Accessibility of epitopes on UvrB protein in intermediates generated during incision of UV-irradiated DNA by the Escherichia coli Uvr(A)BC endonuclease

Structural intermediates generated during incision of damaged DNA by the Uvr(A)BC endonuclease were probed with monoclonal antibodies (mAbs) raised against the Escherichia coli UvrB protein. It was found that the epitope of B2C5 mAb, mapped at amino acids (aa) 171-278 of UvrB, is not accessible in any of the preformed Uvr intermediates. Preformed B2C5-UvrB immunocomplexes, however, inhibited formation of those intermediates. B2C5 mAb seems to interfere with the formation of the UvrA-UvrB complex due to overlapping of its epitope and the UvrA binding region of UvrB. Conversely, the epitope of B3C1 mAb (aa 1-7 and/or 62-170) was accessible in all Uvr intermediates. The epitope of B*2E3 mAb (aa 171-278) was not accessible in any of the nucleoprotein intermediates preceding UvrB-DNA preincision complex. However, B*2E3 was able to immunoprecipitate this complex and to inhibit overall incision. B2A1 mAb (aa 8-61) inhibited formation of those Uvr intermediates requiring ATP binding and/or hydrolysis by UvrB. B*2B9 mAb (aa 473-630) inhibited Uvr nucleoprotein complexes involving UvrB. B*2B9 seems to prevent the binding of the UvrA-UvrB complex to DNA. The epitope of the B*3E11 mAb (aa 379-472) was not accessible in Uvr complexes formed at damaged sites. These results are discussed in terms of structure-functional mapping of UvrB protein.     

Structural changes in membranes of proliferating leukocytes L-41 as affected by low doses of ionizing radiation

The dose-dependent effects of gamma-radiation on the leucocyte cultures L-41 has been investigated. Irradiation by the dose of 0.25 Gy stimulates the cell proliferation while that by the doses of 1.0 and 2.0 Gy inhibits this process. In this dose range the radiohormesis effects characterizing structural organization of the probed membranes have been also registered for fluorescence parameters. The zone of qualitative transition of response of the cell membranes to radiation is individual for various effects. The action of radiation in the doses of 0.25 and 0.50 Gy induces a decrease of ANS fluorescence intensity and a decrease of membrane protein immersion in the lipid bilayer of leukocyte membranes. The values of these parameters rise at the doses of 1.0 and 2.0 Gy that reflects different directions of structural changes in various membrane regions. The irradiation in the range of 0.25-1.0 Gy induces the increase of microviscosity in deep regions of membrane lipid matrix while at the dose 2.0 Gy it causes its decrease. Radioactive radiation does not change the membrane protein conformation of leukocytes and polarity of lipid bilayer hydrophobic zones as recorded by fluorescent methods used.     

Use of conduction anesthesia for the development of movements in the knee joint after arthrotomy

An investigation of the possible significance of 'stress' as a factor in the retention of bovine fetal membranes has been carried out, together with the investigation as to the possible variation of progesterone levels at parturition, as a cause of retention. No evidence of abnormal progesterone levels has been established, but from the evidence it is concluded that retention may well be yet another example of production disease.     

Effects of the synthetic anti-oxidant, probucol, on the U937 monoblastoid cell line

We examined physiological events in the hyperplastic artery, using a method based on the mechanical responsiveness and myosin light-chain phosphorylation in response to various stimulants. Six weeks after endothelial denudation by ballooning of the right carotid artery, strips of this artery with moderate intimal hyperplasia (intimal area was 30-50% of medial area in 20 of 28 rabbits) were used for experiments. Strips from the left carotid served as the normal control. When the hyperplastic artery was stimulated with 30 microM PGF2 alpha, the maximal tension (232.4 +/- 49.1 mg/mg dry wt, mean +/- SD) was significantly higher (P < 0.05) than that of the control (129.5 +/- 16.4 mg/mg). The maximal extent of myosin light-chain monophosphorylation (45.4 +/- 8.9%) and diphosphorylation (10.9 +/- 5.2%) in the hyperplastic artery was significantly higher (P < 0.05) than that in the control artery (33.0 +/- 4.8 and 4.0 +/- 4.8%, respectively). The monophosphorylation of the myosin light chain in the hyperplastic artery was sustained for up to 20 min, while that in the control artery decreased to the basal level within 20 min. Similar observations were obtained by stimulation with 60 mM K+ or 30 microM norepinephrine. Dose-response curves of the development of tension in the hyperplastic artery to various agonists (K+, PGF2 alpha, norepinephrine) shifted upward the curves for the control artery. These results suggest that qualitative changes in the characteristics of smooth muscle cells may occur in the intimal hyperplastic portion, including a hyperreactive contraction associated with enhanced and sustained phosphorylation of the myosin light chain.