Restriction Fragment Length Research Articles

Diagnostic utility of exome sequencing for inherited peripheral neuropathies

Introduction. Hereditary motor and sensory neuropathies, a highly genetic heterogeneous group of disorders, have a phenotype caused by peripheral nerve damage.Purpose of the study – to assess the extent of genetic heterogeneity of hereditary motor and sensory neuropathies in Russian patients and to evaluate the diagnostic effectiveness of using full-exome research methods to find the genetic cause of hereditary motor and sensory neuropathies.Materials and methods. The material for the study was DNA samples from 51 patients and their family members referred for whole exome sequencing to the DNA-diagnostics laboratory of Research Centre for Medical Genetics in 2017–2019. Methods: whole exome sequencing, Sanger sequencing, restriction fragment length polymorphism.Results. Whole exome sequencing in combination with segregation analysis of the pathogenic variants in families allowed to determine the cause of the disease in 41 % of cases. In another 16 % of cases, candidate genetic variants as a possible cause of the disease were revealed, but additional studies are needed to confirm it. The most frequently mutated gene was MFN2 caused neuropathy in 6 unrelated families. MPZ gene mutations were detected in two families, AARS gene mutations were revealed in another two families, and mutations in GJB1, HINT1, INF2, LRSAM1, LITAF, MME, NEFL, WWOX were detected once. Among the causal variants, mutations in B4GALNT1 caused spastic paraplegia, in COL6A1 led to Bethlem’s congenital muscular dystrophy, and in SYT2 caused congenital myasthenic syndrome indicating difficulties in differential diagnosis of inherited neuromuscular disorders. A PMP22 duplication was detected in 2 families prior to whole exome sequencing.Conclusion. Whole exome sequencing is very important for finding the molecular cause of hereditary motor and sensory neuropathies. In most cases, additional methods should be used to clarify the pathogenicity of variants detected by whole exome sequencing. However, it is necessary to remember that the most common cause of the disease is a large duplication of the region 17p11.2.

The Inter-Relation between Leptin Receptor (Q223R) Gene Polymorphism and the Risk of Egyptian Patients with HCC.

Background:The relationship of leptin (LEP) and polymorphism of leptin receptor (LEPR) were studied in patients with hepatocellular carcinoma (HCC) and compared with those with liver cirrhosis to find out the extent of the risk of LEPR on patients with HCC. Methods:Serum LEP level and LEPR Q223R gene polymorphism were determined in 300 patients with liver disease categorized equally into five groups’ healthy volunteers, patients with hepatitis C (HCV), patients with non-alcoholic steatohepatitis (NASH), liver cirrhosis and HCC. LEPR gene was amplified by polymerase chain reaction (PCR) then digested by the MSP1 restriction enzyme. Results:The isolated 212 bp of LEPR was sequenced. The serum LEP level was reduced in patients with cirrhotic and HCC. Serum LEP level had negatively correlated with both tumor grade and size in HCC patients. The data obtained from restriction fragment length polymorphismPCR and sequencing revealed the existence of a novel synonymous Q223R single nucleotide polymorphism (SNP) in exon 223 of LEPR gene (1137101). LEPR Gln223Arg, GG and GA genotypes were found in all studied groups. LEPR Gln223Arg, AA genotype was found in NASH, HCC, and control. LEPR Gln223Arg GA genotype is associated with some patients with HCC. Conclusion:GA genotype of LEPR Gln223Arg may be regarded as a probable genetic risk factor for Egyptian patients with HCC.

Association between genetic polymorphisms of telomere pathway genes and hydrogen peroxide level in omethoate exposure workers

ObjectiveThe aim of this study was to explore the association between genetic variations in telomere pathway genes and the level of hydrogen peroxide (H2O2) in omethoate exposure workers. MethodsA total of 180 omethoate exposure workers and 115 healthy controls were recruited. The level of H2O2 in plasma was determined with molybdenic acid colorimetry. Polymerase chain reaction and restriction fragment length was used to detect polymorphisms in POT1 rs1034794, POT1 rs10250202, TERF1 rs3863242, and TERT rs2736098. ResultsThe level of H2O2 in exposure group (4.26 ± 0.71) was significantly higher than that in control group (3.29 ± 0.46). Generalized linear models indicated that risk factors for the increase H2O2 level were exposure [β(95 % CI) = 0.951 (0.806, 1.096), P < 0.001] and AA + AT genotype in POT1 rs034794 [β(95 % CI) = 0.397 (0.049, 0.745), P = 0.025]. ConclusionThe increase H2O2 level was associated with omethoate exposure and AA + AT genotypes in POT1 gene rs1034794. It provided a new idea that polymorphisms in telomere pathway genes may indirectly regulate telomere length by influencing oxidative stress.

Stable high frequencies of sulfadoxine\u2013pyrimethamine resistance associated mutations and absence of K13 mutations in Plasmodium falciparum 3 and 4\xa0years after the introduction of artesunate plus sulfadoxine\u2013pyrimethamine in Ujjain, Madhya Pradesh, India

BackgroundArtesunate plus sulfadoxine–pyrimethamine (ASP) is first-line treatment for uncomplicated Plasmodium falciparum malaria in most of India, except for six North-eastern provinces where treatment failure rates were high. In Ujjain, central India, the frequency of mutations associated with increased drug tolerance, but not overt resistance to sulfadoxine and pyrimethamine were 9% and > 80%, respectively, in 2009 and 2010, just prior to the introduction of ASP. The frequency of drug resistance associated mutations in Ujjain in 2015–2016 after 3–4 years of ASP use, are reported.MethodsBlood samples from patients with P. falciparum mono-infection verified by microscopy were collected on filter-paper at all nine major pathology laboratories in Ujjain city. Codons pfdhfr 16–185, pfdhps 436–632 and K13 407–689 were identified by sequencing. Pfcrt K76T and pfmdr1 N86Y were identified by restriction fragment length polymorphism.ResultsSulfadoxine–pyrimethamine resistance-associated pfdhfr 108 N and 59R alleles were found in 100/104 (96%) and 87/91 (96%) samples, respectively. Pfdhps 437G was found in 10/105 (10%) samples. Double mutant pfdhfr 59R + 108 N were found in 75/81 (93%) samples. Triple mutant pfdhfr 59R + 108 N and pfdhps 437G were found in 6/78 (8%) samples. Chloroquine-resistance-associated pfcrt 76T was found in 102/102 (100%). Pfmdr1 N86 and 86Y were identified in 83/115 (72%) and 32/115 (28%) samples, respectively.ConclusionThe frequency of P. falciparum with reduced susceptibility to sulfadoxine–pyrimethamine remained high, but did not appear to have increased significantly since the introduction of ASP. No polymorphisms in K13 associated with decreased artemisinin susceptibility were found. ASP probably remained effective, supporting continued ASP use.

Vitamin D receptor polymorphisms in spontaneous preterm birth: a case-control study

AimTo evaluate the association between the FokI (rs2228570), ApaI (rs7975232), Bsml (rs1544410), TaqI (rs 731236), and Cdx2 (rs11568820) single nucleotide polymorphisms (SNPs) in the vitamin D receptor (VDR) gene and spontaneous preterm birth (SPTB), as well as their effect on clinical characteristics of women with SPTB and their newborns.MethodsThis case-control study enrolled women who gave birth at the Department of Obstetrics and Gynecology, University Medical Center Ljubljana between 2010 to 2019. Cases were 118 women with spontaneous initiation of PTB after natural conception and 119 controls with a term singleton delivery after an uncomplicated pregnancy. The molecular analysis of VDR SNPs employed polymerase chain reaction and restriction fragment length polymorphism.ResultsPatients and controls did not significantly differ in the distribution of genotype or allele SNP frequencies. However, the FokI polymorphism had a significant effect on newborn birth weight in women with SPTB but not in controls (F = 5.17, P = 0.007, one-way ANOVA with post-hoc Scheffe test), with newborns of FokI TT carriers having the lowest birth weight (P = 0.011). No other VDR SNP was associated with any other clinical characteristic of women with SPTB and their newborns.ConclusionThe TT genotype of the VDR FokI polymorphism is associated with newborn birth weight in women of European origin with SPTB.

GENETIC VARIABILITY OF Pun1 GENE (CAPSAICIN SYNTHASE) IN PUNGENT CULTIVARS OF Capsicum annuum OF NORTHERN MEXICO.

&lt;p&gt;&lt;strong&gt;Background:&lt;/strong&gt; Capsaicinoids are exclusive of the genus &lt;em&gt;Capsicum&lt;/em&gt; and are responsible for fruit pungency. The genetic basis of pungency is not yet fully understood, although several candidate genes have been analyzed including the Pungent gene (&lt;em&gt;Pun1&lt;/em&gt;). &lt;strong&gt;Objective: &lt;/strong&gt;The main aim of this study was to identify the &lt;em&gt;Pun1&lt;/em&gt; in some pungent samples of &lt;em&gt;Capsicum annuum&lt;/em&gt; of Northern Mexico adapted to conditions of high temperature and analyze the variability of polymorphisms in the gene. &lt;strong&gt;Methodology:&lt;/strong&gt; The coding sequence of the &lt;em&gt;Pun1&lt;/em&gt; gene was analyzed by Polymerase Chain Reaction and Restriction Fragment Length. &lt;strong&gt;Results: &lt;/strong&gt;The &lt;em&gt;Pun1&lt;/em&gt; gene showed highly variable patterns; the variable regions were sequenced, and we found 19 single nucleotide polymorphisms in the first exon and 7 in the intron, some polymorphisms generated amino acid changes; while that of Chiltepin cultivar showed more than one sequence. &lt;strong&gt;Implications:&lt;/strong&gt; The protein homology model suggests that these polymorphisms could affect their functionality. &lt;strong&gt;Conclusion:&lt;/strong&gt; All the pungent cultivars of Mexican &lt;em&gt;C. annuum&lt;/em&gt; analyzed were positive to &lt;em&gt;Pun1&lt;/em&gt; gene and have a high variety of polymorphisms, this study provides evidence of the high variability that exists in the &lt;em&gt;Pun1&lt;/em&gt; gene responsible for pungency and could allow the selection of pungent variants to obtain a greater quantity of capsaicinoids for different applications.&lt;/p&gt;

ASSOCIATION BETWEEN MATRIX METALLOPROTEINASE-1 −1607 1G/2G POLYMORPHISM AND CHRONIC PERIODONTITIS IN INDONESIAN POPULATION

Objective: This study aimed to identify the distribution of matrix metalloproteinase (MMP)-1 −1607 1G/2G alleles and genotypes in subjects withchronic periodontitis and healthy subjects in a sample of Indonesian population and assess the possible association of this polymorphism withsusceptibility to chronic periodontitis.Methods: Genomic DNA samples were obtained from 200 Indonesian males aged 33–78 years old, comprising 100 chronic periodontitis patientsand 100 healthy controls. DNA fragments were amplified by a polymerase chain reaction and analyzed by restriction fragment length polymorphism.Results were analyzed by Chi-square test.Results: The frequency of the 2G allele was high both in subjects with periodontitis (87%) and in controls (91%). Analysis of MMP-1 genotype(−1607 1G/2G) showed no significant difference between the chronic periodontitis and healthy groups (p&gt;0.05).Conclusion: The result found no association between MMP-1 −1607 1G/2G polymorphism and susceptibility to chronic periodontitis in Indonesiansubjects.

Evaluation of SCD, ACACA and FASN Mutations: Effects on Pork Quality and Other Production Traits in Pigs Selected Based on RNA-Seq Results.

Simple SummaryThis study aimed to evaluate mutations within three candidate genes (SCD, ACACA, FASN) for their effects on fattening and slaughter characteristics, as well as meat quality traits, including intramuscular fat (IMF) level in pork. They were selected within differentially expressed genes activated in response to variable backfat content obtained using the RNA sequencing method. The RNA-seq analysis identifies mutations/SNPs located in the mRNA and could be a useful tool for prediction of genetic markers in farm animals. The results showed that selection for FASN A allele in Polish Large White pigs could lead to improved meat quality traits such as water exudation and meat colour. However, analysed polymorphisms showed only slight effects on fat metabolism and IMF content.In recent years, pig producers have struggled with the problem of low intramuscular fat levels in pork, which impacts palatability and ultimately meat quality. Reduced levels of intramuscular fat are likely the result of breeding objectives aimed at increasing lean meat content. In this study, three mutations within candidate genes for fat content (SCD, ACACA, and FASN) were selected, based on RNA-seq results and the relationship between polymorphisms in genes related to lipid metabolism, fattening and slaughter characteristics, as well as pork quality, including IMF level, were evaluated to identify selection markers. Moreover, their impact on gene expression was also examined. The PCR–RFLP (polymerase cha- in reaction – restriction fragments length) method was used to establish genotypes and effect sizes of potential genetic markers were estimated using a GLM model. It was identified that a FASN missense variant was positively associated with the expression level of this gene, which suggested its linkage with a mutation having a regulatory function. The association study indicated that the FASN missense variant may play a role in the determination of feed conversion and meat colour. In turn, a mutation in the ACACA gene showed a relationship with IMF content in the Puławska breed where the differences reached as much as 20%. We suggest considering all three mutations in further studies based on different pig populations due to the crucial role of SCD, ACACA, and FASN genes in lipid metabolism.

Association of the I/D polymorphism of angiotensinconverting enzyme gene with the development of essential hypertension

Objective. To determine the impact of the I/D polymorphism of the angiotensin-converting enzyme (ACE) gene on the development of essential hypertension, taking into account gender differences.Material and Methods. Clinical data were assessed and a molecular genetic study was performed in 602 people including 401 patients with essential hypertension and 201 individuals of the control group, representing the Belarusian ethnic group. Genotyping was performed using the method of polymerase chain reaction and restriction fragment length polymorphism.Results. The distribution of genotypes of the I/D polymorphism of the ACE gene did not differ between patients with hypertension and normotensive individuals: II, ID, and DD genotypes were detected in 100 (24.9%), 192 (47.9%), and 109 (27.2%) patients and in 52 (25.9%), 108 (53.7%), and 41 (20.4%) people of the comparison group, respectively. Differences were found between the distribution of DD genotype in men with hypertension and in the control group, where the frequencies were 28.4% and 17.3% (p = 0.04), respectively, in contrast to no differences in women: 25.8% and 23.3% (p = 0.64), respectively. Carrying the DD genotype in men compared with the ID and DD genotypes (recessive model) of the I/D polymorphism of the ACE gene increased the probability of developing essential hypertension by 1.9 times (OR = 1.89; 95% CI = 1.04-3.44). The analysis of the prevalence of risk factors depending on the I/D polymorphism of the ACE gene showed that male patients with the DD genotype more often had burdened heredity in regard to the development of premature cardiovascular diseases (23 patients (37.7%)) compared with the individuals with II and ID genotypes: 13 (21.7%) and 14 (14.9%) patients, respectively (χ2 = 1.16; p = 0.005), and mainly through the paternal line.Conclusions. Development of essential hypertension is associated with the carriership of the mutant DD genotype of I/D polymorphism of the ACE gene in men.

Abstract P017: Biomarkers Representing Aging-Related Biological Pathways Are Associated With All-Cause Mortality: The Framingham Study

Background: Chronological aging is the result of different biological processes and molecular mechanisms that, if identified, may increase our ability to mitigate aging-related decline in body functions. We hypothesized that aging-related biomarkers reflecting multiple biological pathways are associated with all-cause mortality. Methods and Results: We related leukocyte telomere length (LTL) expressed by terminal restriction fragment (TRF) length, urinary concentrations of creatinine-corrected isoprostanes, circulating concentrations of insulin-like growth factor (IGF)-1, and asymmetrical dimethylarginine (ADMA) in up to 2314 Framingham Offspring Study participants attending routine examination cycles (mean age 58 yrs, 55% women) to all-cause mortality using Cox proportional hazards regression models. Biomarkers were modeled individually and in combination, adjusting for age, sex, body mass index, systolic blood pressure, diabetes, smoking status, hypertension medication, total cholesterol/HDL. We created also a biomarker score as a composite of biomarkers associated with mortality risk. Kaplan Meier curves were created ( Figure ) to graphically present the survival time as a function of tertiles of the biomarker score. There were 593 deaths (274 women) during a median follow-up of 20 years. TRF and IGF-1 values were inversely related to all-cause mortality (multivariable-adjusted hazard ratios [HR] per SD increase, 0.86, 95% confidence interval [CI], 0.75-0.998 and 0.89, 95% CI 0.81-0.97 for TRF and IGF-1, respectively). Isoprostanes and ADMA values were positively related to all-cause mortality (multivariable-adjusted HR per SD increase, 1.17, 95% CI, 1.11-1.23, and 1.09, 95% CI, 1.01-1.18, respectively). Conclusion: In our prospective community-based study, aging-related biomarkers were associated with all-cause mortality, supporting the concept that molecular pathways represented by these biomarkers may reflect the processing of aging in the community.

&lt;p&gt;MTHFR C677T polymorphism and risk of nonsyndromic cleft lip with or without cleft palate in the Moroccan population&lt;/p&gt;

BackgroundNonsyndromic cleft lip with or without cleft palate (NSCL/P) is one of the most common craniofacial malformations observed. Several studies suggest that the decrease in folate has been associated with a higher risk of NSCL/P. The present study aimed to determine the association of 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism gene with the occurrence of NSCL/P in the Moroccan population.MethodsMTHFR C677T was genotyped in 52 Moroccan patients and 182 unrelated controls, using a PCR followed by restriction fragment length polymorphism.ResultsThe results of the study revealed a genotypic and phenotypic distribution in equilibrium with Hardy–Weinberg’s law (χ2=0.36, P=0.55). The frequency of heterozygous genotype C/T and the T allele in controls and patients were 40.7% vs 15.4% and 26%, respectively.ConclusionA low association was found between the C677T polymorphism of the MTHFR gene and a risk for the development of NSCL/P in the Moroccan population (OR =0.24, P=0.0005).

IMPLICATION OF FIVE AIDS RELATED GENES IN MOTHER-TO-CHILD TRANSMISSION AND ACQUISITION OF HUMAN IMMUNODEFICIENCY VIRUS 1 IN CAMEROON.

Background:Genetic variants in the mother and/or infant have been described with evidence to be associated with mother-to-child transmission of HIV, but somehow with contradictory results depending on ethnic or geographic populations. We aimed at looking at the association between the allelic frequency of some genes with vertical transmission or acquisition of HIV in Cameroon.Methodology:A total of 262 mothers (212 HIV-infected and 50 HIV non-infected) with their babies (270 in total, 42 HIV exposed-infected, 178 HIV exposed non-infected and 50 HIV non-exposed) were recruited in Yaounde-Cameroon. Their genotypes for CCR5-Delta32, CCR5 promoter59029A/G, CCR2-64I, SDF1-3’A and TRIM5α-136Q were analyzed using polymerase chain reaction and restriction fragment length polymorphisms.Results:Allelic frequencies were 14.7%, 41.9%, 9.5% and 14.7% for CCR2-64I, CCR5-59029-A/G, TRIM5α-136Q, SDF1-3’A respectively in the mothers and 18.8%, 35.9%, 11.3% and 20.5% in the babies. No delta 32 mutation in the CCR5 gene was found. The mutant genotype was most significantly frequent in the non-transmitter than in the transmitter (p= 0.005) for the SDF-1 3’A. SDF1-3’A [Odd ratio = 1.69; 95% confidence interval: 0.1158 to 0.7277); was associated to MTCT, P = 0.008.The homozygote mutants for the CCR5-59029-G were significantly higher in the infected than in the exposed uninfected babies (p=0.04). The mutations in the other genes were neither implicated in the acquisition nor in the transmission.Conclusion:SDF1-3’A was associated to the reduction of MTCT. The CCR5-59029-A/G favored acquisition of HIV by babies. Our study showed that polymorphisms in chemokine ligand may be involved in MTCT.

Serum 25-hydroxyvitamin D, serum calcium and vitamin D receptor (VDR) polymorphisms in a selected population with lumbar disc herniation-A case control study.

BackgroundAssociation of Vitamin D receptor (VDR) polymorphisms with lumbar disc herniation (LDH) have been identified in several ethnic groups globally. Despite abundant sunlight, vitamin D deficiency is reported in many tropical countries. As vitamin D is a key modulator for intestinal calcium absorption, low vitamin D could contribute to low serum calcium leading to abnormalities of skeletal homeostasis. Therefore, present study was aimed to study the association of serum 25-hydroxyvitamin D (25(OH)D), serum calcium and VDR polymorphisms in a selected Sri Lankan population.Materials & methodsA case control study was conducted in 119 participants (cases = 51: controls = 68). Serum 25(OH)D levels were measured using ELISA. The VDR polymorphisms (Fok I and Taq I) were detected by polymerase chain reaction followed by restriction fragment length polymorphism.ResultsFindings indicated a significantly low (p = 0.000) 25(OH)D levels in cases (18.7±3.7 ng/mL) compared to controls(25.5±9.8 ng/mL) while 25(OH)D in both groups were below the reference range. Mean serum calcium levels in both groups were within normal reference range and was not significantly different among groups. Statistically significant association was not observed between VDR Fok I polymorphisms among cases and controls. Although Fok I polymorphism genotypes were in Hardy-Weinberg equilibrium (HWE), Taq I genotypes in controls violated HWE.ConclusionPresent study confirms that insufficient serum 25(OH)D levels in cases have major contribution to LDH. VDR Fok I polymorphisms did not have any significant association with LDH in Sri Lankan ethnicity.

Polymorphism of the adiponutrin gene (PNPLA3) in the indigenous inhabitants of the Republic of Sakha (Yakutia) with type 2 diabetes mellitus

Rationale:The association of rs738409 I148M polymorphism with type 2 diabetes mellitus (T2DM) and non-alcoholic fatty liver disease has been confirmed for several ethnic and territorial groups. Up to now, no such studies have been performed in the populations of Yakutia.Aim:To study allele frequency distribution and to identify associations of the PNPLA3 gene polymorphism (rs738409 C&gt;G) with T2DM in the Yakuts.Materials and methods: DNA samples from 106 T2DM patients were used in the study; the control group included samples from 72 healthy volunteers. All study participants were ethnic Yakuts and were living in the territory of the Republic of Sakha (Yakutia), Russian Federation. rs738409 polymorphism of the PNPLA3 gene was studied by polymerase chain reaction and by restriction fragment length polymorphism.Results:There were no significant difference in the distribution of the allele frequencies and genotypes of the polymorphous variant of the PNPLA3 gene (rs738409) between the T2DM patients and the healthy control. Both groups showed prevailing allele G (р = 0.01) and homozygous genotype GG (96%).Conclusion:High frequency of the allele G (74.1%) with predominance of GG genotype (58.5%) was found in type 2 diabetic patients.

COMT Val 108/158 Met polymorphism and treatment response to aripiprazole in patients with acute schizophrenia.

IntroductionThe COMT Val 108/158 Met polymorphism (rs4680) may affect treatment response to antipsychotics, as well as metabolism and dynamics of neurotransmitters during the treatment of schizophrenia. We investigated the effects of the COMT Val 108/158 Met polymorphism on treatment response to aripiprazole and plasma monoamine metabolite levels in patients with acute schizophrenia.Materials and methodsForty patients with schizophrenia were treated with aripiprazole for 6 weeks. We measured Positive and Negative Syndrome Scale (PANSS) and plasma levels of homovanillic acid (HVA) and plasma MHPG (3-methoxy-4-hydroxyphenethyleneglycol) at baseline and endpoint. The COMT Val 108/158 Met polymorphism was genotyped with the polymerase chain reaction and restriction fragment length polymorphism.ResultsThere were significant genotype–time interactions on PANSS total and general psychopathology scores, with Met/Met genotype showing greater improvement. The response rate to aripiprazole did not differ between COMT Val 108/158 Met genotype groups. We found a significant time effect on plasma MHPG levels, but no time effect on plasma HVA levels or time–genotype interactions in the plasma levels of HVA and MHPG. Although the responder rate did not differ among the 3 genotype groups.ConclusionOur results suggest that individuals with the Met/Met genotype had greater improvement in PANSS score after the treatment with aripiprazole. On the other hand, the Val 108/158 Met polymorphism may not induce changes in plasma levels of monoamine metabolites during aripiprazole treatment. Because of the small sample size, further studies are needed to confirm and to extend our results.

The role of COX-2 gene variants on the disease mechanism of inflammatory bowel disease in a Turkish population

Aim: Inflammatory bowel disease has two major types: Crohn’s disease and ulcerative colitis that occur in the gastrointestinal tract with unknown etiology. COX-2 has important role on carcinogenesis process including colon cancer supporting the tumor growth. COX-2 was also known due to its ability to change homeostasis on colonic mucosa in inflammatory cells on patients who have inflammatory bowel disease. In this study, we have aimed to find a linkage between inflammatory bowel disease and COX-2 in a Turkish population. Methods:A total of 106 patients,42 with Crohn’s disease and 64 with ulcerative colitis and 121 healthy control subjects were included the study. Gene variants of COX-2-765G→C and COX-2-1195A→G were analyzed by polymerase chain reaction and restriction fragment length polymorphism techniques.Results: The results demonstrated that COX-2-1195A→G gene variants AA carriers were statistically found in high level on patients with both ulcerative colitis (p=0,001) and Crohn’s disease (p=0.008). In contrast, AG genotype and G carriers were statistically found higher in control group (Crohn’s disease, p=0.005 for AG and p= 0.008 for G; ulcerative colitis, p=0.001 for AG and p=0.001 for G). Conclusion: In this research, we have observed important and questionable results between inflammatory bowel disease and COX-2, especially COX-2-1195A→G gene variants AA carriers in a Turkish population. Researches need to focus on their local roles on inflammatory bowel disease pathogenesis with large sample size.

Telomere length is independently associated with age, oxidative biomarkers, and sport training in skeletal muscle of healthy adult males

In skeletal muscle, which mainly contains postmitotic myonuclei, it has been suggested that telomere length remains roughly constant throughout adult life, or shortens in response to physiopathological conditions in muscle diseases or in the elderly. However, telomere length results from both the replicative history of a specific tissue and the exposure to environmental, DNA damage-related factors, therefore the predictive biological significance of telomere measures should combine the analysis of the various interactive factors. In the present study, we analysed any relationship between telomere length [mean and minimum terminal restriction fragment (TRF) length] chronological age, oxidative damage (4-HNE, protein carbonyls), catalase activity, and heat shock proteins expression (αB-crystallin, Hsp27, Hsp90) in semitendinous muscle biopsies of 26 healthy adult males between 20 and 50 years of age, also exploring the influence of regular exercise participation. The multiple linear regression analysis identified age, 4-HNE, catalase, and training status as significant independent variables associated with telomere length and jointly accounting for ∼30–36% of interindividual variation in mean and/or minimum TRF length. No association has been identified between telomere length and protein carbonyl, αB-crystallin, Hsp27, and Hsp90, as well as between age and the variables related to stress response. Our results showed that skeletal muscle from healthy adults displays an age-dependent telomere attrition and that oxidised environment plays an age-independent contribution, partially influenced by exercise training.

Association between polymorphism of tumor necrosis factor alpha (tnfα) in the region -308 g/a with tinnitus in the elderly with a history of occupational noise exposure.

Context:Tinnitus is a common disorder that occurs frequently across all strata of population and has an important health concern and is often associated with different forms of the hearing loss of varying severity.Aims:To investigate the association between the polymorphism of tumor necrosis factor alpha (TNFα) in the region −308 G/A with the susceptibility to tinnitus in individuals with the history of exposure to occupational noise.Settings and Design:This was a cross-sectional study with a sample of 179 independent elderly people above 60 years of age.Materials and Methods:Information on exposure to occupational noise was obtained by interviews. Audiological evaluation was performed using pure tone audiometry and genotyped through polymerase chain reaction by restriction fragment length polymorphism.Statistical Analysis Used:Data were analyzed using the chi-square test and the odds ratio (OR), with the significance level set at 5%.Results:Among elderly with tinnitus (43.01%), 33.76% had a history of exposure to occupational noise. A statistically significant association was found between genotype frequencies of the TNFα gene in the −308 G/A region and the complaint of tinnitus (P = 0.04 and χ2 = 4.19). The elderly with the G allele were less likely to have tinnitus due to occupational noise exposure when compared to those carrying the A allele (OR = 2.74; 95% CI: 1.56–4.81; P < 0.0005).Conclusion:This study suggests an association between the TNFα with susceptibility to tinnitus in individuals with a history of exposure to occupational noise.

Long-term dynamics of the bacterial community in a Swedish full-scale wastewater treatment plant

ABSTRACTThe operational efficiency of activated sludge wastewater treatment plants depends to a large extent on the microbial community structure of the activated sludge. The aims of this paper are to describe the composition of the bacterial community in a Swedish full-scale activated sludge wastewater treatment plant, to describe the dynamics of the community and to elucidate possible causes for bacterial community composition changes. The bacterial community composition in the activated sludge was described using 16S rRNA gene libraries and monitored for 15 months by a terminal restriction fragment (T-RF) length polymorphism (T-RFLP) analysis of the 16S rRNA gene. Despite variable environmental conditions, a large fraction of the observed T-RFs were present at all times, making up at least 50% in all samples, possibly representing a relatively stable core fraction of the bacterial community. However, the proportions of the different T-RFs in this fraction as well as the T-RFs in the more variable fraction showed a significant variation over time and temperature. The difference in community composition between summer and winter coincided with observed differences in floc structure. These observations suggest a relationship between floc properties and bacterial community composition, although additional experiments are required to determine causality.

Association of the gene polymorphism of cytotoxic T lymphocyte-associated antigen-4with Graves ophthalmopathy in Qinghai Tibetans

To investigate the association of the gene polymorphism of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4)with Graves′ ophthalmopathy(GO)in Qinghai Tibetans. 130 case of Graves′ disease(GD)were selected from June 2013 to November 2016 in The People′s Hospital of Qinghai Province. These patients were assigned into 2 groups, GO(n=71)and GD without GO(n=59). The genotypes and alleles of CTLA-4 gene were detected by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Distribution of CTLA-4 exon 1(+ 49A/G)genotype frequencies(AA, AG, and GG)was different between 2 groups. (Chin J Endocrinol Metab, 2017, 33: 962-964) Key words: Graves′ disease; Polymorphism, restriction fragment length