Expression Of IL-10 Genes Research Articles

Phosphatidylserine Topically Attenuates Imiquimod-induced Psoriasis Through Inflammation Inhibition in Mice.

Psoriasis is a chronic skin condition that is associated with persistent inflammation and skin lesions. Topical therapy has been a promising approach to the alleviation of psoriasis through the application of anti-inflammatory agents. Phosphatidylserine (PS) administration has shown anti-inflammatory effects in the trials. Consequently, the objective of this study was to evaluate the effects of topical PS on the potential improvement of an imiquimod (IMQ)-induced psoriasis model. Additionally, cyclosporine A was utilized as a comparative anti-psoriatic agent in our study. The psoriasis model was established by topically applying IMQ to the dorsal skin of mice once daily for five consecutive days. The efficacy of topical PS was assessed using the Psoriasis Area and Severity Index (PASI) score to evaluate skin lesions. Subsequently, the skin samples were analyzed using Baker's scoring system, Masson's trichrome staining, immunohistochemistry, and real-time PCR analysis. IMQ-induced plaque-type psoriasis resulted in a significant increase (P<0.05) in dermal thickness, hyperkeratosis, PASI score, and inflammatory cytokines at the lesion site. The topical PS and cyclosporine A significantly (P<0.05) reduced PASI score and dermal thickness, while also alleviating erythema and scaling when compared to untreated mice. Furthermore, biomolecular assessments revealed that PS significantly (P<0.05) inhibited the gene expression of IL-17, IL-23, and TNF-α cytokines in the IMQ-induced lesions. Topical PS may pointedly alleviate psoriasis through the inhibition of inflammation. The beneficial effects of the PS recommend further investigation in both experimental and clinical studies in the control of skin psoriasis.

Hydroxychloroquine-Mediated Autophagy Inhibition Shifts Monocyte-Derived Dendritic Cells Toward Immunogenic Phenotype In Vitro

Dendritic cells (DCs) are recognized as the most potent antigen-presenting cells (APCs) and act as intermediaries between the innate and adaptive immune systems. The maturation and function of DCs are significantly influenced by autophagy. Hence, the current research evaluated the effect of hydroxychloroquine (HCQ), an autophagy inhibitor, on the maturation and activation of monocyte-derived DCs. Monocytes were cultured and induced to differentiate into monocyte-derived DCs by the addition of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) cytokines. After five days, the immature DCs were exposed to HCQ and Lipopolysaccharides (LPS), followed by a 24-hour incubation period. On the sixth day, the flow cytometry technique was employed to assess the impact of HCQ on the phenotypic characteristics of the DCs. Finally, the expression of both pro- and anti-inflammatory genes in the HCQ-treated and untreated DCs groups was examined using the real-time PCR method. DCs that were subjected to treatment with HCQ exhibited a notable augmentation in the expression of CD11c, CD86, and HLA-DR, which are markers associated with maturation and antigen presentation. Furthermore, the DCs treated with HCQ demonstrated a significant reduction in the expression of IL-10, TGF-β, and IDO genes, while displaying an increase in the expression of TNF-α and IL-12 when compared to the control group. These findings indicate that targeting autophagy can induce a shift in DCs towards an immunogenic state. However, further investigations are necessary to assess the impact of HCQ-treated DCs on T cell responses both in vitro and in vivo in tumor-bearing animal models.

Phylogenetic position of the pigeon mite, Ornithonyssus sylviarum, with amplification of its immunogenetic biomarkers in Egypt

Ornithonyssus sylviarum (O. sylviarum) is an obligatory, blood-sucking ectoparasite widely distributed among poultry and other mammals, causing significant economic losses. This study represented the first report of molecular genotypic identification of O. sylviarum from pigeons, Columba livia domestica, in Egypt. PCR and sequencing of the 28S rRNA gene were conducted. The resulting mite sequences were subjected to BLAST analysis, revealing 90–100% similarity to O. sylviarum in all tested samples. The sequences were deposited in GenBank under the accession numbers PP049086 and PP033720. A phylogenetic tree was constructed to compare the obtained species with related species worldwide. Additionally, infected pigeons showed increased expression of IL-1, IL-10, IFN-γ, and TGF-β3 genes and elevated serum levels of stress biomarkers. The increased level of these cytokines indicates there was a disturbance in the immune status of the infected host with parasite compared with control healthy ones. This increases the susceptibility to infection with other pathogens.

The Indigenous Probiotic Lactococcus lactis PH3-05 Enhances the Growth, Digestive Physiology, and Gut Microbiota of the Tropical Gar (Atractosteus tropicus) Larvae.

Probiotics in aquaculture hold promise for enhancing fish health and growth. Due to their increased specificity and affinity for their host, indigenous probiotics may offer isolated and potentially amplified benefits. This study investigated the effects of Lactococcus lactis PH3-05, previously isolated from adults of tropical gar (Atractosteus tropicus), on the growth, survival, digestive enzyme activity, intestinal morphology, expression of barrier and immune genes, and intestinal microbiota composition in the larvae of tropical gar. Larvae were fed with live L. lactis PH3-05 concentrations of 104, 106, and 108 CFU/g for 15 days alongside a control diet without probiotics. Higher concentrations of L. lactis PH3-05 (106 and 108 CFU/g) positively influenced larval growth, increasing hepatocyte area and enterocyte height. The 106 CFU/g dose significantly enhanced survival (46%) and digestive enzyme activity. Notably, the 108 CFU/g dose stimulated increased expression of muc-2 and il-10 genes, suggesting enhanced mucosal barrier function and anti-inflammatory response. Although L. lactis PH3-05 did not significantly change the diversity, structure, or Phylum level composition of intestinal microbiota, which was constituted by Proteobacteria, Bacteroidota, Chloroflexi, and Firmicutes, an increase in Lactobacillus abundance was observed in fish fed with 106 CFU/g, suggesting enhanced probiotic colonization. These results demonstrate that administering L. lactis PH3-05 at 106 CFU/g promotes growth, survival, and digestive health in A. tropicus larvae, establishing it as a promising indigenous probiotic candidate for aquaculture applications.

Elucidating the effect of dietary neem (Azadirachta indica) on growth performance, haemato-biochemical, immunonological response, and anti-pathogenic capacity of Nile tilapia juveniles.

This investigation attempts to evaluate the effect of diet additives via aqueous or ethanolic herbal extracts from Azadirachta indica leaves on Nile tilapia, Oreochromis niloticus. Five dietary categories were assigned to the fish: the first category (N1, with no extract) was kept under control conditions; two categories contained aqueous extract (N2 (1.0g/kg) and N3 (2.0g/kg); and two categories contained ethanolic extract, N4 (1.0g/kg) and N5 (2.0g/kg), with each group being fed for 60 days. After the feeding trial, Aeromonas hydrophila was injected intraperitoneally into fish for 14 days; fish mortality was recorded during this period. The results showed that the fish-fed dietary A. indica significantly improved growth performance and intestinal health (digestive enzymes and intestinal morphology), especially in the N4 and N5 categories. However, N4 and N5 categories demonstrated a significant decrease in AST and ALT activities and an increase in total protein, serum albumin, globulin, growth hormone (GH), leptin hormone (LEP), hemoglobin, white blood cells, and hematocrit (P < 0.05) in comparison with the control category (N1). Compared to the control category, the N4 and N5 categories have revealed a significant reduction in MDA activity and improvements in immunological activities (lysozyme, complement C3, and nitric oxide) and antioxidant enzymes (CAT, SOD, and GPX). Moreover, in tilapia-fed A. indica, the expression of IL-8, IL-1β, and Nf-κb genes was downregulated partially in the N4 and N5 categories than the control category. In contrast, the lysozyme, C3, GPX, and CAT genes were upregulated partially at N4 and N5 compared to the control category. Following the bacterial challenge, fish in the N4 and N5 categories also displayed the lowest fish mortality compared to the control category. The ethanolic extract displayed a more potent resistance against the parasite Cichlidogyrus tilapia in vitro than the aqueous and control categories, partially at 2g/L. According to these findings, an ethanolic neem extract (2.0g/kg feed) activates the immune system and antioxidant response in Nile tilapia fingerlings, improving growth and fish resistance to parasitic and bacterial infections.

Dietary Psidium guajava, guava leaf extract protects Oreochromis niloticus, Nile tilapia from Pseudomonas aeruginosa infection and enhances growth

This study investigated the impact of dietary guava leaf extract (GLE) on immunity, resistance to Pseudomonas aeruginosa infection, and growth in Oreochromis niloticus (Nile tilapia). Four groups of fish (28.6 g.fish−1) were fed diets supplemented with ethanolic GLE at 0 (control), 3, 5, or 7 g.fish−1 diet (GLE0diet, GLE3diet, GLE5diet, GLE7diet) at 28 °C. Fish were fed thrice daily at 15 g.kg−0.8.d−1 for 42 days. Innate immune parameters, including mucus-bactericidal activity (MBA), serum-bactericidal activity (SBA), bacterial agglutination activity (BAA), and phagocytic activity (PA), were assessed on days 14, 28, and 42. Weight gain (WG), specific growth rate (SGR), and feed conversion ratio (FCR) were measured on day 42. Post-feeding, fish were injected with P. aeruginosa, and mortality was recorded for 30 days. Results showed that GLE7diet significantly increased MBA, SBA, BAA, and PA by 1.8-, 3.3-, 4.2-, and 3.5-fold, respectively, compared to control. GLE7diet also showed the highest significant reduction in post-challenge mortality (21 %) compared to the control group (83 %). Growth metrics for GLE7diet showed WG of 43.8 g, SGR of 2.21%.d−1, and FCR of 1.19, compared to WG of 37.1 g, SGR of 1.98%.d−1, and FCR of 1.41 in the control group. SGR showed an exponential increase (R² = 0.935) and FCR a decrease (R² = 0.958) with increasing GLE levels. GLE5diet and GLE7diet significantly increased MBA at all-time points compared to control. GLE7diet significantly increased SBA on days 14, 28, and 42 compared to control. GLE7diet significantly enhanced BAA at all-time points. GLE5diet and GLE7diet significantly increased PA at all time points compared to control. GLE7diet led to increased expression of IL-1, IL-6, IFN-α, IFN-β, and TNF-α genes, particularly from 10 to 30 days post-infection. Dietary GLE at 7 g.kg−1 significantly enhanced innate and acquired immune responses, growth performance, and resistance to P. aeruginosa in Nile tilapia. This study recommended incorporating 7 g.kg−1 GLE in Nile tilapia diets to improve disease resistance and growth performance.

Dermal derived matrix hydrogel loaded with curcumin improved wound healing in a diabetic rat model

There is a need in clinical practice for new wound healing techniques to address full thickness skin injuries, particularly in individuals with diabetes. Herein we investigated whether dermal derived matrix hydrogel (DMH) loaded with curcumin (Cur) could promote healing in diabetic rats. Sixty diabetic rats were randomly assigned into the non-treated group, DMH group, Cur group, and DMH+Cur group. According to the phases of wound healing, sampling was done on days 7, 14, and 21 for further assessments. Our results indicated that the wound contraction rate, new epidermal length and thickness, number of fibroblasts and vascular length, collagen deposition, and strength properties of the healed wounds were meaningfully increased in the treatment groups than in the non-treated group, and these changes were more obvious in the DMH+Cur ones. In addition, the expression of VEGF and IL-10 genes were meaningfully upregulated in all treatment groups compared to the non-treated group and were greater in the DMH+Cur group. This is while the number of neutrophils and expression levels of TNF-α and IL-1β genes decreased more significantly in the DMH+Cur group compared to the other groups. In conclusion, it was found that using both DMH and curcumin has a greater impact on diabetic wound healing.

Effect of Lactiplantibacillus plantarum on the growth, hemato-biochemical, inflammation, apoptosis, oxidative stress markers, involved gens and histopathological alterations in growing rabbits challenged with aflatoxin B1

Aflatoxin B1 (AFB1) is a significant pollutant found in food and feed, posing a threat to public health. The objective of this study was to assess the effect of Lactiplantibacillus plantarum (LACP) against AFB1 in growing rabbits by investigating growth, serum metabolites, immunity, antioxidant capacity, and inflammatory response. A total of 60 growing male rabbits (721.5 ± 2.68g) were allocated to 4 experimental groups. The control group receiving only a basal diet, the AFB1 group (0.3 mg AFB1/kg diet), the LACP group (1 × 109 cfu/g /kg diet), and the combination group (1 × 109 cfu/g + 0.3 mg AFB1/kg diet; AFB1+ LACP) for 8 wk. The administration of AFB1 alone significantly decreased the final body weight, body gain, and feed intake, while significantly increasing the feed conversion ratio (P < 0.05). A significant decline in total proteins and globulins, along with elevated levels of hepatic enzymes (AST, ALP, ALT, and GGT) and renal function markers (creatinine and uric acid), were observed in the AFB1-contaminated group (P < 0.05). Immunoglobulins (IgG and IgM) were significantly decreased, alongside a significant elevation of triglycerides, direct bilirubin, and indirect bilirubin in growing rabbits fed diets with AFB1 (P < 0.05). Supplementing the AFB1 diet with LACP restored the growth reduction, improved liver (AST, ALP, ALT, and GGT) and kidney (creatinine and uric acid) functions, and enhanced immune markers in rabbit serum (P < 0.05). Antioxidant indices (SOD, GSH, and CAT) were significantly decreased in the AFB1 group (P < 0.05). However, the addition of LACP to the AFB1-contaminated diets improved antioxidant capacity and malondialdehyde (MDA) and protein carbonylation (PC) in hepatic tissues of rabbits (P < 0.05). Serum interlukin-4 (IL-4) and interferon gamma (IFN-γ) levels were significantly increased in the AFB1 group (P < 0.05), but the addition of LACP significantly reversed this elevation. AFB1 downregulated the expression of immune-inflammatory genes such Nrf2, IL-10, and BCL-2 genes, while up-regulating the caspase-3 (CASP3) gene (P < 0.05). Supplementing AFB1 diet with LACP significantly decreased the expression of immune-inflammatory genes (Nrf2, IL-10, and BCL-2) and reduced the expression of the apoptotic-related gene CASP3. This study highlights the potential of L. plantarum (1 × 109 cfu/g /kg diet) as a protective agent against AFB1 in growing rabbits by enhancing antioxidant and immune function and reducing apoptosis and inflammation pathways.

The effects of Artemisia Sieberi, Achillea Fragrantissima, and Olea Europaea leaves on the performance and physiological parameters in heat-stressed broiler chickens.

High temperatures have detrimental effects on the performance and physiology of broiler chickens. Medicinal plants have various biological activities and may enhance the heat resistance of chickens during heat waves. Therefore, this study aimed to explore the potential roles of using specific local medicinal plants to alleviate the negative impacts of heat stress (HS) in broilers. In this study, 180 day-old chicks were used to investigate the effects of HS and dietary indigenous medicinal plants on growth performance, antioxidant biomarkers, and intestinal health. The chicks were assigned to six groups (18 pens with 10 chicks per pen) with three replicates each. In the first group, the chicks were kept under thermoneutral conditions (CON) and fed a basal diet. The other five groups were exposed to recurrent heat stress and fed a basal diet (T1, HS group) or supplemented with Artemisia Sieberi (1.25 g/kg of feed; T2), Achillea Fragrantissima (15 g/kg of feed; T3), Olea europaea (10 g/kg of feed; T4), and all the previous additives (all-in-one) combined at the same dose levels mentioned above (T5). At 21 days of age, the chicks from each group were exposed to two phases of heat stress: phase 1 from days 21 to 34 (34 ± 1°C) followed by phase 2 from days 35 to 39 (37 ± 1°C). The results indicate that HS significantly increased rectal temperature and respiration rate in broiler chickens. Feed intake and body weight gain were improved in all supplemented groups, while the feed conversion ratio was decreased in response to the dietary inclusion of medicinal plants. Additionally, glutathione peroxidase and immunoglobulin G levels were increased in the T3, T4, and T5 groups compared to the other groups. HS induced significant upregulated in the mRNA levels of heat shock protein 70 and interleukin-8, while the mRNA of occludin was decreased. The T3, T4, and T5 showed significantly decreased expression of hepatic HSP70 and ileum IL-8 genes and increased ileum mRNA occludin levels relative to the CON and T1 groups. In conclusion, supplementation with these plants enhances growth performance and maintains intestinal health sustaining the productivity of broiler chickens under HS conditions.

Comparison of serum levels of IL-10 and IL-11 and mRNA expression of IL-10, IL-11, COX-2, BCL6, and ZEB Family in peripheral blood mononuclear cells (PBMC) of women with polycystic ovary syndrome and healthy individuals

BackgroundThe roles of IL-10, IL-11, COX-2, BCL6, ZEB1, and ZEB2 genes in the potential correlation between polycystic ovary syndrome (PCOS), inflammation, and cancer remain controversial. AimsThis study aimed to compare serum levels of IL-10 and IL-11 and gene expression of IL-10, IL-11, COX-2, BCL6, ZEB1, and ZEB2 in PBMCs of women with PCOS and healthy controls. MethodsA case-control study included 40 women with PCOS as the case group and 40 healthy women as controls. Group matching for age and BMI was performed. Serum levels of IL-10 and IL-11 were assessed using ELISA, while gene expression was measured using real-time PCR. Parameters were compared between groups, and correlations among gene expression and serum levels were explored. ResultsIn comparison to healthy women, women with PCOS exhibited a significant decrease in the expression of COX-2 and IL-10 genes (p<0.001), alongside a significant increase in ZEB2 gene expression (p<0.001). There were no significant differences observed in the expression of IL-11, BCL6, and ZEB1 genes. Furthermore, the serum level of IL-10 was significantly lower in women with PCOS compared to the control group (p<0.001), while no significant difference was found in IL-11 levels. Additionally, no significant correlations were identified between gene expression and serum levels. ConclusionIn women with PCOS, reduced IL-10 gene expression may indicate inflammation and serve as a diagnostic biomarker. However, conflicting findings on COX-2 expression complicate understanding. Elevated ZEB2 expression in PCOS women may lead to infertility, epithelial-mesenchymal transition, and aggressive phenotypes.

Microparticles Incorporating Dual Apoptotic Factors to Inhibit Inflammatory Effects in Macrophages

New approaches to treat autoimmune diseases are needed, and we can be inspired by mechanisms in immune tolerance to guide the design of these approaches. Efferocytosis, the process of phagocyte-mediated apoptotic cell (AC) disposal, represents a potent tolerogenic mechanism that we could draw inspiration from to restore immune tolerance to specific autoantigens. ACs engage multiple avenues of the immune response to redirect aberrant immune responses. Two such avenues are: phosphatidylserine on the outer leaflet of the cell and engaging the aryl hydrocarbon receptor (AhR) pathway. We incorporated these two avenues into one acetalated dextran (Ace-DEX) microparticle (MP) for evaluation in vitro. First phosphatidylserine (PS) was incorporated into Ace-DEX MPs and evaluated for cellular association and mediators of cell tolerance including IL-10 production and M2 associated gene expression when particles were cultured with peritoneal macrophages (PMacs). Further PS Ace-DEX MPs were evaluated as an agent to suppress LPS stimulated PMacs. Then, AhR agonist 2-(1′H-indole-3′-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE) was incorporated into Ace-DEX MPs and expression of M2 and IL-10 genes was evaluated in PMacs. Further the ITE and PS Ace-DEX MPs (PS/ITE MPs) were evaluated for suppression of T cell priming and Th1 polarization. Our results indicate that the PS/ITE-MPs stimulated anti-inflammatory cytokine expression and suppressed inflammation following LPS stimulation of PMacs. Moreover, PS/ITE MPs induced the anti-inflammatory enzyme IDO1 and suppressed macrophage-mediated T cell priming and Th1 polarization. These findings suggest that PS and ITE-loaded Ace-DEX MPs could be a promising therapeutic tool for suppressing inflammation.

Ganoderma lucidum dry extract supplementation modulates T lymphocyte function in older women.

Ganoderma lucidum (a mushroom used in traditional Chinese medicine) compounds may attenuate ageing-related physiological changes and restore normal immunity. However, studies on the physiological effects of Ganoderma lucidum dry extract food supplements are few. Therefore, here, we aimed to investigate the effects of Ganoderma lucidum dry extract food supplement on the lymphocyte function of older women. This was a double-blind clinical trial (n 60) with a final 39 older volunteers, divided into two groups Ganoderma lucidum (n 23) and placebo (n 16). The Ganoderma lucidum group received 2000 mg/d of Ganoderma lucidum dry extract for 8 weeks. We used flow cytometry to determine the lymphocyte profile. CD4+ lymphocyte gene expression was evaluated by real-time polymerase chain reaction. We observed that in the Ganoderma lucidum group, concanavalin A stimulation increased lymphocyte proliferation. Further, we observed an increase in expression of Forkhead box P3, transforming growth factor-beta, IL-10, IL-6, retinoic acid receptor-related orphan receptor gamma, GATA-binding protein 3 and interferon gamma genes in the Ganoderma lucidum group. Furthermore, in the Ganoderma lucidum group, ionomycin and phorbol 12-myristate 13-acetate stimulation led to decrease in Th17+ cells and increase in Th2+ cells. Thus, in older women, Ganoderma lucidum regulates T lymphocyte function leading to a predominant anti-inflammatory action but does not induce T lymphocyte proliferation through CD28 signalling pathway.

Hydatid fluid from Echinococcus granulosus induces autophagy in dendritic cells and promotes polyfunctional T-cell responses.

Parasites possess remarkable abilities to evade and manipulate the immune response of their hosts. Echinococcus granulosus is a parasitic tapeworm that causes cystic echinococcosis in animals and humans. The hydatid fluid released by the parasite is known to contain various immunomodulatory components that manipulate host´s defense mechanism. In this study, we focused on understanding the effect of hydatid fluid on dendritic cells and its impact on autophagy induction and subsequent T cell responses. Initially, we observed a marked downregulation of two C-type lectin receptors in the cell membrane, CLEC9A and CD205 and an increase in lysosomal activity, suggesting an active cellular response to hydatid fluid. Subsequently, we visualized ultrastructural changes in stimulated dendritic cells, revealing the presence of macroautophagy, characterized by the formation of autophagosomes, phagophores, and phagolysosomes in the cell cytoplasm. To further elucidate the underlying molecular mechanisms involved in hydatid fluid-induced autophagy, we analyzed the expression of autophagy-related genes in stimulated dendritic cells. Our results demonstrated a significant upregulation of beclin-1, atg16l1 and atg12, indicating the induction of autophagy machinery in response to hydatid fluid exposure. Additionally, using confocal microscopy, we observed an accumulation of LC3 in dendritic cell autophagosomes, confirming the activation of this catabolic pathway associated with antigen presentation. Finally, to evaluate the functional consequences of hydatid fluid-induced autophagy in DCs, we evaluated cytokine transcription in the splenocytes. Remarkably, a robust polyfunctional T cell response, with inhibition of Th2 profile, is characterized by an increase in the expression of il-6, il-10, il-12, tnf-α, ifn-γ and tgf-β genes. These findings suggest that hydatid fluid-induced autophagy in dendritic cells plays a crucial role in shaping the subsequent T cell responses, which is important for a better understanding of host-parasite interactions in cystic echinococcosis.

Comparison of the recovery characteristics for canine corneal ulcer treated with corneoconjunctival transposition or conjunctival autografts

Corneal ulceration induced by Staphylococcus pseudintermedius (S. pseudintermedius) is a common clinical eye disease. Antibiotics combined with corneoconjunctival transposition (CCT) or conjunctival autografts (CA) are often used, but the recovery characteristics are still unknown. In this experiment, canine corneal ulcer models induced by S. pseudintermedius and treated with levofloxacin eye drops (LED) were created. The models were used to compare the recovery characteristics of CCT and CA, combined with LED, by clinical observation, histopathology, and cytokine expression detected by qRT-PCR analysis. The results showed that the ulcerative cornea with only LED treatment perforated after 48 h. The mRNA expression of TLR2, IL-1β, IL-6, IL-8, and TNF-α genes was significantly elevated on 14, 28, and 35 days after the surgery compared to normal (p < 0.01). On day 42, the inflammatory damage had resolved, but the corneal transparency and arrangement of collagen fibrils in the CCT group were higher than those in the CA group. The mRNA expression of EGF, FGF, TGF-β1 and VEGF genes increased significantly (p < 0.01), mostly until day 42, proving that CCT and CA surgery contributed to the corneal recovery, and relieved the inflammatory reaction, with the elimination of corneal cicatrices needing a period of reconstruction. Therefore, this study has provided, for the first time, the method for establishing a canine corneal ulcer model induced by S. pseudintermedius. More importantly, the recovery of canine ulcerative corneas with CCT or CA surgery is reported for the first time.

Biologic Adjuvants to Rotator Cuff Repairs Induce Anti-inflammatory Macrophage 2 Polarization and Reduce Inflammatory Macrophage 1 Polarization In Vitro

PurposeTo examine the effect of various biologic adjuvants on the polarization of macrophages in an in vitro model for rotator cuff tears. MethodsTissue was harvested from 6 patients undergoing arthroscopic rotator cuff repair. An in vitro model of the supraspinatus and subacromial bursa was created and treated with control, platelet rich plasma (PRP), autologous activated serum (AAS), or a combination of PRP+AAS. The effect of treatment on macrophage polarization between M1 pro-inflammatory macrophages or M2 anti-inflammatory macrophages was measured using gene expression, protein expression, flow cytometry and nitric oxide (NO) production. ResultsTendon and bursa treated with PRP, AAS and PRP+AAS significantly decreased the gene expression of M1 markers IL-12 and TNF-a, while significantly increasing the expression of M2 markers Arginase, IL-10 and TGF-b (p<0.05) compared to treatment with control. ELISA analysis of protein production demonstrated that compared to control, co-culture treated with PRP, AAS and PRP+AAS significantly decreased markers of M1-macrophages (IL-6, IL-12, and TNF-a), while significantly increasing the expression of markers of M2-macrophages (Arginase, IL-10, and TGF-b) (p<0.05). Flow cytometry analysis of surface markers demonstrated that compared to control, tendon and bursa treated with PRP, AAS and PRP+AAS significantly decreased markers of M1-macrophages (CD80, CD86, CD64, CD16), while significantly increasing the expression of markers of M2-macrophages (CD163 and CD206) (p<0.05). Treatment of the co-culture with PRP, AAS and PRP+AAS consistently demonstrated a decrease in NO production (p<0.05) compared to control. AAS and PRP+AAS demonstrated an increased macrophage shift to M2 compared to PRP alone, whereas there was not as uniform of a shift when comparing PRP+AAS to AAS alone. ConclusionsIn an in vitro model of rotator cuff tears, the treatment of supraspinatus tendon and subacromial bursa with PRP, AAS and PRP+AAS demonstrated an increase in markers of anti-inflammatory M2-macrophages and a concomitant decrease in markers of pro-inflammatory M1-macrophages. AAS and PRP+AAS contributed to a large shift to macrophage polarization to the anti-inflammatory M2 compared to PRP.

Innate immune response of snakehead fish to Indian strain of snakehead rhabdovirus (SHRV-In) infection and the infectivity potential of the virus to other freshwater fishes

A new virus known as snakehead rhabdovirus (SHRV-In) was discovered in South India in striped snakehead (Channa striata) that had hemorrhagic patches and cutaneous ulcerations. The virus is the most potentially harmful pathogen of snakehead because it could cause 100% mortality within 5 days. The goal of the current investigation was to evaluate the infectivity of rhabdovirus in freshwater fishes and to analyze the immune response in snakehead fish after challenge with SHRV-In. The infectivity study of SHRV-In against three freshwater fish such as tilapia, grass carp and loach showed that the virus could not induce mortality in any of them. Snakehead fish challenged with SHRV-In showed significant (p < 0.05) changes in haematological parameters such as red blood cell (RBC), haemoglobin (HGB), haematocrit (HCT), mean corpuscular haemoglobin concentration (MCHC), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), white blood cell (WBC), total platelet (PLT) counts, mean platelet volume (MPV) and immunological markers such as respiratory burst, superoxide dismutase, catalase activity and myeloperoxidase activity at 6, 12, 24 and 48 hpi. Real time PCR was executed to examine the expression profile of innate immune genes such as IRF-7, IL-8 and IL-12 in Snakehead fish at 6, 12, 24 and 48 h post SHRV-In infection. Immune gene expression of IRF-7, IL-8 and IL-12 were up-regulated in the spleen when compared to kidney at 6 and 12 hpi. However, the expression level of all the genes was down-regulated at 24 and 48 hpi. The down regulation of innate immune genes after 24 hpi in these tissues may be the result of increased multiplication of SHRV-In by interfering with the immune signaling pathway.

Effects of tamoxifen on the immune response phenotype in equine peripheral blood mononuclear cells.

Tamoxifen (TAM) is widely utilized in the prevention and treatment of human breast cancer and has demonstrated the potential to modulate the immune response. It has been proposed as a therapeutic tool for immune-mediated diseases. TAM has been investigated as a possible treatment for asthma-like conditions in horses, revealing specific impacts on the innate immune system. While the effects of TAM on equine neutrophils are well-documented, its influence on lymphocytes and the modulation of the immune response polarization remains unclear. This in vitro study employed peripheral blood mononuclear cells (PBMC) from healthy horses, exposing them to varying concentrations of the TAM and assessing the expression of genes involved in the polarization of the immune response (TBX21, IFNG, GATA3, IL4, IL10, FOXP3, and CTLA4) in PBMC stimulated or not with PMA/ionomycin. Additionally, the effect of TAM over the proportion of regulatory T cells (Treg) was also assessed. TAM did not significantly affect the expression of these genes and Treg at low concentrations. However, at the highest concentration, there was an impact on the expression of GATA3, IL4, IL10, and CTLA4 genes. These alterations in genes associated with a Th2 and regulatory response coincided with a noteworthy increase in drug-associated cytotoxicity but only at concentrations far beyond those achieved in pharmacological therapy. These findings suggest that the effects of TAM, as described in preclinical studies on asthmatic horses, may not be attributed to the modification of the adaptive response.

Promising effects of 1,8 Cineole to control Giardia lamblia infection: Targeting the inflammation, oxidative stress, and infectivity

Reportedly, synthetic drugs such as metronidazole, furazolidone, tinidazole, and quinacrine are used for the treatment of giardiasis but are associated with adverse effects. In this study, we aimed to investigate the in vitro and in vivo effects of eucalyptol (ECT, 1,8 cineole) alone and in combination with metronidazole (MNZ) on Giardia lamblia. The effects of ECT on cell viability, plasma membrane permeability, and gene expression levels of adenylate cyclase (AK) and extracellular signal kinases 1 and 2 (ERK1 and ERK2) in trophozoites of G. lamblia were assessed. In vivo, the effects of ECT alone and in combination with MNZ were assessed on mice infected with G. lamblia. In addition, the gene expression of inflammatory genes (e.g., TNF-α, IL-1β, and IL-10) and antioxidant genes (catalase (CAT), superoxide dismutase 1 (SOD1), glutathione peroxidase 2 (GPX2)) was determined by real-time PCR. The IC50 values of ECT, MNZ, and ECT+MNZ on trophozoites were 30.2 µg/mL, 21.6 µg/mL, and 8.5 µg/mL, respectively. The estimated Fractional inhibitory concentration index (FICI) values for ECT and MNZ were 0.28 and 0.39, respectively. The application of ECT on G. lamblia trophozoites resulted in a dose-dependent increase in plasma membrane permeability, particularly at concentrations of ½ IC50 and IC50 (P < 0.05). The treatment of infected mice with various doses of ECT, mainly in combination with MNZ for 7 days, resulted in a significant decrease (P < 0.001) in the average number and viability of cysts. ECT, especially when combined with MNZ, caused a significant (P < 0.001) reduction in the expression of TNF-α and IL-6 genes, and an increase (P < 0.05) in the expression of IL-10 genes. ECT alone and mainly in combination with MNZ leads to a significant (P < 0.001) increase in the gene expression of CAT, SOD, and GPX genes. These findings demonstrate that the use of ECT in these doses, even for 14 days, does not have any toxic effects on the function of vital liver and kidney tissues. The study findings confirmed the promising effects of ECT against G. lamblia infection both in vitro and in vivo. Considering the possible mechanisms, ECT increases plasma membrane permeability and reduces the expression levels of infectivity-related genes. In addition, ECT suppresses inflammation and oxidative stress, controlling giardiasis in mice. More studies are needed to clarify these findings.

Exploring the sex dimorphism in the expression of intestinal barrier and immune-related genes and intestinal microbiota in cage-cultured large yellow croaker (Larimichthys crocea) during the overwintering period along the Zhoushan coast

In Zhejiang province, large yellow croakers are primarily cultured in net cages, facing significant challenges during the overwintering period such as susceptibility to cold and starvation stress. Notably, the observable sexual dimorphism in the large yellow croaker hints at the likelihood of gender differences in their responses to these environmental stresses. However, the potential sex-specific adaptive changes during overwintering remain unexplored. To gain deeper insights, we investigated the expression of intestinal barrier-related genes, immune responses, and changes in intestinal microbiota during the overwintering period in males and females separately. The results revealed a more pronounced loss of body weight in females than that in males. In male intestines, there was a significant decrease in the expression of intestinal barrier-related genes (arp2/3, occludin, and zo1), contrasting with a significant increase in females. The expression of TLR1, TLR3, TLR7, TLR9, MyD88, and NF-κB genes in the intestines of female fish decreased significantly in March compared to November, while the opposite trend was observed in male fish. However, in the liver, TLR1, TLR3, TLR7, TLR9, MyD88, and NF-κB genes expression were both decreased significantly in males and females. In the male intestines, there was a significant increase in the expression of inflammatory cytokine genes (IL-1β and IL-6). In the females, IL-1β gene expression significantly decreased, while IL-6 expression increased significantly. The expression of IL-10 genes decreased in both males and females. In the liver, both the males and females exhibited a significant increase in the expression of IL-1β and IL-6 genes. Further analysis revealed greater susceptibility of male intestinal microbiota diversity during the overwintering period. Firmicutes’ relative abundance exhibited opposing changes between the males and females, and Proteobacteria abundance, driven by a significant increase in Vibrio bacteria, significantly increased in the males. In conclusion, the overwintering period may compromise the structural integrity of male fish intestines, reducing their immune function. Additionally, the response strategy of the intestinal microbiota differs between sexes. The findings provide crucial insights for crafting effective strategies and management decisions in cage-cultured large yellow croaker during the overwintering period, as well as offering theoretical references for monosex aquaculture.

Significant beneficial effects of 12-weeks add-on yoga therapy on antipsychotic-stabilized schizophrenia patients through epigenetic modulation: novel findings from a randomized controlled study

IntroductionComplementary and alternative therapy, especially yoga, is emerging as an important treatment modality for various complex disorders. Yoga therapy has reportedly been demonstrated to exhibit clinical benefits in schizophrenia. However, the modulatory effects of yoga therapy on the pathobiological pathways of schizophrenia are inadequately explored. Immune dysregulation is a widely recognized etiopathological construct of schizophrenia. It is not precisely known whether yoga therapy can modulate the expression of immune molecules by regulating gene expression and epigenetic processes in schizophrenia.ObjectivesTo understand the impact of 12-weeks add-on yoga therapy on the immune-inflammatory pathway in schizophrenia by examining plasma levels and gene expression levels of cytokines and complement proteins as well as by profiling promoter DNA methylation pattern of genes coding for cytokines and complement proteins.MethodsFifty-seven schizophrenia patients fulfilling DSM-V criteria were recruited into the study and randomized into Yoga therapy (n=28) and waitlist control (n=29) groups. Plasma levels of IL-1β, IL-6, IL-10, IL-17, C1q, C2, C3, C4, C5, C5a, Factor B and Factor H by Multiplex Suspension Assay, quantification of gene expression of Il1b, Il6, Il10, IL17, C3, C4 and C5 genes by quantitative PCR and promoter DNA methylation of Il1b, Il6, Il10, Il17, C3, C4 and C5 genes by pyrosequencing were carried out in all the study participants.ResultsPlasma levels of IL-1β (Z score= 2.42, p=0.02) dropped significantly and C2 (Z score= 2.24, p=0.03) levels increased after 12-weeks of yoga therapy. The expression of Il1b (Z score=2.45, p=0.01) and Il6 (Z score=2.07, p=0.04) genes were significantly downregulated, while the levels of C4 (Z score=2.23, p=0.03) gene was upregulated in schizophrenia patients of yoga therapy group. Two CpG sites in the promoter region of Il1b (all p≤0.05) and Il6 (all p≤0.05) genes and three CpG sites in the promoter region of C4 (all p&lt;0.05) gene were hypermethylated, while two CpG sites in the gene body of Il6 (all p≤0.05) gene and two CpG sites in the promoter region of Il10 (all p ≤0.05) gene were hypomethylated after 12-weeks of yoga therapy in schizophrenia patients.ConclusionsOur findings provide important insights into the mode of action of yoga therapy in schizophrenia. This study for the first time reports the epigenetic effects of yoga therapy on immune-inflammatory pathway in schizophrenia.Disclosure of InterestNone Declared