Serum Concentrations Of IL-1β Research Articles

Improvement of weanling pigs immune status and metabolic condition using ultraweak light.

Weaning stress is the most common issue in swine farms, which increases mortality and morbidity. The use of artificial light is an option for modifying the immune system and metabolic pathways. This study aimed to evaluate the influence of ultraweak light (Photonia) on growth performance, immune system and metabolism of weanling pigs, and the carry-over effect on the growth performance in postweanling growing stages. A total of 30 weaned pigs with an average initial body weight of 7.06 ± 0.11 kg (age: 21 days) were allotted two treatments (Control and Photonia) with 15 replicates. The pelleted form diets were prepared for pigs in three phases including phase 1 (Days 0-14), phase 2 (Days 15-28) and phase 3 (Days 29-48). The gain-to-feed ratio (G:F) of pigs was significantly greater in the Photonia treatment. On Day 28, a higher concentration of immunoglobin A (IgA) (p < 0.01) and IgG (p < 0.01) was observed in the Photonia pigs. On Day 48, the Photonia treatment showed a greater serum IgA (p < 0.01) and IgG (p < 0.05). The concentration of interleukin (IL)-6 was decreased (p < 0.05) in the Photonia treatment. At Day 48, the concentrations of tumour necrotic factor-α, IL-1β and IL-6 in serum were decreased (p < 0.05) in pigs in the Photonia treatment. Metabolic pathways analysis showed that the Photonia treatment increased the d-glutamine, d-glutamate, alanine, aspartate, glutamate and phenylalanine compared with the control treatment. In conclusion, the use of Photonia for weanling pigs is recommended due to improved G:F, immune status and activation of amino acids metabolic pathways including d-glutamine, d-glutamate, alanine, aspartate, glutamate and phenylalanine.

β-mannanase supplemented in diets saved 85 to 100 kcal of metabolizable energy/kg, supporting growth performance and improving nutrient digestibility in grower pigs

The effects of β-mannanase supplementation in metabolizable energy (ME)-reduced diets containing xylanase-phytase were investigated on growth performance, fecal score, ultra-sounded backfat thickness and loin depth, blood profile, apparent total tract digestibility (ATTD), digesta passage rate, and fecal microbiome in grower pigs (n = 40, 26.09 ± 0.96 kg) randomly assigned within 4 treatments: a control diet containing isolated phytase and xylanase valued at 40 kcal of ME/kg (CD0), CD0 + β-mannanase (0.3 g/kg valued at 30 kcal of ME/kg) (CD70), CD0 + β-mannanase (0.3 g/kg valued at 45 kcal of ME/kg) (CD85), and CD0 + β-mannanase (0.3 g/kg valued at 60 kcal of ME/kg) (CD100). Growth performance was not affected in pigs fed ME-reduced diets containing β-mannanase. Pigs with CD100 had lower serum IL-1β concentration, and higher IL-10 was observed in pigs on CD0 than those fed β-mannanase. Coefficients of ATTD, and ATTD of DM and CP were higher in animals fed CD85 or CD100. Pigs with CD85 had higher alpha diversity richness but lower Firmicutes:Bacteroidota ratio. Acidaminococcaceae and Ruminococcaceae were more abundant in pigs fed CD0, but lower for Christensenellaceae NSJ-63 and NSJ-63 sp014384805. Pigs in CD85 showed higher Bacteroidaceae and Prevotella abundance, and lower for Streptococcaceae and Streptococcus. In conclusion, supplementation of β-mannanase in diets containing xylanase-phytase saved 85 to 100 kcal of ME/kg by supporting growth performance and improving nutrient digestibility in grower pigs.

Maternal Chronic Ultrasound Stress Provokes Immune Activation and Behavioral Deficits in the Offspring: A Mouse Model of Neurodevelopmental Pathology.

Neurodevelopmental disorders stemming from maternal immune activation can significantly affect a child's life. A major limitation in pre-clinical studies is the scarcity of valid animal models that accurately mimic these challenges. Among the available models, administration of lipopolysaccharide (LPS) to pregnant females is a widely used paradigm. Previous studies have reported that a model of 'emotional stress', involving chronic exposure of rodents to ultrasonic frequencies, induces neuroinflammation, aberrant neuroplasticity, and behavioral deficits. In this study, we explored whether this model is a suitable paradigm for maternal stress and promotes neurodevelopmental abnormalities in the offspring of stressed females. Pregnant dams were exposed to ultrasound stress for 21 days. A separate group was injected with LPS on embryonic days E11.5 and E12.5 to mimic prenatal infection. The behavior of the dams and their female offspring was assessed using the sucrose test, open field test, and elevated plus maze. Additionally, the three-chamber sociability test and Barnes maze were used in the offspring groups. ELISA and qPCR were used to examine pro-inflammatory changes in the blood and hippocampus of adult females. Ultrasound-exposed adult females developed a depressive-like syndrome, hippocampal overexpression of GSK-3β, IL-1β, and IL-6 and increased serum concentrations of IL-1β, IL-6, IL-17, RANTES, and TNFα. The female offspring also displayed depressive-like behavior, as well as cognitive deficits. These abnormalities were comparable to the behavioral changes induced by LPS. The ultrasound stress model can be a promising animal paradigm of neurodevelopmental pathology associated with prenatal 'emotional stress'.

Issues of cytokine regulation in vibration disease and associated conditions

The article presents the data on the state of cytokine regulation and indicators of endothelial damage when exposed to industrial vibration (general or local) and in combination with arterial hypertension. Changes in the levels of pro-inflammatory and anti-inflammatory cytokines are described, and biomarkers of endothelial dysfunction in the combined course of vibration disease and arterial hypertension are investigated. The study of the cytokine profile and biomarkers of damage to the vascular endothelium in this pathology will allow making early diagnosis of vascular disorders and optimizing preventive measures for workers in vibration hazard industries. Purpose of the study. Improving the quality of early diagnosis and prevention of vibration dis ease in an isolated course and its combination with arterial hypertension based on a study of the cytokine profile and biomarkers of endothelial dysfunction in this pathology. Material and research methods. A comprehensive study included 84 patients with vibration disease from the exposure to local and general vibration of the first and second degree, 61 patients with a combined course of vibration disease from the exposure to local and general second degree vibration and arterial hypertension, and 30 participants in the control group not exposed to industrial vibration and healthy according to medical examination. The levels of pro-inflammatory (IL-1β, IL-8, TNF-α) and anti-inflammatory cytokines (IL-4) and the biomark ers of endothelial damage (EDN1, TGF-β1, VEGF-A, PDGF-BB, fibronectin, and von Willebrand factor) were determined by enzyme immunoassay. Results. The response of the immune system to the effects of industrial vibration is characterized by a cytokine imbalance — an increase in the level of pro-inflammatory cytokines (IL-1β, IL-8, TNFα) and a decrease in the level of anti-inflammatory cytokine (IL-4). With a combined course of vibration disease and arterial hypertension, the cytokine imbalance is characterized by an even more significant increase in serum concentration of IL-1β, IL-8, TNFα and a decrease in serum IL-4 concentration. Endothelial dysfunction in vibration disease from the exposure to both local and general vibration in combination with arterial hypertension is characterized by a significant increase in the serum concentration of EDN1, TGF-β1, VEGF-A, PDGF-BB, fibronectin, and von Willebrand factor. Conclusion. The study of the cytokine profile and biomarkers of damage to the vascular endothelium in this pathology will allow making early diagnosis of vascular disorders and optimizing preventive measures for workers in vibration hazard industries.

#5599 FEBUXOSTAT IN COMBINATION WITH ANAKINRA OR PH NEUTRALIZATION IMPROVE THE OUTCOMES IN HYPERURICEMIA-RELATED URIC ACID CRYSTALLURIA IN MICE

Abstract Background and Aims Currently, it is unknown whether CKD patients with hyperuricemia (HU)-related uric acid (UA) crystalluria, also known as chronic UA nephropathy, would benefit from targeting HU, UA crystallization, and/or inflammation to slow down the progressive decline of kidney function. Although large multi-center RCTs with xanthine oxidase inhibitors (XOIs) such as allopurinol and two Mendelian randomization studies have disproven a causal link between HU and CKD progression, one cannot rule out that XOIs may still represent a viable therapeutic approach for patients with HU-related UA crystalluria in order to prevent CKD progression. This issue requires clarification. To address this, we used our well-established CKD mouse model of HU with UA crystalluria and tested numerous therapeutic approaches including XOI and anakinra. Method Alb-creERT2;Glut9lox/lox (male and female) mice were injected with tamoxifen and placed on an acidogenic diet with the purine inosine to induce HU and crystalluria-related CKD. After chronic UA nephropathy was established on day 14, mice were treated either 1) with the XOIs allopurinol or febuxostat, 2) with sodium bicarbonate supplementation to neutralize urinary pH to prevent UA crystallization, or 3) with anakinra to inhibit IL-1β-driven inflammation, or 4) a combination of the aforementioned therapies for 2 weeks. Saline was used as control. On day 0, 14, and 28, GFR (as primary endpoint) as well as serum UA levels and urinary UA crystals (as secondary endpoints) were determined. At the end of the study, we quantified kidney injury, immune cell infiltration, inflammation, and interstitial fibrosis using histological analysis, ELISA, RT-qPCR, and flow cytometry. Results Therapy with XOIs but not with sodium bicarbonate and anakinra reduced serum UA levels in mice with HU and UA crystalluria. Treating mice with febuxostat improved kidney function (increase in GFR) by preventing kidney injury, UA crystalluria, UA crystal granuloma formation, and interstitial fibrosis compared with the control group, while allopurinol worsened the outcomes of chronic UA nephropathy. On the other hand, sodium bicarbonate supplementation had no effects on serum UA levels, kidney inflammation, immune cell infiltration and interstitial fibrosis but improved kidney function due to less urinary UA crystal deposition (neutralization of urine pH) and granuloma formation as compared with saline-treated mice. As expected, anakinra did not slow down the progression of chronic UA nephropathy as noticed by a similar GFR decline compared with the control group. Although, anakinra reduced the number of infiltrating immune cells and the mRNA expression of inflammatory mediators in the kidney as well as the serum IL-1β concentrations, we observed more interstitial fibrosis. In combination, allopurinol with anakinra did not improve the outcomes of chronic UA nephropathy despite the inhibitory effect of anakinra on the inflammatory response because allopurinol triggered tubulointerstitial nephritis and vasodilation. Interestingly, both combination therapies febuxostat with anakinra and febuxostat with sodium bicarbonate were protective by improving kidney function and preventing UA crystal deposition, kidney injury, inflammation, interstitial fibrosis and UA crystal granuloma formation compared with the single therapies. Conclusion Our interventional study is the first pre-clinical study, which reveals that the XOIs allopurinol and febuxostat have differential effects on the outcomes of HU-related UA crystalluria. While allopurinol contributes to the progression of CKD by causing tubulointerstitial nephritis in mice, febuxostat rather seems to be renoprotective similar to urinary pH neutralization with sodium bicarbonate. We identified both combination therapies febuxostat with anakinra or sodium bicarbonate as most beneficial in order to prevent CKD progression in mice. Thus, febuxostat in combination with IL-1β inhibition or prevention of urinary UA crystallization may represent therapeutic approaches for patients with chronic UA nephropathy.

Inflammaging in domestic dogs: basal level concentrations of IL-6, IL-1β, and TNF-α in serum of healthy dogs of different body sizes and ages.

Inflammaging, a "hallmark of aging," refers to a chronic, progressive increase in the proinflammatory status of mammals as they age, and this phenotype has been associated with many age-related diseases, including cardiovascular disease, arthritis and cancer. Though, inflammaging research is common in humans, there is a lack of data for this process for the domestic dog. Here, serum concentrations of IL-6, IL-1β and TNF-α in healthy dogs of different body sizes and ages were measured to determine whether inflammaging may play a mechanistic role in aging rates in dogs, similar to those found in humans. Using a four-way ANOVA, a significant decrease in IL-6 concentrations in young dogs with the rest of the age categories showing increased IL-6 concentrations was found, similar to humans. However, only young dogs have decreased IL-6 concentrations, with adult dogs having similar IL-6 concentrations to senior and geriatric dogs, implying differences in aging rates between humans and dogs. And, there was a marginally significant interaction between sex*spayed or neutered status and IL-1β concentrations with intact females having the lowest IL-1β concentrations compared with intact males, and spayed and neutered dogs. The presence of estrogen in intact females may, overall, decrease inflammatory pathways. This implies that age at spaying or neutering may be an important aspect to consider for inflammaging pathways in dogs. Furthermore, sterilized dogs often die of immune-related diseases, which could be linked to the increases in IL-1β in sterilized dogs found in this study.

Soluble interleukin-2 receptor combined with interleukin-8 is a powerful predictor of future adverse cardiovascular events in patients with acute myocardial infarction.

Little is known about the role of interleukin (IL) in patients with acute myocardial infarction (MI), especially soluble IL-2 receptor (sIL-2R) and IL-8. We aim to evaluate, in MI patients, the predictive value of serum sIL-2R and IL-8 for future major adverse cardiovascular events (MACEs), and compare them with current biomarkers reflecting myocardial inflammation and injury. This was a prospective, single-center cohort study. We measured serum concentrations of IL-1β, sIL-2R, IL-6, IL-8 and IL-10. Levels of current biomarkers for predicting MACEs were measured, including high-sensitivity C reactive protein, cardiac troponin T and N-terminal pro-brain natriuretic peptide. Clinical events were collected during 1-year and a median of 2.2 years (long-term) follow-up. Twenty-four patients (13.8%, 24/173) experienced MACEs during 1-year follow-up and 40 patients (23.1%, 40/173) during long-term follow-up. Of the five interleukins studied, only sIL-2R and IL-8 were independently associated with endpoints during 1-year or long-term follow-up. Patients with high sIL-2R or IL-8 levels (higher than the cutoff value) had a significantly higher risk of MACEs during 1-year (sIL-2R: HR 7.7, 3.3-18.0, p < 0.001; IL-8: HR 4.8, 2.1-10.7, p < 0.001) and long-term (sIL-2R: HR 7.7, 3.3-18.0, p < 0.001; IL-8: HR 4.8, 2.1-10.7, p < 0.001) follow-up. Receiver operator characteristic curve analysis regarding predictive accuracy for MACEs during 1-year follow-up showed that the area under the curve for sIL-2R, IL-8, sIL-2R combined with IL-8 was 0.66 (0.54-0.79, p = 0.011), 0.69 (0.56-0.82, p < 0.001) and 0.720 (0.59-0.85, p < 0.001), whose predictive value were superior to that of current biomarkers. The addition of sIL-2R combined with IL-8 to the existing prediction model resulted in a significant improvement in predictive power (p = 0.029), prompting a 20.8% increase in the proportion of correct classifications. High serum sIL-2R combined with IL-8 levels was significantly associated with MACEs during follow-up in patients with MI, suggesting that sIL-2R combined with IL-8 may be a helpful biomarker for identifying the increased risk of new cardiovascular events. IL-2 and IL-8 would be promising therapeutic targets for anti-inflammatory therapy.

Low vitamin D status is associated with inflammatory response in older patients with cerebral small vessel disease.

This study aimed to determine the association of the NF-κB inflammatory signaling pathway with vitamin D status in older cerebral small vessel disease (SVD) patients. We measured serum 25(OH)D, pro-and anti-inflammatory cytokines, and mRNA levels of the vitamin D-activating enzyme, CYP27B1, as well as NF-kB, COX-2, the chemokine-CCL2, IL-1β, IL-6, TNF-α, TGF-β, and IL-10, in cerebral SVD patients aged ≥60years presenting with vascular dementia and age and gender-matched healthy controls. Low vitamin D status (insufficiency: serum 25(OH)D 12-20ng/ml; deficiency: ≤12ng/ml) was more prevalent among patients compared to controls. The mRNA levels of NF-kB, COX-2, CCL2, IL-1β, and IL-6, and serum levels of pro-inflammatory cytokines (IL-1α, IL-1β, IL-6, and TNF-α) were significantly higher in cases compared to controls. There was a significant correlation between CYP27B1 and NF-kB, COX-2, CCL2, and IL-1β gene expression. Serum IL-1α, IL-1β, and IL-6 concentrations and the expression of CCL-2, NF-kB2, and NF-kB3 genes were higher in vitamin D-deficient subjects compared to vitamin D-sufficient subjects. There was a significant negative correlation between serum 25(OH)D and IL-1α, IL-6, and TNF-α, and a positive correlation between 25(OH)D and IL-10. Low vitamin D is associated with an inflammatory response via NF-kB signaling, which could play a role in the etio-pathogenesis of SVD. Further large-scale studies are required to validate our findings.

Long term dietary vitamin D3 supplementation impacts both microbicidal and inflammatory responses to ex-vivo Mycobacterium bovis BCG challenge in dairy calves

Vitamin D deficiency (VDD) is associated with enhanced susceptibility to multiple respiratory diseases in humans, including tuberculosis. However, the consequences of VDD for disease susceptibility in calves are unknown. Previously we developed a model to drive divergent circulating 25OHD concentrations in cattle, where animals were supplemented with vitamin D3 (vit D3) from birth to 7 months of age. Calves in the control group (Ctl) received a diet containing a standard vit D3 concentration, whereas the vit D group (VitD) received a diet with the highest vit D3 concentration allowed under EU guidelines. Here, we assessed the microbicidal activity and immunoregulatory effect of divergent 25OHD circulating levels to Mycobacterium bovis BCG challenge ex-vivo. Blood samples from Ctl and VitD calves were taken at 1-, 3- and 7-months of age. 25OHD concentrations were significantly different at 7 months (but not at 1 or 3 months) with animals from the VitD group having higher serum levels. Differences in microbicidal activity followed the same pattern, with no significant differences observed at 1 and 3 months, but at 7 months a significant increase in the percentage of bacteria killed was detected. Furthermore, analysis of the reactive oxygen species (ROS) and nitric oxide (NO) in serum showed a higher production of ROS and NO in VitD-supplemented calves. In contrast, serum concentrations of IL-1β and IL-8 were significantly lower. A similar anti-inflammatory profile was observed after gene expression analysis, with a significant downregulation of a cluster of genes including IL1B, IL1R1, CXCL1, CXCL2, CXCL5, MMP9 and COX2 and an upregulation of CXCR1, CX3CR1 and NCF1, in VitD calves after BCG challenge relative to Ctl animals. Collectively, these results suggest that dietary vit D3 boosts antimicrobial and innate immune responses and thereby could improve host anti-mycobacterial immunity.

Определение цитокинов в сыворотке крови и во влаге передней камеры у пациентов с глаукомой и катарактой на фоне псевдоэксфолиативного синдрома

Purpose. To investigate changes in proinflammatory cytokines Interleukin (IL 1β) and Tumor necrosis factor (TNF-α) before surgery in anterior chamber moisture and in blood serum in cataract and in its combination with glaucoma due to pseudoexfoliative syndrome (PES). Material and methods. The results of the examination included 61 patients aged 56 to 86 years (average age 72.2 years) who were hospitalized for cataract surgery. Among them, 12 (19.7 %) people had primary open-angle pseudoexfoliative glaucoma with normalized intraocular pressure (IOP). A group of 34 patients were diagnosed with PES, but without the presence of glaucoma. The control group consisted of 15 patients with cataracts of comparable age without signs of PES. The analysis of the results took into account the data of ophthalmological examination, immunological studies, namely measurements of the concentration of IL-1ß and TNFa in blood serum and in the moisture of the anterior chamber, as well as somatic status. Results. PES, in general (i.e. in the study of 46 patients), occurred in 23.9 % of cases, stage II — in 43.5 % and stage III — in 32.6%. With a combination of cataracts and glaucoma (in the advanced stage), PES stage I was diagnosed more often — in 33.3 %.Inflammatory postoperative complications were diagnosed only in stages II or III of PES, and more often with a combination of glaucoma and cataract (in 33.3 % of cases) than with isolated surgical treatment of cataract (11.8 %). There was a statistically significant (p &lt; 0.05) increase in the indicators of proinflammatory cytokines IL- 1ß and TNF-α in blood serum and especially in the moisture of the anterior chamber in patients with PES in comparison with the age control group. Various concomitant somatic pathologies in patients with PES, especially in the presence of glaucoma, occurred more often (93.5 %) than in patients without pseudoexfoliation (73.3 %). Conclusion. Significantly increased, in comparison with the control rate, concentrations of proinflammatory cytokines IL-1β and TNFa in blood serum and moisture of the anterior chamber of the eyes with unoperated cataract, and especially when combined with glaucoma due to PES, correlated with the degree of its development and the risk of postoperative, including inflammatory, complications. Taking into account more frequent presence of somatic pathology in patients with PES, it is necessary to have more frequent dynamic follow-up after surgery of patients with PES for the purpose of early diagnosis and timely treatment of potential complications. Keywords: cataract, glaucoma, surgical treatment, pseudoexfoliative syndrome, proinflammatory cytokines Interleukin (IL-1β) and Tumor necrosis factor (TNF-α), postoperative, inflammatory complications

Pueraria lobata polysaccharides alleviate neonatal calf diarrhea by modulating gut microbiota and metabolites.

Neonatal calf diarrhea (NCD) is still one of the most critical diseases in calf rearing. Studies have shown that Pueraria lobata polysaccharides (PLP) have intense antioxidant and immunomodulatory activity and modulate gut microbiota. This randomized clinical trial aimed to determine the effect of PLP on the neonatal calf with diarrhea. In this study, we recorded the fecal score of experimental calves, and calves with fecal scores ≥ 2 were determined as diarrhea and assessed their serum concentrations of inflammatory cytokines and oxidative damage-related indices. Fecal microbiota and metabolomics of diarrheal calves were further investigated. Results showed that treatment with PLP decreased the fecal score of diarrheal calves, serum concentrations of IL-1β, TNF-γ, and malondialdehyde, and also elevated the level of superoxide dismutase. In addition, PLP treatment altered the gut microbiota, significantly increased the relative abundances of beneficial bacteria, including the phyla Bacteroidetes and Actinobacteria, the genus Collinsella, Megamonas, and Bifidobacterium; decreased the relative abundances of pathogenetic or diarrhea related bacteria, such as Proteobacteria, Fusobacteria, Clostridium_sensu_stricto_1, and Escherichia_Shigella. Moreover, PLP can increase the fecal concentrations of isobutyric acid, propionic acid, and pantothenate; lower the levels of PC [18:0/18:1(9Z)], arachidonic acid, and docosahexaenoic acid. Thus, the results suggested that the PLP may perform the therapeutic activity via alleviating intestinal inflammation and regulating gut microbiota, avoiding further dysbiosis to restore the metabolism of gut microbiota, and finally promoting the recovery of diarrhea. The change further mitigated intestinal inflammation and oxidative damage in diarrheal calves. This indicated that PLP might be a promising treatment to attenuate diarrhea in neonatal calves.

Enhancement of oral bioavailability and anti-colitis effect of luteolin-loaded polymer micelles with RA (rosmarinic acid)-SS-mPEG as carrier

Objective Polymer micelles were prepared (L-RSPMs) with luteolin and synthetic RA-SS-mPEG polymeric material before evaluation of their anti-inflammatory effect on 2, 4, 6-trinitro-benzene-sulfonic acid (TNBS)-induced ulcerative colitis (UC) model in rats. Methods The synthetic RA-SS-mPEG was characterized with NMR spectroscopy, before preparation of luteolin-coated RA-SS-mPEG polymer micelles. The in vitro characterization and evaluation of the formulation were accomplished, couple with its pharmacokinetic parameters. The levels of PEG2, MDA, CRP and GSH, as well as concentrations of TNF-α, IL1-β, IL-6 and IL-10 in serum and colon tissue were detected via ELISA kit. The degree of colon injury and inflammation was evaluated via histopathologic examination. Results L-RSPMs displayed small average droplet size (133.40 ± 4.52 nm), uniformly dispersed (PDI: 0.163 ± 0.011), good stability, slow release and enhanced solubility. We observed 353.28% increase in the relative bioavailability of L-RSPMs compared to free luteolin, while the half-life of the micelle was extended by 6.16h. Compared to model (M) group, luteolin (low and high doses) and L-RSPMs (low and high doses) significantly reduced levels of MDA, PEG2, CRP, TNF-α, IL-6 and IL-1β in colon tissue and serum of colitic rats but dose dependently increased IL-10 and SOD levels (p < 0.01). Histopathologic examination of colon showed that luteolin (low and high doses) and L-RSPMs (low and high doses) improved colonic inflammation in colitic rats to varying degrees compared to M group. Conclusion L-RSPMs could improve TNBS-induced colon inflammation by enhancing bioavailability, promoting antioxidant effects and regulating cytokine release, which may become a potential agent for UC treatment in clinical settings.

Inhibitory effects of fluoxetine on the secretion of inflammatory mediators and JAK/STAT3 and JNK/TLR4 gene expression.

Selective serotonin reuptake inhibitors (SSRIs) are the most common class of medicines used for the treatment of major depression. Recent studies have reported an association between depression and inflammation and suggested the significant effects of SSRIs on inflammatory processes. The current study aimed to evaluate the effects of fluoxetine, an SSRI, on the level of inflammatory cytokines, including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), in the rat serum and RAW264.7 mouse macrophage cell line, using ELISA sandwich assays. Also, the expression of inflammatory genes, including JAK/STAT3 and TLR4/JNK, was examined in macrophages, using real-time quantitative reverse transcription PCR to determine the potential mechanism of fluoxetine in inflammation. The rats received fluoxetine (10, 20, and 40mg/kg) 30min before lipopolysaccharide (LPS) treatment for 90min. The cells received different doses of fluoxetine (5, 10, and 20µg/mL) before stimulation with LPS for 24 or 48h. The serum concentrations of IL-1β, IL-6, and TNF-α were reduced in rats and cells treated with fluoxetine. Following fluoxetine administration, the expression of JAK/STAT3 and TLR4/JNK genes was significantly decreased in the RAW264.7 cells treated with LPS for 24h. However, after 48h of treatment with LPS, fluoxetine failed to diminish the elevated expression of JAK and JNK genes, while it significantly decreased the expression of STAT3 and TLR4 genes. The findings revealed that fluoxetine has anti-inflammatory properties, mainly due to the reduction of inflammatory cytokines and inhibition of JAK/STAT3 and TLR4/JNK gene expression in macrophages.

ПРОГНОСТИЧЕСКИЕ ПРЕДИКТОРЫ ТЯЖЕСТИ ТЕЧЕНИЯ КОРОНАВИРУСНОЙ ИНФЕКЦИИ У ДЕТЕЙ ДОШКОЛЬНОГО ВОЗРАСТА

As for adults, the prognostic predictive factors have been established, including some cytokines, associated with the diagnosis, pathogenesis, and severity of the coronavirus infection (COVID-19). However, predictors for the severity of the course of coronavirus infection in children are not well understood. In addition, there is no information on the cytokine profile in preschool children with COVID-19 as yet. The purpose of the research was to create a predictive model based on a comprehensive analysis of cytokine levels, which would then allow predicting the severity of the course of coronavirus infection in preschool children. Materials and methods used: 21 children (47.6% (10/21) boys and 52.4% (11/21) girls) aged 4.0 [3.0; 4.0] y/o with confirmed coronavirus infection (main group) vs. 25 children (44% (11/25) boys and 56% (14/25) girls) of health group I aged 3.0 [2.9; 3.7] y/o (the control group). Depending on the severity of COVID-19, patients of the main group were divided into subgroups: SG1 of 7 children with asymptomatic COVID-19; SG2 of 7 patients with a mild form; and SG3 of 7 cases of moderate form of the disease. In all children, the presence of SARS-CoV-2 in nasopharyngeal and oropharyngeal swabs was detected by PCR. The concentration of cytokines in blood serum was assessed by the flow fluorometry. Results: in children with COVID-19 at the time of admission to the hospital, the serum concentration of IL-1β, IL-2, IL-4, IL-6, IL-8, IP-10, MCP-1, IFN-γ, and TGF-β1 was statistically significantly higher than in the control group (p&lt;0.05). Thus, based on a comprehensive analysis of the levels of IL-1β, IL-2, IL-6, IL-8, IL-12p70, IP-10, and TGF-β1 at the time of admission to the hospital, the prognostic model was developed allowing predicting the severity of COVID-19 in preschool children (Se=1.0 [95% CI 0.99-1.00], Sp=0.98 [95% CI 0.96-0.99], ROC-AUC=1.0 [95% CI 0.99-1.00], p&lt;0.001; R2=0.93). Conclusion: the results of a comprehensive analysis allow us to consider the indicators of IL-1β, IL-2, IL-6, IL-8, IL-12p70, IP-10, and TGF-β1 as predictors for coronavirus infection severity in preschool children.

Treadmill training mitigates bone deterioration via inhibiting NLRP3/Caspase1/IL-1β signaling in aged rats

IntroductionAlthough aerobic physical exercise may improve osteoporosis during ageing, the underlying mechanism of the favorable effects remains unclear. The aim of this study was to examine the localized and generalized proinflammatory indicators and the adaptive skeletal responses to treadmill training in aged rats to explore the potential mechanisms by which treadmill training impacts bone deterioration in a natural aged rat model.Materials and methodsA total of 24 Sprague Dawley (SD) rats were included in this study. Sixteen of all these animals were twenty-four months natural aged male SD rats, which were distributed into two groups (n = 8/group): AC group with sham treadmill training, and AT group with 8 weeks treadmill training. The remaining 8 were six months male SD rats matched subline and supplier, which were used as the adult control group with sham treadmill training (YC group, n = 8). The serum, bone marrow, fresh femur, tibia, and lumbar spine were harvested for molecular biological analysis, bone mineral density (BMD) testing, and micro-CT analysis after 8 weeks of treadmill training.ResultsAfter 8 weeks of intervention, the results showed that treadmill training increased BMD and inhibited deterioration of bone microarchitecture of hind limb bones. Further analysis showed that treadmill training increased serum P1CP concentration and decreased serum CTX-1level. Interestingly, treadmill training down-regulated the protein expressions of proinflammatory indicators, including NLRP3, proCaspase1, cleaved Caspase1, IL-1β, and GSDMD-N, and the mRNA levels of NLRP3, Caspase1, and IL-1β of the bone marrow. In addition, treadmill training also inhibited serum TNF-α and IL-1β concentration. However, 8 weeks of treadmill training did not increase BMD and bone microarchitecture in the lumbar spine.ConclusionTreadmill training mitigates the ageing-induced bone loss and reverses the deterioration of bone microarchitecture in hind limbs probably through inhibiting NLRP3/Caspase1/IL-1β signaling to attenuate low-grade inflammation and improve the inflammatory bone microenvironment.

Determining Serum Amyloid-A (SAA), Haptoglobin (Hp), Tumor Necrosis Factor (TNF-α) and Interleukins-1 (IL-1β) and 6 (IL-6) Levels in Neonatal Calves with Amoebiasis

Background: Amoebiasis is a pathogen organism which is well-known both in human and veterinary medicine. The aim of this study was to determine serum amyloid-A (SAA), haptoglobin (Hp), tumor necrosis factor (TNF-α) and interleukins-1 (IL-1β) and 6 (IL-6) levels in neonatal calves which were naturally infected with amoebiasis. Methods: In the study, 40 one to four-week-old calves with amoebiasis which were clinically diagnosed with diarrhea and were non-responsive to the treatment, and 10 healthy calves were used. From the stool samples taken from the calves which were clinically diagnosed with diarrhea, Entamoeba histolytica was detected by using ELISA kit. Serum was obtained from the blood taken from vena jugularis from all calves. Concentrations of SAA, Hp, TNF-α, IL-1β and IL-6 in serum were measured in ELISA by using commercial kits. Also, some enzymes measurements were made from serum. Result: Concentrations of SAA, Hp, TNF-α, IL-1β and IL-6 in calves with amoebiasis were higher (p less than 0.05) in comparison to the control group. To conclude, amoebiasis led to an increase in the concentrations of SAA, Hp, TNF-α, IL-1β and IL-6 in neonatal calves. We suggest that this might have stemmed from the inflammatory response against amoebiasis.

Dietary 25-hydroxyvitamin D improves productive performance and intestinal health of laying hens under Escherichia coli lipopolysaccharide challenge

The effect of 25-hydroxyvitamin D (25OHD) on the immune response of laying hens is not well elucidated. This study investigated the effects of 25OHD on egg production, egg quality, immune response, and intestinal health of laying hens challenged with Escherichia coli lipopolysaccharide (LPS). One hundred and sixty laying hens at 45 wk of age were randomly divided into 4 dietary treatments with 10 replicates of 4 birds. Hens were fed the corn-soybean based diets contained either 0 or 80 µg/kg 25OHD for 8 wks. At wk of 53 wk, birds of each dietary treatment were injected into the abdomen with 1.5 mg/kg body weight of either LPS or saline a day at 24-h intervals for continuous 7 d. LPS injection significantly decreased (PLPS < 0.05) egg laying rate, feed intake and feed efficiency; while the supplementation of 25OHD increased (PInteraction < 0.05) egg laying rate, feed efficiency and decreased (PInteraction < 0.05) the broken egg rate in layers under LPS injection. LPS challenge decreased (PLPS < 0.05) eggshell strength, eggshell thickness, albumen height and Haugh unit, while dietary 25OHD supplementation increased eggshell strength and eggshell thickness (P25OHD < 0.05). The serum proinflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6)], endotoxin and diamine oxidase (DAO) levels were higher in layers under LPS challenge (PLPS < 0.05); whereas the dietary addition of 25OHD were shown to decrease (P25OHD < 0.05) serum IL-1β and IL-6 concentration irrespective of LPS challenge and led to a higher serum 25OHD level and a reduction in endotoxin concentration in layers under LPS challenge (PInteraction < 0.05). The layers under LPS challenge had higher crypt depth and lower villus height/crypt depth (V/C) ratio in duodenum and jejunum (PLPS < 0.05), while feeding 25OHD were shown to have decreasing effect on crypt depth and increasing effect V/C ratio in layers under LPS challenge (PInteraction < 0.05). Layers under LPS challenge had lower mRNA expression of intestinal barrier associated proteins (claudin-1 and mucin-1) (PLPS < 0.05), while the addition of 25OHD up-regulated claudin-1 and mucin-1 expression (Pinteraction < 0.05). Lower antioxidant enzymes activities, including superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), glutathione peroxidase (GPx) and higher malondialdehyde (MDA) content in jejunum were found in layers challenged with LPS (P25OHD < 0.05). The effect of 25OHD reversed the effect of LPS on SOD, T-AOC, and MDA content (PInteraction< 0.05). These results suggest that supplementing 80 µg/kg 25OHD in diets may elevate laying performance and egg quality through the improvement of intestinal barrier function, antioxidant capacity, and decreased the proinflammatory cytokines levels in laying hens with Escherichia coli LPS challenge.

Effects of herbal cake-partitioned moxibustion on the expression of thyroid autophagy-related factors LC3B and Beclin-1 in rats with autoimmune thyroiditis

ObjectiveTo observe the anti-inflammatory effect, as well as the effect on the expression of microtubule-associated protein light chain 3B (LC3B) and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis (EAT).MethodsFemale Sprague-Dawley rats were randomly divided into a normal group and a modeling group. The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction. After the model was prepared, rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group. In the herbal cake-partitioned moxibustion group, moxibustion was alternately applied to two groups of points [Dazhui (GV14)-Mingmen (GV4) and Tiantu (CV22)-Guanyuan (CV4)], and the treatment continued for 30 d. Rats in the normal and model groups were only fixed identically without intervention. Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining; the concentrations of thyroid peroxidase antibodies (TPOAb), thyroglobulin antibodies (TGAb), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay; real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.ResultsThere were massive follicular destruction, lymphocytic infiltration, and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group. Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group. Compared with the normal group, the concentrations of serum TPOAb, TGAb, IL-1β, IL-6, and TNF-α were increased in the model rats (P<0.05); the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group (P<0.05). Compared with the model group, the concentrations of serum TPOAb, TGAb, IL-1β, IL-6, and TNF-α were reduced in the herbal cake-partitioned moxibustion group (P<0.05); the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group (P<0.05). The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.ConclusionHerbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb. This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.

Ornithine Aspartate and Vitamin-E Combination Has Beneficial Effects on Cardiovascular Risk Factors in an Animal Model of Nonalcoholic Fatty Liver Disease in Rats.

Cardiovascular (CV) disease is the main cause of death in nonalcoholic fatty liver disease (NAFLD), a clinical condition without any approved pharmacological therapy. Thus, we investigated the effects of ornithine aspartate (LOLA) and/or Vitamin E (VitE) on CV parameters in a steatohepatitis experimental model. Adult Sprague Dawley rats were randomly assigned (10 animals each) and treated from 16 to 28 weeks with gavage as follows: controls (standard diet plus distilled water (DW)), NAFLD (high-fat choline-deficient diet (HFCD) plus DW), NAFLD+LOLA (HFCD plus LOLA (200 mg/kg/day)), NAFLD+VitE (HFCD plus VitE (150 mg twice a week)) or NAFLD+LOLA+VitE in the same doses. Atherogenic ratios were higher in NAFLD when compared with NAFLD+LOLA+VitE and controls (p < 0.05). Serum concentration of IL-1β, IL-6, TNF-α, MCP-1, e-selectin, ICAM-1, and PAI-1 were not different in intervention groups and controls (p > 0.05). NAFLD+LOLA decreased miR-122, miR-33a, and miR-186 (p < 0.05, for all) in relation to NAFLD. NAFLD+LOLA+VitE decreased miR-122, miR-33a and miR-186, and increased miR-126 (p < 0.05, for all) in comparison to NAFLD and NAFLD+VitE. NAFLD+LOLA and NAFLD+LOLA+VitE prevented liver collagen deposition (p = 0.006) in comparison to NAFLD. Normal cardiac fibers (size and shape) were lower in NAFLD in relation to the others; and the inverse was reported for the percentage of regular hypertrophic cardiomyocytes. NAFLD+LOLA+VitE promoted a significant improvement in atherogenic dyslipidemia, liver fibrosis, and paracrine signaling of lipid metabolism and endothelial dysfunction. This association should be further explored in the treatment of NAFLD-associated CV risk factors.

Pyroptosis associated with immune reconstruction failure in HIV-1- infected patients receiving antiretroviral therapy: a cross-sectional study

BackgroundHighly active anti-retroviral therapy (HAART) can successfully suppress human immunodeficiency virus (HIV) viral replication and reconstruct immune function reconstruction in HIV-1-infected patients. However, about 15–30% of HIV-1-infected patients still fail to recover their CD4+ T cell counts after HAART treatment, which means immune reconstruction failure. Pyroptosis plays an important role in the death of CD4+ T cells in HIV-1- infected patients. The study aims to explore the association between the expression of pyroptosis in peripheral blood and immune function reconstruction in HIV-1- infected patients.MethodsOne hundred thirty-five HIV-1-infected patients including immunological non-responders (INR) group, immunological responders (IR) group and normal immune function control (NC) group were analyzed. The expression of GSDMD and Caspase-1 in peripheral blood of HIV-1-infected patients were measured by qPCR. The concentrations of GSDMD, Caspase-1, IL-1β and IL-18 in the peripheral serum were quantified by ELISA. The associations between the expression of pyroptosis in peripheral blood and immune function reconstruction were analyzed using multivariate logistic models.ResultsThe relative expression of GSDMD mRNA and caspase-1 mRNA in peripheral blood, as well as the expression of IL-18 cytokine in the INR, were significantly higher than those in the IR and NC (P < 0.05). There was no significant difference in the expression of IL-1β cytokine (P > 0.05). Multivariate logistic analysis showed that the patients with baseline CD4+ T cell counts less than 100 cells/μL (aOR 7.051, 95% CI 1.115–44.592, P = 0.038), high level of expression of Caspase-1mRNA (aOR 2.803, 95% CI 1.065–7.377, P = 0.037) and IL-18 cytokine (aOR 10.131, 95% CI 1.616–63.505, P = 0.013) had significant poor CD4+ T cell recovery.ConclusionsThe baseline CD4+ T cell counts less than 100 cells/μL, high relative expression of Caspase-1 mRNA, and high expression of IL-18 cytokine are associated factors that affect the reconstruction of immune function.