Procedures for the assay of (2'-5')-oligoadenylic acids and their derivatives in intact cells.
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Biomedical subjects
Publications and source records attributed to B R Williams.
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Successful control of diabetes mellitus requires that the patient assume primary responsibility for treatment of his or her disease. The University of Southern California Schools of Medicine and Pharmacy, using the Pico Rivera Health Center as a teaching site, instituted an educational program for diabetic members of the community. Pretests and posttests showed that there was an increase in knowledge for those who participated in the classes. The pharmacist and physician interns also gained experience in providing community education.
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Cells cultured from calf ligamentum nuchae produce extracellular microfibrils identical to those of intact elastic tissue as determined by ultrastructural appearance, degradative enzyme susceptibilities and amino acid composition. A method to isolate large amounts of the microfibrillar component using fluorescein mercuric acetate is described. The cultures do not appear to synthesize the amorphous component, elastin, in that radioactive desmosine and isodesmosine were not detected after incubating cultures with labeled lysine nor was elastin seen ultrastructurally.
Until now the interferon-mediated 2'-5' adenine oligonucleotide inhibitors (2-5A) of cell-free protein synthesis have not been detected in intact cells. Here we report their natural occurrence in interferon-treated, EMC virus-infected mouse L cells in amounts consistent with the idea that they play a part in the inhibition of virus growth.
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Double-stranded RNA (dsRNA) inhibits protein synthesis in rabbit reticulocyte lysates by activating the synthesis of the endonuclease effector pppA2' p5' A2' p5' A(2-5A) and a protein kinase which phosphorylates the protein synthesis initiation factor eIF-2. Under certain assay conditions, high concentrations of dsRNA are without inhibitory effect in many lysates (high dsRNA "reversible" lysates). In these lysates natural dsRNA at low concentrations stimulated protein kinase activity to a greater extent than did the synthetic dsRNA poly rI.rC. Synthesis of 2--5A was greater when poly rI.rC was used. However, a number of factors, including the salt concentration and messenger RNA used, combine to determine the overall effect of dsRNA on protein synthesis under any given set of experimental conditions.
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The effects of heat treatment on Blue Elgiloy wire and loops formed from this wire were determined. The optimum heat treatment temperature was established to be in a narrow range in the vicinity of 950 degrees F. This regimen increases the yield and ultimate strengths of this wire. Heat treatment also produced a modification of the load-deflection characteristics of the loop configuration tested. Overall, heat treatment of the wire is indicated by this study.
Incubation of the mouse L-cell-free system with a concentration of pppA2'p5'A2'p5'A [(2'-5')An] just sufficient to inhibit protein synthesis results in formation of a high-molecular-weight, heatlabile inhibitor and enhanced ribonuclease activity and in the rapid breakdown of (2'-5')An to ATP. The (2'-5')An-enhanced ribonuclease activity is also unstable and in the absence of a (2'-5')-An-regenerating system inhibiton of protein synthesis is transient. Although interferon treatment enhances the synthesis of (2'-5')An, the rates of degradation of (2'-5')An and levels of activatible nuclease are similar in extracts prepared from control or interferon-treated cells. Interestingly, the sensitivity of different cell-free systems to (2'-5')An, varies with the source of the cell-free systems and with the methods used in their preparation. There is, however, no obvious correlation between the sensitivities of the system and the rate of breakdown of (2'-5')An. The significance of these results is discussed in relation to a possible control function for the (2'-5')An system in both interferon-treated and control cells.
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