[Organization of maxillofacial and dental care for the wounded of the Voronezh front (July 1942 - July 1943)]. / Organizatsiya chelyustno-litsevoi i stomatologicheskoi pomoshchi ranenym Voronezhskogo fronta (iyul' 1942 g. ­ iyul' 1943 g.).

The article is devoted to the history of the formation of the system of assistance to maxillofacial wounded soldiers of the Voronezh Front during the battles for Voronezh in 1942-1943. The difficulties and achievements in the implementation of phased assistance to the wounded in the face, as well as the improvement of the organizational structure of surgical and dental care for soldiers and officers of the front are reflected.

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MicroRNAs epigenetically regulate physiological and pathological processes. Previously, we found that miR-204-5p is expressed at low levels in melanoma cells, and an increase in its level leads to a change in proliferation, migration, and invasion of these cancer cells. Now, using bioinformatics analysis, it has been shown that the target of miR-204-5p is FOXC1 transcription factor, which is implicated in carcinogenesis. Using the luciferase reporter assay, it was found that miR-204-5p suppresses expression of the FOXC1 gene by binding to its 3' non-coding region. Transfection of small interfering RNA (siRNA) targeting FOXC1 into melanoma cells caused a decrease in miR-204-5p levels, which is consistent with the generally accepted concept of feedback regulation of miRNA expression by target genes. According to the results of the MTT test and fluorescence microscopy, the proliferation level of melanoma cells under the influence of siRNA to FOXC1 decreased 72 h after transfection. Changes in the ratio of cells by cell cycle phase were analyzed using flow cytometry. Regulatory relationships between FOXC1 and miR-204-5p, and an inhibitory effect of FOXC1 knockdown on melanoma cell proliferation were revealed. Based on the results, it can be assumed that miR-204-5p regulates proliferation of melanoma cells by affecting FOXC1 expression.

Coelenterazine-dependent luciferases.

Bioluminescence is a widespread natural phenomenon. Luminous organisms are found among bacteria, fungi, protozoa, coelenterates, worms, molluscs, insects, and fish. Studies on bioluminescent systems of various organisms have revealed an interesting feature - the mechanisms underlying visible light emission are considerably different in representatives of different taxa despite the same final result of this biochemical process. Among the several substrates of bioluminescent reactions identified in marine luminous organisms, the most commonly used are imidazopyrazinone derivatives such as coelenterazine and Cypridina luciferin. Although the substrate used is the same, bioluminescent proteins that catalyze light emitting reactions in taxonomically remote luminous organisms do not show similarity either in amino acid sequences or in spatial structures. In this review, we consider luciferases of various luminous organisms that use coelenterazine or Cypridina luciferin as a substrate, as well as modifications of these proteins that improve their physicochemical and bioluminescent properties and therefore their applicability in bioluminescence imaging in vivo.

THE ANALYSIS OF MAJOR RISK FACTORS OF CARDIOVASCULAR DISEASES AMONG INHABITANTS OF THE NORTH REGION.

Last years the problem of organism's adaptation to severe climate-environmental conditions of the Far North has been intensively developed. The Republic of Sakha (Yakutia) is the most northern republic of the Russian Federation. People have created a unique way of life, language, original culture on this cold part of the earth and have carried centuries later. This unique experience has been saved up throughout many centuries and generated in natural environment of habitation and passed from generation to generation. Last years the changes of living conditions of indigenous population, urbanization and globalisation, deterioration of ecological conditions exhausted reserve possibilities of organism. Among the indigenous population health change has menacing character, especially among the children's population. The analysis of major risk factors of the development of cardiovascular diseases among the indigenous population of the north has been carried out in this research.

Ca(2+)-triggered coelenterazine-binding protein from Renilla as an enzyme-dependent label for binding assay.

The recombinant Ca(2+)-triggered coelenterazine-binding protein (CBP) from Renilla muelleri was investigated as a biospecifically labeled molecule for in vitro assay applications. The protein was shown to be stable in solutions in the frozen state, as well as stable under heating and to chemical modifications. Conjugates with biotin, oligonucleotide, and proteins were obtained and applied as biospecific molecules in a solid-phase microassay. CBP detection was performed with intact (no modifications were made) Renilla luciferase in the presence of calcium, and the detection limit was found to be 75 amol. Model experiments indicate that this approach shows much promise, especially with regard to the development of multianalytical systems.

[Calcium-regulated photoproteins of marine coelenterates].

Ca(2+)-regulated photoproteins are bioluminescent proteins responsible for bioluminescence of marine coelenterates. The photoprotein molecule is a stable enzyme-substrate complex consisting of a single polypeptide chain and an oxygen "pre-activated" substrate, 2-hydroperoxycoelenterazine, which is tightly but non-covalently bound with a protein. The bioluminescence is triggered by calcium ions and originates from an oxidative decarboxylation of a protein bound substrate. The review provides current data on the photoproteins structure, the mechanism of bioluminescent reaction, the function of some amino acid residues of an active site in the catalysis and the formation of the emitter, as well as on applications of these proteins in a bioluminescent analysis.

Structural basis for the emission of violet bioluminescence from a W92F obelin mutant.

Mutation of the Trp92 that is known to lie within the active site of the photoprotein obelin from Obelia longissima, results in a shift of the bioluminescence color from blue (lambda(max)=485 nm) to violet. The corrected spectrum shows a new band with lambda(max)=410 nm now contributing equally to the one at longer wavelength. The crystal structure of this W92F obelin determined at 1.72 A resolution shows that there is no significant change in the dimensions of the active site between WT obelin (recombinant Ca2+-regulated photoprotein from Obelia longissima) and the mutant. It is proposed that the bioluminescence spectral shift results from removal of a hydrogen bond from the indole of W92 nearby a hydroxyl belonging to the 6-phenyl substituent of the substrate coelenterazine. Propagation of this change through a conjugated bond system in the excited state of the product coelenteramide affects the coupling of the N1-position and the hydrogen-bonded Y138.

Obelin mRNA--a new tool for studies of translation in cell-free systems.

Obelin mRNA obtained in vitro with the aid of SP6 RNA polymerase was translated in a wheat germ cell-free system. Only the polypeptide with a molecular mass of about 20 kDa was synthesized. The activation of apoobelin with a synthetic coelenterazine revealed a luminescence activity initiated by calcium. The specific activity was 3.6 +/- 0.4 x 10(15)photons per mg of the in vitro synthesized obelin (k=6.9s(-1)). The luminescence of the obelin was in a good correlation with the protein concentration calculated by the incorporation of [14C]Leu. The determination of the amount of de novo synthesized obelin based on measurement of its luminescence is one-thousand times more sensitive than the approach based on the incorporation of labeled amino acid. Thus, obelin mRNA has some advantages for evaluating the efficiency of cell-free translation when compared with standard methods.

Mn(2+)-activated luminescence of the photoprotein obelin.

The light emission of obelin may be initiated by Mn2+ under alkaline conditions. The luminescence takes place in a pH range from 7 to 12 with a sharp optimum at 11.75. The first-order rate constant for Mn(2+)-activated luminescence decay is more than 9 s-1, while that for Ca(2+)-activated luminescence decay is only 6.9 s-1. The Mn2+ concentration-effect curve for obelin determined with simple dilutions of manganese salt is a sigmoid curve. The slope of the curve is moderately dependent on the pH and was not more than 1 within the pH range tested. The maximal light emission, which is initiated by 3.6 x 10(-5) M Mn2+ at pH 11.75 was about 10% of the maximal Ca(2+)-activated luminescence. Mg2+ ions inhibit the Mn(2+)-activated luminescence of obelin. The addition of OH. and O2- scavengers did not influence the Mn(2+)-activated luminescence, but when singlet oxygen quenchers were added, the Mn(2+)-dependent light emission was inhibited. This suggests that the 1O2 might be formed and itself be responsible for chromophore oxidation attended with light emission. NEM and Na2S2O4 inhibit the Mn(2+)-initiated light emission of obelin completely, showing that endogenous hydroperoxide and SH-group(s) of the photoprotein are essential for both Ca(2+)-activated and Mn(2+)-activated light emission of obelin.

[The hormonal function of the fetoplacental system in pregnant women with chronic alcoholic intoxication]. / Gormonal'naia funktsiia feto-platsentarnoi sistemy u beremennykh s khronicheskoi alkogol'noi intoksikatsiei.

Radioimmunoassay was used for the assessment of serum hormone levels. Dysfunctions in the fetoplacental system (decreased levels of progesterone and estrogens) were revealed as were elevated levels of somatotropic hormone, which were in inverse proportion to the fetal weight, in 20 pregnant females who abused alcohol during the entire pregnancy. It was suggested that higher production of somatotropic hormone could occur in the presence of hypophyseal tension which had been evidently developed before the pregnancy.

Insertion of short hepatitis virus A amino acid sequences into poliovirus antigenic determinants results in viable progeny.

In an infectious poliovirus cDNA construct, the determinant encoding antigenic epitope N-Ag1 (in a loop located between two beta-strands in poly-peptide VP1) was altered by site-directed mutagenesis, to be partially similar with the determinants for presumptive epitopes in polypeptides VP1 or VP3 of hepatitis A virus (HAV). The modified constructs proved to be infectious. However, another construct, in which the same locus encoded a 'nonsense' and a relatively hydrophobic amino acid sequence, exhibited no infectivity. These data showed the feasibility of the insertion of foreign sequences in a specific antigenically active locus of the poliovirus icosahedron, and suggest some limitations with respect to the sequences to be 'transplanted'.

[A rapid radioenzyme method for determining dopamine and 3,4-dihydroxyphenylacetic acid in the basal ganglia of the rat brain]. / Bystryi radiofermentnyi metod opredeleniia dofamina i 3,4-dioksifeniluksusnoi kisloty v bazal'nykh gangliiakh mozga krys.

A rapid and high sensitive radioenzymatic method of determination in the rat brain basal ganglia of dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DHPA) whose methylated derivatives were divided by extraction with organic solvents is proposed. The method sensitivity for DA is 0.25 ng in a sample and for DHPA 0.1 ng in a sample. The yield of the internal standard is 80-85%. The method is high specific, the cross reaction during determination of DA and DHPA does not exceed 5%. The study of the effects of some dopaminergic agents (apomorphine, haloperidol. L-DOPA and pargyline on the contents of DA and DHPA in the rat brain basal ganglia in vivo confirms a high specificity of the method. The method may be used for screening of pharmacological compounds possessing the suggested dopaminergic and antimonoamine oxidase activity.

[Reduction of the functional activity of the calcium pump of the sarcoplasmic reticulum of skeletal muscles in response to cold]. / Snizhenie funktsional'noi aktivnosti kal'tsievogo nasosa sarkoplazmaticheskogo retikuluma skeletnykh myshts pod vliianiem kholoda.

Fragments of sarcoplasmic reticulum (SR) from the white skeletal muscles of rat in cooling (1.5 day; -9 to +2 degrees C) revealed a decreased activity of the Ca2+-pump and rate of Ca2+ accumulation. The SR from rats adapted to cold for 2 and 4 weeks, revealed the same decreased functional ability. The Ca/ATP ratio was unchanged in all experiments.