Polymorphic variants of glutamate receptor (<Emphasis Type="Italic">GRIK5</Emphasis>, <Emphasis Type="Italic">GRIN2B</Emphasis>) and serotonin receptor (<Emphasis Type="Italic">HTR2A</Emphasis>) genes are associated with chronic

Chronic obstructive pulmonary disease (COPD) is a complex chronic inflammatory disease of the respiratory system that affects primarily distal respiratory pathways and lung parenchyma. Smoking tobacco is a major risk factor for COPD. The relationship of HTR4 (rs3995090), HTR2A (rs6313), GRIK5 (rs8099939), GRIN2B (rs2268132), and CHRNB4 (rs1948) gene polymorphisms and COPD, as well as the contribution of these polymorphisms to the variations in quantitative characteristics that describe respiratory function, smoking behavior, and nicotine dependence was assessed in an ethnically homogeneous Tatar population. The polymorphisms of HTR2A (rs6313) (P = 0.026, OR = 1.42 for the CC genotype) and GRIN2B (rs2268132) (P = 0.0001, OR = 2.39 for the TT genotype) were significantly associated with increased risk of COPD. The AA genotype of GRIK5 (rs8099939) had a protective effect (P = 0.02, OR = 0.61). Importantly, the HTR2A (rs6313), GRIN2B (rs2268132), and GRIK5 (rs8099939) polymorphisms were only associated with COPD in smokers. Smoking index (pack-years) was significantly higher in carriers of the GRIK5 genotype AC (rs8099939) (P = 0.0027). The TT genotype of GRIN2B (rs2268132) was associated with COPD in subjects with high nicotine dependence according to the Fagerström test (P = 0.002, OR = 2.98). The TT genotype of HTR2A (rs6313) was associated with a reduced risk of the disease in the group with moderate nicotine dependence (P = 0.02, OR = 0.22). The CC genotype of HTR2A (rs6313) and the TT genotype of GRIN2B (rs2268132) were associated with higher levels of nicotine dependence according to the Fagerström test (P = 0.0011 and P = 0.037). Our results may provide insight into potential molecular mechanisms that involve the glutamate (GRIK5, GRIN2B) and serotonin (HTR2A) receptor genes in the pathogenesis of COPD.     

Genotypes of the vitamin D-binding protein gene in patients with chronic obstructive pulmonary disease and in the healthy population of the Republic Bashkortostan

Allele and genotype frequency distributions of the vitamin D-binding protein gene (DBP) were studied in patients with chronic obstructive pulmonary disease (COPD, N = 298) and healthy individuals (N = 237) from two ethnic groups (Tatars and Russians) resident in the Republic Bashkortostan. The DBP genotype frequency distribution significantly differed between Tatars and Russians (X ² = 8.854, df = 5, P = 0.04). The DBP allele frequency distribution was similar in healthy subjects of both ethnic groups, with allele frequency decreasing as GC*1S > GC*1F > GC*2. The most common DBP genotype was GC*1F/1S in Tatars (36.79%) and GC*1S/2 in Russians (34.62%). It was demonstrated that, in Tatars, the genotype GC*1F/1S is protective against COPD, its frequency being significantly lower in COPD patients than in healthy subjects (19.85% vs. 36.79%; X ² = 7.622, P = 0.0067, P _cor = 0.0335; OR = 0.42, 95%CI 0.42–0.95). On the other hand, the genotype GC*1F/2 was more common among COPD patients than among healthy individuals (19.08% vs. 8.49%; X ² = 4.52, P = 0.033, P _cor = 0.165; OR = 2.54, 95%CI 1.067–6.20). No differences in DBP genotype and allele frequency distributions was found between COPD patients and healthy individuals in the Russian population.     

Quantitative estimation of gibberellic acid by paper chromatography

Метод для количественного определения от gibberellic кислоты в процессе брожения средства массовой информации, с использованием бумаги по убыванию хроматографии в butylacetate воды описана. Образца корректируется, чтобы рН 2.5-3.0, добыто с н-бутанола, и 0,05 мл. органического слоя пятнами на Хроматографический бумагу. После equilibration от Атмосфера в банке, chromatogram Разработана в butylacetate насыщенных с водой, за 7 часов, и растворитель разрешено покинуть капельного нижней части листа. Обнаружение осуществляется путем опрыскивания с 3% раствор серной кислоты в метаноле и после сушки бумаги, пятна с синий u.v. флуоресценции наблюдается. Определенный артикль площадь пятна оценивается с помощью калибровочной кривой, заговор с ценностей, стандартов, соответствующих 20, 60 и 120 μ g. gibberellic кислоты. Погрешность оценки составляет ± 10-15% когда оценки выполняются тщательно. Низкий предел чувствительности 5 μg.     

Reasons and limits of substrate activity of modified L-dNTP in DNA biosynthesis.

Theoretical and experimental analysis of interaction of modified D- and L- dNTP as substrates for template-dependent and template-independent DNA polymerases was performed. It is shown that if the modified nucleoside 5'-triphosphates do not contain a substituent in position 3' DNA chains can be extended by both strereoisomeric series with the same kinetic parameters. But the presence of even a 3'-hydroxy group in L-dNTP prevents their incorporation into the DNA chain.     

Proteolytic cleavage of pertussis toxin S1 subunit is not essential for its activity in mammalian cells

Background  

Pertussis toxin (PT) is an exotoxin virulence factor produced by Bordetella pertussis, the causative agent of whooping cough. PT consists of an active subunit (S1) that ADP-ribosylates the alpha subunit of several mammalian G proteins, and a B oligomer (S2–S5) that binds glycoconjugate receptors on cells. PT appears to enter cells by endocytosis, and retrograde transport through the Golgi apparatus may be important for its cytotoxicity. A previous study demonstrated that proteolytic processing of S1 occurs after PT enters mammalian cells. We sought to determine whether this proteolytic processing of S1 is necessary for PT cytotoxicity.     

DNA repair XPCC1 and XPD genes polymorphism as associated with the development of bladder cancer and renal cell carcinoma

We examined the associations between the polymorphic alleles of the DNA repair genes XRCC1 (c.839G>A, rs25489; and c.1196A>G, rs25487), XPA (c.-4A>G, rs1800975), and XPD (c.2251A>C, rs13181) and the progression and severity of neoplasias in the urinary bladder and kidney in patients of three distinct ethnic groups, Bashkir, Russians, and Tatar, residing in the Republic of Bashkorostan. The study enrolled 468 cancer patients and 351 healthy individuals. Genotyping for polymorphic alleles was carried out using the PCR-RFLP method. We identified an association between allele A of the c.839G>A locus of the XRCC1 gene and the incidence of the bladder cancer (BC) and renal cell carcinoma (RCC) in the Tatar study group, using the additive genetic effects model (Odds Ratio (OR) = 5.23 and OR = 3.90). In turn, the heterozygous G/A genotype frequency was significantly higher in the RCC patients of Bashkir ethnic origin, compared with the control group (p = 0.0061, OR = 4.72). Additional analysis with consideration of participants smoking status showed that the G/A genotype is significantly more frequent in smokers with BC (OR = 1.96, p = 0.05) than in healthy smokers. We also determined, using the recessive genetic model, that the genotype A/A of the c.1196A>G locus of the XRCC1 gene was associated with a higher risk of BC in the Russian cohort (OR = 2.29, p = 0.0082) and an increased incidence of RCC in the Bashkir group (OR = 4.06, p = 0.05). A similar association was obtained for smokers. In contrast, the allele c.2251A>C in the XPD gene associated with a lower risk for BC and RCC in the Tatars (p = 0.0003, OR = 0.48 and p < 0.0001, OR = 0.37) in the additive model and in the Bashkirs (p = 0.0083, OR = 0.12) and Russians (p = 0.0001, OR = 0.14) in the recessive model. Further, we uncovered that polymorphism c.839G>A in the XRCC1 gene contributes to the progression of noninvasive and invasive BC and promotes RCC at early and advanced stages of the disease. Thus, we identified similar associations between DNA repair gene polymorphisms and the incidence and progression of BC and RCC. We propose that this result points to the involvement of common pathogenetic mechanisms in the initiation and progression of the urinary neoplasias.     

Analysis of the cytochrome P450 1A1 (CYP1A1) gene polymorphism in the ethnic groups of the republic of Bashkortostan

In three ethnic groups from the Republic of Bashkortostan, Russians (N = 451), Tatars (N= 333), and Bashkirs (N = 171), allele, genotype, and haplotype frequency distribution patterns of the CYP1A1 gene single nucleotide polymorphisms, A2455G and T33801C, were investigated. Substantial interethnic differences in the allele frequency distribution patterns of the CYP1A1 polymorphisms A2455G and T3801C (χ ² = 15.61, d.f. = 2, P = 0.0001; and χ ² = 22.10, d.f. = 2, P = 0.0001, respectively) were observed. Pairwise comparison showed that ethnic groups of Tatars and Russians were similar in the A2455G allele frequencies (χ ² = 1.10, d.f. = 1, P = 0.30). However, in case of the T3801C marker, statistically significant differences were revealed (χ ² = 4.56, d.f. = 1, P = 0.032). At the same time, Bashkir ethnic group was found to be statistically significantly different from Russians and Tatars in the CYP1A1 polymorphic allele frequency distribution patterns (χ ² = 15.74, d.f. = 2, P = 0.0001; and χ ² = 7.47, d.f. = 1, P = 0.024, for A2455G, and χ ² = 6.46, d.f. = 1, P = 0.011; and χ ² = 21.36, d.f. = 1, P = 0.0001, for T3801C). Analysis of the CYP1A1 haplotype diversity showed that in terms of the CYP1A1 haplotype frequency distribution patterns, Bashkir ethnic group was statistically significantly different from both Russians (χ ² = 30.07, d.f. = 3, P = 0.0001) and Tatars (χ ² = 11.28, d.f. = 3, P = 0.013). The differences observed were caused by the high frequency of haplotype CYP1A1*2B, which was represented by a combination of rare alleles of the CYP1A1 polymorphisms A2455G and T3801C in Bashkirs (5.81%). On the other hand, the ethnic groups of Russians and Tatars residing in the Republic of Bashkortostan were characterized by similar frequencies of the CYP1A1 haplotypes (χ ² = 6.322, d.f. = 3, P = 0.127). The data obtained could be used in further investigations of the genetic bases of ecology dependant diseases and in the risk groups in the Republic of Bashkortostan.