Saturated fat ingestion stimulates proatherogenic inflammation in polycystic ovary syndrome

Inflammation and dyslipidemia are often present in Polycystic Ovary Syndrome (PCOS). We determined the effect of saturated fat ingestion on circulating heat shock protein-70 (HSP-70) and mononuclear cell (MNC) toll-like receptor-2 (TLR2) gene expression, activator protein-1 (AP-1) activation and matrix matalloproteinase-2 (MMP-2) protein in women with PCOS. Twenty reproductive-age women with PCOS (10 lean, 10 with obesity) and 20 ovulatory controls (10 lean, 10 with obesity) participated in the study. HSP-70 was measured in serum and TLR2 mRNA and protein, AP-1 activation and MMP-2 protein were quantified in MNC from blood drawn fasting and 2, 3 and 5 hours after saturated fat ingestion. Insulin sensitivity was derived from an oral glucose tolerance test (ISOGTT). Androgen secretion was assessed from blood drawn fasting and 24, 48 and 72 hours after HCG administration. In response to saturated fat ingestion, serum HSP-70, TLR2 gene expression, activated AP-1 and MMP-2 protein were greater in lean women with PCOS compared with lean controls, and in women with PCOS and obesity compared with controls with obesity. Both PCOS groups exhibited lower ISOGTT and greater HCG-stimulated androgen secretion compared with control subjects. Lipid-stimulated proatherogenic inflammation marker responses were negatively correlated with ISOGTT, and positively correlated with abdominal adiposity and HCG-stimulated androgen secretion. In PCOS, saturated fat ingestion stimulates proatherogenic inflammation independent of obesity. This effect is greater when PCOS is combined with obesity compared with obesity alone. Abdominal adiposity and hyperandrogenism may perpetuate proatherogenic inflammation.

Prevalence of Allelic and Genotypic Variants of Il4, Il10, Il12b and Tlr2 Gene Polymorphism in Patients with Chronic Polypoid Rhinosinusitis

Chronic polyposis rhinosinusitis (CPRS) is an important medical and social problem due to its prevalence, combination with other diseases (bronchial asthma, cystic fibrosis), prolonged recurrent course, which leads to a decrease in the quality of life and performance of patients. Bronchopulmonary, rhinogenic orbital and intracranial complications arising against the background of chronic sinusitis not only worsen the quality of life of patients, but also sometimes lead to long-term disability, and sometimes to disability. The results of our study demonstrated that the distribution of the genotypes of polymorphisms of all genes corresponded to the expected, i.e., in this case, the Hardy-Weinberg equilibrium is fulfilled in both groups. Both samples rs2243250 of the IL4 gene were characterized by high frequencies of the unfavorable C allele and the heterozygous T / C genotype, rs3212227 of the IL12B gene of the A allele and the heterozygous A / C genotype, rs1800895 592 C> A in the IL10 gene of the A, C alleles, and heterozygous C rs5743708 in the TLR2 gene of the C allele and the heterozygous C / T genotype, hence the high level of genetic variability of the studied genes.

Prevalence of Allelic and Genotypic Variants of Il4, Il10, Il12b and Tlr2 Gene Polymorphism in Patients with Chronic Polypoid Rhinosinusitis

Chronic polyposis rhinosinusitis (CPRS) is an important medical and social problem due to its prevalence, combination with other diseases (bronchial asthma, cystic fibrosis), prolonged recurrent course, which leads to a decrease in the quality of life and performance of patients. Bronchopulmonary, rhinogenic orbital and intracranial complications arising against the background of chronic sinusitis not only worsen the quality of life of patients, but also sometimes lead to long-term disability, and sometimes to disability. The results of our study demonstrated that the distribution of the genotypes of polymorphisms of all genes corresponded to the expected, i.e., in this case, the Hardy-Weinberg equilibrium is fulfilled in both groups. Both samples rs2243250 of the IL4 gene were characterized by high frequencies of the unfavorable C allele and the heterozygous T / C genotype, rs3212227 of the IL12B gene of the A allele and the heterozygous A / C genotype, rs1800895 592 C> A in the IL10 gene of the A, C alleles, and heterozygous C rs5743708 in the TLR2 gene of the C allele and the heterozygous C / T genotype, hence the high level of genetic variability of the studied genes.

Increased HSP70 and TLR2 Gene Expression and Association of HSP70 rs6457452 Single Nucleotide Polymorphism with the Risk of Chronic Obstructive Pulmonary Disease in the Croatian Population

Heat shock protein 70 (Hsp70) engages Toll-like receptors (TLR) 2 and 4 when found in the extracellular compartment and contributes to inflammation in chronic obstructive pulmonary disease (COPD). Since there is growing evidence for the genetic risk factors for COPD, the gene expression of HSP70, TLR2 and TLR4 was determined, as well as the association between HSP70, TLR2 and TLR4 single nucleotide polymorphisms, (SNPs) and COPD. The gene expression was assessed in peripheral blood cells of 137 COPD patients and 95 controls by a quantitative polymerase chain reaction (qPCR), while a total of nine SNPs were genotyped by TaqMan allelic discrimination real-time PCR. HSP70 and TLR2 gene expression was increased in COPD patients compared to the controls, regardless of the disease severity and smoking status of participants. The rs6457452 SNP of HSP70 was associated with COPD, indicating the protective role of the T allele (OR = 0.46, 95% CI = 0.24–0.89, p = 0.022). Furthermore, COPD C/T heterozygotes showed a decreased HSP70 mRNA level compared to COPD C/C homozygotes. In conclusion, HSP70 and TLR2 may have a role in the pathogenesis of COPD, and the HSP70 rs6457452 variant might influence the genetic susceptibility to COPD in the Croatian population.

The Role of Toll-Like Cell Receptor 2 (Arg753gln) Polymorphism in Allergen Sensitization and Disease Severity in Atopic Dermatitis Patients

Background: Variations in Toll-like receptor 2 (TLR2) encoding gene have been associated with atopic conditions. Objective: The present work aims to analyze single nucleotide polymorphism (SNP) of TLR2 gene Arg753Gln in atopic dermatitis (AD) and its association with allergen sensitization and disease severity. Methodology: 110 AD patients and 75 healthy controls were enrolled and subjected to genotyping of TLR2 gene Arg753Gln by restriction enzyme analysis and allergy investigations. Results: TLR2 Arg753Gln SNP was significantly more frequent in the patients (48%) in comparison to the healthy group (32%) (0R= 1.7). Individuals with the G/A genotype were at a higher risk for AD development by two times. Inhalant allergens specific IgE were distinguished in 80 % of patients with TLR2 gene polymorphism. Conclusion: GA genotype of TLR2 gene is more dominant in severe cases of atopic dermatitis and associated with sensitization to certain inhalant allergens as house dust mites and pollens.

56 Effects of mycotoxins and selected feed additives on viability and tight junction integrity of IPEC-J2 cells

Mycotoxins are deleterious feed contaminants routinely found in cereal gains used in livestock feed. Development of strategies to eliminate mycotoxins and mitigate their harmful effects on the animal is of major importance to the livestock industry. The objective of the current study was to determine the effects of deoxynivalenol (DON), aflatoxin B1 (AFB1) and fumonisin B1(FB1), and selected prebiotic, probiotic, and essential oil blended feed additives, on viability and integrity in porcine intestinal epithelial cell line (IPEC-J2). IPEC-J2 cells were treated with each individual mycotoxin (DON, AFB1 and FB1) with or without additives (Microsecure (MS), Biolex (BL) and Encinnate (EN), Biomatrix Inc., U.S.A). Mycotoxins significantly decreased the trans-epithelial electrical resistance (TEER) of IPEC-J2 cells compared to control with no negative effects on cell viability at 72h. Biolex and EN increased TEER under a DON challenge. The decrease in TEER by AFB1 was minimized in the presence of MS (9.8%), BL (17.1%) and EN (22.8%) relative to AFB1 (P < 0.0001). The protein abundance of claudin 3 was decreased with mycotoxins (P < 0.0001) and this was partially reversed by the additives relative to control cells with additives. In addition, BL and EN significantly increased claudin 4 protein abundance (P = 0.02) when challenged with AFB1. Encinnate significantly increased TLR2 gene expression when challenged with DON (P < 0.05) with MS and BL had a numerical TLR2 gene expression. A numerical increasing of TLR2 gene expression was also observed with feed additives when cells were challenged with AFB1 and FB1. In summary, mycotoxins led to impaired epithelial tight junction integrity and this was in-part rescued by the additives tested through restoration of cellular integrity and TLR-2 dependent immune modulation.

Arg753Gln Polymorphisms in the Toll-Like Receptor 2 Gene are Associated with Cytomegalovirus Infection in Egyptian Bone Marrow Recipients

Background: Previous studies have shown that cytomegalovirus (CMV) induced innate immune response via activation of Toll-like receptor 2 (TLR2). The association between CMV among specific single-nucleotide polymorphisms (SNPs) in the TLR2 gene was also investigated. Objective: This study investigated the relationship between specific SNPs in the TLR2 gene (G>A), TLR2-Arg753Gln (rs5743708), and CMV replication after bone marrow transplantation. Methods: The TLR2-Arg753Gln SNP was genotyped in 181 patients after bone marrow transplantation: 83 and 98 patients with and without CMV infection, respectively. CMV load was determined in serially collected blood samples using real-time PCR. Genotyping was performed using specific sequence primer PCR (SSP-PCR), and the results were confirmed by restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified fragments for GG (wild type), GA and AA identification. Results: Roughly, 85% of the patients screened for the presence of the TLR2-Arg753Gln were GG homozygous, and 15% were GA heterozygous; no patients were homozygous for the mutant allele (A). The GA heterozygous allele was more frequent in the CMV-infected group after bone marrow transplantation. Conclusion: To our knowledge, this is a novel observation that supports the notion that the functional missense mutation (TLR2-Arg753Gln polymorphism) is possibly associated with CMV replication after bone marrow transplantation. This suggests a role for TLR2 in the innate immune response of human CMV infection in Egyptian bone marrow recipients..