Tumor Necrosis Factor-alpha Gene Research Articles

Association of Single Nucleotide Polymorphism in the Upstream Region of Tumor Necrosis Factor Alpha Gene with Asthma

Association of Single Nucleotide Polymorphism in the Upstream Region of Tumor Necrosis Factor Alpha Gene with Asthma

Analysis of Association between Interleukin-6 (<i>IL</i>6), Interleukin-13 (<i>IL</i>13) and Tumor Necrosis Factor-Alpha (<i>TNF-alpha</i>) Gene Polymorphisms and Genetic Susceptibility of Rheumatoid Arthritis in Kuwaiti Arab

Background: Rheumatoid arthritis (RA) is a common autoimmune disease in which a combination of risk alleles from different susceptibility genes predisposes the patients to develop clinical symptoms following exposure to environmental factors. RA is a chronic and progressive disease characterized by synovial inflammation that results in destruction in the affected joints and severe problems in individual’s mobility. Several immune-related risk factors have been associated with RA, these include single nucleotide polymorphisms (SNPs) in cytokine genes. The impact of these cytokine gene polymorphisms is due to their association either with elevated serum levels and/or variations in their serum levels are associated with disease-onset and progression. The objective of this study was to investigate the role of Interleukin-6 (IL6), Interleukin-13 (IL13) and Tumor necrosis factor-alpha (TNF-alpha) gene polymorphisms in genetic susceptibility of RA in Kuwaiti patients. Methods: We have determined the genotypes of IL6 gene (-174G/C; rs1800795), IL13 gene (R130Q; rs20541) and TNF-alpha gene (-308A/G' rs1800629) polymorphisms in 192 Kuwaiti patients with RA and compared it to that in 104 healthy controls. The diagnosis of RA was based on the American College of Rheumatology (ACR) classification criteria. The genotypes for IL6, IL13 and TNF-alpha gene polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods and confirmed by DNA sequencing. Results: The frequency of IL6 gene (-174G/C; rs1800795) and TNF-alpha gene (-308A/G' rs1800629) polymorphisms manifested a statistically significant difference between Kuwaiti RA patients and controls (P = 0.02 and 0.002 respectively). In contrast, the frequency of IL13 gene (R130Q; rs20541) polymorphism did not show a significant difference between Kuwaiti RA patients and controls. Conclusions: Our data showed an association of two cytokine gene polymorphisms (i.e. IL6 gene -174G/C; rs1800795 polymorphism, and TNF-alpha gene -308A/G, rs1800629 polymorphism) with RA in Kuwaiti patients highlighting their significant contribution in genetic susceptibility of this chronic disease possibly along with other factors.

Dietary Chlorella vulgaris modulates the performance, antioxidant capacity, innate immunity, and disease resistance capability of Nile tilapia fingerlings fed on plant-based diets

This study assessed the impacts of dietary Chlorella vulgaris (ChV) on the growth, digestive enzymes activity, antioxidant status, immune responses, expression of hepatic cytokines and growth-related genes, and disease resistance capability of Nile tilapia (Oreochromis niloticus) fingerlings fed on plant-based diets. For this purpose, six tested diets were formulated to include 0.0 (control), 2.5, 5, 10, 15, and 20 g ChV/kg diet and were offered to the fish in triplicated groups for 70 days. A dose-dependent increase was observed in feed intake, specific growth rate, and weight gain (%) in ChV-fed fish but feed conversion ratios and survival rates % were not significantly changed among different ChV groups. Significantly higher serum protease, amylase, and lipase enzyme levels were detected in ChV-fed fish than the control group, especially in treatments of 10 – 15 g/kg diet. Feeding the fish with 10–20 g ChV/kg diet markedly increased serum total proteins, total lipids, and globulin levels. Similarly, glutathione peroxidase, superoxide dismutase, and catalase activities as well as serum lysozyme and respiratory burst activities, and total IgM levels were also elevated. On the other hand, lowest malondialdehyde levels were recorded in fish groups fed diets with 10 – 20 g ChV/kg diet compared with the control group. Bactericidal activities against Aeromonas hydrophila, Pseudomonas fluorescens, and Vibrio anguillarum were significantly higher as levels of dietary ChV increased up to 15 and 20 g/kg diet. Dietary ChV also upregulated the transcription of hepatic growth hormone, insulin-like growth factor 1, interleukin-1 beta, and tumor necrosis factor-alpha genes, particularly at 15 – 20 g/kg diet. However, feeding Nile tilapia fingerlings with 15 g ChV/kg diet is adequate to promote the performance and support antioxidant capacity, immune response, and resistance capability to bacterial infections.

Contribution of -1031T/C and -376G/A tumor necrosis factor alpha polymorphisms and haplotypes to preeclampsia risk in Tunisia (North Africa)

Preeclampsia is a gestational disorder characterized by hypertension and proteinuria. Excessive release of pro-inflammatory cytokines, particularly tumour necrosis factor-alpha, has been demonstrated to contribute to endothelial activation and poor trophoblast invasion in placental development, resulting in preeclampsia's clinical symptoms. Genetic polymorphisms of tumour necrosis factor-alpha can regulate its production and may play an important role in the pathogenesis of this disease. This study aimed to evaluate the association of five tumour necrosis factor-alpha gene promoter single nucleotide polymorphisms, or their haplotype combinations, with preeclampsia prevalence. This case-control study was conducted on 300 women with preeclampsia and 300 age-matched women with normal pregnancy from Tunisian hospitals. Genotyping of tumour necrosis factor-alpha −1031 T/C, −376 G/A, −308 G/A, −238 G/A, and +489 G/A SNPs was performed on DNA extracted from blood samples using PCR-restriction fragment-length polymorphism analysis. Statistical analysis was performed using the chi-square test. P < 0.01 were considered statistically significant to take into consideration the multiple comparisons. A significantly higher frequency of the minor allele -1031C (p < 0.001) was observed in preeclampsia cases compared to controls. Notably, the -1031C and -376A (CA) haplotype, which correlates with a higher production of TNF-α protein, had a higher incidence in women with preeclampsia (p = 0.0005). Conversely, the TG haplotype had a low frequency in preeclampsia cases compared to controls (p = 0.002) which suggests that it is associated with a reduced incidence of preeclampsia. These results suggest that tumour necrosis factor-alpha polymorphisms, in particular the -1031C/A, and the haplotype CA, contribute to an increased risk of preeclampsia in Tunisian women.

Fig (Ficus exasperata and Ficus asperifolia)-Supplemented diet improves sexual function, endothelial nitric oxide synthase and suppresses tumour necrosis factor-alpha genes in hypertensive rats.

We measured the effect of varieties of Fig leaves, Ficus exasperata Vahl. (FE) and Ficus asperifolia Miq (FA), commonly found in Sub-sahara Africa for managing hypertension on sexual performance in hypertensive rats, which is unknown. Hypertensive rats experienced erectogenic damage after exposure to 40mgkg-1 bw-1 , N(G)-nitro-l-arginine-methyl-ester (L-NAME). Experimental rats were grouped into eight groups (n=6) namely: control rats, hypertensive rats, hypertensive treatment groups with atenolol (10mgkg-1 day-1 ) and sildenafil (5.0mg/kg), rats treated with FE- and FA-formulated biscuits at proportions of 2.5g and 5.0g respectively. Furthermore, we measured the level of sexual performance (Intromission number, latency, Mounting number, and latency), hormonal levels, phosphodiesterase-5 enzyme (PDE-5) activity and genes expressed using real-time quantitative polymerase chain reaction (RT-qPCR) in hypertensive rats. The result deduced revealed that treated hypertensive rats showed significantly reduced follicle-stimulating hormone, luteinising hormone levels, PDE-5 enzyme activity, tumour necrosis factor-alpha (TNF-α) expression while having a marked increase in testosterone level, sexual performance and endothelial nitric oxide synthase (eNOS) expression. Therefore, this study revealed the comparative dietary enhancing effect of FE- and FA-formulated biscuit on sexual behaviour activity, hormonal levels and the level of eNOS and TNF-α genes expressed in hypertensive rats.

Investigation of the transcriptomic response in Atlantic salmon (Salmo salar) gill exposed to Paramoeba perurans during early onset of disease

Amoebic Gill Disease (AGD), caused by the protozoan extracellular parasite Paramoeba perurans (P. perurans) is a disease affecting Atlantic salmon (Salmo salar). This study investigated the gill transcriptomic profile of pre-clinical AGD using RNA-sequencing (RNA-seq) technology. RNA-seq libraries generated at 0, 4, 7, 14 and 16 days post infection (dpi) identified 19,251 differentially expressed genes (DEGs) of which 56.2% were up-regulated. DEGs mapped to 224 Gene Ontology (GO) terms including 140 biological processes (BP), 45 cellular components (CC), and 39 molecular functions (MF). A total of 27 reference pathways in the Kyoto Encyclopedia of Genes and Genomes (KEGG) and 15 Reactome gene sets were identified. The RNA-seq data was validated using real-time, quantitative PCR (qPCR). A host immune response though the activation of complement and the acute phase genes was evident at 7 dpi, with a concurrent immune suppression involving cytokine signalling, notably in interleukins, interferon regulatory factors and tumour necrosis factor-alpha (tnf-α) genes. Down-regulated gene expression with involvement in receptor signalling pathways (NOD-like, Toll-like and RIG-1) were also identified. The results of this study support the theory that P. perurans can evade immune surveillance during the initial stages of gill colonisation through interference of signal transduction pathways.

Association of tumor necrosis factor-α (−308 G/A) and interleukin-10 (−1082 A/G) gene polymorphisms with polycystic ovary syndrome in Iraqi women

BackgroundThe etiologic factors of polycystic ovary syndrome (PCOS) are not well understood, however, several studies reported that genetics and environmental factors might be involved in the appearance of PCOS. Therefore understanding some molecular pathways can provide more information about PCOS. The imbalance of pro-inflammatory and anti-inflammatory cytokines could be associated with its pathophysiology. There are huge numbers of polymorphisms within cytokine genes that may affect their production and bring in the women more susceptible for PCOS. The aim of the present study was for evaluation the correlation of single nucleotide polymorphism (−308 G/A) in tumor necrosis factor-alpha (TNF-α) gene and (−1082 A/G) in interleukin-10 (IL-10) gene with PCOS. MethodsPatients group included 80 women who had a history of PCOS diagnosed using transvaginal or transabdominal ultrasound and don't have any of the inherited disorders such as androgen-secreting neoplasms, congenital adrenal hyperplasia, thyroid dysfunction, and hyperprolactinemia with a mean age (27.23 year). The control group included 70 women with a mean age (26.89 year). Levels of serum IL-10 and TNF-α in woman with PCOS and healthy control have been measured by Enzyme linked immunosorbent assay (ELISA). Single nucleotide polymorphism (SNP) in TNF-α (−308 G/A) and IL-10 (−1082 A/G) genes and the frequency of mutations in patient and control group were evaluated using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). ResultsShowed that serum IL-10 concentration was significantly decreased (p < 0.01) while TNF-α level was significantly increased (p < 0.01) in patients compared to control group. While the genotype prevalence of TNF-α (−308 G/A) and IL-10 (−1082 A/G) and the allele frequency show non-significant difference (p > 0.05) between women with PCOS and healthy controls. ConclusionIt is concluded that the level of TNF–α and IL-10 could be considered as possible biomarkers for PCOS while polymorphisms in IL-10 gene and TNF-α gene are not considered as risk factors of PCOS in Iraqi women, this suggests that further studies on other loci or other cytokines are required.

Experimental models of orthodontic tooth movement and their effects on periodontal tissues remodelling

ObjectivesThe present study aimed to compare two different models of orthodontic tooth movement (OTM) in rats by evaluating tooth movement efficiency and periodontal tissues remodelling. DesignFifteen animals were randomly distributed into 3 groups: control group (untreated); ligature appliance (LA) as experimental OTM using a closed coil spring fixed around maxillary first molar by steel ligature; occlusal appliance (OA) as experimental OTM using a closed coil spring attached on the occlusal surface of the maxillary first molar. After 15 days, all animals were euthanized, and the maxilla of each animal was collected for qPCR, micro-computed tomography, and histological analyses. ResultsInterleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha gene expressions were significantly upregulated in the animals of the LA group as compared to the other groups. No significant difference was observed in tooth displacement between both methods. The LA group presented higher linear bone loss and lower values of bone volume fraction, bone mineral density, trabecular number and increased values of trabecular separation compared to the other groups. The birefringent collagen content in the tension side of the periodontal ligament contained significantly lower collagen content in the LA group than in the control group. Furthermore, on the pressure side, the collagen content was significantly lower in the LA and OA groups than in the control group. ConclusionsThe OA group presented little or no deleterious effect on periodontal tissues compared to the LA group, suggesting its use may be more reliable for OTM induction in rats for 15 days.

The Use of High Resolution Melting (HRM) Method to Detect rs1800629 of Tumor Necrosis Factor-alpha (TNF-alpha) Gene among Tuberculosis Patients

The rs1800629 polymorphism plays a crucial role in the pathogenesis of infectious and autoimmune diseases. Meanwhile, tuberculosis (TB) remains a health primary infectious disease in Indonesia. The purpose of this study is to evaluate the HRM method in detecting the rs1800629 genotype, in the TNF-α gene’s promoter region, within TB patients. The benefit of this study is to accelerate the detection of rs1800629 with a simple, rapid, and cost-effective method for genotyping and mutation screening that does not include the use of a fluorescent probe. In this experimental study, the rs1800629 genotyped in a total of 25 tuberculosis patients using KAPA HRM kit in MyGo Mini PCR, and all amplified PCR products subsequently dispatched for direct DNA sequencing to Macrogen Inc, South Korea. Based on the results, a 100% concordance find in the genotyping of rs1800629 between HRM and sequencing. The authors provided evidence to use HRM in detecting rs1800629 within the TNF-α promotor region. This application as a genotyping assay in tuberculosis patients is a low-cost, rapid, and accurate detection. However, further studies using the HRM method in case-control samples of tuberculosis are required to evaluate the method’s effectiveness and to obtain more information regarding the genotype’s susceptibility to tuberculosis and its adverse effect treatment, including anti-tuberculosis drug, induced liver injury (AT-DILI), and multidrug-resistant TB (MDR-TB), within the Indonesian population.

Ginkgo biloba leaf extract improves growth, intestinal histomorphometry, immunity, antioxidant status and modulates transcription of cytokine genes in hapa-reared Oreochromis niloticus

This study was designed to evaluate the impacts of dietary supplementation with Ginkgo biloba leaf extract (GBL) on the growth, intestinal histomorphometry, immunity, antioxidant status, and expression of cytokine genes in Nile tilapia reared in the hapas. A control diet was enriched with different GBL levels (0.0, 5.0, 7.0, and 9.0 g/kg) to form 4 experimental diets and were fed to Nile tilapia for 8 weeks. The findings illustrated that dietary GBL significantly enhanced the growth and feed utilization indices compared to those reared in the control group. A dose-dependent increase of hepatic catalase, superoxide dismutase, and glutathione peroxidase activities alongside a decline of hepatic malondialdehyde levels were recorded in GBL groups compared with the control. Serum lysozyme activity, complement C3, and immunoglobulin M levels were significantly increased in GBL groups compared with the control group. Moreover, dietary GBL maintained the normal intestinal and hepatopancreatic histological structures with a significant increase of some histomorphometric measurements of proximal, middle, and distal intestinal parts of the treated fish. Interestingly, dietary GBL supplementation significantly increased the mRNA expression of interleukin-1 beta (IL-1β), IL-6, IL-10, tumor necrosis factor alpha (TNF-α) and interferon gamma (INF-γ) genes in the splenic tissues of treated fish over the control group. To conclude, it could be recommended to use GBL as a functional phytogenic feed additive to improve the growth, hepatic and intestinal health status, hepatic antioxidant status, and immunity of treated Nile tilapia. Besides, the second order polynomial regression revealed that 7.50 g GBL/kg diet is the optimal inclusion level to improve growth with no negative impacts on the overall health condition of treated Nile tilapia.

Tumor necrosis factor alpha gene level in patients with tympanosclerosis

Background: Tympanosclerosis is the calcification of connective tissue in the middle ear, particularly the tympanic membrane, which can arise following middle ear infection. Inflammatory reactions are induced and maintained by tumour necrosis factor (TNF) in a variety of illnesses, including chronic otitis media with effusion (COME). The goal of this study was to determine the plasma level of Tumer necrosis factor (TNF) gene expression in individuals who had tympanosclerosis as a result of ventilation tube insertion. Methods: This study comprised sixty patients who were separated into two groups: 60 patients were assigned to Group A based on inclusion and exclusion criteria, whereas 20 volunteers were assigned to Group B. Patients were given a medical history, a thorough physical examination, and a TNF- gene expression test in the lab (qRT-PCR). Results: There was a significant difference in TNF-gene expression across groups, with the mean TNF in the cases group (3.87) being significantly greater than the control group (1), p0.001. TNF levels were statistically different depending on the number of tubes inserted (p=0.005) and the time of secretory OM (p=0.042). There was no significant difference in TNF levels when it came to medication usage or acute OM episodes. Conclusion: TNF- gene profile expression was statistically significant higher in selected middle-ear structures with tympanosclerosis than in the control group.

Blastocystis Colonization Alters the Gut Microbiome and, in Some Cases, Promotes Faster Recovery From Induced Colitis.

Protists are a normal component of mammalian intestinal ecosystems that live alongside, and interact with, bacterial microbiota. Blastocystis, one of the most common intestinal eukaryotes, is reported as a pathogen that causes inflammation and disease, though health consequences likely vary depending on host health, the gut ecosystem, and genetic diversity. Accumulating evidence suggests that Blastocystis is by and large commensal. Blastocystis is more common in healthy individuals than those with immune mediated diseases such as Inflammatory Bowel Diseases (IBD). Blastocystis presence is also associated with altered composition and higher richness of the bacterial gut microbiota. It is not clear whether Blastocystis directly promotes a healthy gut and microbiome or is more likely to colonize and persist in a healthy gut environment. We test this hypothesis by measuring the effect of Blastocystis ST3 colonization on the health and microbiota in a rat experimental model of intestinal inflammation using the haptenizing agent dinitrobenzene sulfonic acid (DNBS). We experimentally colonized rats with Blastocystis ST3 obtained from a healthy, asymptomatic human donor and then induced colitis after 3 weeks (short term exposure experiment) or after 13 weeks (long term exposure experiment) and compared these colonized rats to a colitis-only control group. Across experiments Blastocystis ST3 colonization alters microbiome composition, but not richness, and induces only mild gut inflammation but no clinical symptoms. Our results showed no effect of short-term exposure to Blastocystis ST3 on gut inflammation following colitis induction. In contrast, long-term Blastocystis exposure appears to promote a faster recovery from colitis. There was a significant reduction in inflammatory markers, pathology 2 days after colitis induction in the colonized group, and clinical scores also improved in this group. Blastocystis colonization resulted in a significant reduction in tumor necrosis factor alpha (TNFα) and IL-1β relative gene expression, while expression of IFNγ and IL17re/17C were elevated. We obtained similar results in a previous pilot study. We further found that bacterial richness rebounded in rats colonized by Blastocystis ST3. These results suggest that Blastocystis sp. may alter the gut ecosystem in a protective manner and promote faster recovery from disturbance.

DNA methylation of tumour necrosis factor (TNF) alpha gene is associated with specific blood fatty acid levels in a gender-specific manner.

BackgroundFatty acids, specifically polyunsaturated fatty acids (PUFAs) play an important role in inflammation and its resolution, however, their interaction with the epigenome is relatively unexplored. Here we investigate the relationship between circulating blood fatty acids and the DNA methylation of the cytokine encoding gene tumour necrosis factor (TNF, OMIM 191160).MethodsUsing a cross‐sectional study approach, we collected blood samples from adults (N=88 (30 males, 58 females); 18–74 years old) for DNA methylation pyrosequencing analysis at four sites in TNF exon 1 and gas‐chromatography mass‐spectrometry analysis of the fatty acid profile of dried blood spots (DBS).ResultsMethylation levels of TNF exon 1 are significantly correlated with specific fatty acids in a gender‐specific manner. In the males the PUFAs Docosahexaenoic Acid (DHA) and Arachidonic Acid (AA) were positively associated with TNF methylation, as was the saturated fatty acid (SFA) Stearic Acid; in contrast, mono‐unsaturated fatty acids (MUFAs) had a negative association. In the females, omega‐6 PUFA γ‐Linolenic acid (GLA) was negatively correlated with TNF methylation; Adrenic acid and Eicosadienoic Acid were positively correlated with TNF methylation.ConclusionThese results suggest that one way that fatty acids interact with the inflammation is through altered methylation profiles of cytokine genes; thus, providing potential therapeutic targets for nutritional and health interventions.

The Frequency of Tumor Necrosis Factor-alpha Gene Polymorphism in Patients with Chronic Immune Thrombocytopenia.

Background: The aim of this study was to reveal the frequency of Necrosis Factor _Alpha Gene Polymorphism in evaluation of severity and outcome of chronic immune thrombocytopenia in pediatric department of Zagazig University hospitals. Methods: We conduct a retrospective case control on 40 children diagnosed with chronic ITP and 40 age and sex healthy matched subjects as a control recruited from outpatient hematology clinic Faculty of Medicine, Zagazig University. Results: We found a highly significant difference between ITP group and control group as regarding TNF-α alleles where A allele was expressed in 43.7% of cases versus 17.5% 0f control, P value

Bacteriophage‐mediated therapy of chondrosarcoma by selective delivery of the tumor necrosis factor alpha (TNFα) gene

Chondrosarcoma is a cartilage-forming bone tumor, well known for intrinsic resistance to chemotherapy and radiotherapy. We have designed a targeted chondrosarcoma gene therapy using a bacteriophage (phage) particle to deliver therapeutic genes. Phage has no tropism for mammalian cells, allowing engineered phage to be targeted to specific cell surface receptors in cancer. We modified the phage capsid to display the RGD4C ligand on the pIII minor coat proteins to specifically bind to αvβ3 or αvβ5 integrin receptors. The endosomal escape peptide, H5WYG, was also displayed on recombinant pVIII major coat proteins to enhance gene delivery. Finally, a human tumor necrosis factor alpha (TNFα) therapeutic transgene expression cassette was incorporated into the phage genome. First, we found that human chondrosarcoma cells (SW1353) have high expression of αvβ3, αvβ5 integrin receptors, and both TNFα receptors. Targeted particle encoding a luciferase reporter gene efficiently and selectively mediated gene delivery to these cells. When SW1353 cells were treated with the targeted particle encoding a TNFα transgene, significant cell killing was evident and was associated with high expression of TNFα and apoptosis-related genes. In vivo, mice with established human chondrosarcoma showed suppression of tumors upon repetitive intravenous administrations of the targeted phage. These data show that our phage-based particle is a promising, selective, and efficient tool for targeted chondrosarcoma therapy.

DAPAGLIFLOZIN, PIOGLITAZONE AND THEIR COMBINATION EFFECTED LEVELS OF LIPOPROTEINS AND ADIPOKINES IN PLASMA AND THEIR GENE EXPRESSION IN FAT IN RATS WITH DIABETES MELLITUS TYPE 1

Objective: Patients with diabetes mellitus type1 (DM1) have a relative insulin deficiency with elevated triacylglycerides (TAG) production. Adipokines are important in fat distribution, inflammation, and endothelial function. Dapagliflozin (Dapa) and pioglitazone (Pio) are oral hypoglycemic drugs for the treatment of DM2. They have also pleiotropic effects on lipid profile and body weight. The effect of these drugs on lipoproteins and adipokines levels in DM1 is poorly understood. Design and method: DM1 in male Wistar rats was induced by a single dose of streptozotocin (STZ). Animals were then divided into five groups: Controls, diabetic animals (DM), DM treated with Dapa, DM treated with Pio, DM treated with combination DapaPio. Therapies were mixed in rat chow. Control and DM groups received standard chow. After 6 weeks of treatment serum glucose and lipid levels were determined. Visceral adipose tissue (VAT) gene expression of leptin (Lep), adiponectin (AdipoQ), hormone-sensitive lipase (HSL), lipoprotein lipase (LPL), tumor necrosis factor-alpha (TNFa) and interleukin 6 (IL6) were measured by RT qPCR and their levels in plasma were determined by ELISA method. Results: STZ injection led to a significant increase in both fasting and postprandial glucose levels and all levels of lipoproteins except high-density lipoproteins (HDL), which were decreased. We also found decreased levels of Lep, AdipoQ, LPL, and IL6, and increased HSL and TNFa in plasma in DM group. These results were supported by the same significant changes in gene expression in VAT. A combination of DapaPio resulted in a significant decrease in serum levels of both glucose levels. Pio and DapaPio administration decreased TAG and total cholesterol and increased HDL. Lep, AdipoQ, LPL and IL6 plasma levels were increased, and HSL, TNFa were decreased after administration of Dapa, Pio and their combination. Both monotherapy and combination significantly modulated Lep, TNFa and IL6 gene expression in VAT. Conclusions: DM1 in rats influenced not only glucose levels but also lipoproteins and adipokines in plasma, which could be affected by Dapa, Pio and DapaPio. Therapy can also change the gene expression of Lep, AdipoQ, TNFa, and IL6 in adipose tissue in rats with DM1.

Genetic polymorphism of tumor necrosis factor alpha (TNF-α) and tumor necrosis factor beta (TNF-β) genes and risk of oral pre-cancer and cancer in North Indian population.

There are inconclusive data connecting single-nucleotide polymorphisms (SNPs) of TNF-α (rs361525) and TNF-β (rs909253) to potential malignant oral disorder (PMOD) such as lichen planus and oral fibrosis. Here, we have investigated the risk of oral squamous cell carcinoma as well as oral pre-cancerous lesions in North Indian population with the polymorphism of the TNFα/ β genes. A total 500 patients with oral pre-cancer and OSCC and 500 healthy volunteers were genotypes for the TNF-α (-238) G/A (rs361525) and TNF-β (252) A/G (rs909253) gene polymorphism. Genotypes were identified by polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP). Genotype frequencies were evaluated by Chi-square test. Compared to the GG genotype, the GA genotype of TNF-α (G238A) polymorphism (rs361525) has been found to significantly increase the risk of oral disease (OR = 1.99) and especially the risk of lichen planus and OSCC (OR = 2.805 and 5.790, respectively). Similarly, the risk of oral disease was also more in the heterozygote (AG) than the common allele homozygote (AA) of TNF-β (A252G) polymorphism (rs909253) (OR = 1.483). We conclude that the SNPs rs361525 and rs909253 were significantly associated with oral pre-cancer and OSCC.

Comparable in vitro Function of Human Liver-Derived and Adipose Tissue-Derived Mesenchymal Stromal Cells: Implications for Cell-Based Therapy.

Mesenchymal stem/stromal cells (MSCs) have been investigated extensively for their immunotherapeutic and regenerative properties, which may differ by cell source. In MSCs harvested from donors matched for sex, age, and body mass index, we compared the proliferative and migration functions of liver-derived MSCs (L-MSCs) and adipose tissue-derived MSCs (A-MSCs) (n = 6 donors each). Cellular senescence was evaluated by senescence-associated beta-galactosidase enzyme activity and expression of senescence-associated secretory phenotype (SASP) factors using real-time quantitative polymerase chain and by western blot assay. The pro-angiogenic and reparative potency of MSCs was compared by co-culturing MSCs with injured human umbilical vein endothelial cells (HUVEC). The proliferation and migration properties were similar in L-MSCs and A-MSCs. Although cell cycle arrest and SASP genes were similarly expressed in both MSCs, tumor necrosis factor alpha gene and protein expression were significantly downregulated in L-MSCs. In co-cultured injured HUVEC, A-MSCs restored significantly more tubes and tube connections than L-MSCs. Therefore, despite many functional similarities between L-MSCs and A-MSCs, L-MSCs have enhanced immunomodulatory properties, while A-MSCs appear to have better pro-angiogenic and vascular reparative potency. Availability of a broad range of cellular options might enable selecting cell-based therapy appropriate for the specific underlying disease.

Effect of anthocyanin extract from Roselle,Hibiscus sabdariffa, calyx on haematological, biochemical and immunological parameters of rainbow trout,Oncorhynchus mykiss

In the present study, rainbow trout, Oncorhynchus mykiss, (14.1 ± 0.08 g) were fed diets supplemented with different levels of Roselle anthocyanin extract (RE) to assess the fish immune, stress and haematological parameters. For this, four levels of RE including 0, 0.5, 1 and 1.5% were added to the fish diet for eight weeks. Then, growth performance, blood haematological, biochemical, immunological and hepatic expression of cytokine genes were investigated in all treatments. At the end of the experiment, the fish exhibited similar growth rate and survival (final weight: 91.4–96.7 g; feed conversion ratio: 0.75–0.80; specific growth rate: 3.11–3.21% d−1; survival: 100%). The results showed that there were no significant differences in plasma glucose, cholesterol, triglyceride, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatase levels among the treatments. Supplementation of the diets with 1% RE significantly increased RBC, haemoglobin, haematocrit, lysozyme, complement and WBC, compared with the control treatment. 1 and 1.5% RE significantly decreased plasma cortisol levels, compared with the control. All levels of dietary RE significantly up-regulated hepatic tumour necrosis factor-alpha gene expression, compared with the control treatment. 1 and 1.5% RE significantly up-regulated hepatic interleukin-1 beta gene expression, compared with the control treatment. 1.5% RE supplementation significantly down-regulated interleukin-10 gene expression, compared with the other treatments. In conclusion, dietary RE supplementation at 1% is beneficial for rainbow trout, as it mitigates stress and increases immune-related parameters.

Polymorphism of LEP and TNF-α Genes in the Dairy Cattle Populations of Ukrainian Selection

Study was conducted by investigating the genetic structure of cattle populations of Ukrainian Black-and-White and Ukrainian Red-and-White dairy breeds by polymorphism of leptin gene (LEP) and tumor necrosis factor-alpha gene (TNF-α). The studies were carried out using the method of polymerase chain reaction (PCR) and restriction analysis for LEP and using classic PCR with subsequent SSCP analysis in case of TNF-α. According to the results of the study, it was shown that the LEP gene was polymorphic in both experimental populations by HphI-polymorphism in the third exon of the gene (A59V mutation). In the population of the Ukrainian Black-and-White dairy cattle breed, the frequency of allele C (HphI-) was 0.77; allele T (HphI+) – 0.23. Wright's fixation index was 0.23; which indicates a significant excess in the number of homozygous individuals. This population had a deviation from the Hardy-Weinberg equilibrium state. In the population of the Ukrainian Red-and-White dairy breed, the frequency of allele C was 0.72; allele T – 0.28; Wright's fixation index was -0.18. According to the SSCP-polymorphism of the TNF-α gene, 6 alleles with a size of 450-1200 bp were detected (alleles A, B, and F for Ukrainian Black-and-White dairy breed; A, B, C, D, Е, F – for Ukrainian Red-and-White dairy breed). The frequency of allele A prevailed in the populations of both breeds (0.58 and 0.54, respectively). Alleles C, D, and E had a low frequency of occurrence (0.04-0.16) and they were found only in the population of Red-and-White dairy breed.