Mutations of the VCORC1 Gene of Resistance to Anticoagulants in Synanthropic Rodents in Urbanized Areas of Russia.

This is the first study to analyze the variability of two exons (the 1st and 3rd) of the VCORC1 gene in Russian populations of mice and rats, namely, 125 Mus musculus and 19 Rattus norvegicus captured in 13 settlements. Previously, it has been shown that a number of mutations in these exons in the countries of Western Europe are associated with resistance to anticoagulants in synanthropic rodents. We did not find such mutations in house mice in Russia. However, in the first exon, two previously unknown mutations were identified that could potentially have such an effect (Lys58Arg and Ser31Trp). In three districts of Moscow, Norway rats were found carrying in the third exon a previously known resistance mutation (Tyr139Ser) in the heterozygous state. The results are discussed in connection with the intensity of the use of anticoagulants in the settlements of Russia and the mutation rate of the VCORC1 gene.

[Toxic and genetic risk assessments in children of Gornaya Shoria].

This comprehensive study assessed a number of environmental factors that were potentially able to induce genotoxic effects in man. A set of radiological, physicochemical, and bioindication techniques was used to estimate the quality of water, air, and soil in the places of residence and education of children and adolescents from the boarding school of the town of Tashtagol and schoolchildren from the village of Krasnoye, Kemerovo Region. Excess radon levels in the air of living spaces and classes, a small excess of the maximum allowable concentration of gross forms of heavy metals in individual soil samples, and high toxic effects in the Drosophila gametes exposed to air samples were revealed in the Tashtagol children having higher genotoxic effects in the lymphocyte than those in the Krasnoye village ones. The findings suggest that the marked genotoxic effects recorded over a long time in the lymphocytes of children and adolescents living in Tashtagol may be associated with the complex influence of the above factors. It is north supplementing further investigations by an extended chemical analysis of water, air, snow, and soil samples taken in the places of residence and education of children.

Conjugation of the Bowman-Birk soybean proteinase inhibitor with hydroxyethylstarch.

The classical Bowman-Birk soybean proteinase inhibitor was modified by hydroxyethylstarch. The modified inhibitor retained the capacity for simultaneous binding of trypsin and human leukocyte elastase. The inhibition constants, Ki, of bovine trypsin, alpha-chymotrypsin and human leukocyte elastase (HLE) increased not more than 10-, 1.5-, and 20-fold, respectively, on modification of the inhibitor. The less effective inhibition is presumably due to the steric hindrance brought about by the conjugation with polysaccharide molecules. The results obtained indicate the pronounced structure differences of the binding regions for trypsin and chymotrypsin/leukocyte elastase in the modified preparation.

[Development of a method of polarization fluorescence immunoanalysis of the herbicide 2,4,5-trichlorophenoxyacetic acid]. / Razrabotka poliarizatsionnogo fliuorestsentnogo immunoanaliza gerbitsida 2,4,5-trikhlorfenoksiuksusnoi kisloty.

Immunoreagents were produced and a polarographic fluorescent immunoassay was developed for estimation of residual content of widely used herbicide 2,4,5-trichlorophenoxy acetic acid. The procedure is simple, accurate, specific and allowed one to measure 10 preparations within 7 min. The sample volume was 50 microliters and the minimal content of the substance estimated was 0.5 microgram/ml. This procedure may be used in screening quick tests of the herbicide concentration in sewage or in extracts from various sources.

[Immunoenzyme analysis of insulin antibodies in human blood serum]. / Immunofermentnyi analiz antitel i insulinu v syvorotke krovi cheloveka.

Indirect solid-phase enzyme immunoassay (EIA) was used for the titration of antibodies to insulin in human blood sera. The modification suggested by the authors permits assessing the immunoglobulins G and M insulin-binding ability. The technique was optimized for all the stages of analysis and a method for estimating the insulin-binding ability of IgG in the sera diluted 1:80 and of IgM in the sera diluted 1:320 was developed. Fifty blood sera of patients with insulin-dependent diabetes mellitus were analyzed. EIA results were in good correlation with the findings of radioimmunoassay.

[Optimization of the conditions for the immunochemical nephelometric analysis of IgG, IgA and IgM]. / Optimizatsiia uslovii immunokhimicheskogo nefelometricheskogo analiza IgG, IgA i IgM.

Immunonephelometric analysis (INA) is widely used in practical chemistry. the authors have examined the possibility of using Soviet reagents in an ICS Analyzer Beckman (USA) nephelometer. The effects of polyethylene glycol concentration, the reaction buffer pH and ionic strength on the results of INA were under study. Optimal conditions for INA with the test systems anti-IgA-IgA, anti-IgG-IgG, and anti-IgM-IgM with Soviet reagents have been developed. Lyophilized immunoreagents from the Mancini radial immunodiffusion kit, prepared at the N. F. Gamaleia Research Institute for Epidemiology and Microbiology, can be employed for analysis with this nephelometer.

[Immunoenzyme method of sequential saturation for determining antigens using the model of insulin]. / Immunofermentnyi metod posledovatel'nogo nasyshcheniia dlia opredeleniia antigenov na modeli insulina.

The method for the determination of insulin by means of the enzyme immunoassay, based on the use of insulin-peroxidase conjugates, has been developed. In this assay the scheme of the successive saturation of the active sites of antibodies is used. The antigenic properties of two conjugates differing in the method of their preparation are compared. The conjugates were obtained by the covalent binding of peroxidase, oxidized in its carbohydrate component, with insulin (conjugate 1) or hexamethylene-diamine-modified insulin (conjugate 2). The conjugates represented a mixture of oligomers differing in their molecular weight. Conjugate 1 possessed higher affinity to antibodies and higher enzymatic activity than conjugate 2. The method for evaluating the quality of antisera to insulin used in the assay has been proposed. The time of the insulin assay is 5-16 hours, the limit of insulin detection is 5 microU/ml, the variation factor is 3-12%.

[Properties of the human anti-IgG-IgG system in various modifications of immunoenzyme analysis]. / Svoistva sistemy anti-IgG-IgG cheloveka v razlichnykh modifikatsiiakh immunofermentnogo analiza.

The physico-chemical characteristics of EIA, and in particular sensitivity, accuracy, reproducibility, can undergo essential changes with variations occurring in the course of the experiment, which makes it necessary to optimize conditions at each stage of the assay. For this purpose the human anti-IgG-IgG system has been studied in two modifications of EIA: the sandwich and competitive methods.

[Use of immunoenzyme analysis for controlling the activity of antisera to insulin]. / Ispol'zovanie immunofermentnogo analiza dlia kontrolia aktivnosti antisyvorotok k insulinu.

Antisera to insulin were obtained from guinea pigs hyperimmunized with the use of Freund's adjuvant. To control the specific activity of antisera, the highly specific modern method of ELISA was used. Insulin was allowed to adsorb on the surface of the wells of polystyrene microplates, then consecutive dilutions of the tested antisera to insulin were placed into the wells. The insulin-antibody complexes thus obtained were detected by means of immunoperoxidase conjugate. Antibody titers in antisera obtained from different animals varied from 5 X 10(-3) to 10(-5). A good correlation between the results obtained by ELISA and by radioimmunoassay was observed.

[The Homogeneous immunocofactor method of determining insulin]. / Gomogennyi immunokofaktornyi metod opredeleniia insulina.

An immunocofactor quantitative method of measuring insulin in solution was developed. The method uses antibody competitive binding of free and NAD labeled insulin. The NAD-insulin conjugate was obtained by covalent binding of the C(6)-aminogroup modified cofactor with insulin by means of soluble carbodiimide. To determine the NAD-insulin conjugate, which was not bound with antibodies, in the presence of antibodies and free insulin, an enzymic system of the cofactor regeneration with conjugated substrates of horse liver alcohol dehydrogenase, cyclohexanol and p-nitroso-N,N-dimethyl aniline, was employed. The sensitivity of insulin assay was about 5.10(-7) M.

[Experimental data on milk casein absorption into the blood in the early postnatal period of development]. / Eksperimental'nye dannye o vsasyvanii kazeinov moloka v krov' v rannii postnatal'nyi period razvitiia.

Blood absorption of unchanged molecules of the food protein casein was studied. Of 18 test serum samples withdrawn from portal blood of a satiated lamb, only 8 contained casein-like protein that had an immunoelectrophoretic mobility of casein and provided a reaction identical to that of casein preparation. Casein hydrolysis products did not interact with anticasein serum. The evidence obtained warrants the conclusion that under normal physiological conditions an insignificant part of casein may be absorbed unchanged by blood from the intestinal lumen during the early postnatal development.

[Amino acid metabolic indices in newborn rats]. / Nekotorye pokazateli aminokislotnogo obmena u novorozhdennykh krys.

In simulated tests on rats of the 1st months of life set up for the purpose of educing causes accounting for a highly effective utilization of the milk proteins in the early post-natal perions subject to study were the amino acids metabolism rates. An investigation of the enzymes activity showed that a falling of the milk proteins utilization effectiveness proceeds against the background of an intensive inclusion of the catabolic route of the amino acids metabolism in the liver. The urea content in the urine correlated with a gradual inclusion of the enzymatic liver systems partaking in the urine formation. The amino acids metabolism with a ramified chain in the liver tissue of rats of the first days of life was found to stand low to a still greater degree than this was the case in adult animals. One of the causes responsible for a highly effective utilization of the milk proteins in the perinatal period may be due to an incomplete functioning of the catabolic route of the amino acids metabolism in the liver.

[Effective utilization of milk proteins by rats during the neonatal period]. / Effektivnost' utilizatsii belkov moloka krysami v period novorozhdennosti.

The assimilation of milk proteins in dynamics by rats during the first 23 days of their life was studied. During this time the animals were kept on a natural milk feeding and were denied the maternal one. Data on the amount of consumed protein and on an increase of the protein content in the carcass in 2- and 4-day long intervals served as a basis for determining the degree of effective utilization of protein (U) by rats. It is shown that the degree of the food protein, while remaining at a high level, experienced certain changes. It is possible to distinguish three periods in the change of this value: the first--starting from birth up to 5 days, with U rising from 74,5 up to 92.1 per cent, the second period, the milk diet proper,--up to the 15th day, marked by a high value of U (approximately 93 per cent) and, finally, the third period--transitional to the ration of an adult animals--marked by a drop of U down to 67.8 per cent. The data obtained may be used in considering issues related to features specific for the processes of the protein assimilationin in the early post-natal period.