Alpha Gene Expression Research Articles

Deciphering Folate Receptor alphaGene Expression and mRNA Signatures in Ovarian Cancer: Implications for Precision Therapies

Deciphering Folate Receptor alphaGene Expression and mRNA Signatures in Ovarian Cancer: Implications for Precision Therapies

The Impact of Acute Ammonia Nitrogen Stress on Serum Biochemical Indicators and Spleen Gene Expression in Juvenile Yellowfin Tuna (Thunnus albacares).

The presence of ammonia nitrogen in water has a significant impact on the serum and spleen of fish, potentially leading to changes in substances such as proteins in the serum while also causing damage to the immune function of the spleen. To investigate the effects of ammonia nitrogen (NH3-N) stress on juvenile yellowfin tuna (Thunnus albacares), this study established three NH3-N concentrations, 0, 5, and 10 mg/L, denoted as L0, L1, and L2, respectively. Serum and spleen samples were collected at 6, 24, and 36 h. The effects of different NH3-N concentrations and exposure times on the physiological status of juvenile fish were investigated by analyzing serum biochemical indices and splenic gene expression. The results indicate that in the L1 group, the serum high-density lipoprotein cholesterol (HDL-C), triglycerides (TG), complement 3 (C3) and complement 4 (C4) levels, and acid phosphatase (ACP) activity showed a trend of initially increasing and then decreasing. In the L2 group, the serum low-density lipoprotein cholesterol (LDL-C), HDL-C, and C4 levels and ACP activity also displayed an initially rising and then declining trend, while TG, C3, and creatinine (CRE) levels and alkaline phosphatase (AKP) activity showed an upward trend. In the L1 group, glutathione peroxidase 1b (GPX1b), interleukin 10 (IL-10), interleukin 6 receptor (IL-6r), and tumor necrosis factor alpha (TNF-α) gene expression levels in the spleen exhibited a trend of first increasing and then decreasing. In the L2 group, IL-10, IL-6r, tumor necrosis factor beta (TNF-β), caspase 2 (casp2), and caspase 9 (casp9) gene expression levels in the spleen also showed an initial increase followed by a decrease. When NH3-N levels are below 5 mg/L, it is recommended to limit stress exposure to no more than 36 h for the juvenile fish. For concentrations ranging from 5 to 10 mg/L, stress should be strictly controlled to within 24 h. Exposure to high NH3-N levels may affect biochemical indicators such as serum lipid metabolism, immunity, and metabolism in juvenile fish, and may damage the expression of antioxidant, immune gene, and apoptosis factors in the spleen. This study aims to deepen our understanding of the effects of NH3-N on juvenile tuna, with the goal of establishing effective water quality monitoring and management strategies. This will ensure the quality of aquaculture water, reduce the harm caused by NH3-N to juvenile yellowfin tuna, and enhance aquaculture efficiency and product quality.

The Influence of Circulating Exosomes Derived From Younger and Older Donors on Hypoxia-Inducible Factor 1 Alpha Gene Expression and P21 Protein in Cord Blood Hematopoietic Stem Cells

The Influence of Circulating Exosomes Derived From Younger and Older Donors on Hypoxia-Inducible Factor 1 Alpha Gene Expression and P21 Protein in Cord Blood Hematopoietic Stem Cells

Influence of Ascorbic Acid on Di-(2-Ethylhexyl) Phthalate-induced Ovarian Gene Alterations in Pubertal Female Wistar Rats

Background Di-(2-ethylhexyl) phthalate (DEHP), a plasticizer compound affecting female reproduction, leads to scenarios, such as polycystic ovarian syndrome (PCOS) and infertility through oxidative stress (OS) mechanisms. Ascorbic acid (AA) is one of the antioxidants in infertility issues. Objectives The present study investigates the ameliorative effect of AA on DEHP-induced ovarian toxicity in pubertal female Wistar rats. Materials and Methods Thirty female Wistar rats of four weeks of age were stratified into five groups. Group I was treated with corn oil (Vehicle), groups II and III with low and high dose DEHP, and groups IV and V with low and high dose DEHP+AA were administered for 30 days. Results Increased body weight gain was noted in DEHP groups. Estradiol hormone was considerably reduced, whereas progesterone levels were increased in both low- and high-dose DEHP-treated groups. DEHP+AA groups have shown significant ( p < 0.005) protection of these hormone levels as equal to the control group. The high-dose DEHP group shows an increased, ovarian estrogen receptor (ER) alpha, ER-beta, and progesterone receptor gene expression, and DEHP+AA groups have significantly ( p < 0.005) showed expression similar to the control. OS was noted with decreased superoxide dismutase and increased malondialdehyde expression in Group III (GR III) compared to control, whereas the DEHP+AA treated group significantly protected OS by restoring the expression levels. DEHP-treated groups show elevated levels of both Bcl-2 and BAX which is specific to apoptotic expression and restored by AA treatment ( p < 0.005). Conclusion Evidence suggests that AA may protect against DEHP-induced ovarian toxicity by decreasing OS levels, improving folliculogenesis, and restoring the hormonal with receptor level alterations.

Cyperus peptide SFRWQ inhibits oxidation and inflammation in RAW264.7 cell model

Oxidative stress can induce many diseases. Antioxidant peptides from food sources have the advantages of good safety, high activity, and good absorbability. In this study, a pentapeptide (SFRWQ; SER-PHE-ARG-TRP-GLN) was identified in a protein hydrolysate of Cyperus (Cyperus esculentus L.). Enzyme-linked immunosorbent assay (ELISA), real-time quantitative (qPCR), immunofluorescence and other techniques were used to evaluate the anti-inflammatory and antioxidant effects of SFRWQ. SFRWQ was found to have 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging ability, help increase superoxide dismutase (SOD) and catalase (CAT) levels in RAW264.7 cells, reduce reactive oxygen species (ROS) levels, and decrease tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) gene expression and secretion. The binding score of SFRWQ to recombinant Kelch-like ECH-associated protein 1 (Keap1) was greater than that of TX6. These findings suggest that SFRWQ activates the Keap1-Nrf2 cellular antioxidant signaling pathway. According to metabolomics studies, SFRWQ increased glutathione (GSH), glutathione disulfide (GSSG), and γ-glutamylcysteine levels and decreased the levels of Prostaglandin D2 (PGD2), Prostaglandin E2 (PGE2), and Prostaglandin H2 (PGH2), which are involved in arachidonic acid metabolism, to protect cells from LPS-induced damage. By elucidating the mechanism of action of SFRWQ, we provide a reference for the development of dietary antioxidant peptides.

Abstract 2809: Bifidobacterium species BB_003 identified through metatranscriptomic analysis with experimentally validated antitumor actions

Abstract Introduction & Objectives: A host's microbiome influences both gene expression and phenotype. Whole transcriptome sequencing (WTS) provides information about the microbiome in addition to gene expression. This research probes the correlation between microbial abundance, host gene transcription, and therapeutic efficacy, pinpointing a beneficial microbial strain and its mechanism of action. Materials & Methods: Bladder tissue biopsies from NMIBC patients underwent WTS and subsequent bioinformatic analysis. After in silico evaluation, in vitro assays including WST, quantitative PCR, and immune cell profiling were performed on mouse splenocytes. These assays were followed by in vivo assessment in both subcutaneous and orthotopic murine tumor models. Results: Among 59 biopsies, non-recurrence correlated with heightened presence of Lactococcus species LC_001 and Bifidobacterium species BB_003. Notably, BB_003 was associated with an extended recurrence-free survival (RFS). Gene set enrichment analysis (GSEA) implicated BB_003 in mediating a pro-inflammatory cascade, accentuating macrophages and interferons. In vitro WST assays showed the antitumor potential of BB_003_β, characterized by upregulated pro-inflammatory gene transcription. This strain increased effector CD8 T cells and M2 macrophages. In vivo, direct live BB_003_β introduction into subcutaneous tumors curtailed tumor proliferation. An increase in effector CD8 T cell populations within tumors and elevated cDC1 and M1 macrophage counts in tumor-draining lymph nodes were observed. Intravesical administration of live bacteria markedly decreased bladder weight, amplifying interferon alpha and beta gene expressions compared to controls. Conclusions: According to this study, BB_003_β inhibits tumor growth and is involved in immune response. Our study confirmed the antitumor properties of BB_003_β and suggested that cytokine and immune cell stimulation play a key role in its mechanism of action. Augmented insights warrant further exploration, particularly in synergy with intravesical BCG treatment. Citation Format: Xuan-Mei Piao, Kyungchan Min, Young Joon Byun, Chuang-Ming Zheng, Seon-Kyu Kim, Seong-Hwan Park, Sungmin Moon, Kyeong Kim, Ho Won Kang, Won Tae Kim, Seok Joong Yun, Hansoo Park. Bifidobacterium species BB_003 identified through metatranscriptomic analysis with experimentally validated antitumor actions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2809.

Fructose regulates the pentose phosphate pathway and induces an inflammatory and resolution phenotype in Kupffer cells

Over-consumption of fructose in adults and children has been linked to increased risk of non-alcoholic fatty liver disease (NAFLD). Recent studies have highlighted the effect of fructose on liver inflammation, fibrosis, and immune cell activation. However, little work summarizes the direct impact of fructose on macrophage infiltration, phenotype, and function within the liver. We demonstrate that chronic fructose diet decreased Kupffer cell populations while increasing transitioning monocytes. In addition, fructose increased fibrotic gene expression of collagen 1 alpha 1 (Col1a1) and tissue metallopeptidase inhibitor 1 (Timp1) as well as inflammatory gene expression of tumor necrosis factor alpha (Tnfa) and expression of transmembrane glycoprotein NMB (Gpnmb) in liver tissue compared to glucose and control diets. Single cell RNA sequencing (scRNAseq) revealed fructose elevated expression of matrix metallopeptidase 12 (Mmp12), interleukin 1 receptor antagonist (Il1rn), and radical S-adenosyl methionine domain (Rsad2) in liver and hepatic macrophages. In vitro studies using IMKC and J774.1 cells demonstrated decreased viability when exposed to fructose. Additionally, fructose increased Gpnmb, Tnfa, Mmp12, Il1rn, and Rsad2 in unpolarized IMKC. By mass spectrometry, C13 fructose tracing detected fructose metabolites in glycolysis and the pentose phosphate pathway (PPP). Inhibition of the PPP further increased fructose induced Il6, Gpnmb, Mmp12, Il1rn, and Rsad2 in nonpolarized IMKC. Taken together, fructose decreases cell viability while upregulating resolution and anti-inflammatory associated genes in Kupffer cells.

Protocatechuic acid alleviates TMAO-aggravated atherosclerosis via mitigating inflammation, regulating lipid metabolism, and reshaping gut microbiota.

Trimethylamine-N-oxide (TMAO) is a risk factor for atherosclerosis. As a natural phenolic acid, protocatechuic acid (PCA) is abundant in various plant foods. The present study investigated the effect of PCA on TMAO-aggravated atherosclerosis in ApoE-/- mice. The mice were randomly divided into five groups and fed one of the following five diets for 12 weeks: namely a low-fat diet (LFD), a western diet (WD), a WD + 0.2% TMAO diet (WDT), a WDT + 0.5% PCA diet (WDT + LPCA), and a WDT + 1.0% PCA diet (WDT + HPCA). Results demonstrated that dietary TMAO exacerbated the development of atherosclerosis by eliciting inflammation and disturbing lipid metabolism. The diet with PCA at 1% reduced TMAO-induced aortic plaque by 30% and decreased the levels of plasma pro-inflammatory cytokines. PCA also improved lipid metabolism by up-regulating the hepatic gene expression of peroxisome proliferator-activated receptor alpha (PPARα). In addition, PCA supplementation enhanced fecal excretion of fatty acids and decreased hepatic fat accumulation. PCA supplementation favorably modulated gut microbiota by increasing the α-diversity with an increase in the abundance of beneficial genera (Rikenella, Turicibacter, Clostridium_sensu_stricto and Bifidobacterium) and a decrease in the abundance of the harmful Helicobacter genus. In summary, PCA could alleviate the TMAO-exacerbated atherosclerosis and inflammation, improve the lipid metabolism, and modulate gut microbiota.

PSIX-7 Attenuation of Immune Allergic Response Pathways in Canines with Dermatological Problems Fed a Polyphenol-Rich Food

Abstract Pathogenesis of canine dermatological problems is mediated by multiple effector cells including lymphocytes, eosinophils, and mast cells. We have previously reported a significant increase in the gene expression of interleukin-5 receptor alpha (IL-5RA) in dogs with dermatological problemscompared with controls, as well as a modest increase in C-C chemokine receptor 3 (CCR3), IL-4 and IL-5. We now investigated whether 2 tests foods (one rich in polyphenols) would provide beneficial effects in canines with dermatological problems (n = 8) compared with controls (n = 8). Dogs were pre-fed control food for 28 days and then divided into two groups based on their age, gender and phenotype (Healthy, Derm) followed by a cross-over design. At each phase, dogs were fed for 42 days with test food 1 (TF1) and the second group was fed with a greater polyphenol content Test food 2 (TF2) for 42 days followed by a crossover. Blood samples were collected at the end of pre-feed, end of phase 1, and end of phase 2. The study was approved by IACUC and AWC, Hill’s PNC. RNA was extracted from blood collected in PAXgene tubes and gene expression was investigated using the NanoString nCounter platform. To assess the food effect, a mixed model analysis was performed using treatment (food) as a fixed effect and animal as random effect using JMPPro. TF2 fed dogs decreased pruritus scores, increased the response rate (improvement %) to 30.77% compared with dogs fed TF1. There was a significant decrease in the interleukin-5 receptor alpha (IL-5RA) in dogs fed TF2 when compared with baseline (P < 0.05). IL-5RA is a receptor expressed on eosinophils as well as committed eosinophil precursors in the bone marrow. There was also a decrease in FCRL2 in dogs fed TF2 (P < 0.05). This gene is preferentially expressed in memory B-cells and plasma cells and also involved in B-cell receptor signaling. The generation of plasma cells requires naïve B cell activation to differentiate into antibody secreting cells or memory B cells. Given the lower levels of FCRL2 with TF2, it is possible that the production of subsequent IgE is attenuated. Another signaling molecule that was significantly decreased with TF2 was the innate immune response molecule C3AR1, a receptor for Complement C3a that induces chemotaxis, granule release, as well as superoxide production. TF1 and TF2 significantly decreased CCR3, a receptor on eosinophils and basophils. CCR3 antagonists are effective in reducing allergic inflammation. Our results indicate the important role of eosinophilic allergic response in dermatological disorders in dogs. TF2 appears to have a beneficial effect in attenuating allergy-inducing signaling pathways of innate and adaptive immune responses. Targeting these molecules may be important as part of newer therapeutic approaches, including nutritional strategies, to manage dermatological problems in dogs.

Using combined single-cell gene expression, TCR sequencing and cell surface protein barcoding to characterize and track CD4+ T cell clones from murine tissues.

Single-cell gene expression analysis using sequencing (scRNA-seq) has gained increased attention in the past decades for studying cellular transcriptional programs and their heterogeneity in an unbiased manner, and novel protocols allow the simultaneous measurement of gene expression, T-cell receptor clonality and cell surface protein expression. In this article, we describe the methods to isolate scRNA/TCR-seq-compatible CD4+ T cells from murine tissues, such as skin, spleen, and lymph nodes. We describe the processing of cells and quality control parameters during library preparation, protocols for multiplexing of samples, and strategies for sequencing. Moreover, we describe a step-by-step bioinformatic analysis pipeline from sequencing data generated using these protocols. This includes quality control, preprocessing of sequencing data and demultiplexing of individual samples. We perform quantification of gene expression and extraction of T-cell receptor alpha and beta chain sequences, followed by quality control and doublet detection, and methods for harmonization and integration of datasets. Next, we describe the identification of highly variable genes and dimensionality reduction, clustering and pseudotemporal ordering of data, and we demonstrate how to visualize the results with interactive and reproducible dashboards. We will combine different analytic R-based frameworks such as Bioconductor and Seurat, illustrating how these can be interoperable to optimally analyze scRNA/TCR-seq data of CD4+ T cells from murine tissues.

Effect of cinnamon supplementation on the gut environment and ESR gene expression in the ovaries of laying quail

This study addressed the hypothesis that the inclusion of powdered cinnamon in a laying quail diet could increase the expression of genes related to the antioxidant defense systems in the intestine and improve the intestinal environment, thus promoting balance in the body. We used 144 laying quail (Coturnix japonica) distributed in a completely randomized design with two treatments: no supplementation of cinnamon (NSC) and supplementation of 9g/kg of cinnamon powder (CPS). At the end of the experimental period ovarian and duodenal tissues were collected for analysis of gene expression. Fragments of the central portions of the duodenum and jejunum were also collected for intestinal histological analysis. We observed a significant effect of the inclusion of cinnamon powder in the diet quail on the expression of the estrogen receptor alpha gene (P = 0.0004). The animals from the CPS treatment presented a higher height of villi and greater depth of crypts in the duodenum, and a higher height of villi and villus:cryptratio in the jejunum. The results show that cinnamon supplementation improves the integrity of intestinal tissue structures and development of the structures of the epithelium. Thus improves the productive performance of the birds.

Gene expression of protein kinase AMP-activated catalytic subunit alpha 1 (PRKAA1), solute carrier family 2 member 1 (SLC2A1) and mechanistic target of rapamycin (MTOR) in metformin-treated type 2 diabetes patients with COVID-19: impact on inflammation markers.

The COVID-19 pandemic has resulted in a global health crisis that has severely impacted patients with type 2 diabetes (T2D). T2D patients have a higher risk of experiencing severe COVID-19 symptoms, hospitalization, and mortality compared to patients without diabetes. The dysregulated immune response in T2D patients can exacerbate the severity of COVID-19 symptoms. Insulin therapy, a common treatment for T2D patients, has been linked to increased mortality in COVID-19 patients with T2D. However, metformin, an anti-diabetic medication, has been shown to have anti-inflammatory properties that may mitigate the cytokine storm observed in severe COVID-19 cases. In this study, we investigated how the PRKAA1, SLC2A1, and MTOR genes contribute to inflammation markers in COVID-19 patients with T2D, who were receiving either insulin or metformin therapy. Our findings revealed that metformin treatment was associated with reduced expression of genes involved in Th1/Th17 cell differentiation. These results suggest that metformin could be a potential treatment option for T2D patients with COVID-19 due to its anti-inflammatory properties, which may improve patient outcomes.

Effect of Eight Weeks of Resistance Training on Inflammatory Markers and Antioxidant Indices of Kidney Tissue Following Testosterone Enanthate Abuse in Male Rats

Introduction: Abuse of anabolic steroids can lead to kidney tissue damage.The aim of this study was to investigate the effect of eight weeks of resistance training on inflammatory markers and antioxidant indices of kidney tissue of male rats following testosterone enanthate abuse.
 Methods: In this experimental study, 24 male Wistar rats were randomly divided into three groups: 1) Control, 2) Training and 3) Training + Testosterone. Rats in groups 2 and 3 performed resistance training five sessions a week for eight weeks. Moreover, group 3 received 20 mg/kg of testosterone enanthate intramuscularly three days a week. The activity levels of Superoxide dismutase (SOD) and Glutathione peroxidase (GPx) were measured by spectrophotometric method and the gene expression of Interleukin-6 (IL-6) and Tumor necrosis factor alpha (TNF-α) were measured by Real Time PCR method. The data were analyzed using SPSS version 16 software. To analyze the findings, To analyze the findings, one-way analysis of variance was used (P≥0.05).
 Results: Resistance training caused a significant decrease in GPx activity (P=0.001) compared to the control group. IL-6 gene expression in the training + testosterone group was significantly more than the training group (P=0.04) and the control group (P=0.001). Training + testosterone caused a significant increase in TNF-α gene expression compared to the training group (P=0.02) and the control group (P=0.008). Furthermore, the level of GPx activity in the training + testosterone group was significantly lower than the control group (P=0.001), but no significant difference was observed in the training + testosterone groups and the training group (P=0.93).
 Conclusion: It seems that testosterone abuse along with intense resistance training is associated with a decrease in the function of the antioxidant system and increase in inflammatory markers in the kidney tissue.

Effects of bisphenol A on murine salivary glands and human tumor cell lines

Bisphenol A (BPA) is an endocrine-disrupting chemical with a potential role in endocrine cancers. However, the effects of BPA on the salivary glands have been barely explored. We investigated the impact of in vivo sub-chronic exposure to BPA and its in vitro effects on human salivary gland mucoepidermoid carcinoma cell lines. Male and female mice were exposed to BPA (30 mg/kg/day). Sublingual and submandibular salivary glands from an estrogen-deficiency model were also analyzed. BPA concentration in salivary glands was evaluated by gas chromatography coupled to ion trap mass spectrometry. Immunohistochemical analysis using anti-p63 and anti-α-SMA antibodies was performed on mouse salivary gland tissues. Gene expression of estrogen receptors alpha and beta, P63 and α-SMA was quantified in mouse salivary gland and/or mucoepidermoid (UM-HMC-1 and UM-HMC-3A) cell lines. Cell viability, p63 and Ki-67 immunostaining were evaluated in vitro. BPA disrupted the tissue architecture of the submandibular and sublingual glands, particularly in female mice, and increased the expression of estrogen receptors and p63, effects that were accompanied by significant BPA accumulation in these tissues. Conversely, ovariectomy slightly impacted BPA-induced morphological changes. In vitro, BPA did not affect the proliferation of neoplastic cells, but augmented the expression of p63 and estrogen receptors. The present data highlight a potential harmful effect of BPA on salivary gland tissues, particularly in female mice, and salivary gland tumor cells. Our findings suggest that estrogen-dependent pathways may orchestrate the effects of BPA in salivary glands.

Elevated cardiac hemoglobin expression is associated with a pro-oxidative and inflammatory environment in primary mitral regurgitation

BackgroundPrimary mitral regurgitation (PMR) is associated with oxidative and inflammatory myocardial damage. We reported greater exosome hemoglobin (Hb) in pericardial fluid (PCF) versus plasma, suggesting a cardiac source of Hb. ObjectiveTest the hypothesis that Hb is produced in the PMR heart and is associated with increased inflammation. Methods and resultsHb gene expression for subunits alpha (HBA) and beta (HBB) was assessed in right atria (RA), left atria (LA) and left ventricular (LV) tissue from donor hearts (n = 10) and PMR patient biopsies at surgery (n = 11). PMR patients (n = 22) had PCF and blood collected for macrophage markers, pro-inflammatory cytokines, and matrix metalloproteinases (MMPs). In-situ hybridization for HBA mRNA and immunohistochemistry for Hb-alpha (Hbα) and Hb-beta (Hbβ) protein was performed on PMR tissue. ResultsHBA and HBB genes are significantly increased (>4-fold) in RA, LA, and LV in PMR vs. normal hearts. In PMR tissue, HBA mRNA is expressed in both LV cardiomyocytes and interstitial cells by in-situ hybridization; however, Hbα and Hbβ protein is only expressed in interstitial cells by immunohistochemistry. PCF oxyHb is significantly increased over plasma along with low ratios (<1.0) of haptoglobin:oxyHb and hemopexin:heme supporting a highly oxidative environment. Macrophage chemotactic protein-1, tumor necrosis factor-α, interleukin-6, and MMPs are significantly higher in PCF vs. plasma. ConclusionThere is increased Hb production in the PMR heart coupled with the inflammatory state of the heart, suggests a myocardial vulnerability of further Hb delivery and/or production during cardiac surgery that could adversely affect LV functional recovery.

Oxydative stress in type 2 diabetic patients: involvement of HIF-1 alpha AND mTOR genes expression

Biochemical and genetic mechanisms of oxidative stress (OS) developing in blood of patients with type 2 Diabetes mellitus (T2DM) were studied. Twenty patients with T2DM and 10 healthy persons participated in this study. Lipid peroxidation, the content of protein carbonyls and H2O2 production were measured in blood plasma and erythrocytes as OS biomarkers. Activity of SOD, catalase, and GPx as well as reduced glutathionе (GSH) level in plasma and erythrocytes were estimated. The gene expression of key regulators of oxygen and metabolic homeostasis (HIF-1α and mTOR) in leukocytes was studied. It was found a significant rise of TBARS and protein carbonyls content in plasma as well as of H2O2 production in erythrocytes from patients with T2DM compared to control. The diabetic patients also demonstrated an increase in the SOD and catalase activity in plasma and significantly lower GSH content and GPx activity in erythrocytes compared to the healthy participants. The established marked inhibition of mTOR gene expression and the tendency to an increase in HIF-1α gene expression in leukocytes of patients with T2DM may serve as a protective mechanism which counteracts OS developing and oxidative cell damage. Keywords: HIF-1α, mTOR, oxidative stress, type 2 diabetes mellitus

Bone Marrow-Derived Mesenchymal Stem Cells and Pioglitazone or Exendin-4 Synergistically Improve Insulin Resistance via Multiple Modulatory Mechanisms in High-Fat Diet/Streptozotocin-Induced Diabetes in Rats.

Diabetes mellitus (DM) is a metabolic disease, characterized by hyperglycemia resulting from defects in insulin secretion and/or insulin action. The current study was designed to assess the therapeutic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) alone and in combination with pioglitazone (Pz) or exendin-4 (Ex) in high-fat diet/streptozotocin (HFD/STZ)-induced diabetes in rats. The rats were subjected to the HFD for three weeks before being injected with a single low dosage of STZ (35 mg/kg bw). The animals were assigned to different treatment groups after type II diabetes mellitus (T2DM) induction was confirmed. Severe insulin resistance was verified in untreated HFD/STZ T2DM rats, along with the exaggeration of oxidative stress, inflammation, apoptosis, and autophagy suppression in the adipose tissues. Monotherapy of HFD/T2DM rats with BM-MSCs and Pz or Ex alleviated diabetic complications by increasing insulin sensitivity, decreasing apoptosis and inflammation as evidenced by a decrease in serum tumor necrosis factor-alpha, caspase-3, and nuclear factor-kappa B (NF-κB) genes expression and Janus kinase (JNK) protein expression, and enhancing autophagy as revealed by upregulation in beclin and LC3, as well as peroxisome proliferator-activated receptor-γ coactivator-1 alpha (PGC-1α) genes expression in the adipose tissues. An augmented ameliorative efficacy was recorded in combined treatments. The biochemical and molecular results were confirmed by histological investigation of pancreatic tissues. Combining Pz or Ex with BM-MSCs is a synergistic therapeutic option that reduces insulin resistance and subsequent complications in T2DM via multiple molecular mechanisms.

Investigating the Safety and Efficacy of the Synthetic Drug Herbix on Immune Responses Involved in the Treatment of a Mouse Model of Herpes Simplex Virus.

Herpes simplex virus-1 (HSV-1) infections can cause significant harm to individuals, including blindness, congenital defects, genital herpes, and even cancer, with no definitive cure .so, finding new treatment strategies is crucial. In this study, 25 male BALB/c mice were used to conduct a mouse model of herpes by subcutaneously injecting an HSV-1 suspension (100 µL of 1× PFU/mL). The mice were divided into 5 groups with groups 1 to 3 designated as intervention groups, and groups 4 and 5 serving as positive and negative control groups, respectively. After 2 days of virus inoculation, the mice were treated with different concentrations of Herbix (100, 200, and 300 mg/mL) via subcutaneous injection. Mice Blood samples (0.5 to 1 mL) were taken from the mice before and after the experiments, and after three-week follow-up period, the mice were sacrificed and the spleens were removed for lymphocyte analysis. we found that administration of Herbix at a dose of 300 mg/mL showed the greatest efficacy, characterized by a delay in skin lesion formation, an increment in survival rate and lymphocyte proliferation, upregulation of the gene expression of interferon alpha (IFN-α) and tumor necrosis factor alpha (TNF-α), and an increase in the polarization of cytotoxic and helper T lymphocytes compared to the control group. These results suggest that Herbix at a dose of 300 mg/mL is effective in treating murine herpes and stimulating immune responses, making it a potential candidate for further investigation as an antiherpetic drug.

Fingolimod prevents cognitive impairments following hypoxia-induced neonatal seizure by ameliorating the inflammation and oxidative stress in male and female juvenile rats

Hypoxia-induced neonatal seizure (HINS) is associated with cognitive disorders in later life. Since the neuroinflammation following HINS leads to these deleterious effects, in the current study, the short-term outcomes of Fingolimod (FTY720) treatment as an anti-inflammatory agent was investigated in the rat model of HINS. Seizures were induced in postnatal day (P10) by exposure to 5 % O2 for 15 min. Forty-five minutes after the onset of hypoxia, pups received FTY720 (0.3 mg/kg) or normal saline for 12 consecutive days. Then, behavioral functions of the rats were assessed by commonly used tests. Subsequently, hippocampal tissue sampling at P22-P23 was done for biochemical assessments and real-time PCR. The results showed that FTY720 prevents cognitive impairments and related reduction of malondialehyde (MDA) concentration in the hippocampus of both male and female hypoxic animals. Furthermore, FTY720 administration following HINS could not prevent the decreased brain-derived neurotrophic factor (BDNF) concentration, increased nitric oxide (NO) level and decreased Interleukin-4 (IL-4) gene expression in the hippocampus of both male and female hypoxic animals. Interestingly, FTY720 showed significant interaction with sex in hippocampal Tumor necrosis factor alpha (TNF-α) gene expression and BDNF concentration. Taken together, these results suggest that FTY720 administration in the lactation period can prevent the deleterious effects of HINS on cognition in later life by ameliorating inflammation and oxidative stress in the hippocampus of both male and female juvenile rats.

Lymphotoxin alpha gene expression in Iraqi breast cancer women

Introduction and Aim: Women's breast cancer is the number one cancer documented in Iraq, with cumulative evidence suggesting cytokines, including tumor necrosis factors probably contributing to the development of the disease. Thus, the aim of this study was to assess the contribution of Lymphotoxin alpha gene to breast cancer. Materials and Methods: In this study, 100 women were enrolled; 50 breast cancer patients (25 women diagnosed with breast cancer but without treatment and 25 women who underwent surgery and different treatments); 25 women with benign tumours, and 25 apparently healthy women. Each participant's blood sample (2 ml) was collected in a triazole-containing tube and stored at – 20°C until use. For the detection of Lymphotoxin alpha gene expression, RNA was extracted from blood and subjected to qPCR. Results: The study showed that 52% of breast cancer patients were over 50 years old among which 73% had attained menopause. The expression of the LTA gene was increased in breast cancer patients before treatment and in those with benign tumours (BT), which suggests that LTA might enhance the development of tumour cell breast cancer. Conclusion: LTA expression may be associated to the development of breast cancer in women.