RESUMEN

Bovine mastitis is a complex infectious disease that develops in the mammary gland, predominantly caused by a bacterial infection of mammary tissue. Genetic variability of mastitis is well established and depends upon different quantitative trait loci (QTL) related to mastitis resistance or susceptibility. The susceptibility is often attributed to single-nucleotide polymorphisms (SNPs) in the variable cow breed genomes. Several global investigative attempts have resulted in studies mapping mastitis to the variations in the relevant genes. Reports have been attributed to dramatic genetic expression changes in Toll-Like Receptor 4 (TLR4) genes in mastitis-positive cows. However, the mechanism behind this variable genetic expression of TLR4 genes has been studied poorly. The present study aims to investigate SCM through various screening tests like somatic cell count (SCC), electric conductivity (EC), pH, and California mastitis test (CMT) in milk samples. This study also aims to investigate possible mechanisms behind this variable expression of TLR4 by comparative SNP evaluation and transcriptional factor profile mining. So that the important genetic mutations and effects thereof can be exploited in selecting specific breeds with higher mastitis resistance and milk yield. Seventy Holstein Frisian (HF) crossbred dairy cows were selected in the present study. The animals were screened based on various diagnostic tests (SCC, pH, EC, and CMT). Blood samples (5 mL) were collected for extraction of DNA followed by amplification of PPR1 and PPR2 of the promoter region and 5'UTR of the bovine TLR4 gene using specific primers. Sanger's enzymatic DNA sequencing technique sequenced the amplified PCR products. Further, the identification of SNPs was done through various bioinformatic tools used in this study. The findings of the present study revealed that CMT, EC, pH, and SCC could be used for the early detection of subclinical mastitis. In the present study, a significant increase in the EC, pH, and SCC in milk samples of animals affected with SCM was found in comparison to the healthy animals. The present study also revealed 16 SNPs falling in TLR4 promoter and 5' untranslated region (5'UTR) sequences in mastitis-positive genotypes compared to reference genomes. The study also investigates the potential transcriptional factor program deployed in response to variable mastitis development resistance. In the present study, the allelic and genotype frequencies of all SNP variants in the three regions viz., PPR1, PPR2, and 5'UTR, were the same indicating the absence of heterozygous condition at the respective loci. The present study has wide applicability for researchers developing mastitis-resistant breeding programs and the data generated may aid in the selection of better genetic breeds. The transcription factor binding profiles can serve as concrete leads about the studies on bovine mastitis at the molecular level and may also aid global research groups working on transcription factor (TF)-based molecular pathology of mastitis.

RESUMEN

Single-nucleotide polymorphisms (SNPs) in the Toll-like receptor 4 (TLR4) gene have been documented in type 2 diabetes mellitus (T2DM) and other diseases in the Saudi population. We investigated the relationship between rs11536889, rs4986790, and rs4986791 SNPs in the TLR4 gene and T2DM in the Saudi population; 105 patients with T2DM and 105 healthy controls were analyzed. The TLR4 gene was amplified through PCR, followed by restriction fragment length polymorphism analysis for rs4986791 and Sanger sequencing for rs11536889 and rs4986790 SNPs. The clinical and biochemical characteristics were associated with T2DM (p < 0.05). The rs11536889, rs4986790, and rs4986791 SNPs in control subjects followed the Hardy-Weinberg equilibrium (p > 0.05). Alleles were associated with rs11536889, rs4986791, heterozygous codominant, and dominant models (p < 0.05). However, the rs4986790 SNP was not associated with T2DM (p > 0.05). Logistic regression analysis showed that high-density lipoprotein cholesterol (HDLc) levels were associated with T2DM (p < 0.001). Analysis of variance showed that waist (p = 0.0005) and hip circumferences (p = 0.002) in rs4986790 and rs4986791 SNPs, in SBP (p = 0.001), DBP (p = 0.002), and HDLc levels (p = 0.003), were associated with T2DM subjects. T2DM was also associated with the haplotype (p < 0.001) but not with linkage disequilibrium. The gene-gene interaction was associated with the three SNPs studied in patients with T2DM according to the generalized multifactor dimensionality reduction model (p < 0.0001). Dendrogram and graphical depletion analysis revealed a moderate association in patients with T2DM. The results suggest that rs11536889 and rs4986790 SNPs are genotypically and allelically associated with T2DM in Saudi patients. Future functional studies are recommended to validate the genetic roles of these SNPs in the pathogenesis and progression of diseases.

Asunto(s)

Diabetes Mellitus Tipo 2 , Receptor Toll-Like 4 , Humanos , Receptor Toll-Like 4/genética , Predisposición Genética a la Enfermedad , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/complicaciones , Arabia Saudita , Alelos

RESUMEN

Introduction: The aim of the present study was to investigate the association between the single nucleotide polymorphism (SNP) rs1927914 A/G in TLR4 gene and the immunological profile of household contacts (HHC) of leprosy patients. Leprosy classification is usually complex and requires the assessment of several clinical and laboratorial features. Methods: Herein, we have applied distinct models of descriptive analysis to explore qualitative/quantitative changes in chemokine and cytokine production in HHC further categorized according to operational classification [HHC(PB) and HHC(MB)] and according to TLR4SNP. Results and discussion: Our results showed that M. leprae stimuli induced an outstanding production of chemokines (CXCL8;CCL2; CXCL9; CXCL10) by HHC(PB), while increase levels of pro-inflammatory cytokines (IL-6; TNF; IFN-γ; IL-17) were observed for HHC(MB). Moreover, the analysis of chemokine and cytokine signatures demonstrated that A allele was associated with a prominent soluble mediator secretion (CXCL8; CXCL9; IL-6; TNF; IFN-γ). Data analysis according to TLR4 SNP genotypes further demonstrated that AA and AG were associated with a more prominent secretion of soluble mediators as compared to GG, supporting the clustering of AA and AG genotypes into dominant genetic model. CXCL8, IL-6, TNF and IL-17 displayed distinct profiles in HHC(PB) vs HHC(MB) or AA+AG vs GG genotype. In general, chemokine/cytokine networks analysis showed an overall profile of AA+GA-selective (CXCL9-CXCL10) and GG-selective (CXCL10-IL-6) axis regardless of the operational classification. However, mirrored inverted CCL2-IL-10 axis and a (IFN-γ-IL-2)-selective axis were identified in HHC(MB). CXCL8 presented outstanding performance to classify AA+AG from GG genotypes and HHC(PB) from HHC(MB). TNF and IL-17 presented elevated accuracy to classify AA+AG from GG genotypes and HHC(PB) (low levels) from HHC(MB) (high levels), respectively. Our results highlighted that both factors: i) differential exposure to M. leprae and ii) TLR4 rs1927914 genetic background impact the immune response of HHC. Our main results reinforce the relevance of integrated studies of immunological and genetic biomarkers that may have implications to improve the classification and monitoring of HHC in future studies.

Asunto(s)

Lepra , Mycobacterium leprae , Humanos , Interleucina-17 , Receptor Toll-Like 4/genética , Interleucina-6 , Citocinas , Lepra/genética , Inmunidad , Quimiocinas

RESUMEN

BACKGROUND: The single nucleotide polymorphisms in the TLR4 gene can decrease or increase the response to lipopolysaccharide, increasing the susceptibility to inflammatory diseases, affecting the expression or receptor function by inducing a low-grade chronic inflammatory response. PURPOSE: The objective of this study was to evaluate the association of SNPs - 2570 A > G (rs2737190), - 2081 G > A (rs10983755), 896 A > G (rs 4986790), and 1196 C > T (rs4986791) of the TLR4 gene with obesity and metabolic alterations in the young population. RESULTS: In this study, it was found that the carriers of the heterozygous genotype of the SNPs - 2081 G > A, 896 A > G, and 1196 C > T confer a higher risk of developing obesity (OR = 3.73, p = 0.018; OR = 5.66, p = 0.014, and OR = 8.95, p = 0.014, respectively). Also, with the lipid profile, the SNP - 2081 G > A was associated with total cholesterol (TC) ≥ 200 mg/dL (OR = 3.91, p = 0.020) and Kannel index > 3% (OR = 4.00, p = 0.008). The SNP 896 A > G was associated with LDL-c ≥ 100 mg/dL (OR = 3.64, p = 0.040) and Kannel index > 3% (OR = 4.33, p = 0.016), and the SNP 1196 C > T was associated with TC ≥ 200 mg/dL (OR = 4.37, p = 0.048), Castelli index > 4.5/> 5% (OR = 5.33, p = 0.016), and Kannel index > 3% (OR = 16.00, p = 0.001). Finally, the AGGT haplotype was associated with Castelli index > 4.5/> 5% (OR = 5.40, p = 0.015) and Kannel index > 3% (OR = 10.46, p < 0.001), and the AAAC haplotype was associated with obesity (OR = 3.56, p = 0.020), TC ≥ 200 mg/dL (OR = 4.04, p = 0.007), LDL-c ≥ 100 mg/dL (OR = 2.98, p = 0.030) and Kannel index > 3% (OR = 4.20, p = 0.002). CONCLUSION: The heterozygous genotype of the SNPs - 2081 G > A, 896 A > G and 1196 C > T of the TLR4 gene was associated with altered lipid profile and development of obesity in young university students of Guerrero State, Mexico.

Asunto(s)

Obesidad , Receptor Toll-Like 4 , Humanos , Haplotipos , Receptor Toll-Like 4/genética , Proyectos Piloto , LDL-Colesterol , Genotipo , Obesidad/epidemiología , Obesidad/genética , Polimorfismo de Nucleótido Simple , Predisposición Genética a la Enfermedad

RESUMEN

Objective:To explore the effect of triglyceride glucose(TyG) index, single nucleotide polymorphism of Toll-like receptor 4(TLR4) and NOD-like receptor thermal protein domain associated protein 3(NLRP3) genes, and its interaction on the risk of gout.Methods:A total of 315 male patients with gout and 499 men for health checkup at the same period were selected. General data were collected through questionnaires, and peripheral venous blood was collected for biochemical test. Three single nucleotide polymorphisms(SNPs) of NLRP3 and TLR4 were detected with multiplex ligase assay reaction, and logistic regression analysis was applied to compare the correlation between NLRP3 and TLR4 alleles and gout risk. The interaction of SNP and TyG index with gout was analyzed by generalized multi-factor dimensionality reduction(GMDR) model and logistic regression.Results:After adjusting for smoking, drinking, and other factors, the risk of gout increased by 61.1% for each standard deviation increase in TyG index. CC genotypes of rs10754558, rs10759932, and rs7525979 were high risk genotypes of gout in Han ethnicity. GMDR results showed significant differences in the interaction models of rs10754558-TyG index, rs7525979-TyG index, and rs10759932-TyG index between control group and gout group( P<0.05), suggesting an interaction between the three genotypes of SNPs selected and TyG index. Stratified analysis of the three selected SNPs and TyG index showed that after adjusting for age, smoking, and other factors, the high TyG index patients carrying C/C or C/G genotype at rs10754558 displayed an increased risk of gout compared with those carrying GG genotype and low TyG index( OR=2.127, P<0.05). Conclusion:The CC genotypes of rs10754558, rs10759932, and rs7525979 are high risk genotypes for gout in Han ethnicity. The interaction between rs10754558 and TyG index may increase the risk of gout development.

RESUMEN

Coccidiosis caused by Eimeria spp. is a protozoan disease prevalent in farm animals, and it is responsible for serious economic losses especially in young animals. It has been popular to breed disease-resistant animals due to the concern about food safety, animal welfare, and public health. Toll-like receptor (TLR) gene family plays a key role in the innate immune system participating in host-antigen interaction, therefore, they are candidate genes for breeding disease-resistant animals. In the present study, possible genetic associations between TLR4 gene coding variants and the presence of Eimeria spp. in adult Turkish sheep were investigated. For this purpose, the presence of Eimeria spp. in fecal samples from six native Turkish sheep were determined, and approximately 1450 bp region in the 3rd exon of the ovine TLR4 gene was sequenced. Ten nonsynonymous and four synonymous single nucleotide polymorphisms (SNPs) were detected in the targeted region. Statistical analyses revealed that the SNP at the codon at 356th position encoding Leucine instead of Phenylalanine (F356L) was significantly associated with the presence of Eimeria spp. It was found that the individuals carrying at least one Leucine amino acid sequence at this position have 2.3-fold more risk for the presence of Eimeria spp.

Asunto(s)

Coccidiosis/veterinaria , Eimeria , Enfermedades de las Ovejas/parasitología , Receptor Toll-Like 4/metabolismo , Animales , Coccidiosis/epidemiología , Coccidiosis/parasitología , Heces/parasitología , Predisposición Genética a la Enfermedad , Variación Genética , Ovinos , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/genética , Receptor Toll-Like 4/genética , Turquía/epidemiología

RESUMEN

In this study, the yak's TLR4 gene alternative spliceosomes were investigated using PCR amplification and cloning to improve disease-resistance in yak and promote efficient utilization of yak's resources. qRT-PCR was used to determine the expression levels of two alternatively spliced transcripts of the TLR4 gene in seven distinct tissues. To predict the function of proteins expressed by each TLR4 spliceosome, bioinformatic analysis of yak's TLR4 protein structure and function was performed, which led to the identification of two alternative spliceosomes of yak's TLR4 gene. The TLR4-X1 sequence length was 2526 bp, and it encoded full-length TLR4 protein (841 amino acids). The sequence length of the exon-2 deleted TLR4-X2 sequence was 1926 bp, and it encoded truncated TLR4 protein (641 amino acids). TLR4-X2 sequence was consistent with the predicted sequence of the TLR4 gene in GenBank. Each tissue showed significantly different expression levels of these two alternative spliceosomes. As per the bioinformatic analysis of the structure and function of TLR4 protein, deletion of exon-2 in the TLR4 gene resulted in frameshift mutations of the reading frame in the corresponding protein, which altered its ligand-binding and active sites. Besides, biological property such as substrate specificity of truncated TLR4 protein was also altered, leading to altered protein function. This study has laid a theoretical foundation for exploring the role of two variants of the TLR4 gene in yak's disease resistance. Besides, this study's data could be analyzed further to explore the molecular mechanism associated with disease-resistance in the yak.

RESUMEN

BACKGROUND: TLR4 is involved in H. pylori lipopolysaccharide recognition and its SNPs might be related to increased risk of developing premalignant conditions and gastric cancer. The objectives of the study were to evaluate the associations between both TLR4 rs4986790 and rs4986791 gene polymorphisms and H. pylori infection in children with gastritis. METHODS: We performed a cross-sectional study on 150 children admitted in a Tertiary Centre from Romania, between March 2016 and July 2018 in order to evaluate them regarding demographic, endoscopic, histopathological and TLR4 gene polymorphisms. RESULTS: Our final sample consisted of 50 children with H.pylori associated gastritis (group 1-Ghp group) and 97 children with gastritis without H.pylori infection (group 2). Poor socioeconomic status was a significant risk factor for H.pylori infection. We found no significant differences regarding the clinical symptoms and laboratory parameters between the two groups. Concordant results were found between the histopathological exam and rapid urease test. Variant genotypes of TLR4rs4986790 and TLR4rs4986791 gene polymorphisms acted as protective factors against H. pylori infection, without statistical significance. CONCLUSIONS: The variant genotype of the TLR4 gene polymorphisms might be protective factors for H.pylori infection, while socioeconomic status is an risk factor for H. pylori infection. Urease test is a usefull diagnostic tool for H. pylori infection.

Asunto(s)

Gastritis/genética , Infecciones por Helicobacter/genética , Helicobacter pylori/patogenicidad , Polimorfismo de Nucleótido Simple , Receptor Toll-Like 4/genética , Adolescente , Edad de Inicio , Niño , Preescolar , Estudios Transversales , Femenino , Gastritis/epidemiología , Gastritis/inmunología , Gastritis/microbiología , Predisposición Genética a la Enfermedad , Infecciones por Helicobacter/epidemiología , Infecciones por Helicobacter/inmunología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/inmunología , Interacciones Huésped-Patógeno , Humanos , Lactante , Masculino , Estudios Prospectivos , Factores Protectores , Medición de Riesgo , Factores de Riesgo , Rumanía/epidemiología , Determinantes Sociales de la Salud , Factores Socioeconómicos , Receptor Toll-Like 4/inmunología

RESUMEN

OBJECTIVE: The aim of the research is to study the prevalence and to determine the prognostic significance of polymorphism ARG753GLN of the TLR-2 gene, Leu412Phe of TLR-3, Asp299Gly of TLR-4 in influenza. PATIENTS AND METHODS: Materials and methods: 112 patients with influenza were examined (63 patients with uncomplicated course and 49 with influenza-associated pneumonia). The genotyping of the polymorphic site of ARG753GLN of the TLR-2 gene, Asp299Gly of the TLR-4 gene, and Leu412Phe of the TLR-3 gene was carried out by polymerase chain reaction using oligonucleotide primers. RESULTS: Results: It has found that the prevalence of the mutant allele 299Gly of TLR-4 in patients with uncomplicated influenza is 6.4 %, with influenza- associated pneumonia - 7.1 %, which exceeds the population control indicators by 3.8-4.3 times (1.7 %, p<0.05). Mutant allele 412Phe of TLR-3 is significantly more common in patients with influenzaassociated pneumonia (42.9%), as compared with uncomplicated influenza (24.6%, p<0.01) and healthy people (30.0%, p<0.05). The increased risk of influenza development is associated with the Asp/Gly genotype of TLR-4 (OR=4.22) and combination of mutant genotypes Leu/Phe and Phe/Phe of TLR-3 with Asp/Gly of TLR-4 and Arg/Gln of TLR-2 (OR=15.0); influenza-associated pneumonia - with genotype Phe/Phe of TLR-3 (OR=4.5). CONCLUSION: Conclusions: It has been found out that among patients with influenza and influenza-associated pneumonia, the mutant allele 299Gly of TLR-4 and combinations of polymorphisms Arg753Gln of TLR-2, Leu412Phe of TLR-3, Asp299Gly of TLR-4 are detected reliably more often. The frequency of the mutant allele 412Phe of TLR-3 is higher among patients with influenza-associated pneumonia. Markers of increased risk of influenza are 299Gly allele and genotype Asp/Gly of TLR-4 and the combination of mutant genotypes Leu/Phe and Phe/Phe of TLR-3 with Asp/Gly of TLR-4 and Arg/Gln of TLR-2; for influenza-associated pneumonia - allele 412Phe and genotype Phe/Phe of TLR-3.

Asunto(s)

Gripe Humana , Neumonía , Polimorfismo Genético , Receptor Toll-Like 2/genética , Receptor Toll-Like 3/genética , Receptor Toll-Like 4/genética , Estudios de Casos y Controles , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Gripe Humana/genética , Neumonía/genética

RESUMEN

OBJECTIVE: Introduction: HIV-infection and chronic hepatitis C is of great concern of current infectology. The paper is aimed at the study of the prevalence of the of TLR4 gene Gly and TLR7 gene Leu polymorphic alleles among patients with HIV/HCV-coinfection in general and with regard to gender, as well as to determine their role in the development of the infection. PATIENTS AND METHODS: Materials and methods: To achieve the objective of the research a cohort and case-control study has been carried out. The total of 535 people has been examined, including: HIV/HCV-coinfected - 104, HIV-monoinfected - 90, patients with chronic hepatitis C - 166 and almost healthy people (population control group) - 175 subjects. RESULTS: Results and conclusion:The study found thatthe prevalence of the TLR4 gene Gly polymorphic allele among patients with HIV/HCV-coinfection, HIV-monoinfection and chronic hepatitis C accounted for 23.1 %, 14.4 % and 14.5 %, respectively, which is significantly higher than the similar index in controls - 3.3 %. The presence of the TLR4 gene Gly polymorphic allele in the genome increases the risk of HIV/HCV-coinfection development, in case of infection, by 9 (OR=8.70, р=0.000), HIV-monoinfection and chronic hepatitis C by 5 times (OR=4.89, р=0,016 and OR=4.9, р=0.011, respectively). TLR7 gene Leu polymorphic allele is recorded with the frequency of 19.9-26.0 % in patients with HIV/HCV-coinfection, HIV-monoinfection and chronic hepatitis C as compared to controls (25.9 %). In female patients with HIV/HCV-coinfection, HIV-monoinfection, chronic hepatitis C and healthy individuals the TLR7 gene Leu polymorphic allele is recorded by 2.1-3.6 times more frequently than in male patients.

Asunto(s)

Coinfección , Infecciones por VIH/genética , Hepatitis C Crónica/genética , Receptor Toll-Like 4/genética , Receptor Toll-Like 7/genética , Estudios de Casos y Controles , Coinfección/genética , Coinfección/virología , Femenino , Humanos , Masculino , Polimorfismo Genético , Prevalencia

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Abstract This study investigated the relationship of polymorphismsin genes encoding CD14, IL-6 and TLR4 withmetabolic, inflammatory and endothelial markers in youngadults with acute myocardial infarction (AMI). Glucose,lipids, nitrate and inflammatory markers, flow mediatedvasodilatation (FMV) and flow mediated by nitrate (FMN)were evaluated in 102 AMI and 108 non-AMI (controlgroup) young individuals (% years). CD14 -260C[T(rs2569190), IL6 -174G[C (rs1800795) and TLR4c.896A[G (rs4986790) and TLR4 c.1196C[T (rs4986791)polymorphisms were analyzed by PCR–RFLP. Minor allelefrequencies of CD14, IL6 and TLR4 polymorphisms weresimilar between AMI and control groups (p[0.05). In AMIgroup, individuals carrying IL6 -174CC genotype hadhigher serum triglycerides, VLDL cholesterol and glucosecompared to the IL6 -174GG/GC genotype carriers(p84 2013-11-12

Asunto(s)

Infarto del Miocardio , Polimorfismo Genético , Receptores de Lipopolisacáridos

RESUMEN

BACKGROUND: To analyze the association between TLR-4 Asp299Gly and Thr399Ile gene polymorphisms and chronic periodontitis in a sample of south Indian population. MATERIALS AND METHODS: Genomic DNA was obtained from peripheral blood of 60 patients with chronic periodontitis and 60 periodontally healthy subjects. TLR-4 Asp299Gly and Thr399Ile gene polymorphisms were genotyped by a polymerase chain reaction-restriction fragment length polymorphism method. The data were analyzed by a χ(2)-test and by relative risk estimation. RESULTS: Thr399Ile alleles were found in 4% of chronic periodontitis patients and in 1% of periodontally healthy subjects. The prevalence of a Thr399Ile heterozygote was found to be 5% in the chronic periodontitis group and 1.67% in the periodontally healthy group, respectively. Homozygosity for TLR-4 Thr399Ile was seen in chronic periodontitis patients only, which was 1.67%. The TLR-4 Asp299Gly gene polymorphism was not detected in either chronic periodontitis or periodontally healthy groups. CONCLUSION: There is no significant association between TLR-4 Thr399Ile polymorphism and chronic periodontitis in a sample of south Indian population.