TNF-alpha Gene Research Articles

Expression and functional characterization of vitronectin gene from Japanese flounder (Paralichthys olivaceus)

Vitronectin (Vtn) is a multifunctional protein that plays significant roles in cell adhesion, migration, spreading and survival, and in the regulation of membrane attack complex formation and the terminal pathway of complement activation in innate immune response. However, the expression and immune significance of Vtn in fish remains largely unknown. In order to understand the involvement of Vtn in fish innate immune response, here we cloned and characterized a full-length Vtn ortholog cDNA, termed PoVtn, from Japanese flounder Paralichthys olivaceus. The deduced PoVtn protein is comprised of 438 amino acids with a 19-amino-acid signal peptide sequence (1Met-19Ala) at the N-terminus. Protein domain analysis revealed that PoVtn possesses a conserved N-terminal somatomedin B domain followed by a conserved RGD motif and four haemopexin-like domains. Sequence analysis revealed that PoVtn has two potential glycosylation sites and shares 44–74% sequence identity with other teleost Vtn proteins. PoVtn mRNA was ubiquitously distributed in all examined normal tissues and showed the highest expression in Japanese flounder hepatopancreas tissue. PoVtn expression was induced by LPS and poly(I:C) challenges in the Japanese flounder head kidney macrophages (HKMs) and peripheral blood leucocytes (PBLs) and shows a pathogen-associated molecular pattern- and cell type-dependent manner. The expression of PoVtn was also modulated by bacterial challenge with Edwardsiella tarda in Japanese flounder immune-related tissues including head kidney, gill and spleen. Furthermore, overexpression of PoVtn in Japanese flounder FG-9307 cells significantly attenuated the LPS- and poly(I:C)-induced proinflammatory cytokines IL-1beta and TNF-alpha gene expression. Taken our findings together, we for the first time characterized Vtn gene expression in response to inflammatory stimuli in fish. Our results suggested a potential role of PoVtn in regulating fish innate immunity.

Evaluation of TNF-alpha gene (G308A) and MBL2 gene codon 54 polymorphisms in Turkish patients with tuberculosis.

MBL acts as a binding protein that enables uptake of mycobacteria into macrophages. And, TNF-alpha is an important cytokine that is involved in control of mycobacterial infections both in-vivo and in-vitro. A large number of genetic factors exerting susceptibility to tuberculosis has been identified, among which mannose-binding lectin and tumor necrosis factor-alpha call attention. The objective of this study is to compare the frequency of TNF-alpha and MBL gene polymorphisms between patients diagnosed with tuberculosis and healthy volunteers in Turkey, and determine the association between tuberculosis and TNF-alpha gene (G308A) and MBL2 gene codon 54 polymorphisms. The study included 69 patients who were diagnosed with tuberculosis and 70 control subjects. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect TNF-alpha (G308A) gene and MBL2 gene codon 54 polymorphisms. For statistical analysis, the significance level was determined as p<0.05. A comparison between patient and control groups in TNF-alpha (G308A) gene and MBL2 gene codon 54 polymorphisms showed no statistically significant difference (p>0.05). However, a comparison of mean body mass index (BMI) and smoking status showed a statistically significant difference between the tuberculosis and control groups (p=0.01 and p=0.009, respectively). Our results suggest that the MBL2 gene Codon 54 and TNF-alpha gene G308A polymorphisms are not associated with an increased risk for development of tuberculosis in our patients. Further studies are required including more cases of tuberculosis patients and other potentially relevant gene polymorphisms.

The -863 TNF-&amp;alpha; Polymorphism and Primary Open Angle Glaucoma in Egyptians

Growing evidence indicates that TNF-alpha is involved in the pathogenesis of POAG in several ways, primarly by induction of retinal ganglion cells apoptosis and therefore optic nerve degeneration. TNF-alpha and POAG relationship has been studied at the genetic level with variable results in different populations. The transcription rate and the release of the TNF-alpha cytokine have been reported to be affected by polymorphisms in the promoter of the TNF- alpha gene. Polymorphisms at positions -238 and -308 are the most frequent studied. Another polymorphism, at the position -863 in the promoter region, has been less studied, but a homozygous AA allele appears protective in a Chinese population. Our aim was to assess the potential association of -863C/A TNF-alpha gene promoter polymorphisms with POAG in an Egyptian group of subjects. Genotyping of the TNF-alpha (-863) polymorphism was done for 228 POAG patients and 230 control subjects using the PCR-based, Restriction Fragment Length Polymorphism (RFLP) assay. TNF-alpha (-863) A/A genotype was absent in both groups. There was no significant difference between both groups as regards to TNF-α (-863) A allele carriage (6.14 versus 10.43%; p = 0. 099)). Also the genotype TNF-α (-863) C/C and the frequency of the tumor necrosis factor-alpha (-863) C allele did not significantly differ between both groups (93.86 versus 89.57%; p = 0. 099) and 96.93 versus 94.78%; p = 0. 107) respectively. Our data indicated that the TNF-alpha (-863) a allele is not linked with primary open angle glaucoma protection among Egyptian patients.

Lentivirus-mediated short hairpin RNA interference targeting TNF-alpha in macrophages inhibits particle-induced inflammation and osteolysis in vitro and in vivo.

BackgroundAseptic loosening is a significant impediment to joint implant longevity. Prosthetic wear particles are postulated to play a central role in the onset and progression of periprosthetic osteolysis, leading to aseptic loosening of the prosthesis.MethodsWe investigated the inhibitory effects of a lentivirus-mediated short hairpin RNA that targets the TNF-alpha gene on the particle-induced inflammatory and osteolytic changes via macrophages both in vitro and in vivo. An siRNA sequence targeting the mouse TNF-alpha gene from four candidates, transcribed in vitro, was screened and identified. A lentivirus vector expressing short hairpin RNA (shRNA) was then constructed in order to facilitate efficient expression of TNF-alpha-siRNA. Lentivirus-mediated shRNA was transduced into cells of the mouse macrophage line RAW 264.7. Ceramic and titanium particles were introduced 24 h after lentivirus transduction to stimulate cells. TNF-alpha expression, represented by both mRNA and protein levels, was quantified with real-time PCR and ELISA at all time intervals. Lentivirus-mediated shRNA suspension was locally administered into the murine calvarial model, followed by local injection of particles. A multi-slice spiral CT scan was used to evaluate the osteolysis of the calvaria by detecting the width of the cranial sutures.ResultsMacrophages developed pseudopods when co-cultured with particles. Lentivirus-mediated shRNA was shown to effectively inhibit the expression of TNF-alpha at both the mRNA and protein levels in RAW 264.7. The multi-slice spiral CT scan showed that the lentivirus-mediated shRNA significantly suppressed osteolysis of mouse calvaria.ConclusionsOur investigation highlighted the results that lentivirus-mediated shRNA targeting the TNF-alpha gene successfully inhibited particle-induced inflammatory and osteolytic changes both in vitro and in vivo. Therefore, lentivirus-mediated gene therapy may provide a novel therapeutic approach to aseptic joint loosening.

TNF-Alpha Gene Polymorphism in Indian Population during the Course of Pandemic Influenza a H1N1 Infection: A Pilot Study

TNF-Alpha Gene Polymorphism in Indian Population during the Course of Pandemic Influenza a H1N1 Infection: A Pilot Study

Avian recombinant virus H5N1 influenza (A/Vietnam/1203/04) and its escape-mutant m13(13) induce early signaling reactions of the immunity in human lymphocytes

The innate immune receptors TLR4, TLR7, TLR8, and RIG1 recognized the structures of the influenza viruses in human lymphocytes and were activated by the recombinant avian influenza virus A/Vietnam/1203/04 and its escape-mutant m13(13) during early period of interaction. The stimulated levels are not connected with viral reproduction. Donor cells with the low constitutive immune receptors gene expression levels showed higher stimulation. Inflammation virus effects resulted in. increasing production of TNF-alpha and IFN-gamma by lymphocytes. Signaling gene reactions of the parent and mutant viruses endosomal as well as cytoplasmic receptors are very similar. The mutant virus A/Vietnam/1203/04 (HA S145F), stimulated an increase in the transcription level of the membrane receptor gene TLR4 and a decrease in the level of activation of TNF-alpha gene. Further studies of natural influenza virus isolates are necessary to estimate the role of HA antigenic changes on immune reactions in humans.

Polymorphism and expression of the tumour necrosis factor-alpha (TNF-alpha) gene in non-infected cows and in cows naturally infected with the bovine leukaemia virus (BLV)

The objective of this study was to determine the influence of TNF-alpha gene polymorphism at position -824 A > G on TNF-alpha gene expression in BLV-positive animals with aleukaemic (AL) and persistent lymphocytosis (PL) forms of enzootic bovine leukosis (EBL) and in BLV-negative cows. Polymorphism of the TNF-alpha gene at position -824 A > G had a complex influence on TNF-alpha gene expression in cows infected with BLV. In animals with various TNF-alpha genotypes, mRNA levels were stable and similar, but significant differences were noted in the percentages of PBMCs expressing membrane TNF-alpha (mTNF-alpha) protein (PBMCs that were positive for mTNF-alpha). In healthy cows, significant differences in TNF-alpha gene expression were not noted at any of the analysed levels. BLV infection and EBL progression resulted in differential TNF-alpha gene expression at both mRNA and protein levels, but the differences in the amount of the transcript and the percentages of mTNF-alpha+ cells exhibited a reverse trend. The lowest mRNA levels and the highest percentage of PBMCs expressing mTNF-alpha protein were determined in BLV-positive cows. These complex results regarding TNF-alpha gene expression in BLV-positive cows could be attributed to the presence of virus or viral protein/proteins modifying the expression of the TNF-alpha gene.

Association of TNF-alpha and IL-1beta genes polymorphism in Parkinson's disease

Association of TNF-alpha and IL-1beta genes polymorphism in Parkinson's disease

2015 AHA Late-Breaking Basic Science Abstracts.

### 23060 ### Validation Of Circulating MicroRnas As Biomarkers In Heart Failure In Two Large Independent Cohorts Antoni Bayes-Genis1, David Lanfear2, Maurice de Ronde3, Josep Lupon1, Joost Leenders4, Elisabet Zamora1, L. Keoki Williams2, De Antonio Marta5, Zhen Liu4, Koos Zwinderman3, Sara-Joan Pinto-Sietsma3, Yigal Pinto3; 1Universitat Autonoma de Barcelona, Barcelona, Spain; 2Henry Ford Hospital, Detroit, MI; 3Academic Med Cntr, Univ of Amsterdam, Amsterdam, Netherlands; 4Mirnext BV, Amsterdam, Netherlands; 5Germans Trias i Pujol Hosp, Badalona, Spain Background: Small studies suggested circulating microRNAs (circmiRs) as biomarkers for Heart Failure (HF). However, standardized approaches and quality assessment are not established, and results have been inconsistent, with little replication between studies. We aimed to implement quality standards to enable comparison between cohorts and assess which circmiRs may add prognostic information in HF. Methods: We measured 15 circmiRs in two independent cohorts totaling >2000 subjects. Cohort I (Barcelona) comprised of n=843 chronic HFrEF patients. Cohort II from Detroit comprised n= 1384 chronic HF patients (892 HFrEF, 492 HFpEF). Each sample was measured in duplicate, and normalized to an abundant and stable circmiR (miR-486-5p). Algorithms were installed to define each circmiR measurement as “valid”, “unmeasurable” or “invalid”. This allowed inclusion of valid low-level circmiR measurements while reducing noise from false amplification signals. Results: In general, between 20–40% of measurements were “invalid”, while miR-499a\_5p and -208a were “unmeasurable” in the majority of patients in both cohorts. Higher levels of circmiRs-133b, -1254, -622, -208a and -499a\_5p were significantly associated with risk of death in both cohorts, with hazard ratios ranging from 1.103 to 1.365 per log increase (p-values 0.001 to 0.05). However, adding these circmiRs to established predictors (age, renal function and NTproBNP) did not further augment the c-stat beyond 0.71 (cohort I) or 0.78 (cohort II). …

T93: Immune system contributes to the efficacy of cancer chemotherapy

Background Risk of metastasis formation is provided by both tumor cell biological characteristics and the microenvironment features within the primary tumor along with local and systemic conditions for metastatic niche formation. The inflammatory infiltration has been shown to strongly impact on tumor progression (Whiteside, 2013). Dronca et al. (2011) showed that immunosuppressive factors in the tumor microenvironment may impair not only local immune responses but also disturb systemic immunity. Zitvogel et al. anticipate that the comprehension of the mechanisms governing the immunogenicity of cell death will have a profound impact on the design of anticancer therapies.To study the impact of immune system on clinical response to neoadjuvant chemotherapy and metastasis-free survival in breast cancer patients. Materials and methods 350 patients with newly diagnosed invasive breast cancer treated with neoadjuvant chemotherapy (NAC) were enrolled into the study. The procedures were made in accordance with the Helsinki Declaration. Clinical response to chemotherapy, the 5-year metastasis-free survival and all major clinical and morphological parameters were determined. The original method of multidimensional data visualization was applied to present the immune system state as integral entirety in visual image for classification of patients with different risk of metastasis (NovoSpark Corporation, Canada). Copy number aberrations (CNA) of cytokine gene regions in tumor specimens were tested using high-density microarray platform CytoScanTM HD Array (Affymetrix, USA). Cytokine gene polymorphism was analyzed. Subpopulations of lymphocytes and macrophages were determined within the primary tumors by IHC. Results We found, that favorable clinical immediate response to preoperative chemotherapy was related to the high levels of IL-1beta, TNF-alpha and IL-10 production by peripheral mononuclear cells before the treatment. This correlation was further confirmed by data from the study on association between cytokine gene functional polymorphism and response to NAC. We used NovoSpark Corporation visualization approach allowing the representation the immune system state as integral unit and to discriminate breast cancer patients with high and low risk of haematogeneic metastasis. When estimated before cancer treatment, 95% of breast cancer patients had risk of metastasis. The neoadjuvant chemotherapy and surgical tumor removal reduced the risk of tumor progression to 62–71%. However, in a year after adjuvant chemo- and radiotherapy, the patient group with high risk of metastases increased to 81% again. Thus, the cancer treatment can change the primarily estimated outcome prognosis in breast cancer patients, and the monitoring of immune system is a promising approach to predict the risk of cancer progression or resistance to the therapy. We have found the connection between the profile of intra-tumor inflammatory elements and chemotherapy efficacy.Cytokine gene expression may be influenced by the chromosome anomalies (CNA – Copy Number Aberration) – deletion and amplification – of cytokine gene loci in tumor cells. We found the close relation between the clinical response to NAC and gain of function of IL-10 and CHI3L1 (YKL40) genes. In contrast, loss of TNF-alpha and IL-17 gene function due to corresponding CNA was associated with good response to NAC. Metastasis-free survival of breast cancer patients was shown to be closely related to CNA. Conclusion The parameters of the activation of systemic and intra-tumoral immune system by growing tumor and its dissemination have to be validated in order to identify the new prognostic markers for the efficiency of the neoadjuvant chemotherapy.

Association of TNF-alpha and IL-13 genes polymorphisms with bronchial asthma

OBJECTIVE To assess the association of polymorphisms of TNF-alpha gene (rs1799724, rs1800630, rs1799964 and rs769178) and IL-13 gene (rs2158177 and rs1295687) with susceptibility to asthma among ethnic Chinese in Qingdao region. METHODS For 400 asthma patients and 200 healthy subjects, above polymorphisms were detected with a SNaPshot method. RESULTS For rs2158177, the frequency of genotype of GG in the asthma group was significantly lower than the control group (2.8% vs. 5%, OR = 0.31, 95%CI: 0.12-0.82, P = 0.021). No significant difference was detected in the genotypic frequencies for the remaining 5 polymorphisms between the two groups (All P > 0.05). CONCLUSION The study has indicated that rs2158177 polymorphism of the IL-13 gene is associated with asthma in ethnic Han Chinese from Qingdao. No association has been found between polymorphisms of TNF-alpha gene with susceptibility to asthma.

Integrated probability of coronary heart disease subject to the -308 tumor necrosis factor-alpha SNP: a Bayesian meta-analysis.

We present a meta-analysis of independent studies on the potential implication in the occurrence of coronary heart disease (CHD) of the single-nucleotide polymorphism (SNP) at the −308 position of the tumor necrosis factor alpha (TNF-alpha) gene. We use Bayesian analysis to integrate independent data sets and to infer statistically robust measurements of correlation. Bayesian hypothesis testing indicates that there is no preference for the hypothesis that the −308 TNF-alpha SNP is related to the occurrence of CHD, in the Caucasian or in the Asian population, over the null hypothesis. As a measure of correlation, we use the probability of occurrence of CHD conditional on the presence of the SNP, derived as the posterior probability of the Bayesian meta-analysis. The conditional probability indicates that CHD is not more likely to occur when the SNP is present, which suggests that the −308 TNF-alpha SNP is not implicated in the occurrence of CHD.

P42. TNF-ALPHA gene polymorphism in pathology of preeclampsia

Introduction Preeclampsia (PE) is a life-threatening complication of pregnancy associated with a high rate of maternal and perinatal morbidity and/or mortality worldwide. If untreated, it can progress to eclampsia, which can result in the death of mother, fetus or both. Objectives Etiology of PE is still uncertain, however recently the role of immune system takes on importance. Thus the role of central cytokine, tumor necrosis factor (TNF) involved in inflammation processes become widely investigated in obstetric disorders. Following this the present study is to investigate the effect of TNF-alpha gene G308A (rs1800629) polymorphism on disease risk, renal functions, microvascular permeability, endothelial cell dysfunction and organ involvement in women with preeclampsia. Methods Initially a 102 3 rd trimester pregnant women (preeclamptic-cases and healthy controls) with singleton pregnancy were invited for participation of which 76 were genotyped for TNF-alpha G308A polymorphism, evaluated for plasma levels of sVCAM-1, fibronectin, TNF-alpha and correlated to profile of preeclampsia. The odds ratio (OR) and 95% confidence intervals obtained from unconditional logistic regression were used to set association between TNF-apolymorphism and PE risk. For continuous variables we applied Student's t -test and for categorical variables the Pearson x 2 or Fisher's exact test. The two-way ANOVA with Bonferroni correction was used in multivariate analysis. Results The A allele was more frequent in cases than controls (22.4% vs. 13.2%) what increased disease risk (OR=2.73). Maternal serum levels of TNF-alpha, sVCAM-1 and fibronectin were significantly increased in cases (855.8±385.1pg/mL, 1243±671ng/mL, 0.308±0.231g/L) compared to controls (301.1±156.1pg/mL, 651±250ng/mL, 0.218±0.101g/L, p p p =0.031; respectively) and levels shoved increasing trend with mutant allele in genotype. Moderate and severe proteinuria was higher in rs1800629 allele A subjects compared to G/G carriers (53.8% vs. 14.3%, p p Conclusion This study revealed possible association between clinical and laboratory manifestations of preeclampsia and TNF-alpha gene G308A (rs1800629) polymorphism.

The relationship between TNF alpha gene polymorphisms (− 238/− 308), TNF RII VNTR (p75) and outcomes of hepatitis B virus infection in Tunisian population

The present study was undertaken to investigate the association between Hepatitis B Virus (HBV) infection and polymorphisms of tumour necrosis factor alpha TNF-α −308 G>A, TNF-α −238 G>A and TNF RII VNTR (p75) gene promoter in a Tunisian population. Blood samples were collected from 100 Tunisian patients with HBV infection, 45 with Chronic Hepatitis (CH), 36 with Liver Cirrhosis (LC), 15 with Hepatocellular Carcinoma (HCC) and 200 healthy individuals of similar ethnicity. Genomic DNA was extracted from peripheral blood leukocytes. Genotyping of the analysed polymorphisms was performed using Amplified Refractory Mutation System-Polymerase Chain Reaction (ARMS-PCR), Restriction Fragment Length Polymorphism (RFLP) and Variable Number Tandem Repeat PCR (PCR-VNTR). The variant homozygotes −308 GG were associated with 50% decreased risk of HBV chronic infection (GG vs AA+GA; p=0.010; OR=0.50; 95%CI=0.29–0.85). However, the carriers of minor allele −308 A have higher risk (1.5 times) to develop a chronic infection than other patients (p=0.027; OR=1.46; 95%CI=1.04–2.06). The minor allele of −238 polymorphism was positively associated with virus resistance and the development of chronic infection (p=0.043; OR=1.42; 95%CI =1.01 1.99). The distribution of −308, −238 and TNF RII VNTR (p75) among the three groups differed significantly. For HCC groups, there were statistically significant differences in allele distribution in −308, −238 respectively in which A allele remains a risk factor for HBV evolution to HCC (p=0.008 and p=0.026). Haplotype analysis revealed that TNF-α (−308A; −238A) was significantly associated to HBV chronic infection and moreover to disease aggravation to HCC stage. Our findings imply that variations in the genes governing the levels of constitutive and inducible TNF-α and TNF RII might be an important risk factor, which could explain the variable outcomes of HBV infection.

TNF-alpha gene polymorphisms can help to predict response to etanercept in psoriatic patients.

Genetic factors might have a role for lack of therapeutic response to anti-TNF-alpha agents, as previously suggested in patients with rheumatoid arthritis and inflammatory bowel disease. We evaluated the role of the main TNF-alpha polymorphisms (-238G>A, -308G>A, -857C>T) in predicting the response to etanercept, an anti-TNF-alpha fusion protein. Genomic DNA was extracted from buccal epithelial cells in a series of 97 psoriatic patients who received etanercept for at least 3 months. Patients were classified as responders, if they achieved a PASI improvement ≥ 75% after 12 weeks of etanercept treatment, and non-responders, if PASI improvement was <75%. Single-nucleotide polymorphisms (SNPs) in TNF-alpha gene (-238G>A, -308G>A, -857C>T) were genotyped by PCR restriction fragment length polymorphism (RFLP) assays. We found that patients heterozygous (GA) for the -238G>A polymorphism were more likely not responsive to therapy compared to the GG genotype. In fact, the GA genotype was found in 5/59 (8.5%) responders and in 14/38 (36.8%) non-responders (P = 0.001). A significant relationship with therapy was also observed for the -308G>A polymorphisms. In fact, the GG, GA and AA genotypes were detected in 48 (81.4%), 9 (15.3%) and 2 (3.4%) of the 59 responders and in 22 (57.9%), 11 (28.9%) and 5 (13.2%) of the 38 non-responder patients (P = 0.03). No association with therapy was observed for the -857C>T polymorphisms. Our study supports the role of TNF-alpha polymorphisms in predicting the response to anti-TNF-alpha agents. In particular, we found that the presence of -238G>A and -308G>A polymorphisms is associated with poor response to a 3-month therapy with etanercept. However, our data have yet to be validated in larger cohorts.

Effects of ischemic preconditioning in a pig model of large-for-size liver transplantation

OBJECTIVE:In most cases of pediatric liver transplantation, the clinical scenario of large-for-size transplants can lead to hepatic dysfunction and a decreased blood supply to the liver graft. The objective of the present experimental investigation was to evaluate the effects of ischemic preconditioning on this clinical entity.METHODS:Eighteen pigs were divided into three groups and underwent liver transplantation: a control group, in which the weights of the donors were similar to those of the recipients, a large-for-size group, and a large-for-size + ischemic preconditioning group. Blood samples were collected from the recipients to evaluate the pH and the sodium, potassium, aspartate aminotransferase and alanine aminotransferase levels. In addition, hepatic tissue was sampled from the recipients for histological evaluation, immunohistochemical analyses to detect hepatocyte apoptosis and proliferation and molecular analyses to evaluate the gene expression of Bax (pro-apoptotic), Bcl-XL (anti-apoptotic), c-Fos and c-Jun (immediate-early genes), ischemia-reperfusion-related inflammatory cytokines (IL-1, TNF-alpha and IL-6, which is also a stimulator of hepatocyte regeneration), intracellular adhesion molecule, endothelial nitric oxide synthase (a mediator of the protective effect of ischemic preconditioning) and TGF-beta (a pro-fibrogenic cytokine).RESULTS:All animals developed acidosis. At 1 hour and 3 hours after reperfusion, the animals in the large-for-size and large-for-size + ischemic preconditioning groups had decreased serum levels of Na and increased serum levels of K and aspartate aminotransferase compared with the control group. The molecular analysis revealed higher expression of the Bax, TNF-alpha, I-CAM and TGF-beta genes in the large-for-size group compared with the control and large-for-size + ischemic preconditioning groups. Ischemic preconditioning was responsible for an increase in c-Fos, IL-1, IL-6 and e-NOS gene expression.CONCLUSION:Ischemia-reperfusion injury in this model of large-for-size liver transplantation could be partially attenuated by ischemic preconditioning.

Association of TNF-alpha, IL-6 and IL-1beta gene polymorphisms with polycystic ovary syndrome: a meta-analysis.

BackgroundSeveral studies on the association of TNF-alpha (−308 G/A), IL-6 (−174 G/C) and IL-1beta (−511 C/T) polymorphisms with polycystic ovary syndrome (PCOS) risk have reported conflicting results. The aim of the present study was to assess these associations by meta-analysis.ResultsA total of 14 eligible articles (1665 cases/1687 controls) were included in this meta-analysis. The results suggested that there was no obvious association between the TNF-alpha (−308 G/A) polymorphism and PCOS in the overall population or subgroup analysis by ethnicity, Hardy–Weinberg equilibrium (HWE) in controls, genotyping method, PCOS diagnosis criteria, and study sample size. Also, no obvious association was found between the TNF-alpha (−308 G/A) polymorphism and obesity in patients with PCOS (body mass index [BMI] ≥ 25 kg/m2 vs. BMI < 25 kg/m2). Regarding the IL-6 (−174 G/C) polymorphism, also no association was found in the overall population in heterozygote comparison, dominant model, and recessive model. Even though an allelic model (odds ratio [OR] = 0.63, 95% confidence interval [CI] = 0.41–0.96) and a homozygote comparison (OR = 0.52, 95% CI = 0.30–0.93) showed that the IL-6 (−174 G/C) polymorphism was marginally associated with PCOS. Further subgroup analysis suggested that the effect size was not significant among HWE in controls (sample size ≤ 200) and genotyping method of pyrosequencing under all genetic models. Similarly, there was no association between the IL-1beta (−511 C/T) polymorphism and PCOS in the overall population or subgroup analysis under all genetic models. Furthermore, no significant association was found between the IL-1beta (−511 C/T) polymorphism and several clinical and biochemical parameters in patients with PCOS.ConclusionsThe results of this meta-analysis suggest that the TNF-alpha (−308 G/A), IL-6 (−174 G/C), and IL-1beta (−511 C/T) polymorphisms may not be associated with PCOS risk. However, further case–control studies with larger sample sizes are needed to confirm our results.Electronic supplementary materialThe online version of this article (doi:10.1186/s12863-015-0165-4) contains supplementary material, which is available to authorized users.

Improved inflammatory balance of human skeletal muscle during exercise after supplementations of the ginseng-based steroid Rg1.

The purpose of the study was to determine the effect of ginseng-based steroid Rg1 on TNF-alpha and IL-10 gene expression in human skeletal muscle against exercise challenge, as well as on its ergogenic outcomes. Randomized double-blind placebo-controlled crossover trials were performed, separated by a 4-week washout. Healthy young men were randomized into two groups and received capsule containing either 5 mg of Rg1 or Placebo one night and one hour before exercise. Muscle biopsies were conducted at baseline, immediately and 3 h after a standardized 60-min cycle ergometer exercise. While treatment differences in glycogen depletion rate of biopsied quadriceps muscle during exercise did not reach statistical significance, Rg1 supplementations enhanced post-exercise glycogen replenishment and increased citrate synthase activity in the skeletal muscle 3 h after exercise, concurrent with improved meal tolerance during recovery (P<0.05). Rg1 suppressed the exercise-induced increases in thiobarbituric acids reactive substance (TBARS) and reversed the increased TNF-alpha and decreased IL-10 mRNA of quadriceps muscle against the exercise challenge. PGC-1 alpha and GLUT4 mRNAs of exercised muscle were not affected by Rg1. Maximal aerobic capacity (VO2max) was not changed by Rg1. However, cycling time to exhaustion at 80% VO2max increased significantly by ~20% (P<0.05). Conclusion: Our result suggests that Rg1 is an ergogenic component of ginseng, which can minimize unwanted lipid peroxidation of exercised human skeletal muscle, and attenuate pro-inflammatory shift under exercise challenge.

Investigation of TNF-alpha gene (G308A) and GSTP1 gene (Ilel05Val) polymorphisms in Turkish patients with retinopathy of prematurity.

Retinopathy of prematurity (ROP) is one of the most frequent causes of blindness in newborn babies. Currently, its etiology is not fully understood.-In this study we aimed to investigate the correlation between a patient group with ROP and a control group in terms of the tumor necrosis factor-alpha (TNF-alpha) (G308A) gene and glutathione-S-transferase P1 (GSTP1) (Ilel05Val) gene polymorphism. Sixty-two patients diagnosed with ROP and 58 control subjects were included in this study. For TNF-alpha (G308A) gene and GSTP1 (Ilel05Val) gene polymorphisms, the polymerase chain reaction-restriction fragment length polymorphism method was used. In statistical analysis the significance level was determined as P < 0.05. When the patient and control groups were compared in terms of TNF-alpha (G308A) gene and GSTP1 (Ilel05Val) gene polymorphisms, no statistically significant difference was found (P > 0.05). In our study, no correlation was identified between TNF-alpha (G308A) gene and GSTP1 (Ilel05Val) gene polymorphisms and susceptibility for development of ROP. Further studies are required with more cases of ROP patients and other gene polymorphisms that could be related.

Molecular cloning and expression patterns of two tumor necrosis factor alpha genes in crucian carp (Carassius carassius)

Tumor necrosis factor (TNF) is a potent inflammatory cytokine produced during inflammation. In this study, two Crucian carp TNF-alpha genes, TNFalpha-1 and TNFalpha-2 were cloned and sequenced. The TNFalpha-1 was 720 bp and-consisted of a 699 bp opening reading frame encoding 232 amino acids and TNFalpha-2 was 793 bp and contained an open reading frame of 687 bp which encoded 228 amino acids. The genomic structure of both genes consists of4 exons and 3 introns similar to other known TNF-alpha genes. The amino acid sequence ofcrucian carp TNFalpha-1 and TNFalpha-2 shared the highest identity with common carp, goldfish and the lowest with flounder and human. The phylogenetic analysis grouped crucian carp TNFalpha-1 and TNFalpha-2 with other cypriniformes, which are more closely related to the goldfish and common carp TNFEa-1 and TNFalpha-2. Real time PCR analysis showed a con- stitutive expression of crucian carp TNFalpha-1 and TNFalpha-2 in all seven tissues examined. The TNFalpha-1 mRNA was ex- pressed significantly higher in the liver and kidney than those of TNFalpha-2 while TNFalpha-2 was expressed significantly higher in the muscle than that of TNFalpha-1. Aeromonas hydrophila BSK-10 strain upregulated the expression level of TNFalpha-1 and TNFalpha-2 in all tissues tested. At 6 h, the expression level of TNFalpha-1 increased significantly higher in muscle, skin and liver, while the expression level of TNFalpha-2 increased significantly higher in muscle and gill. TNFalpha-1 expressed much stronger than TNFalpha-2. At 12 h the expression level started declining and was more reduced at 24 h. These results imply that both TNFalpha-1 and TNFalpha-2 mRNA are distributed differently in tissues and are implicated in the immune response to bacterial infection.