[Study of the quality of aminated substrates used for hybridization analysis].

A set of methods for analysis of the quality of aminated substrates that could be a basis for the large-scale manufacturing of biological microchips is suggested. The analysis includes the determination of the number of amino groups, their availability for the immobilization of phosphorylated oligonucleotides, and the characterization of surface properties of the substrates in respect to the nonspecific sorption of reagents during hybridization. A simple procedure was suggested for determination of the density/number of amino groups. It is based on the use of dimethoxytrityl chloride with the subsequent spectrophotometric determination of dimethoxytrityl cation. The availability of amino groups was estimated by covalent attachment of an oligonucleotide probe containing a fluorescently labeled group to the aminated surface and the subsequent comparison of the intensity of fluorescing zones formed on the chip. The sorption properties of the surface were investigated with the help of a model hybridization reaction. A comparative analysis of aminated glasses manufactured by various firms and in our laboratory showed that the glasses with the amino group density from 0.7 to 2.0 groups/nm2 prepared by our procedure have the best properties for the hybridization analysis.

[Calcium induced calcium release from liver mitochondria of spontaneously hypertensive rats].

BACKGROUND: Disturbances of ionic homeostasis of cells and recently discovered cellular energy deficiency due to reduced ATP-synthesizing ability of mitochondria are the most important components of pathogenesis of primary hypertension. Therefore it is essential to elucidate relationship between functioning of ionic transport systems especially that of calcium transport and ATP-synthesizing ability of mitochondria. AIM: To study calcium induced calcium release from liver mitochondria of spontaneously hypertensive rats at various initial calcium concentrations in medium. RESULTS: We observed 30% lowering of maximal level of calcium uptake from the medium with unchanged rate of calcium uptake. Under conditions of calcium overload the rate of calcium release into the medium was 30% higher than in control. INTERPRETATION: These results evidenced for changes of functioning of mitochondrial permeability transition pores (MPT-pores) during calcium overload appearing as acceleration of their switch to high conductance mode and equilibration of all ionic gradients on internal mitochondrial membrane. This can negatively affect ATP-synthesizing ability of mitochondria. Thus the presence of linkage between disturbance of ionic homeostasis and intracellular energy deficit was confirmed.

[Decreased ATP-synthesis ability of brain mitochondria in spontaneously hypertensive rats]. / Snizhennaia ATF-sinteziruiushchaia sposobnost' mitokhondrii kletok golovnogo mozga krys so spontannoi gipertenziei (SHR).

UNLABELLED: It has been shown previously that a decrease of ATP amount and changed balance of other macroergic phosphates occurs in different tissues of spontaneously hypertensive rats (SHR) compared with control normotensive rats (WKY). AIM: To assess the ability of SHR brain isolated mitochondria to synthesize ATP and to elucidate its relation to extramitochondrial calcium concentration. RESULTS: The present work shows for the first time that SHR brain mitochondria initially differ from WKY ones by decreased (by 30%) ATP synthesis rate. When calcium concentration is increased up to 20 or 40 micromole/l ATP synthesis rates in WKY and SHR mitochondria are about one half compared with the condition when the medium does not contain calcium, but the differences between SHR and WKY remain the same. CONCLUSION: Decreased ATP synthesis rate in SHR mitochondria may be considered as the main cause of cell energy deficiency observed in experimental variant of primary arterial hypertension.

[Polymorphism of the central region of D-loop of mitochondrial DNA and personality identification by forensic medicine methods]. / Izuchenie polymorfizma tsentral'nogo regiona D-petli mitokhondrial'noi DNK cheloveka v aspekte sudebno-meditsinskoi identifikatsii lichnosti.

Polymorphism of mDNA D-loop central region (CR), positions 16366-0072) was studied versus hypervariable sections (HVS1), positions 16024-16365, and HVS2, positions 00073-00340, for a sample of 71 residents of the Russian Federation. Ten polymorphic positions with 56 nucleotide substitutions, 55 of which are transitions, were detect in the CR section; no insertions or deletions were found there. It was proven as possible to use the mDNA CR locus as an additional identification marker in the forensic-expert of the mDNA control region. The probability of random coincidence (RC) of haplotypes in joint typing of HVS1, HVS2 and CR made 0.0208, which is 1.4-fold less versus the same parameter for haplotypes HVS1/HVS2 (RC = 0.0284).

[Distribution of HLA DQA1, LDLR, GYPA, HBGG, D7S8, and GC locus alleles in the population of Russia]. / Raspredelenie allelei lokusov HLA DQA1, LDLR, GYPA, HBGG, D7S8, GC sredi naseleniia Rossii.

The distribution of chromosome locus alleles HLA DQA1, LDLR, GYPA, HBGG, D7S8, and GC of PolyMarker molecular genetic individualizing system was studied for the first time in a representative "mean statistical" sampling of Russian population. Typing of these locuses was carried out in 391 donors (no relatives) from 63 regions of the Russian Federation. The incidence of genotypes of all 6 locuses corresponded to the expected values, estimated on the basis of Hardy-Weinberg equilibrium hypothesis. This allows us to use the frequency characteristics of HLA DQA1 locus and the PolyMarker locuses determined in our study as the reference parameters for standard probability estimations in DNA identification. The frequencies of PolyMarker locuses alleles in the Russian sampling (in comparison with other ethnic groups) coincided best of all for allele frequencies in Europeoids living in the USA. For expert evaluation of the efficiency of using these locuses as molecular genetic markers with identification purposes, the discrimination potential was estimated separately for each locus and combinations thereof. HLA DQA1 locus was the most informative of the studied 6 locuses. The main population characteristics of this locus (probability of accidental coincidence, potential of discrimination--PD, polymorphism coefficient--PIC, exclusion potential--Pe, and mean value of parentage index--PI) were estimated for the population of Russia. The frequency distribution of alleles of the studied panel of locuses in the mean statistical Russian population obtained in our study can be used in molecular genetic personality identification and in anthropological studies.

[Localization of disulfide bonds in the alpha 1 acidic glycoprotein molecule and study of their effect on ability of the protein to interact with ethidium bromide]. / Lokalization disul'fidnykh sviazei v molekule kislogo glikoproteina alpha1 i issledovanie ikh vliianiia na sposobnost' belka vzaimodeistvovat' s bromistym étidiem.

alpha 1-Acid glycoprotein (orosomucoid) was purified from the human and murine blood sera using phenol deproteinization. As opposed to the murine protein, the human orosomucoid bound the fluorescent dye ethidium bromide but lost this ability after treatment with beta-mercaptoethanol, which breaks disulfide bonds. Disulfide bonds between the Cys23 and Cys165 residues of the human orosomucoid and between the Cys91 and Cys184 residues of the murine orosomucoid were identified.

[Weight distribution and optical characteristics of soil humic acids fractionated by gel chromatography and electrophoresis]. / Vesovoe raspredeleniie i opticheskiie svoistva pochvennykh guminovykh kislot, fraktsionirovannykh sochetaniem gel'-khromatografii i élektroforeza.

Combination of gel chromatography on Sephadex and polyacrylamide gel electrophoresis was used to obtain molecular weight- and electrophoretic mobility-uniform fractions of humic acids from soil of different types (chernozem, sod-podzol, grey forest, and red soils). Extinction and color (E4/E6) coefficients, as well as weight distribution (%) were studied for all humic acids and their fractions. The fractions differed by the weight distribution (%). Sod-podzol and red soils had twice the amount of high-molecular and half the amount of low molecular HA fractions as compared to chernozem and grey forest soil.

[Thermoinactivation of NAD-kinase from rabbit liver]. / Termoinaktivatsiia NAD-kinazy iz pecheni krolika.

Thermal inactivation of homogeneous NAD-kinase and its partially purified preparation containing NAD-kinase in complex with glutamate dehydrogenase has been studied. The complex is more resistant to thermal inactivation in comparison with isolated NAD-kinase; its inactivation consists in irreversible dissociation preceded by a period of constant activity. No period of constant activity is observed when homogenous NAD-kinase is subjected to thermal inactivation, which represents a two-step process. At the first stage tetramers reversibly dissociate into dimer, while at the second stage the dimers are denatured. The values of activation energy of dissociation and denaturation are equal to 40 and 52 kcal/mol, respectively.

[A new two-stage plan for isolating human alpha1-acidic glycoprotein (orosomucoid). Orosomucoid interacts with ethidium bromide]. / Novaia dvukhstadiinaia skhema vydeleniia chelovecheskogo alpha1- kislogo glikoproteina (orozomukoida). Orozomukoid vzaimodeistvuet s bromistym étidiem.

Human alpha 1-acid glycoprotein (orosomucoid, AGP) was purified to homogeneity by a two-step procedure using phenol and chloroform deproteinization of serum with subsequent preparative electrophoresis on agarose gel. It was obtained 0.15-0.3 mg of protein from 1 ml of serum. Human alpha 1-AGP binds ethidium bromide but not DNA. Neither other human serum proteins nor mice and rat alpha 1-AGP bind ethidium bromide. This property of human orosomucoid depends on its amino acid sequence rather than on the carbohydrate part of the molecule.

[DNA hyperproduction in the blood plasma of animals with experimental diabetes and its compensation using the transplantation of insulin-producing tissue into the anterior chamber]. / Giperproduktsiia DNK v plazme krovi u zhivotnykh s éksperimental'nym diabetom i ee kompensatsiia s pomoshch'iu transplantatsii insulinprodutsiruiushchei tkani v peredniuiu kameru glaza.

Comparative analysis of serum DNA content in healthy rats, rats with experimental alloxan diabetes, and the rats with the diabetes compensated by transplantation of the embryonal pancreas into the anterior chamber of the eye. In the course of diabetes the concentration of free-circulating serum DNA reliably increase 10-12 times, and the compensation of the diabetes returns it to the norm. The revealed relationship may be used for diagnostics of initiation and full compensation of the insulin-dependent diabetes.

[The blood coagulation system under overdoses of the copolymer of 1-vinyl-3-methylimidazole iodide with the diethyl ether of maleic acid demonstrating antiheparin activity]. / Issledovanie sistemy svertyvaniia krovi v usloviiakh peredozirovok sopolimera 1-vinil-3-metilimidazoliia iodida s diétilovym éfirom maleinovoi kisloty, proiavliaiushchego antigeparinovuiu aktivnost'.

The effects of the active heparin antagonist--1-vinyl-3-methylimidazolium iodide-maleic acid diethyl ether copolymer--on the blood coagulative system were investigated. The effect of polycation was shown to appear only when the therapeutical dose was significantly increased and to manifest itself in the development of hypocoagulation. Hypocoagulative changes occurred only by inhibiting the activity of clotting factors V and VIII. Also, there was a moderate reduction in platelet counts.

[The experimental effect of the copolymer of 1-vinyl-3-methyl imidazole iodide with maleic acid diethyl ester on blood coagulability]. / Vliianie sopolimera 1-vinil-3-metil imidazolii iodida s diétilovym éfirom maleinovoi kisloty na svertyvaemost' krovi v éksperimente.

The antiheparin activity and acute toxicity of the copolymer of 1-vinyl-3-methyl inidazol iodide (VMII) with diethyl ether of maleic acid (DEMA) were investigated. It was shown that the injection of DEMA group to the molecule of VMII polymer was adequate for storage of a high antiheparin activity of the polymer with the structure. This composition of the copolymer has a low acute toxicity and high selectivity to heparin in the blood. Thrombocytopenia effect is absent.

[The nucleoprotein fraction of mouse serum in normal and pathological processes]. / Issledovanie nukleoproteinovoi fraktsii syvorotki krovi myshei v norme i pri patologicheskikh protsessakh.

Analysis of the blood serum of healthy and infected with malaria plasmodium mice showed a steep rise in content of linear double-stranded DNA (0.2-0.5 and 2-15 gamma/ml, respectively). Some physico-chemical properties of serum DNA and a DNA-associated glycoprotein (M approximately 40 kDa) are determined.

[Analysis of the trypsin hydrolysate of a 40-kDa protein, found in the nucleoprotein of serum. Identification of it as alpha(1)-acidic glycoprotein (orosomucoid)]. / Analiz produktov tripticheskogo gidrolizata 40-kDa belka, vkhodiashchego v sostav nukleoproteinovoi fraktsii syvorotki krovi. Identifikatsiia ego v kachestve alpha(1)-kislogo glikoproteina (orozomukoida).

From the tryptic hydrolysis of 40-kDa protein from the mouse blood serum thirty-four peptides were isolated by HPLC, of which complete and partial amino acid sequence was established for twenty-eight and six, respectively. On the basis of these data the protein is identified as the blood serum alpha 1-acid glycoprotein.