Changes in erythropoietin and vascular endothelial growth factor following the use of different altitude training concepts.

BACKGROUND: Erythropoietin (EPO) and vascular endothelial growth factor (VEGF) are important factors regulating erythropoiesis and angiogenesis. Altitude/hypoxic training may induce elevated VEGF-A and EPO levels. However, it appears that the range of adaptive changes depends largely on the training method used. Therefore, we investigated the changes in EPO and VEGF-A levels in athletes using three different altitude/hypoxic training concepts. METHODS: Thirty-four male cyclists were randomly divided into four groups: LH-TL group ("live high-train low" protocol), HiHiLo ("live high - base train high - interval train low" procedure), IHT ("intermittent hypoxic training") and control group (CN, normoxic training). The same 4-week training program was used in all groups. Blood samples were taken before and after each training week in order to evaluate serum EPO and VEGF-A levels. RESULTS: In the LH-TL and HiHiLo groups, EPO increased (P<0.001) after 1st week and remained elevated until 3rd week of altitude training. In the IHT and CN groups, EPO did not change significantly. VEGF-A was higher (P<0.001) after 2nd and 3rd week of training in the IHT group. In the HiHiLo group, VEGF-A changed (P<0.05) only after 3rd week. No significant changes of VEGF-A were noted in the LH-TL and CN groups. CONCLUSIONS: Altitude/hypoxic training is effective in increasing VEGF-A and EPO levels. However, a training method plays a key role in the pattern of adaptations. EPO level increase only when an adequate hypoxic dose is provided, whereas VEGF-A increases when the hypoxic exposure is combined with exercise, particularly at high intensity.

Changes in the contents of selected polycyclic aromatic hydrocarbons in soils of various types.

The aim of the paper was to determine the stability and the decomposition intensity of selected polycyclic aromatic hydrocarbons (fluorene, anthracene, pyrene, and chrysene) in soils that are under agricultural use. Soil was sampled from the arable layer that is representative of the Kujawy and Pomorze Provinces, which are located in the northwestern part of Poland. The soil samples were polluted with selected PAHs at an amount corresponding to 10 mg PAHs/kg. PAH-polluted soil samples were incubated for 10, 30, 60, 120, 180, and 360 days at a temperature of 20-25 °C and a fixed moisture of 50% field water capacity. High-performance liquid chromatography (HPLC) was used to determine the content of PAHs. It was found that the process of the degradation of PAHs was most intensive during the first 30 days of the experiment; however, three-ring PAHs (fluorene and anthracene) definitely decomposed faster than the four-ring ones (pyrene and chrysene). The results also confirm the significant role of soil organic matter in sorption and activation processes of PAHs.

Association of adiponectin gene G276T polymorphism with atherogenic indicators in obese children.

Adiponectin plays a protective role against atherosclerosis. Genetic investigation has revealed that G276T adiponectin gene polymorphism is related to adiponectin concentration and metabolic disturbances. The aim of the present study was to investigate the association of adiponectin gene G276T polymorphism with indices of atherosclerosis in obese children. We examined 159 children (125 obese and 34 non-obese). G276T of adiponectin gene polymorphism was identified using a PCR-RFLP method. The intima media thickness (IMT) was evaluated in 82 patients. In all children, the anthropometric indices, fasting plasma total cholesterol (TC), HDL and LDL cholesterol, triglycerydes (TG), C-reactive protein (CRP), and adiponectin were measured. Oral glucose tolerance test (OGTT) also was performed. We found that the obese patients presented with higher values of atherogenic indicators than the non-obese patients. The indicators positively correlated with CRP and lipid concentrations. Ninety one percent of obese children presented with elevated IMT which correlated with CRP. The children with GG genotype (GG + GT allele) had lower values of BMI, TC, and TG but higher adiponectin concentrations. The mean level of adiponectin was statistically decreased in the compared with the homozygous TT children. The other anthropometric and atherogenic indicators did not differ between these two sets of obese children. We conclude that adiponectin concentrations were decreased in children with polymorphism G276T in adiponectin gene. The study, however, failed to show significant associations between carotid IMT, lipid markers, blood pressure, or HOMA-IR in obese children.

Relation of fat-mass and obesity-associated gene polymorphism to fat mass content and body mass index in obese children.

Fat mass content, fat distribution, and fat-mass and obesity associated (FTO) gene have been reported among a broad spectrum of genetic variation connected with body weight. The aim of our study was to investigate whether the T/A rs9939609 polymorphism of the FTO gene may influence obesity and metabolic indices in children. A 160 children were examined (136 obese and 24 non-obese). The anthropometric measurements and calculations included: height, weight, waist and hip circumference, sum of the thickness of 3 and 10 skin folds, % of fat content, % FAT- BIA , % LBM-BIA. BMI, SDS of BMI, WHR, and WHtR. Fasting plasma total cholesterol (TC), HDL and LDL-cholesterol, triglycerides (TG), oral glucose tolerance test (OGTT), and HOMA-IR were analyzed and the blood pressure were measured. The rs9939609 polymorphism of FTO gene was genotyped by allele-specific real-time polymerase chain- reaction (RT-PCR). We found that the mean concentrations of TC, TG, LDLC, and HOMA-IR were significantly higher, and HDL was lower in the obese than in non-obese children. The presence of TT, but not AA alleles, related to the percentage of fat content, BMI, and z-score of BMI. None of the other anthropometric indices did differ between the children with gene polymorphism and wild homozygous. In conclusion, rs9939609 polymorphism in the fat-mass and obesity-associated gene is associated with BMI and the percent of fat content in children.

Association between the polymorphism A/G at position 49 of exon 1 of the CTLA-4 gene and antithyroid antibody production in children with Hashimoto's thyroiditis.

UNLABELLED: CTLA-4 gene is considered to be one of the strongest factors determining the predisposition to antithyroid antibody (Ab) production. The aim of the study was to evaluate the association of the polymorphism A/G of exon 1 of CTLA-4 gene and antithyroid Ab level in children with Hashimoto's thyroiditis (HT). MATERIAL AND METHODS: 45 children with HT (aged 14.9 ± 2, range 8.1-7.9) and 55 healthy controls (aged 14.8 ± 2.34, range 8.0-17.4) were enrolled. Controls were euthyroid and free from any autoimmune disease. CTLA-4 gene (+49)A/G polymorphism was evaluated by a single-strand conformation polymorphism method and restriction fragment-length polymorphism. RESULTS: The frequency of GG genotype in HT children was significantly higher than in controls: 31 vs. 14.5% respectively (p < 0.04, OR = 2.65, CI = 0.99-7.06). Anti-Tg Ab titers were higher in patients homozygous for G allele than with AA genotype. The GG genotype seemed to be protective from hypothyroidism at the moment of HT diagnosis, but this observation was not statistically confirmed. CONCLUSIONS: Our study provides the evidence supporting the association between CTLA-4 gene (+49)A/G polymorphism and the susceptibility to HT in Polish children and confirms the existence of a link between (+49)A/G polymorphism and anti-Tg Ab level.

[The influence of polymorphism the Gly972Arg variant insulin receptor substrate-1 (IRS-1) gene, and G-308A TNF-alpha gene on obesity and insulin resistance in children with obesity]. / Wplyw polimorfizmu Gly972Arg substratu genu receptora insulinowego IRS-1 oraz G-308A genu TNF-alpha na stopien otylosci i zaburzenia gospodarki weglowodanowej u dzieci z otyloscia.

UNLABELLED: Genetic factors play a role in the pathogenesis of insulin resistance in obese subjects. The insulin receptor substrate-1 (IRS-1) and IRS-2 are the most important elements of the insulin-signaling pathways, and mutations in this gene have been reported to play a role in determining insulin resistance, particulary in presence of obesity. The polymorpism of the TNF-a-308 gene is also involved in the development of obesity-related insulin resistance, therefore, we investigated whether the IRS-1 and TNF-a polymorphism can predict conversion to insulin resistance and obesity parameters in children with obesity. MATERIAL AND METHODS: The 70 children with obesity simplex were included in this study (9-18 y.o). The antropometric investigations: weight, height, BMI, SDS for BMI, WHR, sum of 3, 10 skinfolds, and percent of body fat by Slaughter's equation was calculated. In each children after 12 hour overnight fast glucose, insulin, leptin and lipids: triglycerides (Tg), cholesterol total (Chol-T), cholesterol HDL (Chol-HDL), cholesterol LDL (Chol-LDL) were measured. The oral glucose tolerance test was performed and HOMA-IR was calculated. RESULTS: Two variants of genotypic IRS-1 were obtained: C/C(85.7 %), A/C(14.3%), and 3 variants of TNF-a G/G 68 % A/G 29% A/A 3%. Statistical analysis of anthropometric and biochemical variables in groups C/C, vs A/C and variables between IRS and TNF (G/G, A/G + A/A) groups was performed. We did not find any significant differences between these groups in the t-Student test. The girls heterozygous for the A allele--A/C (IRS) had higher body weight than girls who were homozygous C/C (chi(2) =3.87, Pr>chi(2)=0,048). In smaller children studies, both polymorphism--IRS and TNF seems not to be associated with the degree of obesity and insulin resistance.

[The frequency of CTLA-4 gene polymorphism at position 49 exon 1 in children with Hashimoto's thyroiditis]. / Wystepowanie polimorfizmu w pozycji 49 eksonie 1 genu CTLA-4 u dzieci z choroba Hashimoto.

INTRODUCTION: The cytotoxic T lymphocyte antigen-4 (CTLA-4) is a molecule present on T cells that plays a critical role in the down regulation of antigen-activated immune responses. Its gene polymorphism is a strong candidate gene for conferring susceptibility to thyroid autoimmunity. THE AIM of the study was to analyse the frequency of CTLA-4 exon 1 polymorphism (A49G) in children with Hashimoto's thyroiditis. MATERIAL AND METHODS: Blood samples from 68 children were analysed: 30 with Hashimoto's thyroiditis and 38 healthy, age-matched controls, mean age: 13.6 years. CTLA-4 exon 1 polymorphism (A49G) was defined by the PCR method and single-strand conformational polymorphism analysis and confirmed by using BbvI enzyme. Statistical analysis was performed using the t- test and Chi-square test. RESULTS: Polymorphism analysis showed that statistically significant more patients with Hashimoto's thyroiditis were homozygous for G (Ala), and G allele frequency was significant higher than in the control group. CONCLUSION: The results confirmed the association between CTLA-4 exon 1 polymorphism (A49G) and Hashimoto's thyroiditis in polish children.

[Tumor necrosis factor alpha (TNF-alpha) gene G-308A polymorphism relationship to insulin resistance and lipid abnormalities in children with obesity]. / Wplyw polimorfizmu G-308A genu czynnika martwicy guza alfa (TNF-alpha) na zaburzenia gospodarki weglowodanowej i lipidowej u dzieci z otyloscia.

INTRODUCTIONS: High levels of tumor necrosis factor (TNF-alpha)-cytokine produced by adipocytes, are involved in the development of obesity-related insulin resistance and metabolic syndrome. Therefore, we investigated whether the polymorphism of TNF-alpha-308 gene can predict the conversion from insulin resistance and obesity parameters in children with obesity. THE STUDY comprised 72 children with obesity simplex (9-18 y.o). The following anthropometric parameters: weight, height, BMI, SDS for BMI, WHR, sum of 3, 10 skinfolds and percentage of body fat by Slaughter's equation were calculated. In each child, after 12 hour overnight fast, glucose, insulin, leptin and lipids: triglycerides (Tg), cholesterol total (Chol-T), cholesterol HDL (Chol-HDL), cholesterol LDL (Chol-LDL) were measured. The oral glucose tolerance test was performed and HOMA-IR was calculated. RESULTS: Using the technique of PCR-RELP and SSCP, three variants of genotype TNF-alpha were obtained: G/G-68%, A/G-29%, A/A-3%. Statistical analysis of anthropometric, biochemical and leptin variables in groups G/G, vs. A/G +A/A, in boys and girls was performed. We did not find any significant differences between groups (G/G vs. A/G +A/A in all group, and between G/G girls, boys vs A/G +A/A girls, boys) Our data indicate that in smaller children studies, TNF-alpha polymorphism does not seem not to be associated with the degree of obesity and insulin resistance.

Germline mutations 657del5 of the NBS1 gene contribute significantly to the incidence of breast cancer in Central Poland.

Recent studies have demonstrated that heterozygous carriers of the NBS1 657del5 mutation have an increased risk for familial and bilateral breast cancer, but similar studies in consecutive breast cancer patients were inconclusive. Here, in a study of 562 nonselected breast cancer patients from Central Poland, we found 11 (1.96%) 657del5 mutation carriers vs. 3.47 expected (OR 3.21, 95%CI: 1.36-7.61, p = 0.0107) and only 9 (1.6%) carriers of the 5382insC mutation of the BRCA1 gene, most frequently found among breast cancer patients in Poland. No carriers of R215W, another pathogenic mutation of the NBS1 gene, were found in the present study. All carriers of the 657del5 mutation had sporadic breast tumors while 5 of 9 5382insC carriers had a family history of breast/ovarian cancer or bilateral breast carcinoma. In the pooled group of patients from the present and our previous study, carried out also in patients from Central Poland, we obtained the following risk estimates (OR) for 657del5 carriers, as related to the age at breast cancer diagnosis: < 40 years: 8.36; (95%CI: 2.57-27.27) p = 0.0003; < 50 years: 4.27 (95%CI: 1.67-10.89) p = 0.003; > or = 50 years: 2.40 (95%CI: 0.91-6.35) p = 0.1250; all ages: 3.13 (95% CI: 1.40-7.00) p = 0.0066. These findings demonstrate conclusively that NBS1 657del5 mutation carriers have a significantly, though moderately increased, age-related risk of breast cancer, and imply that in populations with a high 657del5 carrier frequency this mutation may contribute substantially to the overall incidence of breast cancer, particularly in younger age groups.

Preliminary studies on DNA retardation by MutS applied to the detection of point mutations in clinical samples.

MutS ability to bind DNA mismatches was applied to the detection of point mutations in PCR products. MutS recognized mismatches from single up to five nucleotides and retarded the electrophoretic migration of mismatched DNA. The electrophoretic detection of insertions/deletions above three nucleotides is also possible without MutS, thanks to the DNA mobility shift caused by the presence of large insertion/deletion loops in the heteroduplex DNA. Thus, the method enables the search for a broad range of mutations: from single up to several nucleotides. The mobility shift assays were carried out in polyacrylamide gels stained with SYBR-Gold. One assay required 50-200 ng of PCR product and 1-3 microg of Thermus thermophilus his6-MutS protein. The advantages of this approach are: the small amounts of DNA required for the examination, simple and fast staining, no demand for PCR product purification, no labelling and radioisotopes required. The method was tested in the detection of cancer predisposing mutations in RET, hMSH2, hMLH1, BRCA1, BRCA2 and NBS1 genes. The approach appears to be promising in screening for unknown point mutations.