast, the 24 h response was unique to each NP type. The most significantly (p < 0.05) enriched biological processes in THP-1 cells indicated TiO2-NB regulation of pathways associated with inflammation, apoptosis, cell cycle arrest, DNA replication stress and genomic instability, while MWCNT-regulated pathways indicated increased cell proliferation, DNA repair and anti-apoptosis. These two distinct sets of biological pathways might, therefore, underlie cellular responses to high and low NP toxicity, respectively.

  9. Capturing skin properties from dynamic mechanical analyses

    E-Print Network [OSTI]

    Sandford, Erika J. (Erika Jaye)

    2012-01-01

    Existing skin mechanical testing devices focus on measuring skin elasticity and are not tailored to assess the dynamic behavior of skin. The mathematical techniques used to analyze data collected using these devices are ...

  10. Comparative study of the chondrogenic potential of human bone marrow stromal cells, neonatal chondrocytes and adult chondrocytes

    SciTech Connect (OSTI)

    Saha, Sushmita; Kirkham, Jennifer; NIHR Leeds Musculoskeletal Biomedical Research Unit, University of Leeds, Chapel Allerton Hospital, Leeds LS74SA ; Wood, David; Curran, Stephen; Yang, Xuebin; NIHR Leeds Musculoskeletal Biomedical Research Unit, University of Leeds, Chapel Allerton Hospital, Leeds LS74SA

    2010-10-22

    Research highlights: {yields} This study has characterised three different cell types under conditions similar to those used for autologous chondrocyte implantation (ACI) for applications in cartilage repair/regeneration. {yields} Compared for the first time the chondrogenic potential of neonatal chondrocytes with human bone marrow stromal cells (HBMSCs) and adult chondrocytes. {yields} Demonstrated that adult chondrocytes hold greatest potential for use in ACI based on their higher proliferation rates, lower alkaline phosphatise activity and enhanced expression of chondrogenic genes. {yields} Demonstrated the need for chondroinduction as a necessary pre-requisite to efficient chondrogenesis in vitro and, by extrapolation, for cell based therapy (e.g. ACI or cartilage tissue engineering). -- Abstract: Cartilage tissue engineering is still a major clinical challenge with optimisation of a suitable source of cells for cartilage repair/regeneration not yet fully addressed. The aims of this study were to compare and contrast the differences in chondrogenic behaviour between human bone marrow stromal cells (HBMSCs), human neonatal and adult chondrocytes to further our understanding of chondroinduction relative to cell maturity and to identify factors that promote chondrogenesis and maintain functional homoeostasis. Cells were cultured in monolayer in either chondrogenic or basal medium, recapitulating procedures used in existing clinical procedures for cell-based therapies. Cell doubling time, morphology and alkaline phosphatase specific activity (ALPSA) were determined at different time points. Expression of chondrogenic markers (SOX9, ACAN and COL2A1) was compared via real time polymerase chain reaction. Amongst the three cell types studied, HBMSCs had the highest ALPSA in basal culture and lowest ALPSA in chondrogenic media. Neonatal chondrocytes were the most proliferative and adult chondrocytes had the lowest ALPSA in basal media. Gene expression analysis revealed a difference in the temporal expression of chondrogenic markers which were up regulated in chondrogenic medium compared to levels in basal medium. Of the three cell types studied, adult chondrocytes offer a more promising cell source for cartilage tissue engineering. This comparative study revealed differences between the microenvironment of all three cell types and provides useful information to inform cell-based therapies for cartilage regeneration.

  11. Three-Dimensional Model on Thermal Response of Skin Subject to Laser Heating

    E-Print Network [OSTI]

    Zhang, Jun

    thermal response. The time-dependent equation is discretized using the #12;nite di#11;erence methodThree-Dimensional Model on Thermal Response of Skin Subject to Laser Heating #3; Wensheng Shen y to investigate the transient thermal response of human skin subject to laser heating. The temperature

  12. Group B Streptococcal b-Hemolysin/Cytolysin Promotes Invasion of Human Lung Epithelial Cells and the Release of Interleukin-8

    E-Print Network [OSTI]

    Nizet, Victor

    Group B Streptococcal b-Hemolysin/Cytolysin Promotes Invasion of Human Lung Epithelial Cells and lung injury are hallmarks of early-onset neonatal group B streptococcal (GBS) infections. Production. To elucidate the contribution of the b-h/c toxin to lung injury, the interactions of GBS wild-type strains

  13. Cell Biophysics for Label-free Single-Cell Analysis and Sorting

    E-Print Network [OSTI]

    Masaeli, Mahdokht

    2013-01-01

    activated T cells in psoriasis vulgaris lesional skin: bloodin diseases such as psoriasis and lupus using multicolor

  14. PPAR{gamma} ligands induce growth inhibition and apoptosis through p63 and p73 in human ovarian cancer cells

    SciTech Connect (OSTI)

    Kim, Soyeon; Innovative Research Institute for Cell Therapy, Seoul National University College of Medicine and Hospital, Seoul ; Lee, Jae-Jung; Heo, Dae Seog

    2011-03-18

    Research highlights: {yields} PPAR{gamma} ligands increased the rate of apoptosis and inhibition of proliferation in ovarian cancer cells. {yields} PPAR{gamma} ligands induced p63 and p73 expression, but not p53. {yields} p63 and p73 leads to an increase in p21 expression and apoptosis in ovarian cancer cells with treatment PPAR{gamma} ligands. {yields} These findings suggest that PPAR{gamma} ligands suppressed growth of ovarian cancer cells through upregulation of p63 and p73. -- Abstract: Peroxisome proliferator-activated receptor gamma (PPAR{gamma}) agonists, including thiazolidinediones (TZDs), can induce anti-proliferation, differentiation, and apoptosis in various cancer cell types. This study investigated the mechanism of the anticancer effect of TZDs on human ovarian cancer. Six human ovarian cancer cell lines (NIH:OVCAR3, SKOV3, SNU-251, SNU-8, SNU-840, and 2774) were treated with the TZD, which induced dose-dependent inhibition of cell growth. Additionally, these cell lines exhibited various expression levels of PPAR{gamma} protein as revealed by Western blotting. Flow cytometry showed that the cell cycle was arrested at the G1 phase, as demonstrated by the appearance of a sub-G1 peak. This observation was corroborated by the finding of increased levels of Bax, p21, PARP, and cleaved caspase 3 in TGZ-treated cells. Interestingly, when we determined the effect of p53-induced growth inhibition in these three human ovarian cancer cells, we found that they either lacked p53 or contained a mutant form of p53. Furthermore, TGZ induced the expression of endogenous or exogenous p63 and p73 proteins and p63- or p73-directed short hairpin (si) RNAs inhibited the ability of TGZ to regulate expression of p21 in these cells. Thus, our results suggest that PPAR{gamma} ligands can induce growth suppression of ovarian cancer cells and mediate p63 and p73 expression, leading to enhanced growth inhibition and apoptosis. The tumor suppressive effects of PPAR{gamma} ligands may have applications for the treatment of ovarian cancer.

  15. MiR-18a regulates the proliferation, migration and invasion of human glioblastoma cell by targeting neogenin

    SciTech Connect (OSTI)

    Song, Yichen; Wang, Ping; Zhao, Wei; Yao, Yilong; Liu, Xiaobai; Ma, Jun; Xue, Yixue; Liu, Yunhui

    2014-05-15

    MiR-17-92 cluster has recently been reported as an oncogene in some tumors. However, the association of miR-18a, an important member of this cluster, with glioblastoma remains unknown. Therefore, this study aims to investigate the expression of miR-18a in glioblastoma and its role in biological behavior of U87 and U251 human glioblastoma cell lines. Quantitative RT-PCR results showed that miR-18a was highly expressed in glioblastoma tissues and U87 and U251 cell lines compared with that in human brain tissues and primary normal human astrocytes, and the expression levels were increased along with the rising pathological grades of glioblastoma. Neogenin was identified as the target gene of miR-18a by dual-luciferase reporter assays. RT-PCR and western blot results showed that its expression levels were decreased along with the rising pathological grades of glioblastoma. Inhibition of miR-18a expression was established by transfecting exogenous miR-18a inhibitor into U87 and U251 cells, and its effects on the biological behavior of glioblastoma cells were studied using CCK-8 assay, transwell assay and flow cytometry. Inhibition of miR-18a expression in U87 and U251 cells significantly up-regulated neogenin, and dramatically suppressed the abilities of cell proliferation, migration and invasion, induced cell cycle arrest and promoted cellular apoptosis. Collectively, these results suggest that miR-18a may regulate biological behavior of human glioblastoma cells by targeting neogenin, and miR-18a can serve as a potential target in the treatment of glioblastoma. - Highlights: • MiR-18a was highly expressed in glioblastoma tissues and U87 and U251 cell lines. • Neogenin was identified as the target gene of miR-18a. • Neogenin expressions were decreased along with the rising pathological grades of glioblastoma. • Inhibition of miR-18a suppressed biological behavior of glioma cells by up-regulating neogenin.

  16. MicroRNA-320a suppresses human colon cancer cell proliferation by directly targeting {beta}-catenin

    SciTech Connect (OSTI)

    Sun, Jian-Yong; State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, 710032 Xi'an ; Huang, Yi; Li, Ji-Peng; Zhang, Xiang; Wang, Lei; Meng, Yan-Ling; Yan, Bo; Bian, Yong-Qian; Zhao, Jing; Wang, Wei-Zhong; and others

    2012-04-20

    Highlights: Black-Right-Pointing-Pointer miR-320a is downregulated in human colorectal carcinoma. Black-Right-Pointing-Pointer Overexpression of miR-320a inhibits colon cancer cell proliferation. Black-Right-Pointing-Pointer {beta}-Catenin is a direct target of miR-320a in colon cancer cells. Black-Right-Pointing-Pointer miR-320a expression inversely correlates with mRNA expression of {beta}-catenin's target genes in human colon carcinoma. -- Abstract: Recent profile studies of microRNA (miRNA) expression have documented a deregulation of miRNA (miR-320a) in human colorectal carcinoma. However, its expression pattern and underlying mechanisms in the development and progression of colorectal carcinoma has not been elucidated clearly. Here, we performed real-time PCR to examine the expression levels of miR-320a in colon cancer cell lines and tumor tissues. And then, we investigated its biological functions in colon cancer cells by a gain of functional strategy. Further more, by the combinational approaches of bioinformatics and experimental validation, we confirmed target associations of miR-320a in colorectal carcinoma. Our results showed that miR-320a was frequently downregulated in cancer cell lines and colon cancer tissues. And we demonstrated that miR-320a restoration inhibited colon cancer cell proliferation and {beta}-catenin, a functionally oncogenic molecule was a direct target gene of miR-320a. Finally, the data of real-time PCR showed the reciprocal relationship between miR-320a and {beta}-catenin's downstream genes in colon cancer tissues. These findings indicate that miR-320a suppresses the growth of colon cancer cells by directly targeting {beta}-catenin, suggesting its application in prognosis prediction and cancer treatment.

  17. S.N.A.K.E. : a dynamically reconfigurable Artificial Sensate Skin

    E-Print Network [OSTI]

    Barroeta Pérez, Gerardo

    2006-01-01

    The idea of an Artificial Sensate Skin device that mimics the characteristics and functions of its analogous living tissue whether human or animal is not new. Yet, most of the current related work has been focused in the ...

  18. Sensitive skins and somatic processing for affective and sociable robots based upon a somatic alphabet approach

    E-Print Network [OSTI]

    Stiehl, Walter Daniel, 1980-

    2005-01-01

    The sense of touch is one of the most important senses of the human body. This thesis describes the biologically inspired design of "sensitive skins" for two different robotic platforms: Leonardo, a high degree-of-freedom, ...

  19. Study the cytotoxicity of different kinds of water-soluble nanoparticles in human osteoblast-like MG-63 cells

    SciTech Connect (OSTI)

    Niu, Lu; Li, Yang; Li, Xiaojie; Gao, Xue; Su, Xingguang

    2012-11-15

    Highlights: ? Preparation of three kinds of water-soluble QDs: CdTe, CdTe@SiO{sub 2}, Mn:ZnSe. ? Evaluated the cytotoxicity qualitatively and quantitatively. ? Fluorescent staining. ? Detected the total intracellular cadmium in cells. -- Abstract: Quantum nanoparticles have been applied extensively in biological and medical fields, the cytotoxicity of nanoparticles becomes the key point we should concern. In this paper, the cytotoxicity of three kinds of water-soluble nanoparticles: CdTe, CdTe@SiO{sub 2} and Mn:ZnSe was studied. We evaluated the nanoparticles toxicity qualitatively by observing the morphological changes of human osteoblast-like MG-63 cells at different incubation times and colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays were carried out to detect the cell viability quantitatively. The results showed that CdTe nanoparticles with high concentrations caused cells to die largely while CdTe@SiO{sub 2} and Mn:ZnSe nanoparticles had no obvious effect. For further study, we studied the relation between the cell viability and the total cadmium concentration in cells and found that the viability of cells treated with CdTe@SiO{sub 2} nanoparticles was higher than that treated with CdTe nanoparticles. We also discovered that the death rate of cells co-incubated with CdTe nanoparticles was proportional to the total intracellular cadmium concentrations.

  20. Lysophospholipid presentation by CD1d and recognition by a human Natural Killer T-cell receptor

    SciTech Connect (OSTI)

    López-Sagaseta, Jacinto; Sibener, Leah V.; Kung, Jennifer E.; Gumperz, Jenny; Adams, Erin J. (UC); (UW-MED)

    2014-10-02

    Invariant Natural Killer T (iNKT) cells use highly restricted {alpha}{beta} T cell receptors (TCRs) to probe the repertoire of lipids presented by CD1d molecules. Here, we describe our studies of lysophosphatidylcholine (LPC) presentation by human CD1d and its recognition by a native, LPC-specific iNKT TCR. Human CD1d presenting LPC adopts an altered conformation from that of CD1d presenting glycolipid antigens, with a shifted {alpha}1 helix resulting in an open A pocket. Binding of the iNKT TCR requires a 7-{angstrom} displacement of the LPC headgroup but stabilizes the CD1d-LPC complex in a closed conformation. The iNKT TCR CDR loop footprint on CD1d-LPC is anchored by the conserved positioning of the CDR3{alpha} loop, whereas the remaining CDR loops are shifted, due in part to amino-acid differences in the CDR3{beta} and J{beta} segment used by this iNKT TCR. These findings provide insight into how lysophospholipids are presented by human CD1d molecules and how this complex is recognized by some, but not all, human iNKT cells.

  1. A Computational Model Incorporating Neural Stem Cell Dynamics Reproduces Glioma Incidence across the Lifespan in the Human Population

    E-Print Network [OSTI]

    Bauer, Roman; Stoll, Elizabeth

    2015-01-01

    Glioma is the most common form of primary brain tumor. Demographically, the risk of occurrence increases until old age. Here we present a novel computational model to reproduce the probability of glioma incidence across the lifespan. Previous mathematical models explaining glioma incidence are framed in a rather abstract way, and do not directly relate to empirical findings. To decrease this gap between theory and experimental observations, we incorporate recent data on cellular and molecular factors underlying gliomagenesis. Since evidence implicates the adult neural stem cell as the likely cell-of-origin of glioma, we have incorporated empirically-determined estimates of neural stem cell number, cell division rate, mutation rate and oncogenic potential into our model. We demonstrate that our model yields results which match actual demographic data in the human population. In particular, this model accounts for the observed peak incidence of glioma at approximately 80 years of age, without the need to assert...

  2. Multipotent human stromal cells improve cardiac function after myocardial infarction in mice without long-term engraftment

    SciTech Connect (OSTI)

    Iso, Yoshitaka [Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States); Cardiovascular Research Institute, Department of Medicine, University of Vermont, 208 South Park Drive, Colchester, VT 05446 (United States); Spees, Jeffrey L. [Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States); Cardiovascular Research Institute, Department of Medicine, University of Vermont, 208 South Park Drive, Colchester, VT 05446 (United States); E-mail: Jeffrey.Spees@uvm.edu; Serrano, Claudia [Section of Cardiology, Department of Medicine, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States); Bakondi, Benjamin [Cardiovascular Research Institute, Department of Medicine, University of Vermont, 208 South Park Drive, Colchester, VT 05446 (United States); Pochampally, Radhika [Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States); Song, Yao-Hua [Section of Cardiology, Department of Medicine, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States); Sobel, Burton E. [Cardiovascular Research Institute, Department of Medicine, University of Vermont, 208 South Park Drive, Colchester, VT 05446 (United States); Delafontaine, Patrick [Section of Cardiology, Department of Medicine, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States); Prockop, Darwin J. [Center for Gene Therapy, Tulane University Health Sciences Center, 1430 Tulane Avenue, New Orleans, LA 70112 (United States)]. E-mail: dprocko@tulane.edu

    2007-03-16

    The aim of this study was to determine whether intravenously administered multipotent stromal cells from human bone marrow (hMSCs) can improve cardiac function after myocardial infarction (MI) without long-term engraftment and therefore whether transitory paracrine effects or secreted factors are responsible for the benefit conferred. hMSCs were injected systemically into immunodeficient mice with acute MI. Cardiac function and fibrosis after MI in the hMSC-treated group were significantly improved compared with controls. However, despite the cardiac improvement, there was no evident hMSC engraftment in the heart 3 weeks after MI. Microarray assays and ELISAs demonstrated that multiple protective factors were expressed and secreted from the hMSCs in culture. Factors secreted by hMSCs prevented cell death of cultured cardiomyocytes and endothelial cells under conditions that mimicked tissue ischemia. The favorable effects of hMSCs appear to reflect the impact of secreted factors rather than engraftment, differentiation, or cell fusion.

  3. Three-dimensional Invasion of Human Glioblastoma Cells Remains Unchanged by X-ray and Carbon Ion Irradiation In Vitro

    SciTech Connect (OSTI)

    Eke, Iris; Storch, Katja; Kaestner, Ina; Vehlow, Anne; Faethe, Christina; Mueller-Klieser, Wolfgang; Taucher-Scholz, Gisela; Temme, Achim; Schackert, Gabriele; Department of Radiation Oncology, Medical Faculty Carl Gustav Carus, Dresden University of Technology, Dresden

    2012-11-15

    Purpose: Cell invasion represents one of the major determinants that treatment has failed for patients suffering from glioblastoma. Contrary findings have been reported for cell migration upon exposure to ionizing radiation. Here, the migration and invasion capability of glioblastoma cells on and in collagen type I were evaluated upon irradiation with X-rays or carbon ions. Methods and Materials: Migration on and invasion in collagen type I were evaluated in four established human glioblastoma cell lines exposed to either X-rays or carbon ions. Furthermore, clonogenic radiation survival, proliferation (5-bromo-2-deoxyuridine positivity), DNA double-strand breaks ({gamma}H2AX/53BP1-positive foci), and expression of invasion-relevant proteins (eg, {beta}1 integrin, FAK, MMP2, and MMP9) were explored. Migration and invasion assays for primary glioblastoma cells also were carried out with X-ray irradiation. Results: Neither X-ray nor carbon ion irradiation affected glioblastoma cell migration and invasion, a finding similarly observed in primary glioblastoma cells. Intriguingly, irradiated cells migrated unhampered, despite DNA double-strand breaks and reduced proliferation. Clonogenic radiation survival was increased when cells had contact with extracellular matrix. Specific inhibition of the {beta}1 integrin or proliferation-associated signaling molecules revealed a critical function of JNK, PI3K, and p38 MAPK in glioblastoma cell invasion. Conclusions: These findings indicate that X-rays and carbon ion irradiation effectively reduce proliferation and clonogenic survival without modifying the migration and invasion ability of glioblastoma cells in a collagen type I environment. Addition of targeted agents against members of the MAPK and PI3K signaling axis to conventional chemoradiation therapy seems potentially useful to optimize glioblastoma therapy.

  4. Skinning Arbitrary Deformations Ladislav Kavan 1,2 Rachel McDonnell1 Simon Dobbyn1 Jiri Zara2 Carol O'Sullivan1

    E-Print Network [OSTI]

    Plotkin, Joshua B.

    , such as moving human or animal figures. In this paper, we demonstrate how to automatically construct skinning way of representing the animation of quasi- articulated objects, such as virtual characters, is known as skinning (or matrix palette skinning). It is based on the observation that an animation of a virtual

  5. Methylation patterns and mathematical models reveal dynamics of stem cell turnover in the human colon

    E-Print Network [OSTI]

    renewal while the daughter stem cell remains in the niche; each stem cell is ``immortal'' under this modelCommentary Methylation patterns and mathematical models reveal dynamics of stem cell turnover in diseases such as cancer, it is essential to understand the process of somatic cell development and renewal

  6. Up-regulation of E-cadherin by small activating RNA inhibits cell invasion and migration in 5637 human bladder cancer cells

    SciTech Connect (OSTI)

    Mao Qiqi; Li Yubing; Zheng Xiangyi; Yang Kai; Shen Huafeng; Qin Jie; Bai Yu; Kong Debo; Jia Xiaolong [Department of Urology, First Affiliated Hospital, School of Medicine, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China); Xie Liping [Department of Urology, First Affiliated Hospital, School of Medicine, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China)], E-mail: xielp@zjuem.zju.edu.cn

    2008-10-31

    Recent studies have reported that chemically synthesized small duplex RNAs complementary to promoters of target genes can specifically induce gene expression in several cancer cell lines. Such dsRNA, referred to as small activating RNA (saRNA), are involved in the recently described phenomenon called RNA activation (RNAa). Recent findings show that saRNA can inhibit cell proliferation and viability via up-regulation of p21{sup WAF1/CIP1} (p21) in human bladder cancer cells. In the present study, we demonstrate that induction of E-cadherin expression by saRNA leads to suppression of migration and invasion of 5637 human bladder cancer cells in vitro. The elevated E-cadherin expression was confirmed at transcriptional and protein levels after transfection of a 21-nucleotide dsRNA targeting the E-cadherin promoter (dsEcad). Furthermore, this inhibitory effect was associated with relocalization of {beta}-catenin from the nucleus to the plasma membrane and decreased {beta}-catenin-mediated transactivation. These data suggest that activation of E-cadherin by saRNA may have a therapeutic benefit for bladder and other types of cancer.

  7. Human epididymis protein 4 (HE4) plays a key role in ovarian cancer cell adhesion and motility

    SciTech Connect (OSTI)

    Lu, Renquan; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032 ; Sun, Xinghui; Department of Medicine, Harvard Medical School, MA 02115 ; Xiao, Ran; Zhou, Lei; Gao, Xiang; Guo, Lin; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032

    2012-03-09

    Highlights: Black-Right-Pointing-Pointer We generated stable transduced HE4 overexpression and knockdown cells. Black-Right-Pointing-Pointer HE4 was associated with EOC cell adhesion and motility. Black-Right-Pointing-Pointer HE4 might have some effects on activation of EGFR-MAPK signaling pathway. Black-Right-Pointing-Pointer HE4 play an important role in EOC tumorigenicity. -- Abstract: Human epididymis protein 4 (HE4) is a novel and specific biomarker for epithelial ovarian cancer (EOC). We previously demonstrated that serum HE4 levels were significantly elevated in the majority of EOC patients but not in subjects with benign disease or healthy controls. However, the precise mechanism of HE4 protein function is unknown. In this study, we generated HE4-overexpressing SKOV3 cells and found that stably transduced cells promoted cell adhesion and migration. Knockdown of HE4 expression was achieved by stable transfection of SKOV3 cells with a construct encoding a short hairpin DNA directed against the HE4 gene. Correspondingly, the proliferation and spreading ability of HE4-expressed cells were inhibited by HE4 suppression. Mechanistically, impaired EGFR and Erk1/2 phosphorylation were observed in cells with HE4 knockdown. The phosphorylation was restored when the knockdown cells were cultured in conditioned medium containing HE4. Moreover, in vivo tumorigenicity showed that HE4 suppression markedly inhibited the growth of tumors. This suggests that expression of HE4 is associated with cancer cell adhesion, migration and tumor growth, which can be related to its effects on the EGFR-MAPK signaling pathway. Our results provide evidence of the cellular and molecular mechanisms that may underlie the motility-promoting role of HE4 in EOC progression. The role of HE4 as a target for gene-based therapy might be considered in future studies.

  8. Sensitive Targeted Quantification of ERK Phosphorylation Dynamics and Stoichiometry in Human Cells without Affinity Enrichment

    SciTech Connect (OSTI)

    Shi, Tujin; Gao, Yuqian; Gaffrey, Matthew J.; Nicora, Carrie D.; Fillmore, Thomas L.; Chrisler, William B.; Gritsenko, Marina A.; Wu, Chaochao; He, Jintang; Bloodsworth, Kent J.; Zhao, Rui; Camp, David G.; Liu, Tao; Rodland, Karin D.; Smith, Richard D.; Wiley, H. S.; Qian, Weijun

    2014-12-17

    Mass spectrometry-based targeted quantification is a promising technology for site-specific quantification of posttranslational modifications (PTMs). However, a major constraint of most targeted MS approaches is the limited sensitivity for quantifying low-abundance PTMs, requiring the use of affinity reagents to enrich specific PTMs. Herein, we demonstrate the direct site-specific quantification of ERK phosphorylation isoforms (pT, pY, pTpY) and their relative stoichiometries using a highly sensitive targeted MS approach termed high-pressure, high-resolution separations with intelligent selection and multiplexing (PRISM). PRISM provides effective enrichment of target peptides within a given fraction from complex biological matrix with minimal sample losses, followed by selected reaction monitoring (SRM) quantification. The PRISM-SRM approach enabled direct quantification of ERK phosphorylation in human mammary epithelial cells (HMEC) from as little as 25 µg tryptic peptides from whole cell lysates. Compared to immobilized metal-ion affinity chromatography, PRISM provided >10-fold improvement in signal intensities, presumably due to the better peptide recovery of PRISM for handling small size samples. This approach was applied to quantify ERK phosphorylation dynamics in HMEC treated by different doses of EGF at both the peak activation (10 min) and steady state (2 h). At 10 min, the maximal ERK activation was observed with 0.3 ng/mL dose, whereas the maximal steady state level of ERK activation at 2 h was at 3 ng/ml dose, corresponding to 1200 and 9000 occupied receptors, respectively. At 10 min, the maximally activated pTpY isoform represented ~40% of total ERK, falling to less than 10% at 2 h. The time course and dose-response profiles of individual phosphorylated ERK isoforms indicated that singly phosphorylated pT-ERK never increases significantly, while the increase of pY-ERK paralleled that of pTpY-ERK. This data supports for a processive, rather than distributed, model of ERK phosphorylation. The PRISM-SRM quantification of protein phosphorylation illustrates the potential for simultaneous quantification of multiple PTMs.

  9. AUTOMATED MONITORING OF HUMAN EMBRYONIC CELLS UP TO THE 5-CELL STAGE IN TIME-LAPSE MICROSCOPY IMAGES

    E-Print Network [OSTI]

    Gould, Stephen

    fertilization (IVF) treatments. Automated prediction of the embryo viability, by tracking cell divisions up infertility. The suc- cess of these methods relies on identifying the most viable embryos. In most cases

  10. Chronic exposure to bisphenol A reduces SULT1A1 activity in the human placental cell line BeWo

    E-Print Network [OSTI]

    Mitra, Pallabi

    2006-10-27

    Chronic exposure to bisphenol A reduces SULT1A1 activity in the human placental cell line BeWo Pallabi Mitra Department of Pharmaceutical Chemistry University of Kansas October 27, 2006 Outline ? Placental structure and models ? Placental... permeation ? Placental metabolism and regulation (induction/inhibition) ? Sulfotransferase enzymes in trophoblast ? Bisphenol A ? Effects of bisphenol A on SULT1A1 ? Conclusions The placental barrier Mother?s blood ?Trophoblasts and syncytiotrophoblasts line...

  11. Human T follicular helper and T follicular regulatory cell maintenance is independent of germinal centres

    E-Print Network [OSTI]

    Wallin, Elizabeth F.; Jolly, Elaine C.; Suchánek, Ond?ej; Bradley, J. Andrew; Espéli, Marion; Jayne, David R. W.; Linterman, Michelle A.; Smith, Kenneth G. C.

    2014-09-15

    and the number of Tfh cells45, because Tfh maintenance requires sustained antigenic stimulation from GC B cells22. We were therefore interested in whether the lack of GC B cells in rituximab treated patients reduced GC Tfh numbers. Although CXCR5 and ICOS... of the absence of GC B cells and the reports that rituximab can distort follicular architecture49, we have used CD57 as a marker of GC-Tfh cells, in combination with the Tfh/Tfr surface receptor CXCR5. Although the function of CD57 on Tfh cells is not well...

  12. Plant cyclopeptide RA-V kills human breast cancer cells by inducing mitochondria-mediated apoptosis through blocking PDK1–AKT interaction

    SciTech Connect (OSTI)

    Fang, Xian-Ying; Chen, Wei; Fan, Jun-Ting; Song, Ran; Wang, Lu; Gu, Yan-Hong; Zeng, Guang-Zhi; Shen, Yan; Wu, Xue-Feng; Tan, Ning-Hua; Xu, Qiang; Sun, Yang

    2013-02-15

    In the present paper, we examined the effects of a natural cyclopeptide RA-V on human breast cancer cells and the underlying mechanisms. RA-V significantly inhibited the growth of human breast cancer MCF-7, MDA-MB-231 cells and murine breast cancer 4T1 cells. In addition, RA-V triggered mitochondrial apoptotic pathway which was indicated by the loss of mitochondrial membrane potential, the release of cytochrome c, and the activation of caspase cascade. Further study showed that RA-V dramatically inhibited phosphorylation of AKT and 3-phosphoinositide dependent protein kinase 1 (PDK1) in MCF-7 cells. Moreover, RA-V disrupted the interaction between PDK1 and AKT in MCF-7 cells. Furthermore, RA-V-induced apoptosis could be enhanced by phosphatidylinositol 3-kinase inhibitor or attenuated by over-expression of AKT in all the three kinds of breast cancer cells. Taken together, this study shows that RA-V, which can induce mitochondria-mediated apoptosis, exerts strong anti-tumor activity against human breast cancer. The underlying anti-cancer mechanism of RA-V is related to the blockage of the interaction between PDK1 and AKT. - Highlights: ? Plant cyclopeptide RA-V kills human breast cancer cells. ? RA-V triggered mitochondrial apoptotic pathway in human breast cancer cells. ? RA-V inhibited phosphorylation of AKT and PDK1 in breast cancer MCF-7 cells. ? Its mechanism is related to the blockage of the interaction between PDK1 and AKT.

  13. A critical comparison of human face rendering techniques

    E-Print Network [OSTI]

    Arizpe, Arturo Andrew

    2006-01-01

    Human skin exhibits complex light reflectance properties that make it difficult to render realistically. In recent years, many techniques have been introduced to render skin, with varying degrees of complexity and realism. ...

  14. Induction of apoptosis by esculetin in human leukemia U937 cells through activation of JNK and ERK

    SciTech Connect (OSTI)

    Park, Cheol; Jin, Cheng-Yun [Division of Biological Sciences, College of Natural Science, Pusan National University, Busan 609-735 (Korea, Republic of); Kim, Gi-Young [Faculty of Applied Marine Science, Cheju National University, Jeju Special Self-Governing Province 690-756 (Korea, Republic of); Choi, Il-Whan [Department of Microbiology, College of Medicine and Center for Viral Disease Research, Inje University, Busan 614-735 (Korea, Republic of); Kwon, Taeg Kyu [Department of Immunology, School of Medicine, Keimyung University, Taegu 700-712 (Korea, Republic of); Choi, Byung Tae [Department of Anatomy, Graduate School of Oriental Medicine, Pusan National University, 609-735 (Korea, Republic of); Lee, Su Jae [Department of Chemistry, Hanyang University, Seoul 133-791 (Korea, Republic of); Lee, Won Ho [Division of Biological Sciences, College of Natural Science, Pusan National University, Busan 609-735 (Korea, Republic of); Choi, Yung Hyun [Department of Anatomy, Graduate School of Oriental Medicine, Pusan National University, 609-735 (Korea, Republic of); Department of Biochemistry, Dongeui University College of Oriental Medicine and Department of Biomaterial Control (BK21 program), Dongeui University Graduate School, Busan 614-052 (Korea, Republic of)], E-mail: lab301@nate.com

    2008-03-01

    Esculetin is a phenolic compound that is found in various natural plant products and induces apoptosis in several types of human cancer cells. However, the underlying mechanisms of its action are not completely understood. In the present study, we used human leukemia cells to gain further insight into the mechanism of esculetin-induced anti-proliferative action and apoptosis. It was found that esculetin inhibits cell viability by inducing apoptosis, as evidenced by the formation of apoptotic bodies, DNA fragmentation, and the accumulation of cells in the sub-G1 phase. Esculetin-induced apoptosis was correlated with mitochondrial dysfunction, leading to the release of cytochrome c from the mitochondria to the cytosol, as well as the proteolytic activation of caspases. The z-DEVD-fmk caspase-3 inhibitor and the ectopic expression of anti-apoptotic Bcl-2 significantly inhibited esculetin-induced apoptosis, demonstrating the important role of caspase-3 and mitochondrial proteins in the observed cytotoxic effect. Furthermore, esculetin selectively increased the phosphorylation of extracellular-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK), but not that of other kinases such as Akt and p38 activation. In addition, an ERK-specific inhibitor, PD98059, and a JNK-specific inhibitor, SP600125, showed inhibited sub-G1 phase DNA content, DNA fragmentation, caspase activation, and mitochondrial dysfunction induced by esculetin treatment. These results indicated that the JNK and ERK pathways were key regulators of apoptosis in response to esculetin in human leukemia U937 cells.

  15. Modulation of dendritic cells by human neutrophil elastase and its inhibitors in pulmonary inflammation 

    E-Print Network [OSTI]

    Roghanian, Ali

    2007-01-01

    Dendritic cells (DC) are sentinels of the immune system that display an extraordinary capacity to present antigen to naïve T cells and initiate immune responses. DCs are distributed throughout the lungs in the conducting ...

  16. LET dependence of radiation-induced bystander effects using human prostate tumor cells

    E-Print Network [OSTI]

    Anzenberg, Vered

    2008-01-01

    In the past fifteen years, evidence provided by many independent research groups have indicated higher numbers of cells exhibiting damage than expected based on the number of cells traversed by the radiation. This phenomenon ...

  17. Differential modulation of mediator release from human basophils and mast cells by mizolastine

    E-Print Network [OSTI]

    Gelb, Michael

    release from basophils and mast cells. Objective To study the in vitro effects of mizolastine, an H1 and bronchoconstrictor agents and they modulate local immune responses and inflammatory cell infiltration [1 cells [16] and reduce eosinophil infiltration during the late-phase response [17]. These data indicate that a

  18. Dissecting Biological Dark Matter: Single Cell Genetic Analysis of TM7, a Rare and Uncultivated Microbe from the Human Mouth

    SciTech Connect (OSTI)

    Fenner, Marsha W; Marcy, Yann; Ouverney, Cleber; Bik, Elisabeth M.; Losekann, Tina; Ivanova, Natalia; Martin, H. Garcia; Szeto, E.; Platt, Darren; Hugenholtz, Philip; Relman, David A.; Quake, Stephen R.

    2007-07-01

    We have developed a microfluidic device that allows the isolation and genome amplification of individual microbial cells, thereby enabling organism-level genomic analysis of complex microbial ecosystems without the need for culture. This device was used to perform a directed survey of the human subgingival crevice and to isolate bacteria having rod-like morphology. Several isolated microbes had a 16S rRNA sequence that placed them in candidate phylum TM7, which has no cultivated or sequenced members. Genome amplification from individual TM7 cells allowed us to sequence and assemble >1,000 genes, providing insight into the physiology of members of this phylum. This approach enables single-cell genetic analysis of any uncultivated minority member of a microbial community.

  19. Characterization of cancer stem cell properties of CD24 and CD26-positive human malignant mesothelioma cells

    SciTech Connect (OSTI)

    Yamazaki, Hiroto; Naito, Motohiko; Ghani, Farhana Ishrat [Division of Clinical Immunology, Institute of Medical Science, University of Tokyo, Tokyo (Japan)] [Division of Clinical Immunology, Institute of Medical Science, University of Tokyo, Tokyo (Japan); Dang, Nam H. [Division of Hematology/Oncology, University of Florida Shands Cancer Center, Gainesville, FL 32610 (United States)] [Division of Hematology/Oncology, University of Florida Shands Cancer Center, Gainesville, FL 32610 (United States); Iwata, Satoshi [Division of Clinical Immunology, Institute of Medical Science, University of Tokyo, Tokyo (Japan)] [Division of Clinical Immunology, Institute of Medical Science, University of Tokyo, Tokyo (Japan); Morimoto, Chikao, E-mail: morimoto@ims.u-tokyo.ac.jp [Division of Clinical Immunology, Institute of Medical Science, University of Tokyo, Tokyo (Japan)] [Division of Clinical Immunology, Institute of Medical Science, University of Tokyo, Tokyo (Japan)

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer We focused on CD24 and CD26 for further analysis of CSC properties in MM. Black-Right-Pointing-Pointer Their expressions were correlated with chemoresistance, cell growth, and invasion. Black-Right-Pointing-Pointer Their expressions were also correlated with several cancer related genes. Black-Right-Pointing-Pointer The expression of each marker was correlated with different CSC property in Meso1. Black-Right-Pointing-Pointer Phosphorylation of ERK by EGF was regulated by expression of CD26, but not CD24. -- Abstract: Malignant mesothelioma (MM) is an asbestos-related malignancy characterized by rapid growth and poor prognosis. In our previous study, we have demonstrated that several cancer stem cell (CSC) markers correlated with CSC properties in MM cells. Among these markers, we focused on two: CD24, the common CSC marker, and CD26, the additional CSC marker. We further analyzed the CSC properties of CD24 and CD26-positve MM cells. We established RNAi-knockdown cells and found that these markers were significantly correlated with chemoresistance, proliferation, and invasion potentials in vitro. Interestingly, while Meso-1 cells expressed both CD24 and CD26, the presence of each of these two markers was correlated with different CSC property. In addition, downstream signaling of these markers was explored by microarray analysis, which revealed that their expressions were correlated with several cancer-related genes. Furthermore, phosphorylation of ERK by EGF stimulation was significantly affected by the expression of CD26, but not CD24. These results suggest that CD24 and CD26 differentially regulate the CSC potentials of MM and could be promising targets for CSC-oriented therapy.

  20. Identification of Lactobacillus plantarum genes modulating the cytokine response of human peripheral blood mononuclear cells

    E-Print Network [OSTI]

    2010-01-01

    effective delivery of probiotics. Ann Rev Food Sci Technol108. 5. Sanders ME: Probiotics: Considerations for humanCL, Gershwin ME: Probiotics and immunity. J Gastroenterol

  1. Adenoviral Mediated Gene Delivery to Human Umbilical Cord Mesenchymal Stromal Cells for Inner Ear Hair Cell Differentiation

    E-Print Network [OSTI]

    Devarajan, Keerthana

    2011-07-28

    . Sensory neural hearing loss (SNHL) is the most common form, which results from degeneration of inner ear sensory hair cells and auditory neurons in the cochlea. In recent years, there has been an increasing interest in gene delivery to mesenchymal stem...

  2. Antiproliferative MCR peptides block physical interaction of insulin with retinoblastoma protein (RB) in human lung cancer cells

    E-Print Network [OSTI]

    Razvan Tudor Radulescu; Kai Kehe

    2007-06-13

    Fifteen years ago, a structural analysis of the hormone insulin and the retinoblastoma tumor suppressor protein (RB) revealed that they may physically interact with one another. Subsequently, an RB peptide corresponding to the proposed RB binding site for insulin was found to recognize full-length insulin in vitro. As part of efforts aimed at developing this RB peptide into an anti-cancer drug, this molecule was chemically coupled to a cellular internalization signal and termed "MCR peptide". Meanwhile, several such MCR peptide variants have been demonstrated to restrain the proliferation of different human cancer cells in vitro and in vivo. Moreover, one of the MCR peptides coined MCR-10 was shown to be capable of interfering with the complex formation between insulin and RB in HepG2 human hepatoma cells, as monitored by immunofluorescence. This latter result indicating an in vivo association between insulin and RB was confirmed by a follow-up study combining the methods of co-immunoprecipitation and immunoblotting. Here, we provide evidence for the existence of the insulin-RB complex in A549 human non-small cell lung cancer cells. Specifically, we demonstrate this heterodimer by means of a magnetic beads-based immunoprecipitation approach and equally show that this dimer can be disrupted by MCR-4 or MCR-10 each of which is known to possess antiproliferative properties, yet to a much lesser extent by a control peptide. Thus, this investigation has yielded another important proof for the occurrence of the insulin-RB dimer and, furthermore, its validity as a target for antineoplastic MCR peptides.

  3. The protein pheromone Er-1 of the ciliate Euplotes raikovi stimulates human T-cell activity: Involvement of interleukin-2 system

    SciTech Connect (OSTI)

    Cervia, Davide; Department of Biomedical and Clinical Sciences, “Luigi Sacco” University Hospital, University of Milan, Milano ; Catalani, Elisabetta; Belardinelli, Maria Cristina; Perrotta, Cristiana; Picchietti, Simona; Alimenti, Claudio; Casini, Giovanni; Fausto, Anna Maria; Vallesi, Adriana

    2013-02-01

    Water-soluble protein signals (pheromones) of the ciliate Euplotes have been supposed to be functional precursors of growth factors and cytokines that regulate cell–cell interaction in multi-cellular eukaryotes. This work provides evidence that native preparations of the Euplotes raikovi pheromone Er-1 (a helical protein of 40 amino acids) specifically increases viability, DNA synthesis, proliferation, and the production of interferon-?, tumor necrosis factor-?, interleukin (IL)-1?, IL-2, and IL-13 in human Jurkat T-cells. Also, Er-1 significantly decreases the mRNA levels of the ? and ? subunits of IL-2 receptor (IL-2R), while the mRNA levels of the ? subunit appeared to be not affected. Jurkat T-cell treatments with Er-1 induced the down-regulation of the IL-2R? subunit by a reversible and time-dependent endocytosis, and increased the levels of phosphorylation of the extracellular signal-regulated kinases (ERK). The cell-type specificity of these effects was supported by the finding that Er-1, although unable to directly influence the growth of human glioma U-373 cells, induced Jurkat cells to synthesize and release factors that, in turn, inhibited the U-373 cell proliferation. Overall, these findings imply that Er-1 coupling to IL-2R and ERK immuno-enhances T-cell activity, and that this effect likely translates to an inhibition of glioma cell growth. -- Highlights: ? Euplotes pheromone Er-1 increases the growth of human Jurkat T-cells. ? Er-1 increases the T-cell production of specific cytokines. ? Er-1 activates interleukin-2 receptor and extracellular signal-regulated kinases. ? The immuno-enhancing effect of Er-1 on Jurkat cells translates to an inhibition of human glioma cell growth.

  4. Potential role of 20S proteasome in maintaining stem cell integrity of human bone marrow stromal cells in prolonged culture expansion

    SciTech Connect (OSTI)

    Lu, Li; Song, Hui-Fang; Zhang, Wei-Guo; Liu, Xue-Qin; Zhu, Qian; Cheng, Xiao-Long; Yang, Gui-Jiao; Li, Ang; Xiao, Zhi-Cheng; Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Melbourne 3800

    2012-05-25

    Highlights: Black-Right-Pointing-Pointer Prolonged culture expansion retards proliferation and induces senescence of hBMSCs. Black-Right-Pointing-Pointer Reduced 20S proteasomal activity and expression potentially contribute to cell aging. Black-Right-Pointing-Pointer MG132-mediated 20S proteasomal inhibition induces senescence-like phenotype. Black-Right-Pointing-Pointer 18{alpha}-GA stimulates proteasomal activity and restores replicative senescence. Black-Right-Pointing-Pointer 18{alpha}-GA retains differentiation without affecting stem cell characterizations. -- Abstract: Human bone marrow stromal cells (hBMSCs) could be used in clinics as precursors of multiple cell lineages following proper induction. Such application is impeded by their characteristically short lifespan, together with the increasing loss of proliferation capability and progressive reduction of differentiation potential after the prolonged culture expansion. In the current study, we addressed the possible role of 20S proteasomes in this process. Consistent with prior reports, long-term in vitro expansion of hBMSCs decreased cell proliferation and increased replicative senescence, accompanied by reduced activity and expression of the catalytic subunits PSMB5 and PSMB1, and the 20S proteasome overall. Application of the proteasome inhibitor MG132 produced a senescence-like phenotype in early passages, whereas treating late-passage cells with 18{alpha}-glycyrrhetinic acid (18{alpha}-GA), an agonist of 20S proteasomes, delayed the senescence progress, enhancing the proliferation and recovering the capability of differentiation. The data demonstrate that activation of 20S proteasomes assists in counteracting replicative senescence of hBMSCs expanded in vitro.

  5. Integrated platform for functional monitoring of biomimetic heart sheets derived from human pluripotent stem cells

    E-Print Network [OSTI]

    Fowlkes, Charless

    of a biomimetic drug screening platform for the early detection of car- diotoxicity is essentialIntegrated platform for functional monitoring of biomimetic heart sheets derived from human present an integrated platform comprised of a biomimetic substrate and physiologically aligned human

  6. Dynamic Skin Triangulation (extended abstract)

    E-Print Network [OSTI]

    Sullivan, John M.

    of physical simulation, where they act as bound- aries of spatial domains that grow and shrink with time stages [1]. Moving bound- aries also arise naturally in mold filling processes, both for metal and other and shrinking. The skin surface is the envelope of this family. Even though the family is infinite, the surface

  7. Differential Glioma-Associated Tumor Antigen Expression Profiles of Human Glioma Cells Grown in Hypoxia

    E-Print Network [OSTI]

    2012-01-01

    were synthesized by Operon Biotech (Huntsville, AL). Themicrobeads (Miltenyi Biotech, Auburn, CA). CD8+ T cells (rabbit HIF-1a (Santa Crux Biotech, Santa Cruz, CA), or the

  8. Soft inertial microfluidics for high throughput separation of bacteria from human blood cells

    E-Print Network [OSTI]

    Wu, Zhigang

    2009-01-01

    Soft inertial microfluidics for high throughput separation1 Introduction Microfluidics has gained significant advancesof mammalian cells using microfluidics 3,4 , there have been

  9. Nanofibrous Electrospun Polymers for Reprogramming Human Cells TRAVIS CORDIE,1,2

    E-Print Network [OSTI]

    Saha, Krishanu

    oriented and aligned structures was prepared by electrospinning four polymers [polylactic acid (PLA), polyc cells, Reprogramming, Bioma- terials, Electrospinning, Nuclear shape. INTRODUCTION Dramatic changes

  10. Low-dose responses to 2,3,7,8-tetrachlorodibenzo-p-dioxin in single living human cells measured by synchrotron infrared spectromicroscopy

    E-Print Network [OSTI]

    Low-dose responses to 2,3,7,8-tetrachlorodibenzo-p-dioxin in single living human cells measured, and reproductive defects in animals1-6 and humans.7-14 Among this family of pollutants, 2,3,7,8-tetrachlorodibenzo-p-dioxin

  11. Targeting miR-21 enhances the sensitivity of human colon cancer HT-29 cells to chemoradiotherapy in vitro

    SciTech Connect (OSTI)

    Deng, Jun; Lei, Wan; Fu, Jian-Chun; Zhang, Ling; Li, Jun-He; Xiong, Jian-Ping

    2014-01-17

    Highlight: •MiR-21 plays a significant role in 5-FU resistance. •This role might be attributed to targeting of hMSH2 as well as TP and DPD via miR-21 targeted hMSH2. •Indirectly targeted TP and DPD to influence 5-FU chemotherapy sensitivity. -- Abstract: 5-Fluorouracil (5-FU) is a classic chemotherapeutic drug that has been widely used for colorectal cancer treatment, but colorectal cancer cells are often resistant to primary or acquired 5-FU therapy. Several studies have shown that miR-21 is significantly elevated in colorectal cancer. This suggests that this miRNA might play a role in this resistance. In this study, we investigated this possibility and the possible mechanism underlying this role. We showed that forced expression of miR-21 significantly inhibited apoptosis, enhanced cell proliferation, invasion, and colony formation ability, promoted G1/S cell cycle transition and increased the resistance of tumor cells to 5-FU and X radiation in HT-29 colon cancer cells. Furthermore, knockdown of miR-21 reversed these effects on HT-29 cells and increased the sensitivity of HT-29/5-FU to 5-FU chemotherapy. Finally, we showed that miR-21 targeted the human mutS homolog2 (hMSH2), and indirectly regulated the expression of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD). These results demonstrate that miR-21 may play an important role in the 5-FU resistance of colon cancer cells.

  12. Turbine vane with high temperature capable skins

    DOE Patents [OSTI]

    Morrison, Jay A. (Oviedo, FL)

    2012-07-10

    A turbine vane assembly includes an airfoil extending between an inner shroud and an outer shroud. The airfoil can include a substructure having an outer peripheral surface. At least a portion of the outer peripheral surface is covered by an external skin. The external skin can be made of a high temperature capable material, such as oxide dispersion strengthened alloys, intermetallic alloys, ceramic matrix composites or refractory alloys. The external skin can be formed, and the airfoil can be subsequently bi-cast around or onto the skin. The skin and the substructure can be attached by a plurality of attachment members extending between the skin and the substructure. The skin can be spaced from the outer peripheral surface of the substructure such that a cavity is formed therebetween. Coolant can be supplied to the cavity. Skins can also be applied to the gas path faces of the inner and outer shrouds.

  13. Active skin for turbulent drag reduction 

    E-Print Network [OSTI]

    Mani, Raghavendran

    2002-01-01

    capitalizes on recent advances in active turbulent drag reduction and active material based actuation to develop an active or "smart" skin for turbulent drag reduction in realistic flight conditions. The skin operation principle is based on computational...

  14. Tumor exosomes induce tunneling nanotubes in lipid raft-enriched regions of human mesothelioma cells

    SciTech Connect (OSTI)

    Thayanithy, Venugopal; Babatunde, Victor; Dickson, Elizabeth L.; Wong, Phillip; Oh, Sanghoon; Ke, Xu; Barlas, Afsar; Fujisawa, Sho; Romin, Yevgeniy; Moreira, André L.; Downey, Robert J.; Steer, Clifford J.; Subramanian, Subbaya; Manova-Todorova, Katia; Moore, Malcolm A.S.; Lou, Emil

    2014-04-15

    Tunneling nanotubes (TnTs) are long, non-adherent, actin-based cellular extensions that act as conduits for transport of cellular cargo between connected cells. The mechanisms of nanotube formation and the effects of the tumor microenvironment and cellular signals on TnT formation are unknown. In the present study, we explored exosomes as potential mediators of TnT formation in mesothelioma and the potential relationship of lipid rafts to TnT formation. Mesothelioma cells co-cultured with exogenous mesothelioma-derived exosomes formed more TnTs than cells cultured without exosomes within 24–48 h; and this effect was most prominent in media conditions (low-serum, hyperglycemic medium) that support TnT formation (1.3–1.9-fold difference). Fluorescence and electron microscopy confirmed the purity of isolated exosomes and revealed that they localized predominantly at the base of and within TnTs, in addition to the extracellular environment. Time-lapse microscopic imaging demonstrated uptake of tumor exosomes by TnTs, which facilitated intercellular transfer of these exosomes between connected cells. Mesothelioma cells connected via TnTs were also significantly enriched for lipid rafts at nearly a 2-fold higher number compared with cells not connected by TnTs. Our findings provide supportive evidence of exosomes as potential chemotactic stimuli for TnT formation, and also lipid raft formation as a potential biomarker for TnT-forming cells. - Highlights: • Exosomes derived from malignant cells can stimulate an increased rate in the formation of tunneling nanotubes. • Tunneling nanotubes can serve as conduits for intercellular transfer of these exosomes. • Most notably, exosomes derived from benign mesothelial cells had no effect on nanotube formation. • Cells forming nanotubes were enriched in lipid rafts at a greater number compared with cells not forming nanotubes. • Our findings suggest causal and potentially synergistic association of exosomes and tunneling nanotubes in cancer.

  15. 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.

  16. Latent infection of myeloid progenitors by human cytomegalovirus protects cells from FAS-mediated apoptosis through the cellular IL-10/PEA15 pathway

    E-Print Network [OSTI]

    Poole, Emma; Lau, Jonathan; Sinclair, John

    2015-05-08

    ., Breidenstein, A. & Compton, T. (2012). Human cytomegalovirus activation of ERK 214 and myeloid cell leukemia-1 protein correlates with survival of latently infected cells. Proc 215 Natl Acad Sci U S A 109, 588-593. 216 Reeves, M. B., MacAry, P. A., Lehner, P...

  17. Discovery of Human sORF-Encoded Polypeptides (SEPs) in Cell Lines and Tissue

    E-Print Network [OSTI]

    Ma, Jiao

    The existence of nonannotated protein-coding human short open reading frames (sORFs) has been revealed through the direct detection of their sORF-encoded polypeptide (SEP) products. The discovery of novel SEPs increases ...

  18. Quantitative analysis of cell composition and purity of human pancreatic islet preparations

    E-Print Network [OSTI]

    Pisania, Anna

    Despite improvements in outcomes for human islet transplantation, characterization of islet preparations remains poorly defined. This study used both light microscopy (LM) and electron microscopy (EM) to characterize 33 ...

  19. Common Genetic Variants Modulate Pathogen-Sensing Responses in Human Dendritic Cells

    E-Print Network [OSTI]

    Lee, M. N.

    Little is known about how human genetic variation affects the responses to environmental stimuli in the context of complex diseases. Experimental and computational approaches were applied to determine the effects of genetic ...

  20. Leptin upregulates telomerase activity and transcription of human telomerase reverse transcriptase in MCF-7 breast cancer cells

    SciTech Connect (OSTI)

    Ren, He; Zhao, Tiansuo; Wang, Xiuchao; Gao, Chuntao; Wang, Jian; Yu, Ming; Hao, Jihui

    2010-03-26

    The aim was to analyze the mechanism of leptin-induced activity of telomerase in MCF-7 breast cancer cells. We found that leptin activated telomerase in a dose-dependent manner; leptin upregulated the expression of Human Telomerase Reverse Transcriptase (hTERT) at mRNA and protein levels; blockade of signal transducer and activator of transcription 3 (STAT3) phosphorylation significantly counteracted leptin-induced hTERT transcription and protein expression; chromatin immunoprecipitation analysis showed that leptin enhanced the binding of STAT3 to the hTERT promoter. This study uncovers a new mechanism of the proliferative effect of leptin on breast cancer cells and provides a new explanation of obesity-related breast cancer.

  1. Oncogenic Radiation Abscopal Effects In Vivo: Interrogating Mouse Skin

    SciTech Connect (OSTI)

    Mancuso, Mariateresa, E-mail: mariateresa.mancuso@enea.it [Laboratory of Radiation Biology and Biomedicine, Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile (ENEA), Casaccia Research Centre, Rome (Italy); Leonardi, Simona [Laboratory of Radiation Biology and Biomedicine, Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile (ENEA), Casaccia Research Centre, Rome (Italy); Giardullo, Paola; Pasquali, Emanuela [Department of Radiation Physics, Guglielmo Marconi University, Rome (Italy); Tanori, Mirella [Laboratory of Radiation Biology and Biomedicine, Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile (ENEA), Casaccia Research Centre, Rome (Italy); De Stefano, Ilaria [Department of Radiation Physics, Guglielmo Marconi University, Rome (Italy); Casciati, Arianna [Laboratory of Radiation Biology and Biomedicine, Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile (ENEA), Casaccia Research Centre, Rome (Italy); Naus, Christian C. [Department of Cellular and Physiological Sciences, The Life Sciences Institute, University of British Columbia, Vancouver, British Columbia (Canada); Pazzaglia, Simonetta; Saran, Anna [Laboratory of Radiation Biology and Biomedicine, Agenzia Nazionale per le Nuove Tecnologie, l'Energia e lo Sviluppo Economico Sostenibile (ENEA), Casaccia Research Centre, Rome (Italy)

    2013-08-01

    Purpose: To investigate the tissue dependence in transmission of abscopal radiation signals and their oncogenic consequences in a radiosensitive mouse model and to explore the involvement of gap junction intercellular communication (GJIC) in mediating radiation tumorigenesis in off-target mouse skin. Methods and Materials: Patched1 heterozygous (Ptch1{sup +/?}) mice were irradiated at postnatal day 2 (P2) with 10 Gy of x-rays. Individual lead cylinders were used to protect the anterior two-thirds of the body, whereas the hindmost part was directly exposed to radiation. To test the role of GJICs and their major constituent connexin43 (Cx43), crosses between Ptch1{sup +/?} and Cx43{sup +/?} mice were similarly irradiated. These mouse groups were monitored for their lifetime, and skin basal cell carcinomas (BCCs) were counted and recorded. Early responses to DNA damage - Double Strand Breaks (DSBs) and apoptosis - were also evaluated in shielded and directly irradiated skin areas. Results: We report abscopal tumor induction in the shielded skin of Ptch1{sup +/?} mice after partial-body irradiation. Endpoints were induction of early nodular BCC-like tumors and macroscopic infiltrative BCCs. Abscopal tumorigenesis was significantly modulated by Cx43 status, namely, Cx43 reduction was associated with decreased levels of DNA damage and oncogenesis in out-of-field skin, suggesting a key role of GJIC in transmission of oncogenic radiation signals to unhit skin. Conclusions: Our results further characterize the nature of abscopal responses and the implications they have on pathologic processes in different tissues, including their possible underlying mechanistic bases.

  2. Effects of FGF-2 on human adipose tissue derived adult stem cells morphology and chondrogenesis enhancement in Transwell culture

    SciTech Connect (OSTI)

    Kabiri, Azadeh; Esfandiari, Ebrahim; Hashemibeni, Batool; Kazemi, Mohammad; Mardani, Mohammad; Esmaeili, Abolghasem

    2012-07-27

    Highlights: Black-Right-Pointing-Pointer We investigated effects of FGF-2 on hADSCs. Black-Right-Pointing-Pointer We examine changes in the level of gene expressions of SOX-9, aggrecan and collagen type II and type X. Black-Right-Pointing-Pointer FGF-2 induces chondrogenesis in hADSCs, which Bullet Increasing information will decrease quality if hospital costs are very different. Black-Right-Pointing-Pointer The result of this study may be beneficial in cartilage tissue engineering. -- Abstract: Injured cartilage is difficult to repair due to its poor vascularisation. Cell based therapies may serve as tools to more effectively regenerate defective cartilage. Both adult mesenchymal stem cells (MSCs) and human adipose derived stem cells (hADSCs) are regarded as potential stem cell sources able to generate functional cartilage for cell transplantation. Growth factors, in particular the TGF-b superfamily, influence many processes during cartilage formation, including cell proliferation, extracellular matrix synthesis, maintenance of the differentiated phenotype, and induction of MSCs towards chondrogenesis. In the current study, we investigated the effects of FGF-2 on hADSC morphology and chondrogenesis in Transwell culture. hADSCs were obtained from patients undergoing elective surgery, and then cultured in expansion medium alone or in the presence of FGF-2 (10 ng/ml). mRNA expression levels of SOX-9, aggrecan and collagen type II and type X were quantified by real-time polymerase chain reaction. The morphology, doubling time, trypsinization time and chondrogenesis of hADSCs were also studied. Expression levels of SOX-9, collagen type II, and aggrecan were all significantly increased in hADSCs expanded in presence of FGF-2. Furthermore FGF-2 induced a slender morphology, whereas doubling time and trypsinization time decreased. Our results suggest that FGF-2 induces hADSCs chondrogenesis in Transwell culture, which may be beneficial in cartilage tissue engineering.

  3. 5-Methoxyflavanone induces cell cycle arrest at the G2/M phase, apoptosis and autophagy in HCT116 human colon cancer cells

    SciTech Connect (OSTI)

    Shin, Soon Young [SMART Institute of Advanced Biomedical Science, Konkuk University Medical Center, Seoul 143-701 (Korea, Republic of); Department of Biomedical Science and Technology, Research Center for Transcription Control, Konkuk University, Seoul 143-701 (Korea, Republic of); Hyun, Jiye [Division of Bioscience and Biotechnology, BMIC, Konkuk University, Seoul 143-701 (Korea, Republic of); Yu, Jae-Ran [Department of Environmental and Tropical Medicine, Konkuk University School of Medicine, Seoul 143-701 (Korea, Republic of); Lim, Yoongho, E-mail: yoongho@konkuk.ac.kr [Division of Bioscience and Biotechnology, BMIC, Konkuk University, Seoul 143-701 (Korea, Republic of); Lee, Young Han, E-mail: yhlee58@konkuk.ac.kr [SMART Institute of Advanced Biomedical Science, Konkuk University Medical Center, Seoul 143-701 (Korea, Republic of); Department of Biomedical Science and Technology, Research Center for Transcription Control, Konkuk University, Seoul 143-701 (Korea, Republic of)

    2011-08-01

    Natural flavonoids have diverse pharmacological activities, including anti-oxidative, anti-inflammatory, and anti-cancer activities. In this study, we investigated the molecular mechanism underlying the action of 5-methoxyflavanone (5-MF) which has a strong bioavailability and metabolic stability. Our results show that 5-MF inhibited the growth and clonogenicity of HCT116 human colon cancer cells, and that it activated DNA damage responses, as revealed by the accumulation of p53 and the phosphorylation of DNA damage-sensitive proteins, including ataxia-telangiectasia mutated (ATM) at Ser1981, checkpoint kinase 2 (Chk2) at Thr68, and histone H2AX at Ser139. 5-MF-induced DNA damage was confirmed in a comet tail assay. We also found that 5-MF increased the cleavage of caspase-2 and -7, leading to the induction of apoptosis. Pretreatment with the ATM inhibitor KU55933 enhanced 5-MF-induced {gamma}-H2AX formation and caspase-7 cleavage. HCT116 cells lacking p53 (p53{sup -/-}) or p21 (p21{sup -/-}) exhibited increased sensitivity to 5-MF compared to wild-type cells. 5-MF further induced autophagy via an ERK signaling pathway. Blockage of autophagy with the MEK inhibitor U0126 potentiated 5-MF-induced {gamma}-H2AX formation and caspase-2 activation. These results suggest that a caspase-2 cascade mediates 5-MF-induced anti-tumor activity, while an ATM/Chk2/p53/p21 checkpoint pathway and ERK-mediated autophagy act as a survival program to block caspase-2-mediated apoptosis induced by 5-MF. - Graphical abstract: Display Omitted Highlights: > 5-MF inhibits the proliferation of HCT116 colon cancer cells. > 5-MF inhibits cell cycle progression and induces apoptosis. > Inhibition of autophagy triggers 5-MF-induced apoptosis. > Inhibition of ERK signaling blocks 5-MF-induced autophagy but activates apoptosis. > Treatment with 5-MF in combination with an ERK inhibitor may be a potential therapeutic strategy in human colon cancer.

  4. Relationship between beta-cell mass and fasting blood glucose concentration in humans

    E-Print Network [OSTI]

    Ritzel, Robert A; Butler, Alexandra E; Rizza, Robert A; Veldhuis, Johannes D; Butler, Peter C

    2006-01-01

    Between ?-Cell Mass and Fasting Blood Glucose Concentrationnondiabetic, had impaired fasting glu- cose, or had type 26.9 kg/m 2 ), had impaired fasting glucose (?110 mg/dl [6.1

  5. High-throughput gene expression profiling of memory differentiation in primary human T cells

    E-Print Network [OSTI]

    Angelosanto, Jill

    Background: The differentiation of naive T and B cells into memory lymphocytes is essential for immunity to pathogens. Therapeutic manipulation of this cellular differentiation program could improve vaccine efficacy and ...

  6. In Vitro Properties of Human Mesenchymal Stem Cells and Their Ability to Modulate Inflammation in Vivo 

    E-Print Network [OSTI]

    Bazhanov, Nikolay

    2015-05-01

    microdissection was used to isolate distinct inside and outside regions of the colony for RNA isolation. When assayed by microarray the cells from inside and outside regions of the colony showed distinct patterns of gene expression profile. In functional studies...

  7. Investigation of in-vivo skin autofluorescence lifetimes under long-term cw optical excitation

    SciTech Connect (OSTI)

    Lihachev, A; Ferulova, I; Vasiljeva, K; Spigulis, J

    2014-08-31

    The main results obtained during the last five years in the field of laser-excited in-vivo human skin photobleaching effects are presented. The main achievements and results obtained, as well as methods and experimental devices are briefly described. In addition, the impact of long-term 405-nm cw low-power laser excitation on the skin autofluorescence lifetime is experimentally investigated. (laser biophotonics)

  8. Nickel compounds induce apoptosis in human bronchial epithelial Beas-2B cells by activation of c-Myc through ERK pathway

    SciTech Connect (OSTI)

    Li Qin; Suen, T.-C.; Sun Hong; Arita, Adriana [New York University School of Medicine, Nelson Institute of Environmental Medicine, 57 Old Forge Road, NY 10987 (United States); Costa, Max [New York University School of Medicine, Nelson Institute of Environmental Medicine, 57 Old Forge Road, NY 10987 (United States)], E-mail: max.costa@nyumc.org

    2009-03-01

    Nickel compounds are carcinogenic to humans and have been shown to alter epigenetic homeostasis. The c-Myc protein controls 15% of human genes and it has been shown that fluctuations of c-Myc protein alter global epigenetic marks. Therefore, the regulation of c-Myc by nickel ions in immortalized but not tumorigenic human bronchial epithelial Beas-2B cells was examined in this study. It was found that c-Myc protein expression was increased by nickel ions in non-tumorigenic Beas-2B and human keratinocyte HaCaT cells. The results also indicated that nickel ions induced apoptosis in Beas-2B cells. Knockout of c-Myc and its restoration in a rat cell system confirmed the essential role of c-Myc in nickel ion-induced apoptosis. Further studies in Beas-2B cells showed that nickel ion increased the c-Myc mRNA level and c-Myc promoter activity, but did not increase c-Myc mRNA and protein stability. Moreover, nickel ion upregulated c-Myc in Beas-2B cells through the MEK/ERK pathway. Collectively, the results demonstrate that c-Myc induction by nickel ions occurs via an ERK-dependent pathway and plays a crucial role in nickel-induced apoptosis in Beas-2B cells.

  9. Specific expression of the human voltage-gated proton channel Hv1 in highly metastatic breast cancer cells, promotes tumor progression and metastasis

    SciTech Connect (OSTI)

    Wang, Yifan [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China)] [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China); Li, Shu Jie, E-mail: shujieli@nankai.edu.cn [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China); Pan, Juncheng [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China)] [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China); Che, Yongzhe, E-mail: cheli@nankai.edu.cn [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Medicine, Nankai University, Tianjin 300071 (China)] [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Medicine, Nankai University, Tianjin 300071 (China); Yin, Jian [Cancer Institute and Hospital, Tianjin Medical University, Tianjin 300060 (China)] [Cancer Institute and Hospital, Tianjin Medical University, Tianjin 300060 (China); Zhao, Qing [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China)] [The Key Laboratory of Bioactive Materials, Ministry of Education, School of Physics Science, Nankai University, Tianjin 300071 (China)

    2011-08-26

    Highlights: {yields} Hv1 is specifically expressed in highly metastatic human breast tumor tissues. {yields} Hv1 regulates breast cancer cytosolic pH. {yields} Hv1 acidifies extracellular milieu. {yields} Hv1 exacerbates the migratory ability of metastatic cells. -- Abstract: The newly discovered human voltage-gated proton channel Hv1 is essential for proton transfer, which contains a voltage sensor domain (VSD) without a pore domain. We report here for the first time that Hv1 is specifically expressed in the highly metastatic human breast tumor tissues, but not in poorly metastatic breast cancer tissues, detected by immunohistochemistry. Meanwhile, real-time RT-PCR and immunocytochemistry showed that the expression levels of Hv1 have significant differences among breast cancer cell lines, MCF-7, MDA-MB-231, MDA-MB-468, MDA-MB-453, T-47D and SK-BR-3, in which Hv1 is expressed at a high level in highly metastatic human breast cancer cell line MDA-MB-231, but at a very low level in poorly metastatic human breast cancer cell line MCF-7. Inhibition of Hv1 expression in the highly metastatic MDA-MB-231 cells by small interfering RNA (siRNA) significantly decreases the invasion and migration of the cells. The intracellular pH of MDA-MB-231 cells down-regulated Hv1 expression by siRNA is obviously decreased compared with MDA-MB-231 with the scrambled siRNA. The expression of matrix metalloproteinase-2 and gelatinase activity in MDA-MB-231 cells suppressed Hv1 by siRNA were reduced. Our results strongly suggest that Hv1 regulates breast cancer intracellular pH and exacerbates the migratory ability of metastatic cells.

  10. Encapsulated human hepatocellular carcinoma cells by alginate gel beads as an in vitro metastasis model

    SciTech Connect (OSTI)

    Xu, Xiao-xi; Liu, Chang [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); University of Chinese Academy of Sciences, 19A Yuquanlu, Beijing 100049 (China); Liu, Yang [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Li, Nan [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); University of Chinese Academy of Sciences, 19A Yuquanlu, Beijing 100049 (China); Guo, Xin [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Wang, Shu-jun [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); School of Life Science and Biotechnology, Dalian University of Technology, 2 Linggong Road, Dalian 116024 (China); Sun, Guang-wei, E-mail: sungw@dicp.ac.cn [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Wang, Wei [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Ma, Xiao-jun, E-mail: maxj@dicp.ac.cn [Laboratory of Biomedical Material Engineering, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China)

    2013-08-15

    Hepatocellular carcinoma (HCC) is the most common primary liver cancer and often forms metastases, which are the most important prognostic factors. For further elucidation of the mechanism underlying the progression and metastasis of HCC, a culture system mimicking the in vivo tumor microenvironment is needed. In this study, we investigated the metastatic ability of HCC cells cultured within alginate gel (ALG) beads. In the culture system, HCC cells formed spheroids by proliferation and maintained in nuclear abnormalities. The gene and protein expression of metastasis-related molecules was increased in ALG beads, compared with the traditional adhesion culture. Furthermore, several gene expression levels in ALG bead culture system were even closer to liver cancer tissues. More importantly, in vitro invasion assay showed that the invasion cells derived from ALG beads was 7.8-fold higher than adhesion cells. Our results indicated that the in vitro three-dimensional (3D) model based on ALG beads increased metastatic ability compared with adhesion culture, even partly mimicked the in vivo tumor tissues. Moreover, due to the controllable preparation conditions, steady characteristics and production at large-scale, the 3D ALG bead model would become an important tool used in the high-throughput screening of anti-metastasis drugs and the metastatic mechanism research. -- Highlights: •We established a 3D metastasis model mimicking the metastatic ability in vivo. •The invasion ability of cells derived from our model was increased significantly. •The model is easy to reproduce, convenient to handle, and amenable for large-scale.

  11. 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.

  12. JAB1 regulates unphosphorylated STAT3 DNA-binding activity through protein–protein interaction in human colon cancer cells

    SciTech Connect (OSTI)

    Nishimoto, Arata, E-mail: anishimo@yamaguchi-u.ac.jp [Department of Surgery and Clinical Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505 (Japan)] [Department of Surgery and Clinical Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505 (Japan); Kugimiya, Naruji; Hosoyama, Toru; Enoki, Tadahiko [Department of Surgery and Clinical Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505 (Japan)] [Department of Surgery and Clinical Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505 (Japan); Li, Tao-Sheng [Department of Stem Cell Biology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523 (Japan)] [Department of Stem Cell Biology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523 (Japan); Hamano, Kimikazu [Department of Surgery and Clinical Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505 (Japan)] [Department of Surgery and Clinical Science, Yamaguchi University Graduate School of Medicine, 1-1-1 Minami-Kogushi, Ube, Yamaguchi 755-8505 (Japan)

    2013-08-30

    Highlights: •JAB1 interacted with unphosphorylated STAT3 in the nucleus. •JAB1 knockdown tended to increase nuclear STAT3 expression. •JAB1 knockdown significantly decreased unphosphorylated STAT3 DNA-binding activity. •JAB1 knockdown significantly decreased MDR1, NANOG, and VEGF expressions. •Nuclear JAB1, but not nuclear STAT3, correlated with STAT3 DNA-binding activity. -- Abstract: Recent studies have revealed that unphosphorylated STAT3 forms a dimer, translocates to the nucleus, binds to the STAT3 binding site, and activates the transcription of STAT3 target genes, thereby playing an important role in oncogenesis in addition to phosphorylated STAT3. Among signaling steps of unphosphorylated STAT3, nuclear translocation and target DNA-binding are the critical steps for its activation. Therefore, elucidating the regulatory mechanism of these signaling steps of unphosphorylated STAT3 is a potential step in the discovery of a novel cancer drug. However, the mechanism of unphosphorylated STAT3 binding to the promoter of target genes remains unclear. In this study, we focused on Jun activation domain-binding protein 1 (JAB1) as a candidate protein that regulates unphosphorylated STAT3 DNA-binding activity. Initially, we observed that both unphosphorylated STAT3 and JAB1 existed in the nucleus of human colon cancer cell line COLO205 at the basal state (no cytokine stimulation). On the other hand, phosphorylated STAT3 did not exist in the nucleus of COLO205 cells at the basal state. Immunoprecipitation using nuclear extract of COLO205 cells revealed that JAB1 interacted with unphosphorylated STAT3. To investigate the effect of JAB1 on unphosphorylated STAT3 activity, RNAi studies were performed. Although JAB1 knockdown tended to increase nuclear STAT3 expression, it significantly decreased unphosphorylated STAT3 DNA-binding activity. Subsequently, JAB1 knockdown significantly decreased the expression levels of MDR1, NANOG, and VEGF, which are STAT3 target genes. Furthermore, the expression level of nuclear JAB1, but not nuclear STAT3, correlated with unphosphorylated STAT3 DNA-binding activity between COLO205 and LoVo cells. Taken together, these results suggest that nuclear JAB1 positively regulates unphosphorylated STAT3 DNA-binding activity through protein–protein interaction in human colon cancer cell line COLO205.

  13. Assessment of beating parameters in human induced pluripotent stem cells enables quantitative in vitro screening for cardiotoxicity

    SciTech Connect (OSTI)

    Sirenko, Oksana; Cromwell, Evan F.; Crittenden, Carole; Wignall, Jessica A.; Wright, Fred A.; Rusyn, Ivan

    2013-12-15

    Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes show promise for screening during early drug development. Here, we tested a hypothesis that in vitro assessment of multiple cardiomyocyte physiological parameters enables predictive and mechanistically-interpretable evaluation of cardiotoxicity in a high-throughput format. Human iPSC-derived cardiomyocytes were exposed for 30 min or 24 h to 131 drugs, positive (107) and negative (24) for in vivo cardiotoxicity, in up to 6 concentrations (3 nM to 30 uM) in 384-well plates. Fast kinetic imaging was used to monitor changes in cardiomyocyte function using intracellular Ca{sup 2+} flux readouts synchronous with beating, and cell viability. A number of physiological parameters of cardiomyocyte beating, such as beat rate, peak shape (amplitude, width, raise, decay, etc.) and regularity were collected using automated data analysis. Concentration–response profiles were evaluated using logistic modeling to derive a benchmark concentration (BMC) point-of-departure value, based on one standard deviation departure from the estimated baseline in vehicle (0.3% dimethyl sulfoxide)-treated cells. BMC values were used for cardiotoxicity classification and ranking of compounds. Beat rate and several peak shape parameters were found to be good predictors, while cell viability had poor classification accuracy. In addition, we applied the Toxicological Prioritization Index (ToxPi) approach to integrate and display data across many collected parameters, to derive “cardiosafety” ranking of tested compounds. Multi-parameter screening of beating profiles allows for cardiotoxicity risk assessment and identification of specific patterns defining mechanism-specific effects. These data and analysis methods may be used widely for compound screening and early safety evaluation in drug development. - Highlights: • Induced pluripotent stem cell-derived cardiomyocytes are promising in vitro models. • We tested if evaluation of cardiotoxicity is possible in a high-throughput format. • The assay shows benefits of automated data integration across multiple parameters. • Quantitative assessment of concentration–response is possible using iPSCs. • Multi-parametric screening allows for cardiotoxicity risk assessment.

  14. Effects of drugs in subtoxic concentrations on the metabolic fluxes in human hepatoma cell line Hep G2

    SciTech Connect (OSTI)

    Niklas, Jens [Biochemical Engineering Institute, Saarland University, Campus A1.5, D-66123 Saarbruecken (Germany); Noor, Fozia, E-mail: fozia.noor@mx.uni-saarland.d [Biochemical Engineering Institute, Saarland University, Campus A1.5, D-66123 Saarbruecken (Germany); Heinzle, Elmar [Biochemical Engineering Institute, Saarland University, Campus A1.5, D-66123 Saarbruecken (Germany)

    2009-11-01

    Commonly used cytotoxicity assays assess the toxicity of a compound by measuring certain parameters which directly or indirectly correlate to the viability of the cells. However, the effects of a given compound at concentrations considerably below EC{sub 50} values are usually not evaluated. These subtoxic effects are difficult to identify but may eventually cause severe and costly long term problems such as idiosyncratic hepatotoxicity. We determined the toxicity of three hepatotoxic compounds, namely amiodarone, diclofenac and tacrine on the human hepatoma cell line Hep G2 using an online kinetic respiration assay and analysed the effects of subtoxic concentrations of these drugs on the cellular metabolism by using metabolic flux analysis. Several changes in the metabolism could be detected upon exposure to subtoxic concentrations of the test compounds. Upon exposure to diclofenac and tacrine an increase in the TCA-cycle activity was observed which could be a signature of an uncoupling of the oxidative phosphorylation. The results indicate that metabolic flux analysis could serve as an invaluable novel tool for the investigation of the effects of drugs. The described methodology enables tracking the toxicity of compounds dynamically using the respiration assay in a range of concentrations and the metabolic flux analysis permits interesting insights into the changes in the central metabolism of the cell upon exposure to drugs.

  15. SOX17 is a Critical Specifier of Human Primordial Germ Cell Fate

    E-Print Network [OSTI]

    Irie, Naoko; Weinberger, Leehee; Tang, Walfred W. C.; Kobayashi, Toshihiro; Viukov, Sergey; Manor, Yair S.; Dietmann, Sabine; Hanna, Jacob H.; Surani, M. Azim

    2014-12-24

    cells/ 12-well were plated on vitro- nectin/gelatin-coated plates (Gafni et al., 2013) in N2B27 medium (Ying et al., 2008) with 1% KSR, 10 ng/ml bFGF (SCI), 1 ng/ml TGF-b1 (Peprotech), or 20 ng/ml Activin A (SCI) and 10 mM ROCK inhibitor. Medium...

  16. Regulation of human hepatocellular carcinoma cells by Spred2 and correlative studies on its mechanism

    SciTech Connect (OSTI)

    Ma, Xiao-Ni [Lanzhou University of Technology, Lanzhou 730050 (China)] [Lanzhou University of Technology, Lanzhou 730050 (China); Liu, Xiao-Yun [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China) [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China); Center for Disease Control and Prevention, Lanzhou Military Command, Lanzhou 730020 (China); Yang, Yue-Feng [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China)] [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China); Xiao, Feng-Jun [Lanzhou University of Technology, Lanzhou 730050 (China)] [Lanzhou University of Technology, Lanzhou 730050 (China); Li, Qing-Fang [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China)] [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China); Yan, Jun [Lanzhou University of Technology, Lanzhou 730050 (China)] [Lanzhou University of Technology, Lanzhou 730050 (China); Zhang, Qun-Wei; Wang, Li-Sheng [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China)] [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China); Li, Xue-Yan, E-mail: llglixueyan@163.com [Lanzhou University of Technology, Lanzhou 730050 (China)] [Lanzhou University of Technology, Lanzhou 730050 (China); Wang, Hua, E-mail: wanghua@bmi.ac.cn [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China)] [Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing 100850 (China)

    2011-07-15

    Highlights: {yields} Hepatocellular carcinoma is inhibited by Spred2 through as yet unclear mechanisms. {yields} We studied the overexpression of Spred2 in cell line and murine tumor models of HCC. {yields} Spred2 inhibited cell proliferation and migration via attenuating ERK signaling. {yields} Spred2 overexpression induced apoptosis via caspase-3 and downregulated Mcl-1. {yields} A Spred2 knockdown markedly induced tumor growth in vivo. -- Abstract: Members of the Spred gene family are negative regulators of the Ras/Raf-1/ERK pathway, which has been associated with several features of the tumor malignancy. However, the effect of Spred genes on hepatocellular carcinoma (HCC) remains uninvestigated. In the present work, we analyzed the in vitro and in vivo effects of Spred2 expression on the hepatic carcinoma cell line, SMMC-7721. In addition to attenuated ERK activation, which inhibited the proliferation and migration of unstimulated and HGF-stimulated SMMC-7721 cells. Adenovirus-mediated Spred2 overexpression induced the activation of caspase-3 and apoptosis, as well as reduced the expression level of Mcl-1. Most importantly, the knockdown of Spred2 markedly enhanced tumor growth in vivo. In conclusion, these results suggest that Spred2 could qualify as a potential therapeutic target in HCC.

  17. Multi-kilobase homozygous targeted gene replacement in human induced pluripotent stem cells

    E-Print Network [OSTI]

    Byrne, SM; Ortiz, L; Mali, P; Aach, J; Church, GM

    2015-01-01

    110, 5564–5569. 27. Aach,J. , Mali,P. and Church,G.M. (2014)J.L. , De Los Angeles,A. , Mali,P. , Aach,J. , Kim-Kiselak,Mol. Cell Biol. , 14, 19. Mali,P. , Yang,L. , Esvelt,K.M. ,

  18. Stationary turbine component with laminated skin

    DOE Patents [OSTI]

    James, Allister W. (Orlando, FL)

    2012-08-14

    A stationary turbine engine component, such as a turbine vane, includes a internal spar and an external skin. The internal spar is made of a plurality of spar laminates, and the external skin is made of a plurality of skin laminates. The plurality of skin laminates interlockingly engage the plurality of spar laminates such that the external skin is located and held in place. This arrangement allows alternative high temperature materials to be used on turbine engine components in areas where their properties are needed without having to make the entire component out of such material. Thus, the manufacturing difficulties associated with making an entire component of such a material and the attendant high costs are avoided. The skin laminates can be made of advanced generation single crystal superalloys, intermetallics and refractory alloys.

  19. Chondrogenic differentiation of stem cells in human umbilical cord stroma with PGA and PLLA scaffolds

    E-Print Network [OSTI]

    Zhao, Liang; Detamore, Michael S.

    2015-01-01

    , and en- dothelial cells [8-15]. Recently, our laboratory has suc- cessfully induced hUCMSCs into osteogenic and chon- drogenic lineages [14-16]. For engineering articular car- tilage implants, a crucial consideration is the scaffolding biomaterial.... These biomaterials have included a variety of natural gels and hydrogels based on collagen, glyco- saminoglycans, hyaluronic acid, agarose, alginate and gelatin [17-22], as well as a number of synthetic materi- als used as scaffolds for chondrogenic differentiation...

  20. Melatonin Protects Human Cells from Clustered DNA Damages, Killing and Acquisition of Soft Agar Growth Induced by X-rays or 970 MeV/n Fe ions

    SciTech Connect (OSTI)

    Das, B.; Sutherland, B.; Bennett, P. V.; Cutter, N. C.; Sutherland, J. C.

    2011-06-01

    We tested the ability of melatonin (N-acetyl-5 methoxytryptamine), a highly effective radical scavenger and human hormone, to protect DNA in solution and in human cells against induction of complex DNA clusters and biological damage induced by low or high linear energy transfer radiation (100 kVp X-rays, 970 MeV/nucleon Fe ions). Plasmid DNA in solution was treated with increasing concentrations of melatonin (0.0-3.5 mM) and were irradiated with X-rays. Human cells (28SC monocytes) were also irradiated with X-rays and Fe ions with and without 2 mM melatonin. Agarose plugs containing genomic DNA were subjected to Contour Clamped Homogeneous Electrophoretic Field (CHEF) followed by imaging and clustered DNA damages were measured by using Number Average length analysis. Transformation experiments on human primary fibroblast cells using soft agar colony assay were carried out which were irradiated with Fe ions with or without 2 mM melatonin. In plasmid DNA in solution, melatonin reduced the induction of single- and double-strand breaks. Pretreatment of human 28SC cells for 24 h before irradiation with 2 mM melatonin reduced the level of X-ray induced double-strand breaks by {approx}50%, of abasic clustered damages about 40%, and of Fe ion-induced double-strand breaks (41% reduction) and abasic clusters (34% reduction). It decreased transformation to soft agar growth of human primary cells by a factor of 10, but reduced killing by Fe ions only by 20-40%. Melatonin's effective reduction of radiation-induced critical DNA damages, cell killing, and striking decrease of transformation suggest that it is an excellent candidate as a countermeasure against radiation exposure, including radiation exposure to astronaut crews in space travel.

  1. Expression and proliferation profiles of PKC, JNK and p38MAPK in physiologically stretched human bladder smooth muscle cells

    SciTech Connect (OSTI)

    Wazir, Romel; Luo, De-Yi; Dai, Yi; Yue, Xuan; Tian, Ye; Wang, Kun-Jie, E-mail: kunjiewangatscu@163.com

    2013-08-30

    Highlights: •Stretch induces proliferation in human bladder smooth muscle cells (HBSMC). •5% Equibiaxial elongation produces maximum proliferation. •Physiologic stretch decreases apoptotic cell death. •PKC is involved in functional modulation of bladder. •JNK and p38 are not involved in proliferating HBSMC. -- Abstract: Objective: To determine protein kinase C (PKC), c-Jun NH2-Terminal Kinase (JNK) and P38 mitogen-activated protein kinases (p38MAPK) expression levels and effects of their respective inhibitors on proliferation of human bladder smooth muscle cells (HBSMCs) when physiologically stretched in vitro. Materials and methods: HBSMCs were grown on silicone membrane and stretch was applied under varying conditions; (equibiaxial elongation: 2.5%, 5%, 10%, 15%, 20%, 25%), (frequency: 0.05, 0.1, 0.2, 0.5, 1 Hz). Optimal physiological stretch was established by assessing proliferation with 5-Bromo-2-deoxyuridine (BrdU) assay and flow cytometry. PKC, JNK and p38 expression levels were analyzed by Western blot. Specificity was maintained by employing specific inhibitors; (GF109203X for PKC, SP600125 for JNK and SB203580 for p38MAPK), in some experiments. Results: Optimum proliferation was observed at 5% equibiaxial stretch (BrdU: 0.837 ± 0.026 (control) to 1.462 ± 0.023)%, (P < 0.05) and apoptotic cell death rate decreased from 16.4 ± 0.21% (control) to 4.5 ± 0.13% (P < 0.05) applied at 0.1 Hz. Expression of PKC was upregulated with slight increase in JNK and no change in p38MAPK after application of stretch. Inhibition had effects on proliferation (1.075 ± 0.024, P < 0.05 GF109203X); (1.418 ± 0.021, P > 0.05 SP600125) and (1.461 ± 0.01, P > 0.05 SB203580). These findings show that mechanical stretch can promote magnitude-dependent proliferative modulation through PKC and possibly JNK but not via p38MAPK in hBSMCs.

  2. Macrophage-stimulating protein attenuates gentamicin-induced inflammation and apoptosis in human renal proximal tubular epithelial cells

    SciTech Connect (OSTI)

    Lee, Ko Eun [Department of Internal Medicine, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of)] [Department of Internal Medicine, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of); Kim, Eun Young [Department of Physiology, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of)] [Department of Physiology, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of); Kim, Chang Seong; Choi, Joon Seok; Bae, Eun Hui; Ma, Seong Kwon [Department of Internal Medicine, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of)] [Department of Internal Medicine, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of); Kim, Kyung Keun [Department of Pharmacology, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of)] [Department of Pharmacology, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of); Lee, Jong Un [Department of Physiology, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of)] [Department of Physiology, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of); Kim, Soo Wan, E-mail: skimw@chonnam.ac.kr [Department of Internal Medicine, Chonnam National University Medical School, Gwangju 501-757 (Korea, Republic of)

    2013-05-10

    Highlights: •MSP/RON system is activated in rat kidney damaged by gentamicin. •MSP inhibits GM-induced cellular apoptosis and inflammation in HK-2 cells. •MSP attenuates GM-induced activation of MAPKs and NF-?B pathways in HK-2 cells. -- Abstract: The present study aimed to investigate whether macrophage-stimulating protein (MSP) treatment attenuates renal apoptosis and inflammation in gentamicin (GM)-induced tubule injury and its underlying molecular mechanisms. To examine changes in MSP and its receptor, recepteur d’origine nantais (RON) in GM-induced nephropathy, rats were injected with GM for 7 days. Human renal proximal tubular epithelial (HK-2) cells were incubated with GM for 24 h in the presence of different concentrations of MSP and cell viability was measured by MTT assay. Apoptosis was determined by flow cytometry of cells stained with fluorescein isothiocyanate-conjugated annexin V protein and propidium iodide. Expression of Bcl-2, Bax, caspase-3, cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-?B), I?B-?, and mitogen-activated protein kinases (MAPKs) was analyzed by semiquantitative immunoblotting. MSP and RON expression was significantly greater in GM-treated rats, than in untreated controls. GM-treatment reduced HK-2 cell viability, an effect that was counteracted by MSP. Flow cytometry and DAPI staining revealed GM-induced apoptosis was prevented by MSP. GM reduced expression of anti-apoptotic protein Bcl-2 and induced expression of Bax and cleaved caspase 3; these effects and GM-induced expression of COX-2 and iNOS were also attenuated by MSP. GM caused MSP-reversible induction of phospho-ERK, phospho-JNK, and phospho-p38. GM induced NF-?B activation and degradation of I?B-?; the increase in nuclear NF-?B was blocked by inhibitors of ERK, JNK, p-38, or MSP pretreatment. These findings suggest that MSP attenuates GM-induced inflammation and apoptosis by inhibition of the MAPKs/NF-?B signaling pathways.

  3. SIRT1 inactivation induces inflammation through the dysregulation of autophagy in human THP-1 cells

    SciTech Connect (OSTI)

    Takeda-Watanabe, Ai; Kitada, Munehiro; Kanasaki, Keizo; Koya, Daisuke

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer SIRT1 inactivation decreases autophagy in THP-1 cell. Black-Right-Pointing-Pointer Inhibition of autophagy induces inflammation. Black-Right-Pointing-Pointer SIRT1 inactivation induces inflammation through NF-{kappa}B activation. Black-Right-Pointing-Pointer The p62/Sqstm1 accumulation by impairment of autophagy is related to NF-{kappa}B activation. Black-Right-Pointing-Pointer SIRT1 inactivation is involved in the activation of mTOR and decreased AMPK activation. -- Abstract: Inflammation plays a crucial role in atherosclerosis. Monocytes/macrophages are some of the cells involved in the inflammatory process in atherogenesis. Autophagy exerts a protective effect against cellular stresses like inflammation, and it is regulated by nutrient-sensing pathways. The nutrient-sensing pathway includes SIRT1, a NAD{sup +}-dependent histone deacetylase, which is implicated in the regulation of a variety of cellular processes including inflammation and autophagy. The mechanism through which the dysfunction of SIRT1 contributes to the regulation of inflammation in relation to autophagy in monocytes/macrophages is unclear. In the present study, we demonstrate that treatment with 2-[(2-Hydroxynaphthalen-1-ylmethylene)amino]-N-(1-phenethyl)benzamide (Sirtinol), a chemical inhibitor of SIRT1, induces the overexpression of inflammation-related genes such as tumor necrosis factor (TNF)-{alpha} and interleukin (IL)-6 through nuclear factor (NF)-{kappa}B signaling activation, which is associated with autophagy dysfunction, as shown through p62/Sqstm1 accumulation and decreased expression of light chain (LC) 3 II in THP-1 cells. The autophagy inhibitor, 3-methyladenine, also induces inflammation-related NF-{kappa}B activation. In p62/Sqstm1 knockdown cells, Sirtinol-induced inflammation through NF-{kappa}B activation is blocked. In addition, inhibition of SIRT1 is involved in the activation of the mammalian target of rapamycin (mTOR) pathway and is implicated in decreased 5 Prime -AMP activated kinase (AMPK) activation, leading to the impairment of autophagy. The mTOR inhibitor, rapamycin, abolishes Sirtinol-induced inflammation and NF-{kappa}B activation associated with p62/Sqstm1 accumulation. In summary, SIRT1 inactivation induces inflammation through NF-{kappa}B activation and dysregulates autophagy via nutrient-sensing pathways such as the mTOR and AMPK pathways, in THP-1 cells.

  4. MYC transcriptional functions controlling epidermal stem cell self-renewal and differentiation

    E-Print Network [OSTI]

    Nascimento, Elisabete

    IP-on-Chip Protocol . . . . . . . . . . . . . . . . . . . . . . . . 47 2.2.1 Formaldehyde crosslinking of keratinocytes . . . . . . . . . 47 2.2.2 Pre-blocking and binding of antibodies to magnetic beads . 48 2.2.3 Cell sonication... skin was treated with PPAR? ligands. These treatments increase sebocyte differentiation in vitro and sebum production in humans [Trivedi et al., 2006]. 1.3.3 Interfollicular epidermis The interfollicular epidermis is a stratified squamous epithelium...

  5. Hepatitis C virus E2 protein promotes human hepatoma cell proliferation through the MAPK/ERK signaling pathway via cellular receptors

    SciTech Connect (OSTI)

    Zhao Lanjuan [Department of Microbiology, Second Military Medical University, 800 Xiang-Yin Road, Shanghai 200433 (China); Wang Lu [Department of Microbiology, Second Military Medical University, 800 Xiang-Yin Road, Shanghai 200433 (China); Ren Hao [Department of Microbiology, Second Military Medical University, 800 Xiang-Yin Road, Shanghai 200433 (China); Cao Jie [Department of Microbiology, Second Military Medical University, 800 Xiang-Yin Road, Shanghai 200433 (China); Li Li [Department of Laboratory Diagnosis, Changzheng Hospital, Shanghai 200003 (China); Ke Jinshan [Department of Laboratory Diagnosis, Changzheng Hospital, Shanghai 200003 (China); Qi Zhongtian [Department of Microbiology, Second Military Medical University, 800 Xiang-Yin Road, Shanghai 200433 (China)]. E-mail: qizt53@hotmail.com

    2005-04-15

    Dysregulation of mitogen-activated protein kinase (MAPK) signaling pathways by various viruses has been shown to be responsible for viral pathogenicity. The molecular mechanism by which hepatitis C virus (HCV) infection caused human liver diseases has been investigated on the basis of abnormal intracellular signal events. Current data are very limited involved in transmembrane signal transduction triggered by HCV E2 protein. Here we explored regulation of the MAPK/extracellular signal-regulated kinase (MAPK/ERK) signaling pathway by E2 expressed in Chinese hamster oval cells. In human hepatoma Huh-7 cells, E2 specifically activated the MAPK/ERK pathway including downstream transcription factor ATF-2 and greatly promoted cell proliferation. CD81 and low density lipoprotein receptor (LDLR) on the cell surface mediated binding of E2 to Huh-7 cells. The MAPK/ERK activation and cell proliferation driven by E2 were suppressed by blockage of CD81 as well as LDLR. Furthermore, pretreatment with an upstream kinase MEK1/2 inhibitor U0126 also impaired the MAPK/ERK activation and cell proliferation induced by E2. Our results suggest that the MAPK/ERK signaling pathway triggered by HCV E2 via its receptors maintains survival and growth of target cells.

  6. Overexpression of the human BCL-2 gene product results in growth enhancement of Epstein-Barr virus-immortalized B cells

    SciTech Connect (OSTI)

    Tsujimoto, Yoshihide (Wistar Institute of Anatomy and Biology, Philadelphia, PA (USA))

    1989-03-01

    The biological activity of the human BCL-2 gene product was analyzed in an Epstein-Barr virus (EBV)-infected human lymphoblastoid B-cell line transfected with BCL-2 sequences driven by the simian virus 40 promoter and enhancer. Overproduction of the BCL-2 protein conferred a selective growth advantage to the EBV-infected B cells as compared with control transfectants in low-serum medium and also after seeding at limiting dilution but did not render the cells tumorigenic in athymic nude mice. This growth enhancement was also seen in cells transfected with the BCL-2 gene with its own promoter juxtaposed to the immunoglobulin heavy chain gene enhancer, which represents the translocated form of the BCL-2 gene observed in follicular lymphomas with the t(14;18) translocation. The growth advantage of EBV-infected B cells overproducing the BCL-2 protein is neither due to the enhanced growth factor production nor due to an enhanced sensitivity of the BCL-2 transfectants to interleukins 1 or 6, although both lymphokines are known to stimulate proliferation of EBV-infected B-cell lines. The growth advantage of EBV-infected B-cell lines. The growth advantage of EBV-infected B cells by overproduction of the BCL-2 protein suggests the direct involvement of the BCL-2 gene product in the pathogenesis of follicular lymphoma.

  7. The candidate tumor suppressor CST6 alters the gene expression profile of human breast carcinoma cells: Down-regulation of the potent mitogenic, motogenic, and angiogenic factor autotaxin

    SciTech Connect (OSTI)

    Song Jin; Jie Chunfa; Polk, Paula; Shridhar, Ravi; Clair, Timothy; Zhang, Jun; Yin, Lijia; Keppler, Daniel . E-mail: dkeppl@lsuhsc.edu

    2006-02-03

    We recently coined CST6 as a novel candidate tumor suppressor gene for breast cancer. CST6 indeed is expressed in the normal human breast epithelium, but little or not at all in breast carcinomas and breast cancer cell lines. Moreover, ectopic expression of CST6 in human breast cancer cells suppressed cell proliferation, migration, invasion, and orthotopic tumor growth. To obtain insights into the molecular mechanism by which CST6 exhibits its pleiotropic effects on tumor cells, we compared global gene expression profiles in mock- and CST6-transfected human MDA-MB-435S cells. Out of 12,625 transcript species, 61 showed altered expression. These included genes for extracellular matrix components, cytokines, kinases, and phosphatases, as well as several key transcription factors. TaqMan PCR assays were used to confirm the microarray data for 7 out of 11 genes. One down-regulated gene product, secreted autotaxin/lyso-phospholipase D, was of particular interest because its down-regulation by CST6 could explain most of CST6's effect on the breast cancer cells. This study thus provides First evidence that CST6 plays a role in the modulation of genes, particularly, genes that are highly relevant to breast cancer progression.

  8. 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.

  9. NRF2 activation is involved in ozonated human serum upregulation of HO-1 in endothelial cells

    SciTech Connect (OSTI)

    Pecorelli, Alessandra [Department of Molecular and Developmental Medicine, University of Siena (Italy); Child Neuropsychiatry Unit, University Hospital, AOUS, Siena (Italy); Bocci, Velio [Department of Physiology, University of Siena (Italy); Acquaviva, Alessandra [Department of Molecular and Developmental Medicine, University of Siena (Italy); Belmonte, Giuseppe [Department of Biomedical Sciences, University of Siena (Italy); Gardi, Concetta [Department of Molecular and Developmental Medicine, University of Siena (Italy); Virgili, Fabio [INRAN, Rome (Italy); Ciccoli, Lucia [Department of Molecular and Developmental Medicine, University of Siena (Italy); Valacchi, Giuseppe, E-mail: giuseppe.valacchi@unife.it [Department of Life Sciences and Biotechnology, University of Ferrara (Italy); Department of Food and Nutrition, Kyung Hee University, Seoul (Korea, Republic of)

    2013-02-15

    During the last decade, it has been shown that the activation of NRF2 and the binding to electrophile-responsive element (EpREs), stimulates the expression of a great number of genes responsible for the synthesis of phase I and phase II proteins, including antioxidants enzymes and heme oxygenase-1 (HO-1). This critical cell response occurs in cardiovascular, degenerative and chronic infective diseases aggravated by a chronic oxidative stress. In our previous reports we have shown that ozonated plasma is able to up-regulate HO-1 expression in endothelial cells. In the present work we investigated a candidate mechanism involved in this process. After treatment with increasing doses of ozonated serum (20, 40 and 80 ?g/mL O{sub 3} per mL of serum), a clear dose dependent activation of NRF2 and the subsequent induction of HO-1 and NAD(P)H quinone oxidoreductase 1(NQO1) was observed. This effect was also present when cells were treated with serum and hydrogen peroxide (H{sub 2}O{sub 2}) or serum and 4-hydroxynonenal (4HNE). Moreover, the treatment with ozonated serum was associated with a dose-dependent activation of extracellular-signal-regulated kinases (ERK1/2) and p38 MAP kinases (p38), not directly involved in NRF2 activation. These data, provide a new insight on the mechanism responsible for the induction of HO-1 expression by ozonated serum in the endothelium, and have a practical importance as an expedient approach to the treatment of patients with both effective orthodox drugs and ozonated autohemotherapy, targeted to the restoration of redox homeostasis. - Highlights: ? Endothelial HO1 is upregulated by ozonated plasma ? This activation is induced by NRF2 and it is ERK independent. ? 4HNE and H{sub 2}O{sub 2} are the main molecules involved in this process. ? Ozonated plasma induced a hormetic effect ? Combination of orthodox medicine and ozonated plasma can be a useful treatment.

  10. Development of a Breast Cancer Stem Cell Model and the Inhibitory Regulation of Small Molecule Phytochemicals on Various Stages of Human Breast Cancer Cells

    E-Print Network [OSTI]

    Tin, Antony Shen

    2013-01-01

    stem   cells   in   breast   cancer   and   metastasis.  Breast  cancer  research  and  treatment  118(2):241-­?Birnbaum  D  (2009)  Breast  cancer  stem  cells:  tools  

  11. How do microbial fuel cells (MFCs) work? Bacteria need energy to survive, in the same way that humans need food to

    E-Print Network [OSTI]

    Lee, Dongwon

    How do microbial fuel cells (MFCs) work? Bacteria need energy to survive, in the same way that humans need food to live. Bacteria get this energy in a two-step process. The first step requires are receiving more attention because they are a potential part of the solution to our energy demands and could

  12. Genipin-Cross-Linked Microencapsulated Human Adipose Stem Cells Augment Transplant Retention Resulting in Attenuation of Chronically Infarcted Rat Heart Fibrosis and Cardiac Dysfunction

    E-Print Network [OSTI]

    Paul, Arghya; Chen, Guangyong; Khan, Afshan; Rao, Vijayaraghava T. S.; Shum-Tim, Dominique; Prakash, Satya

    2012-12-01

    of biocompatible, fluorogenic genipin-cross-linked alginate chitosan (GCAC) microcapsules in delivery of human adipose stem cells (hASCs) with an aim to increase the implant retention in the infarcted myocardium for maximum clinical benefits. In vitro results show...

  13. First Evaluation of the Biologic Effectiveness Factors of Boron Neutron Capture Therapy (BNCT) in a Human Colon Carcinoma Cell Line

    SciTech Connect (OSTI)

    Dagrosa, Maria Alejandra; Crivello, Martin; Perona, Marina; Thorp, Silvia; Santa Cruz, Gustavo Alberto; Pozzi, Emiliano; Casal, Mariana; Thomasz, Lisa; Cabrini, Romulo; Kahl, Steven; Juvenal, Guillermo Juan; Pisarev, Mario Alberto

    2011-01-01

    Purpose: DNA lesions produced by boron neutron capture therapy (BNCT) and those produced by gamma radiation in a colon carcinoma cell line were analyzed. We have also derived the relative biologic effectiveness factor (RBE) of the neutron beam of the RA-3- Argentine nuclear reactor, and the compound biologic effectiveness (CBE) values for p-boronophenylalanine ({sup 10}BPA) and for 2,4-bis ({alpha},{beta}-dihydroxyethyl)-deutero-porphyrin IX ({sup 10}BOPP). Methods and Materials: Exponentially growing human colon carcinoma cells (ARO81-1) were distributed into the following groups: (1) BPA (10 ppm {sup 10}B) + neutrons, (2) BOPP (10 ppm {sup 10}B) + neutrons, (3) neutrons alone, and (4) gamma rays ({sup 60}Co source at 1 Gy/min dose-rate). Different irradiation times were used to obtain total absorbed doses between 0.3 and 5 Gy ({+-}10%) (thermal neutrons flux = 7.5 10{sup 9} n/cm{sup 2} sec). Results: The frequency of micronucleated binucleated cells and the number of micronuclei per micronucleated binucleated cells showed a dose-dependent increase until approximately 2 Gy. The response to gamma rays was significantly lower than the response to the other treatments (p < 0.05). The irradiations with neutrons alone and neutrons + BOPP showed curves that did not differ significantly from, and showed less DNA damage than, irradiation with neutrons + BPA. A decrease in the surviving fraction measured by 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium bromide (MTT) assay as a function of the absorbed dose was observed for all the treatments. The RBE and CBE factors calculated from cytokinesis block micronucleus (CBMN) and MTT assays were, respectively, the following: beam RBE: 4.4 {+-} 1.1 and 2.4 {+-} 0.6; CBE for BOPP: 8.0 {+-} 2.2 and 2.0 {+-} 1; CBE for BPA: 19.6 {+-} 3.7 and 3.5 {+-} 1.3. Conclusions: BNCT and gamma irradiations showed different genotoxic patterns. To our knowledge, these values represent the first experimental ones obtained for the RA-3 in a biologic model and could be useful for future experimental studies for the application of BNCT to colon carcinoma.

  14. Nitrative DNA damage induced by multi-walled carbon nanotube via endocytosis in human lung epithelial cells

    SciTech Connect (OSTI)

    Guo, Feiye; Ma, Ning; Horibe, Yoshiteru; Kawanishi, Shosuke; Murata, Mariko; Hiraku, Yusuke

    2012-04-15

    Carbon nanotube (CNT) has a promising usage in the field of material science for industrial purposes because of its unique physicochemical property. However, intraperitoneal administration of CNT was reported to cause mesothelioma in experimental animals. Chronic inflammation may contribute to carcinogenesis induced by fibrous materials. 8-Nitroguanine is a mutagenic DNA lesion formed during inflammation and may play a role in CNT-induced carcinogenesis. In this study, we examined 8-nitroguanine formation in A549 human lung alveolar epithelial cells treated with multi-walled CNT (MWCNT) by fluorescent immunocytochemistry. Both MWCNTs with diameter of 20–30 nm (CNT20) and 40–70 nm (CNT40) significantly induced 8-nitroguanine formation at 5 and 10 ?g/ml (p < 0.05), which persisted for 24 h, although there was no significant difference in DNA-damaging abilities of these MWCNTs. MWCNTs significantly induced the expression of inducible nitric oxide synthase (iNOS) for 24 h (p < 0.05). MWCNTs also significantly increased the level of nitrite, a hydrolysis product of oxidized NO, in the culture supernatant at 4 and 8 h (p < 0.05). MWCNT-induced 8-nitroguanine formation and iNOS expression were largely suppressed by inhibitors of iNOS (1400 W), nuclear factor-?B (Bay11-7082), actin polymerization (cytochalasin D), caveolae-mediated endocytosis (methyl-?-cyclodextrin, MBCD) and clathrin-mediated endocytosis (monodansylcadaverine, MDC). Electron microscopy revealed that MWCNT was mainly located in vesicular structures in the cytoplasm, and its cellular internalization was reduced by MBCD and MDC. These results suggest that MWCNT is internalized into cells via clathrin- and caveolae-mediated endocytosis, leading to inflammatory reactions including iNOS expression and resulting nitrative DNA damage, which may contribute to carcinogenesis. Highlights: ?Multi-walled carbon nanotube (MWCNT) caused DNA damage in A549 cells. ?MWCNT formed 8-nitroguanine, a DNA lesion associated with inflammatory response. ?MWCNT was internalized into cells via caveolin- and clathrin-mediated endocytosis. ?8-Nitroguanine formation and iNOS expression involved these types of endocytosis. ?Internalized MWCNT plays a key role in inflammatory response and DNA damage.

  15. Immortalization of normal human mammary epithelial cells in two steps by direct targeting of senescence barriers does not require gross genomic alterations

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

    Garbe, James C.; Vrba, Lukas; Sputova, Klara; Fuchs, Laura; Novak, Petr; Brothman, Arthur R.; Jackson, Mark; Chin, Koei; LaBarge, Mark A.; Watts, George; et al

    2014-10-29

    Telomerase reactivation and immortalization are critical for human carcinoma progression. However, little is known about the mechanisms controlling this crucial step, due in part to the paucity of experimentally tractable model systems that can examine human epithelial cell immortalization as it might occur in vivo. We achieved efficient non-clonal immortalization of normal human mammary epithelial cells (HMEC) by directly targeting the 2 main senescence barriers encountered by cultured HMEC. The stress-associated stasis barrier was bypassed using shRNA to p16INK4; replicative senescence due to critically shortened telomeres was bypassed in post-stasis HMEC by c-MYC transduction. Thus, 2 pathologically relevant oncogenic agentsmore »are sufficient to immortally transform normal HMEC. The resultant non-clonal immortalized lines exhibited normal karyotypes. Most human carcinomas contain genomically unstable cells, with widespread instability first observed in vivo in pre-malignant stages; in vitro, instability is seen as finite cells with critically shortened telomeres approach replicative senescence. Our results support our hypotheses that: (1) telomere-dysfunction induced genomic instability in pre-malignant finite cells may generate the errors required for telomerase reactivation and immortalization, as well as many additional “passenger” errors carried forward into resulting carcinomas; (2) genomic instability during cancer progression is needed to generate errors that overcome tumor suppressive barriers, but not required per se; bypassing the senescence barriers by direct targeting eliminated a need for genomic errors to generate immortalization. Achieving efficient HMEC immortalization, in the absence of “passenger” genomic errors, should facilitate examination of telomerase regulation during human carcinoma progression, and exploration of agents that could prevent immortalization.« less

  16. Immortalization of normal human mammary epithelial cells in two steps by direct targeting of senescence barriers does not require gross genomic alterations

    SciTech Connect (OSTI)

    Garbe, James C.; Vrba, Lukas; Sputova, Klara; Fuchs, Laura; Novak, Petr; Brothman, Arthur R.; Jackson, Mark; Chin, Koei; LaBarge, Mark A.; Watts, George; Futscher, Bernard W.; Stampfer, Martha R.

    2014-10-29

    Telomerase reactivation and immortalization are critical for human carcinoma progression. However, little is known about the mechanisms controlling this crucial step, due in part to the paucity of experimentally tractable model systems that can examine human epithelial cell immortalization as it might occur in vivo. We achieved efficient non-clonal immortalization of normal human mammary epithelial cells (HMEC) by directly targeting the 2 main senescence barriers encountered by cultured HMEC. The stress-associated stasis barrier was bypassed using shRNA to p16INK4; replicative senescence due to critically shortened telomeres was bypassed in post-stasis HMEC by c-MYC transduction. Thus, 2 pathologically relevant oncogenic agents are sufficient to immortally transform normal HMEC. The resultant non-clonal immortalized lines exhibited normal karyotypes. Most human carcinomas contain genomically unstable cells, with widespread instability first observed in vivo in pre-malignant stages; in vitro, instability is seen as finite cells with critically shortened telomeres approach replicative senescence. Our results support our hypotheses that: (1) telomere-dysfunction induced genomic instability in pre-malignant finite cells may generate the errors required for telomerase reactivation and immortalization, as well as many additional “passenger” errors carried forward into resulting carcinomas; (2) genomic instability during cancer progression is needed to generate errors that overcome tumor suppressive barriers, but not required per se; bypassing the senescence barriers by direct targeting eliminated a need for genomic errors to generate immortalization. Achieving efficient HMEC immortalization, in the absence of “passenger” genomic errors, should facilitate examination of telomerase regulation during human carcinoma progression, and exploration of agents that could prevent immortalization.

  17. Bio-inspired nanocomposite assemblies as smart skin components...

    Office of Scientific and Technical Information (OSTI)

    Bio-inspired nanocomposite assemblies as smart skin components. Citation Details In-Document Search Title: Bio-inspired nanocomposite assemblies as smart skin components. There is...

  18. mir-30d Regulates multiple genes in the autophagy pathway and impairs autophagy process in human cancer cells

    SciTech Connect (OSTI)

    Yang, Xiaojun; Department of General Surgery, Gansu Provincial Hospital, Lanzhou, Gansu 710000 ; Zhong, Xiaomin; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011 ; Tanyi, Janos L.; Shen, Jianfeng; Xu, Congjian; Gao, Peng; Zheng, Tim M.; DeMichele, Angela; Zhang, Lin

    2013-02-15

    Highlights: ? Gene set enrichment analysis indicated mir-30d might regulate the autophagy pathway. ? mir-30d represses the expression of BECN1, BNIP3L, ATG12, ATG5 and ATG2. ? BECN1, BNIP3L, ATG12, ATG5 and ATG2 are direct targets of mir-30d. ? mir-30d inhibits autophagosome formation and LC3B-I conversion to LC3B-II. ? mir-30d regulates the autophagy process. -- Abstract: In human epithelial cancers, the microRNA (miRNA) mir-30d is amplified with high frequency and serves as a critical oncomir by regulating metastasis, apoptosis, proliferation, and differentiation. Autophagy, a degradation pathway for long-lived protein and organelles, regulates the survival and death of many cell types. Increasing evidence suggests that autophagy plays an important function in epithelial tumor initiation and progression. Using a combined bioinformatics approach, gene set enrichment analysis, and miRNA target prediction, we found that mir-30d might regulate multiple genes in the autophagy pathway including BECN1, BNIP3L, ATG12, ATG5, and ATG2. Our further functional experiments demonstrated that the expression of these core proteins in the autophagy pathway was directly suppressed by mir-30d in cancer cells. Finally, we showed that mir-30d regulated the autophagy process by inhibiting autophagosome formation and LC3B-I conversion to LC3B-II. Taken together, our results provide evidence that the oncomir mir-30d impairs the autophagy process by targeting multiple genes in the autophagy pathway. This result will contribute to understanding the molecular mechanism of mir-30d in tumorigenesis and developing novel cancer therapy strategy.

  19. Curcumin promotes apoptosis in A549/DDP multidrug-resistant human lung adenocarcinoma cells through an miRNA signaling pathway

    SciTech Connect (OSTI)

    Zhang, Jian, E-mail: zhangjian197011@yahoo.com [Department of Respiratory Medicine, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032 (China)] [Department of Respiratory Medicine, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032 (China); Zhang, Tao [Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an 710038 (China)] [Department of Thoracic Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an 710038 (China); Ti, Xinyu; Shi, Jieran; Wu, Changgui; Ren, Xinling [Department of Respiratory Medicine, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032 (China)] [Department of Respiratory Medicine, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032 (China); Yin, Hong, E-mail: yinnhong@yahoo.com [The Medical Image Center, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032 (China)] [The Medical Image Center, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032 (China)

    2010-08-13

    Research highlights: {yields} Curcumin had anti-cancer effects on A549/DDP multidrug-resistant human lung adenocarcinoma cells {yields} Curcumin promotes apoptosis in A549/DDP cells through a miRNA signaling pathway {yields} Curcumin induces A549/DDP cell apoptosis by downregulating miR-186* {yields} miR-186* may serve as a potential gene therapy target for refractory lung cancer that is sensitive to curcumin -- Abstract: Curcumin extracted from the rhizomes of Curcuma longa L. has been shown to have inhibitory effects on cancers through its anti-proliferative and pro-apoptotic activities. Emerging evidence demonstrates that curcumin can overcome drug resistance to classical chemotherapies. Thus, the mechanisms underlying the anti-tumor activities of curcumin require further study. In our study, we first demonstrated that curcumin had anti-cancer effects on A549/DDP multidrug-resistant human lung adenocarcinoma cells. Further studies showed that curcumin altered miRNA expression; in particular, significantly downregulated the expression of miR-186* in A549/DDP. In addition, transfection of cells with a miR-186* inhibitor promoted A549/DDP apoptosis, and overexpression of miR-186* significantly inhibited curcumin-induced apoptosis in A549/DDP cells. These observations suggest that miR-186* may serve as a potential gene therapy target for refractory lung cancer that is sensitive to curcumin.

  20. Multiple human papillomavirus-16 associated digital squamous-cell carcinomas in an immunocompetent woman with prior human papillomavirus-related genital carcinoma

    E-Print Network [OSTI]

    2011-01-01

    papillomavirus in cutaneous oncogenesis. Ann Plast Surgto contribute to the oncogenesis of anogenital squamous-cell35) are known to contribute to oncogenesis in anogenital and

  1. Skin thickness effects on in vivo LXRF

    SciTech Connect (OSTI)

    Preiss, I.L.; Washington, W. II

    1995-12-31

    The analysis of lead concentration in bone utilizing LXRF can be adversely effected by overlying issue. A quantitative measure of the attenuation of the 10.5 keV Pb L a x-ray signal by skin and skin equivalent plastic has been conducted. Concentration ranges in plaster of Paris and goat bone from 7 to 90 ppm with attenuators of Lucite{reg_sign} and pig skin were examined. It is concluded that no quantitative or semi quantitative analysis can be achieved if overlying sue thickness exceeds 3 mm for Ph concentrations of less than 30 porn Ph in bone.

  2. Hypo-responsiveness of interleukin-8 production in human embryonic epithelial intestine 407 cells independent of NF-{kappa}B pathway: New lessons from endotoxin and ribotoxic deoxynivalenol

    SciTech Connect (OSTI)

    Moon, Yuseok [Department of Microbiology and Immunology, Medical Research Institute, Pusan National University School of Medicine, Busan, 602-739 (Korea, Republic of)], E-mail: moon@pusan.ac.kr; Yang, Hyun; Park, Seung-Hwan [Department of Microbiology and Immunology, Medical Research Institute, Pusan National University School of Medicine, Busan, 602-739 (Korea, Republic of)

    2008-08-15

    Mucosal epithelium senses external toxic insults and transmits the danger signals into the epithelial cells in order to activate a broad range of inflammatory responses. However, pre-exposure to the commensal endotoxins can induce inflammatory tolerance and maintain the homeostasis without excessive immune responses. We recently reported that ribotoxin deoxynivalenol (DON) and its derivatives elicited the pro-inflammatory response as the mucosal insults in human epithelial cells. Taking the knowledge into consideration, we tested the hypothesis that endotoxin pre-exposure can attenuate ribotoxin-induced epithelial interleukin-8 (IL-8) production via a tolerance mechanism. Pre-exposure to endotoxin repressed IL-8 release and its gene expression. However, inflammatory tolerance was not mediated by the attenuated NF-{kappa}B activation which has been generally recognized as the major mediator of LPS-mediated toll-like receptor (TLR) signaling pathway. Instead, pre-exposure to endotoxin was observed to trigger the delayed induction of peroxisome proliferator-activated receptor gamma (PPAR-{gamma}) which contributed to the diminished IL-8 production in the human epithelial cells. Moreover, endogenous PPAR-{gamma} agonist suppressed toxicant-mediated interleukin-8 production and IL-8 mRNA stability. Taken together, endotoxin induced hypo-production of pro-inflammatory cytokine IL-8 in the human epithelial cells, which was associated with the delayed activation of PPAR-{gamma} expression by pre-existing endotoxin.

  3. 'Epidermal Electronics' Paste Peelable Circuitry On Your Skin, Just Like A Temporary Tattoo | Popular Science http://www.popsci.com/science/article/2011-08/epidermal-electronics-paste-peelable-circuitry-your-skin-just-temporary-tattoo[8/14/2011 5:57:38 AM

    E-Print Network [OSTI]

    Rogers, John A.

    - Powered Upgrade New Electric Skin Could Bring the Human Touch to Robots, Artificial Limbs Optical Sensors adhere to the skin not with glue or static electricity, but close-contact atomic forces called van der capacitors and wireless antennas, according to UI. The devices can draw power from induction or even from

  4. Ionizing radiation predisposes non-malignant human mammary epithelial cells to undergo TGF beta-induced epithelial to mesenchymal transition

    E-Print Network [OSTI]

    2007-01-01

    Health Effects; the Low Dose Radiation Program of the DOElong-term, low-dose ionizing radiation exposure in humans.

  5. The skin's role in human thermoregulation and comfort

    E-Print Network [OSTI]

    Arens, Edward A; Zhang, H.

    2006-01-01

    with sensory organs in the hypothalamus (within the brain),within the anterior hypothalamus sense the core temperatureit. The anterior hypothalamus’s warm sensors outnumber its

  6. Athletic equipment microbiota are shaped by interactions with human skin

    Office of Scientific and Technical Information (OSTI)

    AFDC Printable Version Share this resource Send a link to EERE: Alternative Fuels Data Center Home Page to someone by E-mail Share EERE: Alternative Fuels Data Center Home Page on Facebook Tweet about EERE: Alternative Fuels Data Center Home Page on Twitter Bookmark EERE: Alternative Fuels Data Center Homesum_a_epg0_fpd_mmcf_m.xls" ,"Available from WebQuantity of NaturalDukeWakefield MunicipalTechnical Report:Speeding accessby a contractor ofvarDOE PAGES11 PPPL- 4811

  7. Athletic equipment microbiota are shaped by interactions with human skin

    Office of Scientific and Technical Information (OSTI)

    AFDC Printable Version Share this resource Send a link to EERE: Alternative Fuels Data Center Home Page to someone by E-mail Share EERE: Alternative Fuels Data Center Home Page on Facebook Tweet about EERE: Alternative Fuels Data Center Home Page on Twitter Bookmark EERE: Alternative Fuels Data Center Homesum_a_epg0_fpd_mmcf_m.xls" ,"Available from WebQuantity of NaturalDukeWakefield MunicipalTechnical Report:Speeding accessby a contractor ofvarDOE PAGES11 PPPL- 4811(Journal

  8. Athletic equipment microbiota are shaped by interactions with human skin

    Office of Scientific and Technical Information (OSTI)

    AFDC Printable Version Share this resource Send a link to EERE: Alternative Fuels Data Center Home Page to someone by E-mail Share EERE: Alternative Fuels Data Center Home Page on Facebook Tweet about EERE: Alternative Fuels Data Center Home Page on Twitter Bookmark EERE: Alternative Fuels Data Center Homesum_a_epg0_fpd_mmcf_m.xls" ,"Available from WebQuantity of NaturalDukeWakefield MunicipalTechnical Report:Speeding accessby a contractor ofvarDOE PAGES11 PPPL-

  9. Athletic equipment microbiota are shaped by interactions with human skin

    Office of Scientific and Technical Information (OSTI)

    AFDC Printable Version Share this resource Send a link to EERE: Alternative Fuels Data Center Home Page to someone by E-mail Share EERE: Alternative Fuels Data Center Home Page on Facebook Tweet about EERE: Alternative Fuels Data Center Home Page on Twitter Bookmark EERE: Alternative Fuels Data Center Homesum_a_epg0_fpd_mmcf_m.xls" ,"Available from WebQuantity of NaturalDukeWakefieldSulfate Reducing Bacteria (Technical Report) | SciTech Connect Assessing theopacityArticle)Wood et

  10. Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Constructs

    SciTech Connect (OSTI)

    Michael Cornforth

    2012-03-26

    Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. Specific aims apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. The project includes research complementary to NASA/HRP space radiation project.

  11. Regulation of tumor necrosis factor-alpha induced apoptosis via posttranslational modifications in a human colon adenocarcinoma cell line

    E-Print Network [OSTI]

    Kim, Ji-Eun, 1974-

    2004-01-01

    (cont.) phosphoproteomics technology, IMAC/LC/MS/MS, [approximately] 200 phosphosites were identified from HT-29 cells, some of which were detected only from insulin-treated cells. Our phosphoproteomics approach also enabled ...

  12. The effect of temperature on the bystander effect as examined in human prostate carcinoma cells with alpha particle irradiation

    E-Print Network [OSTI]

    Sheppard, Sarah (Sarah Elizabeth)

    2006-01-01

    The bystander effect is seen when irradiated cells release a factor that can produce damage or death in neighboring "bystander" cells that are not actually hit by any radiation. One proposed mechanism involves the irradiated ...

  13. Studying the protein expression in human B lymphoblastoid cells exposed to 1.8-GHz (GSM) radiofrequency radiation (RFR) with protein microarray

    SciTech Connect (OSTI)

    Zhijian, Chen [Department of Environmental and Occupational Health, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang (China) [Department of Environmental and Occupational Health, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, Zhejiang (China); Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China); Xiaoxue, Li [Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China)] [Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China); Wei, Zheng [Zhejiang International Travel Healthcare Center, 230 Zhonghezhong Road, Hangzhou 310003 (China)] [Zhejiang International Travel Healthcare Center, 230 Zhonghezhong Road, Hangzhou 310003 (China); Yezhen, Lu; Jianlin, Lou; Deqiang, Lu; Shijie, Chen; Lifen, Jin [Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China)] [Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China); Jiliang, He, E-mail: he_jiliang@hotmail.com [Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China)] [Institute of Environmental Health, Medical College, Zhejiang University, Hangzhou 310058, Zhejiang (China)

    2013-03-29

    Highlights: ? Protein microarray shows the differential expression of 27 proteins induced by RFR. ? RPA32 related to DNA repair is down-regulated in Western blot. ? p73 related to cell genome stability and apoptosis is up-regulated in Western blot. -- Abstract: In the present study, the protein microarray was used to investigate the protein expression in human B-cell lymphoblastoid cells intermittently exposed to 1.8-GHz GSM radiofrequency radiation (RFR) at the specific absorption rate (SAR) of 2.0 W/kg for 24 h. The differential expression of 27 proteins was found, which were related to DNA damage repair, apoptosis, oncogenesis, cell cycle and proliferation (ratio >1.5-fold, P < 0.05). The results validated with Western blot assay indicated that the expression of RPA32 was significantly down-regulated (P < 0.05) while the expression of p73 was significantly up-regulated in RFR exposure group (P < 0.05). Because of the crucial roles of those proteins in DNA repair and cell apoptosis, the results of present investigation may explain the biological effects of RFR on DNA damage/repair and cell apoptosis.

  14. Inhibitor of Ovarian Cancer Cells Growth by Virtual Screening: A New Thiazole Derivative Targeting Human Thymidylate Synthase

    E-Print Network [OSTI]

    Stroud, Robert

    Inhibitor of Ovarian Cancer Cells Growth by Virtual Screening: A New Thiazole Derivative Targeting-type inhibition pattern, with a Ki of 1.3 M and activity against ovarian cancer cell lines with the same potency ovarian cancer cells. INTRODUCTION The only biosynthetic pathway for 2-deoxythymidine 5- monophosphate

  15. Elevated extracellular calcium increases expression of bone morphogenetic protein-2 gene via a calcium channel and ERK pathway in human dental pulp cells

    SciTech Connect (OSTI)

    Tada, Hiroyuki [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)] [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Nemoto, Eiji, E-mail: e-nemoto@umin.ac.jp [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)] [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Kanaya, Sousuke; Hamaji, Nozomu; Sato, Hisae; Shimauchi, Hidetoshi [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)] [Division of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)

    2010-04-16

    Dental pulp cells, which have been shown to share phenotypical features with osteoblasts, are capable of differentiating into odontoblast-like cells and generating a dentin-like mineral structure. Elevated extracellular Ca{sup 2+}Ca{sub o}{sup 2+} has been implicated in osteogenesis by stimulating the proliferation and differentiation of osteoblasts; however, the role of Ca{sub o}{sup 2+} signaling in odontogenesis remains unclear. We found that elevated Ca{sub o}{sup 2+} increases bone morphogenetic protein (BMP)-2 gene expression in human dental pulp cells. The increase was modulated not only at a transcriptional level but also at a post-transcriptional level, because treatment with Ca{sup 2+} increased the stability of BMP-2 mRNA in the presence of actinomycin D, an inhibitor of transcription. A similar increase in BMP-2 mRNA level was observed in other human mesenchymal cells from oral tissue; periodontal ligament cells and gingival fibroblasts. However, the latter cells exhibited considerably lower expression of BMP-2 mRNA compared with dental pulp cells and periodontal ligament cells. The BMP-2 increase was markedly inhibited by pretreatment with an extracellular signal-regulated kinase (ERK) inhibitor, PD98059, and partially inhibited by the L-type Ca{sup 2+} channels inhibitor, nifedipine. However, pretreatment with nifedipine had no effect on ERK1/2 phosphorylation triggered by Ca{sup 2+}, suggesting that the Ca{sup 2+} influx from Ca{sup 2+} channels may operate independently of ERK signaling. Dental pulp cells do not express the transcript of Ca{sup 2+}-sensing receptors (CaSR) and only respond slightly to other cations such as Sr{sup 2+} and spermine, suggesting that dental pulp cells respond to Ca{sub o}{sup 2+} to increase BMP-2 mRNA expression in a manner different from CaSR and rather specific for Ca{sub o}{sup 2+} among cations.

  16. Dissecting Biological Dark Matter: Single Cell Genetic Analysis of TM7, a Rare and Uncultivated Microbe from the Human Mouth

    E-Print Network [OSTI]

    Marcy, Yann

    2008-01-01

    a Rare and Uncultivated Microbe from the Human Mouth Yannmorphology. Several isolated microbes had a 16S ribosomalmicrobial diversity; microbes colonize a wide variety of

  17. Protein kinase D1 stimulates proliferation and enhances tumorigenesis of MCF-7 human breast cancer cells through a MEK/ERK-dependent signaling pathway

    SciTech Connect (OSTI)

    Karam, Manale; Legay, Christine; Auclair, Christian; Ricort, Jean-Marc, E-mail: jean-marc.ricort@univ-montp2.fr

    2012-03-10

    Protein kinase D1, PKD1, is a novel serine/threonine kinase whose altered expression and dysregulation in many tumors as well as its activation by several mitogens suggest that this protein could regulate proliferation and tumorigenesis. Nevertheless, the precise signaling pathways used are still unclear and the potential direct role of PKD1 in tumor development and progression has not been yet investigated. In order to clarify the role of PKD1 in cell proliferation and tumorigenesis, we studied the effects of PKD1 overexpression in a human adenocarcinoma breast cancer cell line, MCF-7 cells. We demonstrated that overexpression of PKD1 specifically promotes MCF-7 cell proliferation through accelerating G0/G1 to S phase transition of the cell cycle. Moreover, inhibition of endogenous PKD1 significantly reduced cell proliferation. Taken together, these results clearly strengthen the regulatory role of PKD1 in cell growth. We also demonstrated that overexpression of PKD1 specifically diminished serum- and anchorage-dependence for proliferation and survival in vitro and allowed MCF-7 cells to form tumors in vivo. Thus, all these data highlight the central role of PKD1 in biological processes which are hallmarks of malignant transformation. Analysis of two major signaling pathways implicated in MCF-7 cell proliferation showed that PKD1 overexpression significantly increased ERK1/2 phosphorylation state without affecting Akt phosphorylation. Moreover, PKD1 overexpression-stimulated cell proliferation and anchorage-independent growth were totally impaired by inhibition of the MEK/ERK kinase cascade. However, neither of these effects was affected by blocking the PI 3-kinase/Akt signaling pathway. Thus, the MEK/ERK signaling appears to be a determining pathway mediating the biological effects of PKD1 in MCF-7 cells. Taken together, all these data demonstrate that PKD1 overexpression increases the aggressiveness of MCF-7 breast cancer cells through enhancing their oncogenic properties and would, therefore, define PKD1 as a potentially new promising anti-tumor therapeutic target.

  18. MiR-145 is downregulated in human ovarian cancer and modulates cell growth and invasion by targeting p70S6K1 and MUC1

    SciTech Connect (OSTI)

    Wu, Huijuan; Xiao, ZhengHua; Wang, Ke; Liu, Wenxin; Hao, Quan

    2013-11-29

    Highlights: •MiR-145 is downregulated in human ovarian cancer. •MiR-145 targets p70S6K1 and MUC1. •p70S6K1 and MUC1 are involved in miR-145 mediated tumor cell growth and cell invasion, respectively. -- Abstract: MicroRNAs (miRNAs) are a family of small non-coding RNA molecules that regulate gene expression at post-transcriptional levels. Previous studies have shown that miR-145 is downregulated in human ovarian cancer; however, the roles of miR-145 in ovarian cancer growth and invasion have not been fully demonstrated. In the present study, Northern blot and qRT-PCR analysis indicate that miR-145 is downregulated in ovarian cancer tissues and cell lines, as well as in serum samples of ovarian cancer, compared to healthy ovarian tissues, cell lines and serum samples. Functional studies suggest that miR-145 overexpression leads to the inhibition of colony formation, cell proliferation, cell growth viability and invasion, and the induction of cell apoptosis. In accordance with the effect of miR-145 on cell growth, miR-145 suppresses tumor growth in vivo. MiR-145 is found to negatively regulate P70S6K1 and MUC1 protein levels by directly targeting their 3?UTRs. Importantly, the overexpression of p70S6K1 and MUC1 can restore the cell colony formation and invasion abilities that are reduced by miR-145, respectively. MiR-145 expression is increased after 5-aza-CdR treatment, and 5-aza-CdR treatment results in the same phenotype as the effect of miR-145 overexpression. Our study suggests that miR-145 modulates ovarian cancer growth and invasion by suppressing p70S6K1 and MUC1, functioning as a tumor suppressor. Moreover, our data imply that miR-145 has potential as a miRNA-based therapeutic target for ovarian cancer.

  19. 7,12-Dimethylbenzanthracene induces apoptosis in RL95-2 human endometrial cancer cells: Ligand-selective activation of cytochrome P450 1B1

    SciTech Connect (OSTI)

    Kim, Ji Young; Medical Research Science Center, Dong-A University, Busan 602-714 ; Lee, Seung Gee; Mitochondria Hub Regulation Center, Dong-A University, Busan 602-714 ; Chung, Jin-Yong; Medical Research Science Center, Dong-A University, Busan 602-714 ; Kim, Yoon-Jae; Mitochondria Hub Regulation Center, Dong-A University, Busan 602-714 ; Park, Ji-Eun; Medical Research Science Center, Dong-A University, Busan 602-714 ; Oh, Seunghoon; Lee, Se Yong; Choi, Hong Jo; Yoo, Young Hyun; and others

    2012-04-15

    7,12-Dimethylbenzanthracene (DMBA), a polycyclic aromatic hydrocarbon, exhibits mutagenic, carcinogenic, immunosuppressive, and apoptogenic properties in various cell types. To achieve these functions effectively, DMBA is modified to its active form by cytochrome P450 1 (CYP1). Exposure to DMBA causes cytotoxicity-mediated apoptosis in bone marrow B cells and ovarian cells. Although uterine endometrium constitutively expresses CYP1A1 and CYP1B1, their apoptotic role after exposure to DMBA remains to be elucidated. Therefore, we chose RL95-2 endometrial cancer cells as a model system for studying DMBA-induced cytotoxicity and cell death and hypothesized that exposure to DMBA causes apoptosis in this cell type following CYP1A1 and/or CYP1B1 activation. We showed that DMBA-induced apoptosis in RL95-2 cells is associated with activation of caspases. In addition, mitochondrial changes, including decrease in mitochondrial potential and release of mitochondrial cytochrome c into the cytosol, support the hypothesis that a mitochondrial pathway is involved in DMBA-induced apoptosis. Exposure to DMBA upregulated the expression of AhR, Arnt, CYP1A1, and CYP1B1 significantly; this may be necessary for the conversion of DMBA to DMBA-3,4-diol-1,2-epoxide (DMBA-DE). Although both CYP1A1 and CYP1B1 were significantly upregulated by DMBA, only CYP1B1 exhibited activity. Moreover, knockdown of CYP1B1 abolished DMBA-induced apoptosis in RL95-2 cells. Our data show that RL95-2 cells are susceptible to apoptosis by exposure to DMBA and that CYP1B1 plays a pivotal role in DMBA-induced apoptosis in this system. -- Highlights: ? Cytotoxicity-mediated apoptogenic action of DMBA in human endometrial cancer cells. ? Mitochondrial pathway in DMBA-induced apoptosis of RL95-2 endometrial cancer cells. ? Requirement of ligand-selective activation of CYP1B1 in DMBA-induced apoptosis.

  20. Raman spectroscopy of single human tumour cells exposed to ionizing radiation in vitro This article has been downloaded from IOPscience. Please scroll down to see the full text article.

    E-Print Network [OSTI]

    Brolo, Alexandre G.

    Raman spectroscopy of single human tumour cells exposed to ionizing radiation in vitro This article tissue samples which will survive long enough in a lab to perform a radiation experiment. Animal in non-human systems to radiation therapy patient outcomes. As such, prescribed doses for tumour control

  1. Nonstochastic effects of different energy beta emitters on pig skin

    SciTech Connect (OSTI)

    Peel, D.M.; Hopewell, J.W.; Wells, J.; Charles, M.W.

    1984-08-01

    Circular areas of pig skin from 1- to 40-mm diameter were irradiated with ..beta.. emitters of high, medium, and low energies, /sup 90/Sr, /sup 170/Tm, and /sup 147/Pm, respectively. The study provides information for radiological protection problems of localized skin exposures. During the first 16 weeks after irradiation /sup 90/Sr produced a first reaction due to epithelial cell death followed by a second reaction attributable to damage to the dermal blood vessels. /sup 170/Tm and /sup 147/Pm produced the epithelial reaction only. The epithelial dose response varied as a function of ..beta.. energy. The doses required to produce moist desquamation in 50% of 15- to 22.5-mm fields (ED/sub 50/) were 30-45 Gy from/sup 90/Sr, approx.80 Gy from /sup 170/Tm, and approx.500 Gy from /sup 147/Pm. An area effect was observed in the epithelial response to /sup 90/Sr irradiation. The ED/sub 50/ for moist desquamation ranged from approx.25 Gy for a 40-mm source to approx.450 Gy for a 1-mm source. It is also suggested that the area effects could be explained by different modes of epithelial repopulation after irradiation.

  2. Engineering humanized mice for improved hematopoietic reconstitution

    E-Print Network [OSTI]

    Drake, Adam

    Humanized mice are immunodeficient animals engrafted with human hematopoietic stem cells that give rise to various lineages of human blood cells throughout the life of the mouse. This article reviews recent advances in the ...

  3. The accumulations of HIF-1? and HIF-2? by JNK and ERK are involved in biphasic effects induced by different levels of arsenite in human bronchial epithelial cells

    SciTech Connect (OSTI)

    Xu, Yuan; Li, Yuan [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China) [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); Li, Huiqiao [Qujing Center for Disease Control and Prevention, Qujing 655000, Yunnan (China)] [Qujing Center for Disease Control and Prevention, Qujing 655000, Yunnan (China); Pang, Ying; Zhao, Yue; Jiang, Rongrong; Shen, Lu [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China) [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); Zhou, Jianwei; Wang, Xinru [The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China)] [The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); Liu, Qizhan, E-mail: drqzliu@hotmail.com [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China) [Institute of Toxicology, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China); The Key Laboratory of Modern Toxicology, Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 210029, Jiangsu (China)

    2013-01-15

    The biphasic effects of arsenite, in which low levels of arsenite induce cell proliferation and high levels of arsenite induce DNA damage and apoptosis, apparently contribute to arsenite-induced carcinogenesis. However, the mechanisms underlying this phenomenon are not well understood. In this study, we investigated the effects of different levels of arsenite on cell proliferation, DNA damage and apoptosis as well as on signal transduction pathways in human bronchial epithelial (HBE) cells. Our results show that a low level of arsenite activates extracellular signal-regulated kinases (ERK), which probably mediate arsenite-inhibited degradation of ubiquitinated hypoxia-inducible factor-2? (HIF-2?) in HBE cells. ERK inhibition blocks cell proliferation induced by a low level of arsenite, in part via HIF-2?. In contrast, a high level of arsenite activates c-Jun N-terminal kinases (JNK), which provoke a response to suppress ubiquitinated HIF-1? degradation. Down-regulation of HIF-1? by inhibiting JNK, however, increases the DNA damage but decreases the apoptosis induced by a high level of arsenite. Thus, data in the present study suggest that the accumulations of HIF-1? and HIF-2? by JNK and ERK are involved in different levels of arsenite-induced biphasic effects, with low levels of arsenite inducing cell proliferation and high levels of arsenite inducing DNA damage and apoptosis in HBE cells. -- Highlights: ? Biphasic effects induced by different concentrations of arsenite. ? Different regulation of ERK or JNK signal pathway by arsenite. ? Different regulation of HIF1? or HIF 2? by arsenite.

  4. 4-Hydroxyestradiol induces oxidative stress and apoptosis in human mammary epithelial cells: possible protection by NF-{kappa}B and ERK/MAPK

    SciTech Connect (OSTI)

    Chen Zhihua [National Research Laboratory of Molecular Carcinogenesis and Chemoprevention, College of Pharmacy, Seoul National University, Shinlim-dong, Kwanak-ku, Seoul 151-742 (Korea, Republic of); Na, Hye-Kyung [National Research Laboratory of Molecular Carcinogenesis and Chemoprevention, College of Pharmacy, Seoul National University, Shinlim-dong, Kwanak-ku, Seoul 151-742 (Korea, Republic of); Hurh, Yeon-Jin [National Research Laboratory of Molecular Carcinogenesis and Chemoprevention, College of Pharmacy, Seoul National University, Shinlim-dong, Kwanak-ku, Seoul 151-742 (Korea, Republic of); Surh, Young-Joon [National Research Laboratory of Molecular Carcinogenesis and Chemoprevention, College of Pharmacy, Seoul National University, Shinlim-dong, Kwanak-ku, Seoul 151-742 (Korea, Republic of)]. E-mail: surh@plaza.snu.ac.kr

    2005-10-01

    Catechol estrogens, the hydroxylated metabolites of 17{beta}-estradiol (E{sub 2}), have been considered to be implicated in estrogen-induced carcinogenesis. 4-Hydroxyestradiol (4-OHE{sub 2}), an oxidized metabolite of E{sub 2} formed preferentially by cytochrome P450 1B1, reacts with DNA to form depurinating adducts thereby exerting genotoxicity and carcinogenicity. 4-OHE{sub 2} undergoes 2-electron oxidation to quinone via semiquinone, and during this process, reactive oxygen species (ROS) can be generated to cause DNA damage and cell death. In the present study, 4-OHE{sub 2} was found to elicit cytotoxicity in cultured human mammary epithelial (MCF-10A) cells, which was blocked by the antioxidant trolox. MCF-10A cells treated with 4-OHE{sub 2} exhibited increased intracellular ROS accumulation and 8-oxo-7,8-dihydroxy-2'-deoxyguanosine formation, and underwent apoptosis as determined by poly(ADP-ribose)polymerase cleavage and disruption of mitochondrial transmembrane potential. The redox-sensitive transcription factor nuclear factor {kappa}B (NF-{kappa}B) was transiently activated by 4-OHE{sub 2} treatment. Cotreatment of MCF-10A cells with the NF-{kappa}B inhibitor, L-1-tosylamido-2-phenylethyl chloromethyl ketone, exacerbated 4-OHE{sub 2}-induced cell death. 4-OHE{sub 2} also caused transient activation of extracellular signal-regulated protein kinases (ERK) involved in transmitting cell survival or death signals. A pharmacological inhibitor of ERK aggravated the 4-OHE{sub 2}-induced cytotoxicity, supporting the pivotal role of ERK in protecting against catechol estrogen-induced oxidative cell death.

  5. A polysaccharide fraction of adlay seed (Coixlachryma-jobi L.) induces apoptosis in human non-small cell lung cancer A549 cells

    SciTech Connect (OSTI)

    Lu, Xiangyi; Liu, Wei; Wu, Junhua; Li, Mengxian [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China)] [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China); Wang, Juncheng; Wu, Jihui [School of Life Science, University of Science and Technology of China, Hefei 230022 (China)] [School of Life Science, University of Science and Technology of China, Hefei 230022 (China); Luo, Cheng, E-mail: Luo58@yahoo.com [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China)] [Key Laboratory of Food Nutrition and Safety, Ministry of Education, School of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457 (China)

    2013-01-11

    Highlights: Black-Right-Pointing-Pointer A polysaccharide from adlay seed, its molecular mass, optical rotation and sugars was determined. Black-Right-Pointing-Pointer We demonstrated that a polysaccharide from adlay can induce apoptosis in cancer cells. Black-Right-Pointing-Pointer The polysaccharide inhibited the metabolism and proliferation of NSCLC A549 cells. Black-Right-Pointing-Pointer The polysaccharide may trigger apoptosis via the mitochondria-dependent pathway. -- Abstract: Different seed extracts from Coix lachryma-jobi (adlay seed) have been used for the treatment of various cancers in China, and clinical data support the use of these extracts for cancer therapy; however, their underlying molecular mechanisms have not been well defined. A polysaccharide fraction, designated as CP-1, was extracted from the C.lachryma-jobi L. var. using the ethanol subsiding method. CP-1 induced apoptosis in A549 cells in a dose-dependent manner, as determined by MTT assay. Apoptotic bodies were observed in the cells by scanning electronic microscopy. Apoptosis and DNA accumulation during S-phase of the cell cycle were determined by annexin V-FITC and PI staining, respectively, and measured by flow cytometry. CP-1 also extended the comet tail length on single cell gel electrophoresis, and disrupted the mitochondrial membrane potential. Further analysis by western blotting showed that the expression of caspase-3 and caspase-9 proteins was increased. Taken together, our results demonstrate that CP-1 is capable of inhibiting A549 cell proliferation and inducing apoptosis via a mechanism primarily involving the activation of the intrinsic mitochondrial pathway. The assay data suggest that in addition to its nutritional properties, CP-1 is a very promising candidate polysaccharide for the development of anti-cancer medicines.

  6. The effects of Shiga toxin 1 on cytokine and chemokine production and apoptosis in a human monocytic cell line 

    E-Print Network [OSTI]

    Harrison, Lisa Margaret

    2004-11-15

    receptor, Gb3, on endothelial cell surfaces. A main source of proinflammatory cytokines is the macrophage, thus leading us to utilize the monocytic/macrophage-like cell line, THP-1, as a model for cytokine production in Stx pathogenesis. In addition...

  7. Ran GTPase protein promotes human pancreatic cancer proliferation by deregulating the expression of Survivin and cell cycle proteins

    SciTech Connect (OSTI)

    Deng, Lin; Department of Oncology, Tangdu Hospital, Fourth Military Medical University, Xi’an, Shaanxi 710038 ; Lu, Yuanyuan; Zhao, Xiaodi; Sun, Yi; Shi, Yongquan; Fan, Hongwei; Liu, Changhao; Zhou, Jinfeng; Nie, Yongzhan; Wu, Kaichun; Fan, Daiming; Guo, Xuegang

    2013-10-18

    Highlights: •Overexpression of Ran in pancreatic cancer was correlated with histological grade. •Downregulation of Ran could induce cell apoptosis and inhibit cell proliferation. •The effects were mediated by cell cycle proteins, Survivin and cleaved Caspase-3. -- Abstract: Ran, a member of the Ras GTPase family, has important roles in nucleocytoplasmic transport. Herein, we detected Ran expression in pancreatic cancer and explored its potential role on tumour progression. Overexpressed Ran in pancreatic cancer tissues was found highly correlated with the histological grade. Downregulation of Ran led to significant suppression of cell proliferation, cell cycle arrest at the G1/S phase and induction of apoptosis. In vivo studies also validated that result. Further studies revealed that those effects were at least partly mediated by the downregulation of Cyclin A, Cyclin D1, Cyclin E, CDK2, CDK4, phospho-Rb and Survivin proteins and up regulation of cleaved Caspase-3.

  8. Dynamic Skin Triangulation Ho-Lun Cheng

    E-Print Network [OSTI]

    Edelsbrunner, Herbert

    that grow and shrink with time. An example is the boundary between the solid and the liquid portions the nu- cleation, growth and coarsening stages [1]. Moving bound- aries also arise naturally in mold the finite collection by convex combina- tion and shrinking. The skin surface is the envelope of this family

  9. Dynamic Skin Triangulation Ho-Lun Cheng

    E-Print Network [OSTI]

    Sullivan, John M.

    of physical simulation, where they act as bound- aries of spatial domains that grow and shrink with time stages [1]. Moving bound- aries also arise naturally in mold filling processes, both for metal and other and shrinking. The skin surface is the envelope of this family. Even though the family is infinite, the surface

  10. Isolation and effects of citrus limonoids on cytochrome p450 inhibition, apoptotic induction and cytotoxicity on human cancer cells

    E-Print Network [OSTI]

    Poulose, Shibu M.

    2007-04-25

    This dissertation illustrates an efficient purification method for citrus limonoids and flavonoids, while examining their effects on cytochrome P450 inhibition and apoptotic induction on human neuroblastoma (SH-SY5Y) ...

  11. Effects of oncogenic Ras and p38 mitogen-activated protein kinase on the adhesion of normal human cells

    E-Print Network [OSTI]

    Waldman, Lynne K

    2010-01-01

    Activating mutations in RAS oncogenes commonly arise in human cancers. However, in experimental settings, oncogenic RAS has most often been studied at supraphysiological levels of expression. Importantly, work by others ...

  12. Mechanisms of hormonal regulation of CAD gene expression and inhibition by Aryl hydrocarbon receptor agonist in human breast cancer cells 

    E-Print Network [OSTI]

    Khan, Shaheen Munawar Ali

    2007-04-25

    The CAD gene is trifunctional and expresses carbamoylphosphate synthetase/aspartate carbamyltransferase/dihydroorotase, which are required for pyrimidine biosynthesis. CAD gene activities are induced in MCF-7 human breast ...

  13. Antibacterial agent triclosan suppresses RBL-2H3 mast cell function

    SciTech Connect (OSTI)

    Palmer, Rachel K.; Hutchinson, Lee M.; Burpee, Benjamin T.; Tupper, Emily J.; Pelletier, Jonathan H.; Kormendy, Zsolt; Hopke, Alex R.; Malay, Ethan T.; Evans, Brieana L.; Velez, Alejandro; Gosse, Julie A.

    2012-01-01

    Triclosan is a broad-spectrum antibacterial agent, which has been shown previously to alleviate human allergic skin disease. The purpose of this study was to investigate the hypothesis that the mechanism of this action of triclosan is, in part, due to effects on mast cell function. Mast cells play important roles in allergy, asthma, parasite defense, and carcinogenesis. In response to various stimuli, mast cells degranulate, releasing allergic mediators such as histamine. In order to investigate the potential anti-inflammatory effect of triclosan on mast cells, we monitored the level of degranulation in a mast cell model, rat basophilic leukemia cells, clone 2H3. Having functional homology to human mast cells, as well as a very well defined signaling pathway leading to degranulation, this cell line has been widely used to gain insight into mast-cell driven allergic disorders in humans. Using a fluorescent microplate assay, we determined that triclosan strongly dampened the release of granules from activated rat mast cells starting at 2 ?M treatment, with dose-responsive suppression through 30 ?M. These concentrations were found to be non-cytotoxic. The inhibition was found to persist when early signaling events (such as IgE receptor aggregation and tyrosine phosphorylation) were bypassed by using calcium ionophore stimulation, indicating that the target for triclosan in this pathway is likely downstream of the calcium signaling event. Triclosan also strongly suppressed F-actin remodeling and cell membrane ruffling, a physiological process that accompanies degranulation. Our finding that triclosan inhibits mast cell function may explain the clinical data mentioned above and supports the use of triclosan or a mechanistically similar compound as a topical treatment for allergic skin disease, such as eczema. -- Highlights: ?The effects of triclosan on mast cell function using a murine mast cell model. ?Triclosan strongly inhibits degranulation of mast cells. ?Triclosan suppresses membrane ruffling of activated mast cells. ?Triclosan's effects persist when early mast cell signaling events are bypassed. ?Supports use of triclosan as a topical treatment for eczema.

  14. Cell Motility and Deformability in the Pathogenesis of Lung Cancer

    E-Print Network [OSTI]

    Pagano, Paul Carmelo

    2015-01-01

    might prevent a cell from moving at full capacity because itfull malignant phenotype on human bronchial epithelial cells.full malignant phenotype on human bronchial epithelial cells.

  15. Hypoxia-inducible factor-1? (HIF-1?) is upregulated in a HIF-1?-dependent manner in 518A2 human melanoma cells under hypoxic conditions

    SciTech Connect (OSTI)

    Mandl, Markus, E-mail: mmandl@mail.austria.com; Kapeller, Barbara; Lieber, Roman; Macfelda, Karin

    2013-04-26

    Highlights: •HIF-1? is a hypoxia-responsive protein in 518A2 human melanoma cells. •HIF-1? is upregulated in a HIF-1?-dependent manner under hypoxic conditions. •HIF-1? is not elevated due to heterodimerization with HIF-1? per se. •HIF-1? inducibility has a biological relevance as judged in Het-CAM model. -- Abstract: Solid tumors include hypoxic areas due to excessive cell proliferation. Adaptation to low oxygen levels is mediated by the hypoxia-inducible factor (HIF) pathway promoting invasion, metastasis, metabolic alterations, chemo-resistance and angiogenesis. The transcription factor HIF-1, the major player within this pathway consists of HIF-1? and HIF-1?. The alpha subunit is continuously degraded under normoxia and becomes stabilized under reduced oxygen supply. In contrast, HIF-1? is generally regarded as constitutively expressed and being present in excess within the cell. However, there is evidence that the expression of this subunit is more complex. The aim of this study was to investigate the role of HIF-1? in human melanoma cells. Among a panel of five different cell lines, in 518A2 cells exposed to the hypoxia-mimetic cobalt chloride HIF-1? was rapidly elevated on protein level. Knockdown experiments performed under cobalt chloride-exposure and hypoxia revealed that this effect was mediated by HIF-1?. The non-canonical relationship between these subunits was further confirmed by pharmacologic inhibition of HIF-1? and by expression of a dominant-negative HIF mutant. Overexpression of HIF-1? showed a time delay in HIF-1? induction, thus arguing for HIF-1? de novo synthesis rather than protein stabilization by heterodimerization. A Hen’s egg test-chorioallantoic membrane model of angiogenesis and invasion indicated a local expression of HIF-1? and implies a biological relevance of these findings. In summary, this study demonstrates the HIF-1?-dependent regulation of HIF-1? under hypoxic conditions for the first time. The results indicate a novel cell specific mechanism which might prevent HIF-1? to become a limiting factor.

  16. Chemical Agent Induced Reduction of Skin Light Scattering 

    E-Print Network [OSTI]

    Hirshburg, Jason M.

    2011-02-22

    Skin turbidity limits light based medical applications while increasing the risk of epidermal thermal injury. Collagen fibers are responsible for the majority of light scattering within skin. Chemicals, known as clearing ...

  17. Vaccine delivery with microneedle skin patches in nonhuman primates

    E-Print Network [OSTI]

    Li, Adrienne V

    Transcutaneous drug delivery from planar skin patches is effective for small-molecule drugs and skin-permeable vaccine adjuvants. However, to achieve efficient delivery of vaccines and other macromolecular therapeutics ...

  18. Dietary chromium and nickel enhance UV-carcinogenesis in skin...

    Office of Scientific and Technical Information (OSTI)

    chromium and nickel enhance UV-carcinogenesis in skin of hairless mice The skin cancer enhancing effect of chromium (in male mice) and nickel in UVR-irradiated female Skh1...

  19. Osthole inhibits the invasive ability of human lung adenocarcinoma cells via suppression of NF-?B-mediated matrix metalloproteinase-9 expression

    SciTech Connect (OSTI)

    Kao, Shang-Jyh [Department of Chest Medicine, Shin-Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan (China) [Department of Chest Medicine, Shin-Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan (China); School of Respiratory Therapy, Taipei Medical University, Taipei Taiwan (China); Su, Jen-Liang [Graduate Institute of Cancer Biology, College of Medicine, China Medical University, Taichung, Taiwan (China) [Graduate Institute of Cancer Biology, College of Medicine, China Medical University, Taichung, Taiwan (China); Center for Molecular Medicine, China Medical University Hospital, Taichung, Taiwan (China); Department of Biotechnology, Asia University, Taichung, Taiwan (China); Chen, Chi-Kuan [Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan (China)] [Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan (China); Yu, Ming-Chih; Bai, Kuan-Jen; Chang, Jer-Hua [Division of Pulmonary Medicine, Department of Internal Medicine, Taipei Medical University-Wan Fang Hospital, Taipei, Taiwan (China)] [Division of Pulmonary Medicine, Department of Internal Medicine, Taipei Medical University-Wan Fang Hospital, Taipei, Taiwan (China); Bien, Mauo-Ying [School of Respiratory Therapy, Taipei Medical University, Taipei Taiwan (China) [School of Respiratory Therapy, Taipei Medical University, Taipei Taiwan (China); Division of Pulmonary Medicine, Department of Internal Medicine, Taipei Medical University-Wan Fang Hospital, Taipei, Taiwan (China); Yang, Shun-Fa [Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan (China) [Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan (China); Department of Medical Research, Chung Shan Medical University Hospital, Taichung, Taiwan (China); Chien, Ming-Hsien, E-mail: mhchien1976@gmail.com [Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan (China)] [Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan (China)

    2012-05-15

    The induction of matrix metalloproteinase (MMP)-9 is particularly important for the invasiveness of various cancer cells. Osthole, a natural coumarin derivative extracted from traditional Chinese medicines, is known to inhibit the proliferation of a variety of tumor cells, but the effect of osthole on the invasiveness of tumor cells is largely unknown. This study determines whether and by what mechanism osthole inhibits invasion in CL1-5 human lung adenocarcinoma cells. Herein, we found that osthole effectively inhibited the migratory and invasive abilities of CL1-5 cells. A zymographic assay showed that osthole inhibited the proteolytic activity of MMP-9 in CL1-5 cells. Inhibition of migration, invasion, and MMP2 and/or MMP-9 proteolytic activities was also observed in other lung adenocarcinoma cell lines (H1299 and A549). We further found that osthole inhibited MMP-9 expression at the messenger RNA and protein levels. Moreover, a chromatin immunoprecipitation assay showed that osthole inhibited the transcriptional activity of MMP-9 by suppressing the DNA binding activity of nuclear factor (NF)-?B in the MMP-9 promoter. Using reporter assays with point-mutated promoter constructs further confirmed that the inhibitory effect of osthole requires an NF-?B binding site on the MMP-9 promoter. Western blot and immunofluorescence assays demonstrated that osthole inhibited NF-?B activity by inhibiting I?B-? degradation and NF-?B p65 nuclear translocation. In conclusion, we demonstrated that osthole inhibits NF-?B-mediated MMP-9 expression, resulting in suppression of lung cancer cell invasion and migration, and osthole might be a potential agent for preventing the invasion and metastasis of lung cancer. -- Highlights: ? Osthole treatment inhibits lung adenocarcinoma cells migration and invasion. ? Osthole reduces the expression and proteolytic activity of MMP-9. ? Osthole inhibits MMP-9 transcription via suppression of NF-?B binding activity. ? Osthole inhibits I?B? degradation and NF-?B nucleus translocation. ? Osthole suppresses EMT by repressing vimentin and inducing E-cadherin expression.

  20. Detection of human hematopoietic stem cell engraftment in the livers of adult immunodeficient mice by an optimized flow cytometric method

    E-Print Network [OSTI]

    Varga, Nicole L.; Bárcena, Alicia; Fomin, Marina E.; Muench, Marcus O.

    2010-01-01

    myeloid cell develop- ment in NOD/ltsz-scid IL2R gamma nullblood and immune systems in NOD/SCID/IL2 receptor {gamma}platelet production assessed in NOD/SCID mice injected with

  1. Generation and characterization of transgene-free human induced pluripotent stem cells and conversion to putative clinical-grade status

    E-Print Network [OSTI]

    2013-01-01

    Cell 2010, 7:618–630. 15. Sommer CA, Stadtfeld M, Murphy GJ,27:543–549. 16. Somers A, Jean JC, Sommer CA, Omari A, FordCC, Mills JA, Ying L, Sommer AG, Jean JM, Smith BW, Lafyatis

  2. DNA Double-Strand Breaks Form in Bystander Cells after Microbeam Irradiation of Three-dimensional Human Tissue Models

    E-Print Network [OSTI]

    Brenner, David Jonathan

    Research Accelerator Facility, Center for Radiological Research, College of Physicians and Surgeons Department of Biological Sciences, University of Lethbridge, Lethbridge, Alberta, Canada; and 3 Radiological implications for cancer radiother- apy and diagnostic radiology as well as for human health in general

  3. Structure, function and diversity of the healthy human microbiome

    E-Print Network [OSTI]

    Alm, Eric J.

    Studies of the human microbiome have revealed that even healthy individuals differ remarkably in the microbes that occupy habitats such as the gut, skin and vagina. Much of this diversity remains unexplained, although diet, ...

  4. Diospyrin derivative, an anticancer quinonoid, regulates apoptosis at endoplasmic reticulum as well as mitochondria by modulating cytosolic calcium in human breast carcinoma cells

    SciTech Connect (OSTI)

    Kumar, Binod [Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032 (India) [Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032 (India); Radiation and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Kumar, Amit [Radiation and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)] [Radiation and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Ghosh, Subhalakshmi [Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032 (India)] [Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032 (India); Pandey, Badri N., E-mail: bnp@barc.gov.in [Radiation and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Mishra, Kaushala P. [Radiation and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)] [Radiation and Cancer Biology Section, Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Hazra, Banasri, E-mail: banasrihazra@yahoo.co.in [Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032 (India)] [Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032 (India)

    2012-01-13

    Highlights: Black-Right-Pointing-Pointer Diospyrin diethylether (D7) caused oxidative stress-dependent activation of PC-PLC. Black-Right-Pointing-Pointer Activated PC-PLC induced a sustained-release of Ca{sup 2+} from endoplasmic reticulum. Black-Right-Pointing-Pointer The elevated cytosolic Ca{sup +2} led to the calpain-caspase12 dependent apoptosis. Black-Right-Pointing-Pointer D7-Induced Ca{sup +2} also found to accentuate the mitochondrial pathway of apoptosis. -- Abstract: Diospyrin diethylether (D7), a bisnaphthoquinonoid derivative, exhibited an oxidative stress-dependent apoptosis in several human cancer cells and tumor models. The present study was aimed at evaluation of the increase in cytosolic calcium [Ca{sup 2+}]{sub c} leading to the apoptotic cell death triggered by D7 in MCF7 human breast carcinoma cells. A phosphotidylcholine-specific phospholipase C (PC-PLC) inhibitor, viz. U73122, and an antioxidant, viz. N-acetylcysteine, could significantly prevent the D7-induced rise in [Ca{sup 2+}]{sub c} and PC-PLC activity. Using an endoplasmic reticulum (ER)-Ca{sup 2+} mobilizer (thapsigargin) and an ER-IP3R antagonist (heparin), results revealed ER as a major source of [Ca{sup 2+}]{sub c} which led to the activation of calpain and caspase12, and cleavage of fodrin. These effects including apoptosis were significantly inhibited by the pretreatment of Bapta-AM (a cell permeable Ca{sup 2+}-specific chelator), or calpeptin (a calpain inhibitor). Furthermore, D7-induced [Ca{sup 2+}]{sub c} was found to alter mitochondrial membrane potential and induce cytochrome c release, which was inhibited by either Bapta-AM or ruthenium red (an inhibitor of mitochondrial Ca{sup 2+} uniporter). Thus, these results provided a deeper insight into the D7-induced redox signaling which eventually integrated the calcium-dependent calpain/caspase12 activation and mitochondrial alterations to accentuate the induction of apoptotic cell death.

  5. Meeting Report. Assessing Human Germ-Cell Mutagenesis in the Post-Genome Era: A Celebration of the Legacy of William Lawson (Bill) Russell

    E-Print Network [OSTI]

    2006-01-01

    the landmark Human Genome Project (HGP) and its relationshipworking on the Human Genome Project and the Online Mendelianthat presaged the Human Genome Project (listed below) and

  6. miR-421 induces cell proliferation and apoptosis resistance in human nasopharyngeal carcinoma via downregulation of FOXO4

    SciTech Connect (OSTI)

    Chen, Liang; Department of Otolaryngology, Guangzhou General Hospital of PLA Guangzhou Command, Guangzhou 510010 ; Tang, Yanping; Wang, Jian; Yan, Zhongjie; Xu, Ruxiang

    2013-06-14

    Highlights: •miR-421 is upregulated in nasopharyngeal carcinoma. •miR-421 induces cell proliferation and apoptosis resistance. •FOXO4 is a direct and functional target of miR-421. -- Abstract: microRNAs have been demonstrated to play important roles in cancer development and progression. Hence, identifying functional microRNAs and better understanding of the underlying molecular mechanisms would provide new clues for the development of targeted cancer therapies. Herein, we reported that a microRNA, miR-421 played an oncogenic role in nasopharyngeal carcinoma. Upregulation of miR-421 induced, whereas inhibition of miR-421 repressed cell proliferation and apoptosis resistance. Furthermore, we found that upregulation of miR-421 inhibited forkhead box protein O4 (FOXO4) signaling pathway following downregulation of p21, p27, Bim and FASL expression by directly targeting FOXO4 3?UTR. Additionally, we demonstrated that FOXO4 expression is critical for miR-421-induced cell growth and apoptosis resistance. Taken together, our findings not only suggest that miR-421 promotes nasopharyngeal carcinoma cell proliferation and anti-apoptosis, but also uncover a novel regulatory mechanism for inactivation of FOXO4 in nasopharyngeal carcinoma.

  7. Lysophosphatidic acid induces reactive oxygen species generation by activating protein kinase C in PC-3 human prostate cancer cells

    SciTech Connect (OSTI)

    Lin, Chu-Cheng; Lin, Chuan-En; Lin, Yueh-Chien [Institute of Zoology, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China)] [Institute of Zoology, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China); Ju, Tsai-Kai [Instrumentation Center, National Taiwan University, Taipei, Taiwan, ROC (China) [Instrumentation Center, National Taiwan University, Taipei, Taiwan, ROC (China); Technology Commons, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China); Huang, Yuan-Li [Department of Biotechnology, Asia University, Taichung, Taiwan, ROC (China)] [Department of Biotechnology, Asia University, Taichung, Taiwan, ROC (China); Lee, Ming-Shyue [Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan, ROC (China)] [Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan, ROC (China); Chen, Jiun-Hong [Institute of Zoology, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China) [Institute of Zoology, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China); Department of Life Science, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China); Lee, Hsinyu, E-mail: hsinyu@ntu.edu.tw [Institute of Zoology, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China) [Institute of Zoology, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China); Department of Life Science, College of Life Science, National Taiwan University, Taipei, Taiwan, ROC (China); Center for Biotechnology, National Taiwan University, Taipei, Taiwan, ROC (China); Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan, ROC (China)

    2013-11-01

    Highlights: •LPA induces ROS generation through LPA{sub 1} and LPA{sub 3}. •LPA induces ROS generation by activating PLC. •PKC? mediates LPA-induced ROS generation. -- Abstract: Prostate cancer is one of the most frequently diagnosed cancers in males, and PC-3 is a cell model popularly used for investigating the behavior of late stage prostate cancer. Lysophosphatidic acid (LPA) is a lysophospholipid that mediates multiple behaviors in cancer cells, such as proliferation, migration and adhesion. We have previously demonstrated that LPA enhances vascular endothelial growth factor (VEGF)-C expression in PC-3 cells by activating the generation of reactive oxygen species (ROS), which is known to be an important mediator in cancer progression. Using flow cytometry, we showed that LPA triggers ROS generation within 10 min and that the generated ROS can be suppressed by pretreatment with the NADPH oxidase (Nox) inhibitor diphenylene iodonium. In addition, transfection with LPA{sub 1} and LPA{sub 3} siRNA efficiently blocked LPA-induced ROS production, suggesting that both receptors are involved in this pathway. Using specific inhibitors and siRNA, phospholipase C (PLC) and protein kinase C (PKC) were also suggested to participate in LPA-induced ROS generation. Overall, we demonstrated that LPA induces ROS generation in PC-3 prostate cancer cells and this is mediated through the PLC/PKC/Nox pathway.

  8. The novel BH-3 mimetic apogossypolone induces Beclin-1- and ROS-mediated autophagy in human hepatocellular carcinoma cells

    E-Print Network [OSTI]

    Cheng, P.; Ni, Zhenhong; Dai, Xufang; Wang, Bin; Ding, W.; Smith, A Rae; Xu, Liang; Wu, D.; He, Fengtian; Lian, Jiqin

    2013-02-07

    , Pervaiz S. Simultaneous induction of non-canonical autophagy and apoptosis in cancer cells by ROS-dependent ERK and JNK activation. PLoS One 2010; 5: e9996. 34. Yuan Y, Xue X, Guo R-B, Sun X-L, Hu G. Resveratrol enhances the antitumor effects...

  9. Estrogen induced concentration dependent differential gene expression in human breast cancer (MCF7) cells: Role of transcription factors

    SciTech Connect (OSTI)

    Chandrasekharan, Sabarinath; Kandasamy, Krishna Kumar; Dayalan, Pavithra; Ramamurthy, Viraragavan

    2013-08-02

    Highlights: •Estradiol (E2) at low dose induced cell proliferation in breast cancer cells. •E2 at high concentration induced cell stress in breast cancer cells. •Estrogen receptor physically interacts only with a few transcription factors. •Differential expression of genes with Oct-1 binding sites increased under stress. •Transcription factor binding sites showed distinct spatial distribution on genes. -- Abstract: Background: Breast cancer cells respond to estrogen in a concentration dependent fashion, resulting in proliferation or apoptosis. The mechanism of this concentration dependent differential outcome is not well understood yet. Methodology: Meta-analysis of the expression data of MCF7 cells treated with low (1 nM) or high (100 nM) dose of estradiol (E2) was performed. We identified genes differentially expressed at the low or the high dose, and examined the nature of regulatory elements in the vicinity of these genes. Specifically, we looked for the difference in the presence, abundance and spatial distribution of binding sites for estrogen receptor (ER) and selected transcription factors (TFs) in the genomic region up to 25 kb upstream and downstream from the transcription start site (TSS) of these genes. Results: It was observed that at high dose E2 induced the expression of stress responsive genes, while at low dose, genes involved in cell cycle were induced. We found that the occurrence of transcription factor binding regions (TFBRs) for certain factors such as Sp1 and SREBP1 were higher on regulatory regions of genes expressed at low dose. At high concentration of E2, genes with a higher frequency of Oct-1 binding regions were predominantly involved. In addition, there were differences in the spatial distribution pattern of the TFBRs in the genomic regions among the two sets of genes. Discussion: E2 induced predominantly proliferative/metabolic response at low concentrations; but at high concentration, stress–rescue responses were induced. At high E2 concentration, classical genomic pathway involving ER binding to the regulatory regions was reduced, and alternate or indirect activation of genes through Oct-1 became more prominent.

  10. Extracellular acidification induces connective tissue growth factor production through proton-sensing receptor OGR1 in human airway smooth muscle cells

    SciTech Connect (OSTI)

    Matsuzaki, Shinichi; Ishizuka, Tamotsu; Yamada, Hidenori; Kamide, Yosuke; Hisada, Takeshi; Ichimonji, Isao; Aoki, Haruka; Yatomi, Masakiyo; Komachi, Mayumi; Tsurumaki, Hiroaki; Ono, Akihiro; Koga, Yasuhiko; Dobashi, Kunio; Mogi, Chihiro; Sato, Koichi; Tomura, Hideaki; Mori, Masatomo; Okajima, Fumikazu

    2011-10-07

    Highlights: {yields} The involvement of extracellular acidification in airway remodeling was investigated. {yields} Extracellular acidification alone induced CTGF production in human ASMCs. {yields} Extracellular acidification enhanced TGF-{beta}-induced CTGF production in human ASMCs. {yields} Proton-sensing receptor OGR1 was involved in acidic pH-stimulated CTGF production. {yields} OGR1 may play an important role in airway remodeling in asthma. -- Abstract: Asthma is characterized by airway inflammation, hyper-responsiveness and remodeling. Extracellular acidification is known to be associated with severe asthma; however, the role of extracellular acidification in airway remodeling remains elusive. In the present study, the effects of acidification on the expression of connective tissue growth factor (CTGF), a critical factor involved in the formation of extracellular matrix proteins and hence airway remodeling, were examined in human airway smooth muscle cells (ASMCs). Acidic pH alone induced a substantial production of CTGF, and enhanced transforming growth factor (TGF)-{beta}-induced CTGF mRNA and protein expression. The extracellular acidic pH-induced effects were inhibited by knockdown of a proton-sensing ovarian cancer G-protein-coupled receptor (OGR1) with its specific small interfering RNA and by addition of the G{sub q/11} protein-specific inhibitor, YM-254890, or the inositol-1,4,5-trisphosphate (IP{sub 3}) receptor antagonist, 2-APB. In conclusion, extracellular acidification induces CTGF production through the OGR1/G{sub q/11} protein and inositol-1,4,5-trisphosphate-induced Ca{sup 2+} mobilization in human ASMCs.

  11. Inhibition of the ERK phosphorylation plays a role in terbinafine-induced p21 up-regulation and DNA synthesis inhibition in human vascular endothelial cells

    SciTech Connect (OSTI)

    Ho, P.-Y. [Graduate Institute of Cell and Molecular Biology, Taipei Medical University, Taipei, Taiwan (China); Hsu, S.-P. [Institute of Physiology, College of Medicine, National Taiwan University, Taiwan (China); Liang, Y.-C. [Graduate Institute of Biomedical Technology, Taipei Medical University, Taipei, Taiwan (China); Kuo, M.-L. [Institute of Toxicology, College of Medicine, National Taiwan University, Taiwan (China); Ho, Y.-S. [Graduate Institute of Biomedical Technology, Taipei Medical University, Taipei, Taiwan (China); Lee, W.-S. [Graduate Institute of Neuroscience, Taipei Medical University, Taipei, Taiwan (China); Graduate Institute of Medical Sciences, Taipei Medical University, 250 Wu-Hsing Street, Taipei 110, Taiwan (China); Department of Physiology, Medical College, Taipei Medical University, Taipei, Taiwan (China)], E-mail: wslee@tmu.edu.tw

    2008-05-15

    Previously, we showed that terbinafine (TB) induces cell-cycle arrest in cultured human umbilical vein endothelial cells (HUVEC) through an up-regulation of the p21 protein. The aim of this study is to delineate the molecular mechanisms underlying TB-induced increase of p21 protein. RT-PCR analysis demonstrated that the mRNA levels of p21 and p53 were increased in the TB-treated HUVEC. The p21 promoter activity was also increased by TB treatment. Transfection of HUVEC with p53 dominant negative (DN) abolished the TB-induced increases of p21 promoter activity and protein level, suggesting that the TB-induced increase of p21 is p53-dependent. Western blot analysis demonstrated that TB decreased the levels of phosphorylated extracellular signal-regulated kinase (ERK). Over-expression of mitogen-activated protein kinase (MEK)-1, the immediate upstream activator kinase of ERK, abolished the TB-induced increases of p21 and p53 protein and decrease of thymidine incorporation. The ERK inhibitor (PD98059) enhanced the TB-induced inhibition of thymidine incorporation into HUVEC. Taken together, these data suggest that the decrease of ERK activity plays a role in the TB-induced up-regulation of p21 in HUVEC. On the other hand, pretreatment of the cells with geranylgeraniol (GGOH), farnesol (FOH), or Ras inhibitor peptide did not affect the TB-induced decrease of thymidine incorporation. Taken together, our results suggest that TB might cause a decrease of MEK, which in turn up-regulates p53 through the inhibition of ERK phosphorylation, and finally causes an increase of p21 expression and cell-cycle arrest.

  12. Telomerase Regulation and Telomere Maintenance during Human T-cell Leukemia/Lymphoma Virus (HTLV-I) Transformation

    E-Print Network [OSTI]

    Bellon, Marcia Lynn

    2008-04-29

    and the Graduate Faculty of the University of Kansas Medical Center in partial fulfillment of the requirements for the degree of Doctor of Philosophy. _____________________________? Chairperson:??? Committee...RNA expression PBMC (gray bar), HTLV-I negative and positive leukemic cell lines in the order shown in A (black bars). Statistical analysis by the student t-tests against PBMC control was performed and values are 42 presented. Increased TRF1, TRF2, POT-1...

  13. Quantitative proteomic analysis of the inhibitory effects of CIL-102 on viability and invasiveness in human glioma cells

    SciTech Connect (OSTI)

    Teng, Chih-Chuan; Kuo, Hsing-Chun; Sze, Chun-I

    2013-11-01

    CIL-102 (1-[4-(furo[2,3-b]quinolin-4-ylamino)phenyl]ethanone), the major active agent of the alkaloid derivative, has been demonstrated to exert anticancer effects. Herein, we present an investigation focused on the identification of the target(s) of CIL-102's action and the mechanism of its action in apoptotic and anti-invasive pathways. Proteomic approaches were used to purify and identify the protein substrates using 2D difference gel electrophoresis (2D SDS-PAGE) to assess changes in the expression of relevant protein treatment with CIL-102 that resulted in the inhibition of viability and invasion. Our results demonstrate that CIL-102 treatment of U87 cells decreased cell proliferation and invasiveness. CIL-102 dose-dependent induction of apoptosis and inhibitory invasiveness were accompanied by sustained phosphorylation of JNK1/2 and p70S6K as well as generation of the reactive oxygen species. In addition, differential proteins displayed between CIL-102-treated and untreated U87 were determined and validated. There were 11 differentially expressed proteins between the CIL-102-treated and untreated groups. Furthermore, we demonstrated that CIL-102 inhibited cancer cell proliferation and reduced anti-invasion properties by up-regulating the levels of FUMH (Fumarate hydratase). The investigation demonstrated that there was an increase in the cellular levels of FUMH in the CIL-102 reduction in viability and invasion via the activation of JNK1/2 and mTOR signaling modules. NAC administration and shRNA FUMH conferred resistance to CIL-102-inhibited HIF1? and MMP-2 levels via inhibition of JNK1/2 and mTOR activation. We concluded that CIL-102-induced an apoptosis cascade and decreased aggressiveness in astrocytoma cells by modulation of mitochondria function, providing a new mechanism for CIL-102 treatment. - Highlights: • We found the effect of CIL-102 on neuroblastoma cells. • Fumarate hydratase as a CIL-102's target by proteomic differential displays. • CIL-102 regulated-FUMH stimulates apoptosis-related protein and inactivation HIF1.

  14. Development of a combined model of tissue kinetics and radiation response of human bronchiolar epithelium with single cell resolution 

    E-Print Network [OSTI]

    Ostrovskaya, Natela Grigoryevna

    2006-10-30

    cells of the airways due to internal exposure to alpha-particle emitters, e.g. radon. Inhalation of radon, a colorless and odorless gas, one of the products of the decay of uranium which occurs naturally in the earth?s crust, is the second major cause... epithelial tissue plays an important role in normal lung physiology. square4 lung epithelia are target tissues for occupational internal exposures and for radon exposure (26); square4 the epithelium of bronchioles appears to be the origin...

  15. Turbine blade having a constant thickness airfoil skin

    DOE Patents [OSTI]

    Marra, John J

    2012-10-23

    A turbine blade is provided for a gas turbine comprising: a support structure comprising a base defining a root of the blade and a framework extending radially outwardly from the base, and an outer skin coupled to the support structure framework. The skin has a generally constant thickness along substantially the entire radial extent thereof. The framework and the skin define an airfoil of the blade.

  16. Parsing ERK Activation Reveals Quantitatively Equivalent Contributions From Epidermal Growth Factor Receptor and HER2 In Human Mammary Epithelial Cells

    SciTech Connect (OSTI)

    Hendriks, Bart S.; Orr, Galya; Wells, Alan H.; Wiley, H. S.; Lauffenburger, Douglas A.

    2005-02-18

    HER2, a member of the EGFR tyrosine kinase family, functions as an accessory EGFR signaling component and alters EGFR trafficking by heterodimerization. HER2 overexpression leads to aberrant cell behavior including enhanced proliferation and motility. Here we apply a combination of computational modeling and quantitative experimental studies of the dynamic interactions between EGFR and HER2, and their downstream activation of extracellular signal-related kinase (ERK) to understand this complex signaling system. Using cells expressing different levels of HER2 relative to the EGFR, we can separate relative contributions of EGFR and HER2 to signaling amplitude and duration. Based on our model calculations, we demonstrate that, in contrast with previous suggestions in the literature, the intrinsic capabilities of EGFR and HER2 to activated ERK are quantitatively equivalent . We find that HER2-mediated effects on EGFR dimerization and trafficking are sufficient to explain the detected HER2-mediated amplification of EGF-induced ERK signaling. Our model suggests that transient amplification of ERK activity by HER2 arises predominantly from the 2-to-1 stoichiometry of receptor kinase to bound ligand in EGFR/HER2 heterodimers compared to the 1-to-1 stoichiometry of the EGFR homodimer, but alterations in receptor trafficking, with resultant EGFR sparing, cause the sustained HER2-mediated enhancement of ERK signaling.

  17. Validation of in vitro cell models used in drug metabolism and transport studies; genotyping of cytochrome P450, phase II enzymes and drug transporter polymorphisms in the human hepatoma (HepG2), ovarian carcinoma (IGROV-1) and colon carcinoma (CaCo-2, LS180) cell lines

    SciTech Connect (OSTI)

    Brandon, Esther F.A.; Bosch, Tessa M.; Deenen, Maarten J.; Levink, Rianne; Wal, Everdina van der; Meerveld, Joyce B.M. van; Bijl, Monique; Beijnen, Jos H. |; Schellens, Jan H.M. |; Meijerman, Irma . E-mail: I.Meijerman@pharm.uu.nl

    2006-02-15

    Human cell lines are often used for in vitro biotransformation and transport studies of drugs. In vivo, genetic polymorphisms have been identified in drug-metabolizing enzymes and ABC-drug transporters leading to altered enzyme activity, or a change in the inducibility of these enzymes. These genetic polymorphisms could also influence the outcome of studies using human cell lines. Therefore, the aim of our study was to pharmacogenotype four cell lines frequently used in drug metabolism and transport studies, HepG2, IGROV-1, CaCo-2 and LS180, for genetic polymorphisms in biotransformation enzymes and drug transporters. The results indicate that, despite the presence of some genetic polymorphisms, no real effects influencing the activity of metabolizing enzymes or drug transporters in the investigated cell lines are expected. However, this characterization will be an aid in the interpretation of the results of biotransformation and transport studies using these in vitro cell models.

  18. The Direction of Optimal Skin Incisions Derived From Striae Distensae.

    E-Print Network [OSTI]

    Lemperle, Gottfried

    2015-01-01

    of Optimal Skin Incisions Derived from Striae Distensaestriae lines. These are derived from the striae compositecal incisions and scars derived from the Internet. Incision

  19. Orai1 Function is Essential for T Cell Homing to Lymph Nodes

    E-Print Network [OSTI]

    Greenberg, Milton L.; Yu, Ying; Leverrier, Sabrina; Zhang, Shenyuan L.; Parker, Ian; Cahalan, Michael D.

    2013-01-01

    Biotech). In some experiments, human CD3 + T cells were7 human PBL were injected intraperitoneally and experimentsindicated. Human cells were used for experiments 24 hr after

  20. Regulation of insulin-like growth factor binding protein-4 in Ah responsive and Ah nonresponsive human breast cancer cells by 17 B-estradiol and 2,3,7,8 tetrachlorodibenzo-p-dioxin 

    E-Print Network [OSTI]

    Schrope, Katherine Eillene

    1997-01-01

    This study focused on the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on insulin-like growth factor binding protein-4 (IGFBP-4) protein secretion and MRNA expression. In human breast cancer cells, 17 O-estradiol (E2) induced IGFBP-4...

  1. TOWARDS AUTOMATED SKIN LESION DIAGNOSIS Paul Wighton

    E-Print Network [OSTI]

    Atkins, M. Stella

    for minimal risk human research, by the Office of Research Ethics. A copy of the approval letter has been be improved. Typically, the ASLD pipeline consists of

  2. Detecting pornographic images by localizing skin Sotiris Karavarsamisa

    E-Print Network [OSTI]

    Blekas, Konstantinos

    specialized sub- classes, namely "bikini" / "porn" and "skin" / "non-skin", respectively. The extracted pornographic image classifiers. Index Terms convex hull calculation, multi-class classification, porn detection unprotected Web services in order to circulate or exchange child pornography and general pornographic content

  3. INVESTIGATION The Lsktm1 Locus Modulates Lung and Skin

    E-Print Network [OSTI]

    Broman, Karl W.

    INVESTIGATION The Lsktm1 Locus Modulates Lung and Skin Tumorigenesis in the Mouse Antonella Galvan to both skin and lung tumorigenesis over the susceptibility of the SWR/J strain. In an effort to map tumor.93) and lung (LOD score = 8.74) tumorigenesis. Two genes, Igfbp5 and Igfbp2, residing in this locus

  4. Neutron skin of 208 Pb in consistency with

    E-Print Network [OSTI]

    Neutron skin of 208 Pb in consistency with neutron star observations K. Miyazaki E-mail: miyazakiro as varying the neutron radius of 208Pb. The neutron skin thickness Sn is determined in the comparison with the astronomical observations of massive neutron stars (NSs), the standard scenario of NS cooling

  5. Skin cancer detection by oblique-incidence diffuse reflectance spectroscopy 

    E-Print Network [OSTI]

    Smith, Elizabeth Brooks

    2009-05-15

    Skin cancer is the most common form of cancer and it is on the rise. If skin cancer is diagnosed early enough, the survival rate is close to 90%. Oblique-incidence diffuse reflectance (OIR) spectroscopy offers a technology that may be used...

  6. Laminin peptide YIGSR induces collagen synthesis in Hs27 human dermal fibroblasts

    SciTech Connect (OSTI)

    Yoon, Jong Hyuk; Kim, Jaeyoon; Lee, Hyeongjoo [NovaCell Technology Inc., Pohang, Kyungbuk 790-784 (Korea, Republic of)] [NovaCell Technology Inc., Pohang, Kyungbuk 790-784 (Korea, Republic of); Kim, So Young [Department of Dermatology, Chung-Ang University College of Medicine, Seoul 156-756 (Korea, Republic of) [Department of Dermatology, Chung-Ang University College of Medicine, Seoul 156-756 (Korea, Republic of); Department of Convergence Medicine and Pharmaceutical Biosciences, Graduate School, Chung-Ang University, Seoul 156-756 (Korea, Republic of); Jang, Hwan-Hee [Functional Food and Nutrition Division, Department of Agrofood Resources, Rural Development Administration, Suwon 441-853 (Korea, Republic of)] [Functional Food and Nutrition Division, Department of Agrofood Resources, Rural Development Administration, Suwon 441-853 (Korea, Republic of); Ryu, Sung Ho [Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Pohang, Kyungbuk 790-784 (Korea, Republic of)] [Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology (POSTECH), Pohang, Kyungbuk 790-784 (Korea, Republic of); Kim, Beom Joon [Department of Dermatology, Chung-Ang University College of Medicine, Seoul 156-756 (Korea, Republic of) [Department of Dermatology, Chung-Ang University College of Medicine, Seoul 156-756 (Korea, Republic of); Department of Convergence Medicine and Pharmaceutical Biosciences, Graduate School, Chung-Ang University, Seoul 156-756 (Korea, Republic of); Lee, Taehoon G., E-mail: taehoon@novacelltech.com [NovaCell Technology Inc., Pohang, Kyungbuk 790-784 (Korea, Republic of)

    2012-11-23

    Highlights: Black-Right-Pointing-Pointer We identify a function of the YIGSR peptide to enhance collagen synthesis in Hs27. Black-Right-Pointing-Pointer YIGSR peptide enhanced collagen type 1 synthesis both of gene and protein levels. Black-Right-Pointing-Pointer There were no changes in cell proliferation and MMP-1 level in YIGSR treatment. Black-Right-Pointing-Pointer The YIGSR effect on collagen synthesis mediated activation of FAK, pyk2 and ERK. Black-Right-Pointing-Pointer The YIGSR-induced FAK and ERK activation was modulated by FAK and MEK inhibitors. -- Abstract: The dermal ECM is synthesized from fibroblasts and is primarily compromised of fibrillar collagen and elastic fibers, which support the mechanical strength and resiliency of skin, respectively. Laminin, a major glycoprotein located in the basement membrane, promotes cell adhesion, cell growth, differentiation, and migration. The laminin tyrosine-isoleucine-glycine-serine-arginine (YIGSR) peptide, corresponding to the 929-933 sequence of the {beta}1 chain, is known to be a functional motif with effects on the inhibition of tumor metastasis, the regulation of sensory axonal response and the inhibition of angiogenesis through high affinity to the 67 kDa laminin receptor. In this study, we identified a novel function of the YIGSR peptide to enhance collagen synthesis in human dermal fibroblasts. To elucidate this novel function regarding collagen synthesis, we treated human dermal fibroblasts with YIGSR peptide in both a time- and dose-dependent manner. According to subsequent experiments, we found that the YIGSR peptide strongly enhanced collagen type 1 synthesis without changing cell proliferation or cellular MMP-1 level. This YIGSR peptide-mediated collagen type 1 synthesis was modulated by FAK inhibitor and MEK inhibitor. This study clearly reveals that YIGSR peptide plays a novel function on the collagen type 1 synthesis of dermal fibroblasts and also suggests that YIGSR is a strong candidate peptide for the treatment of skin aging and wrinkles.

  7. Method and apparatus to measure the depth of skin burns

    DOE Patents [OSTI]

    Dickey, Fred M. (Albuquerque, NM); Holswade, Scott C. (Albuquerque, NM)

    2002-01-01

    A new device for measuring the depth of surface tissue burns based on the rate at which the skin temperature responds to a sudden differential temperature stimulus. This technique can be performed without physical contact with the burned tissue. In one implementation, time-dependent surface temperature data is taken from subsequent frames of a video signal from an infrared-sensitive video camera. When a thermal transient is created, e.g., by turning off a heat lamp directed at the skin surface, the following time-dependent surface temperature data can be used to determine the skin burn depth. Imaging and non-imaging versions of this device can be implemented, thereby enabling laboratory-quality skin burn depth imagers for hospitals as well as hand-held skin burn depth sensors the size of a small pocket flashlight for field use and triage.

  8. Integrated analysis reveals that STAT3 is central to the crosstalk between HER/ErbB receptor signaling pathways in human mammary epithelial cells

    SciTech Connect (OSTI)

    Gong, Chunhong; Zhang, Yi; Shankaran, Harish; Resat, Haluk

    2015-01-01

    Human epidermal growth factor receptors (HER, also known as ErbB) drive cellular proliferation, pro-survival and stress responses by activating several downstream kinases, in particular ERK, p38, JNK (SAPK), the PI3K/AKT, as well as various transcriptional regulators such as STAT3. When co-expressed, first three members of HER family (HER1-3) can form homo- and hetero-dimers. Based on the considerable evidence which suggest that every receptor dimer activates intracellular signaling pathways differentially, we hypothesized that the HER dimerization pattern is a better predictor of downstream signaling than the total receptor activation levels. We validated our hypothesis using a combination of model-based analysis to quantify the HER dimerization patterns and multi-factorial experiments where HER dimerization patterns and signaling crosstalk were rationally perturbed. We have measured the activation of HER1-3 receptors and of the sentinel signaling proteins ERK, AKT, p38, JNK, STAT3 as a function of time in a panel of human mammary epithelial (HME) cells expressing different levels of HER1-3 stimulated with various ligand combinations. Our analysis using multiple ways of clustering the activation data has confirmed that the HER receptor dimer is a better predictor of the signaling through p38, ERK and AKT pathways than the total HER receptor expression and activation levels. Targeted inhibition studies to identify the causal effects allowed us to obtain a consensus regulatory interaction model, which revealed that STAT3 occupies a central role in the crosstalk between the studied pathways.

  9. Fibroblast growth factor 2 inhibits up-regulation of bone morphogenic proteins and their receptors during osteoblastic differentiation of human mesenchymal stem cells

    SciTech Connect (OSTI)

    Biver, Emmanuel, E-mail: ebiver@yahoo.fr [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France) [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France); Department of Rheumatology, Lille University Hospital, Roger Salengro Hospital, 59037 Lille cedex (France); Service of Bone Diseases, Department of Internal Medicine Specialties, University Hospital of Geneva, CH-1211 Geneva 14 (Switzerland); Soubrier, Anne-Sophie [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France) [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France); Department of Rheumatology, Lille University Hospital, Roger Salengro Hospital, 59037 Lille cedex (France); Thouverey, Cyril [Service of Bone Diseases, Department of Internal Medicine Specialties, University Hospital of Geneva, CH-1211 Geneva 14 (Switzerland)] [Service of Bone Diseases, Department of Internal Medicine Specialties, University Hospital of Geneva, CH-1211 Geneva 14 (Switzerland); Cortet, Bernard [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France) [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France); Department of Rheumatology, Lille University Hospital, Roger Salengro Hospital, 59037 Lille cedex (France); Broux, Odile [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France)] [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France); Caverzasio, Joseph [Service of Bone Diseases, Department of Internal Medicine Specialties, University Hospital of Geneva, CH-1211 Geneva 14 (Switzerland)] [Service of Bone Diseases, Department of Internal Medicine Specialties, University Hospital of Geneva, CH-1211 Geneva 14 (Switzerland); Hardouin, Pierre [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France)] [Physiopathology of Inflammatory Bone Diseases, EA 4490, University Lille North of France, Quai Masset, Bassin Napoleon, BP120, 62327 Boulogne sur Mer (France)

    2012-11-02

    Highlights: Black-Right-Pointing-Pointer FGF modulates BMPs pathway in HMSCs by down-regulating BMP/BMPR expression. Black-Right-Pointing-Pointer This effect is mediated by ERK and JNK MAPKs pathways. Black-Right-Pointing-Pointer Crosstalk between FGF and BMPs must be taken into account in skeletal bioengineering. Black-Right-Pointing-Pointer It must also be considered in the use of recombinant BMPs in orthopedic and spine surgeries. -- Abstract: Understanding the interactions between growth factors and bone morphogenic proteins (BMPs) signaling remains a crucial issue to optimize the use of human mesenchymal stem cells (HMSCs) and BMPs in therapeutic perspectives and bone tissue engineering. BMPs are potent inducers of osteoblastic differentiation. They exert their actions via BMP receptors (BMPR), including BMPR1A, BMPR1B and BMPR2. Fibroblast growth factor 2 (FGF2) is expressed by cells of the osteoblastic lineage, increases their proliferation and is secreted during the healing process of fractures or in surgery bone sites. We hypothesized that FGF2 might influence HMSC osteoblastic differentiation by modulating expressions of BMPs and their receptors. BMP2, BMP4, BMPR1A and mainly BMPR1B expressions were up-regulated during this differentiation. FGF2 inhibited HMSCs osteoblastic differentiation and the up-regulation of BMPs and BMPR. This effect was prevented by inhibiting the ERK or JNK mitogen-activated protein kinases which are known to be activated by FGF2. These data provide a mechanism explaining the inhibitory effect of FGF2 on osteoblastic differentiation of HMSCs. These crosstalks between growth and osteogenic factors should be considered in the use of recombinant BMPs in therapeutic purpose of fracture repair or skeletal bioengineering.

  10. Liver X receptor alpha mediated genistein induction of human dehydroepiandrosterone sulfotransferase (hSULT2A1) in Hep G2 cells

    SciTech Connect (OSTI)

    Chen, Yue; Zhang, Shunfen [Department of Physiological Sciences, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078 (United States); Zhou, Tianyan [Department of Pharmaceutics, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100083 (China); Huang, Chaoqun; McLaughlin, Alicia [Department of Physiological Sciences, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078 (United States); Chen, Guangping, E-mail: guangping.chen@okstate.edu [Department of Physiological Sciences, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078 (United States)

    2013-04-15

    Cytosolic sulfotransferases are one of the major families of phase II drug metabolizing enzymes. Sulfotransferase-catalyzed sulfonation regulates hormone activities, metabolizes drugs, detoxifies xenobiotics, and bioactivates carcinogens. Human dehydroepiandrosterone sulfotransferase (hSULT2A1) plays important biological roles by sulfating endogenous hydroxysteroids and exogenous xenobiotics. Genistein, mainly existing in soy food products, is a naturally occurring phytoestrogen with both chemopreventive and chemotherapeutic potential. Our previous studies have shown that genistein significantly induces hSULT2A1 in Hep G2 and Caco-2 cells. In this study, we investigated the roles of liver X receptor (LXR?) in the genistein induction of hSULT2A1. LXRs have been shown to induce expression of mouse Sult2a9 and hSULT2A1 gene. Our results demonstrate that LXR? mediates the genistein induction of hSULT2A1, supported by Western blot analysis results, hSULT2A1 promoter driven luciferase reporter gene assay results, and mRNA interference results. Chromatin immunoprecipitation (ChIP) assay results demonstrate that genistein increase the recruitment of hLXR? binding to the hSULT2A1 promoter. These results suggest that hLXR? plays an important role in the hSULT2A1 gene regulation. The biological functions of phytoestrogens may partially relate to their induction activity toward hydroxysteroid SULT. - Highlights: ? Liver X receptor ? mediated genistein induction of hSULT2A1 in Hep G2 cells. ? LXR? and RXR? dimerization further activated this induction. ? Western blot results agreed well with luciferase reporter gene assay results. ? LXRs gene silencing significantly decreased hSULT2A1 expression. ? ChIP analysis suggested that genistein enhances hLXR? binding to the hSULT2A1 promoter.

  11. Mechanistic investigation of skin barrier perturbation induced by surfactants in the presence of humectants

    E-Print Network [OSTI]

    Ghosh, Saswata

    2007-01-01

    The stratum corneum (SC) of the skin functions as a barrier between the body and the environment. Surfactants such as Sodium Dodecyl Sulfate (SDS) are used in skin cleansers and in skin-care formulations because of their ...

  12. STORM: A General Model to Determine the Number and Adaptive Changes of Epithelial Stem Cells in Teleost, Murine and Human Intestinal Tracts

    E-Print Network [OSTI]

    Wang, Zhengyuan

    Intestinal stem cells play a pivotal role in the epithelial tissue renewal, homeostasis and cancer development. The lack of a general marker for intestinal stem cells across species has hampered analysis of stem cell number ...

  13. Triptolide, a diterpenoid triepoxide, induces antitumor proliferation via activation of c-Jun NH{sub 2}-terminal kinase 1 by decreasing phosphatidylinositol 3-kinase activity in human tumor cells

    SciTech Connect (OSTI)

    Miyata, Yoshiki [Department of Biochemistry and Molecular Biology, Tokyo University of Pharmacy and Life Science, School of Pharmacy, Hachioji, Tokyo 192-0392 (Japan); Sato, Takashi [Department of Biochemistry and Molecular Biology, Tokyo University of Pharmacy and Life Science, School of Pharmacy, Hachioji, Tokyo 192-0392 (Japan)]. E-mail: satotak@ps.toyaku.ac.jp; Ito, Akira [Department of Biochemistry and Molecular Biology, Tokyo University of Pharmacy and Life Science, School of Pharmacy, Hachioji, Tokyo 192-0392 (Japan)

    2005-11-04

    Triptolide, a diterpenoid triepoxide extracted from the Chinese herb Tripterygium wilfordii Hook f., exerts antitumorigenic actions against several tumor cells, but the intracellular target signal molecule(s) for this antitumorigenesis activity of triptolide remains to be identified. In the present study, we demonstrated that triptolide, in a dose-dependent manner, inhibited the proliferation of human fibrosarcoma HT-1080, human squamous carcinoma SAS, and human uterine cervical carcinoma SKG-II cells. In addition, triptolide was found to decrease phosphatidylinositol 3-kinase (PI3K) activity. A PI3K inhibitor, LY-294002, mimicked the triptolide-induced antiproliferative activity in HT-1080, SAS, and SKG-II cells. There was no change in the activity of Akt or protein kinase C (PKC), both of which are downstream effectors in the PI3K pathway. Furthermore, the phosphorylation of Ras, Raf, and mitogen-activated protein/extracellular signal-regulated kinase 1/2 was not modified in HT-1080 cells treated with triptolide. However, the phosphorylation of c-Jun NH{sub 2}-terminal kinase 1 (JNK1) was found to increase in both triptolide- and LY-294002-treated cells. Furthermore, the triptolide-induced inhibition of HT-1080 cell proliferation was not observed by JNK1 siRNA-treatment. These results provide novel evidence that PI3K is a crucial target molecule in the antitumorigenic action of triptolide. They further suggest a possible triptolide-induced inhibitory signal for tumor cell proliferation that is initiated by the decrease in PI3K activity, which in turn leads to the augmentation of JNK1 phosphorylation via the Akt and/or PKC-independent pathway(s). Moreover, it is likely that the activation of JNK1 is required for the triptolide-induced inhibition of tumor proliferation.

  14. THE DYNAMIC RELATIONSHIP BETWEEN LANGERHANS CELLS AND INTRAEPIDERMAL NERVE FIBERS IN THE MOUSE AND RAT FOOTPAD

    E-Print Network [OSTI]

    Doss, Argenia Lanisha Necole

    2011-12-31

    Skin disorders are often associated with immune and nervous system dysfunction. Intraepidermal nerve fibers (IENFs) detect mechanical, thermal, and noxious stimuli. Although immune cells such as mast and T cells can alter ...

  15. Analysis of Symmetry in the Anterior Human Dentition and its Application in the Evaluation and Correction of Postural Distortion in the Photographic Recording of Human Bite Marks 

    E-Print Network [OSTI]

    Aws, Ghassan

    Postural distortion of human bite marks on skin occurs when photographing a bite mark in a body position other than the position of the body at the time of biting. Postural distortion in the bite mark may introduce ...

  16. Skin-sparing Helical Tomotherapy vs 3D-conformal Radiotherapy for Adjuvant Breast Radiotherapy: In Vivo Skin Dosimetry Study

    SciTech Connect (OSTI)

    Capelle, Lisa; Warkentin, Heather; MacKenzie, Marc; Joseph, Kurian; Gabos, Zsolt; Pervez, Nadeem; Tankel, Keith; Chafe, Susan; Amanie, John; Ghosh, Sunita; Parliament, Matthew; Abdulkarim, Bassam

    2012-08-01

    Purpose: We investigated whether treatment-planning system (TPS)-calculated dose accurately reflects skin dose received for patients receiving adjuvant breast radiotherapy (RT) with standard three-dimensional conformal RT (3D-CRT) or skin-sparing helical tomotherapy (HT). Methods and Materials: Fifty patients enrolled in a randomized controlled trial investigating acute skin toxicity from adjuvant breast RT with 3D-CRT compared to skin-sparing HT, where a 5-mm strip of ipsilateral breast skin was spared. Thermoluminescent dosimetry or optically stimulated luminescence measurements were made in multiple locations and were compared to TPS-calculated doses. Skin dosimetric parameters and acute skin toxicity were recorded in these patients. Results: With HT there was a significant correlation between calculated and measured dose in the medial and lateral ipsilateral breast (r = 0.67, P<.001; r = 0.44, P=.03, respectively) and the medial and central contralateral breast (r = 0.73, P<.001; r = 0.88, P<.001, respectively). With 3D-CRT there was a significant correlation in the medial and lateral ipsilateral breast (r = 0.45, P=.03; r = 0.68, P<.001, respectively); the medial and central contralateral breast (r = 0.62, P=.001; r = 0.86, P<.001, respectively); and the mid neck (r = 0.42, P=.04, respectively). On average, HT-calculated dose overestimated the measured dose by 14%; 3D-CRT underestimated the dose by 0.4%. There was a borderline association between highest measured skin dose and moist desquamation (P=.05). Skin-sparing HT had greater skin homogeneity (homogeneity index of 1.39 vs 1.65, respectively; P=.005) than 3D-CRT plans. HT plans had a lower skin{sub V50} (1.4% vs 5.9%, respectively; P=.001) but higher skin{sub V40} and skin{sub V30} (71.7% vs 64.0%, P=.02; and 99.0% vs 93.8%, P=.001, respectively) than 3D-CRT plans. Conclusion: The 3D-CRT TPS more accurately reflected skin dose than the HT TPS, which tended to overestimate dose received by 14% in patients receiving adjuvant breast RT.

  17. The critical roles of pre-replicative complex (pre-RC) component, Cdc6, in DNA replication and checkpoint response in human cells

    E-Print Network [OSTI]

    Lau, Eric Kirk

    2008-01-01

    recognition complex (ORC)………………………………………5 B. Cell division231 Breast cancer cell type ORC Origin Recognition Complexby the timely arrival of the ORC complex proteins that bind

  18. Is the duration of skin disease visits decreasing in the united states?

    E-Print Network [OSTI]

    Davis, Scott A; Feldman, Steven R; Fleischer Jr., Alan B

    2015-01-01

    2). Visits for impetigo, psoriasis, and unspecified disorderneoplasm of skin (216.9) Psoriasis (696.1) unspecifiedneoplasm of skin (216.9) Psoriasis (696.1) Unspecified

  19. Effects of Low-Dose Alpha-Particle Irradiation in Human Cells: The Role of Induced Genes and the Bystander Effect. Final Technical Report (9/15/1998-5/31/2005)

    SciTech Connect (OSTI)

    Little, John B.

    2013-09-17

    This grant was designed to examine the cellular and molecular mechanisms for the bystander effect of radiation (initially described in this laboratory) whereby damage signals are passed from irradiated to non-irradiated cells in a population. These signals induce genetic effects including DNA damage, mutations and chromosomal aberrations in the nonirradiated cells. Experiments were carried out in cultured mammalian cells, primarily human diploid cells, irradiated with alpha particles. This research resulted in 17 publications in the refereed literature and is described in the Progress Report where it is keyed to the publication list. This project was initiated at the Harvard School of Public Health (HSPH) and continued in collaboration with students/fellows at Colorado State University (CSU) and the New Jersey Medical School (NJMS).

  20. Insulin-Like Growth Factor-Type 1 Receptor Inhibitor NVP-AEW541 Enhances Radiosensitivity of PTEN Wild-Type but Not PTEN-Deficient Human Prostate Cancer Cells

    SciTech Connect (OSTI)

    Isebaert, Sofie F.; Swinnen, Johannes V.; McBride, William H.; Haustermans, Karin M.

    2011-09-01

    Purpose: During the past decade, many clinical trials with both monoclonal antibodies and small molecules that target the insulin-like growth factor-type 1 receptor (IGF-1R) have been launched. Despite the important role of IGF-1R signaling in radioresistance, studies of such agents in combination with radiotherapy are lagging behind. Therefore, the aim of this study was to investigate the effect of the small molecule IGF-1R kinase inhibitor NVP-AEW541 on the intrinsic radioresistance of prostate cancer cells. Methods and Materials: The effect of NVP-AEW541 on cell proliferation, cell viability, IGF-1R signaling, radiosensitivity, cell cycle distribution, and double strand break repair was determined in three human prostate cancer cell lines (PC3, DU145, 22Rv1). Moreover, the importance of the PTEN pathway status was explored by means of transfection experiments with constitutively active Akt or inactive kinase-dead Akt. Results: NVP-AEW541 inhibited cell proliferation and decreased cell viability in a time-and dose-dependent manner in all three cell lines. Radiosensitization was observed in the PTEN wild-type cell lines DU145 and 22Rv1 but not in the PTEN-deficient PC3 cell line. NVP-AEW541-induced radiosensitization coincided with downregulation of phospho-Akt levels and high levels of residual double strand breaks. The importance of PTEN status in the radiosensitization effect was confirmed by transfection experiments with constitutively active Akt or inactive kinase-dead Akt. Conclusions: NVP-AEW541 enhances the effect of ionizing radiation in PTEN wild-type, but not in PTEN-deficient, prostate cancer cells. Proper patient selection based on the PTEN status of the tumor will be critical to the achievement of optimal results in clinical trials in which the combination of radiotherapy and this IGF-1R inhibitor is being explored.

  1. Modeling Stem Cell Induction Processes

    E-Print Network [OSTI]

    Gracio, Filipe

    Technology for converting human cells to pluripotent stem cell using induction processes has the potential to revolutionize regenerative medicine. However, the production of these so called iPS cells is still quite inefficient ...

  2. The Kauai Skin Cancer Study--1983 to 1992

    SciTech Connect (OSTI)

    Reizner, G.T. )

    1993-05-01

    The Kauai Skin Cancer Study began as a modest effort in 1983 to look at this island's skin cancer incidence. David Elpern MD, Kauai's only dermatologist at the time, was interested in the large number of these tumors in his practice. He first enlisted his office staff to help keep track of the numbers and type of these skin cancers. Along with this information, the basic demographic data on each patient was collected. These records became the first entries into what has become a decade-long project.

  3. Carmichael's Concise Review Microscopy is Only Skin Deep

    E-Print Network [OSTI]

    Heller, Eric

    Carmichael's Concise Review Microscopy is Only Skin Deep Stephen W. Carmichael Mayo Clinic. Coming Events 2011 EMAS 2011 May 15­19, 2011 Angers, France www.emas-web.net IUMAS-V May 22­27, 2011

  4. Mpemba paradox: Hydrogen bond memory and water-skin supersolidity

    E-Print Network [OSTI]

    Chang Q Sun

    2015-01-05

    Numerical reproduction of measurements, experimental evidence for skin super-solidity and hydrogen-bond memory clarified that Mpemba paradox integrates the heat emission-conduction-dissipation dynamics in the source-path-drain cycle system.

  5. Involvement of TGF-beta in skin photoaging

    E-Print Network [OSTI]

    Choi, Won Seon, 1975-

    2005-01-01

    The goal of this thesis study was to understand the role of TGF-[beta] in skin photoaging, especially in solar elastosis. Solar elastosis, the accumulation of elastotic material in the dermal extracelluar matrix, is a major ...

  6. ORIGINAL ARTICLE Bronchiectasis in Persons With Skin Lesions

    E-Print Network [OSTI]

    California at Berkeley, University of

    years of chronic cough underwent high-resolution computed tomography (CT); these scans were read reported chronic cough, CT evidence of bronchiectasis was found in 18 (67%) participants with skin lesions

  7. Physical Properties of Blue Shark Useful in Designing a Skinning Machine

    E-Print Network [OSTI]

    Physical Properties of Blue Shark Useful in Designing a Skinning Machine D. E. BROWN, R. PAUL SINGH for a machine to skin blue shark 2 · A wider market for blue shark products is being sought. For example, if removed in one piece the skin is of value for making leather. The machine proposed to skin blue shark

  8. Persistence of gamma-H2AX and 53BP1 foci in proliferating and nonproliferating human mammary epithelial cells after exposure to gamma-rays or iron ions

    SciTech Connect (OSTI)

    Groesser, Torsten; Chang, Hang; Fontenay, Gerald; Chen, James; Costes, Sylvain V.; Barcellos-Hoff, Mary Helen; Parvin, Bahram; Rydberg, Bjorn

    2010-12-22

    To investigate {gamma}-H2AX (phosphorylated histone H2AX) and 53BP1 (tumour protein 53 binding protein No. 1) foci formation and removal in proliferating and non-proliferating human mammary epithelial cells (HMEC) after exposure to sparsely and densely ionizing radiation under different cell culture conditions. HMEC cells were grown either as monolayers (2D) or in extracellular matrix to allow the formation of acinar structures in vitro (3D). Foci numbers were quantified by image analysis at various time points after exposure. Our results reveal that in non-proliferating cells under 2D and 3D cell culture conditions, iron-ion induced {gamma}-H2AX foci were still present at 72 h after exposure, although 53BP1 foci returned to control levels at 48 h. In contrast in proliferating HMEC, both {gamma}-H2AX and 53BP1 foci decreased to control levels during the 24-48 h time interval after irradiation under 2D conditions. Foci numbers decreased faster after {gamma}-ray irradiation and returned to control levels by 12 h regardless of marker, cell proliferation status, and cell culture condition. Conclusions: The disappearance of radiation induced {gamma}-H2AX and 53BP1 foci in HMEC have different dynamics that depend on radiation quality and proliferation status. Notably, the general patterns do not depend on the cell culture condition (2D versus 3D). We speculate that the persistent {gamma}-H2AX foci in iron-ion irradiated non-proliferating cells could be due to limited availability of double strand break (DSB) repair pathways in G0/G1-phase, or that repair of complex DSB requires replication or chromatin remodeling.

  9. Copyright c 2004 Tech Science Press MCB, vol.1, no.3, pp.169-180, 2004 Nonlinear Elastic and Viscoelastic Deformation of the Human Red Blood Cell with

    E-Print Network [OSTI]

    Dao, Ming

    of single cell mechanics under a variety of well-controlled stress-states. In this paper, we first crit contributions to deformation arising from the bending stiffness of the cell membrane and cytoskeleton. Finite not consid- ered. The in-plane shear modulus of the cell membrane was estimated from this approach to be 2

  10. RhoE interferes with Rb inactivation and regulates the proliferation and survival of the U87 human glioblastoma cell line

    SciTech Connect (OSTI)

    Poch, Enric [Departamento de Quimica, Bioquimica y Biologia Molecular, Universidad Cardenal Herrera-CEU, Valencia (Spain); Minambres, Rebeca [Laboratorio de Patologia Celular, Centro de Investigacion Principe Felipe, Valencia (Spain); Mocholi, Enric [Departamento de Quimica, Bioquimica y Biologia Molecular, Universidad Cardenal Herrera-CEU, Valencia (Spain); Ivorra, Carmen [Departamento de Quimica, Bioquimica y Biologia Molecular, Universidad Cardenal Herrera-CEU, Valencia (Spain); Perez-Arago, Amparo [Laboratorio de Patologia Celular, Centro de Investigacion Principe Felipe, Valencia (Spain); Guerri, Consuelo [Laboratorio de Patologia Celular, Centro de Investigacion Principe Felipe, Valencia (Spain); Perez-Roger, Ignacio [Departamento de Quimica, Bioquimica y Biologia Molecular, Universidad Cardenal Herrera-CEU, Valencia (Spain)]. E-mail: iperez@uch.ceu.es; Guasch, Rosa M. [Laboratorio de Patologia Celular, Centro de Investigacion Principe Felipe, Valencia (Spain)]. E-mail: guasch@cipf.es

    2007-02-15

    Rho GTPases are important regulators of actin cytoskeleton, but they are also involved in cell proliferation, transformation and oncogenesis. One of this proteins, RhoE, inhibits cell proliferation, however the mechanism that regulates this effect remains poorly understood. Therefore, we undertook the present study to determine the role of RhoE in the regulation of cell proliferation. For this purpose we generated an adenovirus system to overexpress RhoE in U87 glioblastoma cells. Our results show that RhoE disrupts actin cytoskeleton organization and inhibits U87 glioblastoma cell proliferation. Importantly, RhoE expressing cells show a reduction in Rb phosphorylation and in cyclin D1 expression. Furthermore, RhoE inhibits ERK activation following serum stimulation of quiescent cells. Based in these findings, we propose that RhoE inhibits ERK activation, thereby decreasing cyclin D1 expression and leading to a reduction in Rb inactivation, and that this mechanism is involved in the RhoE-induced cell growth inhibition. Moreover, we also demonstrate that RhoE induces apoptosis in U87 cells and also in colon carcinoma and melanoma cells. These results indicate that RhoE plays an important role in the regulation of cell proliferation and survival, and suggest that this protein may be considered as an oncosupressor since it is capable to induce apoptosis in several tumor cell lines.

  11. A component of green tea (-)-epigallocatechin-3-gallate, promotes apoptosis in T24 human bladder cancer cells via modulation of the PI3K/Akt pathway and Bcl-2 family proteins

    SciTech Connect (OSTI)

    Qin Jie [Department of Urology, First Affiliated Hospital, Medical College, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China); Xie Liping [Department of Urology, First Affiliated Hospital, Medical College, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China)]. E-mail: xielp@zjuem.zju.edu.cn; Zheng Xiangyi [Department of Urology, First Affiliated Hospital, Medical College, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China); Wang Yunbin [Department of Urology, First Affiliated Hospital, Medical College, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China); Bai Yu [Department of Urology, First Affiliated Hospital, Medical College, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China); Shen Huafeng [Department of Urology, First Affiliated Hospital, Medical College, Zhejiang University, Qingchun Road 79, Hangzhou 310003, Zhejiang Province (China); Li Longcheng [Department of Urology, University of California San Francisco and Veteran Affairs Medical Center San Francisco, San Francisco, CA (United States); Dahiya, Rajvir [Department of Urology, University of California San Francisco and Veteran Affairs Medical Center San Francisco, San Francisco, CA (United States)

    2007-03-23

    Bladder cancer is the fourth most common cancer in men and ninth most common in women. It has a protracted course of progression and is thus an ideal candidate for chemoprevention strategies and trials. This study was conducted to evaluate the chemopreventive/antiproliferative potential of (-)-epigallocatechin gallate (EGCG, the major phytochemical in green tea) against bladder cancer and its mechanism of action. Using the T24 human bladder cancer cell line, we found that EGCG treatment caused dose- and time-dependent inhibition of cellular proliferation and cell viability, and induced apoptosis. Mechanistically, EGCG inhibits phosphatidylinositol 3'-kinase/Akt activation that, in turn, results in modulation of Bcl-2 family proteins, leading to enhanced apoptosis of T24 cells. These findings suggest that EGCG may be an important chemoprevention agent for the management of bladder cancer.

  12. Method of forming a continuous polymeric skin on a cellular foam material

    DOE Patents [OSTI]

    Duchane, David V. (Los Alamos, NM); Barthell, Barry L. (Los Alamos, NM)

    1985-01-01

    Hydrophobic cellular material is coated with a thin hydrophilic polymer skin which stretches tightly over the outer surface of the foam but which does not fill the cells of the foam, thus resulting in a polymer-coated foam structure having a smoothness which was not possible in the prior art. In particular, when the hydrophobic cellular material is a specially chosen hydrophobic polymer foam and is formed into arbitrarily chosen shapes prior to the coating with hydrophilic polymer, inertial confinement fusion (ICF) targets of arbitrary shapes can be produced by subsequently coating the shapes with metal or with any other suitable material. New articles of manufacture are produced, including improved ICF targets, improved integrated circuits, and improved solar reflectors and solar collectors. In the coating method, the cell size of the hydrophobic cellular material, the viscosity of the polymer solution used to coat, and the surface tensin of the polymer solution used to coat are all very important to the coating.

  13. Co-localization and regulation of basic fibroblast growth factor and arginine vasopressin in neuroendocrine cells of the rat and human brain

    E-Print Network [OSTI]

    2010-01-01

    EGS: Designed human hypothalamus/ neurohypophysis studies,1b receptor mRNA in the hypothalamus and choroid plexus.ribonucleic acid in rat hypothalamus. Horm Metab Res 1995,

  14. Co-localization and regulation of basic fibroblast growth factor and arginine vasopressin in neuroendocrine cells of the rat and human brain

    E-Print Network [OSTI]

    2010-01-01

    from NIH NS/AG-91-03 (E.G.S. ), NINDS NS 27601 and NIA AGand manuscript drafting. EGS: Designed human hypothalamus/

  15. Identification of Genomic Predictors of Response to the CDK4/6 Inhibitor Palbociclib using the UCLATORL Panel of Human Cancer Cell Lines

    E-Print Network [OSTI]

    Conklin, Dylan Francis

    2013-01-01

    invasive epithelial ovarian cancer. Cancer treatment reviewsto breast and ovarian cancer. European journal of cancer 42,locus in advanced human ovarian-cancer. Int J Oncol 6, 129-

  16. tem cells are undifferentiated or "blank slate" cells from which other types of cells

    E-Print Network [OSTI]

    S tem cells are undifferentiated or "blank slate" cells from which other types of cells can arise. The defining characteristic of human stem cells is their ability to self-renew (provide an exact copy of themselves) while maintaining the potential to develop into other types of cells, such as blood, brain

  17. SU-E-T-68: Clinical Implementation of Total Skin Electron Beam Therapy: A New- York Presbyterian Hospital Experience

    SciTech Connect (OSTI)

    Afghan, M; Shih, R; Chen, H

    2014-06-01

    Purpose: Total skin electron beam therapy (TSET) is used in the treatment of rare skin diseases such as mycosis fungoides, the most common type of cutaneous T-cell lymphoma. We report our experience with clinical implementation of TSET. Methods: A modified six-dual-field irradiation technique was chosen to deliver TSET. A Varian Trilogy linear accelerator with a nominal 6 MeV beam using high dose rate total skin electron mode (HDTSe) was employed. The recommendations of AAPM task group report 23 were followed for the commissioning. An acrylic plate (energy degrader) of 3.2 mm depth was mounted on the HDTSe applicator. The nominal source to skin distance was set at 450 cm. The optimum tilt angle of the gantry was determined using NACP-02 ionization chamber embedded in certified therapy grade solid water. Percent depth dose measurements were performed using ionization chamber and radiochromic films embedded in solid water and anthropomorphic phantom. For absolute dose measurements, TG-51 formalism was employed. The dose distribution on the entire skin was measured by irradiating the anthropomorphic phantom, with TLDs attached, mimicking the real treatment. Results: The 3.2 mm acrylic plate mounted on the HDTSe applicator degraded the energy of the electron beam to 4.1 MeV in the treatment plane, located at an SSD of 450 cm. The optimum tilt angle was found to be ±20°. A single-dual field had a longitudinal uniformity, measured at a depth of dose maximum, of ±7% over a length of about 200 cm. For the entire treatment the multiplication factor was found to be 2.86. On the surface of the phantom, the dose varied from 108% to 93% of the prescription dose. Conclusion: We have successfully commissioned TSET meeting the guidelines of the TG report 23, and treated our first patient on February 25, 2014.

  18. (4-Methoxyphenyl)(3,4,5-trimethoxyphenyl)methanone inhibits tubulin polymerization, induces G{sub 2}/M arrest, and triggers apoptosis in human leukemia HL-60 cells

    SciTech Connect (OSTI)

    Magalhães, Hemerson I.F.; Wilke, Diego V.; Bezerra, Daniel P.; Cavalcanti, Bruno C.; Rotta, Rodrigo; Lima, Dênis P. de; Beatriz, Adilson; Moraes, Manoel O.; Diniz-Filho, Jairo; Pessoa, Claudia

    2013-10-01

    (4-Methoxyphenyl)(3,4,5-trimethoxyphenyl)methanone (PHT) is a known cytotoxic compound belonging to the phenstatin family. However, the exact mechanism of action of PHT-induced cell death remains to be determined. The aim of this study was to investigate the mechanisms underlying PHT-induced cytotoxicity. We found that PHT displayed potent cytotoxicity in different tumor cell lines, showing IC{sub 50} values in the nanomolar range. Cell cycle arrest in G{sub 2}/M phase along with the augmented metaphase cells was found. Cells treated with PHT also showed typical hallmarks of apoptosis such as cell shrinkage, chromatin condensation, phosphatidylserine exposure, increase of the caspase 3/7 and 8 activation, loss of mitochondrial membrane potential, and internucleosomal DNA fragmentation without affecting membrane integrity. Studies conducted with isolated tubulin and docking models confirmed that PHT binds to the colchicine site and interferes in the polymerization of microtubules. These results demonstrated that PHT inhibits tubulin polymerization, arrests cancer cells in G{sub 2}/M phase of the cell cycle, and induces their apoptosis, exhibiting promising anticancer therapeutic potential. - Highlights: • PHT inhibits tubulin polymerization. • PHT arrests cancer cells in G{sub 2}/M phase of the cell cycle. • PHT induces caspase-dependent apoptosis.

  19. The PIKfyve–ArPIKfyve–Sac3 triad in human breast cancer: Functional link between elevated Sac3 phosphatase and enhanced proliferation of triple negative cell lines

    SciTech Connect (OSTI)

    Ikonomov, Ognian C. Filios, Catherine Sbrissa, Diego Chen, Xuequn Shisheva, Assia

    2013-10-18

    Highlights: •We assess PAS complex proteins and phosphoinositide levels in breast cancer cells. •Sac3 and ArPIKfyve are markedly elevated in triple-negative breast cancer cells. •Sac3 silencing inhibits proliferation in triple-negative breast cancer cell lines. •Phosphoinositide profiles are altered in breast cancer cells. •This is the first evidence linking high Sac3 with breast cancer cell proliferation. -- Abstract: The phosphoinositide 5-kinase PIKfyve and 5-phosphatase Sac3 are scaffolded by ArPIKfyve in the PIKfyve–ArPIKfyve–Sac3 (PAS) regulatory complex to trigger a unique loop of PtdIns3P–PtdIns(3,5)P{sub 2} synthesis and turnover. Whereas the metabolizing enzymes of the other 3-phosphoinositides have already been implicated in breast cancer, the role of the PAS proteins and the PtdIns3P–PtdIns(3,5)P{sub 2} conversion is unknown. To begin elucidating their roles, in this study we monitored the endogenous levels of the PAS complex proteins in cell lines derived from hormone-receptor positive (MCF7 and T47D) or triple-negative breast cancers (TNBC) (BT20, BT549 and MDA-MB-231) as well as in MCF10A cells derived from non-tumorigenic mastectomy. We report profound upregulation of Sac3 and ArPIKfyve in the triple negative vs. hormone-sensitive breast cancer or non-tumorigenic cells, with BT cell lines showing the highest levels. siRNA-mediated knockdown of Sac3, but not that of PIKfyve, significantly inhibited proliferation of BT20 and BT549 cells. In these cells, knockdown of ArPIKfyve had only a minor effect, consistent with a primary role for Sac3 in TNBC cell proliferation. Intriguingly, steady-state levels of PtdIns(3,5)P{sub 2} in BT20 and T47D cells were similar despite the 6-fold difference in Sac3 levels between these cell lines. However, steady-state levels of PtdIns3P and PtdIns5P, both regulated by the PAS complex, were significantly reduced in BT20 vs. T47D or MCF10A cell lines, consistent with elevated Sac3 affecting directly or indirectly the homeostasis of these lipids in TNBC. Together, our results uncover an unexpected role for Sac3 phosphatase in TNBC cell proliferation. Database analyses, discussed herein, reinforce the involvement of Sac3 in breast cancer pathogenesis.

  20. Arsenite evokes IL-6 secretion, autocrine regulation of STAT3 signaling, and miR-21 expression, processes involved in the EMT and malignant transformation of human bronchial epithelial cells

    SciTech Connect (OSTI)

    Luo, Fei; Xu, Yuan; Ling, Min; Zhao, Yue; Xu, Wenchao; Liang, Xiao; Jiang, Rongrong; Wang, Bairu; Bian, Qian; Liu, Qizhan

    2013-11-15

    Arsenite is an established human carcinogen, and arsenite-induced inflammation contributes to malignant transformation of cells, but the molecular mechanisms by which cancers are produced remain to be established. The present results showed that, evoked by arsenite, secretion of interleukin-6 (IL-6), a pro-inflammatory cytokine, led to the activation of STAT3, a transcription activator, and to increased levels of a microRNA, miR-21. Blocking IL-6 with anti-IL-6 antibody and inhibiting STAT3 activation reduced miR-21 expression. For human bronchial epithelial cells, cultured in the presence of anti-IL-6 antibody for 3 days, the arsenite-induced EMT and malignant transformation were reversed. Thus, IL-6, acting on STAT3 signaling, which up-regulates miR-21in an autocrine manner, contributes to the EMT induced by arsenite. These data define a link from inflammation to EMT in the arsenite-induced malignant transformation of HBE cells. This link, mediated through miRNAs, establishes a mechanism for arsenite-induced lung carcinogenesis. - Highlights: • Arsenite evokes IL-6 secretion. • IL-6 autocrine mediates STAT3 signaling and up-regulates miR-21expression. • Inflammation is involved in arsenite-induced EMT.

  1. EGF-receptor phosphorylation and downstream signaling are activated by benzo[a]pyrene 3,6-quinone and benzo[a]pyrene 1,6-quinone in human mammary epithelial cells

    SciTech Connect (OSTI)

    Rodriguez-Fragoso, Lourdes [Facultad de Farmacia, Universidad Autonoma del Estado de Morelos, Avenida Universidad 1001 Col. Chamilpa, Cuernavaca 62210, Morelos (Mexico); Melendez, Karla; Hudson, Laurie G.; Lauer, Fredine T. [University of New Mexico, College of Pharmacy Toxicology Program, Albuquerque, New Mexico (United States); Burchiel, Scott W. [University of New Mexico, College of Pharmacy Toxicology Program, Albuquerque, New Mexico (United States)], E-mail: sburchiel@salud.unm.edu

    2009-03-15

    Benzo[a]pyrene (BaP) is activated by xenobiotic-metabolizing enzymes to highly mutagenic and carcinogenic metabolites. Previous studies in this laboratory have shown that benzo[a]pyrene quinones (BPQs), 1,6-BPQ and 3,6-BPQ, are able to induce epidermal growth factor receptor (EGFR) cell signaling through the production of reactive oxygen species. Recently, we have reported that BPQs have the potential to induce the expression of genes involved in numerous pathways associated with cell proliferation and survival in human mammary epithelial cells. In the present study we demonstrated that BPQs not only induced EGFR tyrosine autophosphorylation, but also induced EGFR-dependent tyrosine phosphorylation of phospholipase C-{gamma}1 and several signal transducers and activators of transcription (STATs). The effects of BPQs were evaluated in a model of EGF withdrawal in MCF10-A cells. We found that BPQs (1 {mu}M), induced EGFR tyrosine phosphorylation at positions Y845, Y992, Y1068, and Y1086. PLC-{gamma}1 phosphorylation correlated with the phosphorylation of tyrosine-Y992, a proposed docking site for PLC-{gamma}1 on the EGFR. Additionally, we found that BPQs induced the activation of STAT-1, STAT-3, STAT-5a and STAT-5b. STAT5 was shown to translocate to the nucleus following 3,6-BPQ and 1,6-BPQ exposures. Although the patterns of phosphorylation at EGFR, PLC-{gamma}1 and STATs were quite similar to those induced by EGF, an important difference between BPQ-mediated signaling of the EGFR was observed. Signaling produced by EGF ligand produced a rapid disappearance of EGFR from the cell surface, whereas BPQ signaling maintained EGFR receptors on the cell membrane. Thus, the results of these studies show that 1,6-BPQ and 3,6-BPQ can produce early events as evidenced by EGFR expression, and a prolonged transactivation of EGFR leading to downstream cell signaling pathways.

  2. Nuclear pygmy modes and the dynamics of the nuclear skin

    E-Print Network [OSTI]

    Nadia Tsoneva; Horst Lenske

    2012-11-05

    The information on pygmy resonances reveals new aspects on the isospin dynamics of the nucleus with important astrophysical consequences. In this connection, the precise knowledge of nuclear response functions plays a key role in the determination of photonuclear reactions cross sections which are of importance for the synthesis of heavy neutron-rich elements. For that purpose, a theoretical method based on density functional theory and multi-phonon approach is applied for investigations of nuclear excitations with different multipolarities and energies in stable and exotic nuclei. The possible relation of low-energy modes to the properties of neutron or proton skins is systematically investigated for isotonic and isotopic chains. Our studies of dipole and quadrupole response functions and the corresponding transition densities indicate new pygmy dipole and pygmy quadrupole resonances, describing oscillations of the nuclear skin. Also, the presence of skins is found to affect the magnetic response of nuclei.

  3. Interference of silibinin with IGF-1R signalling pathways protects human epidermoid carcinoma A431 cells from UVB-induced apoptosis

    SciTech Connect (OSTI)

    Liu, Weiwei; Otkur, Wuxiyar; Li, Lingzhi; Wang, Qiong; He, Hao; Zang, Linghe; Hayashi, Toshihiko [China–Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016 (China)] [China–Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016 (China); Tashiro, Shin-ichi [Institute for Clinical and Biomedical Sciences, Kyoto 603-8072 (Japan)] [Institute for Clinical and Biomedical Sciences, Kyoto 603-8072 (Japan); Onodera, Satoshi [Department of Clinical and Biomedical Sciences, Showa Pharmaceutical University, Tokyo 194-8543 (Japan)] [Department of Clinical and Biomedical Sciences, Showa Pharmaceutical University, Tokyo 194-8543 (Japan); Xia, Mingyu [China–Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016 (China)] [China–Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016 (China); Ikejima, Takashi, E-mail: ikejimat@vip.sina.com [China–Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016 (China)] [China–Japan Research Institute of Medical and Pharmaceutical Sciences, Shenyang Pharmaceutical University, Shenyang 110016 (China)

    2013-03-08

    Highlights: ? Silibinin protects A431 cells from UVB irradiation-induced apoptosis. ? Up-regulation of the IGF-1R-JNK/ERK pathways by UVB induces cell apoptosis. ? Silibinin inhibits IGF-1R pathways to repress caspase-8-mediated apoptosis. -- Abstract: Ultraviolet B (UVB) from sunlight is a major cause of cutaneous lesion. Silibinin, a traditional hepatic protectant, elicits protective effects against UVB-induced cellular damage. In A431 cells, the insulin-like growth factor-1 receptor (IGF-1R) was markedly up-regulated by UVB irradiation. The activation of the IGF-1R signalling pathways contributed to apoptosis of the cells rather than rescuing the cells from death. Up-regulated IGF-1R stimulated downstream mitogen-activated protein kinases (MAPKs), such as c-Jun N-terminal kinases (JNK) and extracellular signal-regulated protein kinases 1/2 (ERK1/2). The subsequent activation of caspase-8 and caspase-3 led to apoptosis. The activation of IGF-1R signalling pathways is the cause of A431 cell death. The pharmacological inhibitors and the small interfering RNA (siRNA) targeting IGF-1R suppressed the downstream activation of JNK/ERK-caspases to help the survival of the UVB-irradiated A431 cells. Indeed, silibinin treatment suppressed the IGF-1R-JNK/ERK pathways and thus protected the cells from UVB-induced apoptosis.

  4. Transcriptional functions of the corepressor Sin3A in skin

    E-Print Network [OSTI]

    Cox, Claire

    2013-03-12

    by the fact that disruption of their expression leads to skin disease. For example, a number of members of the S100 gene family have been observed to be up-regulated in psoriasis and skin cancer [Hoffjan and Stemmler, 2007]. Disruption of the expression... of genes in the Flg-like, LCE and SPRR families has been linked with the development of atopic dermatitis and psoriasis as well as ichthyosis vulgaris [de Guzman Strong et al., 2010]. A number of transcription factors have been implicated in the control...

  5. Meeting Report. Assessing Human Germ-Cell Mutagenesis in the Post-Genome Era: A Celebration of the Legacy of William Lawson (Bill) Russell

    E-Print Network [OSTI]

    2006-01-01

    landmark Human Genome Project (HGP) and its relationship toMoyzis explained that the HGP initially arose out of concernHowever, with time, the HGP changed its focus to concentrate

  6. Inhibition of KCa3.1 by depolarisation and 2-aminoethoxydiphenyl borate (2-APB) during Ca2+ release activated Ca2+ (CRAC) entry in human erythroleukemia (HEL) cells: Implications for the interpretation of 2-APB inhibition of CRAC entry

    E-Print Network [OSTI]

    Littlechild, Robert; Zaidman, Nathalie; Khodaverdi, Darren; Mason, Michael James

    2014-12-23

    target effects began to mount up. These include block of endosomal Ca2+ pumps [9], voltage-gated K+ channels [10], the non-selective cation channel TRPM7 [11], a Mg2+-inhibited K+ conductance described in human erythroleukemia (HEL) cells [12... /2 is the concentration of 2-APB that yields half maximal inhibition, and n is the apparent cooperativity of the process. 2.7 Statistical analysis and presentation Data are presented as the mean ± the SEM. All error bars on graph symbols denote the SEM...

  7. Hypothesis: {open_quotes}Rogue cell{close_quotes}-type chromosomal damage in lymphocytes is associated with infection with the JC human polyoma virus and has implications for oncopenesis

    SciTech Connect (OSTI)

    Neel, J.V.; Glover, T.; Burgess, A. [Univ. of Michigan Medical School, Ann Arbor, MI (United States)] [and others] [Univ. of Michigan Medical School, Ann Arbor, MI (United States); and others

    1996-04-02

    The hemagglutination inhibition antibody titers against the JC and BK polyoma viruses (JCV and BKV, respectively) are significantly elevated in individuals exhibiting {open_quotes}rogue{close_quotes} cells among their cultured lymphocytes. However, the elevation is so much greater with respect to JCV that the BKV elevation could readily be explained by cross reactivity to the capsid protein of these two closely related viruses. The JCV exhibits highly sequence homology with the simian papovavirus, simian virus 40 (SV40), and inoculation of human fetal brain cells with JCV produces polyploidy and chromosomal damage very similar to that produced by SV40. We suggest, by analogy with the effects of SV40, that these changes are due to the action of the viral large tumor antigen, a pluripotent DNA binding protein that acts in both transcription and replication. The implications of these findings for oncogenesis are briefly discussed. 45 refs., 1 fig., 3 tabs.

  8. Assay for mutagenesis in heterozygous diploid human lymphoblasts

    DOE Patents [OSTI]

    Skopek, Thomas R. (Somerville, MA); Liber, Howard L. (Brookline, MA); Penman, Bruce W. (Cambridge, MA); Thilly, William G. (Cambridge, MA); Hoppe, IV, Henry (Arlington, MA)

    1981-01-01

    An assay is disclosed for determining mutagenic damage caused by the administration of a known or suspected mutagen to diploid human lymphoblastoid cell lines. The gene locus employed for this assay is the gene for thymidine kinase, uridine kinase, or cytidine deaminase. Since human lymphoblastoid cells contain two genes for these enzymes, heterozygotes of human lymphoblastoid cells are used in this assay.

  9. Human energy

    E-Print Network [OSTI]

    Sawyer, Suzana

    2010-01-01

    This is the power of human energy that Chevron neverExperience the power of ‘‘Human Energy’’: S. Sawyer (&)s voice returns: ‘‘This is the power of human energy. ’’ In

  10. Skin friction and pressure: the "footprints" of turbulence

    E-Print Network [OSTI]

    Protas, Bartosz

    been a flurry of activity in controlling both laminar and turbulent flows in certain idealized settings, and to begin to shed light on how to control fluid flow in practical engineering applications with modelSkin friction and pressure: the "footprints" of turbulence Thomas R. Bewley and Bartosz Protas Flow

  11. MOBILE PHONE USE AND TEMPORAL SKIN HEAT SENSATION

    E-Print Network [OSTI]

    Boyer, Edmond

    in the phone by the battery currents and running of the radiofrequency (RF) electronic circuits measured the temperature of the temporal skin due to GSM 1800 MHz MP radiated power (125 mW). We suppressed of the heat produced in the phone by the battery currents and running of the radiofrequency (RF) electronic

  12. ORIGINAL ARTICLE Co-habiting amphibian species harbor unique skin

    E-Print Network [OSTI]

    McKenzie, Valerie

    ; microbiome Introduction All species of plants and animals harbor assem- blages of microbes the microbiomes of different species of animals, and fewer still have examined animals in the wild. We sampled: microbe­microbe and microbe­host interactions Keywords: amphibian; skin; bacteria; host specific

  13. OUTLIER ESTIMATION AND DETECTION APPLICATION TO SKIN LESION CLASSIFICATION

    E-Print Network [OSTI]

    OUTLIER ESTIMATION AND DETECTION APPLICATION TO SKIN LESION CLASSIFICATION S. Sigurdsson£ , J the project Signal and Image Processing for Telemedicine (SITE). Outliers are defined as an input pattern be rewritten as Ô´ Рܵ Ô¼´ Рܵ´½ ¬ µ·¬ (2) where ¬ ¼ ½ ´ ½µ . 3. NETWORK ARCHITECTURE AND INFERENCE

  14. Criminal Skins: Tattoos and Modern Architecture in the

    E-Print Network [OSTI]

    Canales, Jimena

    Criminal Skins: Tattoos and Modern Architecture in the Work of Adolf Loos by JIMENA CANALES, `Ornament and Crime', decisively linked unornamented architecture with the culture of modernity and, in so doing, became one of the key formulations of modern architecture.1 To a great extent, the essay's force

  15. SKIN CANCER INSTITUTE THE CANCER INSTITUTES AT NORTHWESTERN MEDICINE

    E-Print Network [OSTI]

    Chisholm, Rex L.

    of specimens with unclear diagnoses sent from throughout the United States. Our pathologists are thus some than 75 percent of skin cancer deaths. It is now estimated that one in every 55 people will be diagnosed with melanoma. The incidence also is rising at a rate faster than that of the seven most common

  16. Neutron skin uncertainties of Skyrme energy density functionals

    E-Print Network [OSTI]

    M. Kortelainen; J. Erler; W. Nazarewicz; N. Birge; Y. Gao; E. Olsen

    2013-07-16

    Background: Neutron-skin thickness is an excellent indicator of isovector properties of atomic nuclei. As such, it correlates strongly with observables in finite nuclei that depend on neutron-to-proton imbalance and the nuclear symmetry energy that characterizes the equation of state of neutron-rich matter. A rich worldwide experimental program involving studies with rare isotopes, parity violating electron scattering, and astronomical observations is devoted to pinning down the isovector sector of nuclear models. Purpose: We assess the theoretical systematic and statistical uncertainties of neutron-skin thickness and relate them to the equation of state of nuclear matter, and in particular to nuclear symmetry energy parameters. Methods: We use the nuclear superfluid Density Functional Theory with several Skyrme energy density functionals and density dependent pairing. To evaluate statistical errors and their budget, we employ the statistical covariance technique. Results: We find that the errors on neutron skin increase with neutron excess. Statistical errors due to uncertain coupling constants of the density functional are found to be larger than systematic errors, the latter not exceeding 0.06 fm in most neutron-rich nuclei across the nuclear landscape. The single major source of uncertainty is the poorly determined slope L of the symmetry energy that parametrizes its density dependence. Conclusions: To provide essential constraints on the symmetry energy of the nuclear energy density functional, next-generation measurements of neutron skins are required to deliver precision better than 0.06 fm.

  17. Viral oncogenesis and cell differentiation

    SciTech Connect (OSTI)

    Diamond, L.; Wolman, S.R.

    1989-01-01

    This book covers the following topics: Retroviruses, Human lymphotropic viruses, Oncogenes, Hematopoiesis: Normal and Abnormal, Growth and differentiation of normal and malignant cells, Molecular and genetic control of cell proliferation.

  18. Chromosomal translocation in a human leukemic stem-cell line disrupts the T-cell antigen receptor. delta. -chain diversity region and results in a previously unreported fusion transcript

    SciTech Connect (OSTI)

    Begley, C.G.; Aplan, P.D.; Davey, M.P.; Nakahara, K.; Tchorz, K.; Cohen, D.I.; Waldmann, T.A.; Kirsch, I.R. (National Institutes of Health, Bethesda, MD (USA)); Kurtzberg, J.; Hershfield, M.S.; Haynes, B.F. (Duke Univ. Medical Center, Durham, NC (USA))

    1989-03-01

    The authors have studied a leukemic stem-cell line, DU.528, that is able to differentiate into myeloid and lymphoid cells. The leukemic cells have a translocation between chromosomes 1 and 14, t(1;14)(p33;q11), which they have molecularly cloned and sequenced. Initial screening used joining (J)-segment probes from the T-cell receptor (TCR) {alpha}- and {delta}-chain loci. In apparent concert with the translocation, a deletion has occurred between {delta}-chain diversity (D)-region genes D{sub {delta}1} and D{sub {delta}2}. The nature of the D{sub {delta}1}-D{sub {delta}2} deletional event implicates a lymphoid recombinase in the mechanism of the translocation. As a consequence of the translocation, an unusual fusion transcript was generated. Probes from chromosome 1 detected a previously unreported transcript in RNA from both the cell line and the patient. A chromosome 14 probe identified the same transcript, thus confirming a fusion transcript derived from both chromosomes 1 and 14. This translocation may identify a gene for which they propose the name SCL (stem-cell leukemia) that is important for hemopoietic development and oncogenesis and that has been disrupted or altered in this stem-cell line.

  19. Activation of the PI3K-Akt pathway by human T cell leukemia virus type 1 (HTLV-1) oncoprotein Tax increases Bcl3 expression, which is associated with enhanced growth of HTLV-1-infected T cells

    SciTech Connect (OSTI)

    Saito, Kousuke; Saito, Mineki; Taniura, Naoko; Okuwa, Takako; Ohara, Yoshiro

    2010-08-01

    Bcl3 is a member of the I{kappa}B family that regulates genes involved in cell proliferation and apoptosis. Recent reports indicated that Bcl3 is overexpressed in HTLV-1-infected T cells via Tax-mediated transactivation, and acts as a negative regulator of viral transcription. However, the role of Bcl3 in cellular signal transduction and the growth of HTLV-1-infected T cells have not been reported. In this study, we showed that the knockdown of Bcl3 by short hairpin RNA inhibited the growth of HTLV-1-infected T cells. Although phosphatidylinositol-3 kinase (PI3K) inhibitor reduced Bcl3 expression, inactivation of glycogen synthase kinase 3 (GSK3), an effector kinase of the PI3K/Akt signaling pathway, restored Bcl3 expression in Tax-negative but not in Tax-positive T cells. Our results indicate that the overexpression of Bcl3 in HTLV-1-infected T cells is regulated not only by transcriptional but also by post-transcriptional mechanisms, and is involved in overgrowth of HTLV-1-infected T cells.

  20. HandWave : design and manufacture of a wearable wireless skin conductance sensor and housing

    E-Print Network [OSTI]

    Strauss, Marc D

    2005-01-01

    This thesis report details the design and manufacture of HandWave, a wearable wireless Bluetooth skin conductance sensor, and dedicated housing. The HandWave collects Electrodermal Activity (EDA) data by measuring skin ...

  1. Design and optimization of actuation mechanisms for rapid skin closure device

    E-Print Network [OSTI]

    Erickson, Andrew T. (Andrew Thomas)

    2012-01-01

    Innovative mechanism designs were explored for the actuation of critical components in a novel rapid skin closure device used to close long surgical incisions. The rapid skin closure device is designed to speed up the wound ...

  2. Human radiation studies: Remembering the early years: Oral history of cell biologist Don Francis Petersen, Ph.D., conducted November 29, 1994

    SciTech Connect (OSTI)

    1995-08-01

    This report is a transcript of an interview of Dr. Don Francis Petersen by representatives of the US DOE Office of Human Radiation Experiments. Dr. Petersen was selected for this interview because of his long research career at Los Alamos and his knowledge of the Atomic Energy Commission`s biomedical program. Dr. Petersen did not personally conduct research on human subjects. After a brief biographical sketch Dr. Petersen discusses his remembrances of the early use of radionuclides as biological tracers, aspects of nuclear weapons testing in the 1940`s and 1950`s including fallout studies, the means by which research projects were approved, use of humans in the whole-body counter, and the Health Division Biomedical responsibilities.

  3. FRACTURE OF SKIN-STIFFENER INTERSECTIONS IN COMPOSITE WIND TURBINE BLADE STRUCTURES

    E-Print Network [OSTI]

    FRACTURE OF SKIN-STIFFENER INTERSECTIONS IN COMPOSITE WIND TURBINE BLADE STRUCTURES by Darrin John..................................................................4 Stiffener Design Considerations

  4. Growth factor regulation of proliferation and survival of multipotential stromal cells

    E-Print Network [OSTI]

    Rodrigues, Melanie

    Multipotential stromal cells (MSCs) have been touted to provide an alternative to conservative procedures of therapy, be it heart transplants, bone reconstruction, kidney grafts, or skin, neuronal and cartilage repair. A ...

  5. Skin Cancer: A Young Person's Disease By Lauren Duffy (B.S. Communication, Journalism '14)

    E-Print Network [OSTI]

    Massachusetts at Amherst, University of

    Skin Cancer: A Young Person's Disease By Lauren Duffy (B.S. Communication, Journalism '14 is that this behavior is extremely unhealthy and risky for their bodies, specifically their skin. Skin cancer is the most common form of cancer found in young adults and second most common cancer found in adolescents

  6. How to Care for Your Wound After It's Treated With DERMABOND* Topical Skin Adhesive

    E-Print Network [OSTI]

    Kay, Mark A.

    How to Care for Your Wound After It's Treated With DERMABOND* Topical Skin Adhesive DERMABOND* Topical Skin Adhesive (2-octyl cyanoacrylate) is a sterile, liquid skin adhesive that holds wound edges · If your wound is bandaged, keep the bandage dry. · Replace the dressing daily until the adhesive film has

  7. Activation of Human ?? T Cells by Cytosolic Interactions of BTN3A1 with Soluble Phosphoantigens and the Cytoskeletal Adaptor Periplakin

    E-Print Network [OSTI]

    Rhodes, David A.; Chen, Hung-Chang; Price, Amanda J.; Keeble, Anthony H.; Davey, Martin S.; James, Leo C.; Eberl, Matthias; Trowsdale, John

    2015-01-30

    . 2009. Cellular expression, trafficking, and function of two isoforms of human ULBP5/ RAET1G. PLoS ONE 4: e4503. 13. Biris, N., Y. Yang, A. B. Taylor, A. Tomashevski, M. Guo, P. J. Hart, F. Diaz- Griffero, and D. N. Ivanov. 2012. Structure of the rhesus...

  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. Probing of a human proteome microarray with a recombinant pathogen protein reveals a novel mechanism by which hookworms suppress B cell receptor signaling

    E-Print Network [OSTI]

    Tribolet, Leon; Cantacessi, Cinzia; Pickering, Darren; Navarro, Severine; Doolan, Denise; Trieu, Angela; Fei, Huang; Chao, Yang; Hofmann, Andreas; Gasser, Robin; Giacomin, Paul; Loukas, Alex

    2014-08-19

    , Sigma). 154 155 Flow cytometry and cell sorting 156 Cells were stained with fluorophore-conjugated antibodies against CD19 (HIB19), CD20 (2H7), 157 CD3 (OKT3), CD4 (OKT4), CD14 (61D3) CD69 (FN50), HLA-DR (LN3), CD80 (2D10.4), CD86 158 (IT2.2) (all... . Controlling the false discovery rate in 462 behavior genetics research. Behav Brain Res 2001; 125:279-84. 463 30. Boyle EI, Weng S, Gollub J, et al. GO::TermFinder--open source software for accessing Gene 464 Ontology information and finding significantly...

  10. European trends in the frequency of original research in acne vulgaris, rosacea, dermatitis, psoriasis, skin cancer, and skin infections, 1970-2010

    E-Print Network [OSTI]

    Choi, Young M; Garcha, Jaspreet K; Wu, Jashin J

    2015-01-01

    S, Wolk K. Three decades of psoriasis research: where has itrosacea, dermatitis, psoriasis, skin cancer, and skinnotably, we found that psoriasis publications peaked around

  11. Lithium Ion Battery Performance of Silicon Nanowires With Carbon Skin

    SciTech Connect (OSTI)

    Bogart, Timothy D.; Oka, Daichi; Lu, Xiaotang; Gu, Meng; Wang, Chong M.; Korgel, Brian A.

    2013-12-06

    Silicon (Si) nanomaterials have emerged as a leading candidate for next generation lithium-ion battery anodes. However, the low electrical conductivity of Si requires the use of conductive additives in the anode film. Here we report a solution-based synthesis of Si nanowires with a conductive carbon skin. Without any conductive additive, the Si nanowire electrodes exhibited capacities of over 2000 mA h g-1 for 100 cycles when cycled at C/10 and over 1200 mA h g-1 when cycled more rapidly at 1C against Li metal.. In situ transmission electron microscopy (TEM) observation reveals that the carbon skin performs dual roles: it speeds lithiation of the Si nanowires significantly, while also constraining the final volume expansion. The present work sheds light on ways to optimize lithium battery performance by smartly tailoring the nanostructure of composition of materials based on silicon and carbon.

  12. Probing the neutron skin thickness in collective modes of excitation

    E-Print Network [OSTI]

    Nils Paar; Andrea Horvat

    2014-01-13

    Nuclear collective motion provides valuable constraint on the size of neutron-skin thickness and the properties of nuclear matter symmetry energy. By employing relativistic nuclear energy density functional (RNEDF) and covariance analysis related to $\\chi^2$ fitting of the model parameters, relevant observables are identified for dipole excitations, which strongly correlate with the neutron-skin thickness $(r_{np})$, symmetry energy at saturation density $(J)$ and slope of the symmetry energy $(L)$. Using the RNEDF framework and experimental data on pygmy dipole strength ($^{68}$Ni, $^{132}$Sn, $^{208}$Pb) and dipole polarizability ($^{208}$Pb), it is shown how the values of $J$, and $L$, and $r_{np}$ are constrained. The isotopic dependence of moments associated to dipole excitations in $^{116-136}$Sn shows that the low-energy dipole strength and polarizability in neutron-rich nuclei display strong sensitivity to the symmetry energy parameter $J$, more pronounced than in isotopes with moderate neutron-to-proton number ratios.

  13. Emergence of pygmy dipole resonances: Magic numbers and neutron skins

    E-Print Network [OSTI]

    Tsunenori Inakura; Takashi Nakatsukasa; Kazuhiro Yabana

    2011-06-18

    The pygmy dipole resonances (PDR) for even-even nuclei in 8=emerges by showing a peak of the E1 strength at energies less than 10 MeV. The E1 strength of the PDR strongly depends on the position of the Fermi level and shows a clear correlation with the occupation of the orbits with the orbital angular momenta less than 3\\hbar (l =< 2). We also found a strong correlation between the isotopic dependence of the neutron skin thickness and the pygmy dipole strength. The fraction of the energy weighted strength exhausted by the PDR and the neutron skin thickness show a linear correlation with the universal rate of about 0.2/fm.

  14. Adipose-Derived Perivascular Stem Cells Heal Critical Size Mouse Calvarial Defects

    E-Print Network [OSTI]

    Megerdichian, Silva

    2013-01-01

    stromal  cells  derived  from  the  infrapatellar  fat  C.  M.   et  al.  Adipose-­?derived  adult  stromal  cells  Human  bone  marrow-­? derived  mesenchymal  stem  cells  

  15. Nitric oxide-releasing sulindac is a novel skin cancer chemopreventive agent for UVB-induced photocarcinogenesis

    SciTech Connect (OSTI)

    Chaudhary, Sandeep C.; Singh, Tripti; Kapur, Puneet; Weng, Zhiping; Arumugam, Aadithya; Elmets, Craig A. [Department of Dermatology, University of Alabama at Birmingham, 1530 3rd Avenue South, VH509, Birmingham, AL 35294-0019 (United States); Kopelovich, Levy [Division of Cancer Prevention, National Cancer Institute, 6130 Executive Blvd, Suite 2114, Bethesda, MD 20892 (United States); Athar, Mohammad, E-mail: mathar@uab.edu [Department of Dermatology, University of Alabama at Birmingham, 1530 3rd Avenue South, VH509, Birmingham, AL 35294-0019 (United States)

    2013-05-01

    Nitric oxide (NO)-releasing non-steroidal anti-inflammatory drugs (NO-NSAIDs) which have been synthesized to reduce gastro-intestinal and cardiovascular toxicities of NSAIDs, possess anti-proliferative, pro-apoptotic and anti-cancer activities. Here, we show that NO-sulindac inhibited UVB-induced skin tumorigenesis in SKH-1 hairless mice. Topical application of NO-sulindac reduced tumor incidence, number (p < 0.05) and volume (p < 0.005) as compared to UVB (alone)-irradiated vehicle-treated mice. An increase in TUNEL-positive cells in skin lesions was accompanied by the enhanced Bax:Bcl-2 ratio. The expression of pro-apoptotic Bax was increased whereas anti-apoptotic Bcl-2 reduced. However, proliferation was identified as the major target of NO-sulindac in this study. A reduced expression of PCNA and cyclin D1 associated with the dampening of cell cycle progression was observed. The mechanism of this inhibition was related to the reduction in UVB-induced Notch signaling pathway. UVB-induced inflammatory responses were diminished by NO-sulindac as observed by a remarkable reduction in the levels of phosphorylated MAP Kinases Erk1/2, p38 and JNK1/2. In this regard, NO-sulindac also inhibited NF?B by enhancing I?B? as evidenced by the reduced expression of iNOS and COX-2, the direct NF?B transcription target proteins. NO-sulindac significantly diminished the progression of benign lesions to invasive carcinomas by suppressing the tumor aggressiveness and retarding epithelial–mesenchymal transition. A marked decrease in the expression of mesenchymal markers such as Fibronectin, N-cadherin, SNAI, Slug and Twist and an increase in epithelial cell polarity marker E-cadherin were noted in NO-sulindac-treated tumors. Our data suggest that NO-sulindac is a potent inhibitor of UVB-induced skin carcinogenesis and acts by targeting proliferation-regulatory pathways. - Highlights: ? NO-sulindac is a potent chemopreventive agent for UVB-induced skin cancer. ? NO-sulindac effectively blocks proliferation. ? NO-sulindac targets Notch and RXR-PI3k/Akt pathway to achieve anti-tumor efficacy.

  16. The eukaryotic translation elongation factor eEF1A2 induces neoplastic properties and mediates tumorigenic effects of ZNF217 in precursor cells of human ovarian carcinomas

    SciTech Connect (OSTI)

    Sun, Yu; Wong, Nicholas; Guan, Yinghui; Salamanca, Clara M.; Cheng, Jung Chien; Lee, Jonathan M.; Gray, Joe W.; Auersperg, Nelly

    2008-04-25

    Ovarian epithelial carcinomas (OEC) frequently exhibit amplifications at the 20q13 locus which is the site of several oncogenes, including the eukaryotic elongation factor EEF1A2 and the transcription factor ZNF217. We reported previously that overexpressed ZNF217 induces neoplastic characteristics in precursor cells of OEC. Unexpectedly, ZNF217, which is a transcriptional repressor, enhanced expression of eEF1A2. In this study, array comparative genomic hybridization, single nucleotide polymorphism and Affymetrix analysis of ZNF217-overexpressing cell lines confirmed consistently increased expression of eEF1A2 but not of other oncogenes, and revealed early changes in EEF1A2 gene copy numbers and increased expression at crisis during immortalization. We defined the influence of eEF1A2 overexpression on immortalized ovarian surface epithelial cells, and investigated interrelationships between effects of ZNF217 and eEF1A2 on cellular phenotypes. Lentivirally induced eEF1A2 overexpression caused delayed crisis, apoptosis resistance and increases in serum-independence, saturation densities, and anchorage independence. siRNA to eEF1A2 reversed apoptosis resistance and reduced anchorage independence in eEF1A2-overexpressing lines. Remarkably, siRNA to eEF1A2 was equally efficient in inhibiting both anchorage independence and resistance to apoptosis conferred by ZNF217 overexpression. Our data define neoplastic properties that are caused by eEF1A2 in nontumorigenic ovarian cancer precursor cells, and suggest that eEF1A2 plays a role in mediating ZNF217-induced neoplastic progression.

  17. Human Fertilisation and Embryology Bill A briefing for MPs on the need for research involving human and human admixed embryos.

    E-Print Network [OSTI]

    Rambaut, Andrew

    cells for research. The Bill allows research, under license, on four types of HAEs, which contain bothHuman Fertilisation and Embryology Bill A briefing for MPs on the need for research involving human and human admixed embryos. Prepared by the Academy of Medical Sciences, the Medical Research Council

  18. Background Information 1. What are stem cells?

    E-Print Network [OSTI]

    Rambaut, Andrew

    Background Information 1. What are stem cells? 2. What might stem cell research achieve? 3. Why we need to continue research using embryonic stem cells? 4. Time taken for discoveries 5. Examples of stem cell therapies in clinical trials 6. Patentability of human embryonic stem cell therapies 7. Creation

  19. mRNA Display selection of a novel activated leukocyte cell adhesion molecule (ALCAM) binding protein from a modified combinatorial protein library based on the tenth domain of human fibronection III (10FnIII)

    E-Print Network [OSTI]

    Park, Ann

    2015-01-01

    Activated Leukocyte Cell Adhesion Molecule (ALCAM)-Positiveof activated leukocyte-cell adhesion molecule (ALCAM), a CD6Activated leukocyte cell adhesion molecule (CD166/ALCAM):

  20. Human MSH2 protein

    DOE Patents [OSTI]

    de la Chapelle, Albert (Helsingfors, FI); Vogelstein, Bert (Baltimore, MD); Kinzler, Kenneth W. (Baltimore, MD)

    1997-01-01

    The human MSH2 gene, responsible for hereditary non-polyposis colorectal cancer, was identified by virtue of its homology to the MutS class of genes, which are involved in DNA mismatch repair. The sequence of cDNA clones of the human gene are provided, and the sequence of the gene can be used to demonstrate the existence of germ line mutations in hereditary non-polyposis colorectal cancer (HNPCC) kindreds, as well as in replication error.sup.+ (RER.sup.+) tumor cells.

  1. Human MSH2 protein

    DOE Patents [OSTI]

    Chapelle, A. de la; Vogelstein, B.; Kinzler, K.W.

    1997-01-07

    The human MSH2 gene, responsible for hereditary non-polyposis colorectal cancer, was identified by virtue of its homology to the MutS class of genes, which are involved in DNA mismatch repair. The sequence of cDNA clones of the human gene are provided, and the sequence of the gene can be used to demonstrate the existence of germ line mutations in hereditary non-polyposis colorectal cancer (HNPCC) kindreds, as well as in replication error{sup +} (RER{sup +}) tumor cells. 19 figs.

  2. Steady-state directional diffuse reflectance and fluorescence of human skin

    E-Print Network [OSTI]

    Pilon, Laurent

    .3660, 170.7050, 290.7050, 300.2530. 1. Introduction Fluorescence is the physical phenomenon in which light

  3. Systematic Identification and Characterization of Novel Human Skin-Associated Genes Encoding Membrane and Secreted Proteins

    E-Print Network [OSTI]

    2013-01-01

    atopic dermatitis and psoriasis vulgaris. J Allergy Clinsystem that is active in psoriasis and promotes keratinocyteatopic dermatitis and psoriasis. Br J Dermatol 15. Kim SJ,

  4. Systematic Identification and Characterization of Novel Human Skin-Associated Genes Encoding Membrane and Secreted Proteins

    E-Print Network [OSTI]

    2013-01-01

    survey: psoriasis vulgaris (PSO), atopic dermatitis (AD),upregulated significantly in PSO, followed by AK and AD (a proinflammatory protein. PSO represents one of the major

  5. Ultraviolet a irradiation on human skin : nitric oxide mediated cardiovascular responses 

    E-Print Network [OSTI]

    Liu, Donald

    2012-11-30

    Cardiovascular disease (CVD) such as hypertension and stroke are serious illnesses that impact on the lives of millions all over the world, with 972 million (26% of the world’s population) suffering from hypertension in ...

  6. Apparatus for testing skin samples or the like

    DOE Patents [OSTI]

    Holland, J.M.

    1982-08-31

    An apparatus for testing the permeability of living skin samples has a flat base with a plurality of sample-holding cavities formed in its upper surface, the samples being placed in counterbores in the cavities with the epidermis uppermost. O-rings of Teflon washers are respectively placed on the samples and a flat cover is connected to the base to press the rings against the upper surfaces of the samples. Media to maintain tissue viability and recovery of metabolites is introduced into the lower portion of the sample-holding cavities through passages in the base. Test materials are introduced through holes in the cover plate after assembly of the chamber.

  7. Systematics of nucleon density distributions and neutron skin of nuclei

    E-Print Network [OSTI]

    Seif, W M

    2015-01-01

    Proton and neutron density profiles of 760 nuclei in the mass region of A=16-304are analyzed using the Skyrme energy density for the parameter set SLy4. Simple formulae are obtained to fit the resulting radii and diffuseness data. These formulae may be useful to estimate the values of the unmeasured radii, and especially in extrapolating charge radius values for nuclei which are far from the valley of stability or to perform analytic calculations for bound and/or scattering problems. The obtained neutron and proton root-mean-square radii and the neutron skin thicknesses are in agreement with the available experimental data.

  8. In-situ measurement of skin friction and point bearing 

    E-Print Network [OSTI]

    Rehmet, Joseph Don

    1970-01-01

    &e Dutch Eri& tion Cone. It is a device which makes separate but non- simultaneous measurement of skin friction and point bc riu;g during a si ati. c test. Objectives The objectives of this study are: 1. To design and fabricate several testing de... resistance to driving is: p P . (l + Jx) dynamic static (2) where P. , =- ma::imum dynamic rosie'lance, d~ j n a la I i (. static ma" imum st atic resist ance a viscous damping consLani used when i i. '' c i ' 'rnq i. he s Jj 1 I ' '. I . '* Lani...

  9. Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Contracts. Final report

    SciTech Connect (OSTI)

    Cornforth, Michael N.

    2013-05-03

    Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. The aims of this work apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. These aims are: to analyze by multi-flour fluorescence in situ hybridization (mFISH) the chromosomes in clonal descendents of individual human fibroblasts that were previously irradiated; to examine irradiated clones from Aim 1 for submicroscopic deletions by subjecting their DNA to comparative genomic hybridization (CGH) microarray analysis; and to flow-sort aberrant chromosomes from clones containing stable radiation-induced translocations and map the breakpoints to within an average resolution of 100 kb using the technique of 'array painting'.

  10. On the nuclear symmetry energy and the neutron skin in neutron-rich nuclei

    E-Print Network [OSTI]

    A. E. L. Dieperink; Y. Dewulf; D. Van Neck; M. Waroquier; V. Rodin

    2003-12-10

    The symmetry energy for nuclear matter and its relation to the neutron skin in finite nuclei is discussed. The symmetry energy as a function of density obtained in a self-consistent Green function approach is presented and compared to the results of other recent theoretical approaches. A partial explanation of the linear relation between the symmetry energy and the neutron skin is proposed. The potential of several experimental methods to extract the neutron skin is examined.

  11. Cysteine Dioxygenase 1 Is a Tumor Suppressor Gene Silenced by Promoter Methylation in Multiple Human Cancers

    E-Print Network [OSTI]

    2012-01-01

    Construction and Stable Cell Lines Full-length CDO1 wasof full-length CDO1 in human cancer cells markedly decreased

  12. Is the duration of skin disease visits decreasing in the united states?

    E-Print Network [OSTI]

    Davis, Scott A; Feldman, Steven R; Fleischer Jr., Alan B

    2015-01-01

    Young AC, Williford PM. Time-efficiency of nondermatologistsare generally more time-efficient in managing skin diseasein the most time-efficient manner possible. Although the

  13. An EFE model on skin-sleeve interactions during arm rotation.

    E-Print Network [OSTI]

    Xing, Malcolm M Q; Sun, Zhiguo; Pan, Ning; Zhong, Wen; Maibach, Howard I

    2006-01-01

    Sleeve Interactions During Arm Rotation Skin and garmentduring rotation of the arm. Normalized effective shearand the sleeve during the arm rotation are provided to re?

  14. Psychological Stress and skin aging: A review of possible mechanisms and potential therapies

    E-Print Network [OSTI]

    Dunn, Jeffrey H; Koo, John

    2013-01-01

    N. The grape antioxidant resveratrol for skin disorders:Therapeutic potential of resveratrol: the in vivo evidence.Kantarjian HM, et al. Resveratrol blocks interleukin-1beta-

  15. Transplantation of Adult Mouse iPS Cell-Derived Photoreceptor Precursors Restores Retinal Structure and Function in Degenerative Mice

    E-Print Network [OSTI]

    2011-01-01

    the phenomenon of HESC-derived RPE: anatomy of cell genesis,population of donor-derived photoreceptor cells (indicatedof human embryonic stem cell-derived photoreceptors restores

  16. Icariin stimulates angiogenesis by activating the MEK/ERK- and PI3K/Akt/eNOS-dependent signal pathways in human endothelial cells

    SciTech Connect (OSTI)

    Chung, Byung-Hee [Vascular System Research Center and Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chunchon, Kangwon-do 200-701 (Korea, Republic of); Division of Food Biotechnology, School of Biotechnology, Kangwon National University, Chunchon, Kangwon-do (Korea, Republic of); Kim, Jong-Dai [Division of Food Biotechnology, School of Biotechnology, Kangwon National University, Chunchon, Kangwon-do (Korea, Republic of); Kim, Chun-Ki; Kim, Jung Huan [Vascular System Research Center and Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chunchon, Kangwon-do 200-701 (Korea, Republic of); Won, Moo-Ho [Department of Anatomy, School of Medicine, Hallym University, Chunchon, Kangwon-do (Korea, Republic of); Lee, Han-Soo [Vascular System Research Center and Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chunchon, Kangwon-do 200-701 (Korea, Republic of); Dong, Mi-Sook [School of Life Sciences and Biotechnology, Korea University, 1, 5-Ka, Anam-dong, Sungbuk-ku, Seoul 136-701 (Korea, Republic of); Ha, Kwon-Soo [Vascular System Research Center and Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chunchon, Kangwon-do 200-701 (Korea, Republic of); Kwon, Young-Geun [Department of Biochemistry, College of Sciences, Yonsei University, Seoul (Korea, Republic of); Kim, Young-Myeong [Vascular System Research Center and Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chunchon, Kangwon-do 200-701 (Korea, Republic of)], E-mail: ymkim@kangwon.ac.kr

    2008-11-14

    We investigated the molecular effect and signal pathway of icariin, a major flavonoid of Epimedium koreanum Nakai, on angiogenesis. Icariin stimulated in vitro endothelial cell proliferation, migration, and tubulogenesis, which are typical phenomena of angiogenesis, as well as increased in vivo angiogenesis. Icariin activated the angiogenic signal modulators, ERK, phosphatidylinositol 3-kinase (PI3K), Akt, and endothelial nitric oxide synthase (eNOS), and increased NO production, without affecting VEGF expression, indicating that icariin may directly stimulate angiogenesis. Icariin-induced ERK activation and angiogenic events were significantly inhibited by the MEK inhibitor PD98059, without affecting Akt and eNOS phosphorylation. The PI3K inhibitor Wortmannin suppressed icariin-mediated angiogenesis and Akt and eNOS activation without affecting ERK phosphorylation. Moreover, the NOS inhibitor NMA partially reduced the angiogenic activity of icariin. These results suggest that icariin stimulated angiogenesis by activating the MEK/ERK- and PI3K/Akt/eNOS-dependent signal pathways and may be a useful drug for angiogenic therapy.

  17. Stem Cell Biology and it Application to Biotechnology

    E-Print Network [OSTI]

    Tsymbal, Evgeny Y.

    · Human embryonic stem cell research is a decade old, adult stem cell research has 30-year head start · Adult stem cells · Induced Pluripotent stem cells #12;Embryonic Stem Cells · Highest level from embryos called a Blastocyst (~5 days old, a hollow microscopic ball of cells) · are pluripotent

  18. their fate, during both normal cell turnover andafterpartof thepancreaswasremovedto

    E-Print Network [OSTI]

    Summers, Adam P.

    of removing -cells from adult human cadavers, stimulating cell replica- tion, and transplanting the resulting (ES)cells12 .Butifwecangenerate -cellsfrom adult -cells, why contend with ES cells and chromosomal aberrations. Consider, for instance, transplanting -cells from a 50- year-old cadaver

  19. Activation of ROS/NF-{kappa}B and Ca{sup 2+}/CaM kinase II are necessary for VCAM-1 induction in IL-1{beta}-treated human tracheal smooth muscle cells

    SciTech Connect (OSTI)

    Luo, S.-F. [Department of Internal Medicine, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan (China); Chang, C.-C.; Lee, I-T. [Department of Physiology and Pharmacology, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan (China); Lee, C.-W. [Department of Nursing, Division of Basic Medical Sciences, Chang Gung Institute of Technology, Chia-Yi, Taiwan (China); Lin, W.-N. [Department of Physiology and Pharmacology, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan (China); Lin, C.-C. [Department of Anesthetics, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan (China); Yang, C.-M. [Department of Physiology and Pharmacology, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan (China)], E-mail: chuenmao@mail.cgu.edu.tw

    2009-05-15

    Histone acetylation regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs) plays a critical role in the expression of inflammatory genes, such as vascular cell adhesion molecule-1 (VCAM-1). Oxidative processes have been shown to induce VCAM-1 expression. Here, we investigated the mechanisms underlying IL-1{beta}-induced VCAM-1 expression in human tracheal smooth muscle cells (HTSMCs). Our results showed that IL-1{beta} enhanced HTSMCs-monocyte adhesion through up-regulation of VCAM-1, which was inhibited by pretreatment with selective inhibitors of PKC{alpha} (Goe6976), c-Src (PP1), NADPH oxidase [diphenylene iodonium (DPI) and apocynin (APO)], intracellular calcium chelator (BAPTA/AM), PI-PLC (U73122), CaM (calmidazolium chloride), CaM kinase II (KN62), p300 (garcinol), NF-{kappa}B (Bay11-7082), HDAC (trichostatin A), and ROS scavenger [N-acetyl-L-cysteine (NAC)] or transfection with siRNAs of MyD88, PKC{alpha}, Src, p47{sup phox}, p300, and HDAC4. Moreover, IL-1{beta} stimulated NF-{kappa}B and CaMKII phosphorylation through MyD88-dependent PI-PLC/PKC{alpha}/c-Src/ROS and PI-PLC/Ca{sup 2+}/CaM pathways, respectively. Activation of NF-{kappa}B and CaMKII may eventually lead to the acetylation of histone residues and phosphorylation of histone deacetylases. These findings suggested that IL-1{beta} induced VCAM-1 expression via these multiple signaling pathways in HTSMCs. Blockade of these pathways may reduce monocyte adhesion via VCAM-1 suppression and attenuation of the inflammatory responses in airway diseases.

  20. Video Capture of Skin Motion using Calibrated Fabien DELLAS

    E-Print Network [OSTI]

    Paris-Sud XI, Université de

    3D mesh can easily be integrated and adapted in a sequence of an animated virtual human. We of virtual humans remains exhaustive and tedious. For animating virtual characters, 3D animators work similarly as drawers for car- toons, that implies hundreds of hours for only few seconds of animation

  1. Skin Thermal Injury Prediction with Strain Energy Wensheng Shen y and Jun Zhang z

    E-Print Network [OSTI]

    Zhang, Jun

    Skin Thermal Injury Prediction with Strain Energy #3; Wensheng Shen y and Jun Zhang z Laboratory, in which the activation energy includes chemical reaction only, strain energy of tissue due to thermal-dimensional model is presented for the quantitative prediction of skin injury re- sulting from certain thermal

  2. Theory of thin-skin eddy-current interaction with surface cracks N. Harfielda)

    E-Print Network [OSTI]

    Bowler, John R.

    Theory of thin-skin eddy-current interaction with surface cracks N. Harfielda) and J. R. Bowler; accepted for publication 14 July 1997 Eddy-current non-destructive evaluation is commonly performed of a typical crack. A thin-skin analysis of eddy currents is presented in which the electromagnetic fields

  3. In Vivo Biomechanics of the Fingerpad Skin Under Local Tangential Traction

    E-Print Network [OSTI]

    Hayward, Vincent

    In Vivo Biomechanics of the Fingerpad Skin Under Local Tangential Traction Qi Wang and Vincent tested in vivo for their biomechanical properties under tangential loading and for large deforma- tions words: Fingerpad skin properties; In vivo tissue measurement; Biomechanics; Fingers. Word count (main

  4. HEAT TRANSFERS IN A DOUBLE SKIN ROOF VENTILATED BY NATURAL CONVECTION IN SUMMER TIME

    E-Print Network [OSTI]

    Boyer, Edmond

    1 HEAT TRANSFERS IN A DOUBLE SKIN ROOF VENTILATED BY NATURAL CONVECTION IN SUMMER TIME P. H or in tropical and arid countries. In this work, radiation, convection and conduction heat transfers-dimensional numerical simulation of the heat transfers through the double skin reveals the most important parameters

  5. Density dependence of the symmetry energy from neutron skin thickness in finite nuclei

    SciTech Connect (OSTI)

    Vinas, X.; Centelles, M.; Roca-Maza, X.; Warda, M.

    2012-10-20

    The density dependence of the symmetry energy, characterized by the parameter L, is studied using information provided by the neutron skin thickness in finite nuclei. An estimate of L is obtained from experimental data of antiprotonic atoms. We also discuss the ability of parity violating electron scatering to obtain information about the neutron skin thickness in {sup 208}Pb.

  6. Effects of radioactive hot particles on pig skin

    SciTech Connect (OSTI)

    Kaurin, D.G.; Baum, J.W.; Schaefer, C.W. [and others

    1997-06-01

    The purpose of these studies was to determine the incidence and severity of lesions resulting from very localized deposition of dose to skin from small (< 0.5 mm) discrete radioactive particles as produced in the work environments of nuclear reactors. Hanford mini-pigs were exposed, both on a slightly off the skin, to localized replicate doses from 0.31 to 64 Gy (averaged over 1 cm{sup 2} at 70 {mu}m depth unless noted otherwise) using Sc-46, Yb-175, Tm-170, and fissioned UC{sub 2} isotopes having maximum beta-particle energies from about 0.3 to 3 MeV. Erythema and scabs (indicating ulceration) were scored for up to 71 days post-irradiation. The responses followed normal cumulative probability distributions, and therefore, no true threshold could be defined. Hence, 10 and 50% scab incidence rates were deduced using probit analyses. The lowest dose which produced 10% incidence was about 1 Gy for Yb-175 (0.5 MeV maximum energy) beta particle exposures, and about 3 to 9 Gy for other isotopes. The histopathology of lesions was determined at several doses. Single exposures to doses as large as 1,790 Gy were also given, and results were observed for up to 144 days post-exposure. Severity of detriment was estimated by analyzing the results in terms of lesion diameter, persistence, and infection. Over 1,100 sites were exposed. Only two exposed sites became infected after doses near 5000 Gy; the lesions healed quickly on treatment. 105 refs., 145 figs., 47 tabs.

  7. Enhancers and super-enhancers in human disease and therapy

    E-Print Network [OSTI]

    Hoke, Heather Ashley

    2014-01-01

    The human body is made up of a diverse array of cell types, each with specialized properties and functions that support the organism as a whole. Despite this variability, with few exceptions, these cells contain the same ...

  8. Inverse association between serum 25(OH) vitamin D levels and non-melanoma skin cancer in elderly men

    E-Print Network [OSTI]

    2010-01-01

    6):556–562 Bikle DD (2004) Vitamin D and skin cancer. J NutrSundberg JP, Welsh J (2002) Vitamin D(3) receptor ablationbetween serum 25(OH) vitamin D levels and non-melanoma skin

  9. Activated, Not Resting, Platelets Increase Leukocyte Rolling in Murine Skin Utilizing a Distinct Set of Adhesion Molecules

    E-Print Network [OSTI]

    von Andrian, Ulrich H.

    -selectin expression was studied in 8 patients with psoriasis. A correlation between platelet P-selectin expression inflammatory skin diseases. Key words: adhesion molecules/homing/inflammation/platelets/psoriasis/skin J Invest

  10. 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 [Division of Pharmacy and Toxicology, College of Pharmacy, The University of Texas at Austin, Dell Pediatric Research Institute, 1400 Barbara Jordan Blvd., Austin, TX 78723 (United States)] [Division of Pharmacy and Toxicology, College of Pharmacy, The University of Texas at Austin, Dell Pediatric Research Institute, 1400 Barbara Jordan Blvd., Austin, TX 78723 (United States); Fields, Tammy; McIvor, Elizabeth; Powell, K. Leslie; Abel, Erika L. [Department of Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Science Park, Smithville, TX 78957 (United States)] [Department of Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Science Park, Smithville, TX 78957 (United States); Vasquez, Karen M. [Division of Pharmacy and Toxicology, College of Pharmacy, The University of Texas at Austin, Dell Pediatric Research Institute, 1400 Barbara Jordan Blvd., Austin, TX 78723 (United States)] [Division of Pharmacy and Toxicology, College of Pharmacy, The University of Texas at Austin, Dell Pediatric Research Institute, 1400 Barbara Jordan Blvd., Austin, TX 78723 (United States); MacLeod, Michael C., E-mail: mcmacleod@mdanderson.org [Department of Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Science Park, Smithville, TX 78957 (United States)

    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 (?-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.

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

    SciTech Connect (OSTI)

    Gerecke, Donald R. [Environmental and Occupational Health Sciences Institute (EOHSI), a Joint Institute of UMDNJ-RW Johnson Medical School and Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ 08854 (United States)], E-mail: gerecke@eohsi.rutgers.edu; Chen Minjun; Isukapalli, Sastry S.; Gordon, Marion K.; Chang, Y.-C. [Environmental and Occupational Health Sciences Institute (EOHSI), Joint Institute of UMDNJ-RW Johnson Medical School and Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ 08854 (United States); Tong Weida [US FDA, National Center for Toxicological Research, Jefferson, AK (United States); Androulakis, Ioannis P. [Department of Biomedical Engineering, Rutgers, State University of New Jersey, Piscataway, NJ (United States); Georgopoulos, Panos G. [Environmental and Occupational Health Sciences Institute (EOHSI), Joint Institute of UMDNJ-RW Johnson Medical School and Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ 08854 (United States)

    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.

  12. Field, laboratory, and modeling investigation of the skin effect at wells with slotted casing, Boise Hydrogeophysical Research Site

    E-Print Network [OSTI]

    Barrash, Warren

    reserved. Keywords: Wellbore skin; Well hydraulics; Modeling; Pumping tests 1. Introduction Wellbore skin is a general term for imperfect hydraulic connection between a wellbore and the well structure and/or formationField, laboratory, and modeling investigation of the skin effect at wells with slotted casing

  13. Molecular Study of Interactions between Hematopoietic Stem Cells and Stromal Cells

    E-Print Network [OSTI]

    Luo, Biao

    Multipotent hematopoietic stem cells (HSCs) are progenitors of all types of hematopoietic cells, and the efficient isolation and propagation of HSCs will significantly enhance our ability to manage many human disorders ...

  14. The second skin approach : skin strain field analysis and mechanical counter pressure prototyping for advanced spacesuit design

    E-Print Network [OSTI]

    Bethke, Kristen (Kristen Ann)

    2005-01-01

    The primary aim of this thesis is to advance the theory of advanced locomotion mechanical counter pressure (MCP) spacesuits by studying the changes in the human body shape during joint motion. Two experiments take advantage ...

  15. Sensitivity of the electric dipole polarizability to the neutron skin thickness in {sup 208}Pb

    SciTech Connect (OSTI)

    Roca-Maza, X.; Agrawal, B. K.; Colo, G.; Nazarewicz, W.; Paar, N.; Piekarewicz, J.; Reinhard, P.-G.; Vretenar, D.

    2012-10-20

    The static dipole polarizability, {alpha}{sub D}, in {sup 208}Pb has been recently measured with highresolution via proton inelastic scattering at the Research Center for Nuclear Physics (RCNP) [1]. This observable is thought to be intimately connected with the neutron skin thickness, r{sub skin}, of the same nucleus and, more fundamentally, it is believed to be associated with the density dependence of the nuclear symmetry energy. The impact of r{sub skin} on {alpha}{sub D} in {sup 208}Pb is investigated and discussed on the basis of a large and representative set of relativistic and non-relativistic nuclear energy density functionals (EDF) [2].

  16. Predicting the Occurrence of Cosmetic Defects in Automotive Skin Panels

    SciTech Connect (OSTI)

    Hazra, S.; Williams, D.; Roy, R.; Aylmore, R.; Allen, M.; Hollingdale, D.

    2011-05-04

    The appearance of defects such as 'hollows' and 'shock lines' can affect the perceived quality and attractiveness of automotive skin panels. These defects are the result of the stamping process and appear as small, localized deviations from the intended styling of the panels. Despite their size, they become visually apparent after the application of paint and the perceived quality of a panel may become unacceptable. Considerable time is then dedicated to minimizing their occurrence through tool modifications. This paper will investigate the use of the wavelet transform as a tool to analyze physically measured panels. The transform has two key aspects. The first is its ability to distinguish small scale local defects from large scale styling curvature. The second is its ability to characterize the shape of a defect in terms of its wavelength and a 'correlation value'. The two features of the transform enable it to be used as a tool for locating and predicting the severity of defects. The paper will describe the transform and illustrate its application on test cases.

  17. Method for preparing dosimeter for measuring skin dose

    DOE Patents [OSTI]

    Jones, Donald E. (Idaho Falls, ID); Parker, DeRay (Idaho Falls, ID); Boren, Paul R. (Idaho Falls, ID)

    1982-01-01

    A personnel dosimeter includes a plurality of compartments containing thermoluminescent dosimeter phosphors for registering radiation dose absorbed in the wearer's sensitive skin layer and for registering more deeply penetrating radiation. Two of the phosphor compartments communicate with thin windows of different thicknesses to obtain a ratio of shallowly penetrating radiation, e.g. beta. A third phosphor is disposed within a compartment communicating with a window of substantially greater thickness than the windows of the first two compartments for estimating the more deeply penetrating radiation dose. By selecting certain phosphors that are insensitive to neutrons and by loading the holder material with neutron-absorbing elements, energetic neutron dose can be estimated separately from other radiation dose. This invention also involves a method of injection molding of dosimeter holders with thin windows of consistent thickness at the corresponding compartments of different holders. This is achieved through use of a die insert having the thin window of precision thickness in place prior to the injection molding step.

  18. Dosimeter for measuring skin dose and more deeply penetrating radiation

    DOE Patents [OSTI]

    Jones, Donald E. (Idaho Falls, ID); Parker, DeRay (Idaho Falls, ID); Boren, Paul R. (Idaho Falls, ID)

    1981-01-01

    A personnel dosimeter includes a plurality of compartments containing thermoluminescent dosimeter phosphors for registering radiation dose absorbed in the wearer's sensitive skin layer and for registering more deeply penetrating radiation. Two of the phosphor compartments communicate with thin windows of different thicknesses to obtain a ratio of shallowly penetrating radiation, e.g. beta. A third phosphor is disposed within a compartment communicating with a window of substantially greater thickness than the windows of the first two compartments for estimating the more deeply penetrating radiation dose. By selecting certain phosphors that are insensitive to neutrons and by loading the holder material with netruon-absorbing elements, energetic neutron dose can be estimated separately from other radiation dose. This invention also involves a method of injection molding of dosimeter holders with thin windows of consistent thickness at the corresponding compartments of different holders. This is achieved through use of a die insert having the thin window of precision thickness in place prior to the injection molding step.

  19. Patenting Stem Cells of Human Origin 

    E-Print Network [OSTI]

    Laurie, Graeme

    2004-01-01

    The author discusses the impact of the European Union's Directive for the legal protection of biotechnological inventions (Directive 98/44/EC of 6 July 1998. Specific attention is given to the absence from provisions of ...

  20. Human dopamine receptor and its uses

    DOE Patents [OSTI]

    Civelli, Olivier (Portland, OR); Van Tol, Hubert Henri-Marie (Toronto, CA)

    1999-01-01

    The present invention is directed toward the isolation, characterization and pharmacological use of the human D4 dopamine receptor. The nucleotide sequence of the gene corresponding to this receptor and alleleic variant thereof are provided by the invention. The invention also includes recombinant eukaryotic expression constructs capable of expressing the human D4 dopamine receptor in cultures of transformed eukaryotic cells. The invention provides cultures of transformed eukaryotic cells which synthesize the human D4 dopamine receptor, and methods for characterizing novel psychotropic compounds using such cultures.

  1. Collisional, magnetic, and nonlinear skin effect in radio-frequency plasmas Francis F. Chen

    E-Print Network [OSTI]

    Chen, Francis F.

    . CLASSICAL VS ANOMALOUS SKIN EFFECT Inductively coupled plasmas ICPs are commonly used in plasma processing.11 The experiments of Godyak et al.3­6,9 were performed with a spiral ``stove-top'' antenna, which

  2. IN FOCUS: FUTURE OF BIOSENSORS -ARTICLE Epidermal Differential Impedance Sensor for Conformal Skin

    E-Print Network [OSTI]

    Rogers, John A.

    IN FOCUS: FUTURE OF BIOSENSORS - ARTICLE Epidermal Differential Impedance Sensor for Conformal Skin requirements This article is part of the Topical Collection ``In Focus: Future of Biosensors''. X. Huang Á W

  3. Development and Construction of Bioclimatic Double Skin Active Facade for Hot and Humid Climate of UAE 

    E-Print Network [OSTI]

    Karbor, R. G.; Mohamed, I.

    2010-01-01

    become still more advanced. The present paper deals with the development and construction of mechanically ventilated double skin facade with HVAC integration for hot and humid climate like UAE. A case study is presented, illustrating potential benefits...

  4. Design Considerations for Double-Skin Facades in Hot and Humid Climates 

    E-Print Network [OSTI]

    Haase, M.; Amato, A.

    2006-01-01

    Thermal building simulations (TRNSYS) were linked to nodal airflow network simulations (COMIS) for a ventilated double-skin facade performance calculation and overall energy consumption for office building facades. Simulation results show good...

  5. Meeting report for the 1st skin microbiota workshop, Boulder, CO October 15-16 2012

    E-Print Network [OSTI]

    Gilbert, Jack A.

    This report details the outcome of the 1st Skin Microbiota Workshop, Boulder, CO, held on October 15th-16th 2012. The workshop was arranged to bring Department of Defense personnel together with experts in microbial ecology, ...

  6. A Systematic Study of Matrix Acidizing Treatments Using Skin Monitoring Method 

    E-Print Network [OSTI]

    Pandya, Nimish

    2012-07-16

    The goal of this work was to evaluate matrix acidizing treatments of vertical and horizontal wells in carbonate reservoirs. Twenty field cases for acidizing treatments were analyzed by evaluating the skin factor evolution from on-site rate/pressure...

  7. Has a thick neutron skin in ${}^{208}$Pb been ruled out?

    E-Print Network [OSTI]

    Fattoyev, F J

    2013-01-01

    The Lead Radius Experiment (PREX) has provided the first model-independent evidence in favor of a neutron-rich skin in ${}^{208}$Pb. Although the error bars are large, the reported large central value of 0.33\\,fm is particularly intriguing. To test whether such a thick neutron-skin in ${}^{208}$Pb is already incompatible with laboratory experiments or astrophysical observations, we employ relativistic models with neutron-skin thickness in ${}^{208}$Pb ranging from 0.16 to 0.33 fm to compute ground state properties of finite nuclei, their collective monopole and dipole response, and mass-{\\sl vs}-radius relations for neutron stars. No compelling reason was found to rule out models with large neutron skins in ${}^{208}$Pb from the set of observables considered in this work.

  8. REFLEXIVE COLLISION RESPONSE WITH VIRTUAL SKIN Roadmap Planning Meets Reinforcement Learning

    E-Print Network [OSTI]

    Förster, Alexander

    REFLEXIVE COLLISION RESPONSE WITH VIRTUAL SKIN Roadmap Planning Meets Reinforcement Learning Svizzera italiana, CH-6928 Manno-Lugano Keywords: Roadmap Planning: Reinforcement Learning: Collision to a changing environment, but not both. This work proposes a simple integration of roadmap planning

  9. Evaluation of the inhibition of skin matrix metalloproteinases by pothomorphe umbellata root extract

    E-Print Network [OSTI]

    Ropke, Cristina D.

    2006-10-25

    . 2003 3Ropke et al., Clin. Exp. Dermatol., 2005, 4Ropke et al., Photochem. Photobiol. 2006 in vivo results ? P. umbellata extract was able to reduce the incidence of visible and histological skin alterations in chronically UV- irradiated mice.... 2003 3Ropke et al., Clin. Exp. Dermatol., 2005, 4Ropke et al., Photochem. Photobiol. 2006 in vivo results ? P. umbellata extract was able to reduce the incidence of visible and histological skin alterations in chronically UV- irradiated mice...

  10. Oncogenic action of beta, proton, alpha and electron radiation on the rat skin

    SciTech Connect (OSTI)

    Burns, F.J.

    1980-01-01

    Rat skin is being utilized as an empirical model for testing dose and time related aspects of the oncogenic action of ionizing radiation, ultraviolet light, and polycyclic aromatic hydrocarbons. Molecular lesions in the skin DNA, including, strand breaks and thymine dimers, are being measured and compared to tumor induction. The induction and repair kinetics of molcular lesions are being compared to split dose repair. Modifiers and radiosensitizers are being utilized to test specific aspects of a chromosome breakage theory of radiation oncogenesis.

  11. Dose profiles through the dermis for on and off-skin hot particle exposures 

    E-Print Network [OSTI]

    Shaw, Kimberly Rochelle

    1993-01-01

    reports measurements of depth-dose profiles for on- and off-skin hot particle exposures using radiochromic dye film. Dose profiles from both a "Co hot particle, and activated depleted uranium oxide microspheres were measured with the film. Exposures... 24 27 32 TABLE OF CONTENTS Icontinued) Page RESULTS 34 ' Co On-Contact Exposures 34 Co Exposures Through Protective Clothing ~Co Off-Skin Exposures Uranium Microsphere On-Contact Exposures 45 49 Uranium Microsphere Exposures Through...

  12. EXPLORING THE POTENTIAL OF SHORT-TIME FOURIER TRANSFORMS FOR ANALYZING SKIN CONDUCTANCE AND PUPILLOMETRY IN REAL-TIME APPLICATIONS

    SciTech Connect (OSTI)

    Roger Lew; Brian P. Dyre; Steffen Werner; Jeffrey C. Joe; Brian Wotring; Tuan Tran

    2008-09-01

    The development of real-time predictors of mental workload is critical for the practical application of augmented cognition to human-machine systems. This paper explores a novel method based on a short-time Fourier transform (STFT) for analyzing galvanic skin conductance (SC) and pupillometry time-series data to extract estimates of mental workload with temporal bandwidth high-enough to be useful for augmented cognition applications. We tested the method in the context of a process control task based on the DURESS simulation developed by Vincente and Pawlak (1994; ported to Java by Cosentino,& Ross, 1999). SC, pupil dilation, blink rate, and visual scanning patterns were measured for four participants actively engaged in controlling the simulation. Fault events were introduced that required participants to diagnose errors and make control adjustments to keep the simulator operating within a target range. We were interested in whether the STFT of these measures would produce visible effects of the increase in mental workload and stress associated with these events. Graphical exploratory data analysis of the STFT showed visible increases in the power spectrum across a range of frequencies directly following fault events. We believe this approach shows potential as a relatively unobtrusive, low-cost, high bandwidth measure of mental workload that could be particularly useful for the application of augmented cognition to human-machine systems.

  13. Primary Radiation Therapy for Head-and-Neck Cancer in the Setting of Human Immunodeficiency Virus

    SciTech Connect (OSTI)

    Klein, Emily A.; Guiou, Michael; Farwell, D. Gregory; Luu, Quang; Lau, Derick H.; Stuart, Kerri; Vaughan, Andrew; Vijayakumar, Srinivasan; Chen, Allen M.

    2011-01-01

    Purpose: To analyze outcomes after radiation therapy for head-and-neck cancer among a cohort of patients with human immunodeficiency virus (HIV). Methods and Materials: The medical records of 12 patients with serologic evidence of HIV who subsequently underwent radiation therapy to a median dose of 68 Gy (range, 64-72 Gy) for newly diagnosed squamous cell carcinoma of the head and neck were reviewed. Six patients (50%) received concurrent chemotherapy. Intensity-modulated radiotherapy was used in 6 cases (50%). All patients had a Karnofsky performance status of 80 or 90. Nine patients (75%) were receiving antiretroviral therapies at the time of treatment, and the median CD4 count was 460 (range, 266-800). Toxicity was graded according to the Radiation Therapy Oncology Group / European Organization for the Treatment of Cancer toxicity criteria. Results: The 3-year estimates of overall survival and local-regional control were 78% and 92%, respectively. Acute Grade 3+ toxicity occurred in 7 patients (58%), the most common being confluent mucositis (5 patients) and moist skin desquamation (4 patients). Two patients experienced greater than 10% weight loss, and none experienced more than 15% weight loss from baseline. Five patients (42%) experienced treatment breaks in excess of 10 cumulative days, although none required hospitalization. There were no treatment-related fatalities. Conclusions: Radiation therapy for head-and-neck cancer seems to be relatively well tolerated among appropriately selected patients with HIV. The observed rates of toxicity were comparable to historical controls without HIV.

  14. A flow-­cytometry-­based method to simplify the analysis and quantification of protein association to chromatin in mammalian cells

    E-Print Network [OSTI]

    Forment, Josep V.; Jackson, Stephen P.

    2015-01-01

    ,  human  colon  carcinoma  HCT-­?116  cells,  human  leukaemia   HAP-­?1   cells,   human   immortalised   retinal   RPE-­?1   cells   and   primary   mouse  embryonic  fibroblasts  have  all  yielded  positive...  analysed  cell   is  also  imaged  and  processed   in   a   post-­?acquisition   manner24,25.   Alternatively,   high-­?content  quantitative  microscopy  techniques,  such  as  the  recently  described...

  15. 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.

  16. Restoration of normal phenotype in cancer cells

    DOE Patents [OSTI]

    Bissell, Mina J. (Berkeley, CA); Weaver, Valerie M. (Oakland, CA)

    1998-01-01

    A method for reversing expression of malignant phenotype in cancer cells is described. The method comprises applying .beta..sub.1 integrin function-blocking antibody to the cells. The method can be used to assess the progress of cancer therapy. Human breast epithelial cells were shown to be particularly responsive.

  17. Restoration of normal phenotype in cancer cells

    DOE Patents [OSTI]

    Bissell, M.J.; Weaver, V.M.

    1998-12-08

    A method for reversing expression of malignant phenotype in cancer cells is described. The method comprises applying {beta}{sub 1} integrin function-blocking antibody to the cells. The method can be used to assess the progress of cancer therapy. Human breast epithelial cells were shown to be particularly responsive. 14 figs.

  18. SU-E-I-53: Variation in Measurements of Breast Skin Thickness Obtained Using Different Imaging Modalities

    SciTech Connect (OSTI)

    Nguyen, U; Kumaraswamy, N; Markey, M

    2014-06-01

    Purpose: To investigate variation in measurements of breast skin thickness obtained using different imaging modalities, including mammography, computed tomography (CT), ultrasound, and magnetic resonance imaging (MRI). Methods: Breast skin thicknesses as measured by mammography, CT, ultrasound, and MRI were compared. Mammographic measurements of skin thickness were obtained from published studies that utilized standard positioning (upright) and compression. CT measurements of skin thickness were obtained from a published study of a prototype breast CT scanner in which the women were in the prone position and the breast was uncompressed. Dermatological ultrasound exams of the breast skin were conducted at our institution, with the subjects in the upright position and the breast uncompressed. Breast skin thickness was calculated from breast MRI exams at our institution, with the patient in the prone position and the breast uncompressed. Results: T tests for independent samples demonstrated significant differences in the mean breast skin thickness as measured by different imaging modalities. Repeated measures ANOVA revealed significant differences in breast skin thickness across different quadrants of the breast for some modalities. Conclusion: The measurement of breast skin thickness is significantly different across different imaging modalities. Differences in the amount of compression and differences in patient positioning are possible reasons why measurements of breast skin thickness vary by modality.

  19. Annexin A2 Modulates Radiation-Sensitive Transcriptional Programming and Cell Fate

    SciTech Connect (OSTI)

    Waters, Katrina M.; Stenoien, David L.; Sowa, Marianne B.; Freiin Von Neubeck, Claere H.; Chrisler, William B.; Tan, Ruimin; Sontag, Ryan L.; Weber, Thomas J.

    2012-11-13

    There is considerable public interest in the health effects of low doses of radiation (LDR) that fall below the doses that can be plausibly investigated in epidemiological studies. At these low doses, experimental models can detect perturbations in signaling pathways and use this information to define functional consequences of LDR exposures prospectively. In this study, we show increased nuclear annexin A2 (AnxA2) levels in human skin organotypic culture and murine progenitor cell model systems following exposure to X-radiation (10-200 cGy). LDR (2-20 cGy) inhibits cell transformation responses following epidermal growth factor (EGF) or 12-O-tetradecanoylphorbol-13-acetate (TPA) exposures, indicating LDR may have a protective component mediated in part by nuclear localization of AnxA2. Oncogenic protein kinase C epsilon (PKC?) levels are increased in nuclear extracts from AnxA2 silenced [shRNA] cells, suggesting that AnxA2 may contribute to PKC? nuclear export, perhaps reducing oncogenic potential. Coordinately, silencing AnxA2 results in a sensitive phenotype and cells grow constitutively in soft agar. Using global microarray analysis, we show that silencing AnxA2 fundamentally alters transcriptional programming, changing the radioresponsive transcriptome and revealing biological processes that are induced in the absence of AnxA2. These observations suggest that AnxA2 plays a fundamental role in the sensitivity of cellular and tissue response to ionizing radiation, and deficiency of AnxA2 could result in a permissive environment for radiation-induced health effects.

  20. A basal stem cell signature identifies aggressive prostate cancer phenotypes.

    E-Print Network [OSTI]

    2015-01-01

    RSEM). Comparing CD49f Hi Gene Signature to Other Stem Cellcharacterize the Trop2 + CD49f Hi human basal stem cellidentified Trop2 + CD49f Hi human basal stem cell population

  1. The Effect of the iBEAM Evo Carbon Fiber Tabletop on Skin Sparing

    SciTech Connect (OSTI)

    Simpson, John B. Godwin, Guy A.

    2011-10-01

    Replicating the attenuation properties of the treatment tabletop are of primary importance for accurate treatment planning; however, the effect of the tabletop on the skin-sparing properties of x-rays can be overlooked. Under some conditions, the reaction of skin to the radiation can be so serious as to be the dose-limiting organ for radiotherapy treatment. Hence, an understanding of the magnitude of the reduction in skin sparing is important. Because of the development of image-guided radiotherapy, modern tabletops have been developed without the use of metal supports that otherwise provided the necessary level of rigidity. Rigidity is instead provided by compressed foam within a carbon-fiber shell, which, although it provides artefact-free imaging and high levels of rigidity, has an adverse affect on the dose in the build-up region. Representative of this type is the iBEAM evo tabletop, whose effect on the skin dose was determined at 6-MV, 10-MV, and 18-MV x-rays. Skin dose was found to increase by 60-70% owing to the tabletop, with the effect increasing with field size and decreasing with energy. By considering an endpoint of erythema, a radiobiological advantage of selecting 10 MV over 6 MV for applicable treatments was demonstrated.

  2. Human Subjects Section 6. Protection of Human

    E-Print Network [OSTI]

    Heller, Barbara

    Human Subjects Section 6. Protection of Human Subjects This section is required for applicants answering "yes" to the question "Are human subjects involved?" on the R&R Other Project Information form subjects applicants must provide a justification in this section for the claim that no human subjects

  3. Detecting and molecular profiling cancer cells in patients

    E-Print Network [OSTI]

    Peterson, Vanessa M. (Vanessa Marie)

    2013-01-01

    Although tumor cells obtained from human patients by surgical biopsy, image-guided intervention, blood draws or fluid drainage (paracentesis, thoracentesis) are a valuable source for analyzing tumor cells, conventional ...

  4. HER receptor-mediated dynamic signalling in breast cancer cells 

    E-Print Network [OSTI]

    Hu, Huizhong

    2011-07-05

    The dynamics of cell signalling are critical to cell fate decisions. Human Epidermal growth factor Receptors (HERs)-mediated Ras/Raf/MEK/ERK and PI3K/Akt signalling cascades relay extracellular signals from the plasma ...

  5. In vitro models for airway epithelial cell culture

    E-Print Network [OSTI]

    Sivathanu, Vivek

    2013-01-01

    This work is about the development of a physiologically relevant model of the human airway. Various factors such as the cell model, physiochemical factors such as the cell substrate properties including its stiffness, shear ...

  6. NF-kappa B- and AP-1-mediated induction of human beta defensin-2 in intestinal epithelial cells by Escherichia coli Nissle 1917: A novel effect of a probiotic bacterium

    E-Print Network [OSTI]

    2004-01-01

    and E. Isolauri. 2002. Probiotics: an overview of bene?cial17. Isolauri, E. 2001. Probiotics in human disease. Am. J.and S. Salminen. 2002. Probiotics: a role in the treatment

  7. Multi-scale imaging and informatics pipeline for in situ pluripotent stem cell analysis

    E-Print Network [OSTI]

    Gorman, Bryan Robert

    2015-01-01

    Human pluripotent stem (hPS) cells have the ability to reproduce indefinitely and differentiate into any cell type of the body, making them a potential source of cells for medical therapy and an ideal system to study fate ...

  8. Generation of Isogenic Pluripotent Stem Cells Differing Exclusively at Two Early Onset Parkinson Point Mutations

    E-Print Network [OSTI]

    Soldner, Frank

    Patient-specific induced pluripotent stem cells (iPSCs) derived from somatic cells provide a unique tool for the study of human disease, as well as a promising source for cell replacement therapies. One crucial limitation ...

  9. Cell Stem Cell Stem Cell States, Fates,

    E-Print Network [OSTI]

    Peterson, Carsten

    Cell Stem Cell Review Stem Cell States, Fates, and the Rules of Attraction Tariq Enver,1 Martin Pera,2 Carsten Peterson,3,4 and Peter W. Andrews5,* 1The Molecular Haematology Unit, The Weatherall and their relationship to commitment to differ- entiate and lineage selection can be elucidated in terms of a landscape

  10. Total skin electron beam therapy using an inclinable couch on motorized table and a compensating filter

    SciTech Connect (OSTI)

    Fuse, H.; Suzuki, K.; Shida, K.; Takahashi, H.; Kobayashi, D.; Seki, M.; Mori, Y.; Sakae, T.; Isobe, T.; Okumura, T.; Sakurai, H.

    2014-06-15

    Total skin electron beam is a specialized technique that involves irradiating the entire skin from the skin surface to only a few millimetres in depth. In the Stanford technique, the patient is in a standing position and six different directional positions are used during treatment. Our technique uses large electron beams in six directions with an inclinable couch on motorized table and a compensating filter was also used to spread the electron beam and move its intensity peak. Dose uniformity measurements were performed using Gafchromic films which indicated that the surface dose was 2.04 ± 0.05 Gy. This technique can ensure the dose reproducibility because the patient is fixed in place using an inclinable couch on a motorized table.

  11. Adapting Induced Pluripotent Stem Cells For Clinical Use

    E-Print Network [OSTI]

    Brutlag, Doug

    ://www.nature.com/nbt/journal/v27/n5/full/nbt.1535.html · Kim, D. et al. Generation of human induced pluripotent stem cellsAdapting Induced Pluripotent Stem Cells For Clinical Use Dana Wyman Professor Brutlag Genomics Pluripotent Stem Cells 7 days hES medium +MEF #12;Immunofluorescence Test ES Cells (+ control) Untransduced

  12. Chronic cellular responses of rat skin to 13 Mev proton irradiation 

    E-Print Network [OSTI]

    Hinkle, Donald King

    1966-01-01

    irradiated in a total of six rad groups as follows: Number in Grou dD 10 6 9 6 7 8 ZOO 400 700 1300 2000 2500 All sections of skin and tumor tissues were submitted to the Anatomic Pathology Section, USAF School of Aerospace Medicine...CHRONIC CELLULAR RESPONSES OF RAT SKIN TO 13 MEV PROTON IRRADIATION A Thesis by DONALD KING HINKLE, D. V. M. Submitted to the Graduate College of the Texas AErM University in partial fulfillment of the requirements for the degree of MASTER...

  13. Human retinoblastoma gene

    SciTech Connect (OSTI)

    Bookstein, R.; Lee, E.Y.H.P.; Peccei, A.; Lee, W.H. (Dept. of Pathology M-012 and Center for Molecular Genetics, Univ. of California, San Diego, La Jolla, CA (US))

    1989-04-01

    Mutational inactivation of the retinoblastoma (RB) gene is considered a fundamental event in the formation of several types of human cancer. A substantial proportion of RB gene mutations are partial or complete deletions that extend an unknown distance beyond one or both ends of the gene. To provide a framework for measuring the extent of these deletions, the authors have constructed a long-range restriction map of SfiI sites spanning 850 kilobases around the RB gene. This map was applied in a molecular analysis of RB gene deletion in breast cancer cell line MB468. A previous study of this cell line demonstrated deletion of the entire RB gene except for exons 1 and 2. Genomic clones containing the deletion junction were isolated from a library made from MB468 DNA. A probe obtained from the far side of the deletion junction was used to localize and clone the unknown 3' endpoint, demonstrating that the chromosomal mutation in this case was a simple deletion spanning 200 kilobases. Sequence analysis of the deletion junction indicated a conservative deletion with no loss or gain of nucleotides. The deletion endpoints had no sequence homology to each other or to any repetitive sequence family, such as Alu, so the recombination event was illegitimate. Structural analysis of this and other RB gene deletions is important for understanding molecular mechanisms of recessive oncogenesis.

  14. Core-Protected Platinum Monolayer Shell High-Stability Electrocatalysts for Fuel-Cell Cathodes

    SciTech Connect (OSTI)

    Adzic, R.R.; Sasaki, K.; Naohara, H.; Cai, Y.; Choi, Y.M.; Liu, P.; Vukmirovic, M.B.; Wang, J.X.

    2010-11-08

    More than skin deep: Platinum monolayers can act as shells for palladium nanoparticles to lead to electrocatalysts with high activities and an ultralow platinum content, but high platinum utilization. The stability derives from the core protecting the shell from dissolution. In fuel-cell tests, no loss of platinum was observed in 200?000 potential cycles, whereas loss of palladium was significant.

  15. P4.37 INTER-COMPARISON OF GOES-8 IMAGER AND SOUNDER SKIN TEMPERATURE RETRIEVALS

    E-Print Network [OSTI]

    Haines, Stephanie L.

    * University of Alabama in Huntsville Global Hydrology and Climate Center, Huntsville, AL Ronnie J. Suggs and Gary J. Jedlovec Global Hydrology and Climate Center MSFC/NASA, Huntsville, AL 1. INTRODUCTION Skin (GOES) data at the Global Hydrology and Climate Center (GHCC). The GOES Imager or Sounder data are used

  16. Characterization of Urinary Iron Loss in the fsn (flaky skin) Anemia Mouse Mutant

    E-Print Network [OSTI]

    Kress, Robert Lee

    2014-08-31

    target to treat iron overload. The flaky–skin anemia (fsn) mouse possesses a mutation in the Ttc7 gene (tetratricopeptide repeat domain 7) and had been observed to excrete elevated iron levels in its urine. We hypothesized that the mutation in fsn...

  17. ON THE INFLUENCE OF THE GEOMETRY ON SKIN EFFECT IN ELECTROMAGNETISM

    E-Print Network [OSTI]

    Faou, Erwan

    ON THE INFLUENCE OF THE GEOMETRY ON SKIN EFFECT IN ELECTROMAGNETISM GABRIEL CALOZ, MONIQUE DAUGE, ERWAN FAOU, VICTOR P´ERON ABSTRACT. We consider the equations of electromagnetism set on a domain made in electromagnetism. This effect describes the rapid decay of electromagnetic fields with depth inside a metallic

  18. Vessel Segmentation and Analysis in Laboratory Skin Transplant Micro-angiograms

    E-Print Network [OSTI]

    Lübeck, Universität zu

    Vessel Segmentation and Analysis in Laboratory Skin Transplant Micro-angiograms Alexandru transplantations depends on the adequate revascularization of the trans- planted dermal matrix. To induce vessel and length of newly grown vessels have to be measured in micro-angiograms (x-ray images of the blood vessels

  19. Water skin anomalies: density, elasticity, hydrophobicity, thermal stability, interface repulsivity, etc

    E-Print Network [OSTI]

    Chang Q. Sun

    2015-02-26

    Molecular undercoordination induced O:H-O bond relaxation and dual polarization dictates the supersolid behavior of water skins interacting with other substances such as flowing in nanochannels, dancing of water droplets, floating of insects. The BOLS-NEP notion unifies the Wenzel-Cassie-Baxter models and explains controllable transition between hydrophobicity and hydrophilicity.

  20. Experimental evaluation of a naturally ventilated PV double-skin building envelope in real operating conditions

    E-Print Network [OSTI]

    Paris-Sud XI, Université de

    . Introduction France is undergoing an energy transition towards technologies with a lower environmental impact Fax. +33472438811 Abstract Building integrated photovoltaic systems are fast becoming a feature of a prototype naturally-ventilated photovoltaic double-skin facade, designed to maintain favourable operating