TLR4 Gene Research Articles

THE ROLE OF ANTIBACTERIAL THERAPY IN THE TREATMENT OF ATOPIC DERMATITIS IN CHILDREN

Background. Atopic dermatitis is one of the most common chronic inflammatory skin diseases. In atopic dermatitis, the diversity of the normal microflora decreases, so that the number of S. aureus prevails and the number of bacteria with antistaphylococcal activity decreases. The aim of this study was to evaluate the effectiveness of mupirocin 2% cream in the treatment of atopic dermatitis in children as an additional therapy, and the effect of the rs4696480 polymorphism in the TLR2 gene on the effectiveness of the treatment.Materials and methods. The study included patients with atopic dermatitis (n = 37), aged 1-18 years (7.9±4.9) from the Department of Allergy at Kyiv City Children Clinical Hospital №2. All children included in the study with atopic dermatitis had a positive S. aureus culture. Patients were randomized into two groups: a group receiving mupirocin 2% cream on the affected skin areas 2 times a day 10 days (group A) and a control group (group B). Children of the control group received only symptomatic therapy. The SCORAD score and CDLQI (children's dermatology life quality index) questionnaire score was recorded before and after the treatment, side effects were recorded during the study. Skin swab cultures were taken before and after treatment. Genotyping for TLR2 rs4696480 was performed by using Real-time PCR.This study was approved by the ethical committee of the O.Bogomolets National Medical University; all patients/parents of the children gave an informed consent to participate. Statistical processing of the obtained data was carried out using the statistical package IBM SPSS Statistics Base (version 22) and the software EZR version 1.32 (graphical interface of the R environment (version 2.13.0). The difference in the effectiveness of therapy between the two subgroups and the influence of genotype on the effectiveness of therapy was determined using the Student criterion for parametric data (T) and Wilcoxon W-test for nonparametric data (W). The dynamics of indicators in each group before and after treatment were evaluated using the Wilcoxon T-test (T-W). Eeffectiveness of S. aureus eradication was estimated using the odds ratio (OR) with a 95% confidence interval (CI). Results were considered statistically significant at the level of p < 0.05.Results. In both groups of children, improvement was observed in 10 days after the treatment. In group A, the improvement in ΔSCORAD was found to be 13.8 (T-W=190.0, p<0.001). In group B, there was also an improvement: ΔSCORAD 8.5 (T-W=153.0, p<0.001). The difference in ΔSCORAD scores between groups A and B was statistically significant (T=2.70, p=0.011). The decrease in CDLQI score after treatment was 31.3% in group A (T-W=190.0, p<0.001), and 18.3% in group B (T-W=171.0, p<0.001). The difference between these two groups was not statistically significant (W=334.0, p=0.409).After the treatment, the skin culture showed that in group A, 57.9% of patients were S. aureus-negative, in group B, only 22.2% of children had a negative culture (OR= 5.50, CI 1.32-22.86).We compared ΔSCORAD depending on the genotype of the rs4696480 polymorphism in TLR2 gene and found no difference in the two subgroups: In the subgroup with the AA genotype, ΔSCORAD was found to be 12.6±3.7; in the subgroup with the TT genotype ‒ 14.4±5.1 (T=0.84, p=0.413).Conclusions. Our results demonstrate the effectiveness of the use of topical mupirocin in the treatment of atopic dermatitis as an additional therapy.

Paeoniflorin alleviates inflammation in bovine mammary epithelial cells induced by Staphylococcus haemolyticus through TLR2/NF-κB signaling pathways

Staphylococcus haemolyticus (S. haemolyticus) is one of the most common coagulase-negative staphylococci (CoNS) isolates from bovine mastitis. Paeoniflorin (PF) shows anti-inflammatory effects on different inflammatory diseases in vitro studies and in vivo animal experiments. In this study, the viability of bovine mammary epithelial cells (bMECs) was detected by the cell counting kit-8 experiment. Subsequently, bMECs were induced with S. haemolyticus, and the induction dosage was determined. The expression of pro-inflammatory cytokines and toll-like receptor (TLR2) and nuclear factor kappa-B (NF-κB) signaling pathway-related genes were investigated by quantitative real-time PCR. The critical pathway proteins were detected by western blot. The results showed that the multiplicity of infection (MOI; the ratio of bacteria to bMECs) 5:1 of S. haemolyticus for 12 h could cause cellular inflammation, which was selected to establish the inflammatory model. Incubation with 50 μg/ml PF for 12 h was the best intervention condition for cells stimulated by S. hemolyticus. Quantitative real-time PCR and western blot analysis showed that PF inhibited the activation of TLR2 and NF-κB pathway-related genes and the expression of related proteins. Western blot results showed that PF suppressed the expression of NF-κB unit p65, NF-κB unit p50, and MyD88 in bMECs stimulated by S. haemolyticus. The inflammatory response pathway and molecular mechanism caused by S. haemolyticus on bMECs are related to TLR2-mediated NF-κB signaling pathways. The anti-inflammatory mechanism of PF may also be through this pathway. Therefore, PF is expected to develop potential drugs against CoNS-induced bovine mastitis.

Molecular characterization of a teleost-specific toll-like receptor 22 (tlr22) gene from yellow catfish (Pelteobagrus fulvidraco) and its transcriptional change in response to poly I:C and Aeromonas hydrophila stimuli

Toll-like receptors (TLRs) are a class of pattern recognition receptors (PRRs) that can recognize pathogen-associated molecular patterns (PMPs) and play important roles in the innate immune system in vertebrates. In this study, we identified a teleost-specific tlr22 gene from yellow catfish (Pelteobagrus fulvidraco) and its immune roles in response to different pathogens were also determined. The open reading frame (ORF) of the tlr22 was 2892 bp in length, encoding a protein of 963 amino acids. Multiple protein sequences alignment, secondary and three-dimensional structure analyses revealed that TLR22 is highly conserved among different fish species. Phylogenetic analysis showed that the phylogenetic topology was divided into six families of TLR1, TLR3, TLR4, TLR5, TLR7 and TLR11, and TLR22 subfamily was clustered into TLR11 family. Meanwhile, synteny and gene structure comparisons revealed functional and evolutionary conservation of the tlr22 gene in teleosts. Furthermore, tlr22 gene was shown to be widely expressed in detected tissues except barbel and eye, with highest expression level in liver. The transcription of tlr22 was significantly increased in spleen, kidney, liver and gill tissues at different timepoints after Poly I:C infection, suggesting TLR22 plays critical roles in defensing virus invasion. Similarly, the transcription of tlr22 was also dramatically up-regulated in spleen, kidney and gill tissues with different patterns after Aeromonas hydrophila infection, indicating that TLR22 is also involved in resisting bacteria invasion. Our findings will provide a solid basis for the investigation the immune functions of tlr22 gene in teleosts, as well as provide useful information for disease control and treatment for yellow catfish.

Myrrh Essential Oil Mitigates Renal Ischemia/Reperfusion-Induced Injury.

Ischemia/reperfusion (I/R)-induced renal injury is a common reason for kidney injury in clinical settings; therefore, continuous investigation of novel nephroprotective agents is crucial. Myrrh, the oleoresin exudates generated by the genus Commiphora, display numerous pharmacological actions. This study tried to assess the preventive effects of myrrh essential oil against I/R-induced renal damage. Rats were randomized into five groups. In the sham group, the animals were subjected to bilateral renal artery separation with no occlusion. In the sham + myrrh group; the rats were administered myrrh essential oil and then treated similarly to the sham group. Renal I/R group: the animals were challenged with renal I/R. In the myrrh + renal I/R groups, rats were administered 50 or 100 mg/kg of myrrh essential oil orally for three weeks before being confronted with I/R. Serum levels of renal function tests and renal injury biomarkers, including NGAL, KIM-1, and CysC, were amplified in the renal I/R group. Animals that experienced renal I/R exhibited elevated lipid peroxidation (MDA); declined SOD, CAT, and GPx activity; declined GSH content; augmented TLR4/NFκB gene expression; and subsequent enhancement of inflammatory mediators (TNF-α, IFN-γ, IL-1β, and IL-6). Myrrh reduced renal function tests and injury biomarkers and amended renal histological alterations. Pretreatment with myrrh reduced MDA, elevated the antioxidant enzymes' activities and GSH content, and reduced the TLR4 and NFκB gene expression, leading to subsequent inflammation and apoptosis alleviation. The outcomes of the present investigation established the protective effect of myrrh essential oil against renal I/R via pointing out the antioxidant, anti-inflammatory, and anti-apoptotic effects of myrrh.

Association of TLR4 and TLR9 Gene Polymorphisms with the Risk and Progression of Cervical Lesions in HPV-Infected Women

Aim: Toll-like receptors involved in tumor-associated inflammatory response, this study aimed to investigate the role of TLR4 and TLR9 gene polymorphisms in the risk and progression of HPV-related cervical lesions. Materials & methods: A total of 220 cervical lesion patients and 227 healthy controls were enrolled. Single-nucleotide polymorphisms were genotyped using PCR-restriction fragment length polymorphism. Results: A significantly decreased risk of cervical lesions was observed to be associated with the TLR4 rs10116253 (C), rs1927911 (T) and rs10759931 (G) mutant alleles. rs187084-rs1927911-HPV-16/18 was the best interaction model to affect cervical lesion risk. Conclusion: TLR4 rs10116253, rs1927911 and rs10759931 were potential biomarkers for cervical lesion susceptibility.

AdipoRon Effect on Expression of Lipid Metabolism Genes in Cultured Human Primary Macrophages

Atherosclerosis is characterized by excessive uptake of cholesterol-rich low-density lipoprotein (LDL) by vascular wall macrophages. The macrophages are transformed into foam cells, lipids accumulate in the intima of arteries, atherosclerotic plaques arise, and cardiovascular diseases develop. Adiponectin is an adipose tissue adipokine and possess anti-atherogenic and anti-inflammatory activities, which are mediated by adiponectin binding to its receptors AdipoR1 and AdipoR2. To exert its anti-atherogenic effect, adiponectin may regulate the reverse cholesterol transport and prevent foam cells formation. The small-molecule adiponectin receptor agonist AdipoRon was assumed to modulate expression of reverse cholesterol transport and inflammation genes in human macrophages. Several AdipoRon concentrations (0, 5, 10, and 20 μM) were tested for effect on expression of the lipid metabolism genes ABCA1, ABCG1, APOA1, NR1H3 (LXRα), NR1H2 (LXRβ), PPARG, and ACAT1 and the inflammation genes IL6, TNFA, and TLR4 in cultured human primary macrophages and the THP-1 macrophage cell line. Cell viability was measured using the MTS assay. ABCA1, ABCG1, APOA1, NR1H3, NR1H2, PPARG, ACAT1, IL6, TNFA, and TLR4 mRNA levels in human primary macrophages were assessed by real-time PCR. The PPARG and ABCA1 relative mRNA levels were found to increase in human primary macrophages treated with 5 or 10 μM AdipoRon for 24 h. A higher AdipoRon concentration (20 μM) was cytotoxic to macrophages, especially THP-1 cells. The effect of AdipoRon on human macrophages and potential adiponectin receptor agonists are of interest to study in view of the need to develop new approaches to atherosclerosis prevention and treatment.

Curcumin interferes with sepsis-induced cardiomyocyte apoptosis via TLR1 inhibition

ObjectiveSepsis-induced cardiomyopathy is the leading cause of death in sepsis and is characterized by reversible myocardial depression. However, the specific mechanisms responsible for myocardial injury in sepsis are not known. The present study used bioinformatic analysis to explore the possible mechanisms of sepsis-induced myocardial injury and the therapeutic potential of curcumin. MethodsThe GSE125042 microarray gene expression matrix was obtained from the Gene Expression Omnibus database, which includes 10 septic cardiomyocyte samples from cecum ligation perforation constructs and 10 sham-operated groups cardiomyocyte samples. Background correction and matrix data normalization were performed using the robust multiarray average algorithm. Differentially expressed genes (DEGs) screening was performed using the Limma R package expression matrix, and whole gene analysis was performed using the weighted gene co-expression network analysis R package to construct gene networks and identify modules. Enrichment analysis and gene set enrichment analysis was performed on the genes to be selected. Construct cellular and animal models of myocardial injury in sepsis were assessed and the effects of curcumin on a rat or cardiac myocytes were observed. ResultsA total of 2876 DEGs were screened based on the GSE125042 chip, of which 1424 genes were upregulated and 1452 genes were down regulated. WGCNA analysis of the whole genes was also performed and a total of 20 gene modules were generated. Among them, the selected TLR1 gene was present in the most strongly correlated Brown module. Enrichment analysis of the upregulated DEGs with the Brown module showed that they were significantly enriched in biological processes related to ribosomal protein complex generation, cellular components related to phagocytic vesicles and molecular functions related to Toll-like receptor binding, affecting cardiomyocyte survival as a target for molecular intervention in septic cardiomyopathy. Animal experiments showed that curcumin reduced inflammation levels, improved cardiac function and increased survival in rats with septic myocardial injury. Cellular experiments showed that curcumin increased the survival rate of lipopolysaccharide-treated cardiomyocytes and down regulated TLR1 expression and inhibited NF-κB phosphorylation in cells in a dose-dependent manner. Molecular docking analysis revealed that curcumin interacted with TLR1 by hydrogen bonding and could be stably bound to inhibit the biological function of TLR1. ConclusionOur study shows that curcumin attenuates myocardial injury in sepsis by inhibiting TLR1 expression, which provides a molecular theoretical basis for clinical treatment.

Influence of dietary Bacillus coagulans and/or Bacillus licheniformis-based probiotics on performance, gut health, gene expression, and litter quality of broiler chickens

Probiotics are non-pathogenic microorganisms that are potentially important non-antibiotic alternatives. This study aimed to compare novel multi-strain and single-strain Bacillus probiotics and their respective influences on broiler chickens’ performance, gut health, litter quality, immune response, and NBN and TLR gene expression. A total of 1200 Arbor-Acres 1-day-old broiler chicks were randomly allocated into three treatments (T1 was a control, T2 was supplemented with a combined Bacillus coagulans (2 × 109 cfu/g) and Bacillus licheniformis (8 × 109 cfu/g) probiotic strains (0.2 kg/ton of feed), and T3 was supplemented with Bacillus licheniformis (3.2 × 109 cfu/g) probiotic (0.5 kg/ton of feed) with eight replicas of each. Supplementing the broiler diet with either the single-strain (T3) or the multi-strain (T2) Bacillus-based probiotic raised the overall birds’ body weight, body weight gain, feed conversion ratio, and European production efficiency factor compared to the control (T1), with a significant enhancement achieved by the multi-strain Bacillus product (P = 0.005). T2 and T3 exhibited significantly improved cholesterol, Alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and alkaline phosphatase levels than the control (P ≤ 0.05). The transcript levels of both NBN and TLR genes were upregulated in the liver in the T2 and T3 groups. The T2 group experienced significant reductions in gut bacterial counts, especially for Clostridia, and recorded the lowest litter moisture and nitrogen. In conclusion, supplementing broiler diets with probiotics of multiple Bacillus strains increased production profitability by promoting bird growth, improving feed intake, enhancing gut mucosa and immune organs, and upregulating genes responsible for immunity. All these inhibit the overgrowth of enteric pathogens and sustain litter quality.

Comprehensive analysis of immune parameters, mRNA and miRNA profiles, and immune genes expression in the gill of rainbow trout infected with infectious hematopoietic necrosis virus (IHNV)

Rainbow trout (Oncorhynchus mykiss) is a species of cold-water fish with important economic values, widely cultivated worldwide. However, the outbreak of infectious hematopoietic necrosis virus (IHNV) caused the large-scale death of rainbow trout and seriously restricted the development of the trout farming industry. In this study, the changes of immune parameters in different periods (6-, 12-, 24-, 48-, 72-, 96-, 120-, and 144 h post-infection (hpi)), transcriptome profiles of 48 hpi (T48G) compared to control (C48G), and key immune-related genes expression patterns were measured in rainbow trout gill following IHNV challenge through biochemical methods, RNA sequencing (RNA-seq), and quantitative real-time polymerase chain reaction (qRT-PCR). The results showed that alkaline phosphatase (AKP), acid phosphatase (ACP), total superoxide dismutase (T-SOD), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities, as well as lysozyme (LZM) and malonaldehyde (MDA) content decreased and then increased during infection, and remained at a high level after 48 hpi (P < 0.05), whereas catalase (CAT) activity showed a significant peak at 48 hpi (P < 0.05). The mRNA and miRNA analysis identified 4343 differentially expressed genes (DEGs) and 11 differentially expressed miRNAs (DEMs), and numerous immune-related DEGs involved in the Toll-like receptor signaling pathway, apoptosis, DNA replication, p53 signaling, RIG-I-like receptor signaling pathway, and NOD-like receptor signaling pathway and expression were significantly up-regulated in T48Gm group, including tlr3, tlr7, tlr8, traf3, ifih1, trim25, dhx58, ddh58, hsp90a.1, nlrc3, nlrc5, socs3, irf3, irf7, casp7, mx1, and vig2. The integrated analysis identified several important miRNAs (ola-miR-27d-3p_R+5, gmo-miR-124-3-5p, ssa-miR-301a-5p_L+2, and ssa-miR-146d-3p) that targeted key immune-related DEGs. Expression analysis showed that tlr3, tlr7, traf3, ifih1, dhx58, hap90a.1, irf3, irf7, and mx1 genes increased and then decreased during infection, and peaked at 72 hpi (P < 0.05). However, trim25 expression peaked at 96 hpi (P < 0.05). This study contributes to understanding immune response of rainbow trout against IHNV infection, and provides new insights into the immune regulation mechanisms and disease resistance breeding studies.

TLR2 Gene Polymorphism and MicroRNA 301 gene Expression Level as a Signature for Hepatocellular Carcinoma in Egyptian Patients

TLR2 Gene Polymorphism and MicroRNA 301 gene Expression Level as a Signature for Hepatocellular Carcinoma in Egyptian Patients

Forecasting most deleterious nsSNPs in human TLR9 gene and their cumulative impact on biophysical features of the protein using in silico approaches

In women, the uterine cervix and corpus uteri are two main suspects, playing a major role in cancer-associated-mortality. Immunologically, Toll-like receptors (TLRs) associated with the innate immune system, can recognize pathogens and induce immune responses against pathogens. Cellularly, TLR9 expression occurs in immune system cells including macrophages, natural killer cells, dendritic cells, and other antigen-presenting cells. TLR9 recognizes and interacts with viral and bacterial DNA comprising cytosine-phosphate-guanine (CpG) dideoxynucleotide motif. The current study is designed to identify the most deleterious nonsynonymous single nucleotide polymorphisms (nsSNPs) in the TLR9 gene and to delineate their deleterious effect on the structural and functional features of proteins at the molecular level. Based on the implementation of various computational tools and algorithms eight most deleterious nsSNPs (P139H, R257C, C265Y, L283P, G514D, L544Q, H566Y, and W670R) have been identified in the human TLR9 gene as potentially damaging SNPs. Further, our study suggests highly conserved patterns at deleterious nsSNPs sites could influence protein stability and its functional features. Additionally, this study identifies two nsSNPs (G514D and W670R) associated with the severity of Uterine corpus endometrial carcinoma. In support of our computational findings, the validation of key results using polymerase chain reaction and other experimental methods is warranted in the Indian population. In general, this study might be able to delineate the guideline for identifying the most damaging SNPs and enhances the understating of the risk factors for cancer and disease susceptibilities.

Inflammation Regulation by Bacterial Molecular Patterns.

Stimulation of innate immunity by bacterial molecular patterns can induce an enhanced cellular immune response to pathogens that are associated with innate immune memory shaped by epigenetic changes. Immunological memory can be expressed in the acceleration/intensification of inflammation, as well as in the exact opposite-to maintain tolerance and non-response to a repeated stimulus. Tolerance is one of the central concepts of immunity and is ensured by the consistency of all parts of the immune response. The severe consequences of inflammation force researchers to study in detail all stages of the downstream pathways that are activated after exposure to a stimulus, while the formation of non-response to a pro-inflammatory stimulus has not yet received a detailed description. Elucidation of the mechanism of tolerance is an urgent task for the prevention and treatment of inflammatory diseases. The aim of this investigation was to study the dynamic changes in the gene expression of A20 and ATF3, the inflammation suppressors, against the background of the expression of the genes of the innate immunity receptors TLR4 and NOD2 and the pro-inflammatory cytokine TNF-α under the influence of TLR4 and NOD2 agonists, lipopolysaccharide (LPS) and glucosaminylmuramyl dipeptide (GMDP). The mechanism of inflammation regulation by bioregulators of bacterial origin-LPS and GMDP-was evaluated in vitro in human peripheral blood mononuclear cells and in vivo after i.p. administration of LPS and GMDP to mice. Gene expression was assessed by RT-PCR. Innate immune receptors and the pro-inflammatory cytokine TNF-α were found to develop early in response to LPS and GMDP, both in vitro and in vivo. Genes of cytosolic proteins controlling inflammation (A20 and ATF3) were expressed later. Prior exposure of the innate immune system to LPS and muramyl peptides may modulate host defense against acute inflammation. As a result of the study, new data were obtained on dynamic changes in deubiquitinase A20 and the transcription factor ATF3, which are involved in the limitation and suppression of inflammatory reactions caused by fragments of bacterial cell walls-LPS and GMDP. Thus, bioregulators of bacterial origin LPS and GMDP, along with pro-inflammatory factors, activate the expression of genes that suppress inflammation, which should be considered when analyzing data from studies of the pro-inflammatory properties of LPS and GMDP and when developing drugs based on them.

The effects of dietary supplementation with mushroom or selenium enriched mushroom powders on the growth performance and intestinal health of post-weaned pigs

BackgroundThere is an urgent need to identify natural bioactive compounds that can enhance gastrointestinal health and promote pig growth performance in the absence of pharmacological levels of zinc oxide (ZnO). The objectives of this study were to: 1) compare the effects of mushroom powder supplemented with inorganic selenium (inSeMP) to mushroom powder enriched with organic selenium (orgSeMP) to pharmacological levels of ZnO on growth performance and faecal scores (FS) for the first 21 d post-weaning (Period 1); and 2) compare the molecular and microbial effects of inSeMP and orgSeMP in these pigs on d 39 post-weaning (Period 2).MethodsIn Period 1, pigs (3 pigs/pen; 8 pens/treatment) were assigned to: (1) basal diet (control); (2) basal diet + zinc oxide (ZnO) (3100 mg/kg d 1–14, 1550 mg/kg d 15–21); (3) basal diet + mushroom powder supplemented with inorganic selenium (inSeMP) containing selenium (selenite) content of 0.3 mg/kg feed; (4) basal diet + mushroom powder enriched with organic selenium (orgSeMP) containing selenium (selenocysteine) content of 0.3 mg/kg feed. Mushroom powders were included at 6.5 g/kg of feed.ResultsIn Period 1, there was no effect of diets on average daily gain (ADG) and gain:feed (G:F) ratio (P > 0.05). The orgSeMP supplemented pigs had a lower average daily feed intake (ADFI) compared to all other groups (P < 0.05). The ZnO supplemented pigs had reduced FS compared to the basal and mushroom group, while the orgSeMP supplemented pigs had lower FS compared to the basal group during the 21 d experimental period (P < 0.05). In Period 2, there was no effect of diets on ADFI, ADG and G:F ratio (P > 0.05). The orgSeMP supplementation increased the caecal abundance of bacterial members of the Firmicutes and Bacteroidetes phylum, including Lactobacillus, Agathobacter, Roseburia, and Prevotella and decreased the abundance of Sporobacter compared to the basal group, while inSeMP increased the caecal abundance of Prevotella and decreased the caecal abundance of Sporobacter compared to the basal group (P < 0.05). Dietary supplementation with inSeMP increased expression of TLR4 and anti-inflammatory cytokine gene IL10 and decreased nutrient transporter gene FABP2 compared to the orgSeMP group (P < 0.05).ConclusionOrgSeMP is a novel and sustainable way to incorporate selenium and β-glucans into the diet of weaned pigs whilst improving FS and modulating the caecal microbiota.

Molecular Genetics of Thrombotic Myeloproliferative Neoplasms: Implications in Precision Oncology.

Classical BCR-ABL-negative myeloproliferative neoplasms (MPN) include polycythaemia vera, essential thrombocythaemia, and primary myelofibrosis. Unlike monogenic disorders, a more complicated series of genetic mutations are believed to be responsible for MPN with various degrees of thromboembolic and bleeding complications. Thrombosis is one of the early manifestations in patients with MPN. To date, the driver genes responsible for MPN include JAK2, CALR, MPL, TET2, ASXL1, and MTHFR. Affords have been done to elucidate these mutations and the incidence of thromboembolic events. Several lines of evidence indicate that mutations in JAK2, MPL, TET2 and ASXL1 gene and polymorphisms in several clotting factors (GPIa, GPIIa, and GPIIIa) are associated with the occurrence and prevalence of thrombosis in MPN patients. Some polymorphisms within XRCC1, FBG, F2, F5, F7, F12, MMP9, HPA5, MTHFR, SDF-1, FAS, FASL, TERT, ACE, and TLR4 genes may also play a role in MPN manifestation. This review aims to provide an insightful overview on the genetic perspective of thrombotic complications in patients with MPN.

Physical Activity Maintain Immune Response Through TLR-2/TLR-4 Gene Expression in Type-2 Diabetes Mellitus Patient at Medan City.

The Increasing in type-2 diabetes mellitus (T2DM) needs to solve comprehensively and holistically. Patients with T2DM should have self-coping due to lifestyle modification. Abdominal fat accumulation can release pro-inflammatory cytokine that leads TLR-2 and TLR-4 to the response. These two kinds of toll-like receptors exist on the monocyte surface membrane which is an innate immunity cell. The aims of this study were to get the profile of physical activity, metabolic state, and mononuclear cell response to the expression of the TLR2 and TLR4 genes in T2DM patients. It was a descriptive-analytic study with a cross-sectional study design. Thirty-two eligible patients with inclusion criteria participated as subjects. All subjects answered questions by IPAQ, and checked metabolic state with body composition analysis. The TLR2 and TLR4 gene expression was determined with quantitative Real- Time PCR. This study result found that most T2DM patients were in a highly active category in which most of their activity was walking (light intensity). The average abdominal circumferences were 91.81 ± 15.4 cm, body fat percentage was 29.5 ± 8.8%, and fasting blood sugar was 187.07 ± 67.03 mg/dl. Mononuclear cells number were normal. The expression of the TLR2 gene was lower by 0.71 fold and TLR4 gene expression was lower by 0.9 fold compared with non-DM (p<0.05). By chi-square test, there was a positive correlation between TLR2 gene expression with fasting blood glucose (p=0.011, and a positive correlation between the abdominal circumference and TLR4 gene expression (p=0.011). Type-2 Diabetes mellitus patients in primary health care keep walking as their physical activity to maintain blood glucose. Patients need to do moderate to vigorous exercise regularly to reduce body fat percentage especially abdominal fat to reduce Toll-like receptor gene expression, so insulin resistance and blood glucose level might decline to normal.

Polymorphism study of TLR4 gene in crossbred cattle of Kerala

The present study was conducted to identify single nucleotide variations in exon 3 region of the bovine Toll-Like Receptor gene (TLR4) associated with somatic cell score (SCS) in crossbred cattle of Kerala using polymerase chain reaction-single stranded conformational polymorphisms (PCR-SSCP) technique. Blood samples were collected from 200 lactating crossbred cattle in different farms of Kerala Veterinary and Animal Sciences University (KVASU) and field centres of ICAR- FPT scheme, KVASU. Genomic DNA was isolated and polymorphisms were identified by SSCP analysis. The 231 bp fragment of exon 3 in TLR4 gene was found to polymorphic and two genotypes viz. CC and CD were obtained with frequencies 77 per cent and 23 per cent respectively. The frequencies of C and D allele in crossbred were found to be 0.88 and 0.12. On sequencing, C to T transition had led to one non-synonymous change at 2021th position of ORF. Cattle with CC genotype were associated with significantly lower SCS (p≤0.05) as compared to CD genotype. The association between TLR4 polymorphism and SCS described in this study suggests the possible role of SNP during mastitis infection. Thus, could be employed as a useful marker for genetic selection to promote mastitis resistance in dairy cattle.

Gene Expression of Immune, Growth and Lipogenic Markers in Holstein, Simmental and Brown Swiss Calves

The objective of this study was to explore mRNA levels of immune, growth and lipogenic markers in Holstein Fresian, Simmental and Brown Swiss calves. For this purpose, a total of 180 healthy male calves of three cattle breeds (Holstein Friesian, Simmental and Brown Swiss) with 60 animals from each breed were integrated in this study. Blood samples were collected from jugular vein from each individual calf using vacutainer needle into tubes containing disodium EDTA as an anticoagulant for RNA extraction. There was a significant difference between the three breeds in birth weights of calves, weaning weight of calves, body weights at both 4 months of age and body weight at 6 months of age with the Simmental breed showing the highest value for each parameter. The expression pattern of TLR2, TLR4, TLR6 and BDEF5 genes was significantly up-regulated in Brown Swiss calves compared to Holstein Friesian and Simmental (P&lt;0.05) as evidenced by the diminished levels of ΔCT. However, levels of IGF-1, LPL, SCD and ACACA were significantly up-regulated in Simmental compared to Holstein Fresian and Brown Swiss calves as proved by lowered values of ΔCT in Simmental than Holstein Fresian and Brown Swiss. The results herein indicated that gene expression profile of investigated markers could be used as biomarkers for rigorous selection for immune and growth traits in cattle.

Nanoemulsion of the Essential Oil from Blumea balsamifera (L.) DC. and Its Effect on Trauma Repair.

The essential oil, extracted from the Hmong medicine Blumea balsamifera (L.) DC. (BBO), is a purely natural wound repair agent. Its application has, however, been restricted due to its low solubility and high volatility properties. In this study, we have developed a nanoemulsion formulation to improve the characteristics of BBO. The particle size of the nanoemulsion was normally distributed, and 71% of its range was concentrated between 10-100 nm, with an average particle size of 62.8 nm and an encapsulation rate of 98%. After 7 days of application, the wound healing rate of the BBO nanoemulsion (BBO-NE) group was 1.5 times higher than that of the normal BBO group. Along with histological observations, nanoemulsion formulation has been demonstrated to significantly improve the efficacy of BBO for wound repair. In addition, inflammation-related TLR4, CD14 and IRAK-1 gene transcript levels were significantly reduced after the administration of BBO-NE compared to the BBO group, with downregulation of 47.8%, 35.7% and 57.8%, respectively, while the secretion of pro-inflammatory factors IL-6 and TNF-α was also significantly reduced by 83.8% and 32.7%, respectively, in the nanoformulation administration (BBO-NE) group compared to the BBO group. In contrast, the anti-inflammatory factor IL-10 was significantly increased by 4.2-fold. It was further found that the drug penetration per unit area increased significantly 6.30% to 19.5% at different time points after the application of the BBO-NE compared to the BBO. In conclusion, nano-formulation enhanced the drug penetration of the BBO, reduced inflammatory factors, increased the level of anti-inflammatory factors, and promoted collagen deposition, thereby accelerating wound repair.

Relationship of Toll-Like Receptors 2 and 4 Gene Polymorphisms with Essential Hypertension in Chinese Han Population

Objective: There are numerous studies suggesting that genetic polymor-phisms of inflammation factors Toll-like receptors 2 and 4 (TLR2, TLR4) might play a role in the pathophysiological process of hypertension. In this study, we evaluated the association in a sample of members of the Chinese Han population. Method: We selected four single nucleotide polymor-phisms (SNP) of TLR2 (rs3804099, rs3804100, rs7656411) and TLR4 (rs1927906) genes, and measured the distributions of genotypic and allelic frequencies in 1063 participants, including 391 essential hypertension pa-tients and 672 controls. Result: No significant differences in the genotypic and allelic frequencies of the four SNPs were detected between cases and controls. However, three haplotypes, CCG, TTG and TTT of TLR2, were significantly associated with a decrease in the risk of essential hyperten-sion (OR: 0.512, 95% CI: 0.397 - 0.660, P P = 0.0038; OR: 0.797, 95% CI: 0.667 - 0.952, P = 0.0122, respectively). Inversely, the risk of essential hypertension increased sig-nificantly in patients with the CTG, TCG or TCT haplotypes (OR: 2.924, 95% CI: 2.157 - 3.963, P P P TLR2 might have profound effects on the development of essential hypertension in the Chinese Han population.

Association between 15 insertion/deletion genetic polymorphisms and risk of schizophrenia using pooled samples.

Schizophrenia is a psychiatric syndrome that affects approximately 1 % of the world population and is among the top 10 reasons for disability. In this case-control study, we investigated the association between 15 insertion/deletion (Indel) polymorphisms and schizophrenia risk using pooled samples. In the present case-control study, 361 individuals with schizophrenia and 360 healthy individuals were included in the study. We examined the insertion/deletion polymorphisms in APOB, ADRA2B, PDCD6IP, LRPAP1, TLR2, DHFR, VEGF, HLA-G, TPA, DBH, UCP2, FADS2, MDM2, TP53 and SLC6A4 genes. Our results revealed that the Del allele of the HLA-G 14bp Indel polymorphism increased the risk of schizophrenia (OR=1.23, 95 % CI=1.01-1.52, p=0.045) and the Alu- allele of the TPA Alu+/Alu- polymorphism negatively associated with the schizophrenia risk (OR=0.67, 95 % CI=0.54-0.82, p<0.001).