Restriction Fragment Length Polymorphism-polymerase Chain Research Articles

CARD15 Gene Polymorphisms Are Associated with Tuberculosis Susceptibility in Chinese Holstein Cows.

Bovine tuberculosis (BTB) is a significant veterinary and financial problem in many parts of the world. Associations between specific host genes and susceptibility to mycobacterial infections, such as tuberculosis, have been reported in several species. The objective of this study was to identify and evaluate the relationship of single-nucleotide polymorphisms (SNPs) in the CARD15 gene with susceptibility to BTB in Chinese Holstein cows. DNA samples from 201 Chinese Holstein cows (103 cases and 98 controls) were collected from Kunming City, Yuxi City, and Dali City in China. SNPs in the CARD15 gene were assessed using polymerase chain reaction (PCR) and restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). Case-control association testing and statistical analysis identified six SNPs associated with susceptibility to BTB in Chinese Holstein cows. The frequency of genotypes C/T, A/G, A/G, A/G, C/T, and A/G in E4 (-37), 208, 1644, 1648, 1799, and E10 (+107), respectively, was significantly higher in cases than in controls, and also the alleles C, A, A, G, T, and A, respectively, were associated with a greater relative risk in cases than in controls. The distribution of two haplotypes, TGGACA and CAGACA, was significantly different between cases and controls. Overall, this case-control study suggested that E4 (-37)(C/T), 208(A/G), 1644(A/G), 1648(A/G), 1799(C/T), and E10 (+107)(A/G) in the CARD15 gene were significantly associated with susceptibility to BTB in Chinese Holstein cows and that haplotypes TGGACA and CAGACA could be used as genetic markers in marker-assisted breeding programs for breeding cows with high resistance to BTB.

Mutation analysis of the phenylalanine hydroxylase gene in Azerbaijani population, a report from West Azerbaijan province of Iran.

Phenylketonuria (PKU) is a genetic inborn error of phenylalanine (Phe) metabolism resulting from insufficiency in the hepatic enzyme, phenylalanine hydroxylase (PAH), which leads to elevated levels of Phe in the blood. The present study was carried out for mutation analysis of the PAH gene in West Azerbaijan province of Iran. A total of 218 alleles from 40 PKU families were studied using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method. The frequencies of IVS10-11, S67P, R261Q, R252W, IVS11nt-1 g>c, R408Q, and Q232Q mutations were 28(35), 17(21.25), 15(18.75), 3(3.75), 3(3.75), 2(2.5), and 1(1.25), in cases group, and 51(23.4), 31(14.2), 27(12.4), 6(2.75), 6(2.75), 4(1.83), and 2(0.92) in total group, respectively. The mutations of R243Q, 364delG, L333F, 261X, I65T, and R408W were not detected in our samples. It can be concluded that the IVS10-11 mutation has the highest frequency in the tested population. To our knowledge, this report is the first in its own kind and provides better understanding of the genetic heterogeneity, the origin and distributions of PAH mutations in West Azerbaijan province of Iran.

Genetic Effects of eNOS Polymorphisms on Biomarkers Related to Cardiovascular Status in a Population Coexposed to Methylmercury and Lead.

The aim of the present study was to evaluate possible effects of endothelial nitric oxide synthase (eNOS) polymorphisms on systolic (SBP) and diastolic blood pressure (DBP) and on nitrite levels in plasma (NitP) in a population coexposed to methylhemoglobin (MeHg) and lead (Pb) in the Amazonian region, Brazil. Plasmatic levels of hemoglobin Hg (HgP) and Pb (PbP) were determined by inductively coupled plasma-mass spectrometry, whereas NitP were quantified by chemiluminescence. Genotyping was performed by conventional and restriction fragment length polymorphism-polymerase chain reaction assay. The population age ranged from 18 to 87years (mean 40±16), and the distribution between the sexes was homogenous (63 men and 50 women). Mean HgP and PbP were 7.1±6.1 and 1.1±1.1µgL(-1), respectively. PbP was correlated to SBP and DBP, whereas no effects were observed for HgP on blood pressure. Subjects carrying the 4b allele in intron 4 presented greater SBP and DBP compared with those who had the 4a4a genotype. In addition, interactions between alcohol consumption and the -786T/C polymorphism were observed on NitP, i.e., individuals carrying the polymorphic allele and drinkers had lower NitP. Taken together, our data give new insights concerning the genetic effects of eNOS polymorphisms on biomarkers related to cardiovascular status in populations coexposed to Hg and Pb.

CD38 and Interleukin 6 Gene Polymorphism in Egyptians with Diffuse Large B-Cell Lymphoma (DLBCL)

Given the importance of understanding the genetic variations involved in the pathogenesis of non-Hodgkin’s lymphoma (NHL), this pilot study was designed to investigate the impact of CD38 (184C/G; rs6449182) and IL-6 (−174 G/C; rs1800795) gene polymorphism on susceptibility of Egyptians to diffuse large B cell lymphoma (DLBCL); major types of NHL. To the best of our knowledge, this study is the first one that examines CD38 polymorphism in the NHL. Genotyping polymorphism is performed using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) for CD38 and Mutagenically separated PCR (MS-PCR) for IL-6 in 100 Egyptian NHL patients with DLBCL subtype and 119 normal controls. The serum level of IL-6 was measured using Enzyme-linked immunosorbent assay (ELISA). CD38 (184C/G) genotype is significantly increased in NHL patients (p < 0.01), while the GG genotype is significantly increased in controls (p < 0.05). Only two genotypes were found (GG and GC) in IL-6 (−174), no CC in our NHL patients and only one case in the controls. Insignificant change in IL-6 (−174 G/C) genotypes was recorded. Significantly increased serum IL-6 (p < 0.05) was positively correlated (r = 0.17; p < 0.05) with the disease. Taken together, our data stressed the importance of CD38 gene polymorphism in developing DLBCL. Our pilot study indicates that CD38 (184) CG genotype might play a role in DLBCL susceptibility in Egyptians. Additional prospective studies on larger population are needed to confirm our findings.

Candidate genes for carcass traits in a tropical-adapted Brazilian composite beef breed

Backfat thickness (BFT) and ribeye area (REA) are important production traits but, because they are measured late in the animal's life, they have not been efficiently included in breeding programs. The aim of this study was to evaluate whether single nucleotide polymorphisms (SNPs) mapped to the leptin, PPARGC1A, PSMC1, CRH, and FABP4 genes, which influence BFT and REA in Canchim cattle, a composite beef breed (5/8 Charolais + 3/8 Zebu). BFT and REA phenotypic records were obtained by ultrasound measurements from 18-month-old animals. All SNP markers were genotyped by restriction fragment length polymorphism-polymerase chain reaction. Restricted maximum likelihood analysis revealed that the non-synonymous SNP located in exon 2 of the FABP4 gene has an additive effect on BFT (P ≤ 0.05). Significant allele substitution effects showed that the substitution of G by A may lead to a decrease of 0.1055 mm in mean BFT. This information can be used for inclusion of this trait-associated marker in commercial SNP panels.

Mn-SOD 47 CC genotype in combination with high tea consumption may prevent complications in Tunisian type-2 diabetes.

Reactive oxygen species metabolizing enzymes may play an important role in the prevention of type-2 diabetes (T2D) complications. We analyzed the association between Cu/Zn-SOD +35 A/C, Mn-SOD T47C, and CAT -21 A/T gene polymorphisms and complications, in combination with tea consumption in Tunisian T2D. A sample of 366 T2D subjects was enrolled in this study. All participants were asked about tea consumption and frequency. Anthropometric, clinical, and routine biochemical characteristics were obtained from subjects' updated medical records. Malondialdehyde, as an early marker of lipid peroxidation, was measured in plasma samples. Urinary polyphenol derivatives (UPDs), as a marker of polyphenols intake, were assessed by the Folin-Ciocalteu assay. SODs and CAT genotypes were determined by conventional restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) methods. From all subjects, the results showed that in high tea consumers (>3 cups/day), the frequency of the Mn-SOD 47 CC genotype was significantly higher in T2D without complications compared with T2D with complications (P = 0.03; OR = 0.284; 95%CI = 0.086-0.939). However, no significant associations were observed with Cu/Zn-SOD +35 A/C or CAT -21 A/T genes polymorphisms. Additionally, the evaluation of UPDs showed that individuals carrying the Mn-SOD 47 CC genotype and consuming more than three cups of tea per day present significantly higher UPDs (P = 0.038). In conclusion, the Mn-SOD 47 C variant in combination with high tea consumption may provide protection against complications in T2D.

Correlation of multidrug resistance genes and clinical risk factors with glucocorticoid response in patients with inflammatory bowel disease

Objective To investigate the correlation of multidrug resistance gene 1 (MDR1),NR3C1 gene polymorphisms and clinical risk factors with efficacy,dependence,and resistance of glucocorticoid (GC) in patients with inflammatory bowel disease (IBD).Methods Anti coagulation blood samples of 196 healthy controls and 105 IBD patients received GC therapy were collected.There were 62 ulcerative colitis (UC) and 43 Crohn's disease (CD) in the IBD patients.The number of GC sensitive,GC dependent and GC resistant of UC patients were 36,13 and 13,respectively,and those of CD patients were 24,11 and eight.GC refractoriness included GC dependence and resistance.The genotype of MDR1 C3435T and NR3C1 Bcl Ⅰ of all the subjects was detected by the restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR).The correlation between each genotype frequency,clinical features of patients with IBD and the efficacy of GC treatment was analyzed by Chisquare test,Fisher exact probability method or t test.Results Among UC patients,the disease course of GC refractory group and GC resistant group was longer than that of GC sensitive group ((6.660±1.523)years,(6.500±1.111) yearsvs (3.350±0.697) years,t=2.211,P=0.031; t=2.930,P=0.005).The serum level of C reaction protein (CRP) of GC refractory group was higher than that of GC sensitive group ((47.628±13.913) mg/Lvs (16.854±4.121) mg/L,t=2.121,P=0.047).The chronic relapse type was more common in GC refractory UC patients (Fisher exact probability method,P=0.035),and severe patients were more common in UC with GC resistance (Fisher exact probability method,P=0.021).The white blood cell count of GC resistant and GC refractory CD patient was lower than that of GC sensitive CD patients ((5.710 ± 0.604) ×109/L,(5.878±0.405) × 109/L vs (7.814 ±0.670) × 109/L,t=2.334,P=0.028; t=2.045,P=0.018).Patients with extraqntestinal manifestations was more common in CD with GC resistance (Fisher exact probability method,P=0.035).There was no statistically significant difference in the frequencies of MDR1 C3435T,NR3C1 Bcl Ⅰ genotypes,allelic genes and gene carrier among control group and GC sensitive dependent and resistant group of IBD patients.However,the frequency of MDR1 C3435T gene carrier was significantly different between GC sensitive group and GC refractory group,especially between GC sensitive group and GC resistance group (68.33％ vs 48.89％,x2 =4.051,P=0.044; 68.33％ vs 42.86％,x2 =4.274,P =0.039).Conclusions GC sensitivity of IBD patients with MDR1 C3435T loci T gene carrier was higher than that of IBD patients without T gene carrier.NR3C1 gene polymorphisms was not related with GC resistance and GC dependence.Compared with GC sensitive IBD patients,in GC resistant and GC dependent IBD pantient UC patients with long disease course,chronic relapse type,severe type,high level of CRP and CD patients with low white blood cell count and extra-intestinal manifestations were more common. Key words: Inflammatory bowel diseases; MDR1 gene; Receptors, glucocorticoid; Polymorphism,genetic

Detection of high levels of anti-α-galactosyl antibodies in sera of patients with Old World cutaneous leishmaniasis: a possible tool for diagnosis and biomarker for cure in an elimination setting.

In the Kingdom of Saudi Arabia (KSA), Old World cutaneous leishmaniasis (CL) is mainly caused by Leishmania major and Leishmania tropica parasites. Diagnosis of CL is predominately made by clinicians, who at times fail to detect the disease and are unable to identify parasite species. Here, we report the development of a chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) to measure the levels of anti-α-galactosyl antibodies in human sera. Using this assay, we have found that individuals infected with either Leishmania spp. had significantly elevated levels (up to 9-fold higher) of anti-α-Gal IgG compared to healthy control individuals. The assay sensitivity was 96% for L. major (95% CI; 94-98%) and 91% for L. tropica (95% CI; 86-98%) infections and therefore equivalent to restriction fragment length polymorphism-polymerase chain reaction analysis of parasite ITS1 gene. In addition, the assay had higher sensitivity than microscopy analysis, which only detected 68 and 45% of the L. major and L. tropica infections, respectively. Interestingly, up to 2 years following confirmed CL cure individuals had 28-fold higher levels of anti-α-Gal IgG compared to healthy volunteers. Monitoring levels of anti-α-Gal antibodies can be exploited as both a diagnostic tool and as a biomarker of cure of Old World CL in disease elimination settings.

Enzymatic and molecular characteristics ofGeotrichum candidumstrains as a starter culture for malting

Geotrichum candidum yeasts are proposed as a starter culture during malting. They have a double positive effect on the process. They eliminate the fungal pathogenic microflora and improve the technological properties of the finished product, malt. Among published research little or no information can be found considering the biosynthesis of hydrolytic enzymes. Most of the data is related to the production of lipases, proteases and cellulases. This paper examines the enzymology of a number of G. candidum strains. The main focus within the evaluation process was placed on whole cell barley grain cultivation. Hydrolase, which is present in cell wall degradation, was found at a satisfactory level. All tested strains produced both β-1,3-glucanases and β-1,4-glucanases capable of hydrolysing barley β-glucan and reducing the amount of this polysaccharide in the wort. Molecular analysis of the tested strains with the use of restriction fragment length polymorphism–polymerase chain reaction and randomly amplified polymorphic DNA confirmed not only the species affiliation, but also the genetic similarity between the tested strains. Copyright © 2014 The Institute of Brewing & Distilling

Role of the VDR Bsm I and Apa I Polymorphisms in the Risk of Colorectal Cancer in Kashmir

Objective: A case-control study aiming to evaluate the relationship between Bsm I and Apa I restriction fragment gene polymorphisms and colorectal cancer (CRC) was carried out in Kashmir, including a total of 368 subjects (180 cases and 188 controls). Methods: DNA samples extracted from the blood of the subjects were analyzed for 3' untranslated region (3' UTR) Apa I and Bsm I polymorphisms using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). Results: A statistically significant 2.7-fold increased risk was observed in individuals found homozygous for the presence of the ‘b' allele, in comparison to subjects homozygous for the ‘B' allele (odds ratio (OR) 2.7, 95% confidence interval (CI) 1.49-4.86 (Bsm I)), and a statistically insignificant 2-fold increased risk was found among individuals with the ‘aa' genotype, as compared to subjects with the ‘AA' genotype (OR 2.017, 95% CI 0.86-4.7). Our study also yielded statistically significant results when the Apa I polymorphism was stratified by age (≤ 50 years) and dwelling area (rural area), and the Bsm I polymorphism by gender (male gender), suggesting a possible role of Apa I and Bsm I polymorphisms in the etiology of CRC in Kashmir. Conclusion: We conclude that Apa I and Bsm I single-nucleotide polymorphisms (SNPs) in the vitamin D receptor gene (VDR) might be associated with susceptibility to CRC among Kashmiris.

Lack of association between autonomously functioning thyroid nodules and germline polymorphisms of the thyrotropin receptor and Gαs genes in a mild to moderate iodine-deficient Caucasian population.

Mutations of the thyrotropin receptor (TSHR) and/or Gαs gene have been found in a number of, but not all, autonomously functioning thyroid nodules (AFTNs). Recently, in a 15-year-old girl with a hyperfunctioning papillary thyroid carcinoma, we found two somatic and germline single nucleotide polymorphisms (SNPs): a SNP of the TSHR gene (exon 7, codon 187) and a SNP of Gαs gene (exon 8, codon 185). The same silent SNP of the TSHR gene had been reported in patients with AFTN or familial non-autoimmune hyperthyroidism. No further data about the prevalence of the two SNPs in AFTNs as well as in the general population are available in the literature. To clarify the possible role of these SNPs in predisposing to AFTN. Germline DNA was extracted from blood leukocytes of 115 patients with AFTNs (43 males and 72 females, aged 31-85 years, mean ± SD = 64 ± 13) and 100 sex-matched healthy individuals from the same geographic area, which is marginally iodine deficient. The genotype distribution of the two SNPs was investigated by restriction fragment length polymorphism-polymerase chain reaction. The prevalence of the two SNPs in our study population was low and not different to that found in healthy individuals: 8 % of patients vs. 9 % of controls were heterozygous for the TSHR SNP and 4 % patients vs. 6 % controls were heterozygous for the Gαs SNP. One patient harbored both SNPs. These results suggest that these two SNPs do not confer susceptibility for the development of AFTN.

Pharmacogenetics of uridine diphosphate glucuronosyltransferase (UGT2B7) genetic polymorphism on valproic acid pharmacokinetics in epilepsy

Background Sodium valproate is a widely prescribed broad-spectrum antiepileptic drug. It shows high inter-individual variability in pharmacokinetics and pharmacodynamics and has a narrow therapeutic range [1]. We evaluated the effects of polymorphic Uridine diphosphate glucuronosyltransferase (UGT2B7) metabolizing enzyme on the pharmacokinetics of sodium valproate in the patients with epilepsy who showed toxicity to therapy. Materials and methods Genotype analysis of the patients was made with polymerase chain–restriction fragment length polymorphism (RFLP) with sequencing. Plasma drug concentrations were measured with reversed phase high-performance liquid chromatography (HPLC) and concentration–time data were analyzed by using a non-compartmental approach. Results The results of this study suggested a significant genotypic as well as allelic association with valproic acid toxicity for UGT2B7 polymorphic enzymes. The elimination half-life (t1/2=42.2 h) of valproic acid was longer and the clearance rate (CL=947 ml/h) was lower in the poor metabolizers group of UGT2B7 polymorphism who showed toxicity than in the intermediate metabolizers group (t1/2 = 36.5 h, CL = 1,042 ml/h) or the extensive metabolizers group (t1/2 = 27. h, CL = 1,602 ml/h).Conclusions Our findings suggest that the UGT2B7 genetic polymorphism plays a significant role in the steady state concentration of valproic acid, and it thereby has an impact on the toxicity of the valproic acid used in the patients with epilepsy.

Molecular mechanisms of quinolone, macrolide, and tetracycline resistance among Campylobacter isolates from initial stages of broiler production

The aim of this study was to investigate the resistance mechanisms of quinolones, macrolides and tetracycline in campylobacter isolates from grandparent and parent broiler breeders in Spain. Twenty-six isolates were investigated for quinolone resistance, three isolates for macrolide resistance and 39 for tetracycline resistance. All of the quinolone-resistant isolates possessed the mutation Thr86Ile in the quinolone resistance-determining region of gyrA and one isolate possessed the mutation Pro104Ser. Only one Campylobacter coli population (defined by restriction fragment length polymorphism–polymerase chain reaction of flaA and pulsed field gel electrophoresis) was resistant to erythromycin, and the mutation A2075G (23S rDNA) was responsible for macrolide resistance. The tetO gene was found in all of the tetracycline-resistant isolates. Twenty-two out of the 39 isolates investigated by Southern blot possessed chromosomic location of tetO and 17 were located on plasmids. Most of the plasmids with tetO were of around 60 kb and conjugation was demonstrated in a selection of them. In conclusion, we showed that Thr86Ile is highly prevalent in quinolone-resistant isolates as well as mutation A2075G in macrolide-resistant isolates of poultry origin. More variability was found for tetO. The possibility of horizontal transmission of tetO among campylobacter isolates is also an issue of concern in public health.

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BackgroundSickle cell anemia is a chronic inflammatory disease characterized by an increased production of proinflammatory cytokines including tumor necrosis factor-alpha. Hydroxyurea, by decreasing the polymerization of hemoglobin, reduces inflammatory states. The effect of the genetic polymorphisms of sickle cell patients on tumor necrosis factor-alpha levels remains unknown.ObjectiveThe aim of this study was to investigate the association of tumor necrosis factor-alpha levels with β-globin haplotypes and the use of hydroxyurea.MethodsA cross-sectional study was performed of 67 patients with sickle cell anemia diagnosed at steady-state in a referral hospital in Fortaleza, Ceará, Brazil. A group of 26 healthy individuals was used as control. βS haplotype analysis was performed by restriction fragment length polymorphism-polymerase chain reaction. The tumor necrosis factor- alpha levels were measured by the enzyme-linked immunosorbent assay test. Laboratory data (complete blood count and fetal hemoglobin) and information regarding the use of hydroxyurea were obtained from medical records. Statistical analysis was performed using R software with the Kruskal-Wallis and Mann–Whitney tests. Statistical significance was established for p-values < 0.05 for all analyses.ResultsThe mean age of the participants was 35.48 years. Patients with sickle cell anemia had significantly higher tumor necrosis factor-alpha levels than controls (p-values < 0.0001). Tumor necrosis factor-alpha levels were lower in sickle cell anemia patients who were receiving hydroxyurea treatment than those who were not (p-value = 0.1249). Sickle cell anemia patients with Bantu/n genotype had significantly higher levels than patients with the Bantu/Benin genotype (p-value = 0.0021).ConclusionIn summary, βS-globin haplotypes, but not hydroxyurea therapy, have a role in modulating tumor necrosis factor-alpha levels in sickle cell anemia adults at steady-state. Many previous studies have investigated prognosis and inflammatory states in sickle cell anemia patients, but the discovery that tumor necrosis factor-alpha levels vary according to the genetic polymorphism of the patient is a new finding. © 2014 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. All rights reserved.

Atypical Presentation of PKDL due to Leishmania infantum in an HIV-Infected Patient with Relapsing Visceral Leishmaniasis.

We describe the case of an Italian patient with HIV infection who developed an atypical rash resembling post-kala-azar dermal leishmaniasis (PKDL) when receiving liposomal Amphotericin B (L-AMB) for secondary prophylaxis of visceral leishmaniasis (VL). At the time of PKDL appearance, the patient was virologically suppressed but had failed to restore an adequate CD4+ T-cell count. Histology of skin lesions revealed the presence of a granulomatous infiltrate, with lymphocytes, plasma cells, and macrophages, most of which contained Leishmania amastigotes. Restriction fragment length polymorphism-polymerase chain reaction was positive for Leishmania infantum. Paradoxically, cutaneous lesions markedly improved when a new relapse of VL occurred. The patient received meglumine antimoniate, with a rapid clinical response and complete disappearance of cutaneous rash. Unfortunately, the patient had several relapses of VL over the following years, though the interval between them has become wider after restarting maintenance therapy with L-AMB 4 mg/kg/day once a month. Even if rare, PKDL due to Leishmania infantum may occur in Western countries and represents a diagnostic and therapeutic challenge for physicians. The therapeutic management of both PKDL and VL in HIV infection is challenging, because relapses are frequent and evidence is often limited to small case series and case reports.

ASSOCIATION BETWEEN POLYMORPHISM OF THE DNA REPAIR SMUG1 AND UNG GENES AND AGE-RELATED MACULAR DEGENERATION

To investigate the association between the g.4235T>C (rs2337395) polymorphism of the UNG gene and the c.-31A>G (rs3087404) polymorphism of the SMUG1 gene and the risk of age-related macular degeneration (AMD), as well as modulation of this association by some environmental and lifestyle factors. Overall, 272 AMD patients and 105 control subjects were enrolled in this study. Both polymorphisms were genotyped by restriction fragment length polymorphism-polymerase chain reaction (PCR-RFLP). The C/C genotype of the g.4235T>C polymorphism of the UNG gene was associated with an increased risk of dry AMD (odds ratio, 2.54), whereas the T/T genotype of this polymorphism decreased such risk (odds ratio, 0.41). The presence of the T allele of the g.4235T>C polymorphism and the A allele of the c.-31A>G polymorphism of the SMUG1 gene (odds ratio, 2.17 and 2.18, respectively) was associated with an increased risk of AMD severity, expressed by the comparison of the frequencies of genotypes in the group of patients with wet AMD versus those with dry AMD. Conversely, the C/C genotype of the g.4235T>C polymorphism, the G/G genotype of the c.-31A>G polymorphism, and the C/C-G/G combined genotype of both polymorphisms had a protective effect (odds ratio, 0.48, 0.46, and 0.18; respectively). The results obtained suggest the potential role of the g.4235T>C and the c.-31A>G polymorphisms in AMD pathogenesis.

Does CYP1A1 gene polymorphism affect cell damage biomarkers and ageing?

This study aimed to investigate the effects of cytochrome P450 1A1 (CYP1A1) gene polymorphism on early ageing due to occupational exposure to mutagens. The project was conducted on buccal epithelial cells collected from mechanical-workshop workers and controls. Epithelial cells were obtained from the inner part of the cheeks. Techniques including comet assay, micronuclei test, real-time polymerase chain reaction, and restriction fragment length polymorphism-polymerase chain reaction were used to assess DNA damage biomarkers. The results showed that there was a significant difference in relative telomere length (P = 0.039) between wild and mutated genotypes among all individuals. Furthermore, occupational exposure showed no statistically significant effects on biomarkers of the possible influences of gene polymorphism and different rates of ageing among workers (P > 0.05), except for relative telomere length (P = 0.041) among workers as a whole. It was concluded that the effects of occupational exposure to mutagens on genetic material in buccal cells mediated by CYP1A1 gene polymorphism were not significant. In addition, CYP1A1 gene polymorphism might have a protective effect against premature ageing due to occupational exposure to mutagenic chemicals.

GENE POLYMORPHISM OF TRANSFORMING GROWTH FACTOR BETA-1 (TGF-Β1) AND ITS LEVEL IN RHEUMATOID ARTHRITIS PATIENTS ASSOCIATED WITH OSTEOPOROSIS DISEASE

Objective: The imbalance between pro-inflammatory and anti-inflammatory cytokines is a feature of rheumatoid arthritis (RA). The role of transforming growth factor beta-1 (TGF- β1) and its receptor in the pathogenesis of RA is conflicting. We aim to investigate the role of polymorphisms in the gene in susceptibility and severity of RA and osteoporosis.Methods: Eighty-six RA patients and eighty-four control were enrolled in the study.Genotyping of TGF- β1 T/C869 and TGF- β1 G/A800 was determined by restriction fragment length polymorphism-polymerase chain reaction (PCR-RFLP).Results: Subjects carrying T allele of TGF- β1 T/C869 genotype were significantly more likely to have osteoporosis than subjects carrying C allele (OR:6.95, 95% CI: 2.7-15.7, P: 0.001).Also, Subjects with A allele of TGF- β1 G/A800 genotype were significantly more likely to have osteoporosis (OR: 1.13, 95% CI: 1.05-1.2, P: 0.001).Conclusion: T/C869 genotype and G/A800 genotype of TGF-β1 gene affecting on severity of RA and have the ability to develop the risk to osteoporosis in patients carrying T allele and A allele of T/C869 and G/A800 gene polymorphism respectively

Tumour necrosis factor-alpha gene polymorphisms in Iranian patients with biliary atresia.

Biliary atresia (BA) is a progressive inflammatory destructive process of the bile ducts. This study evaluated the relationship between single-nucleotide polymorphisms in the promoter region of tumour necrosis factor-alpha (TNF-α) gene and bilaiary atresia. Genomic deoxyribonucleic acid from 16 patients with established diagnosis of BA and 36 patients with INC was obtained. The genotypes of TNF-α-1031 (T/C) and TNF-α-308 (G/A) were determined using the restriction fragment length polymorphism-polymerase chain reaction and the results were analysis with proper statistic software. The frequencies of T/T, T/C in TNF-α-1031 and G/G, G/A in TNF-α-308 were as same as control group. Moreover, we have same deduction for allele frequency and haplotypes analysis (T allele: 84.37%; G allele: 87.5%) in BA patients (T allele: 80.56%; G allele: 86.11%) in controls. In all cases variants of polymorphism did not affect the severity or incidence of BA disease. although no significant associations were found between BA and control groups, it seems meaningful that since the nature of BA is multi factorial. Next step will be considering a new target such as downstream modulation of the TNF-α pathway or other cytokines and chemokines which act directly/indirectly.

P53 Codon 72 Genetic Polymorphism in Asthmatic Children: Evidence of Interaction With Acid Phosphatase Locus 1.

Several lines of evidence are implicating an increased persistence of apoptotic cells in patients with asthma. This is largely due to a combination of inhibition, or defects in the apoptotic process and/or impaired apoptotic cell removal mechanisms. Among apoptosis-inducing genes, an important role is played by p53. In the present study, we have investigated the possible relationship between p53 codon 72 polymorphism and asthma and the interaction with ACP1, a genetic polymorphism involved in the susceptibility to allergic asthma. We studied 125 asthmatic children and 123 healthy subjects from the Caucasian population of Central Italy. p53 codon 72 and ACP1 polymorphisms were evaluated using a restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method. There is a statistically significant association between p53 codon 72 polymorphism and allergic asthma: Arg/Arg genotype is more represented in asthmatic patients than in controls (P=0.018). This association, however, is present in subjects with low ACP1 activity A/A and A/B only (P=0.023). The proportion of children with A/A and A/B genotype carrying Arg/Arg genotype is significantly high in asthmatic children than in controls (OR=1.941; 95% C.I. 1.042-3.628). Our finding could have important clinical implications since the subjects with A/A and A/B genotypes of ACP1 carrying Arg/Arg genotype are more susceptible to allergic asthma than Pro/Pro genotype.