Anti-inflammatory Cytokine Genes Research Articles

Isolation and characterization of plant-derived exosome-like nanoparticles from Carica papaya L. fruit and their potential as anti-inflammatory agent.

Inflammation is an immune system response that identifies and eliminates foreign material. However, excessive and persistent inflammation could disrupt the healing process. Plant-derived exosome-like nanoparticles (PDENs) are a promising candidate for therapeutic application because they are safe, biodegradable and biocompatible. In this study, papaya PDENs were isolated by a PEG6000-based method and characterized by dynamic light scattering (DLS), transmission Electron Microscopy (TEM), bicinchoninic acid (BCA) assay method, GC-MS analysis, total phenolic content (TPC) analysis, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. For the in vitro test, we conducted internalization analysis, toxicity assessment, determination of nitrite concentration, and assessed the expression of inflammatory cytokine genes using qRT-PCR in RAW 264.7 cells. For the in vivo test, inflammation was induced by caudal fin amputation followed by analysis of macrophage and neutrophil migration in zebrafish (Danio rerio) larvae. The result showed that papaya PDENs can be well isolated using the optimized differential centrifugation method with the addition of 30 ppm pectolyase, 15% PEG, and 0.2 M NaCl, which exhibited cup-shaped and spherical morphological structure with an average diameter of 168.8±9.62 nm. The papaya PDENs storage is stable in aquabidest and 25 mM trehalose solution at -20˚C until the fourth week. TPC estimation of all papaya PDENs ages did not show a significant change, while the DPPH test exhibited a significant change in the second week. The major compounds contained in Papaya PDENs is 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP). Papaya PDENs can be internalized and is non-cytotoxic to RAW 264.7 cells. Moreover, LPS-induced RAW 264.7 cells treated with papaya PDENs showed a decrease in NO production and downregulation mRNA expression of pro-inflammatory cytokine genes (IL-1B and IL-6) and an upregulation in mRNA expression of anti-inflammatory cytokine gene (IL-10). In addition, in vivo tests conducted on zebrafish treated with PDENs papaya showed inhibition of macrophage and neutrophil cell migration. These findings suggest that PDENs papaya possesses anti-inflammatory properties.

Proinflammatory cytokines IL-6, IL-1β, TNF-α in infective endocarditis

Aim. To study the features of macrophages in the tissues of resected valves in operated patients with infective endocarditis (IE), their significance and interaction with inflammatory markers to improve the effectiveness of IE diagnosis. Materials and methods. Prospectively the research included 25 adult patients with active IE (Duke criteria 2015) and 24 patients with heart defects without IE, hospitalized in a cardiosurgical hospital in Moscow (2021–2022). A standard laboratory and instrumental examination was carried out for the diagnosis of IE, including etiological diagnosis with microbiological and molecular biological methods, and echocardiographic examination of heart. Additionally, the neutrophil-to-lymphocyte ratio (NLR) was calculated. The study of macrophages was carried out in the tissues of resected valves with the determination of the expression of pro- and anti-inflammatory cytokine genes, macrophage markers (CD 68+) using real-time PCR. Results. Increased expression of proinflammatory cytokines IL-1β, TNF-α and IL-6 was revealed in the group of operated patients with IE with significant differences in IL-1β (CI [IQR] 0.00367 [0.00047–0.01553] vs 0.00018 [0.00012–0.00262]; p0.05) and IL-6 (CI [IQR] 0.00367 [0.00047–0.01553] vs 0.00018 [0.00012–0.00262]; p0.05) and IL-6 (CI [IQR] 0.00338 [0.00066–0.01674] vs 0.00054 [0.00044–0.00378]; p0.05). The expression of anti-inflammatory cytokines in valve tissues prevailed in the control group without significant differences from patients with IE. The macrophage marker CD 68+ was revealed in all examined patients with a significant quantitative predominance in the group of patients with IE. There were no differences in the expression of pro- and anti-inflammatory cytokines depending on the presence of embolic events, intracardiac complications, etiological affiliation to S. aureus, as well as hospital mortality and combined endpoint (death from all causes or recurrence of IE 6 months after surgery) in patients with IE with or without events. Cytokines IL-1β and IL-6 positively correlated with each other, with leukocytes and NLR. ROC analysis determined that IL-1β and NLR had the most favorable features for the diagnosis of IE [IL-1β AUC 0.816 (p=0.02), NLR AUC 0.807 (p=0.03)]. IL-6 did not show a diagnostic value in IE. The threshold value for IL-1β was 0.00029 (sensitivity 86.4%, specificity 60.0%, prognostic value of negative result 75.0% and positive 76.0%, AUC 0.761; p=0.008). Conclusion. The valve macrophages of patients with IE express elevated levels of proinflammatory cytokines IL-1β and IL-6, regardless of etiological affiliation or complicated course of IE. IL-1β has a high diagnostic value for determining the inflammatory activity in IE.

Polymorphism of IL4 and IL13 anti-inflammatory cytokine genes in indigenous populations of the russian Arctic

Living conditions in the Arctic zone are extremely unfavorable for living and contribute to the development of a number of diseases due to suppression of the functions of the immune system. Cytokines are one of the main mediators of the immune system; they are encoded by genes with a high degree of polymorphism. This study examined the distribution of genetic variants in the cytokine genes (rs2243250 IL4, rs1800925 IL13) among the Nenets, Dolgan-Nganasan and Slavs. It was previously established that these mutations are associated with the level of expression of these interleukins and their production, which leads to changes in the impairment of the immune response to the influence of the pathogen. The study showed the predominance of genotypes CT and TT of the rs2243250 IL4 polymorphism, and genotype CC of the rs1800925 IL13 polymorphism in the Nenets and Dolgan-Nganasan populations. The results obtained suggest that the indigenous inhabitants of the Russian Arctic have a genetically determined rapid immune response and protection against the development of allergic diseases compared to the Slavs.

Enterococcus durans 98D alters gut microbial composition and function to improve DSS-induced colitis in mice

Enterococcus durans, is a potential functional strain with the capacity to regulate intestinal health and ameliorate colonic inflammation. However, the strain requires further investigation regarding its safety profile and potential mechanisms of colitis improvement. In this study, the safety of E. durans 98D (Ed) as a potential probiotic was studied using in vitro methods. Additionally, a dextran sulfate sodium (DSS)-induced murine colitis model was employed to investigate its impact on the intestinal microbiota and colitis. In vitro antimicrobial assays revealed Ed sensitivity to common antibiotics and its inhibitory effect on the growth of Escherichia coli O157, Streptococcus pneumoniae CCUG 37328, and Staphylococcus aureus ATCC 25923. To elucidate the functional properties of Ed, 24 weight-matched 6-week-old female C57BL/6J mice were randomly divided into three groups (n = 8): NC group, Con group (DSS), and Ed group (DSS + Ed). Ed administration demonstrated a protective effect on colitis mice, as evidenced by improvements in body weight, colonic length, reduced disease activity index, histological scores, diminished splenomegaly, and decreased goblet cell loss. Furthermore, Ed downregulated the expression of the pro-inflammatory cytokine genes (IL-6, IL-1β, and TNF-α) and upregulated the expression of the anti-inflammatory cytokine gene IL-10. The 16S rRNA gene sequencing revealed significant alterations in microbial α-diversity, with principal coordinate analysis indicating distinct differences in microbial composition among the three groups. At the phylum level, the relative abundance of Actinomycetota significantly increased in the Ed-treated group. At the genus level, Ed treatment markedly elevated the relative abundance of Paraprevotella, Rikenellaceae_RC9, and Odoribacter in DSS-induced colitis mice. In conclusion, Ed exhibits potential as a safe and effective therapeutic agent for DSS-induced colitis by reshaping the colonic microbiota.

Effects of extended dietary supplementation with Santalum album essential oil on hemato-biochemical changes, innate immune response, antioxidant status, and expression of related gene in Nile tilapia (Oreochromis niloticus).

The effects of long-term dietary supplementation with sandalwood (Santalum album L.) essential oil (SEO) was investigated on hemato-biochemical biomarkers, immune status, antioxidant capacity, and resistance against Staphylococcus aureus in Nile tilapia (Oreochromis niloticus). Five groups (with four replicates) of O. niloticus (12.60 ± 0.20 g) were fed diets supplemented with SEO at doses of 0, 0.5, 1.0, 2.0, and 4.0 mL/kg diet for 60 days. Results indicated a substantial increase in blood protein levels and lower serum cholesterol, cortisol, glucose, urea, creatinine levels and, transaminase activities of fish fed a 2.0-mL SEO/kg diet. Serum lysozyme activity, nitric oxide, complement-3 levels, and phagocytic activity were significantly improved in O. niloticus after 60 days of feeding SEO-supplemented diets. Dietary SEO at level of 2.0-mL SEO/kg diet increased the activities of SOD, CAT, and GPx, and decreased MDA levels in liver homogenate. In addition, dietary 2.0-mL SEO/kg diet significantly upregulated antioxidant genes expression (CAT, SOD, GPx, GST, and GSR) with downregulation of apoptotic genes (HSP70, TLR2, caspase-3, and PCNA) in the liver. Furthermore, SEO-enriched diets significantly down-regulated pro-inflammatory (TNF-α, IL-1β, and IL-8) and up-regulated anti-inflammatory cytokine genes (TFG-β and IL-10) in the spleen. Moreover, SEO fortification increased the relative percentage of survival against S. aureus challenge and regulated immune-antioxidant genes in the spleen after the challenge. Overall, the results revealed that long-term using SEO might strengthen the physiological performance, hepatic oxidant/antioxidant balance, innate immune response, and resistance of O. niloticus against bacterial infections.

Chronic social defeat stress and glucocorticoid regulation in brain regions: resistance or hypersensitivity?

Chronic social defeat stress and glucocorticoid regulation in brain regions: resistance or hypersensitivity?

The ameliorative role of methionine in hepatic steatosis and stress response in juvenile black seabream (Acanthopagrus schlegelii) fed with a high-fat diet

Methionine (Met) is not only an essential amino acid of aquatic animals, but it also regulates lipid metabolism. To investigate the impacts of dietary Met supplementation on lipid metabolism, oxidative stress (OS), inflammation and apoptosis in marine fish, a feeding trial lasting for eight weeks was conducted on juvenile black seabream (Acanthopagrus schlegelii) (initial weight 1.61 ± 0.01 g) fed with five different experimental diets: Control, regular fat diet (11% lipid level); HFD, high-fat diet (17% lipid level), HFD contained different contents of Met (5, 10 or 20 g/ kg) named HFD + M1, HFD + M2, and HFD + M3, respectively. Serum biochemical indices results indicated that the contents of triglyceride (TG), total cholesterol (TC), non-esterified fatty acid (NEFA) and low-density lipoprotein cholesterol (LDL-C), as well as the activity of alanine transaminase (ALT), markedly decreased in HFD + M1 group compared to those in the HFD treatment. Results of liver sections indicated that lipid droplets dramatically decreased by dietary Met supplementation than the HFD group. Similar results were reflected in transmission electron microscope (TEM) images. The mRNA-abundant levels of adenosine monophosphate-activated protein kinase α (ampkα) and silent information regulator 1 (sirt1) significantly increased by dietary HFD + M1 than those in the HFD treatment. When comparing to the HFD treatment, the levels of gene expression for lipogenesis pathway genes, specifically sterol regulatory element-binding protein-1c (srebp-1c) and fatty acid synthase (fas) were markedly improved by dietary HFD + M1. In contrast, opposite results were recorded in the mRNA expression levels of lipolysis pathway related genes (pparα, cpt1a, hsl, atgl, lpl) that were down-regulated by the HFD + M1 by promoting related lipogenesis. The transcriptional expression level of srebp-1c was significantly suppressed in HFD + M1 diet. HFD caused oxidative stress, inflammation and apoptosis in the liver reflected in increasing the reactive oxygen species (ROS) and malonaldehyde (MDA) contents, c-Jun N-terminal kinase (jnk) and nuclear factor kappa B (nf-κb) mRNA expression levels, but those physiological stresses could be alleviated by dietary Met supplementation. Specifically, the antioxidant enzyme activities and gene expression levels were notably up-regulated by HFD + M1 group than those in fish fed with HFD. Additionally, compared to the HFD treatment, dietary Met supplementation significantly decreased the transcriptional expression levels of pro-inflammation and pro apoptosis genes, but up-regulated the transcriptional expression of anti-inflammatory cytokine and anti-apoptosis gene. Together, this study elucidates that HFD supplemented with Met can ameliorate hepatic steatosis, oxidative stress, inflammation and apoptosis induced by HFD, confirming that Met can play a vital role in lowing lipid accumulation by regulating lipid metabolism in A. schlegelii fed with HFD.

Contribution of polymorphic variants of pro- and anti-inflammatory cytokine genes to the development of occupational lung dust pathology in miners

Introduction. The formation and progression of occupational pathology of the bronchopulmonary system during prolonged contact with fibrogenic dust are closely related to changes in the concentration of certain pro- and anti-inflammatory cytokines in the bloodstream. The change in cytokine expression is due to the presence of polymorphism of certain genes, which results in quantitative or structural changes in proteins that can affect the course and outcome of dust lung diseases.
 The study aims to determine the associations of single-nucleotide polymorphisms of cytokine genes with the formation of occupational dust pathology of the lungs.
 Materials and methods. Scientists examined 140 coal mine workers who had been exposed to high concentrations of coal and stone dust for a long time. We divided the study participants into 2 groups. The main group included 75 miners with a previously established diagnosis of "dust pathology of the lungs", the comparison group consisted of 65 men working in the same sanitary and hygienic conditions, but without an established occupational disease of the bronchopulmonary system. The research groups are comparable by gender, age and ethnicity. The scientists carried out genotyping by allele-specific polymerase chain reaction in real time.
 Results. Experts have identified a number of gene polymorphisms associated with the risk of developing occupational pathology of the bronchopulmonary system as a result of exposure to fibrogenic dust in Kuzbass miners. They also found that the following markers predispose to the formation of occupational lung pathology in miners: allele C and heterozygous genotype C/T of the polymorphic locus rs1800896 of the IL-10 gene, allele C of the polymorphism rs1800795 of the IL-6 gene, homozygous genotype T/T of the IL-4 gene (rs2243250). In turn, the T allele and the homozygous T/T genotype of the rs1800896 polymorphism of the IL-10 gene, as well as the G allele of the rs1800795 polymorphic locus of the IL-6 gene have a protective effect on the development of occupational lung dust pathology.
 Limitations. The conducted study has limitations in the form of a relatively small sample size and the lack of quantitative determination of the concentration of cytokines in the blood.
 Conclusion. The results of the conducted studies indicate the contribution of polymorphic variants of the genes of the studied pro- and anti-inflammatory cytokines to the formation of genetic predisposition and resistance to the development of dust pathology of the lungs of professional genesis. With the help of the obtained informative markers, it is possible to assess the risk of developing dust lung diseases in miners working in contact with high concentrations of coal-rock dust.
 Ethics. The Bioethical Committee of the Research Institute of Complex Problems of Hygiene and Occupational Diseases has approved the study. The examination of patients was carried out on the basis of ethical standards developed in accordance with the Helsinki Declaration of the World Medical Association "Ethical principles of conducting Scientific medical research with human participation" as amended in 2013 and "Rules of Clinical Practice in the Russian Federation" approved by Order of the Ministry of Health of the Russian Federation No. 266 dated 06/19/2003. The study was conducted with the written consent of the subjects.

The Effects of Zinc Oxide Nanoparticles on Antioxidation, Inflammation, Tight Junction Integrity, and Apoptosis in Heat-Stressed Bovine Intestinal Epithelial Cells In Vitro.

Zinc oxide nanoparticles (nano-ZnO) have diverse applications in numerous biomedical processes. The present study explored the effects of these nanoparticles on antioxidation, inflammation, tight junction integrity, and apoptosis in heat-stressed bovine intestinal epithelial cells (BIECs). Primary BIECs that were isolated and cultured from calves either were subjected to heat stress alone (42°C for 6 h) or were simultaneously heat-stressed and treated with nano-ZnO (0.8 μg/mL). Cell viability, apoptosis, and expression of genes involved in antioxidation (Nrf2, HO-1, SOD1, and GCLM), inflammation-related genes (TLR4, NF-κB, TNF-α, IL-6, IL-8, and IL-10), intestinal barrier genes (Claudin, Occludin, and ZO-1), and apoptosis-related genes (Cyt-c, Caspase-3, and Caspase-9) were assessed to evaluate the effect of nano-ZnO on heat-stressed BIECs. The nanoparticles significantly increased cell viability and decreased the rate of apoptosis of BIECs induced by heat stress. In addition, nano-ZnO promoted the expression of antioxidant-related genes HO-1 and GCLM and anti-inflammatory cytokine gene IL-10, and inhibited the pro-inflammatory cytokine-related genes IL-6 and IL-8. The nanoparticles also enhanced expression of the Claudin and ZO-1 genes, and decreased expression of the apoptosis-related genes Cyt-c and Caspase-3. These results reveal that nano-ZnO improve the antioxidant and immune capacity of BIECs and mitigate apoptosis of intestinal epithelial cells induced by heat stress. Thus, nano-ZnO have potential for detrimental the adverse effects of heat stress in dairy cows.

Tea polyphenols protect bovine intestinal epithelial cells from the adverse effects of heat-stress in vitro

Heat-stress (HS) leads to impaired gut health, adversely affecting milk production of dairy cows. In the present study, we investigated the protective effects of tea polyphenols (TP) against HS-induced damage in bovine intestinal epithelial cells (BIECs) and explored the underlying mechanisms. Primary BIECs were isolated from bovine duodenum, cultured and treated as follows: (1) control cells incubated in complete medium at 37 °C for 12 h, (2) TP group incubated in medium containing 100 μg/mL TP at 37 °C for 12 h, (3) HS group incubated in medium at 37 °C for 6 h followed by 6 h at 42 °C, and (4) HS + TP group incubated with 100 μg/mL TP for 6 h at 37 °C and 6 h at 42 °C. TP improved cell viability and antioxidant capacity, and decreased apoptosis and LDH activity. TP led to upregulation of Nrf2 and its target antioxidant genes HO-1, NQO1 and SOD1 expression. TP significantly decreased the expression of proinflammatory cytokine genes (NF-κB, IL-6 and TNF-α), and increased expression of the anti-inflammatory cytokine gene, IL-10. The above results suggested that TP protected BIECs from HS-induced adverse effects by alleviating oxidative stress and inflammatory responses, indicating that TP can alleviate HS-induced intestinal damage in dairy cows.

Expression of intra-tumoral necrosis-associated cytokine pattern correlated with prognosis and immune status in glioma.

Intra-tumoral necrosis (ITN) is reported to be an independent prognostic factor in glioma. However, knowledge of ITN is mainly limited to pseudopalisadwe, while its other aspects were neglected. Therefore, a deeper understanding of ITN could be valuable for understanding its exact role in glioma. The only reliable ITN model was time-dependently achieved with the GL261 syngeneic mouse model. The ITN-associated expression pattern was enriched from RNA sequencing. TCGA glioma samples were clustered into a high-expression group (HEG) and a low-expression group (LEG) based on their pattern and their association with prognosis, clinical status, immune status, and therapeutic responsiveness were compared. Mouse glioma with ITN demonstrated invasive histology. Cytokine signaling was significantly enriched in necrotic mouse glioma compared with non-necrotic glioma tissues. Nine pro-inflammatory (IL6, PPBP, IL1A, TNFSF11, CXCL11, CXCL9, CXCL10, CXCL3, and CCL8) and two anti-inflammatory cytokine (IL1RN and IL10) genes were found to be related to ITN-associated cytokine patterns. Comparative analysis showed that HEG had a significantly shorter survival time, five differentially distributed clinical statuses, more infiltrated immune cells, greater expression of immune checkpoints, and better therapeutic responsiveness than LEG. In conclusion, the ITN-associated cytokine pattern is characteristically expressed in glioma with ITN and might indicate necrosis missed in histology diagnosis. Its expression pattern could predict the prognosis, tumor grade, immune status, and therapeutic responsiveness of glioma patients.

Indoleamine 2, 3-dioxygenase 1 mediated alterations in the functionality of immune cells, deciphers the pregnancy outcomes in crossbred dairy cows

Pregnancy establishment in bovines requires maternal immune cell modulation. Present study investigated possible role of immunosuppressive indolamine-2, 3-dioxygenase 1 (IDO1) enzyme in the alteration of neutrophil (NEUT) and peripheral blood mononuclear cells (PBMCs) functionality of crossbred cows. Blood was collected from non-pregnant (NP) and pregnant (P) cows, followed by isolation of NEUT and PBMCs. Plasma pro-inflammatory (IFNγ and TNFα) and anti-inflammatory cytokines (IL-4 and IL-10) were estimated by ELISA and analysis of IDO1 gene in NEUT and PBMCs by RT-qPCR. Neutrophil functionality was assessed by chemotaxis, measuring activity of myeloperoxidase and β-D glucuronidase enzyme and evaluating nitric oxide production. Changes in PBMCs functionality was determined by transcriptional expression of pro-inflammatory (IFNγ, TNFα) and anti-inflammatory cytokine (IL-4, IL-10, TGFβ1) genes. Significantly elevated (P < 0.05) anti-inflammatory cytokines, increased IDO1 expression, reduced NEUT velocity, MPO activity and NO production observed only in P cows. Significantly higher (P < 0.05) expression of anti-inflammatory cytokines and TNFα genes were observed in PBMCs. Study highlights possible role of IDO1 in modulating the immune cell and cytokine activity during early pregnancy and may be targeted as early pregnancy biomarkers.

The effects of dietary supplementation with mushroom or selenium enriched mushroom powders on the growth performance and intestinal health of post-weaned pigs

BackgroundThere is an urgent need to identify natural bioactive compounds that can enhance gastrointestinal health and promote pig growth performance in the absence of pharmacological levels of zinc oxide (ZnO). The objectives of this study were to: 1) compare the effects of mushroom powder supplemented with inorganic selenium (inSeMP) to mushroom powder enriched with organic selenium (orgSeMP) to pharmacological levels of ZnO on growth performance and faecal scores (FS) for the first 21 d post-weaning (Period 1); and 2) compare the molecular and microbial effects of inSeMP and orgSeMP in these pigs on d 39 post-weaning (Period 2).MethodsIn Period 1, pigs (3 pigs/pen; 8 pens/treatment) were assigned to: (1) basal diet (control); (2) basal diet + zinc oxide (ZnO) (3100 mg/kg d 1–14, 1550 mg/kg d 15–21); (3) basal diet + mushroom powder supplemented with inorganic selenium (inSeMP) containing selenium (selenite) content of 0.3 mg/kg feed; (4) basal diet + mushroom powder enriched with organic selenium (orgSeMP) containing selenium (selenocysteine) content of 0.3 mg/kg feed. Mushroom powders were included at 6.5 g/kg of feed.ResultsIn Period 1, there was no effect of diets on average daily gain (ADG) and gain:feed (G:F) ratio (P > 0.05). The orgSeMP supplemented pigs had a lower average daily feed intake (ADFI) compared to all other groups (P < 0.05). The ZnO supplemented pigs had reduced FS compared to the basal and mushroom group, while the orgSeMP supplemented pigs had lower FS compared to the basal group during the 21 d experimental period (P < 0.05). In Period 2, there was no effect of diets on ADFI, ADG and G:F ratio (P > 0.05). The orgSeMP supplementation increased the caecal abundance of bacterial members of the Firmicutes and Bacteroidetes phylum, including Lactobacillus, Agathobacter, Roseburia, and Prevotella and decreased the abundance of Sporobacter compared to the basal group, while inSeMP increased the caecal abundance of Prevotella and decreased the caecal abundance of Sporobacter compared to the basal group (P < 0.05). Dietary supplementation with inSeMP increased expression of TLR4 and anti-inflammatory cytokine gene IL10 and decreased nutrient transporter gene FABP2 compared to the orgSeMP group (P < 0.05).ConclusionOrgSeMP is a novel and sustainable way to incorporate selenium and β-glucans into the diet of weaned pigs whilst improving FS and modulating the caecal microbiota.

УРОВЕНЬ ЭКСПРЕССИИ ГЕНОВ ПРО- И ПРОТИВОВОСПАЛИТЕЛЬНЫХ ЦИТОКИНОВ В КРОВИ КОРОВ ПРИ ГИПОФУНКЦИИ ЯИЧНИКОВ И ПРИМЕНЕНИИ ПРЕПАРАТА НА ОСНОВЕ ГМ-КСФ

The article presents the material for studying the expression level of pro- and anti-inflammatory cytokine genes in the blood of cows with ovarian hypofunction and the use of the drug based on gm-csf (granulocyte-macrophage colony-stimulating factor). It has been established that disorders in the functional activity of the genital gonads in cows occur against the background of an increase in the relative expression level of genes of pro-inflammatory cytokines (IL-1β - by 1.56 and 2.11 times, IL-2 - by 1.41 and 2.29 times, TNFα - by 19.5 and 48.8%), while reducing anti-inflammatory ones (IL-4 - by 27.6 and 37.9%, IL-10 - by 8.3 and 36.1%). An imbalance between the expression level of pro- and anti-inflammatory cytokines in the blood of cows is one of the etiological factors leading to hormonal imbalance and the development of hypofunctional ovarian disorders. The administration of a drug based on gm-csf to cows during insemination once at a dose of 5.0 ml to normalize the functional activity of the ovaries in case of their hypofunctional state with anovulation phenomena contributed to a decrease in the expression level of IL-1β by 27.3%, IL-2 – by 16.7%, TNFα – by 8.2%, with an increase in the expression of IL-4 by 4.8%, IL-10 – by 6.1%, which ensured a clinical efficacy of 86.7%. The use of a drug based on gm-csf in the cows with ovarian hypofunction, manifested by anaphrodisia, twice with an interval of 7 days at a dose of 10.0 ml provided a decrease in the expression level of IL-1β - by 1.9 times, IL-2 – by 38.5%, TNFα – by 14.8%, with an increase in IL-4 - by 11.1%, IL-10 – by 39.1% and a clinical efficacy of 90.0%.

МОЛЕКУЛЯРНО-КЛЕТОЧНЫЕ МАРКЕРЫ НЕЙРОВОСПАЛЕНИЯ И СОСТОЯНИЕ КОГНИТИВНЫХ ФУНКЦИЙ В ОТДАЛЕННЫЙ ПЕРИОД ПОСЛЕ ПРОЛОНГИРОВАННОГО γ-, n-ОБЛУЧЕНИЯ МЫШЕЙ В НИЗКИХ ДОЗАХ

The study aimed to investigate the effect of low-dose rate (2.13 mGy/h) prolonged γ,n-irradiation at 0.05 and 0.5 Gy on brain microglia, expression of pro- and anti-inflammatory cytokine genes, behavior and cognition of C57Bl/6 mice two months and one year after exposure. The decrease in the number of resting microglia and the increase in the proportion of activated microglia in brain cell preparations were observed after γ,n-irradiation at a dose of 0.5 Gy. Immunohistochemical analysis revealed the increased number of microglia cells and unaltered number of astrocytes in the hippocampus after γ,n-irradiation at both doses. Expression of TNFα and IL-1β pro-inflammatory cytokine genes in the hippocampus was increased after irradiation at both doses. Expression of TGFβ and IL-4 anti-inflammatory cytokine genes was decreased two months after irradiation only at a dose of 0.5 Gy, suggesting more intensive neuroinflammation. Analysis performed a year later revealed an age-related decrease in the number of resting microglia and increase in proportion of activated microglia compared to young animals, with more profound changes in male mice. The state of microglia in control and irradiated mice was similar one year after exposure. Neither two months nor one year after irradiation the impairments of motor activity and spatial memory of mice were detected; nevertheless, mice irradiated at a dose of 0.5 Gy demonstrated anxious behavior and decreased spatial learning in Morris water maze one year after exposure. In summary, we showed that γ, n-irradiation at doses of 0.05 and 0.5 Gy induced neuroinflammation two months after exposure. No difference from the control mice was observed one year after irradiation. Disturbances of behavior and spatial memory of irradiated mice were not detected.

Effects of cysteine addition to low-fishmeal diets on the growth, anti-oxidative stress, intestine immunity, and Streptococcus agalactiae resistance in juvenile golden pompano (Trachinotus ovatus).

As the precursor of taurine, cysteine serves physiological functions, such as anti-oxidative stress and immune improvement. Investigation of cysteine and its derivatives has made positive progress in avian and mammalian species, yet the study and application of cysteine in aquatic animals are relatively rare. Therefore, we evaluated the effects of supplementing a low-fishmeal diet with various levels of cysteine on the growth, antioxidant capacity, intestine immunity, and resistance against Streptococcus agalactiae of the juvenile golden pompano (Trachinotus ovatus). According to our study, exogenous supplementation with 0.6-1.2% cysteine greatly increased the final body weight (FBW) and specific growth rate (SGR) of golden pompano compared to the control group. Under the present conditions, the optimum dietary cysteine supplementation level for golden pompano was 0.91% based on the polynomial regression analysis of SGR. Meanwhile, we found that the Nrf2/Keap1/HO-1 signaling pathway was notably upregulated with the increase of exogenous cysteine, which increased antioxidant enzyme activity in serum and gene expression in the intestine and reduced the level of reactive oxygen species (ROS) in the serum of golden pompano. In addition, morphological analysis of the midgut demonstrated that exogenous cysteine improved muscle thickness and villi length, which suggested that the physical barrier of the intestine was greatly strengthened by cysteine. Moreover, cysteine increased the diversity and relative abundance of the intestinal flora of golden pompano. Cysteine suppressed intestinal NF-κB/IKK/IκB signaling and pro-inflammatory cytokine mRNA levels. Conversely, intestinal anti-inflammatory cytokine gene expression and serum immune parameters were upregulated with the supplementary volume of cysteine and improved intestine immunity. Further, exogenous cysteine supplementation greatly reduced the mortality rate of golden pompano challenged with S. agalactiae. In general, our findings provide more valuable information and new insights into the rational use of cysteine in the culture of healthy aquatic animals.

Comparative expression profiling of cytokine genes in Theileria annulata-infected and healthy cattle.

Theileriosis is one of the top ten economically important diseases in cattle in India. Cytokines are considered important mediators and regulators of the immune response to an infection. In the present study, the gene expression profiles of fourteen cytokines (IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL12A, IL12B, IL16, TGFB1, TNFA, IFNA and IFNB) were compared in Theileria annulata-infected and healthy crossbred cattle. Blood samples were obtained from the District Disease Diagnostic Laboratory, Karnal. The presence/absence of T. annulata infection in the animals was determined on the basis of blood smear examination and molecular detection through PCR using the genus-specific primers. Total RNA was extracted from peripheral blood mononuclear cells, which was further reverse transcribed to cDNA. Primer3 software was employed to design the primers for Real-Time qPCR. The results were examined using 2-∆∆Ctmethod with RPS15 and GAPDH as the reference genes. The expression of IL1B, IL6, IL8, IL10, IL12A, IL12B, TNFA, IFNA and IFNB was significantly higher, whereas the expression of IL2 was lower in the infected animals. The transcript levels of IL1A and TGFB1 were also higher in the diseased animals, but the results were non-significant. This study profiles the expression kinetics of various pro-inflammatory and anti-inflammatory cytokine genes in response to bovine theileriosis.

Cytokines in Lung Transplantation.

Lung transplantation has developed significantly in recent years, but post-transplant care and patients' survival still need to be improved. Moreover, organ shortage urges novel modalities to improve the quality of unsuitable lungs. Cytokines, the chemical mediators of the immune system, might be used for diagnostic and therapeutic purposes in lung transplantation. Cytokine monitoring pre- and post-transplant could be applied to the prevention and early diagnosis of injurious inflammatory events including primary graft dysfunction, acute cellular rejection, bronchiolitis obliterans syndrome, restrictive allograft syndrome, and infections. In addition, preoperative cytokine removal, specific inhibition of proinflammatory cytokines, and enhancement of anti-inflammatory cytokines gene expression could be considered therapeutic options to improve lung allograft survival. Therefore, it is essential to describe the cytokines alteration during inflammatory events to gain a better insight into their role in developing the abovementioned complications. Herein, cytokine fluctuations in lung tissue, bronchoalveolar fluid, peripheral blood, and exhaled breath condensate in different phases of lung transplantation have been reviewed; besides, cytokine gene polymorphisms with clinical significance have been summarized.

Effects of exogenous taurine supplementation on the growth, antioxidant capacity, intestine immunity, and resistance against Streptococcus agalactiae in juvenile golden pompano (Trachinotus ovatus) fed with a low-fishmeal diet.

Taurine has various biological functions in fish, playing an essential role in growth, resistance to oxidative stress, and intestine immunity. Here, we evaluated the effects of exogenous taurine added to low-fishmeal diets on the growth, anti-oxidative stress, intestine immunity, and Streptococcus agalactiae resistance in juvenile golden pompano (Trachinotus ovatus). Our study showed that exogenous taurine supplementation of 1.2% (T3 group) greatly enhanced the weight gain rate and specific growth rate (SGR) of juvenile golden pompano, significantly upregulating growth-related factor expression in the brain and liver, as well as the levels of growth-related parameters in the serum. Polynomial regression analysis using SGR estimated the optimal dietary taurine level for golden pompano at 1.18%. Moderate exogenous taurine also increased the muscular thickness and villus length within the intestine, maintained intestinal physical barrier stability, activated the Nrf2/Keap-1/HO-1 signaling pathway, increased intestinal antioxidant enzyme gene expression and antioxidant enzyme activity in the serum, and upregulated immunoglobulin and complement levels in parallel with declining reactive oxygen species (ROS) levels in the serum. Antioxidant factor expression was also upregulated in the intestine. Furthermore, supplementation suppressed NF-κB signaling and intestinal pro-inflammatory cytokine gene expression, increased anti-inflammatory cytokine gene expression, and improved intestine immunity. Finally, taurine supplementation improved the survival rate of golden pompano challenged with S. agalactiae. Overall, our findings provide additional information and support for the rational use of taurine in healthy aquatic animal farming.

Contrasting effects of sleep fragmentation and angiotensin-II treatment upon pro-inflammatory responses of mice

Disordered sleep promotes inflammation in brain and peripheral tissues, but the mechanisms that regulate these responses are poorly understood. One hypothesis is that activation of the sympathetic nervous system (SNS) from sleep loss elevates blood pressure to promote vascular sheer stress leading to inflammation. As catecholamines produced from SNS activation can directly regulate inflammation, we pharmacologically altered blood pressure using an alternative approach-manipulation of the renin-angiotensin system (RAS). Male C57BL6/J mice were treated with angiotensin or captopril to elevate and reduce blood pressure, respectively and then exposed to 24-h of sleep fragmentation (SF) or allowed to sleep (control). Pro- and anti-inflammatory cytokine gene expression and as endothelial adhesion gene expression as well as serum glucocorticoids (corticosterone) were measured. RAS manipulation elevated cytokines and endothelial adhesion expression in heart and aorta while SF increased cytokine expression in peripheral tissues, but not brain. However, there were interactive effects of angiotensin-II and SF upon cytokine gene expression in hippocampus and hypothalamus, but not prefrontal cortex. SF, but not RAS manipulation, elevated serum corticosterone concentration. These findings highlight the contrasting effects of RAS manipulation and SF, implying that inflammation from SF is acting on different pathways that are largely independent of RAS manipulation.