Main Cytokines Research Articles

Antioxidant and Anti-Inflammatory Activities of Stellera chamaejasme L. Roots and Aerial Parts Extracts.

Natural products, mainly plants, have a crucial role in folk medicine. Particularly, Stellera chamaejasme L. has been traditionally used in Mongolian medicine to treat various diseases, including chronic tracheitis, tuberculosis, and psoriasis. In this study, ethanol (EtOH) and dichloromethane (DCM) extracts of its roots (R) and aerial parts (AP) were evaluated for their antioxidant and anti-inflammatory activities. Thin-layer chromatography demonstrated the presence of flavonoids, namely kaempferol and quercetin-3-O-glucopyranoside, only in the EtOH-AP. Conversely, it showed that kaempferol, quercetin-3-O-glucopyranoside, coumarin, luteolin, rutin, morin, and riboflavin were not present in the other three extracts. The S. chamaejasme extracts exhibited strong antioxidant activity. In addition, the roots extracts presented the highest antioxidant activity against peroxyl radicals, with the EtOH-R being the most potent (IC50 = 0.90 ± 0.07 µg/mL). S. chamaejasme extracts also efficiently inhibited the production of one of the main pro-inflammatory cytokines, interleukin (IL)-6, in a dose-dependent manner by lipopolysaccharide-stimulated macrophages. Particularly, DCM-R was the strongest extract, reducing ≈ 91.5% of the IL-6 production. Since this extract was the most effective, gas chromatography-mass spectrometry (GC-MS) analyses were performed and demonstrated the presence of two fatty acids (palmitic acid and 9-octadecenoic acid), one fatty alcohol (1-hexadecanol), and one triterpenoid (squalene) that can contribute to the observed bioactivity. Herewith, S. chamaejasme extracts, mainly DCM-R, exhibit antioxidant and anti-inflammatory activities that could be applied as new and innovative natural formulations for the treatment of chronic inflammatory diseases.

T-lymphocytes FoxP3+ and their interconnection with the severity of coronary atherosclerosis in patients with coronary artery disease and diabetes mellitus type 2: a pilot study

BACKGROUND: One of the common pathogenic links of diabetes mellitus type 2 (T2DM) and coronary artery disease (CAD) is chronic low-grade inflammation, restricted by FoxP3+ T-regulatory lymphocytes.AIM: To investigate the numbers of FoxP3+CD25hi and FoxP3+CD25lo T-lymphocytes, the subcellular localization of FoxP3 in them, and the production of the main cytokines in relation to clinical and metabolic parameters in patients with association of CAD and T2DM.MATERIALS AND METHODS: An observational single-center single-stage comparative study was conducted. The severity of atherosclerosis was assessed by calculating the Gensini Score index after coronary angiography. Absolute numbers and frequencies of CD4+CD25hiFoxP3+ and CD4+CD25loFoxP3+ T-lymphocytes were assessed in peripheral blood by flow cytometry. Imaging flow cytometry was used to determine the degree of FoxP3 translocation to the cell’s nucleus. Concentration of cytokines in blood serum and supernatants of mononuclear leukocytes’ cultures was determined by the multiplex analysis.RESULTS: We recruited 57 patients with chronic CAD. Of these, T2DM was diagnosed in 22 patients. In patients with CAD and T2DM, the absolute numbers and frequencies of FoxP3+CD25lo cells were increased compared to patients with CAD without diabetes (1.15 (0.98; 1.73) vs. 0.96 (0.60; 1.15)% (р=0.046); 1.48 (1.05; 1.97) vs. 1.07 (0.71; 1.42) x107/L (р=0.025)). Patients with T2DM also had a higher level of translocation of FoxP3 to the nucleus of FoxP3+CD25lo cells (92.0 (86.4; 95.0) vs. 88.7 (80.0; 91.4)%, р=0.040) and increased concentration of the chemokine CCL22 both in blood serum (912 (828; 1061) vs. 669 (585; 738) pg/mL, р=0.022) and supernatants of LPS-stimulated mononuclear leukocyte cultures (1189 (851; 1310) vs. 539 (437; 949) pg/mL, р=0.038), which correlated with the presence of CD4+CD25hiFoxP3+ cells (Rs=0.587; p=0.044) and the triglyceride/glucose index (Rs=0.587; p=0.044). The identified changes were most pronounced in patients with moderately elevated values on the Gensini Score (17–45 points).CONCLUSION: We are the first to show association between the numbers of FoxP3+CD25lo-lymphocytes in peripheral blood and an increase in the nuclear translocation of FoxP3 in them with the severity of atherosclerosis in patients with association of CAD and T2DM. These data justify the necessity of the further investigation of the diagnostic significance of FoxP3+CD25lo-cells as biomarkers of tissue inflammation.

Non-alcoholic fatty liver disease as a risk factor for anemia of chronic inflammation (experimental research)

The aim of the study. In recent years, non-alcoholic fatty liver disease (NAFLD) has been considered a hepatic manifestation of the metabolic syndrome. The main consequence of NAFLD is chronic hepatic inflammation, which leads to dyslipidemia, inflammation, increased oxidative stress, and endothelial dysfunction. Immune activation in response to interaction with agents of a metabolic nature induces the release of pro-inflammatory cytokines in the liver, which subsequently cause iron сhomeostasis disorder. This leads to a frequent association of NAFLD with anemia of various etiology. In this regard, we considered it important to assess the severity of the systemic inflammatory response in NAFLD in the experiment in order to -diagnose anemia of chronic inflammation.Materials and methods. The study was carried out on 26 male Wistar rats, which were divided into control and experimental groups. In animals of the experimental group, NAFLD was modeled according to the generally accepted method. In order to assess metabolic disorders, we determined the main biochemical parameters, a complete blood count with the calculation of erythrocyte indices, the concentration of the main pro-inflammatory cytokines – interleukin (IL) 1, IL-6. Results. In laboratory rats with NAFLD, a statistically significant increase of intrahepatic enzymes in blood serum was found. The state of the erythrocyte lineage of hematopoiesis in the experimental group progressively worsened and caused the development of anemic syndrome. Synchronously, a statistically significant increase in serum levels of IL-1, IL-6 was recorded, which confirms the correlation of NAFLD with anemia of chronic inflammation.Conclusions. A high concentration of IL-1, IL-6 cytokines in NAFLD inhibits iron absorption in the duodenum, leads to the activation of macrophages, blocking the release of iron processed from aging erythrocytes into plasma. Further study of the mechanisms of anemia in NAFLD provides important therapeutic targets in the treatment of both NAFLD and its comorbidities.

Interleukin-6 Stromal Expression is Correlated with Epithelial-Mesenchymal Transition at Tumor Budding in Colorectal Cancer.

Background. Tumor budding is a poor prognostic factor in colorectal adenocarcinoma, but the underlying mechanism remains unclear. Interleukin-6 (IL6) is one of the main cytokines produced by cancer-associated fibroblasts. IL6 is linked with cancer progression and poor prognosis by activating cancer cells and modifying the cancer microenvironment. However, little is known about the expression of IL6 in tumor budding and its association with tumor budding in colorectal adenocarcinoma. Methods. The clinicopathological and prognostic significance of IL6 in tumor budding was examined using a tissue microarray consisting of 36 patient samples of tumor budding in colorectal adenocarcinoma. IL6 mRNA was detected by RNAscope. Patients were stratified into negative and positive IL6 expression groups. Results. IL6 expression was overwhelmingly observed in cancer stroma but was negligible in cancer cells. Tumor budding grade was higher in the IL6-positive group in cancer stroma than in the IL6-negative group (P = .0161), while the IL6-positive group significantly exhibited the epithelial-mesenchymal transition phenotype compared with the IL6-negative group in cancer stroma (P = .0301). There was no significant difference in overall survival between colorectal adenocarcinoma patients in the IL6-positive and -negative groups in cancer stroma. Conclusion. Tumor budding may be affected by IL6 expression, and IL6 expression in cancer stroma at tumor budding may be an important prognostic marker.

Influence of microbial polyamines on the IL-10 production by peripheral blood leukocytes of healthy donors

As is known, bacterial polyamines, which include cadaverine and putrescine, are capable of influencing the activity of immunocompetent cells in many ways. In particular, this situation is observed in long-term inflammatory diseases, especially with intensive reproduction of microorganisms capable of producing polyamines. It is of interest to study the production of one of the main anti-inflammatory cytokines, IL-10, under the influence of bacterial polyamines. For research, a population of mononuclear leukocytes was isolated from the peripheral blood of healthy donors by gradient centrifugation. The cell suspension was placed in a round-bottomed plates with preliminarily added polyamines at concentrations of 5, 25, 50, 75, and 100 mmol/L. Wells not containing polyamines were used as a control. After incubation for 72 h at 37 °C and 5% CO2, the supernatants were harvested and used to determine the concentration of IL-10. We used a kit for determining the concentration of IL-10 using the enzyme immunoassay method (Russia). Statistical analysis was performed using the Statistica 6.0 software package. In the case of a distribution close to normal, Student’s t-test was used; in the rest, the Mann–Whitney test was used to assess the significance of differences. Studies have shown that leukocytes in the presence of concanavalin A produce IL-10 at a concentration of 17.13±6.08 pg/mL. It has been established that under the influence of polyamines of bacterial origin, the production of IL-10 is enhanced only if putrescine and cadaverine are at concentrations of 50 mmol/L and higher. At low concentrations of polyamines, no significant increase in IL-10 production was detected. Since IL-10 is an anti- inflammatory cytokine, for which the analgesic effect is also known, it should be expected that with an increase in its concentration in the focus of invasion of opportunistic bacteria, the inflammatory process will develop latently, when the symptoms are mild. In general, it can be expected that polyamine-producing bacteria will contribute to the maintenance of few symptomatic inflammation.

Reduced C-reactive protein level at hospital admission in patients treated with Tocilizumab – An attention may be required

BackgroundC-reactive protein (CRP) is a marker of inflammation and infection. The main proinflammatory cytokine that leads to CRP gene expression is IL-6. The study aimed to compare CRP level between patients who were treated with Tocilizumab (TCZ), an il-6 receptor blocker, and other advanced anti-inflammatory treatments (AAIT), as well as with other admitted and non-admitted populations. MethodsA cross-sectional study of all patients (≥18 years) hospitalized at tertiary medical center between December 2009 and February 2020 and treated before hospitalization with (AAIT). Only the first hospitalization of each patient was included. Women admitted to obstetrics department were excluded. Demographic data, first blood tests results, and comorbidities were collected. ResultsThe study included 563 patients treated with AAIT (2.5% received TCZ). Patients treated with TCZ were older (median 75 vs. 50 years, p < 0.001), had higher Charlson score (median 5 vs. 1, p < 0.001) and more infectious diseases at admission (50% vs. 23.4%, p = 0.05). Patients treated with TCZ had lower CRP levels (median 0.5 vs. 25 mg/l, p < 0.001) and more common normal values (64.3% vs. 20.8%, p < 0.001) compared to patients treated with other AAIT.CRP level in patients treated TCZ (median 0.5 mg/l) was lower than that of 58,548 patients admitted to the hospital between 2010 and 2020 (median 12.55 mg/l, p < 0.001) and not statistically different from 140 non-admitted randomly selected individuals without acute disease (1.33 mg/l, p = 0.294). ConclusionTocilizumab is associated with lower levels of CRP in patients admitted to acute care hospital. This finding must be considered by treating physician to avoid misinterpretation of CRP results.

Study of tumor necrosis factor receptor in the inflammatory bowel disease.

Ulcerative colitis (UC) and Crohn's disease (CD) are part of Inflammatory Bowel Diseases (IBD) and have pathophysiological processes such as bowel necrosis and enteric neurons and enteric glial cells. In addition, the main inflammatory mediator is related to the tumor necrosis factor-alpha (TNF-α). TNF-α is a me-diator of the intestinal inflammatory processes, thus being one of the main cytokines involved in the pathogenesis of IBD, however, its levels, when measured, are present in the serum of patients with IBD. In addition, TNF-α plays an important role in promoting inflammation, such as the production of interleukins (IL), for instance IL-1β and IL-6. There are two receptors for TNF as following: The tumor necrosis factor 1 receptor (TNFR1); and the tumor necrosis factor 2 receptor (TNFR2). They are involved in the pathogenesis of IBD and their receptors have been detected in IBD and their expression is correlated with disease activity. The soluble TNF form binds to the TNFR1 receptor with, and its activation results in a signaling cascade effects such as apoptosis, cell proliferation and cytokine secretion. In contrast, the transmembrane TNF form can bind both to TNFR1 and TNFR2. Recent studies have suggested that TNF-α is one of the main pro-inflammatory cytokines involved in the pathogenesis of IBD, since TNF levels are present in the serum of both patients with UC and CD. Intravenous and subcutaneous biologics targeting TNF-α have revolutionized the treatment of IBD, thus becoming the best available agents to induce and maintain IBD remission. The application of antibodies aimed at neutralizing TNF-α in patients with IBD that induce a satisfactory clinical response in up to 60% of patients, and also induced long-term maintenance of disease remission in most patients. It has been suggested that anti-TNF-α agents inactivate the pro-inflammatory cytokine TNF-α by direct neutralization, i.e., resulting in suppression of inflammation. However, anti-TNF-α antibodies perform more complex functions than a simple blockade.

Effect of IL6 on the cancer-associated cachexia: a mini review

More than 20% of tumor patients die because of cancer-associated cachexia (CAC) which is considered a multifactorial syndrome that is characterized by loss of weight, adipose tissue, and skeletal muscle, as well as anemia, accompanied by systemic inflammation. CAC is associated with various agents including cytokines. IL-6 is one of the major proinflammatory and pleiotropic cytokines produced by several cell types which induces and blocks diverse signaling pathways related to cancers and their disorders. For instance, tumor-secreted IL6 is the main cytokine that dilapidated the adipocyte in CAC. IL-6 also plays an essential role in skeletal muscle loss. The involvement of IL-6 in cancer progression and metastasis emerges the hypothesis of its role as a prognostic factor or a treatment agent in CAC. So, with regard to the importance of CAC and its high incidence, it seems that a comprehensive study on the role and mechanism of action of IL-6 in CAC can fill the scientific gap in death resulting from CAC.

Prognostic immune markers identifying patients with severe COVID-19 who respond to tocilizumab.

A growing number of evidences suggest that the combination of hyperinflammation, dysregulated T and B cell response and cytokine storm play a major role in the immunopathogenesis of severe COVID-19. IL-6 is one of the main pro-inflammatory cytokines and its levels are increased during SARS-CoV-2 infection. Several observational and randomized studies demonstrated that tocilizumab, an IL-6R blocker, improves survival in critically ill patients both in infectious disease and intensive care units. However, despite transforming the treatment options for COVID-19, IL-6R inhibition is still ineffective in a fraction of patients. In the present study, we investigated the impact of two doses of tocilizumab in patients with severe COVID-19 who responded or not to the treatment by analyzing a panel of cytokines, chemokines and other soluble factors, along with the composition of peripheral immune cells, paying a particular attention to T and B lymphocytes. We observed that, in comparison with non-responders, those who responded to tocilizumab had different levels of several cytokines and different T and B cells proportions before starting therapy. Moreover, in these patients, tocilizumab was further able to modify the landscape of the aforementioned soluble molecules and cellular markers. We found that tocilizumab has pleiotropic effects and that clinical response to this drug remain heterogenous. Our data suggest that it is possible to identify patients who will respond to treatment and that the administration of tocilizumab is able to restore the immune balance through the re-establishment of different cell populations affected by SARS-COV-2 infection, highlighting the importance of temporal examination of the pathological features from the diagnosis.

Role of Fibroblasts in Chronic Inflammatory Signalling in Chronic Rhinosinusitis with Nasal Polyps-A Systematic Review.

Chronic rhinosinusitis with nasal polyps (CRSwNP) is an inflammatory disease of the nose and paranasal sinuses characterized by the presence of nasal polyps. The symptoms produced by the presence of nasal polyps such as nasal obstruction, nasal discharge, facial pain, headache, and loss of smell cause a worsening in the quality of life of patients. The source of the nasal polyps remains unclear, although it seems to be due to a chronic inflammation process in the sinonasal mucosa. Fibroblasts, the main cells in connective tissue, are intimately involved in the inflammation processes of various diseases; to this end, we carried out a systematic review to evaluate their inflammatory role in nasal polyps. Thus, we evaluated the main cytokines produced by nasal polyp-derived fibroblasts (NPDF) to assess their involvement in the production of nasal polyps and their involvement in different inflammatory pathways. The results of the review highlight the inflammatory role of NPDF through the secretion of various cytokines involved in the T1, T2, and T3 inflammatory pathways, as well as the ability of NPDF to be stimulated by a multitude of substances. With these findings, the fibroblast is positioned as a new potential therapeutic target in the treatment of CRSwNP.

Autoimmune Valvular Carditis Requires Endothelial Cell TNFR1 Expression.

Inflammation is a key driver of cardiovascular pathology, and many systemic autoimmune/rheumatic diseases are accompanied by increased cardiac risk. In the K/B.g7 mouse model of coexisting systemic autoantibody-mediated arthritis and valvular carditis, valve inflammation depends on macrophage production of TNF (tumor necrosis factor) and IL-6 (interleukin-6). Here, we sought to determine if other canonical inflammatory pathways participate and to determine whether TNF signaling through TNFR1 (tumor necrosis factor receptor 1) on endothelial cells is required for valvular carditis. We first asked if type 1, 2, or 3 inflammatory cytokine systems (typified by IFNγ, IL-4, and IL-17, respectively) were critical for valvular carditis in K/B.g7 mice, using a combination of in vivo monoclonal antibody blockade and targeted genetic ablation studies. To define the key cellular targets of TNF, we conditionally deleted its main proinflammatory receptor, TNFR1, in endothelial cells. We analyzed how the absence of endothelial cell TNFR1 affected valve inflammation, lymphangiogenesis, and the expression of proinflammatory genes and molecules. We found that typical type 1, 2, and 3 inflammatory cytokine systems were not required for valvular carditis, apart from a known initial requirement of IL-4 for autoantibody production. Despite expression of TNFR1 on a wide variety of cell types in the cardiac valve, deleting TNFR1 specifically on endothelial cells protected K/B.g7 mice from valvular carditis. This protection was accompanied by reduced expression of VCAM-1 (vascular cell adhesion molecule), fewer valve-infiltrating macrophages, reduced pathogenic lymphangiogenesis, and diminished proinflammatory gene expression. TNF and IL-6 are the main cytokines driving valvular carditis in K/B.g7 mice. The interaction of TNF with TNFR1 specifically on endothelial cells promotes cardiovascular pathology in the setting of systemic autoimmune/rheumatic disease, suggesting that therapeutic targeting of the TNF:TNFR1 interaction could be beneficial in this clinical context.

The interweaved signatures of common-gamma-chain cytokines across immunologic lineages.

"γc" cytokines are a family whose receptors share a "common-gamma-chain" signaling moiety, and play central roles in differentiation, homeostasis, and communications of all immunocyte lineages. As a resource to better understand their range and specificity of action, we profiled by RNAseq the immediate-early responses to the main γc cytokines across all immunocyte lineages. The results reveal an unprecedented landscape: broader, with extensive overlap between cytokines (one cytokine doing in one cell what another does elsewhere) and essentially no effects unique to any one cytokine. Responses include a major downregulation component and a broad Myc-controlled resetting of biosynthetic and metabolic pathways. Various mechanisms appear involved: fast transcriptional activation, chromatin remodeling, and mRNA destabilization. Other surprises were uncovered: IL2 effects in mast cells, shifts between follicular and marginal zone B cells, paradoxical and cell-specific cross-talk between interferon and γc signatures, or an NKT-like program induced by IL21 in CD8+ T cells.

Effect of transduced mesenchymal stem cells with IL-10 gene on control of allergic asthma.

Asthma is an important pulmonary disease associated with T helper lymphocyte (Th)2 dominant immune response, which can initiate allergic and inflammatory reactions. Interleukin (IL)-10 is the main immune suppressor cytokine, and mesenchymal stem cells (MSCs) have an immune-modulatory potential that can be transduced with the expression of the IL-10 gene to control pathophysiology of allergic asthma. Bone marrow's MSCs were isolated and transduced with the expression vector that contains the expressible IL-10 gene. Then, allergic asthma mouse model was produced and treated with manipulated MSCs. Methacholine challenge test; measurement of IL-4, IL-5, IL-8, IL-13, IL-25, and IL-33; and total and ovalbumin (OVA)-specific immunoglobulin (Ig)E levels were done. Hyperplasia of the goblet cell, secretion of mucus, and peribronchiolar and perivascular eosinophilic inflammation were evaluated in lung pathological sections. IL-25, IL-33, and total IgE levels; AHR; eosinophilic inflammation; hyperplasia of the goblet cell; and secretion of mucus could be controlled in M, MV, and MV-10 groups, and the control in the MV-10 group was strong compared to M and MV groups. MSCs have immune-modulatory capacity that can control allergic asthma pathophysiology, and this effect can be strengthened and reinforced by the expression of IL-10 gene.

THE ROLE OF PROINFLAMMATORY CYTOKINES INTERLEUKIN 1-Β AND TUMOR NECROSIS FACTOR-Α IN DIAGNOSTICS OF PUBERTAL MENORRHAGES AGAINST THYROID PATHOLOGY

Introduction. During puberty, the reproductive system is vulnerable to the influence of any adverse factors that lead to a disorder of its functional state and to menstrual disorders in particular. Among the numerous factors that provoke menstrual dysfunction is the pathology of the thyroid gland.Cytokines are known to be involved in all aspects of innate and acquired immunity, including the activation of growth and differentiation of immunocompetent cells, inflammation and restoration of the function of the affected organ. Cytokines are characterized by the action of preventing the interaction of cells of the immune, hematopoietic, endocrine and nervous systems. Cytokines of the first generation are conditionally distinguished, which are produced by cells of nonspecific anti-infective protection. The main pro-inflammatory cytokines are IL-1β, IL-2, IL-6, interferons, TNF-α and others. The main factor of inflammatory reactions is multifunctional IL-1β. Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine, one of the central regulators of factors and mechanisms of innate resistance. It has many biogenic effects, most of which are similar to the action of IL-1β.Considering the above exactly IL-1β and TNF-α to study their concentration in the blood of adolescent girls with pubertal menorrhagia has been the basis of immunological studies.The aim of the study. Assessment of cytokine status in girls of pubertal age against the background of concomitant thyroid pathology.Material and methods. We examined 70 adolescent girls with pubertal menorrhagia who were treated in the gynecological department of the Chernivtsi Regional Perinatal Center. The girls were divided into two groups: I (main) included 30 adolescent girls diagnosed with pubertal menorrhagia on the background of concomitant thyroid pathology, II group (comparison) - 40 adolescent girls diagnosed with pubertal menorrhagia. Control group III - 25 practically healthy adolescent girls. Pro-inflammatory cytokines, namely interleukin 1-beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) were studied once, after inclusion of patients in the study, by enzyme-linked immunosorbent assay based on a solid-phase ‘sandwich’ variant using mono- and polyclonal antibodies to IL-1β and TNF-α. The tubes with the serum samples were closed with lids and stored in the freezer until analysis at -20 °C.Statistical processing of the material was carried out using the computer program STATISTICA and Microsoft Exel Windows, StatSoft® Inc.The design of the study and all methods used in this study were reviewed and approved by the Bioethics Committee of the Higher Educational Institution “Bukovinian State Medical University” (protocol No. 1, dated 24.01.2011). Research work “Prevention, diagnostics and treatment of perinatal and reproductive system disorders of women and adolescent girls” (№ 201110Н state registration number 0111U006499. Term of realization: 02.2011-12.2015.)Results of the study. The obtained results of the study of the cytokine cascade showed that upon admission to inpatient treatment and examination in the blood of adolescent girls with pubertal menorrhagia without concomitant pathology, the concentration of IL-β increases significantly (by 60.61%) and a tendency to increase (4.09 times) the concentration of TNF-α in the peripheral blood of the examined patients is formed. However, the results of clinical and laboratory examinations obtained during admission for inpatient examination and treatment of adolescent girls with pubertal menorrhagia associated with thyroid pathology showed that patients have a steady tendency to decrease the concentration of IL-1β by 6.96% and TNF-α - by 1.48 times. It was found that the pathology of the thyroid gland contributes to the inhibition of immunocompetent cells production of IL-1β by 1.66 times, TNF-α by 6.04 times.Conclusion. Pathology of the thyroid gland leads to a decrease in immunoregulatory function due to a decrease in the concentration of some important proinflammatory cytokines (IL-1β and TNF-α).

Blocking the action of NLRP3 inflammasome by inhaled ibuprofenate on pulmonary macrophages infected by SARS‐CoV‐2

We present a Graphical Abstract that would represent the potential therapeutic effects of inhaled ibuprofenate on lung macrophages infected by SARS-CoV-2 and its link with the NLRP3 inflammasome. Furthermore, in the manuscript we will provide data on observational studies in pneumonias and Cards. In this manuscript, we will describe the possible mechanisms of action of the non-steroidal anti-inflammatory drug (NSAID) inhaled sodium ibuprofenate in hypertonic saline formulation (NaIHS), through its anti-inflammatory and immunomodulatory therapeutic effects, focusing primarily on the potential blockade of NOD-like receptor protein 3 (NLRP3) inflammasome action in Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2)-infected lung macrophages. Currently, there are available three observational studies in the compassionate use modality in moderate and severe Coronavirus Disease 2019 (COVID-19) pneumonia, carried out in Argentina,1-3 with all three studies concluding that there was a significant reduction in mortality.1-3 The most recent of these was conducted with the approval of the Institutional Review Board of the National Bureau of Economic Research (NBER) of Harvard and Columbia Universities, which analysed data from 5146 patients and concluded that NaIHS reduced mortality by 48.7%,3 although randomized clinical trials are steel needed to confirm these data. NLRP3 inflammasome is a cytosolic multiprotein complex that plays a key role in the regulation of the innate immune system and inflammatory signalling, mostly against viral infections and cell damage.4 Their activation and assembly occurs mainly through different insults: activation of pattern recognition receptors (PRRs) through the pathogen-associated molecular patterns (PAMPs) and the danger-associated molecular patterns (DAMPs), both at the plasma membrane by toll-like receptor 4 (TLR4), as well as at the cytosolic level by nod-like receptor 2 (NLR2), hypoxaemia, renin-angiotensin system (RAS) imbalance, mitochondrial dysfunction and oxidative stress, lysosomal leakage with cathepsin-B release, ion fluxes and others.4 In the face of infection of lung macrophages by SARS-CoV-2, hyperactivation and assembly of the NLRP3 inflammasome is triggered, resulting in a cascade of positive caspase-1 signalling (first step), pro-inflammatory cytokines and inflammatory signalling pathways.4 This way, the activated NLRP3 inflammasome leads to the maturation and release by proteolytic cleavage of immature precursors of: caspase-1, interleukin-1 beta (IL-1β), IL-18 and gasdermin-D (GSDMD).4 The NLRP3 inflammasome promotes, on one arm, the release and activation of IL-1β and IL-18 and these cytokines, in turn, lead to the activation of the nuclear factor kappa light chain enhancer of activated B cells (NF-κB), the stimulation of natural killer (NK) cells, the release of interferon-γ (IFN-γ) and the inhibition of IL-33 activity.4 In addition, on the other arm, the NLRP3 inflammasome promotes GSDMD-mediated release and activation of IL-1β and IL-18 through membrane pores and it also leads to the release and activation of GSDMD-NT, which together with caspase-4 and caspase-5 would lead to T-cell pyroptosis.4 This cascade of events triggers the activation and recruitment of pulmonary macrophages, generating the cytokine storm, activation of inflammatory signalling pathways and also provoking a positive immune-inflammatory amplifying feed-back loop between all the above-mentioned components.4 It might block NLRP3 inflammasome action through the following effects: In vitro and in vivo, ibuprofen inhibits the activation of NLRP3 inflammasome6 and thus significantly reduces the levels of caspase-1,6 IL-1β6 and IL-18.6 It might inhibit NLRP3 inflammasome due to its peroxisome proliferator-activated receptor-gamma (PPAR-γ) agonist effect,4, 7 and it also might inhibit NLRP3 inflammasome via inhibition of membrane volume anion chloride channel and thus chloride efflux.4, 8 It might prevent pyroptosis through negative modulation of the inflammatory signalling pathway phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR),4, 7 caspase-14, 6, caspase-44, 9 and caspase-54, 9. It might negatively modulate the activation and recruitment of pulmonary macrophages.6 It would negatively modulate the main inflammatory signalling pathways: COX25, NF-κB,5 signal transducer and activator of transcription 3 (STAT3),5 TLR45 and PI3K/AKT/mTOR7. It would negatively modulate the main pro-inflammatory cytokines: IL-1β,5 tumor necrosis factor-alpha (TNF-α),5 IL-65, IFN-γ,5 transforming growth factor-beta 1 (TGF-β1),5 intercellular adhesion molecule-1 (ICAM-1)5 and IL-186. It may revert hypoxaemia, exclusively of the vasoplegic type10 and thus, it might prevent NLRP3 inflammasome activation. It would restore the RAS imbalance5 and thus, it also might prevent NLRP3 inflammasome activation. We have herein postulated the potential therapeutic effects of adjuvant treatment with NaIHS that might block NLRP3 inflammasome action in SARS-CoV-2-infected lung macrophages, and it might negatively modulate activation with monocyte recruitment, cytokine storm and activation of inflammatory signalling pathways. And above all, NaIHS has demonstrated in the aforementioned observational studies: a significant reduction in mortality, improvement of clinical outcomes, rapid reversal of hypoxaemia and decrease in hospitalization stay in pneumonia and Coronavirus Disease 2019-associated Acute Respiratory Distress Syndrome (CARDS). Yet, randomised clinical trials are still needed to confirm these emerging data. The authors declare that they have no conflicts of interest to this manuscript. The authors received no funding for this article.

Different peripheral expression patterns of the nicotinic acetylcholine receptor in dementia with Lewy bodies and Alzheimer’s disease

BackgroundThe diffuse distribution of nicotinic cholinergic receptors (nAChRs) in both brain and peripheral immune cells points out their involvement in several pathological conditions. Indeed, the deregulated function of the nAChR was previously correlated with cognitive decline and neuropsychiatric symptoms in Alzheimer’s disease (AD) and Dementia with Lewy bodies (DLB).The evaluation in peripheral immune cells of nAChR subtypes, which could reflect their expression in brain regions, is a prominent investigation area.ObjectivesThis study aims to evaluate the expression levels of both the nAChR subunits and the main known inflammatory cytokines in peripheral blood mononuclear cells (PBMCs) of patients with DLB and AD to better characterize their involvement in these two diseases.ResultsHigher gene expression levels of TNFα, IL6 and IL1β were observed in DLB and AD patients in comparison with healthy controls (HC). In our cohort, a reduction of nAChRα4, nAChRβ2 and nAChRβ4 was detected in both DLB and AD with respect to HC. Considering nAChR gene expressions in DLB and AD, significant differences were observed for nAChRα3, nAChRα4, nAChRβ2 and nAChRβ4 between the two groups. Moreover, the acetylcholine esterase (AChE) gene expression was significantly higher in DLB than in AD. Correlation analysis points out the relation between different nAChR subtype expressions in DLB (nAChRβ2 vs nAChRα3; nAChRα4 vs nAChRα3) and AD (nAChRα4 vs nAChRα3; nAChRα4 vs nAChRβ4; nAChRα7 vs nAChRα3; nAChRα7 vs nAChRα4).ConclusionsDifferent gene expressions of both pro-inflammatory cytokines and nAChR subtypes may represent a peripheral link between inflammation and neurodegeneration. Inflammatory cytokines and different nAChRs should be valid and accurate peripheral markers for the clinical diagnosis of DLB and AD. However, although nAChRs show a great biological role in the regulation of inflammation, no significant correlation was detected between nAChR subtypes and the examined cytokines in our cohort of patients.

Koena, a Novel Compound That Dampens Inflammation in the Skin

Inflammatory skin diseases are characterized by the activation of the innate and adaptive immune system via the production of pro-inflammatory cytokines. The main proinflammatory cytokines responsible for this development include TNF-alpha, Interleukin 1, 6 and 17, also the transcription factor NF-kappa B. Small molecule anti-inflammatory compounds such as those detailed in the paper, show much promise in dealing with these skin disorders. Four nutraceutical molecules that are well characterized by their capacity to interact with many of the pro-inflammatory cytokines, importantly IL-17, are found in an Australian product, Koena. Koena has been shown to not only decrease these cytokines but also help increase the production of Interleukin 10 and important anti-inflammatory cytokines. This paper reports clinical benefits of Koena in a variety of circumstances; eczema and psoriasis; insect bites and sting; solar damaged skin and keratosis. An analysis of patient/purchaser reviews via the website Koena.com.au to ascertain what condition they purchased the product for, if any, and the satisfaction rating after use. More than 35% of reviewers reported purchasing Koena to help with inflamed skin. Those reporting being satisfied with the result was 95% for this use. This paper reports on the benefit of small molecule anti-inflammatories in modulating and not blocking an immune response. It also illustrates the benefits of multitherapy when dealing with complex inflammatory responses as opposed to monotherapy.

Expression of Circulatory Interleukin-6 Concentration Associated with Pseudomonas aeruginosa Persistence in Recurrent Urinary Tract Infections

Background: The main superfamily cytokine member is interleukin-6 (IL-6). IL-6, promptly produced in response to infections and tissue injuries, contributes to host defense through the stimulation of acute phase responses, hematopoiesis, and immune reactions. Objectives: To investigate the role of IL-6 as a biomarker in recurrent urinary tract infections (RUTIs). The real-time polymerase chain reactions were performed by using specific primers with reference gene GAPDH and the target genes IL-6. Materials and Methods: A case–control study was done in Babylon city hospital from February 2021 to March 2022. A total of 110 patients with RUTIs with different age and sex, and healthy individuals as control group were enrolled in this study. Midstream urine was taken for culturing and identification of Pseudomonas aeruginosa. Blood samples were obtained from all the patients and the secreted IL-6 levels were detected by enzyme linked immunosorbent assay (ELISA). RNA was extracted for gene expression. Results: The expression of IL-6 was increased more than % 30-fold in UTIs compared with control group. Also, the results found that serum concentrations of IL-6 detected by ELISA assay showed significant differences at P &lt; 0.001 for the patients compared with control group. Conclusions: Interleukin-6 gene expression shows up regulation in RUTI caused by P. aeruginosa.

Pancreatic Islet Cells Response to IFNγ Relies on Their Spatial Location within an Islet

Type 1 diabetes (T1D) is an auto-immune disease characterized by the progressive destruction of insulin-producing pancreatic beta cells. While beta cells are the target of the immune attack, the other islet endocrine cells, namely the alpha and delta cells, can also be affected by the inflammatory milieu. Here, using a flow cytometry-based strategy, we compared the impact of IFNγ, one of the main cytokines involved in T1D, on the three endocrine cell subsets isolated from C57BL/6 mouse islets. RNA-seq analyses revealed that alpha and delta cells exposed in vitro to IFNγ display a transcriptomic profile very similar to that of beta cells, with an increased expression of inflammation key genes such as MHC class I molecules, the CXCL10 chemokine and the programmed death-ligand 1 (PD-L1), three hallmarks of IFNγ signaling. Interestingly, at low IFNγ concentration, we observed two beta cell populations (responders and non-responders) based on PD-L1 protein expression. Our data indicate that this differential sensitivity relies on the location of the cells within the islet rather than on the existence of two different beta cells subsets. The same findings were corroborated by the in vivo analysis of pancreatic islets from the non-obese diabetic mouse model of T1D, showing more intense PD-L1 staining on endocrine cells close to immune infiltrate. Collectively, our work demonstrates that alpha and delta cells are as sensitive as beta cells to IFNγ, and suggests a gradual diffusion of the cytokine into an islet. These observations provide novel insights into the in situ inflammatory processes occurring in T1D progression.

Impacts of stress-induced inflammation on feed intake of beef cattle

Livestock animals are often exposed to unavoidable stressful situations during their productive life that triggers stress-induced inflammatory responses, which are known to influence their nutrient requirements and feed intake. Decreased growth performance and immunocompetence of stressed livestock are often the main consequence of reduced feed intake. Because feed intake is usually reduced in animals experiencing stress conditions, concentrations of certain nutrients in the diets typically need to be increased to meet the requirements of the animals. Therefore, understanding the mechanisms that control feed intake in animals experiencing stress-induced inflammation is essential for increasing intake, milk or meat production, feed efficiency, and animal health. This review highlights the hormones regulating feed intake in ruminants and how stress-induced inflammation affect these hormones at local and systemic levels. The mechanism of feed intake regulation in ruminants is extremely complex and involves multiple controls. The liver is an important sensor of energy status in animals under homeostatic conditions, which transmits signals to brain feeding centers that modulate appetite. However, the physiologic consequences associated with different stressors will rearrange the hierarchy of mechanisms controlling feed intake compared to animals under homeostatic conditions, and other tissues (e.g., intestines), systems (e.g., endocrine and lymphatic) hormones (e.g., leptin and ghrelin) will directly affect intake regulation during stress and inflammatory conditions. It is suggested that the immune system can interact with the central nervous system to modulate feed intake. As example, stress events elicit numerous stressors that increase circulating proinflammatory cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-8, and acute-phase proteins (APP), and the magnitude of these responses are negatively correlated with feed intake. A direct effect of these cytokines on rumen microbial fermentation and intestinal barrier function was also reported and might indirectly affect intake regulation in ruminants. This review describes the main hormones and proinflammatory cytokines involved in stress-induced inflammation and how they can directly or indirectly affect intake regulation in ruminants. Understanding the mechanisms controlling feed intake in ruminants will help producers to implement management and feed strategies to optimize productivity and profitability in stressed livestock species.