[Assessment of relationship of lipid and carbon profile indicators with vitamin D supply in children depending on body mass index].

Overweight children represent a particularly vulnerable group for hypovitaminosis D. Clinical studies on the relationship between vitamin D (VD) deficiency and metabolic risk factors for cardiovascular disorders are controversial, and for children of primary school age who have overweight and obesity are insufficient. The aim of the research was to study the relationship between lipid and carbohydrate metabolism indicators and VD status in children, depending on the body mass index. Material and methods. A cross-sectional (one-step) study was carried out on a sample of 154 children with different weight of 8-10 years old (74 girls, 80 boys). Three groups of research participants were identified: group 1 - 44 obese, group 2 - 58 overweight, group 3 - 52 children with normal body weight. For all children, the serum level of 25(OH)D, parathyroid hormone (PTH), calcium (Ca), phosphorus (P), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG), ß-lipoproteins, glucose, insulin was determined, and Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) was also calculated. Results. VD deficiency in obese children was found almost 2.3 fold more often than in overweight (p=0.002) and 2.8 fold more often than in children with normal body weight (p=0.001). Indicators of lipid and carbohydrate metabolism were within physiological limits. However, in obese children they significantly exceeded the indicator of healthy children (p<0.05). When comparing the results of biochemical studies, it was revealed that children with VD deficiency [25(OH)D <20 ng/ml] had statistically significantly higher medians of PTH, TC, TG, ALT, AST, glucose, insulin, HOMA-IR and lower P and Ca level compared with children with normal micronutrient blood content (p<0.05). The medians of ALT, AST, TC, ß-lipoproteins, TG, glucose, insulin and HOMA-IR levels in obese children with VD deficiency were statistically significantly higher than in children with normal body weight and VD deficiency and in healthy children with an optimal concentration of 25(OH)D. At the same time, there was no statistically significant difference between the indicators of lipid and carbohydrate metabolism in the group of healthy children with normal VD status and its deficiency. Conclusion. VD deficiency is an important predictor of obesity complications and it exacerbates the risk of cardiometabolic disorders in children who are obese in the early school years.

[Markers of the course of obliterating atherosclerosis of lower limb arteries]. / Markery techeniia obliteriruiushchego ateroskleroza arterii nizhnikh konechnostei.

AIM: The purpose of the study was to determine the level of substances damaging the vascular endothelium, as well as to assess their effect on the functional state of the endothelium and the course of obliterating atherosclerosis of lower limb arteries. PATIENTS AND METHODS: The study included a total of 112 people, subdivided into three groups: those with an unfavourable course of obliterating atherosclerosis of lower limbs arteries (n=48) - group 1, patients with obliterating atherosclerosis of lower limbs arteries with a conventionally favourable course (n=48) - group 2, and apparently healthy volunteers (n=16). In all subjects, the following parameters were analysed: stable metabolites of nitric oxide II, endothelin-1, homocysteine and basal insulin. RESULTS: The level of stable nitric oxide metabolites (p<0.001 as compared with group 1; p<0.045 compared with group 2) was lower in the groups of patients with obliterating atherosclerosis of lower limb arteries (88.5±7.3 µmol/L in group 1; 161.5±8.6 µmol/L in group 2) as compared with healthy volunteers (226.0±28.6 µmol/L). In its turn, the level of nitric oxide was statistically significantly lower (p<0.001) in group 1 patients as compared with those of group 2. The level of endothelin-1 turned out to be higher (p<0.001) in group 1 (2.1±0.1 ng/ml) as compared with group 2 (1.6±0.1 ng/ml). Comparing group 1 patients with healthy volunteers (1.4±0.1 ng/ml), the level of endothelin-1 had also higher values (p<0.001). The level of endothelin-1 did not differ (p=0.270) as compared with group 2 and healthy volunteers. Comparing the homocysteine level in patients of the examined groups (20.7±0.8 µmol/L in group 1 patients and 18.1±0.6 µmol/L in group 2 patients) with healthy volunteers (13.0±0.4 µmol/L) demonstrated an increase in the parameters (p<0.001). The level of homocysteine turned out to be higher in group 1 patients than in those of group 2 (p<0.001). The level of basal insulin turned out to be significantly higher in the studied groups of patients with obliterating atherosclerosis of lower limb arteries (24.9±4.6 mIU/L in group 1; 8.0±0.7 mIU/L in group 2) than in healthy volunteers (5.1±0.5 mIU/L). Statistically significant (p<0.001) hyperinsulinemia was observed in group 1 as compared with group 2. CONCLUSION: Hyperhomocysteinemia and hyperinsulinemia are predictors of an unfavourable course of the disease. In a high level of these parameters, one may predict an unfavourable course of obliterating atherosclerosis of lower limb arteries.

Role of NUDIX Hydrolases in NAD and ADP-Ribose Metabolism in Mammals.

Proteins of the NUDIX hydrolase (NUDT) superfamily that cleave organic pyrophosphates are found in all classes of organisms, from archaea and bacteria to higher eukaryotes. In mammals, NUDTs exhibit a wide range of functions and are characterized by different substrate specificity and intracellular localization. They control the concentration of various metabolites in the cell, including key regulatory molecules such as nicotinamide adenine dinucleotide (NAD), ADP-ribose, and their derivatives. In this review, we discuss the role of NUDT proteins in the metabolism of NAD and ADP-ribose in human and animal cells.

Crystal structures of two dimeric nickel di-phenyl-acetate com-plexes.

In the crystal structures of the title com-pounds, namely µ-aqua-κ2 O:O-di-µ-di-phenyl-acetato-κ4 O:O'-bis-[(di-phenyl-acetato-κO)bis-(pyridine-κN)nickel(II)], [Ni2(C14H11O2)4(C5H5N)4(H2O)] (1) and µ-aqua-κ2 O:O-di-µ-di-phenyl-acetato-κ4 O:O'-bis-[(2,2'-bi-pyridine-κ2 N,N')(di-phenyl-acetato-κO)nickel(II)]-aceto-nitrile-di-phenyl-acetic acid (1/2.5/1), [Ni2(C14H11O2)4(C10H8N2)2(H2O)]·2.5CH3CN·C14H12O2 (2), the com-plex units are stabilized by a variety of intra- and inter-molecular hydrogen bonds, as well as C-H⋯π and π-π contacts between the aromatic systems of the pyridine, dipyridyl and di-phenyl-acetate ligands. Despite the fact that the di-phenyl-acetate ligand is sterically bulky, this does not inter-fere with the formation of the described aqua-bridged dimeric core, even with a 2,2'-bi-pyridine ligand, which has a strong chelating effect.

The Regulatory Role of NAD in Human and Animal Cells.

Nicotinamide adenine dinucleotide (NAD) and its phosphorylated form NADP are the major coenzymes in the redox reactions of various essential metabolic pathways. NAD+ also serves as a substrate for several families of regulatory proteins, such as protein deacetylases (sirtuins), ADP-ribosyltransferases, and poly(ADP-ribose) polymerases, that control vital cell processes including gene expression, DNA repair, apoptosis, mitochondrial biogenesis, unfolded protein response, and many others. NAD+ is also a precursor for calcium-mobilizing secondary messengers. Proper regulation of these NAD-dependent metabolic and signaling pathways depends on how efficiently cells can maintain their NAD levels. Generally, mammalian cells regulate their NAD supply through biosynthesis from the precursors delivered with the diet: nicotinamide and nicotinic acid (vitamin B3), as well as nicotinamide riboside and nicotinic acid riboside. Administration of NAD precursors has been demonstrated to restore NAD levels in tissues (i.e., to produce beneficial therapeutic effects) in preclinical models of various diseases, such as neurodegenerative disorders, obesity, diabetes, and metabolic syndrome.

[Sex differences of P-glycoprotein functional activity and expression in rabbits].

The research consists in the investigation of the sex differences of P-glycoprotein functional activity and expression in Chinchilla rabbits. P-glycoprotein functional activity was assessed by the pharmacokinetics of its probe substrate--fexofenadine after its single oral administration. P-glycoprotein expression was investigated by immunohistochemistry method. It is shown that male's maximal concentration of fexofenadine, its areas under concentration-time curves, half-life and retention time were higher and its clearance was lower than female's. The efficient differences in pharmacokinetic parameters of fexofenadine confirm more intensive excretion and less intensive absorption in gastro-intestinal tract of fexofenadine. This data indicate that P-glycoprotein activity is more active in female than in male. Immunohistochemistry analysis shows that total liver and intestine P-glycoprotein expression is more intensive in females, than in males that correlates with its more active functioning.

[Hepatoprotectors in treatment of alcoholic liver disease].

The best hepatoprotective effect has combined appointment Essentiale forte N, Heptral and Apilak at rats with chronic alcohol intoxication. Use of these drugs is characterized by minor morphological changes in structure of liver and constant biochemical parameters.

[Studies in chicken genetics: commemorating the 120th anniversary of the outstanding Soviet geneticist A. S. Serebrovsky (1892-1948)].

The paper highlights the research of A. S. Serebrovsky in chicken genetics, including gene mapping and inheritance of morphological traits. Genetic formulas for several breeds are presented. The data of genetic surveys for local chicken populations from 23 regions of the former Soviet Union are also reviewed. The personal data of the authors on the morphotypological characteristics of different chicken breeds are given and discussed.

[Mitochondrial and lysosomal pathways of death of hepatocytes of lamprey Lampetra fluviatilis L].

Mechanisms of mitochondrial and lysosomal pathways of natural cell death in lamprey hepatocytes at the spring period of prespawning migration are described. The mitochondrial pathways (release of cytochrome c from mitochondria into cytosol and activation ofcaspases) operates according to the classical scheme known for apoptosis. The lysosomal cell death pathway connected with activation of cathepsin B has been revealed quite recently in cells in pathologies, in particular at obstruction of gallbladder and bile ducts. The peculiarity of lamprey hepatocytes consists in biliary atresia (the absence both of gallbladder and of bile ducts) in liver of adult animals. Thereby the lamprey hepatocytes represent an excellent object for study of this new pathway of cell death. We have revealed a parallel development of the mitochondrial and lysosomal pathways of cell death of lamprey hepatocytes.

[Accumulation of sodium and potassium ions in oocytes of the river lamprey Lampetra fluviatilis during prespawning period].

Accumulation of Na+ and K+ ions in oocytes of the river lamprey Lampetra fluviatilis and their transport across the plasma membrane is realized by two main mechanisms--Na,K-pump and Na,K,Cl-cotransport. At the prespawning period from December to May the intracellular Na+ concentration was observed to increase from 10 to 25 mM and the K+ concentration--from 28 to 45 mM. Results obtained on isolated oocytes with aid of 204Tl radioactive label have shown that contributions of the Na,K-pump and Na,K,Cl-cotransport to potassium accumulations were close until March. In spring, the total K+ inflow almost doubled owing to activation of the Na,K-pump, whereas contribution of Na,K,Cl-cotransport did not change. It seems that an increase of the intracellular content of the main inorganic cations in oocytes resulted in parallel activation of the Na,K-pump and probably of Na/H-exchange. The biological significance of activation of these mechanisms of ion transport at the prespawning period might be due to a necessity of accumulation of Na+ and K+ ions in concentrations optimal for subsequent embryonic development.

[Transport of monovalent thallium across the membrane of oocyte of the lamprey Lampetra fluviatilis].

Mechanisms of transport of monovalent thallium across the membrane of oocyte of the lamprey Lampetra fluviatilis were studied by using 204Tl. Transport of Tl+ in lamprey oocytes has been shown to be realized by at least two pathways: through Na/K-pump and by the mechanism of Na,K,Cl-cotransport. In the standard Ringer solution (mM): 4 KCl, 140 NaCl, 0.5 CaCl2, 5 glucose, 10 Tris-HCl--in the presence of oubain, the coefficient of the 204Tl stationary distribution (cell/medium) was within the range of 2.3-2.5, while the time necessary to reach its 50 % value amounted to 40-5 min at 20 degrees C. In potassium-free media, transport of 204Tl via Na/K-pump was described by simple kinetis with saturation and was characterized by the value V(max) = 520 pmol/(cell x h) and K(M) = 0.3 mM. In the presence of 4 mM K+ and 0.1 mM/l Tl+, the oubain-sensitive Tl+ flow decreased to 75 pmol/(cell x h). At activation of the mechanism of Na,K,Cl-cotransport by the outer Na+ (in Na-NMDG media of different composition) the total inflow of Tl+ reached 193 +/- 20 pmol/(cell x h), while the butamenide-sensitive component--119 +/- 12 pmol/(cell x h) with K(M) for Na+ about 20 mM. In the incubation media with variable concentration of chloride ions (replacement of Cl- by NO3(-)) the total Tl+ flow reached 220 +/- 21, while via the mechanisms of Na,K,Cl-cotransport--87 +/- 8 pmol/(cell x h). Under our experimental conditions, mechanisms of active transport and Na,K,Cl-cotransport accounted for 94% of the Tl+ inflow. The potassium channels that usually are also permeable to monovalent thallium ions were not revealed.

[Dynamics of some postreplication DNA repair proteins in carcinogen-damaged mammalian cells]. / Issledovanie dinamiki nekotorykh belkov postreplikativnoi reparatsii DNK v kletkakh mlekopitaiushchikh posle obrabotki kantserogenami.

Many types of DNA lesions in template strands block DNA replication and lead to a stalling of replication forks. This block can be overcome (bypassed) by special DNA polymerases (for example, DNA polymerase eta, Pol eta) that perform translesion synthesis on damaged template DNA. The phenomenon of completing DNA replication, while DNA lesions remain in the template strands, has been named post-replication repair (PRR). In yeast Saccharomyces cerevisiae, PRR includes mutagenic and error-free pathways under the regulation of the RAD6/RAD18 complex, which induces ubiquitylation of PCNA. In mammalian cells, Pol eta accumulates in replication foci but the mechanism of this accumulation is not known. Pol eta possesses a conserved PCNA binding motif at the C terminal and phosphorylation of this motif might be essential for its interaction with PCNA. We have shown previously that staurosporine, an inhibitor of protein kinases, inhibits PRR in human cells. In this study we examined whether the accumulation of Pol eta in replication foci after DNA damage is dependent on phosphorylation of the PCNA binding motif. We also studied DNA damage-induced phosphorylation of GFP-tagged human Rad18 (hRad18) and its accumulation in replication foci. Our data indicate that (1) Pol eta is not phosphorylated in response to UV irradiation or MMS treatment, but its diffusional mobility is slightly decreased, and (2) hRad18 accumulates in MMS-treated cells, and considerable amount of the protein co-localizes with detergent insoluble PCNA in replication foci; these responses are sensitive to staurosporine. Our data suggest that hRad18 phosphorylation is the staurosporine-sensitive PRR step.

Dephosphorylation of histone gamma-H2AX during repair of DNA double-strand breaks in mammalian cells and its inhibition by calyculin A.

The induction of DNA double-strand breaks (DSBs) by ionizing radiation in mammalian chromosomes leads to the phosphorylation of Ser-139 in the replacement histone H2AX, but the molecular mechanism(s) of the elimination of phosphorylated H2AX (called gamma-H2AX) from chromatin in the course of DSB repair remains unknown. We showed earlier that gamma-H2AX cannot be replaced by exchange with free H2AX, suggesting the direct dephosphorylation of H2AX in chromatin by a protein phosphatase. Here we studied the dynamics of dephosphorylation of gamma-H2AX in vivo and found that more than 50% was dephosphorylated in 3 h, but a significant amount of gamma-H2AX could be detected even 6 h after the induction of DSBs. At this time, a significant fraction of the gamma-H2AX nuclear foci co-localized with the foci of RAD50 protein that did not co-localize with replication sites. However, gamma-H2AX could be detected in some cells treated with methyl methanesulfonate which accumulated RAD18 protein at stalled replication sites. We also found that calyculin A inhibited early elimination of gamma-H2AX and DSB rejoining in vivo and that protein phosphatase 1 was able to remove phosphate groups from gamma-H2AX-containing chromatin in vitro. Our results confirm the tight association between DSBs and gamma-H2AX and the coupling of its in situ dephosphorylation to DSB repair.

[Genetic polymorphism of Russian, European, and Asian chicken breeds as revealed with DNA and protein markers]. / Geneticheskii polimorfizm russkikh, evropeiskikh i aziatskikh porod kur, vyiavliaemyi s pomoshch'iu DNK i belkovykh markerov.

The variation in polymorphic DNA (RAPD and minisatellite) and protein markers was compared for nine Russian chicken breeds differing in morphological and productivity types and in origin, three European egg breeds, and three broiler breeds of the Asian origin. Genetic diversity indices were calculated for each breed group and each marker type and were used to construct dendrograms of genetic similarity. In all breed groups, minisatellites and RAPD markers revealed higher genetic diversity as compared with protein markers. With any type of markers, genetic diversity of the Russian and Asian broiler breeds proved to be significantly higher than that of the European egg breeds. The differentiating potentialities of molecular and genetic biochemical markers at the breed level and the origin of the Russian chicken breeds are discussed.

[UV-induced immobilization of DNA repair protein XPA in different human cell line]. / UF-indutsirovannaia immobilizatsiia reparatsionnogo belka XPA v raz lichnykh liniiakh kletok cheloveka.

XPA repair protein is absolutely needed for nucleotide excision repair (NER). It preferentially binds UV-irradiated DNA in vitro and possibly takes place in the recognition of pyrimidine dimers, the main type of UV-lesions in DNA. Using immunofluorescent microscopy and immunoblotting technique we have found that XPA protein is fully extractable by Triton X-100 solution from non-irradiated normal human fibroblasts, but after UV-irradiation its extractability decreases in UV-dose dependent manner. UV-induced XPA-immobilization was observed in human cell lines with different types of repair defects, but XPA-extractability from unirradiated cells of these lines was significantly lower in comparison with normal fibroblasts. These data do not permit to make conclusion concerning the distinct connection of this phenomenon with different pathways of NER. Histone deacetylase inhibitor, sodium butyrate, did not change the level of extractability in unirradiated and UV-irradiated normal human cells and CHO cells, defective in global genome repair, that indicated the independence of XPA-immobilization from the level of histone acetylation. It was established with the help of confocal microscopy that XPA-foci in detergent-treated UV-irradiated cell were partially colocalized with the focal sites of PCNA, an auxiliary protein of DNA polymerases delta and epsilon. It may mean that a part of detergent-resistant XPA foci correspond to DNA repair synthesis sites, but the major part of immobilized XPA reflects the early step of repair proteins assembly formation needed for the repair of the lesions.