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STUDY OF THE HEALING OF CORNEAL WOUNDS WITH RADIOACTIVE ISOTOPES, WITH SPECIAL REFERENCE TO THE PLASTIC ARTIFICIAL CORNEA. Final Report

Technical Report ·
OSTI ID:4094190

The ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) content of corneal epithelium and corneal stroma were measured, the RNA was separated into its component nucleotides, and the uptake of P/sup 32/ (phosphate) into the nucleic acids was determined. Phosphorus-containing compounds of rabbit, calf, and dogfish corneas were extracted by a modification of the Schmidt-Thannhauser procedure. Hot 10% NaCl was then used to extract the nucleic acids from the defatted and acid-precipitated tissues. After the RNA and DNA were precipitated by cold 95% ethanol and separated by hydrolyzing the RNA with dilute KOH, the products of this extraction were lyophylized and ultraviolet absorption studies, phosphorus analyses, and sugar tests were performed. For the epithelial tissue, sugar tests and ultraviolet absorption spectra showed characteristics of RNA and DNA; no such characteristics emerged for stromal tissue. A system was developed whereby labeled nucleic acid precursors were incorporated into the RNA and DNA of incubated corneas. Studies of the in vitro uptake of P/sup 32/ (phosphate) by the cornea revealed that epithelial RNA and DNA fractions contained radioactivity after incubation. Radioactivity was detected in the stroma, but the P/sup 32/ (phosphate) could not be identified as RNA or DNA. Paperelectrophoretic separations of corneal epithelial ribonucleotides showed that P/sup 32/ (phosphate) was incorporated into all four nucleotides during a two-hour incubation in KrebsRinger-bicarbonate medium. The relative amounts, specific activities, and nucleotide ratios of nucleic acids present differentially in epithelium and stroma of calf, rabbit, and elasmobranch normal corneas are shown. Results indicate that in the normal cornea at least, the epithelium has little effect on the nucleic acid or the phosphorus metabolism of the stroma. (auth)

Research Organization:
Harvard Univ., Boston. Massachusetts Eye and Ear Infirmary
NSA Number:
NSA-18-013418
OSTI ID:
4094190
Report Number(s):
TID-13636
Country of Publication:
United States
Language:
English