Concentrations Of Inflammatory Cytokines Research Articles

Association of peripheral and cerebral inflammatory markers with suicidal deaths in Indian population.

Globally, a suicide occurs every 43 seconds. In India, approximately 465 suicides took place daily in 2022, with a suicide rate of 12.4 per 100,000 population, exceeding the global rate of 9. This study aims to investigate differences in cytokine levels, including vascular endothelial growth factor (VEGF), between individuals who died by suicide and those who did not. Additionally, it aims to explore any pattern between plasma and cerebrospinal fluid (CSF) levels of these analytes. Concentrations of inflammatory cytokines, i.e., Interleukins: IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-13, Tumor Necrosis Factor-α and VEGF in plasma and CSF were analyzed among suicide death subjects (N = 6, cases) and non-suicidal death subjects (N = 59, controls). All analytes were quantified using a customized multiplex (9-plex) kit, with 50 µl of plasma and CSF in duplicates. Significantly higher levels of IL-6 (p < 0.001) and IL-10 (p < 0.05) were observed in the plasma of the suicidal group. However, no statistically significant changes were found in the rest of the analytes between the two groups. In CSF, only VEGF levels were significantly lower in the suicidal group (p < 0.01). In cases, significant positive correlations were observed between plasma and CSF levels of IL-1β, IL-4, IL-6, and IL-10, whereas in controls, only IL-8 demonstrated a significant positive correlation, indicating a moderate relationship between these cytokines systemic and central concentrations. Dysregulation of inflammatory biomarkers appears to play a crucial role in the pathophysiology of suicide. We conclude that higher levels of IL-6 are present in the plasma of suicide completers compared to non-suicidal deceased individuals.

Effect of metformin on the level of aqueous humor inflammatory cytokines in patients with cataract

This study investigated the content of inflammatory cytokines in the aqueous humor (AH) of cataract patients with type 2 diabetes (T2DM) and explored the effect of metformin on the level of cytokines. AH was collected from patients undergoing phacoemulsification and intraocular lens implantation in Peking University Third Hospital. Levels of cytokines were measured by Cytometric Bead Assay (CBA) Flex Set. Differences in level of AH cytokines were compared between patients using metformin and non-metformin medicine as blood sugar control drug for T2DM and age-related cataract patients without T2DM. A total of 67 patients were included, including 19 healthy controls, 33 patients in the metformin group, and 15 patients in the non-metformin group. The results showed that IL-6 levels were significantly higher in the non-metformin group than the metformin group and the healthy control group (p = 0.019 and 0.014, respectively). IFN-γ levels were also significantly higher in the non-metformin group than the metformin group and the healthy control group (p = 0.031 and 0.003, respectively). The levels of IL-10 in non-metformin group were significantly higher than those in the healthy control group (p = 0.008), whereas the levels of IL-10 showed no significant difference between metformin group and healthy controls. Metformin can reduce the level of cytokines in AH to a certain extent in cataract patients combined with T2DM. It is suggested that metformin may have preventive and therapeutic effects on the development of age-related cataract.

Anti-cystitis glandularis action exerted by glycyrrhetinic acid: bioinformatics analysis and molecular validation.

Cystitis glandularis (CG) is a chronic hyperplastic disorder of the bladder, and the available clinical drug therapy is insufficient currently. Glycyrrhetinic acid (GA), a bioactive compound extracted from the roots of Glycyrrhiza glabra, is found with beneficial actions, including anti-inflammatory and anti-oxidative effects. We previously reported that GA relieves CG symptoms in animal model, implying the potential application of GA to treat CG. However, the action mechanisms of GA against CG remain unclear. In this study, we aimed to identify the pivotal targets and therapeutic effects of GA through integrated bioinformatics analysis and experimental validation. Integrated bioinformatics analysis screened eleven potential therapeutic targets for GA against CG, and seven pivotal targets were identified subsequently. Enrichment gene analysis revealed GA exhibiting biological activities against CG via regulating multiple pharmacological targets and molecular pathways associated with inflammatory reaction and oxidative stress. Molecular docking computation revealed potent affinity and interaction between GA and prostaglandin-endoperoxide synthase2(PTGS2) and mucin 1 (MUC1) proteins in CG. To validate biochemically, increased mRNA and protein expressions of PTGS2 and MUC1 were observed in human CG samples. Compared to CG mice, GA-treated CG mice exhibited reduced inflammatory cytokine contents and downregulated PTGS2 and MUC1 mRNA and protein levels. These integrated findings suggest the potential therapeutic effects of GA against CG via the regulation of targeting genes and pathways. However, further studies are necessary to perform and facilitate the clinical application of GA for treating CG.

The inflammatory profiling in a cohort of older patients suffering from cognitive decline and dementia.

During aging, there is a progressive impairment of immune cell function that triggers the production of pro-inflammatory cytokines causing the so-called "inflammaging". Frailty represents a condition of increased vulnerability to stresses and reduced homeostatic reserve reflecting not only health status but also biological age. In older subjects without dementia, we showed that markers of inflammaging were differently associated with chronological age than with frailty. This study analysed the same markers in older people with cognitive decline and/or dementia. The cohort consisted of 776 community-dwelling older people: 235 patients with mild cognitive impairment, 63 with Alzheimer's disease (AD), 175 with mixed dementia (MD), and 303 subjects without cognitive decline. Plasma concentrations of inflammatory cytokines and peripheral markers of neuroinflammation were analysed by next-generation ELISA. After adjustment for age, sex, frailty, education and Apolipoprotein E genotype, only interleukin-10, interleukin-1β, and neurofilament light chain were associated with the risk for AD and MD. Moreover, interleukin-6 showed a weak association only with AD. Our data showed similar associations between AD and MD, supporting the concept that late-onset dementia is a complex outcome of aging, intimately linked to the individual's health status as well as frailty.

Cedrin Mitigates Dyslipidemia, Inflammation, and Oxidative Stress in Streptozotocin-induced Diabetes Mellitus in Rats

Background Cedrin is a major bioactive flavonoid compound present in the Cedrus deodara plant and has been reported to have several biological activities. Diabetes mellitus is an endocrine and metabolic disease marked by high blood sugar levels leading to reduced insulin production and/or decreased insulin efficacy. Hyperglycemia leads to chronic health complications and organ dysfunction, specifically affecting the nerves, blood vessels, kidneys, eyes, liver, and heart. Purpose The aim of this work is to study the effect of cedrin on glycemic control, hyperlipidemia, liver function markers, and biochemical markers of streptozotocin (STZ)-induced diabetes in rats. Methods The STZ was utilized to initiate diabetes in experimental rats, and after the onset of diabetes, the rats were treated with cedrin for 30 days. After the treatment periods, the experimental rats’ glucose levels, food and water uptake, and body and organ weights were assessed. The biochemical markers, including insulin, hexokinase, glycated hemoglobin, and so on, were investigated using diagnostic kits. The concentrations of oxidative marker levels, inflammatory cytokines, and lipid marker levels were assessed using diagnostic kits. Results Cedrin treatment effectively mitigated the glucose level, food and water consumption, organ weights, and glycated hemoglobin (HbA1c) while increasing the body weight and hexokinase levels of the STZ-induced rats. In the STZ-induced rats, the cedrin treatment remarkably elevated the antioxidant levels, reduced oxidative stress biomarkers, and decreased the inflammatory cytokine levels in their serum. Conclusion The current study demonstrates that cedrin treatment effectively regulated dyslipidemia, inflammatory response, and oxidative stress in rats with STZ-induced diabetes. Therefore, it can be a potential salutary agent to treat diabetes in the future.

MiR‐105‐5p/PTEN Axis Modulates the Immune Response and Epithelial‐Mesenchymal Transition of Colon Cancer via NF‐κB Activation

ABSTRACTThe underlying regulating mechanisms of miR‐105‐5p/PTEN in colon cancer (CC) progression are still unknown. MiR‐105‐5p and PTEN expressions were determined using RT‐PCR. PTEN protein levels were examined by western blot. Also, the contents of inflammatory cytokines (IL–1β, TNF–α, and IL‐6) were measured via ELISA. An inflammation model was constructed via LPS. NF–κB p65 expression was assessed via immunofluorescence assay. A xenograft tumor model was constructed in BALB/c nude mice. The functions of miR‐105‐5p were investigated by establishing a xenograft tumor model, H&amp;E staining, TUNEL assay, immunohistochemistry assay, ELISA, RT‐PCR, and western blot. In this research, We found that miR‐105‐5p expressions were upregulated in CC cells. MiR‐105‐5p depletion notably augmented PTEN expressions and enhanced immune response, while impeded EMT and distinctly declined the levels of p‐IκBα and Nuc‐NF‐κB p65 in LoVo cells. Whereas, these effects were notably counteracted by PTEN depletion. MiR‐105‐5p upregulation exerted the opposite effects in CaCo2 cells. LPS markedly increased miR‐105‐5p expressions and suppressed PTEN expressions in LoVo cells. MiR‐105‐5p depletion offset LPS‐triggered promoting effects on EMT and suppressing effects on the immune response. Meanwhile, in vivo assay proved that miR‐105‐5p depletion markedly impeded tumor growth and EMT, yet facilitated apoptosis and immune response. It also distinctly deactivated the NF‐κB pathway. To sum up, these data indicated that miR‐105‐5p depletion might impede EMT, yet enhance the immune response in CC by elevating PTEN expressions via deactivation of the NF‐κB pathway.

Exploring the mechanism of Phyllanthus urinaria L. against ulcerative colitis based on network pharmacology and in vivo experiments

ObjectiveThis study aims to explore the therapeutic effect and molecular mechanism of the aqueous extract of Phyllanthus urinaria L. (PUL) on ulcerative colitis (UC). MethodPUL was administered to UC mice induced by 2.25% DSS. The changes of body weight, DAI score and colon length were recorded during the experiment. HE staining was conducted for pathology analysis of the mice in each group. The contents of inflammatory cytokines in colon tissues were detected by Elisa. Additionally, UPLC-TOF-MS/MS method was employed to identify the main components of PUL. Then, the TCMSP database was utilized to explore the potential active ingredients and drug targets of PUL. The matching of drug targets and disease targets yielded cross targets, which were used to construct a PPI network in String. Key targets underwent GO and KEGG enrichment analysis. Finally, Western blotting was performed to verify the key proteins expressions in the predicted pathway of network pharmacology and the expressions of tight junction proteins in the colon tissue. ResultPUL has been found to effectively alleviate the symptoms such as weight loss, bloody stools, and colon shortening in mice with UC. Administration of PUL led to an increase in tight junction protein in the colonic tissue of mice, as compared to the model group. Elisa results revealed that PUL reduced the expression of pro-inflammatory factors in mice with UC. HE staining results show that PUL can alleviate colon tissue damage caused by UC. A total of 773 components were detected in the water extract of PUL, among which 26 components, including quercetin, epigallocatechin gallate and kaempferol, may possess anti-UC activity. Network pharmacology analysis suggested that PUL may play an anti-ulcerative colitis role by inhibiting TNF pathway. Finally, Western blotting results confirmed that the PUL inhibited the TNF pathway and effectively treated ulcerative colitis. ConclusionThese preliminary research results indicate that PUL has the potential to exhibit anti-inflammatory activity by inhibiting the TNF pathway, thereby alleviating symptoms associated with UC. Substances such as quercetin, epigallocatechin gallate and kaempferol in PUL are believed to play an important role in this process.

FABP4 Inhibitor Alleviates Lipopolysaccharide-Induced HUVEC Injury by Inactivating NF-κB and Activating PPARγ.

The study aims to examine the effect of FABP4 on inflammatory response and angiogenesis in the cell model of atherosclerosis and to explore its potential mechanism.Using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting, the mRNA and protein levels of FABP4 in human umbilical vein endothelial cells (HUVECs) treated with lipopolysaccharide (LPS) or oxidized low-density lipoprotein for 6, 12, 24, or 48 hours were measured. To silence FABP4 expression and NF-κB signaling in HUVECs, FABP4 inhibitor and NF-κB inhibitor were utilized. To assess cell survival rate and apoptosis, methyl thiazolyl tetrazolium assays and flow cytometry analysis were conducted. Genes related to cholesterol metabolism, including LOX-1, ABCA1, and ABCG1, were subjected to RT-qPCR and western blotting. Moreover, protein levels of apoptotic markers (Bcl-2 and Bax), PPARγ, the phosphorylated levels of NF-κB p65, and angiogenetic markers (ICAM1, VCAM1, and VEGF) were quantified via western blotting. Using an enzyme-linked immunosorbent assay, concentrations of inflammatory cytokines in HUVECs were measured.FABP4 expression was upregulated in LPS-stimulated HUVECs. The silencing of FABP4 lowered LOX-1 and p65 levels while upregulating ABCA1 and ABCG1 expression in the context of LPS. Furthermore, the inhibition of FABP4 or NF-κB signaling promoted the growth of HUVECs, upregulated Bcl-2 and PPARγ protein levels, and reduced Bax levels, angiogenetic markers, and inflammatory cytokines in the context of LPS. The combination of FABP4 inhibitor and NF-κB inhibitor treatment amplified the above-mentioned effects on cell growth, angiogenesis, and inflammation.Inhibition of FABP4 reduces LPS-induced HUVEC cell damage via the inactivation of NF-κB p65 and activation of PPARγ signaling.

The links between neuroinflammation, brain structure and depressive disorder: A cross-sectional study protocol.

It is known that symptoms of major depressive disorder (MDD) are associated with neurodegeneration, that lipopolysaccharide (LPS) can induce symptoms of MDD, and that blood LPS levels are elevated in neurodegeneration. However, it is not known whether blood LPS and cytokine levels correlate with MDD, cognition and brain structure, and this is tested in this study. This cross-sectional study includes individuals with MDD (n = 100) and a control group of individuals with no one-year history of a mental disorder (n = 50). A comprehensive evaluation is performed, including the collection of basic sociodemographic information, data on smoking status, body mass index, course of MDD, past treatment, comorbid diseases, and current use of medications. Diagnosis of MDD is performed according to the WHO's [2019] International Classification of Diseases and related health problems by psychiatrist and severity of MDD is evaluated using the Montgomery-Åsberg Depression Scale. The Cambridge Neuropsychological Test Automated Battery is used to evaluate cognitive functioning. Venous blood samples are taken to measure genetic and inflammatory markers, and multiparametric brain magnetic resonance imaging is performed to evaluate for blood-brain barrier permeability, structural and neurometabolic brain changes. Descriptive and inferential statistics, including linear and logistic regression, will be used to analyse relationships between blood plasma LPS and inflammatory cytokine concentrations in MDD patients and controls. The proposed sample sizes are suitable for identifying significant differences between the groups, according to a power analysis. Trial registration: Clinicaltrials.gov NCT06203015.

Concomitant parasite infections influence tuberculosis immunopathology and favor rapid sputum conversion of pulmonary tuberculosis patients

Immunopathology of human tuberculosis (TB) in a subgroup of patients is characterized by aberrantly high concentrations of inflammatory cytokines, for example Interleukin (IL)-6. Concomitant (co-)infections by parasites can affect host immunity, but the impact on immunopathology in TB patients is poorly defined. Here we characterized a group of patients with TB ( n = 76) from Ghana with different protozoan and helminth co-infections. Plasma cytokines were measured at the onset of disease and anti-mycobacterial treatment efficacy was monitored during disease course. A subgroup of TB patients had co-infections with protozoan (n = 19) or helminth (n = 16) parasites. Plasma analyses for candidate cytokines identified lower levels of IL-6 in parasite co-infected patients with TB. Moreover, it took less time for co-infected patients to become sputum-negative for Mycobacterium tuberculosis during treatment. These results indicated an influence of parasite co-infections on immunopathology in TB and suggested positive effects on treatment efficacy.

Hydroxy-selenomethionine helps cows to overcome heat stress by enhancing antioxidant capacity and alleviating blood-milk barrier damage

Heat stress can lead to decreased feed intake, apoptosis of mammary epithelial cells, and decreased milk yield and quality. Selenium is an important element in the composition of at least 25 selenoproteins. Hydroxy-selenomethionine (HMSeBA; Andy Su Life Science Products Co., Ltd., Shanghai, China) is a novel organic selenium that has been shown to have a better deposition effect. However, whether HMSeBA alleviates damage to the mammary gland blood-milk barrier caused by heat stress and how this affects the performance of dairy cows remain largely unexplored. Therefore, 32 healthy Holstein cows with similar gestation days (150.41 ± 20.07 d), milk yield (36.15 ± 3.02 kg) and parity (3.25 ± 0.51) were selected and randomly divided into two total mixed rations with different selenium (Se) sources: sodium selenite (SSe) and HMSeBA. This study evaluated the outcomes of HMSeBA on antioxidant capacity, immunity, and blood-milk barrier damage in dairy cows during heat stress by collecting the samples of blood, rumen fluid and mammary gland biopsy. The experiment was conducted over 35 d, including a 5-day pre-feeding period and a 30-day experimental period. The temperature and humidity index (THI) were all above 80 throughout the experiment period. The results showed that HMSeBA decreased the respiratory rate (P < 0.001) and the content of inflammatory cytokines in the serum and increased the content of immune factors and antioxidant capacity (P < 0.05). In addition, HMSeBA reduced the expression of inflammatory cytokines and heat shock proteins in mammary gland (P < 0.05). Hematoxylin-eosin-stained pathological sections showed massive thickening of acinar walls and severe destruction of glandular structures in the SSe group, but the structure of the acinar mammary gland in the HMSeBA group was intact. Furthermore, HMSeBA promoted the expression of the phosphatidylinositol 3-kinase (PI3K, P < 0.001)/protein kinase B (AKT, P = 0.011)/mammalian target of rapamycin (mTOR, P = 0.008) pathway and improved the expression of zonula occludens-1 (ZO-1, P = 0.014) and occluding (OCLN, P = 0.012) in the mammary gland, suggesting less damage caused by heat stress to the blood-milk barrier. Our results demonstrated that HMSeBA can improve the antioxidant capacity and immunity of dairy cows and the expression of tight junction proteins in mammary gland to help alleviate the blood-milk barrier damage by heat stress, which could reduce the damage of heat stress on milk yield.

Harmony in Healing: Investigating Platelet-Rich Plasma Activation during Acetylsalicylic Acid Treatment

Platelet-rich plasma (PRP) therapy holds promise for treating various clinical conditions. The activation process is crucial in releasing growth factors and cytokines from platelets, enhancing the therapeutic properties of PRP. Standard activation methods involve autologous thrombin or collagen, with variations in efficacy and growth factor release. This study explores the impact of acetylsalicylic acid (ASA), a commonly used antiplatelet drug, on PRP activation. The results indicate that non-activated PRP extracted from the whole blood of ASA-treated patients exhibits increased inflammatory cytokine concentrations, notably TNFa. After activation with autologous thrombin/CaCl2 or collagen IV, the measured fluorescence intensities suggest varying release patterns between treated and non-treated groups. Understanding the influence of ASA on platelet activation holds implications for personalized medicine and optimizing outcomes for individual patients undergoing PRP therapy. This research sheds light on the potential challenges associated with using antiplatelet drugs, emphasizing the need for careful consideration in tailoring PRP-based regenerative therapies.

Formononetin alleviates ulcerative colitis via reshaping the balance of M1/M2 macrophage polarization in a gut microbiota-dependent manner

BackgroundUlcerative colitis (UC), a type of inflammatory bowel disease, presents substantial challenges in clinical treatment due to the limitations of current medications. Formononetin (FN), a naturally compound with widespread availability, exhibits anti-inflammatory, antioxidant, and immunomodulatory properties. PurposeThis study aimed to investigate the efficacy of FN against UC and its potential regulatory mechanism. MethodsHere, dextran sulfate sodium (DSS) was employed to replicate experimental colitis in mice with concomitant FN treatment. The distribution and localisation of CD68 and F4/80 macrophages in colonic tissues were visualized by immunofluorescence, their chemokine and inflammatory cytokine concentrations were determined by ELISA, and macrophages and M1/M2 subpopulations were determined by flow cytometry. Additionally, 16 s rRNA and LC-MS techniques were used to detect the colonic intestinal microbiota and metabolite profiles, respectively. Correlation analyses was performed to clarify the interactions between differential bacteria, metabolites and M1/M2 macrophages, and pseudo sterile mice were constructed by depletion of gut flora with quadruple antibiotics, followed by faecal microbial transplantation to evaluate its effects on colitis and M1/M2 macrophage polarisation. ResultsFN dose-dependently alleviated clinical symptoms and inflammatory injury in colonic tissues of colitis mice, with its high-dose efficacy comparable to that of 5-ASA. Concurrently, FN not only inhibited inflammatory infiltration of macrophages and their M1/M2 polarisation balance in colitis mice, but also improved the composition of colonic microbiota and metabolite profiles. However, FN lost its protective effects against DSS-induced colitis and failed to restore the equilibrium of M1/M2 macrophage differentiation following intestinal flora depletion through quadruple antibiotic treatment. Importantly, fecal microbiota transplantation from FN-treated mice restored FN's protective effects against DSS-induced colitis and reestablished its regulatory role in M1/M2 macrophage polarization. ConclusionCollectively, FN ameliorated UC through modulating the balance of M1/M2 macrophage polarization in a gut microbiota-dependent manner.

Cimifugin Alleviates Chronic Constriction Injury of the Sciatic Nerve by Suppressing Inflammatory Response and Schwann Cell Apoptosis.

Inflammation and Schwann cell apoptosis play critical roles in neuropathic pain after sciatic nerve injury. This study aimed to explore the function and mechanism of cimifugin in lipopolysaccharide (LPS)-stimulated rat Schwann cells and sciatic nerves of rats treated with chronic constriction injury (CCI). Thermal, mechanical and cold hyperalgesia of rats in response to cimifugin or mecobalamin (the positive drug control) treatment were evaluated through behavioral tests. H&E staining of sciatic nerves was performed for pathological observation. ELISA was conducted to assess concentrations of inflammatory cytokines in rat serum and sciatic nerves. The intensity of S100β in sciatic nerves was determined using immunohistochemistry. Flow cytometry analysis was conducted for detection of Schwann cell apoptosis. RT-qPCR was performed to measure mRNA levels of inflammatory factors in Schwann cells. Immunofluorescence staining was performed to detect cellular p65/NF-κB activity. Western blotting was performed to quantify protein levels of apoptotic markers and factors associated with the NF-κB and MAPK pathways in rat nerves and Schwann cells. As shown by experimental data, cimifugin mitigated thermal, mechanical and cold hyperalgesia of CCI rats. Cimifugin repressed inflammatory cell infiltration, reduced proinflammatory cytokine levels while increasing anti-inflammatory factor (IL-10) level in serum or sciatic nerves of CCI rats. Cimifugin enhanced S100β expression and downregulated apoptotic markers in vivo. The anti-inflammatory and anti-apoptotic properties of cimifugin were verified in the LPS-stimulated Schwann cells. Moreover, cimifugin suppressed nuclear translocation of p65 NF-κB in vitro and repressed the phosphorylation of IκB, p65 NF-κB, p38 MAPK, ERK1/2, as well as JNK in CCI rats. In conclusion, cimifugin alleviates neuropathic pain after sciatica by suppressing inflammatory response and Schwann cell apoptosis via inactivation of NF-κB and MAPK pathways.

Nasal Mucosal Cytokines as Potential Biomarkers for Assessing Disease Severity and Class of Pathogen in Children with Community-Acquired Pneumonia.

Community-acquired pneumonia (CAP) is a leading cause of morbidity and mortality in children. Assessing disease severity and etiology remains challenging in the clinical setting. The objective of this study was to identify mucosal biomarkers that could potentially assist with patient classification. We analyzed mucosal concentrations of cytokines in nasopharyngeal samples obtained from a convenience sample of 182 children with CAP and 26 matched healthy controls. Pathogens were identified by cultures and molecular assays. Severe disease was defined by hospital stay ≥ 3 days, and/or PICU admission. Data were analyzed according to identified pathogens and disease severity. Children with CAP and detected atypical bacteria had significantly higher concentrations of MCP-2, IFN-γ and CXCL10 among others compared with those with typical bacteria. Children with influenza virus had significantly higher concentrations of MCP-2, CXCL10, CXCL11, CX3CL1, and IFN-γ than those with typical bacteria. Additionally, children with severe CAP had significantly higher concentrations of CCL23 than children with mild/moderate disease, irrespective of the pathogen(s) identified. We identified differences in mucosal concentrations of inflammatory and antiviral cytokines in children with CAP according to disease severity and detected pathogens. Mucosal biomarkers represent a promising approach to help assessing disease severity and etiology.

Approaches to Identifying Markers of Effect of Environment and Nutrition in Preschoolers

Background: Identification of the markers of effect of environment and nutrition on the human body, given their interplay, remains relevant for health risk assessment. Objective: To identify markers of children’s health effects of environmental exposures and nutrition with account for their mutual influences. Materials and methods: The study involved 197 preschool children living in two towns of the Sverdlovsk Region with different levels of environmental pollution. In January 2022 to June 2024, we tested mass concentrations of 19 metals, cytogenetic parameters of buccal epithelium, interleukins (IL-1, IL-4), glutathione-S-transferase, and 60 organic acids in urine of the subjects. We also evaluated the preschool menu and off-school nutrition, anthropometric parameters, morbidity based on outpatient cards, and health status using a questionnaire-based survey. Microsoft Excel and IBM SPSS were used for statistical data analysis. Results: We established that in the children living in the territory with high airborne risks, the levels of exposure to seven chemicals exceeded the reference values, especially those of aluminum and manganese by 3.9 and 2.5 times, respectively. Cytogenetic damage to buccal epithelial cells, higher values of markers of allergic reactions (IL 4), higher values of glutathione-S-transfer and organic acids, and markers of detoxification were also more frequent in this group (p &lt; 0.001). We noted that the diet of children was deficient in vitamins B1 and C, calcium, and, judging by the markers of organic acids, essential amino acids and vitamins B9 and B12. The exposed children differed from the controls in high exposure to arsenic, mercury, and copper; there was also a larger proportion (85 %) of children with cells with micronuclei and atypical nuclei, high concentrations of inflammatory cytokines (IL 1), energy and protein deficiencies, excess sugars in the diet, and markers of energy metabolism disorders among them. Conclusions: The identified markers of two non-adaptive metabolotypes to environmental exposure and nutritional phenotypes will allow a differentiated approach to developing diets.

Changes in Type 1 Diabetes-Associated Gut Microbiota Aggravate Brain Ischemia Injury by Affecting Microglial Polarization Via the Butyrate-MyD88 Pathway in Mice.

People with type 1 diabetes (T1D) have a significantly elevated risk of stroke, but the mechanism through which T1D worsens ischemic stroke remains unclear. This study was aimed at investigating the roles of T1D-associated changes in the gut microbiota in aggravating ischemic stroke and the underlying mechanism. Fecal 16SrRNA sequencing indicated that T1D mice and mice with transplantation of T1D mouse gut microbiota had lower relative abundance of butyric acid producers, f_Erysipelotrichaceae and g_Allobaculum, and lower content of butyric acid in feces. After middle cerebral artery occlusion (MCAO), these mice had poorer neurological outcomes and more severe inflammation, but higher expression of myeloid differentiation factor 88 (MyD88) in the ischemic penumbra; moreover, the microglia were inclined to polarize toward the pro-inflammatory type. Administration of butyrate to T1D mice in the drinking water alleviated the neurological damage after MCAO. Butyrate influenced the response and polarization of BV2 and decreased the production of inflammatory cytokines via MyD88 after oxygen-glucose deprivation/reoxygenation. Knocking down MyD88 in the brain alleviated neurological outcomes and decreased the concentrations of inflammatory cytokines in the brain after stroke in mice with transplantation of T1D mouse gut microbiota. Poor neurological outcomes and aggravated inflammatory responses of T1D mice after ischemic stroke may be partly due to differences in microglial polarization mediated by the gut microbiota-butyrate-MyD88 pathway. These findings provide new ideas and potential intervention targets for alleviating neurological damage after ischemic stroke in T1D.

Remimazolam ameliorates postoperative cognitive dysfunction after deep hypothermic circulatory arrest through HMGB1-TLR4-NF-κB pathway

BackgroundPostoperative cognitive dysfunction (POCD) is a complication of deep hypothermic circulatory arrest (DHCA). Various amounts of neurologic dysfunctions have been shown after DHCA, which has often been attributed to systemic inflammatory response syndrome and cerebral ischemia/reperfusion injury. Remimazolam is one of the commonly used anesthetic drugs with protective actions against inflammatory diseases, such as sepsis and cerebral ischemia/reperfusion injury. Here, we determined the protective effect and potential mechanism of action of remimazolam against neuronal damage after DHCA. MethodsA rat model of DHCA was established, and a gradient dosage of remimazolam was administered during cardiopulmonary bypass (CPB). The cognitive function of rats was evaluated by Morris water maze. Hematoxylin and eosin and TUNEL staining were performed to assess hippocampus tissue injury and neuronal apoptosis. Inflammatory cytokines concentration were analyzed by enzyme-linked immunosorbent assay. The protein expression was analyzed using automated electrophoresis western analysis and immunohistochemical analysis. ResultsThe appropriate dosage of remimazolam reduced histologic injury, neuronal apoptosis, microglia activation, and secondary inflammatory cascades, as well as the downregulation of the expression of the HMGB1-TLR4-NF-κB pathway after DHCA, improved the memory and learning abilities in DHCA rats. Further, administration of a TLR4 antagonist TAK-242 had a similar effect to remimazolam, while the TLR4 agonist LPS attenuated the effect of remimazolam. ConclusionsRemimazolam could ameliorate POCD after DHCA through the HMGB1-TLR4-NF-κB signaling pathway.

Zonulin — effective biomarker for the diagnosis of intestinal lesions in patients with nonalcoholic fatty liver disease in COVID‑19

The SARS‑CoV‑2 virus mainly attacks the respiratory system, but can also affect several organs, in particular, causing gastrointestinal symptoms such as vomiting, diarrhea or abdominal pain in the early stages of the disease. Intestinal dysfunction leads to a change in intestinal microorganisms and an increase in the content of inflammatory cytokines. Liver dysfunction is also relatively common in patients with coronavirus disease 2019 (COVID19). Objective — to investigate the efficacy of zonulin in determining intestinal damage in patients with nonalcoholic fatty liver disease with COVID‑19. Materials and methods. The study included 76 patients with non‑alcoholic fatty liver disease (NAFLD) who had complaints indicating intestinal disorders during the outpatient stage of follow‑up after ­COVID‑19. From the anamnesis, it was found that all examined patients had a confirmed diagnosis of infection with SARS‑CoV‑2 virus. All patients had a mostly mild course of COVID‑19, which did not require hospitalization. Patients with NAFLD were divided into two groups depending on defecation violation: group I — 36 patients who, after an acute respiratory infection caused by the SARS‑CoV‑2 virus, developed periodic diarrhea (on average, in (38.7±5.2) days after a negative PCR result for SARS‑CoV‑2 virus); group II — 40 patients with constipation, which began to bother on average in (86.9±7.4) days after a negative PCR result. The control group included 20 practically healthy individuals. The level of zonulin was determined in blood serum and faeces by enzyme‑linked immunosorbent assay. Patients underwent faecal culture to assess the quantitative and qualitative composition of the colon microflora. Results. During the anthropometric examination, overweight and first‑degree obesity were detected more frequently in patients of both groups: overweight was diagnosed most frequently in patients of group I (38.9%) and first‑degree obesity in patients of group II (35.0%). The microbiological examination of faeces in both groups of patients revealed a decrease in the number of bifidus and lactobacilli, as well as Enterococcus and E. coli with normal enzymatic properties. In group I patients, Citrobacter and Staphylococcus were statistically significantly (p &lt;0.05) more often detected in increased concentrations in faeces, whereas in group II patients Enterobacter, Klebsiella and Clostridium were detected (p &lt;0.05). Significant changes were found in the analysis of zonulin levels in both serum and faeces in patients with NAFLD after COVID‑19. Serum zonulin levels in patients of group I increased 6.5‑fold, and in patients of group II—8.6‑fold (p &lt;0.001). An increase in the concentration of zonulin in the faeces was also detected — by 7.5 and 9.7 times, respectively (p &lt;0.001). A correlation was established between the level of zonulin in both blood serum and faeces and a decrease in the number of bifidus and lactobacilli in faeces in patients of both groups. Also, a strong correlation was found between the number of Enterobacter, Klebsiella and Clostridium and a medium intensity correlation between the number of Staphylococcus in the faeces in patients of group ІІ. In patients of group І, a correlation was found between the number of Staphylococcus and Citrobacter. The level of zonulin (both in blood serum and faeces) directly depended on changes in body mass index in the patients we examined, namely, overweight in patients of group І and obesity of the first degree in patients of group ІІ. Conclusions. In patients with NAFLD after COVID19, changes in the quantitative and qualitative composition of the colon microflora were found in the intestinal lesions, including a decrease in the number of bifidobacteria and lactobacilli, as well as an increase in the content of Staphylococcus and Citrobacter in the faeces in patients with predominant diarrhoea and Enterobacter, Klebsiella and Clostridium in patients with predominant constipation. An increase in the level of zonulin in the blood serum and faeces in patients with NAFLD after ­COVID‑19 was detected. A more pronounced deviation from the norm was found in patients of group ІІ (with complaints of constipation). A direct correlation was established between the level of zonulin in the blood serum and faeces and changes in the quantitative and qualitative composition of the colon microflora, as well as an increase in body mass index in patients with NAFLD after ­COVID‑19.

PSI-8 Inflammatory cytokine concentrations and gene expression in mature and aging sedentary horses

Abstract Chronic inflammation associated with aging, termed “inflamm-aging” may negatively impact overall animal health. However, the mechanisms of inflamm-aging and, therefore, effective interventions, remain unknown. Nuclear factor erythroid 2-related factor 2 (Nrf2; NFE2L2) is a transcription factor that controls antioxidant gene expression and could be a target for promoting anti-inflammatory pathways. To test the hypothesis that supplementation of nonenzymatic derived antioxidants (Protandim Nrf2 Synergizer, LifeVantage Corporation) would activate Nrf2 and mitigate inflammation, 40 mature, sedentary horses [32 mares, 8 geldings; 15.7 ± 4.9 yr of age, body weight (BW) = 519 ± 46 kg] were stratified by age, sex, and BW and randomly assigned to one of four dietary treatment groups for 56 d: 1) 0 mg (CON); 2) 675 mg (Pro1); 3) 2,025 mg (Pro3); or 4) 4,050 mg (Pro6) Protandim Nrf2 Synergizer per day (n = 10 per group). Blood samples were collected at d 0 (September, 27°C, 61.6% humidity), d 28 (October, 21.6°C, 55.8% humidity), and d 56 (November, 15.1°C, 73.8% humidity) of supplementation prior to the morning feeding. Plasma cytokine concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-4, IL-6, IL-8, and IL-10 were quantified using custom Meso Scale Discovery U-Plex kits and whole blood gene expression of IL-1β, NFE2L2, and heme oxygenase 1 (HMOX1) were determined using qRT-PCR. Data were analyzed using PROC MIXED in SAS v9.4 with sex, time, treatment, age (under or over 15 yr), and time × treatment as fixed effects and time as a repeated effect with horse(treatment) as the subject. Day 0 was included as a covariate when treatment groups differed at d 0. Relationships between age and cytokine concentrations or mRNA expression were determined using PROC CORR. Plasma IL-4 (r = 0.4054) concentrations as well as whole blood HMOX1 (r = 0.3437) and NFE2L2 (r = 0.3382) expression on d 0 were positively correlated with age (P ≤ 0.04). Regardless of treatment or age, IL-1β and NFE2L2 expression and IL-4 and IL-8 concentrations decreased from d 0 to 28 (P ≤ 0.03) and remained depressed at d 56 (P ≤ 0.02) while IL-10 was not different from d 0 to 28 but decreased from d 0 to 56 (P = 0.02). TNFα concentrations were not different between treatments or over time. These data corroborate increased inflammatory markers with age and suggest a change in inflammatory status over time potentially due to seasonal environmental factors.