Association of Polymorphic Variants of TCF7L2 and PPARG Genes with Metabolic Markers in Patients with Early Disorders of Carbohydrate Metabolism.

We performed complex analysis of the association of polymorphic variants rs7903146 of the TCF7L2 gene and rs1801282 of the PPARG gene with metabolic parameters, insulin resistance, and ß-cell function in a group of patients with early signs of carbohydrate metabolism disturbances in a sample of Tyumen citizens. The study group consisted of 64 people (39 women, 25 men) aged 40-70 years. The distribution of frequencies of alleles and genotypes of the polymorphic markers rs7903146 and rs1801282 was analyzed and associations of carriage of major homozygous polymorphisms with various phenotypic traits were identified. Genotyping for polymorphic variants of TCF7L2 and PPARG genes was performed using allele-specific PCR with primers provided by Synthol company. Carriers of homozygotes for allele C of the polymorphic marker rs7903146 significantly differed from other respondents by a higher level of C-peptide, as well as by the presence of associations with waist circumference, elevated level of glycated hemoglobin, and arterial hypertension. Carriers of homozygotes for the allele C of the rs1801282 polymorphism of the PPARG gene differed from the group of carriers of homozygotes for the allele G and the group of heterozygote carriers by higher levels of triglycerides, as well as the presence of associations with waist circumference and the level of glycated hemoglobin.

MTNR1A and MTNR1B Gene Variants of the Melatonin Receptor and Arterial Stiffness in Persons without Arterial Hypertension.

A comparative analysis of vascular stiffness indices and the results of blood test was carried out in 85 healthy donors aged 19-64 years, carriers of polymorphic variants of type 1 and type 2 melatonin receptor genes. The associations of polymorphic markers of type 1 MTNR1A (rs34532313) and type 2 MTNR1B (rs10830963) melatonin receptor genes with parameters of vascular stiffness and blood parameters in healthy patients were studied. Genotyping was performed using allele-specific PCR. In all patients, 24-h BP monitoring with assessment of arterial stiffness was performed. Allele C homozygotes of MTNR1A differed significantly from carriers of the major T allele by elevated triglyceride, LDL, and fibrinogen levels. The major allele C of the rs10830963 polymorphic variant of the MTNR1B gene is associated with elevated LDL and triglycerides, as well as with individual differences in the elastic properties of the vascular wall in the examined subjects.

Effect of Angiotensin-I Converting Enzyme Gene Insertion/Deletion Polymorphism on genome instability in children living in Russian Arctic.

Main risks of arterial hypertension manifest in childhood. Children living in the Far North are especially susceptible to this. There is a need for an inexpensive, non-invasive and simple diagnosis of the risk of childhood pathologies. It was previously found that the genotype DD of the in/del polymorphic marker of the ACE gene is found in people at risk of developing cardiovascular pathologies. Buccal micronucleus cytome assay and genetic analysis were used in the work. In total, 77 schoolchildren from the city of Apatity, aged 15-17 years old, were examined. We have shown that carriers of the D allele have a tendency to an increase in the frequency of cells with micronuclei. In the case of homozygous I/I variant, the frequency of occurrence of cells with karyopycnosis is significantly higher than in carriers of allele D. Polymorphic marker in/del of the ACE gene is associated with apoptotic changes in the cells of the studied children. The in/del polymorphic marker of the ACE gene can be used as a prognostic marker of the processes of genome destabilization at the early stages of development of the human body.

[Eating behavior, sleep self-assessment and personality traits of students with extreme chronotypes]. / Pishchevoe povedenie, samootsenka sna i osobennosti emotsional'noi sfery lichnosti u studentov krainikh khronotipov.

OBJECTIVE: To identify the differences in eating behavior, sleep self-assessment and personality traits in students with extreme chronotypes. MATERIAL AND METHODS: Students (n=350), aged 17 to 23 years, with extreme chronotypes (owls/larks) were studied. Eating behavior was assessed with the Three-Factor Eating Questionnaire (TFEQ-R18). Evaluation of the condition of the cycle sleep-wakefulness (sleep quality, sleep hygiene, daytime sleepiness) and psychophysiological differences (Eysenck Personality Questionnaire, Spielberger-Khanin anxiety inventory and the Taylor Manifest Anxiety Scale (TMAS); the Stress test) was performed. RESULTS AND CONCLUSION: Higher neuroticism, psychoticism and anxiety, low resistance to stress, reduced quality and hygiene of sleep with increased daytime sleepiness are found in people with evening preferences in comparison with the morning chronotype. Eating behavior according to TFEQ-R18 is characterized by a lack of cognitive restraint. The results may indicate preclinical health disorders, which are affected by lifestyle factors. The number of alcohol consumers and smokers is two and more than three times higher, respectively, among students with evening chronotype. The changes identified in people with evening chronotype require additional attention to student's health.

Psychometric properties of the Russian version of the Pediatric Daytime Sleepiness Scale (PDSS).

Insufficient sleep could severely impair both cognitive and learning skills. More prominent changes are found in children and adolescents. Tools used to estimate sleepiness in the adult population are commonly inappropriate for children. The objective of our study was to provide a reliable instrument to measure excessive sleepiness for upcoming studies in Russian-speaking children, applying the Russian version of Pediatric Daytime Sleepiness Scale (PDSS). The following tasks were resolved in our study: translation, validation, and analysis of psychometric properties of the Russian adaptation of the PDSS by standard tests. After the semantic validation of the instrument through a multi-stage translation process we checked its psychometric validation. A total of 552 students, consisting of N = 285 for the exploratory factor analysis (EFA), N = 267 for the confirmatory factor analysis (CFA) and N = 204 for test-retest analysis of public elementary schools located in Northern Russia completed the PDSS and Munich Chronotype Questionnaire to estimate sleep parameters in the classroom during the lessons. Response rate was 90%; excluded cases contained no data. Further, 204 of our participants completed the PDSS in a 3 months interval to check the test-retest reliability. Internal consistency was measured by Cronbach's alpha coefficients and CFA was used to test factorial validity of the tool. Concurrent validity and test-retest reliability were assessed via intra-class coefficient. Internal consistency of the PDSS scale was high (Cronbach's α = 0.8). The construct validity of the PDSS was supported by CFA (factor loadings were from 0.438 to 0.727) and the test-retest reliability demonstrated by the intra-class coefficient was 0.70. The total PDSS score was independent of sex. The mean total value of PDSS was 11.95 ± 6.24. Higher scores on PDSS were negatively correlated with sleep duration. Thus, the construct validity of the instrument remains valid and could be used for Russian-speaking youth samples in the evaluation of daytime sleepiness. It could be useful in future applications by sleep scientists and health practitioners.

[Indices of cell-mediated immunity in rheumatoid arthritis: the role of cytomegalovirus infection.]

The pathogenesis of rheumatoid arthritis (RA) is driven by a combined action of genetic and environmental factors, which can upset the balance between the effector and regulatory components of the immune system. An important actor in maintaining such balance is T cells, especially regulatory T lymphocytes (Treg), but the mechanisms behind the functioning of T cell subpopulations and the roles of individual etiological factors in RA have not been fully elucidated. This study aimed to investigate the indices of cell-mediated immunity, especially T- and Treg cells, in RA patients depending on the disease activity and presence of cytomegalovirus (CMV) infection. The expression of membrane and intracellular molecular markers of lymphocytes was estimated by multicolor flow cytometry. The content of antibodies to CMV in blood plasma was measured by enzyme immunoassays. Patients with RA had reliably reduced numbers of cells with the phenotypes CD4+FOXP3+, CD4+CD25+FOXP3+ correlating with the stage of RA activity. RA patients with CMV infection showed a reduction in the number of regulatory T cells (Treg), CD3+ T lymphocytes, CD3+CD8+ cells in peripheral blood. At the same time, RA involved a rise in the level of B cells and CD4+CD25+ Т cells. The level of antibodies to CMV was observed to grow in line with RA activity. Thus, the data obtained suggest that the presence of CMV infection can significantly influence the state of individual lymphocyte subpopulations during RA development.

[Sleep quality and its parameters in schoolchildren]. / Kachestvo i parametry sna u shkol'nikov.

AIM: Sleep disorders are very common among teenagers. One of possible reasons is a discrepancy between social and biological activity of children. It is known that adolescents with a late chronotype (e.g., a later midpoint of sleep) have a lowered academic achievement. The aim of our study was to evaluate sleep quality and other sleep-wake characteristics in relation to academic scores self-reported by teenagers living in urban and rural areas of Republic of Karelia. MATERIAL AND METHODS: Five hundred respondents, aged 10-17 years, participated in a questionnaire survey of chronotype and sleep quality. The Munich chronotype questionnaire (MCTQ) and the Pittsburgh Sleep Quality Index (PSQI) were used. Sleep duration during week and weekend, sleep quality and hygiene were assessed. RESULTS AND CONCLUSION: A decrease in sleep duration in adolescents, aged 10-18 years, was observed. A larger social jetlag was significantly higher in girls compared to boys. Gender had a significant impact on sleep quality. More than 10% of the total variation in academic achievement of Karelian teenagers can be explained by the difference in their gender and average sleep duration.

[Circadian CLOCK Gene Variants, Parameters of Arterial Stiffness and Blood Pressure Levels in Subjects Without Arterial Hypertension].

CLOCK gene polymorphic variants, parameters of arterial stiffness and blood pressure (BP) variability were studied in 115 normotensive Russian patients without cardiovascular diseases (62 men, 53 women; mean age 36.4+/-1.01 years). Examination included ECG, 24h BP monitoring, duplex scan of carotid arteries, registration of vascular stiffness parameters, and genotyping of the following polymorphic markers of CLOCK: 3111T>C (3-untranslated region), 862T>C (exon 9) and 257T>G (promoter region) by PCR-RFLP method. Pulse wave velocity was not significantly different among carriers of various 257T>G, 862T>C, 3111T>C genotypes. Augmentation index (Aix night, Aix max) values were lower in individuals with genotypes allegedly associated with essential hypertension and ischemic heart disease (257GG, 862CC, 3111CC). Arterial stiffness index (ASI) was significantly higher in men having CC genotype of 862T>C. Polymorphic variants 257T>G and 862T>C SNPs of CLOCK gene were found to be associated with parameters reflecting BP variability (morning rise of diastolic BP and rate of BP rise). These results provide a basis for suggestion that the CLOCK gene polymorphism is one of factors determining elastic properties of vascular wall. The suggestion is supported by discussion of possible molecular mechanisms of circadian genes impact on elasticity and rigidity of vessel.

Expression of circadian rhythm genes CLOCK, BMAL1, and PER1 in buccal epithelial cells of patients with essential arterial hypertension in dependence on polymorphic variants of CLOCK and BMAL1 genes.

The transcript levels of circadian rhythm genes CLOCK, BMAL1, and PER1 in buccal epithelial cells of the patients with essential arterial hypertension was analyzed in relation to polymorphic variants of CLOCK and BMAL1 genes. These levels were assessed with realtime PCR method at daily hours 9, 13, and 17. The significant differences were revealed in transcript levels of the examined genes in patients with various genotypes at the polymorphic markers 3111TC and 257TG regulatory regions of CLOCK gene. The study detected no significant differences among the carriers of various genotypes at polymorphic markers 862TC and 2121GA of CLOCK gene and 56445TC of BMAL1 gene.

Expression of circadian Per1 and Per2 genes in the liver and breast tumor tissues of HER2/neu transgenic mice of different age.

The expression of Per1, Per2, and Cry1 circadian genes in the liver and breast tumors were studied by real-time PCR in FVB/N mice of different age transfected with HER-2/neu gene. The expression of Per1 and Per2 genes in breast tumor tissue decreased in comparison with their expression in the lever. The expression of these genes decreased with age in both the liver and tumor tissue.

[Association of T3111C polymorphism in 3'-untranslated region of the CLOCK gene with the risk of essential arterial hypertension and coronary artery disease in the Russian population Karelia].

Allele and genotype distributions of the T3111C polymorphism in 3'-untranslated region of the CLOCKgene were examined in the groups of Russian patients with essential arterial hypertension (EAH) and coronary artery disease (CAD), and in control group of Russia residents of the Republic of Karelia. The genotype frequency distributions of the polymorphism examined in the EAH and CAD patients were statistically significantly different from that in the individuals without clinical signs of these diseases. The CC genotype frequency in EAH and CAD males was higher, and in the corresponding females it was lower than in males and females from the control group. Male CC carriers were characterized by a possible increased risk of EAH: OR (95% CI) = 1.42 (0.56; 3.58). Moreover, the presence of the CC genotype in males could increase the risk of CAD: OR (95% CI) = 1.58 (0.63; 3.93).

[Transfection of eucaryotic cells using cytochrome CYP450 2B6 gene: sensitivity to cyclophosphamide]. / Transfektsiia éukarioticheskikh kletok genom tsitokhroma CYP450 2B6: poiavlenie chuvstvitel'nosti k tsiklofosfamidu.

Sensitivity of 293 human epithelial kidney cells transfected by human cytochrome CYP450 gene to cyclophosphamide was investigated. Transfection was carried out by plasmid DNA containing CYP2B6 gene complexed with cationic liposomes. Liposomes were prepared from mixture of cationic lipids and cholesterol at different molar ratios. Experimental protocol included the following steps: transfection of epithelial kidney cells by complexes of plasmid DNA-cationic liposomes, clone selection in the medium with antibiotic Geneticin G418, selected clone harvesting and their treatment by cyclophosphamide as following cytotoxicity evaluation. It was shown that addition of 0.25 mM of cyclophosphamide resulted in death of 40-45% transfected cell population.

Liposomes containing dopamine entrapped in response to transmembrane ammonium sulfate gradient as carrier system for dopamine delivery into the brain of parkinsonian mice.

The active loading of liposomes with dopamine in response to an ammonium sulfate gradient was studied. This method can be regarded as a mean to more efficiently improve the liposomal dopamine/lipids ratio in comparison to conventional methods of liposome preparation. Trapping efficiency of dopamine into liposomes exhibiting a transmembrane ammonium sulfate gradient was shown to be dependent on liposome lipid composition, lipid concentration and temperature. Dopamine-containing liposomes with alpha-tocopherol in the lipid bilayer were shown to be stable at least for three weeks. It has been found that intraperitoneal (i.p.) administration of conventionally prepared dopamine-containing liposomes as well as liposomes with increased dopamine/lipid ratio may efficiently suppress the expression of parkinsonian symptoms in C57BL/6 mice with experimental parkinsonian syndrome. On the other hand, only through increasing of liposomal dopamine/lipid ratio the complete compensation of dopamine deficiency in the mice brain was achieved. The obtained data may be considered as biochemical evidence in favor of liposomes' ability to act as a carrier system for the delivery of dopamine into the brain.

[Cloning of the human cytochrome p450 2B6 gene in Escherichia coli cells]. / Klonirovanie gena tsitokhroma p450 2B6 cheloveka v kletkakh Escherichia coli.

Human cytochrome P450 2B6 gene from plasmid pUC9, carrying cytochrome CYP2B6 cDNA was cloned into eucariotic expression vector pcDNA 3.1 (+). Cytochrome P450 2B6 gene in recombinant plasmid pcDNA 3.1 (+)/CYP 2B6 was expressed in E. coli cells. The expression of catalytically active recombinant protein was 60-100 nm per liter of the culture medium.

[Concentrating dopamine in liposomes using transmembrane gradient of ammonium sulfate]. / Kontsentirovanie dopamina v liposomakh s pomoshch'iu transmembrannogo gradienta kontsentratsii sul'fata ammoniia.

The active loading of liposomes from egg phosphatidylcholine and cholesterol with dopamine using an ammonium sulfate gradient was studied. Our conditions allowed the enrichment of the monolamellar liposomes with 90% dopamine added to the medium and, thus, considerably improve the dopamine/lipid ratio. Dopamine-containing liposomes with tocopherol in their lipid bilayer were shown to be stable for 3 weeks.

[Preparation and certain properties of the liposomal substance 2,4-di(1-methyl-3-hydroxybutyl)deuteroporphyrin-IX]. / Poluchenie i nekotorye svoistva liposomnogo preparata 2,4-di(1-metil-3-gidroksibutil)deiteroporfirina-IX.

A liposome preparation of a porphyrin photosensitizer for photodynamic therapy of tumors (PDT) was obtained. The in vitro efficiency of the photosensitizer was enhanced 2.5-fold through the liposome formulation. The composition and some properties of the new preparation were studied. An algorithm for a complex approach to the prediction of photosensitizer efficiencies by model experiments in vitro was developed. This approach is based on the use of two models: the determination of coefficient of distribution between n-octanol and a phosphate buffer, pH 7.4, and the determination of the cytotoxic effect on the culture of CaOv ovarian adenocarcinoma cells.