TNF-α Research Articles

Sol-gel synthesis and cytotoxicity evaluation of selenium-doped cerium oxide nanoparticles for biomedical applications.

Over the past few years, ovarian cancer is the second most commonly diagnosed cancer among women. Despite the widespread knowledge of its prevalence, the curative measures and survival rates for ovarian cancer have not improved significantly, making it a challenging condition. Nanotechnology has become increasingly prominent in the field of cancer treatment. Previous studies showed both cerium oxide nanoparticles (CONPs) and selenium (Se) had anti-cancer. Therefore, doping selenium into CONPs may exhibit a more significant anti-cancer effect on ovarian cancer cells. Cerium nitrate hexahydrate, sodium selenite, and gelatin were employed for the production of CONPs and Se-doped CONPs. The EDX, XRD, and TEM/PSA imaging were employed to investigate the structural characteristics and morphology of the synthesized Se-doped CONPs. The reactive oxygen species (ROS) level and TNF, IL-6, and IL-1B gene expression were evaluated after inoculating A2780 human epithelial ovarian carcinoma (HEOC) with Se-doped CONP. Statistical analysis was conducted using ANOVA, followed by Bonferroni's t-test for multiple group comparisons. Se-doped CONPs had IC50 of 113 and 49 PPM after 24 and 48 h, respectively. In addition, Se-doped CONPs with concentrations of 50 and 100 PPM significantly reduced to ROS levels in the HEOC cell line. Also, 50 and 100 PPM Se-doped CONPs lead to significantly reduced TNF, IL-6, and IL-1B gene expression compared to the control group in the HEOC cell line. Our study showed the potential anti-cancer effects of Se-doped CONPs on ovarian cancer cell lines.

Association between TNFα-308 G/A and IFN-γ+874A/T Polymorphisms with Oral Lichen Planus

Background: Oral lichen planus (OLP) is a chronic disease that presents with inflammation and has a global prevalence of 0.1-4%. Lesions of the disease occur in the oral mucosa, gums, and rarely in the palate. This study aimed to investigate the relationship between this disease and TNFα-308G/A and IFN-γ+874A/T polymorphisms. Methods: In this case-control study, oral mucosal samples were collected from 50 healthy subjects, and 50 OLP patients presented to the Kerman Faculty of Dentistry were enrolled using a simple sampling method. Subsequently, we used the amplification refractory mutation system polymerase chain reaction (ARMS-PCR) technique followed by sequencing to determine the presence of TNFα-308G/A and IFN-γ+874A/T polymorphisms in cases and controls. Results: Compared to the control group, the prevalence of A and GA alleles of the TNF gene was higher than the prevalence of G and GG alleles in OLP patients, while the AA genotype of the gene was not found in OLP patients. Regarding IFN gene polymorphism, the relationship between the T allele and the risk of disease was discovered, but it was not statistically significant (P value: 0.068). Conclusion: Although there is a strong relation between the A allele of TNF-α (-308G/A) polymorphism and the risk of OLP, this association between IFN-γ+874A/T genotype and the disease was not strong enough to predict the possibility of developing OLP.

Hubungan Polimorfisme Gen Interleukin 10 - 819 C/T Pada Kejadian Preeklampsia Ibu Hamil di Puskesmas Dalu X B Tanjung Morawa Deli Serdang

Preeclampsia is one of the disorders during pregnancy, characterized by increased blood pressure and proteinuria, causing an increase in morbidity and mortality in the mother and fetus. Interleukin 10 (IL10) is called a T helper type 2 (Th2) cytokine which is important because it acts as an anti-inflammatory which functions for the maintenance and development of pregnancy and forms immunity to inhibit the secretion of T helper 1 (Th1) cytokines such as IL 6, TNF alpha and INF Gamma. Genotypic variation can provide information about individual differences in the secretion of IL-10 and the tendency to the incidence of preeclampsia. The IL 10 - 819 C/T gene polymorphism is located on chromosome 1q32.2 distal to the promoter area and is a transcription factor of the IL 10 gene. The aim of this study was to determine the relationship between 10-819 C/T Interleukin Gene Polymorphisms in the incidence of preeclampsia in pregnant women. The method used to determine the IL 10 819 C/T polymorphism with RFLP PCR. Results. The distribution of IL 10-819 C/T genotypes in the control group had 5 people (23.73%) of TT genetic variation, 15 people with CT (65.21%) and 3 people with CC genetic variation (13.04%). ). In the Preeclampsia group, there were 7 people (30.43%) with genetic variation of TT and there were 10 people (43.47%) with genetic variation of CT as many as 6 people (26.08%) with CC variation. There was no correlation between interleukin gene polymorphism 10-891 C/T with the incidence of preeclampsia in pregnant women at Dalu X B Health Center Tanjung Morawa, Deli Serdang

The effects of time-restricted eating versus habitual diet on inflammatory cytokines and adipokines in the general adult population: a systematic review with meta-analysis

BackgroundTime-restricted eating (TRE) may facilitate weight loss, but its impact on inflammation remains unclear. Chronic inflammation can detrimentally increase risk of obesity-associated comorbidities. ObjectivesThe aim of this systematic review was to synthesize and determine the effects of TRE on cytokine and adipokines (C-reactive protein [CRP], TNF alpha [TNF-α], interleukin-6 [IL-6], leptin, and adiponectin) in adults. MethodsPubMed, Scopus, Cochrane CENTRAL, and Web of Science were systematically searched for randomized controlled trials (RCTs) and non-RCTs to determine the effects of TRE on cytokines and adipokines in adults up to 23 June, 2023. Risk of bias was assessed using risk of Bias 2 tool for RCTs and the ROBINS-I for non-RCTs. The standardized mean differences (SMDs) and their 95% confidence intervals (CIs) were estimated with the DerSimonian–Laird method through random-effect models. The PRISMA recommendations were followed. ResultsA total of 25 studies (13 RCTs, 12 non-RCTs) involving 936 participants were included. The pooled SMD for the effect of TRE compared with the control group on cytokines and adipokines was −0.11 (95% CI: −0.33, 0.12; I2 = 19.7%; n = 10 comparisons) for CRP; −0.25 (95% CI: −0.47, −0.03; I2 = 0%; n = 11 comparisons) for TNF-α; −0.09 (95% CI: −0.39, 0.21; I2 = 16.4%; n = 8 comparisons) for IL-6; −0.81 (95% CI: −1.37, −0.24; I2 = 65.3%; n = 5 comparisons) for leptin; and 0.07 (95% CI: −0.40, 0.54; I2 = 56.9%; n = 6 comparisons) for adiponectin. ConclusionsTime-restricted eating may be an effective approach to reduce TNF-α and leptin levels in the general adult population.This review was registered at PROSPERO as CRD42022358162.

Molecular Characterisation of Blood Microbiome in Patients with Ankylosing Spondylitis and Healthy Controls.

In human and animal studies, ankylosing spondylitis (AS) has been increasingly linked to changes in the microbial inhabitants in the human body (microbiome). These studies have primarily now concentrated on the microbial communities that live in the gastrointestinal tract. However, evidence suggests that various molecular techniques can be used to detect microbial DNA in blood circulation. This DNA might be an unknown reservoir of biomarkers with the potential to track alterations in the microbiomes of remote locations, such as the gut. To this end, we compared the presence and identity of microbial DNA in blood samples taken from ankylosing spondylitis patients to healthy control subjects by amplifying and sequencing the bacterial 16S rRNA variable region four. The study's design is a case study based on the presence and identity of bacterial DNA in the blood of Ankylosing spondylitis (AS) patients (n = 10) and healthy control subjects (n = 10) was investigated by amplifying and sequencing the bacterial 16S rRNA gene. Blood concentrations of the cytokines TNF alpha, IL-17A, and IL-23 were determined by the Human Magnetic Luminex Screening, and data were analysed using an Unpaired T-test. Using PCR amplification, 8 of 10 AS patients (80%) and 8 of 10 healthy control samples (80%) had microbial 16S rRNA in their blood. At the phylum level, Proteobacteria (Control = 48.5%, AS = 52%), Firmicutes (Control = 27.8%, AS = 26.1%), Actinobacteria (Control = 15.4%, AS = 10.7%), and Bacteroidetes (Control = 6.5%, AS = 10%) dominated the blood microbiome. A two-tailed Mann-Whitney test found that Ankylosing Spondylitis was associated with significantly elevated Bacteroides (P < 0.05), Prevotella (P < 0.001), and Micrococcus (P < 0.01), and significantly reduced levels of Corynebacterium 1 (P < 0.001), Gemella (P < 0.01), and Alloprevotella (P < 0.05), compared to healthy controls. Additionally, it was shown that the presence of the Prevotella genus was highly positively correlated with higher levels of TNF-alpha (P < 0.05; r = 0.8) in AS patients' blood. This article reveals that a blood microbiome exists in healthy individuals and identifies particular taxa modulated in disease. These blood-derived signatures indicate that this field needs more research and may be helpful as disease biomarkers.

Ganoderic acid C2 exerts the pharmacological effects against cyclophosphamide-induced immunosuppression: a study involving molecular docking and experimental validation

Triterpenoids, as the main active ingredient of Ganoderma lucidum fermented extract, exert multiple pharmacological activities, including immunomodulatory properties. Our study aimed to reveal the pharmacological effects and potential mechanisms of Ganoderic acid C2 (GAC) against cyclophosphamide (CY)-associated immunosuppression. Target genes were collected from several public databases, including the DisGeNET, Comparative Toxicogenomics Database, GeneCards, and PharmMapper. STRING database was used to construct the protein–protein interaction of network. Subsequently, molecular docking was carried out to visualize the protein-GAC interactions. Experimental validations, including ELISA and qRT-PCR were performed to confirm the pharmacological activities of GAC on CY-induced immunosuppression model. A total of 56 GAC-related targets were identified to be closely associated with CY-induced immunosuppression. Enrichment analyses results revealed that these targets were mainly involved in immune and inflammatory response-related pathways. STAT3 and TNF were identified as the core targets of GAC. Molecular docking indicated that GAC combined well with STAT3 and TNF protein. In addition, animal experiments indicated that GAC improved immunity as well as STAT3 and TNF genes expression in CY-induced immunosuppression, which further verified the prediction through bioinformatics analysis and molecular docking. We successfully revealed the potential therapeutics mechanisms underlying the effect of GAC against CY-induced immunosuppression based on the combination of bioinformatics analysis, molecular docking, and animal experiments. Our findings lay a theoretical foundation for the in-depth development and utilization of Ganoderma lucidum fermentation product in the future, and also provide theoretical guidance for the development of innovative drugs that assist in improving immunity.

Effect of P-glycoprotein inhibitor (Carvedilol) on pharmacological and hematological effects of its substrate (methotrexate) on pregnant and lactating rat mothers

Sixty albino rats (40 female rats and 20 males) were allocated into four dosing groups administered orally carvedilol (T1) 0.72mg/kg, methotrexate (T2) 0.36mg/kg, combined doses (T3) carvedilol + methotrexate and control group (Distilled water) for 2 months in male and 2 weeks in female rats before mating and after copulation and approval of pregnancy, dosing continued in female groups during pregnancy and lactation periods. Half of the animal groups were euthanized one day before parturation to determine P-gp concentration in the placenta and liver of pregnant mothers. In contrast, the other half was left for parturition and lactation to study the effects of carvedilol (P-gp inhibitor) on methotrexate (P-gp substrate) when given alone and in combination with some pharmacological, hematological, and biochemical parameters in lactating mothers and pups at the end of lactation. The pharmacological results showed the highest significant antinociceptive responses both in the early and late phase in the T3 (CV+MTX) followed by the T2 (MTX) group. In contrast, the T1 (Cv) animal group showed nearly resembling results as that of the control group in the early and late phases of the formalin test. The written test also recorded the same result as the Formlin test, indicating a significantly higher analgesic effect in T3 followed by T2 in comparison to T1 and control groups. The TNF alpha results supported the anti-inflammatory effect, recording a significant decline mainly in the T3 and T2 groups. The hematological results recorded significantly more reduction in WBC levels in mother and their pups in comparison with T1 and control groups. The serum electrolyte levels (Na+, K+, Ca+2) recorded in all treated groups and pups showed a significant increase in potassium levels in the T1 and T2 mother rat groups. The P-glycoprotein (P-gp) concentration measured in liver and placenta of euthanized pregnant mother before delivery recorded potent inhibition of P-gp in T1 &amp; T3 groups which might altered pharmacokinetic and pharmacodynamic effects of substrate by carvedilol that considered a potent P-gp inhibitor drugs, this might explain our results of increase in pharmacological effect of methotrexate and their side effect on electrolyte concentration and hematology in combined group of CV+ MTX. Keywords: Carvedilol, Methotrexate, p-gp inhibitor, Anti-inflammatory, Hematology, Electrolyte.

Investigation of corticosteroid-NSAID prodrugs for TNF-α and IL6 receptors, their characterization, single crystal XRD and DFT studies: A combined experimental and theoretical approach

In continuation of our research towards the synthesis of steroidal derivatives, we have synthesized corticosteroids-NSAIDs derivatives 3i-3p as potent anti-inflammatory agents. The products obtained have been characterized using elemental analysis, spectroscopic techniques (FT-IR, 1H NMR, 13C NMR, UV–Vis and mass spectrometry) and product 3k has also been characterized with the help of single crystal X-ray crystallography. The inhibition of pro-inflammatory cytokines is a key approach to abate severe inflammation. In this work a series of corticosteroids-NSAIDs derivatives are reported whose activity were performed on RAW264.7 cell line and have been identified as potential TNF alpha and IL6 inhibitors. Our work demonstrates that these corticosteroid derivatives may be useful for the development of anti-inflammatory agents. Among these derivatives, 11,17-dihydroxy-3,20-diketo-pregn-1,4-diene-21-yl-2-(1,8‑diethyl-1,3,4,9-tetrahydropyrano[3,4-β]indol-1-yl)acetate (3j) was identified as the most potent anti-inflammatory agent, which showed significant anti-inflammatory activity by inhibiting the LPS-induced pro-inflammatory mediator TNF-α and IL6 in a dose-dependent manner without any cytotoxicity.

SAT425 Effects Of Endocrine-disrupting Polychlorinated Biphenyls On Adolescent Microglial Cytokine Expression And Responses To Secondary Inflammatory Challenge

Abstract Disclosure: S.L. Kissinger: None. A.J. Fitzgibbon: None. R.A. Mejia: None. M.R. Bell: None. Polychlorinated biphenyls (PCBs) are a group of ubiquitous environmental endocrine-disrupting chemicals (EDCs), and exposure is associated with endocrine, neuronal, and immune dysfunction in various tissue types. Innate immune cells of the nervous system called microglia are critical in responding to infection and injury, and disruption of these functions is associated with neurodegenerative disease and mental health disorders. In addition, microglia are known to regulate developmental processes during neonatal and adolescent periods, in hormone-sensitive and age- and sex-differentiated ways. Work in our lab has shown that early life PCBs can affect microglia function in ways that depend on the age and sex of the individual rat in vivo. Ongoing work uses in vitro approaches to demonstrate direct effects on microglia. In primary culture, preliminary data suggest that acute PCB exposure causes neonatal microglia to become hyperresponsive to a secondary challenge; however, effects in adolescent microglia are unknown. To test the hypothesis that PCB exposure directly alters microglia activity in an age- and sex-specific manner, we determined the effects of PCBs on cytokine gene expression in acutely isolated microglia, with or without a secondary lipopolysaccharide (LPS) challenge. Adolescent Sprague-Dawley rats underwent transcardial saline perfusion, brains were removed and homogenized into a single cell suspension with collagenase and mechanical disruption, and debris was removed. Microglia, identified as CD11b+ cells, were collected by magnetic bead isolation and plated in enriched media for 24 hours prior to experimentation. Cells were exposed to one of three concentrations of PCB 153 (0.1 µM, 1 µM, 10 µM), DMSO vehicle, or DMEM/F12 media control for 18 hours, followed by a 4-hour LPS challenge (0.5 µg/mL) or PBS vehicle control. RNA was isolated and converted to cDNA for qPCR analysis of Tnf and Il1b gene expression, analyzed by two-way (exposure x challenge) ANOVAs within both male and female cells. Results indicate that 1.0 µM PCB 153 exposure caused an increase in expression of both cytokines, more so in females than males. Preliminary results (n = 3) indicate that the 10 µM dose may also blunt the later LPS response. These results confirm that microglia are directly sensitive to PCBs, however they illustrate that cells may show differential responses depending on the stage of development. Continued work will explore acute cell toxicity and confirm release of cytokines. Adolescent microglia dysfunction from PCB exposure may be a mechanism of adolescent-onset disorders, or increased risk of neurodegenerative conditions triggered by additional challenges later in life. Presentation: Saturday, June 17, 2023

S3239 Paradoxical Pustular Psoriasis Secondary to TNF Alpha Inhibitor Use in Ulcerative Colitis

S3239 Paradoxical Pustular Psoriasis Secondary to TNF Alpha Inhibitor Use in Ulcerative Colitis

Plasma Levels of Organochlorine Pesticides and Tumor Necrosis Factor-Alpha: A Potential Risk Factor for Developing Acquired Aplastic Anemia in the North Indian Population.

Background Pesticide exposure might have a contributory role in the development ofacquired aplastic anemia (AA). However, the precise mechanisms of pesticide-induced AA remain unknown. In this case-control study, we conducted a comparative analysis of plasma levels of organochlorine pesticides (OCP) and tumor necrosis factor-alpha (TNF-alpha) between Indian patients diagnosed with AA and an age- and sex-matched control group. Methods This is an observational case-controlstudy conducted at a tertiary care hospital in North India. In this study, 90 subjects were included, out of which 45 were diagnosed with AA according to the criteria of the International Agranulocytosis and Aplastic Anemia Study. Cases were compared with 45 controls. A trained interviewer gave all study subjects a questionnaire to collect data regarding demographic details, exposure to pesticides, and clinical history.Physical examinationand routine laboratory investigations of each subject were performed. Both cases and controls were tested for their plasma levels of organochlorines as per established protocol by gas chromatography-mass spectrometry. TNF-alpha level was measured by enzyme-linked immunosorbent assay in each subject. Results There was a significant increase in plasma levels of delta hexachlorocyclohexane(delta HCH) (p = 0.02) and heptachlor (p = 0.00) in patients with AA as compared to controls. We observed nonsignificant trends towards higher levels of beta HCH (p = 0.643), aldrin (p = 0.399), and p,p'-Dichlorodiphenyltrichloroethane (p,p'-DDT) (p = 0.453) in patients with AA when compared to the controls. There were significantly higherTNF-alpha levels (p = 0.024) in cases as compared to the controls. Conclusion Our study concludes that patients with AA exhibited higher levels ofdelta-HCH, heptachlor, andTNF-alpha in comparison to the control group. There is a significant positive correlation of TNF alpha with OCPs (alpha HCH, lindane, delta HCH, heptachlor, aldrin, p,p'- DDD, and methoxychlor pesticides). These organochlorines may have accumulated in the fatty tissue of bone marrow because of their lipophilic nature. This suggests that they might have served as a neoantigen to trigger an increase in TNF-alpha production, which may have led to disrupted bone marrow function through cell-mediated immunity, leading to AA.

DNA Hypomethylation in the TNF-Alpha Gene Predicts Rheumatoid Arthritis Classification in Patients with Early Inflammatory Symptoms.

DNA-methylation data were screened to select candidate CpGs to design qMSP assays. Eight assays were developed and tested on two early inflammatory arthritis cohorts. Logistic regression and bootstrapping were used to demonstrate the added value of the qMSP assays. Differentially methylated CpG data were screened for candidate CpG, thereby meeting the qMSP assay requirements. The top CpG candidate was in the TNF gene, for which we successfully developed a qMSP assay. Significantly lower DNA-methylation levels were observed in RA (p < 4 × 10-9), with a high predictive value (OR < 0.54/AUC < 0.198) in both cohorts (n = 127/n = 157). Regression using both datasets showed improved accuracy = 87.7% and AUC = 0.944 over the model using only clinical variables (accuracy = 85.2%, AUC = 0.917). Similar data were obtained in ACPA-negative patients (n = 167, accuracy = 82.6%, AUC = 0.930) compared to the clinical variable model (accuracy = 79.5%, AUC = 0.892). Bootstrapping using 2000 datasets confirmed that the AUCs for the clinical+TNF-qMSP model had significant added value in both analyses. The qMSP technology is robust and can successfully be developed with a high specificity of the TNF qMSP assay for RA in patients with early inflammatory arthritis. It should assist classification in ACPA-negative patients, providing a means of reducing time to diagnosis and treatment.

Сytokine expression and production in severe cases of SARS-CoV-2 infection

Most respiratory viral infections proceed in mild form including COVID-19. Gowever, some patients experience severe systemic inflammation, tissue damage, acute respiratory distress syndrome, and cytokine storm with potentially lethal outcomes. The cytokines have been thought to play an important role in immunopathology of viral infection. However, an excessive immune response, manifesting as massive release of pro-inflammatory cytokines, may cause immune damage in the body. Production and release of the cytokines in healthy individuals presumes a significant balance of inflammatory and homeostatic factors. Meanwhile, in the case of COVID-19 disease, uncontrolled increased production of cytokines often occurs with fatal consequences for patients. The aim of this work was to study the level of IL-1β, IL-18 and TNFα gene expression, as well as production of these cytokines at the level of mucous membranes of the upper respiratory tract, in particular in oral cavity, in patients with severe COVID-19 disease.&#x0D; The present study included patients who recovered from severe COVID-19. The control group consisted of conditionally healthy individuals. Expression levels of the IL-1β, IL-18, and TNFα genes were determined by RT-PCR. The levels of IL-1β, IL-18 and TNFα protein production were determined by multiplex enzyme immunoassay.&#x0D; The expression levels of IL-1β, IL-18 were reduced at the onset of the disease, as well as in the midpoint of the COVID-19 disease, but increased on the 30th day. The protein production of these cytokines was also reduced in the first days from the onset of the disease. The levels of pro-inflammatory TNFα cytokine was high at the onset of the disease. The level of TNFα production at the onset of the disease was also higher relative to the control group. Subsequently, the TNFα gene expression levels decreased upon progression of the disease.&#x0D; Thus, the increased expression level of pro-inflammatory cytokines may be explained by the fact that the S protein of the SARS-CoV-2 virus induces increased expression of these cytokines in human monocytes. Meanwhile, appropriate protein levels remain low, especially on day 1 from the onset of the disease. Thus, one may conclude that the virus triggers pyropotosis, however, within 15-30 days from the onset of the disease, when viral replication is already minimal.

Immunobiology of lymphotoxin: role in a mouse model of multiple sclerosis

Complex immunobiology of lymphotoxin (LTα) is due to multiple modalities of signal transduction, involving a soluble homotrimer and membrane-bound heterotrimers that engage at least three different receptors. While LTα is crucial for the formation and maintenance of secondary lymphoid organs, its overproduction is observed in autoimmune diseases such as rheumatoid arthritis and multiple sclerosis. Initially, LTα was considered pathogenic in the development of experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis, as demonstrated by the resistance of mice with genetic LTα inactivation to EAE induction. However, conflicting observations arose when EAE was induced in RAG1-deficient mice that underwent adoptive bone marrow transfer from LTα-deficient mice, thereby calling into question previous conclusions about the role of LTα in EAE development.&#x0D; This study aimed to investigate the role of LTα in MOG35-55-induced EAE using mice deficient in LTα or its membrane receptor, LTβR. LTα knockout mice used here were designed to avoid the artifact involving TNF gene downregulation in myeloid cells, which occurred in the conventional LTα knockout mice.&#x0D; Surprisingly, LTα-deficient mice with normal TNF expression developed EAE clinically comparable to wild-type mice. Conversely, genetic inactivation of LTβR delayed EAE onset. However, during the later stages of the disease, LTβR deletion exacerbated clinical symptoms of EAE.&#x0D; These findings demonstrate that the involvement of LTα in EAE development is more complex than previously estimated, and that LTβR exhibits diverse functions depending on the disease stage: pathogenic at the early stage and protective at the later stages of EAE.

Mechanism of herb pair containing Astragali Radix and Spatholobi Caulis in the treatment of myelosuppression based on network pharmacology and experimental investigation

Ethnopharmacological relevanceThe Astragali Radix and Spatholobi Caulis herb pair (ARSC) is one of the most commonly used herbal combinations for bone marrow suppression. According to traditional Chinese medicine, Astragali Radix strengthens the spleen and replenishes qi, while Spatholobi Caulis is a hematinic agent that promotes blood circulation and enrichment. The compatibility of the two helps the body to tonify the spleen and kidneys and compensate for visceral deficiencies. However, the multi-target mechanism of ARSC in bone marrow suppression has remained largely unknown. Aim of the studyThe aim of this study is to explore the key targets and signaling pathways of the traditional Chinese herbal pair ARSC for the treatment of bone marrow suppression. Materials and methodsThe active components of ARSC and targets for myelosuppression were screened using network databases. Cytoscape 3.8.0 was used to construct compound-target, compound-disease-target and protein-protein interaction (PPI) networks. Go-function and pathway enrichment analyses were performed to explore the potential mechanism. In vivo animal experiments were conducted to verify the molecular mechanisms. ResultsThe 36 active compounds were identified from the ARSC, and a total of 108 genes involved in myelosuppression were screened. VEGFA, IL6, TNF, JUN, STAT3, PTGS2, CASP3 and MMP9 genes were identified as potential drug targets in the PPI network analyzed by CytoHubba. Enrichment analysis indicated that ARSC may treat myelosuppression through various biological processes, such as apoptosis, TNF-α signaling pathway via NF-κB, PI3K/AKT/mTOR signaling pathway, IL6/JAK/STAT3 signaling pathway, P53 signaling pathway and G2/M checkpoint signaling pathway. The results of the experiment showed that the aqueous extract of ARSC significantly alleviated myelosuppression, reduced the apoptosis rate of bone marrow cells, upregulated the mRNA expression levels of TNF-α, IL-6 and VEGF, and promoted NF-κB phosphorylation in myelosuppressed mice. ConclusionsThis study identified the active components and relevant mechanisms of ARSC in the treatment of myelosuppression. Our findings predicted that ARSC could treat bone marrow suppression through multiple components, multiple targets and multiple pathways. Pharmacological experiments showed that ARSC alleviated fluorouracil-induced myelosuppression by reducing the apoptosis rate of bone marrow cells and regulating the TNF-α/NF-κB signaling pathway.

Upregulation of miR-21 and pro-inflammatory cytokine genes IL-6 and TNF-[formula omitted] in promoting a pro-tumorigenic microenvironment in canine mammary carcinomas

This study evaluated the gene expression of the pro-inflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in canine mammary tumors (CMTs), and correlated them with gene expression of miRNAs expected to regulate the secretion of pro-inflammatory cytokines within the tumor microenvironment (TME). Furthermore, gene expression of cytokines and miRNAs involved in tumor cell proliferation and invasion (i.e. miR-21; miR-124; miR-145) were correlated with tumor proliferation index (Ki67 index) to determine the prognostic value in CMTs. Twenty-six canine mammary samples were used, including 22 CMTs and 4 control samples. MiR-21, IL-6 and TNF-α were upregulated in mammary carcinomas compared with controls (p < 0.05). MiR-146b was downregulated in CMTs compared with control cases (p < 0.05). IL-6 expression showed a significant positive correlation with miR-21 and a negative correlation with miR-146b; while, TNF-α gene expression was positively correlated with miR-21 and miR-145 in mammary carcinomas. In carcinomas, the Ki67 index correlated positively with gene expression of IL-6 and miR-21 and negatively correlated with miR-145 and miR-146b. Specifically, gene expression of IL-6 and miR-21 was positively correlated with ki67 index >33.3%, whereas, expression of miR-145 and miR-146b was negatively correlated with ki67 index <33.3%. Results reinforce the concept of interaction between tumor cells and inflammatory cells within the TME, with a central role of IL-6 and TNF-α. Since the upregulation of miR-21 reflects the gene overexpression of interleukins and the high proliferation index of tumor cells, this miRNA may be considered a biomarker with prognostic value in CMTs.

Tumor Necrosis Factor-α (-308G>A) Gene Polymorphism and Its Association with Asthma and Atopy Status.

Asthma is one of the most prevalent chronic lung diseases that afflict genetically predisposed individuals. Certain cytokine gene polymorphisms have been associated with asthma. Tumor necrosis factor-alpha (TNF-α) is a potent inflammatory cytokine that can modulate nonspecific inflammation to influence asthma. This study aimed to define the relationship between the TNF gene polymorphism at position -308 and asthma susceptibility, as well as atopic and non-atopic asthma. Using polymerase chain reaction with sequence-specific primers, we investigated genotype frequencies and alleles of a polymorphic gene coding for TNF-α in 86 pediatric patients with asthma and 470 healthy controls of the same race. Seventy-four patients underwent a skin prick test. The homozygous AA variant (-308, rs1800629) was the most common genotype among patients, accounting for 63.3% of all cases. In contrast, homozygous GG (-308) was significantly less prevalent in the patient group compared to the control group. TNF A (-308) allele frequency was 85.5% among asthma patients and 16.6% among healthy controls. The genotype and allele frequencies of TNF (-308 A>G, rs1800629) did not differ between atopic and non-atopic asthma. In conclusion, TNF (-308) AA and AG genotypes are associated with asthma susceptibility in Iranian children, although there was no significant difference in polymorphism between atopic and non-atopic asthma and no difference in asthma severity groups.

Repetitive mild traumatic brain injury-induced neurodegeneration and inflammation is attenuated by acetyl-L-carnitine in a preclinical model.

Repetitive mild traumatic brain injuries (rmTBI) may contribute to the development of neurodegenerative diseases through secondary injury pathways. Acetyl-L-carnitine (ALC) shows neuroprotection through anti-inflammatory effects and via regulation of neuronal synaptic plasticity by counteracting post-trauma excitotoxicity. This study aimed to investigate mechanisms implicated in the etiology of neurodegeneration in rmTBI mice treated with ALC. Adult male C57BL/6J mice were allocated to sham, rmTBI or ALC + rmTBI groups. 15 rmTBIs were administered across 23 days using a modified weight drop model. Neurological testing and spatial learning and memory assessments via the Morris Water Maze (MWM) were undertaken at 48h and 3months. RT-PCR analysis of the cortex and hippocampus was undertaken for MAPT, GFAP, AIF1, GRIA, CCL11, TDP43, and TNF genes. Gene expression in the cortex showed elevated mRNA levels of MAPT, TNF, and GFAP in the rmTBI group that were reduced by ALC treatment. In the hippocampus, mRNA expression was elevated for GRIA1 in the rmTBI group but not the ALC + rmTBI treatment group. ALC treatment showed protective effects against the deficits displayed in neurological testing and MWM assessment observed in the rmTBI group. While brain structures display differential vulnerability to insult as evidenced by location specific postimpact disruption of key genes, this study shows correlative mRNA neurodegeneration and functional impairment that was ameliorated by ALC treatment in several key genes. ALC may mitigate damage inflicted in the various secondary neurodegenerative cascades and contribute to functional protection following rmTBI.

Lactococcus garvieae ile Enfekte Archocentrus centrarchus’ta Bağışıklıkla İlişkili Enzim Gen Etkileşimleri

Unsuitable environmental conditions and low host resistance in the aquatic environment are the most important factors in fish diseases. Bacterial diseases in fish can be treated with different preparations. The aim of this study is to determine the disease symptoms, expression levels of TNF and IL genes and some biochemical parameter changes in Archocentrus centrarchus by using herbal extract for the treatment of the disease caused by Lactococcus garvieae bacteria. Gene expression was performed by Real-Time PCR and enzyme activities were determined by spectrophotometric analysis method. It was observed that the expression level of TNF gene was 2.5 times more effective in the antibiotic (oxytetracycline) group and 14 times more effectively in the extract application. It was found that the antibiotic group had a close expression level compared to the control at the IL gene expression level, while the extract group expressed 5.5 times more. Glutathione s-transferase activity in the extract group increased significantly compared to the control and antibiotic groups. Superoxide dismutase activity was significantly higher in the extract group compared to the antibiotic and bacterial groups. It can be said that oxidative stress caused by bacterial infection is tried to be eliminated more in the extract group. It was determined that the andiz root extract which at the patent stage, which was used for the first time in the treatment of A. centrarchus fish infected with L. garvieae, one of the most important bacterial agents in the field of aquaculture, is applicable.

P01.04.B LONG-TERM COGNITIVE TOXICITY IN SMALL CELL LUNG CANCER POPULATION: EARLY CLINICAL, NEUROIMAGING, AND SEROLOGICAL BIOMARKERS

Abstract BACKGROUND Small cell lung cancer patients (SCLC) are treated with platinum-based chemotherapy, thoracic radiation, and some of them will also receive prophylactic cranial irradiation (PCI). Following therapy, nearly 45% of SCLC will exhibit moderate to severe cognitive impairment (CI). The aim of our study was to identify early biomarkers including clinical, serological and changes in brain structure associated with an increased risk of CI in SCLC population. MATERIAL AND METHODS Fifty-two SCLC were prospectively recruited from January 2019-December 2022 at our institution and evaluated at diagnosis using a neuropsychological battery (NPS), a structural MRI (T1-weighted), and serological markers including: a) cytokines (interleukin 1, 2, 6, 8, 10 and TNF alpha) and b) single nucleotide polymorphisms (SNP) of apolipoprotein E (APOE), multidrug resistance protein (MDR1) and brain-derived neurotrophic factor (BDNF). They were also clinically and NPS evaluated at 1-month following chemotherapy-before PCI, at 3-6 months, at one-year and at 2-3 years post-PCI. Voxel-based morphometry (T1-VBM) was used to analyze gray matter (GM) integrity. RESULTS Of the 52 patients included, 70% were male, median age was 65 years (46-82). Most (94%) had an ECOG PS ≤1. Half had hypertension (52%) and dyslipidemia (46%); one third (36%) moderate alcoholism and 25% were diabetic. Nearly half (54%) had limited stage and received cisplatin-based chemotherapy (52%). One third (29%, n=15) exhibited brain metastasis (BM): 4 at baseline (8%) and the other during follow-up (21%). Regarding smoking, median packs per year was 45 (12-100). Twenty-nine (56%) received cranial radiation, most of them (n=22, 42%) PCI (25Gy). Median overall survival was 18 months (2.5, 71); twenty-one (40%) were alive at the end of the study.Nearly half of our cohort (48%, n=25) exhibited CI: 31% (n=16) at baseline and another 17% (n= 9) during follow-up. CI group performed significantly worse in all the domains explored, especially in verbal memory (p &amp;lt; 0.001) and executive functioning (p &amp;lt; 0.001). Only smoking habit resulted significantly different between groups: CI patients were heavier smokers (p=0.04). VBM analysis showed that CI group exhibited significant GM damage in anterior and mid cingulate, superior, and inferior temporal gyrus, precuneus and fusiform gyrus. GM volume positively correlated with measures of cognitive functioning at baseline (all correlations, p &amp;lt;0.04). We found no differences regarding cytokines levels or SNPs of APOE, MDR1 and BDNF between groups. CONCLUSION This study confirms that nearly half of the SCLC population exhibit CI. Of greater interest, however, is the fact that most of them already exhibit CI before therapy that seem to correlate with GM alterations previously described in this population. In addition, smoking seems to be the most important clinical risk factor.