Tumor Necrosis Factor-alpha Gene Expression Research Articles

Impact of transportation in freshwater and brackish water on Nile tilapia (Oreochromis niloticus) resistance

BackgroundOreochromis niloticus has great economic value and potential for farming and development. Transportation of fish was done for breeding or trading purpose and it is a challenging aspect of aquaculture. This study aimed to investigate the effect of transportation in freshwater and brackish water on the resistance of O. niloticus as well as transportation stress mitigation effect of NaCl. Four equal groups were used; each of 50 fish, the 1st group served as the control (P 1), while the 2nd group (PT 2) was transported in water without salt, the 3rd (PT 3) and 4th (PT 4) groups were transported in water containing 5 gL− 1 and 10 gL− 1 salt respectively. PT 2, PT 3 and PT 4 were transported for 5 h without any rest or sedative drugs.ResultsThe serum cortisol of O. niloticus significantly increased at 0 h and then decreased at 12 and 24 h post transportation in the PT 2 group and non-significantly increased at all point times in the PT 3 and PT 4 groups comparing to P 1 group. Mucin2 gene (MUC2) expression was non-significantly up regulated in the PT 2 group and down regulated in the PT 3 and PT 4 groups at 0 h comparing with P 1 group, but at 12 and 24 h it was significantly up regulated in the PT 2, PT 3 and PT 4 groups. The β Defensin-1 (β D1) and 2 (β D2) genes expression was non-significantly down-regulated in the PT 2 group and significantly up regulated in the PT 3 and PT 4 groups at 0 h., while at 12 and 24 h was significantly down regulated in the PT 2 group and non-significantly down regulated in the PT 3 and PT 4 groups, it significantly down regulated in the PT 2 and PT 3 group and non-significantly down regulated in the PT 4 group at 24 h. Non-significant up regulation in interleukin − 1β (IL-1β) gene expression was reported in the PT 2 group and non-significant down regulation in the PT 3 and PT 4 groups at 0 h. However, significant up regulation was recorded in the PT 2, PT 3 and PT 4 groups at 12 and 24 h. The Tumor necrosis factor-alpha (TNF–α) gene expression was non-significantly up regulated in the PT 2 group and non-significantly down regulated in the PT 3 and PT 4 groups at 0 h. However, it was significantly up regulated in the PT 2, PT 3 and PT 4 groups at 12 and 24 h.ConclusionThe results of this study confirmed the stressful effect of transportation on O. niloticus as well as the transportation stress mitigation effect of NaCl.

Black radish root extract alleviates sodium valproate-induced liver damage via inhibiting mitochondrial membrane potential collapse and oxidative stress in mice

Objective: To explore the effect of black radish (Raphanus sativus L. var niger) root extract on liver enzymes, oxidative stress, and histopathological alterations in mice with sodium valproate-induced hepatotoxicity. Methods: Thirty-two mice were divided into four groups: the control group received drinking water by gavage, the second group was administered with 100 mg/kg of sodium valproate, the third group received 300 mg/kg of black radish root extract, and the fourth group was given both sodium valproate (100 mg/kg) and black radish root extract (300 mg/kg). After 28 days, the mice were euthanized, and serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP), along with liver malondialdehyde (MDA), reactive oxygen species (ROS), mitochondrial parameters, tumor necrosis factor-alpha (TNF-α) gene expression, and histopathological changes were assessed. Results: Sodium valproate caused hepatic damage in mice, characterized by elevated serum levels of liver enzymes, increased MDA and ROS levels and TNF-α gene expression, as well as histopathological alterations. The black radish root extract significantly alleviated sodium valproate-caused hepatic injury by decreasing the serum levels of ALT and AST, MDA, ROS, TNF-α gene expression, as well as mitochondrial impairment, but did not have a significant effect on sodium valproate-induced histopathological changes. Conclusions: The black radish root extract demonstrates protective effects against sodium valproate-induced liver injury, possibly through mitigating oxidative stress, mitochondrial impairment, and inflammatory mediator expression.

Dietary spirulina platensis a promising growth promotor and immune stimulant in broiler chickens

Algae, as a biological component of the environment, holds promise for the development of novel cuisines. This study aimed to appreciate the dietary Spirulina platensis (SP) impact on growth patterns and as an immune stimulant in broilers. SP-fed chicks at 0.5, 1, and 2 g/kg doses significantly improved hematological indices. Also, gas chromatography of fatty acid profile in broiler breast muscles exhibited greater elevation. Serum total proteins, albumin, and globulin levels significantly increased. ElISA (enzyme-linked immunosorbent assay) revealed elevated immunoglobin M, G, and leptin levels as mirrors for immunological response coordination. Reverse transcription polymerase chain reaction (RT-PCR) exhibited depressed tumour necrosis factor-alpha gene expression (TNF-α) in ilial tissue. Gut’s histopathology showed well-developed villi. In conclusion, Spirulina platensis in doses up to 2 g/kg enhances immunity, fatty acid profile, liver function, anti-inflammatory properties, and intestinal absorption of broilers, while doses up to 4 g/kg cause the opposite effect on previous parameters.

DT-13 attenuates inflammation by inhibiting NLRP3-inflammasome related genes in RAW264.7 macrophages

Plant derived saponins or other glycosides are widely used for their anti-inflammatory, antioxidant, and anti-viral properties in therapeutic medicine. In this study, we focus on understanding the function of the less known steroidal saponin from the roots of Liriope muscari L. H. Bailey – saponin C (also known as DT-13) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in comparison to the well-known saponin ginsenoside Rk1 and anti-inflammatory drug dexamethasone. We proved that DT-13 reduces LPS-induced inflammation by inhibiting nitric oxide (NO) production, interleukin-6 (IL-6) release, cycloxygenase-2 (COX-2), tumour necrosis factor-alpha (TNF-α) gene expression, and nuclear factor kappa-B (NFκB) translocation into the nucleus. It also inhibits the inflammasome component NOD-like receptor family pyrin domain containing protein 3 (NLRP3) regulating the inflammasome activation. This was supported by the significant inhibition of caspase-1 and interleukin-1 beta (IL-1β) expression and release. This study demonstrates the anti-inflammatory effect of saponins on LPS-stimulated macrophages. For the first time, an in vitro study shows the attenuating effect of DT-13 on NLRP3-inflammasome activation. In comparison to the existing anti-inflammatory drug, dexamethasone, and triterpenoid saponin Rk1, DT-13 more efficiently inhibits inflammation in the applied cell culture model. Therefore, DT-13 may serve as a lead compound for the development of new more effective anti-inflammatory drugs with minimised side effects.

0118 Association between TNF-α G308A Polymorphism (rs1800629) and Insomnia Vulnerability in Fertile Age Women with Primary Dysmenorrhea

Abstract Introduction Higher tumor necrosis factor-alpha (TNF-α) gene expression has been revealed in the peripheral blood monocytes across the menstrual cycle in patients with primary dysmenorrhea (PDM) than in the unaffected control. Sleep quality is affected by environmental factors, endocrine function abnormalities and genetic factors. The aim of this study was to evaluate the effect of TNF-α G308A polymorphism (rs1800629) on the self-reported sleep, health, menstrual pain severity as well as peri-ovulatory serum TNF-α level in women with and without PDM. Methods Sleep, psychological and physiological characteristics and menstrual pain severity were collected with questionnaires and sleep/menstrual diary within a menstrual cycle from a cohort of 43 PDM women and 42 healthy controls (aged 20-30 years, right-handed, non-smoking and non-shift workers without psychiatric, sleep and neurological comorbidity). Chi-square tests were used to check if the genotype proportions of the investigated gene fitted the Hardy-Weinberg equilibrium (HWE) and to evaluate if the genotypic and allelic distribution is associated with PDM. Chi-square tests or analysis of variance were used to examine the effects of genotype, group and/or genotype x group interaction with the sleep, mind and the body health variables. Results Neither allele frequencies nor genotypes of TNF-α G-308A gene could serve as an independent risk factor of PDM in the Taiwanese population. Circulating serum TNF-α did not correspond with the -308 TNF-α promoter polymorphism, either, which might be due to a multifactorial control process. The TNF-α 308 A allele carriers (28.6%) which was less common than the G allele carriers (71.4%), showed lower Beck depression score, lower bodily pain and better physical health and higher subjective sleep quality. Furthermore, in G allele carriers, the PDM group showed higher insomnia severity and lower sleep duration than that of the control group. Conclusion Present results suggested that TNF-alpha polymorphism conferred no prediction to PDM risk and peri-ovulatory serum TNF-α level. However, our findings were evident that the A allele at the TNFα 308 locus, as compared to the more common G allele, was associated to better health and better sleep, with an indication of a genetic underpinning to their better resistance from the risk of PDM. Support (if any)

Cytokine expression profiles in white blood cells of patients with small fiber neuropathy

BackgroundThe role of cytokines in the pathophysiology, diagnosis, and prognosis of small fiber neuropathy (SFN) is incompletely understood. We studied expression profiles of selected pro- and anti-inflammatory cytokines in RNA from white blood cells (WBC) of patients with a medical history and a clinical phenotype suggestive for SFN and compared data with healthy controls.MethodsWe prospectively recruited 52 patients and 21 age- and sex-matched healthy controls. Study participants were characterized in detail and underwent complete neurological examination. Venous blood was drawn for routine and extended laboratory tests, and for WBC isolation. Systemic RNA expression profiles of the pro-inflammatory cytokines interleukin (IL)-1ß, IL-2, IL-8, tumor necrosis factor-alpha (TNF) and the anti-inflammatory cytokines IL-4, IL-10, transforming growth factor beta-1 (TGF) were analyzed. Protein levels of IL-2, IL-8, and TNF were measured in serum of patients and controls. Receiver operating characteristic (ROC)-curve analysis was used to determine the accuracy of IL-2, IL-8, and TNF in differentiating patients and controls. To compare the potential discriminatory efficacy of single versus combined cytokines, equality of different AUCs was tested.ResultsWBC gene expression of IL-2, IL-8, and TNF was higher in patients compared to healthy controls (IL-2: p = 0.02; IL-8: p = 0.009; TNF: p = 0.03) and discriminated between the groups (area under the curve (AUC) ≥ 0.68 for each cytokine) with highest diagnostic accuracy reached by combining the three cytokines (AUC = 0.81, sensitivity = 70%, specificity = 86%). Subgroup analysis revealed the following differences: IL-8 and TNF gene expression levels were higher in female patients compared to female controls (IL-8: p = 0.01; TNF: p = 0.03). The combination of TNF with IL-2 and TNF with IL-2 and IL-8 discriminated best between the study groups. IL-2 was higher expressed in patients with moderate pain compared to those with severe pain (p = 0.02). Patients with acral pain showed higher IL-10 gene expression compared to patients with generalized pain (p = 0.004). We further found a negative correlation between the relative gene expression of IL-2 and current pain intensity (p = 0.02). Serum protein levels of IL-2, IL-8, and TNF did not differ between patients and controls.ConclusionsWe identified higher systemic gene expression of IL-2, IL-8, and TNF in SFN patients than in controls, which may be of potential relevance for diagnostics and patient stratification.

Antipruritic properties of topical Channa striatus extract on stratum corneum disruption-itch mice model

Itch is an unpleasant sensation that provokes a strong desire to scratch. In some cases, antihistamines are ineffective in treating chronic itch and produce side effects. Channa striatus fish extract, which can alleviate pain, is also used in this experiment to alleviate the itch. This experiment aims to determine the possible antipruritic effect of topical Channa striatus extract (CSE) on stratum corneum disruption (SCD)-itch mice model. Experiments were divided into 6 groups of male ICR mice which were positive, negative, vehicles, 3% CSE, 7% CSE, and 10% CSE groups. Rostral back of mice was induced with chemicals acetone, diethyl ether (1:1) and water (AcetoneEthylWatermethod) daily for five consecutive days. All groups were treated with specific selective treatments. The scratching behaviour of the mice was observed by videotape. The skin moisture and oiliness scoring was measured daily using a skin analyzer. The skin was isolated for RNA extraction and measured for tumour necrosis factor-alpha (TNF- α) gene expression level using real-time RT PCR. All treatment groups of Channa striatus extract cream had shown a significant reduction in scratching time, increase in skin moisture and skin oiliness. The TNF- α gene expression level had inconsistent results and needed a more extensive sample. The Channa striatus extract cream may be able to alleviate itch compared to the control parameters.

Incomplete Protein Predisposes to Cardiometabolic Defect: The Case of Lysine

ObjectivesNon-communicable diseases (NCDs) are common among people with low socioeconomic status who rely on low-quality proteins, such as gluten, that are liming in lysine. This work aims at studying the effect of Lysine Deficiency (LD) on different cardiometabolic parameters to highlight its potential involvement in the development of NCDs. MethodsThe research protocol was approved by the Institutional Animal Care and Use Committee (IACUC) of the American University of Beirut. Sixteen 5 week-old male Sprague-Dawley rats were divided into 2 groups (n = 8). They were maintained on a gluten-protein-based diet: the Control diet (C) contained the recommended level of lysine (9.8 g/kg), and the LD diet had a 60% lower amount. The rats had free access to their corresponding diets over the experimental period of 4 weeks. Hematoxylin and eosin-stained adipocytes from epididymal fats were quantified, and images were used to estimate adipocyte sizes. Total RNA from liver and spleen was isolated and quantitative real-time Polymerase Chain Reaction (PCR) was performed. C-reactive protein (CRP) liver protein expression was estimated using Western Blot. Independent t-tests were conducted to check for differences between groups. Significance was set at a p-value (p) of 0.05. ResultsAt the end of the experiment, the percentage of body fat was similar between groups (p = 0.369), but the epididymal adipose tissue of the LD group showed hyperplasia (p = 0.001) and a decrease in the adipocyte size (p = 0.008). Interestingly, the toll-like receptor 4 (TLR-4) gene was overexpressed in the liver of the LD group (p = 0.016) along with an increase of 85% of tumor necrosis factor-alpha (TNF-α) gene expression (p = 0.027). This was accompanied by an increase of 2.3 fold in CRP protein expression in the liver (p = 0.048). Furthermore, the lipid profile differed between both groups, highlighting a drop of HDL by around 23% and an increase in LDL by 14 folds in the LD group (p < 0.05). ConclusionsLD caused an upsurge in several inflammatory markers and risk factors of cardiovascular diseases. The current data present important evidence about the relationship between lysine deficiency and some metabolic disturbances that increase the risk of NCDs. Funding SourcesThis work was funded by the University Research Board of the American University of Beirut.

Identification of functional tumor necrosis factor-alpha promoter variants associated with Helicobacter pylori infection in the Sudanese population: Computational approach.

BACKGROUND Helicobacter pylori (H. pylori) is a ubiquitous bacterium that affects nearly half of the world’s population with a high morbidity and mortality rate. Polymorphisms within the tumor necrosis factor-alpha (TNF-A) promoter region are considered a possible genetic basis for this disease.AIMTo functionally characterize the genetic variations in the TNF-A 5’-region (-584 to +107) of Sudanese patients infected with H. pylori using in silico tools.METHODSAn observational study was carried out in major public and private hospitals in Khartoum state. A total of 122 gastric biopsies were taken from patients who had been referred for endoscopy. Genomic DNA was extracted. Genotyping of the TNF-A-1030 polymorphism was performed using PCR with confronting two-pair primer to investigate its association with the susceptibility to H. pylori infection in the Sudanese population. Furthermore, Sanger sequencing was applied to detect single nucleotide polymorphisms in the 5’-region (-584 to +107) of TNF-A in H. pylori-infected patients. Bioinformatics analyses were used to predict whether these mutations would alter transcription factor binding sites or composite regulatory elements in this region. A comparative profiling analysis was conducted in 11 species using the ECR browser and multiple-sequence local alignment and visualization search engine to investigate the possible conservation. Also, a multivariate logistic regression model was constructed to estimate odds ratios and their 95% confidence intervals for the association between TNF-A-1030, sociodemographic characteristics and H. pylori infection. Differences were statistically significant if P < 0.05. Statistical analyses were performed using Stata version 11 software.RESULTSA total of seven single nucleotide polymorphisms were observed in the TNF-A 5’-region of Sudanese patients infected with H. pylori. Only one of them (T > A, -76) was located at the in silico-predicted promoter region (-146 to +10), and it was predicted to alter transcription factor binding sites and composite regulatory elements. A novel mutation (A > T, +27) was detected in the 5’ untranslated region, and it could affect the post-transcriptional regulatory pathways. Genotyping of TNF-A-1030 showed a lack of significant association between -1030T and susceptibility to H. pylori and gastric cancer in the studied population (P = 0.1756) and (P = 0.8116), respectively. However, a significant association was detected between T/C genotype and H. pylori infection (39.34% vs 19.67%, odds ratio = 2.69, 95% confidence interval: 1.17-6.17, P = 0.020). Mammalian conservation was observed for the (-146 to +10) region in chimpanzee (99.4%), rhesus monkey (95.6%), cow (91.8%), domesticated dog (89.3%), mouse (84.3%), rat (82.4%) and opossum (78%).CONCLUSIONComputational analysis was a valuable method for understanding TNF-A gene expression patterns and guiding further in vitro and in vivo experimental validation.

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.

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.

Spherical Gold Nanoparticles: Small Interfering RNA Delivery in Regulation of the Tumor Necrosis Factor-Alpha Gene Expression

Proinflammatory cytokines are signaling molecules that are expelled from immune cells like macrophages and other types of cells. Tumor necrosis factor-alpha (TNF-α) is overexpressed during inflammation caused by inflammatory diseases. Therefore, the regulation of TNF-α has a key role in inflammation. The use and target delivery of small interfering RNAs (siRNAs) provide many effectual treatment benefits in the regulation of gene expression in cells. In this study, we used siRNA nanoparticle conjugates in the regulation of gene expression and inflammation. We first prepared safe fusion ribonucleic acid interference carrier, spherical nucleic acid nanoparticle conjugates (SNA-NCs), to enhance the perforation of siRNA into the macrophages and their ability to target TNF-α gene regulation. Furthermore, the suppression of the TNF-α gene was monitored after curing macrophages by SNA-NCs. Gene expression was carried out by real-time polymerase chain reaction in cells and the levels of TNF-α were investigated by the enzyme-linked immunosorbent assay (ELISA) method. This study indicated that the SNA-NCs were safe and very stable. TNF-α siRNA could significantly regulate gene expression in cells to form SNA-NCs. The results indicated that TNF-α gene expression downregulated to 93.40% ± 1.45%, 66.06% ± 0.95%, and 35.76% ± 1.09% in the presence of 0.1, 1, and 10 nM siRNA, respectively. The proliferation of macrophages and subsequently expression of TNF-α were significant for the formation of inflammation. These findings showed that the use of SNA-NC siRNA might ameliorate the inflammatory disease by suppression of gene expression and functional activity of macrophage generation.

Replacing dietary antibiotics with 0.20% l-glutamine and synbiotics following weaning and transport in pigs.

Dietary antibiotic use has been limited in swine production due to concerns regarding antibiotic resistance. However, this may negatively impact the health, productivity, and welfare of pigs. Therefore, the study objective was to determine if combining dietary synbiotics and 0.20% l-glutamine would improve pig growth performance and intestinal health following weaning and transport when compared with traditionally used dietary antibiotics. Because previous research indicates that l-glutamine improves swine growth performance and synbiotics reduce enterogenic bacteria, it was hypothesized that supplementing diets with 0.20% l-glutamine (GLN) and synbiotics (SYN; 3 strains of Lactobacillus [1.2 × 10^9 cfu/g of strain/pig/d] + β-glucan [0.01 g/pig/d] + fructooligosaccharide [0.01 g/pig/d]) would have an additive effect and improve pig performance and intestinal health over that of dietary antibiotics. Mixed-sex pigs (N = 226; 5.86 ± 0.11 kg body weight [BW]) were weaned (19.4 ± 0.2 d of age) and transported for 12 h in central Indiana. Pigs were blocked by BW and allotted to one of two dietary treatments (5 to 6 pigs per pen): antibiotics (positive control [PC]; chlortetracycline [441 ppm] + tiamulin [38.5 ppm]), no antibiotics (negative control [NC]), GLN, SYN, or the NC diet with both the GLN and SYN additives (GLN + SYN) fed for 14 d. From day 14 post-weaning to the end of the grow-finish period, all pigs were provided common antibiotic-free diets. Data were analyzed using PROC GLIMMIX and PROC MIXED in SAS 9.4. Overall, haptoglobin was greater (P = 0.03; 216%) in NC pigs compared with PC pigs. On day 13, GLN and PC pigs tended to have reduced (P = 0.07; 75.2% and 67.3%, respectively) haptoglobin compared with NC pigs. On day 34, the jejunal goblet cell count per villi and per millimeter tended to be greater (P < 0.08; 71.4% and 62.9%, respectively) in SYN pigs compared with all other dietary treatments. Overall, jejunal mucosa tumor necrosis factor-alpha (TNFα) gene expression tended to be greater (P = 0.09; 40.0%) in NC pigs compared with PC pigs on day 34. On day 34, jejunal mucosa TNFα gene expression tended to be greater (P = 0.09; 33.3%, 41.2%, and 60.0%, respectively) in GLN pigs compared with SYN, GLN + SYN, and PC pigs. Although it was determined that some metrics of pig health were improved by the addition of GLN and SYN (i.e., haptoglobin and goblet cell count), overall, there were very few differences detected between dietary treatments and this may be related to the stress load incurred by the pigs.

Effects of dietary olive leaf extract on intestinal immune-related gene expressions in common carp, Cyprinus carpio

This study aimed to investigate the effects of dietary olive leaf extract (OLE) on intestinal immune-related genes expression of tumor necrosis factor alpha (TNFa), interleukin 1 beta (IL1b), lysozyme (LYS), and mucin2 (MUC2). For this purpose, common carp (~15 g) were fed with 0 (control), 0.1 (OLE-0.1), 0.5 (OLE-0.5) and 1 (OLE-1) % OLE diets for eight weeks. The fish were sampled after one and eight weeks to study intestinal TNFa, IL1b, LYS, and MUC2 gene expressions. The results showed that dietary OLE administration significantly up-regulated intestinal TNFa gene expression after one (all OLE-treated groups) and eight (OLE-0.5) weeks. Moreover, OLE-0.1 and OLE-1 groups showed up-regulated intestinal IL1b expression, after one week, all the OLE-treated fish had significantly higher intestinal IL1b expression, after eight weeks. OLE had no significant effects on LYS gene expression after one week, but OLE-0.1 and OLE-0.5 had significantly higher gene expressions after eight weeks. OLE-0.1 and OLE-1 had significantly lower MUC2 gene expression after one week, but all OLE-treated fish had significantly higher MUC2 gene expression after eight weeks. In conclusion, dietary 0.1-0.5% OLE supplementation is suitable to support common carp intestinal health.

MP84-08 TUMOR NECROSIS FACTOR ALPHA IS INCREASED AFTER CAVERNOUS NERVE INJURY AND IMPAIRS REGENERATION OF NITRERGIC NEURONS AND MODULATES PENILE SMOOTH MUSCLE TONE

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology (MP84)1 Apr 2020MP84-08 TUMOR NECROSIS FACTOR ALPHA IS INCREASED AFTER CAVERNOUS NERVE INJURY AND IMPAIRS REGENERATION OF NITRERGIC NEURONS AND MODULATES PENILE SMOOTH MUSCLE TONE Hotaka Matsui*, Nikolai A. Sopko, Jefferey D. Campbell, Xiaopu Liu, Emmanuel Weyne, Maarten Albersen, Fabio Castiglione, Johanna L. Hannan, and Bivalacqua Trinity J Hotaka Matsui*Hotaka Matsui* More articles by this author , Nikolai A. SopkoNikolai A. Sopko More articles by this author , Jefferey D. CampbellJefferey D. Campbell More articles by this author , Xiaopu LiuXiaopu Liu More articles by this author , Emmanuel WeyneEmmanuel Weyne More articles by this author , Maarten AlbersenMaarten Albersen More articles by this author , Fabio CastiglioneFabio Castiglione More articles by this author , Johanna L. HannanJohanna L. Hannan More articles by this author , and Bivalacqua Trinity JBivalacqua Trinity J More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000000976.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Erectile dysfunction is induced after cavernous nerve (CN) injury by inflammation in pelvic ganglia. We examined levels of tumor necrosis factor alpha (TNFA), after bilateral CN injury (BCNI) in vivo, and effect of exogenous TNFA (eTNFA) on neurite outgrowth from major pelvic ganglion (MPG) ex vivo. Additionally, we evaluated effect of TNFA signal inhibition to penile smooth muscle tone with TNFA receptor knockout mice (TNFRKO). METHODS: 70 male rats underwent sham or BCNI. MPGs of sham rats were collected after 48 hours. BCNI groups had MPG harvested at 6 h, 12 h, 24 h, 48 h, 7 days, or 14 days after surgery. We performed qPCR to examine gene expression of TNFA and Western blot to examine protein amount of TNFA in MPGs. Immunofluorescence(IF) for TNFA was performed in MPG. For ex vivo study, MPGs were harvested from control rats and cultured in Matrigel with eTNFA (0, 10, 20, 30 ng/mL). Neurite lengths from MPGs were measured 72 h after culture. MPGs were processed for qPCR of Tyrosine hydroxylase (TH) and neuronal nitric oxidase (nNOS). IF was assessed for TH and nNOS in MPG. To examine effect of TNFA inhibition on MPG and penis after BCNI, wild type mice (WT) and TNFRKO were randomized to undergo sham or BCNI. MPGs were harvested 48 hours after surgery to examine gene expression of nNOS and TH by qPCR. Penises were used to examine smooth muscle tone to electrical field stimulation (EFS). RESULTS: TNFA’s gene/protein expression was increased in MPGs after BCNI, with peaks at 2 and 7 days, respectively. IF showed TNFA localized to perivascular areas at 48 h and neuronal cell body at 14 days after BCNI. eTNFA inhibited neurite outgrowth in a dose-dependent fashion, and impaired gene expression of nNOS in MPG (p<0.05). MPGs cultured with eTNFA showed significantly shorter neurite lengths of nNOS+ neurons than TH+ neurons (p<0.01), while this difference was not seen in controls. TNFRKO showed enhanced gene expression of nNOS compared to WT (p<0.01). Penises of WT showed enhanced smooth-muscle contraction to EFS, compared to TNFKO (p<0.01). TNFRKO had enhanced relaxation response to EFS after maximal contraction, compared to WT (p<0.01). CONCLUSIONS: TNFA was increased following BCNI and inhibited neurite outgrowth from MPGs in a dose-dependent fashion. eTNFA lowered gene and protein expression of nNOS. TNFRKO mice showed enhanced gene expression of nNOS and enhanced penile smooth muscle relaxation. These results suggest that TNFA may selectively inhibit nNOS+ neurons, and impairs penile smooth muscle tone. Source of Funding: NIH © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 203Issue Supplement 4April 2020Page: e1267-e1267 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hotaka Matsui* More articles by this author Nikolai A. Sopko More articles by this author Jefferey D. Campbell More articles by this author Xiaopu Liu More articles by this author Emmanuel Weyne More articles by this author Maarten Albersen More articles by this author Fabio Castiglione More articles by this author Johanna L. Hannan More articles by this author Bivalacqua Trinity J More articles by this author Expand All Advertisement PDF downloadLoading ...

Pharmacological inhibition of G9a/GLP restores cognition and reduces oxidative stress, neuroinflammation and β-Amyloid plaques in an early-onset Alzheimer's disease mouse model.

The implication of epigenetic mechanisms in Alzheimer’s disease (AD) has been demonstrated in several studies. UNC0642, a specific and potent inhibitor of methyltransferase activity G9a/GLP (G9a-like) complex, was evaluated in the 5XFAD mouse model. UNC0642 treatment rescued 5XFAD cognition impairment, reduced DNA-methylation (5-mC), increased hydroxymethylation (5-hmC), and decreased the di-methylation of lysine 9 of histone H3 (H3K9me2) levels in the hippocampus. Increases in the Nuclear Factor erythroid-2-Related Factor 2 (NRF2), Heme oxygenase decycling 1 (Hmox1) gene expression, and diminution in Reactive Oxygen Species (ROS) were also reported. Moreover, neuroinflammatory markers, such as Interleukin 6 (Il-6), Tumor necrosis factor-alpha (Tnf-α) gene expression, and Glial fibrillary acidic protein (GFAP) immunofluorescence were reduced by UNC0642 treatment. An increase in Nerve growth factor (Ngf), Nerve growth factor inducible (Vgf) gene expression, Brain-derived neurotrophic factor (BDNF), and Synaptophysin (SYN) were found after UNC0642 treatment. Importantly, a reduction in β-amyloid plaques was also observed. In conclusion, our work demonstrates that the inhibition of the G9a/GLP complex by UNC0642 delivered significant neuroprotective effects in 5XFAD mice, point out G9a/GLP as a new target for AD.

Effects of lavender (Lavandula angustifolia) extract inclusion in diet on growth performance, innate immunity, immune-related gene expression, and stress response of common carp, Cyprinus carpio

In the present study, the effects of dietary lavender (Lavandula angustifolia) extract (LE) on growth performance and immune, antioxidant, and stress responses in common carp, Cyprinus carpio were investigated. For this, four diets containing 0, 0.5, 1.0 and 1.5% LE were offered to triplicate groups of fish (25.8 ± 1.66 g) for 70 days, and then the fish were exposed to a 3-h crowding stress. Growth performance, blood leukocyte count, plasma proteins and innate immune parameters, and head kidney cytokines’ gene expression were studied after the 70-day feeding trial (before the crowding stress). Plasma cortisol, glucose, superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) were measured before and after the crowding stress. Results showed that dietary LE had no significant effects on the fish growth performance, blood leukocyte differential count, plasma albumin and total protein, and head kidney interferon gamma-1 gene expression. Blood leukocyte and plasma globulin, alternative complement and total immunoglobulin significantly increased in the fish fed with 1.0–1.5% LE diets compared to the fish fed with the control diet (0% LE). The plasma lysozyme activity significantly increased along with dietary LE supplemental levels. The lowest expression of interlukin-1 beta gene was related to the 1.0% LE treatment. Tumor necrosis factor-alpha gene expression significantly decreased along with increase in dietary LE levels and the lowest expression was related to the fish fed with 1.5% LE diet. The LE administration led to significant increase in interlukine-10 gene expression and the highest expression was related to the fish fed with 1.0% and 1.5% LE diets. The fish fed with 1.5% LE diet had significantly higher transforming growth factor-beta gene expression compared to the control group. Liver SOD and CAT activities showed significant increases along with dietary LE supplementation levels. After the crowding stress, hepatic SOD decreased in the control group, but not the LE-supplemented groups. Stress led to significant decrease in hepatic CAT activities in all treatments, but CAT activity increased along with the LE levels with highest activity in the fish fed with 1.5% LE. The stress led to significant increase in hepatic MDA in the treatments control and 0.5% LE, but not 1.0–1.5% LE groups. The stress led to significant increase in plasma cortisol and glucose levels in all treatments but dietary LE levels significantly decreased these parameters. Dietary LE supplementation at levels of 1.0–1.5% is recommended for common carp to suppress stress, inflammation and oxidative conditions and augment immune responses in the fish.

Effect of Energy Drinks on Rat Hippocampus and the Possible Neuroprotective Role of Sesame Oil: Histological, Immunohistochemical and Molecular Study

AbstractBackground: Energy drinks (EDs) consumption has in-creased dramatically over the past years and several adverse effects have been reported. Sesame oil (SO) is a potent antioxidant and anti-inflammatory dietary source for human health.Aim of Study: The present study was assigned to investigate the possible neuroptotecive role of SO on energy drinks-induced hippocampal damage.Material and Methods: Twenty adult male albino rats were divided into four groups; five rats in each (control, SO, EDs and EDs+SO). Routine histological examination was done, together with toluidine blue stain for dark neurons (DN) and glial fibrillary acidic protein (GFAP) immunostaining for astrocytes. Gene expression of catalase and tumor necrosis factor alpha (TNF-a) was analyzed using real-time polymerase chain reaction (PCR) and histomorphometric analysis was done.Results: Histological examination of the hippocampus from EDs group showed shrunken pyramidal and granule cells, vacuolation and large pericellular spaces. There was significant decrease in neuronal thickness, increase in the number of DN and GFAP area percent compared to control group. Significant decrease in catalase gene expression was observed along with increase in TNF-a gene expression. Concomitant SO administration significantly improved the previously mentioned changes.Conclusion: Energy drinks administration induced mor-phological and morphometric changes, deficient antioxidant capacity and inflammatory response in rat hippocampus. Sesame oil administration alleviated these adverse changes possibly due to its antioxidant and anti-inflammatory prop-erties.

Eicosapentaenoic Acid Improves Hepatic Metabolism and Reduces Inflammation Independent of Obesity in High-Fat-Fed Mice and in HepG2 Cells.

The prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing worldwide, concurrent with increased obesity. Thus, there is urgent need for research that can lead to effective NAFLD prevention/treatment strategies. Omega-3 polyunsaturated fatty acids (n-3 PUFAs), including eicosapentaenoic acid (EPA), improve inflammation- and dyslipidemia-related metabolic disorders; however, mechanisms mediating the benefits of n-3 PUFAs in NAFLD treatment are less understood. We previously reported that EPA reversed obesity-induced hepatic steatosis in high-fat (HF)-fed B6 mice. Utilizing a combination of biochemical analyses of liver tissues from HF and HF-EPA-fed mice and a series of in vitro studies in tumor necrosis factor-alpha (TNF-α)-stimulated HepG2 cells, we dissect the mechanistic effects of EPA in reducing hepatic steatosis, including the role of EPA-targeted microRNAs (miRNA). With EPA, hepatic lipid metabolism was improved in HF-EPA mice, as indicated by decreased protein and messenger RNA (mRNA) levels of fatty acid synthase (FASN) and acetyl-CoA carboxylase (Acaca) gene, and increased mRNA levels for the peroxisome proliferator activated receptor-α (Pparα), and carnitine palmitoyltransferase (Cpt) 1a and 2 genes in the HF-EPA mice. Additionally, inflammation was reduced, as shown by decreased tumor necrosis factor-alpha (Tnfα) gene expression. Accordingly, EPA also significantly reduced FASN and ACACA mRNAs in human HepG2 cells. Glycolysis, estimated by extracellular acidification rate, was significantly reduced in HepG2 cells treated with EPA vs. vehicle. Furthermore, we identified several miRNAs that are regulated by EPA in mouse liver, including miR-19b-3p, miR-21a-5p, and others, which target lipid metabolism and inflammatory pathways. In conclusion, our findings provide novel mechanistic evidence for beneficial effects of EPA in NAFLD, through the identification of specific genes and miRNAs, which may be further exploited as future NAFLD therapies.

A Potent Antioxidants Property of Moringa Oleifera on Mycotoxin Induced Oxidative Stress in Rats

This work aimed to evaluate the probable protective effect of Moringa Oleifera on mycotoxins induced liver damage and oxidative stress in rats. Thirty male albino rats were classified randomly into three equal groups. Group1 (normal control): Rats fed with ordinary ration. Group2(Mycotoxin): Rats fed with ordinary diet only for first 7 days and from day 8 rats fed mixed ration contain 50 gram of grinded plant (total aflatoxins 4.2 ppb) + 50 gram of grinded fish (total aflatoxins 42.14 ppb) + 50 gram of ordinary diet (free of aflatoxins) for 4 weeks. Group3 (Mycotoxin+ Moringa): Moringa Oleifera (350 mg/kg b.w) was administered orally allover the experimental period (5 weeks). Also, rats were fed with ordinary ration only for first 7 days and from day 8 rats fed mixed ration that mentioned above in group 2. At the end of the experiment liver tissue specimens were isolated and analyzed for the determination of reduced Glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), L-malondialdehyde (L-MDA) and tumor necrosis factor-alpha (TNF-α) gene expression. The obtained results showed a significant up-regulation of TNF-α gene expression level and marked increase in hepatic SOD, CAT activities and L-MDA of mycotoxin rats group. However, liver GSH concentration was markedly decreased as compared to control group. Moringa protected mycotoxins induced liver damage in rats caused a significant improvement of all previous parameters. Conclusively, MoringaOleiferatreatment exerts a protective effect against liver damage and oxidative stress in liver of mycotoxin intoxicated rats through free radical scavenging and anti-inflammatory activities as well as regenerating endogenous antioxidants defense system mechanisms.