Use of spin-labelled substrate analogs for a comparative study of microsomal cytochrome P-450 and P-448

The previously described, iodine-labeled alkylating stable nitroxyl radicals located at different distances between the N-O. group and the iodine atom were used for a comparative study of the structure of microsomal cytochromes P-450 and P-448 active centers. The radicals were shown to change the optical spectra of Fe3+ located in the active site of the enzyme that are similar to those induced by cytochrome P-450 substrates. Some differences in the type of the radicals binding to control, phenobarbital- and 3-methylcholanthrene-induced microsomes were revealed. The alkylating radical substrate analogs covalently bound to microsomal cytochrome P-450 in the vicinity of the active center, resulting in the inhibition of oxidation of type I and II substrates (e. g., aniline and naphthalene). The value of the spectral binding constant (Ks) for naphthalene in the presence of the radical covalently bound to the cytochrome P-450 active center showed a tendency to increase. Using the ESR technique, the interaction between Fe3+ and the radical localized in the active site of cytochrome P-450 was demonstrated. The contribution of Fe3+ to the relaxation of the radicals covalently bound to cytochrome P-450 was evaluated from the values of the spin label ESR spectra saturation curves at 77K. The distances between the N-O. group of these radicals and Fe3+ in the enzyme active center for the three types of microsomes were determined. The data obtained point to structural peculiarities of the active center of cytochrome P-450, depending on the microsomal type.     

Effect of adenine nucleotides on the activator function of streptokinase

The addition of ATP or 3,5-AMP (but not UTP, GTP, CTP, AMP, 2,3-AMP, ADP, inorganic pyrophosphate) at a final concentration of 10(-1) M into streptokinase solution, pH 7.0 or 9.5, causes a dramatic inhibition of streptokinase-induced fibrinolysis. The specificity of ATP effect is fully lost at pH 3.0, when all nucleotides completely inhibit the activating function of streptokinase. Ribose-5-phosphate causes a similar effect at pH 3.0. The character of nucleotide action on the activating function of streptokinase considerably differs from their influence on proteolytic reactions.     

Mitogenic action of bacterial T-independent antigens under various conditions of lymphocyte culture

The mitogenic response of lymphocytes in mouse spleen cell culture to the action of Salmonella typhi lipopolysaccharide (LPS) and Vi-antigen under different experimental conditions has been studied. The density of the culture has been shown to influence the activation of lymphocytes with Vi-antigen. Thus, macrophages stimulate mitogenesis at the concentration of lymphocytes equal to 1.0-1.2 X 10(6) cells/ml and suppress it when this concentration increases tenfold. The method used for the purification of cell suspension from adhering cells has been shown to influence the level of the mitogenic response of lymphocytes. The cultures, purified from macrophages by filtration through a column packed with cotton wool, have been found to respond to the mitogenic doses of LPS 7-8 times weaker than those purified by adhesion onto plastic. Background and mitogen-induced inclusion of 3H-thymidine into lymphocytic DNA varies in accordance with the presence of adhering cells. For this reason, in the evaluation of the influence of macrophages on mitogenesis it is expedient to consider not only the stimulation index, but also the absolute inclusion of thymidine into cells.     

Genetics of congenital esophageal defects

An analysis of 304 cases of esophageal atresia in fetuses and neonates showed that frequency of other congenital malformations is 43.1% including 10% of chromosomal disorders and monogenous syndromes. Summarizing the authors' own data and evidences from literature the genetic risk for sibs is calculated to be 0.88% and heredity--57.3 +/- 5.1%. The hypothesis that esophageal atresia is a malformation of multifactorial genesis with polygenic hereditary component is confirmed.     

Purification, quaternary structure and immunological properties of phosphorylase kinase from chicken skeletal muscle

Phosphorylase kinase was isolated from red and white chicken skeletal muscle in a nearly homogeneous state as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The molecular weight of the native enzyme as determined by gel filtration on Sepharose 4B is close to that of rabbit skeletal muscle phosphorylase kinase (i. e., approximately 1300 000). The molecular weights of the subunits determined by SDS gel electrophoresis are: alpha', 140 000 beta, 129 000; gamma', 44 000; delta, 17 000 (cf. the Mr values of the alpha- and gamma-subunits of the rabbit muscle isoenzyme are 146 000 and 42 000). The four subunits, alpha', beta, gamma' and delta, were found to exist in equimolar amounts as shown by a densitometric analysis of acrylamide gels; hence, the subunit formula of the chicken skeletal muscle isoenzyme is (alpha' beta gamma' delta)4. Rabbit antisera against a mixture of alpha'- and beta-subunits of chicken phosphorylase kinase yield a single precipitin line with this enzyme, do not show cross reactions of identity with the rabbit muscle enzyme but strongly inhibit the activity of the chicken enzyme and partially inhibit the activity of the rabbit muscle isoenzyme.     

Ascorbate-independent carotenoid de-epoxidation in intact spinach chloroplasts

Slow (> 1 s) light-induced absorbance changes in the 475–530 nm spectral region were examined in Type A chloroplasts from spinach. The most prominent absorption change occurred at 505 nm. The difference spectrum for this light-induced increase, its absence in osmotically shocked chloroplasts and restoration by ascorbate, and its sensitivity to dithiothreitol indicate that the absorption change is due to carotenoid de-epoxidation. The reaction in intact chloroplasts is characterized by its independence of exogenous ascorbate and a rate constant 3- to 8-fold higher than that reported previously for chloroplasts supplemented with ascorbate.The relevance of carotenoid de-epoxidation to other photosynthetic processes was examined by comparing their sensitivities to dithiothreitol. Levels of dithiothreitol that eliminate the 505 nm shift are without effect on saturated rates of CO₂ fixation and do not appreciably inhibit fluorescence quenching. We conclude that carotenoid de-epoxidation is not directly involved in the reactions of photosynthesis or in the regulation of excitation allocation between the photosystems.     

Experimental effect of ultraviolet laser radiation on lung, tracheal and bronchial tissues

The authors studied morphological changes under the effect of laser radiation. Various disorders in the blood circulation have been revealed such as hyperemia, stasis which did not depend upon the exposition time.     

Early Siberians from Lake Baikal and Alaskan population affinities

Among the materials excavated by the 1975 joint USSR–USA team in Siberia are two burials from Shaman's Cape, Olkhon Island, Lake Baikal. One is a middle-aged male of the Serovo culture, 6,000 B.C. , and the other is a young male of the Glaskovo culture of 2,000 B.C. This later burial displays an unusual pathology affecting the nose and post-cranial regions of the pelvic girdle and lower limbs. Osteon analysis confirms the determination of age at death and illustrates the difference between normal and pathological bone. Numerous cultural materials were associated with these burials, including harpoon heads, knives, a compound fishhook and a pestle with the Serovo man, and nephrite ornaments with the Glaskovo man. The skulls, though far apart in time, are pronouncedly Mongoloid and alike in their low cranial vaults. A low, broad and inclined ascending ramus resembles Chukchi, Eskimos and Aleuts. These two specimens document the Mongoloid character of the early inhabitants of Lake Baikal.     

Secretion of organic acids and bases by the kidneys of marine teleosts

In the experiments performed on kidneys of 5 species of marine Teleostei, morphological peculiarities in secretion of 8 fluorescent organic acids (uranin, primulin, tripaphlavin, erythrosin etc.) and in 5 organic bases (rhodamin C, auramin etc.) have been studied. At a very low concentration in the incubation medium--about 0.005 mg/ml--the substances mentioned penetrate into the epithelial cell of the canaliculus; its weak fluorescence appears, and soon they begin to be excreted in great amount through the apical part of the plasmolemma and accumulated in the canalicular lumen. All the substances studied accumulate in the cell and only some of them (uranin, primulin, titanic yellow, etc.) are secreted into the canalicular lumen. Penicillinum and probenecide inhibit penetration of the organic acids into the cell through the basal membrane. Uranin secretion into the canalicular lumen is inhibited in the presence of furocemid; amilorid, magnium and sulfate ions do not influence secretion of the organic acids. Secretion of the organic bases does not change when paraaminohippuric acid and furocemid are added to the medium, but it decreases when concentration of magnium ions increases.