Cytokine Synthesis Research Articles

Bupropion Associated Immunomodulatory Effects on Peripheral Cytokines in Male with Major Depressive Disorder

Background: Experimental and clinical studies have reported increased levels of pro-inflammatory cytokines in patients with major depressive disorder (MDD), suggesting that immune system dysregulation may contribute to MDD pathophysiology. Aim: Due to the lack of knowledge about the immune potential of antidepressants, this study investigated the immunomodulatory effects of bupropion, a norepinephrine–dopamine reuptake inhibitor. Methods: This study involved 18 patients with MDD treated with bupropion (150 mg/d) for 4 weeks and 23 healthy volunteers. All participants underwent multiplex bead-based cytokine assessment before and after bupropion treatment to quantify the following cytokines: interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, interferon-γ, tumor necrosis factor-α, granulocyte colony-stimulating factor, granulocyte-macrophage CSF, monocyte chemotactic protein-1, and macrophage inflammatory protein-1β. Results: Four-week treatment with bupropion significantly increased the levels of IL-1β (P = 0.011), IL-4 (P = 0.019), IL-5 (P = 0.019), IL-7 (P = 0.021), and IL-8 (P = 0.023) compared to the control group. Furthermore, the percentage change in most cytokines, including anti-inflammatory cytokines such as IL-4, IL-5, IL-10, and IL-13, was significantly increased after bupropion treatment. Conclusion: The promoted synthesis of anti-inflammatory cytokines to surpass the pro-inflammatory cytokines may be a crucial step in the treatment of MDD patients with bupropion.

PSIX-9 Activation of Multiple Inflammatory Signaling Pathways May Contribute to the Pathogenesis of Idiopathic Cystitis in Felines

Abstract Feline idiopathic cystitis (FIC) is a common urinary disorder in domestic cats. Clinical signs include straining or failing to urinate, frequent urination, and discolored urine. Inflammation is a key characteristic in the pathogenesis of cystitis. We assessed circulating inflammatory markers in a retrospective study in cats housed in the Hill’s colony diagnosed with cystitis (n = 4; 15.2 to 18.6yr) and in control cats (n = 8; 5.9 to 16.5 yr). Blood samples were collected at end-of-life immediately before necropsy after they had lived their full lives. End-of-life pathology reports indicated inflammation and presence of inflammatory infiltrates in the bladder with some cats also showing renal inflammation. RNA was extracted from blood collected in PAXgene tubes, and gene expression investigated using NanoString nCounter platform. Analysis was performed using the nSolver software. There was a significant increase in IRAK4, a kinase that activates the NFκB pro-inflammatory signaling pathway, in FIC when compared with controls (1.56, P < 0.05). Small molecule inhibitors of IRAK4 are being developed as targets of inflammation. Signaling molecules in the NFκB pathway were also significantly increased including Rel A (p65 subunit; 1.3 fold), NFκB1 (p100 subunit; 1.3 fold), IKBKB (1.29 fold), and TANK (1.46 fold) in FIC cats (all P < 0.05). Another signaling pathway contributing to pro-inflammatory signaling is the JAK/STAT pathway. There was a significant increase in JAK2 (1.65) and STAT3 (1.48). Blockade of JAK2/STAT3 pathway can attenuate cytokine synthesis and reduce inflammation. STAT6 was also significantly increased in FIC cats (1.35). Activation of STAT6, by IL-4 and IL-13, can lead to systemic inflammation. Activation of the complement immune pathway was also seen in FIC. While the complement system is important for an innate immune response, its sustained activation leads to chronic pathological inflammation. There was a significant increase in complement C1s, an initiator of classical complement activation pathway, in FIC cats (1.43; P < 0.05). A late component, complement C7, was also upregulated in FIC (2.01 fold; P < 0.05). C7 binds to the C5b-6 complex which eventually becomes part of the terminal complement complex. Increased abundance of C7 and eventual terminal complement complex are associated with increased inflammation and may be important in the progression of FIC. There was also an increase in PECAM1 in FIC cats (1.87; P < 0.05). Signaling through PECAM1 leads to the activation of neutrophils and monocytes. DUSP1 was also elevated in FIC (2.24; P < 0.05). Increased abundance of DUSP1 is hypothesized to be a predictor of chronic subclinical inflammation in humans with cardiovascular disease. Our results identify activation of pro-inflammatory signaling pathways that may be potential targets for alleviating inflammation in cystitis. Pathogenesis of FIC is likely multi-factorial, and nutritional intervention to attenuate the activation of inflammatory pathways may be an important strategy in the management of cystitis.

In silico molecular docking studies of 2, 4 and 2, 6-di-tert-butylphenol against NAGK and SPP1 proteins to recuperate rheumatoid arthritis

Rheumatoid arthritis (RA) is a chronic autoimmune disorder that primarily affects the synovial joints. It is connected to progressive disability, early death and socioeconomic burdens. So, there is an urgent need to develop novel drugs that effectively treat patients at each stage of disease progression. N-acetylglucosamine kinase (NAGK) and secreted phosphoprotein 1(SPP1) are two attractive drug targets for RA. NAGK protein converts endogenous N-acetyl-D-glucosamine (GlcNAc) into GlcNAc 6-phosphate. GlcNAc has suppressive effects on experimental RA in mouse models. SPP1 stimulates the synthesis of a key proinflammatory cytokine implicated in the pathogenesis of RA. In order to design new drug candidates, crystal structure of human NAGK (PDB Id: 2CH5) was retrieved from the protein data bank. Amino acid sequence of SPP1 (Uniprot Id: P10451) was retrieved from uniprot database and modeled by the Phyre2 server. Computed SPP1 model energy was minimized using the YASARA energy minimization server and validated by the Ramachandran plot and the ProSA-web error-detection tool. 2, 4-Di-tert-butylphenol and 2, 6-Di-Tert-butylphenol are known compound isolates from bark extract of Schleichera oleosa having anti-arthritic and anti-inflammatory activities as accessed by PASS online server. These compounds were docked against active site of NAGK and SPP1 proteins using MTiAutoDock server. The binding affinity of these compounds against NAGK and SPP1 proteins ranges from -6.12 to -7.17 kcal/mol. The findings of this study will open the way for the development of herbal remedies for rheumatoid arthritis based on the Schleichera oleosa plant, potentially leading to the development of novel drugs.

Cytokine Profile Following Varnimcabtagene Autoleucel (IMN-003A) in Patients with Relapsed Refractory B Cell Malignancies in the First-in-India Industry Phase-2 Study (IMAGINE)

Background: Varnimcabtagene autoleucel (Var-cel, IMN-003A) is an autologous CD19 directed CAR-T cell product with a 4-1BB co-stimulatory domain and a non-FMC63 murine single chain variable fragment (A3B1 binder), manufactured in India, tested in the IMAGINE study (CTRI/2022/03/041162), a phase-2 clinical trial for patients (pts) with relapsed and/or refractory B cell malignancies. This abstract evaluates the cytokine profile and its correlation with safety outcomes and CAR-T expansion following var-cel infusion. Methods: Serum was collected pre-infusion and upto 13 timepoints post infusion as per study schedule for evaluating cytokine levels. Nine cytokines (IFN-g, TNF-α, IL-1β, IL-2, IL-7, IL-8, IL-10, IL-15 and MCP-1) were measured using a multiplexed kit on the Magpix platform. IL-6 level was measured using electrochemiluminescence immunoassay (ECLIA). Safety and adverse event data were collected for analysis along with measuring various cytokine levels in the serum. CAR-T persistence post infusion was monitored using ddPCR. Results: 24 patients (pts) were infused with var-cel in the IMAGINE study. Var-cel levels in peripheral blood post-infusion were measured at multiple timepoints for all infused pts using ddPCR platform. The var-cel levels peaked (Tmax) at a median of 10 days post infusion (D+10; range: 10 - 28 days) with median Cmax of 125,242 (range 18,256 - 413,968 copies/µg genomic DNA). The median duration of persistence was 56 days with persistent CAR seen in 3 pts at last follow-up. Adverse events of special interest (AESIs) reported were CRS (Grade [G] 1 62.5%; G3+ 4.2%; overall 66.7%) and ICANS (G1 4.2%; G3+ 0%; overall 4.2%). The median onset of CRS at D+5, 5 days before the median var-cel Tmax of D+10. The median duration for CRS was 3 days. No G3+ ICANS was reported. Serum IL-6 levels peaked at a median of D+7 post infusion and reached above 40 pg/mL for 13 pts, all of whom experienced CRS. All eight pts with no CRS had peak IL-6 levels below 40 pg/mL. Despite this significant association with CRS (p<0.001, Man-Whitney test) and published causation, IL-6 level peaked towards the end of clinical course of CRS manifestation, but usually at or before peak var-cel expansion (Tmax). Interestingly, the increase in IL-6 levels tended to begin early and persist longer for some B-NHL patients compared to B-ALL patients. Preliminary multiplexed cytokine analysis for evaluated pts (n=6) indicated that IFN-γ, TNF-α, IL-1β, IL-2, IL-7, IL-8 remained within published physiological limits for most patients at all timepoints (except one pt with highest levels at screening). One patient showed mild increase in IFN-γ (peak 50.88 pg/mL) and two pts showed mild increase in IL-8 around D+4 to D+7 (peak 47.71 pg/mL). IL-15 and MCP-1 levels were relatively higher (>2 fold) from D0 to D+7 post infusion, compared to the screening timepoint, consistent with the duration of CRS. Only IL-10, an anti-inflammatory and cytokine synthesis inhibitory factor stood out having a sharp peak (median: 230 pg/mL) for all six patients at D+4 to D+10, consistent with the timeline for end of CRS and before var-cel Tmax. Occasional pt with no clinical CRS also showed IL-10 peak. Updated cytokine profile data shall be presented at the meeting. Conclusions: This is the first-in-India industry study evaluating cytokine profile at multiple timepoints before and after var-cel infusion and its correlation with clinical CRS. As per published literature, IL-6 levels were significantly associated with clinical CRS and showed association with peak var-cel expansion. There was a sharp increase in both IL-6 and IL-10 levels in serum post infusion, concurrent with clinical CRS prior to peak var-cel expansion. A further detailed study of cytokine profile following var-cel infusion shall help prognosticate safety outcomes.

The effect of vitamin D on adolescents' primary dysmenorrhea.

Vitamin D receptor (VDR) expression in the female reproductive tract explains the regulatory role of vitamin D on inflammatory cytokine and prostaglandin (PGD) synthesis. This study aimed to evaluate the effect of vitamin D on adolescents' primary dysmenorrhea and the relationship between Vit. D and adolescents' primary dysmenorrhea. Eighty-five adolescents were included in the current study. After a detailed evaluation, pelvic sonography was performed for all participants to rule out any pelvic pathology. Blood samples were collected to measure thyroid stimulating hormone (TSH), prolactin, glycosylated hemoglobin (HbA1C), and 25-hydroxyvitamin D (25[OH]D). Participants were administered vitamin D (50,000 IU weekly for five months), and their dysmenorrhea symptoms were evaluated before and after this period using the Visual Analog Scale (VAS) and the Verbal Multidimensional Scoring (VMS). The mean VAS and VMS scores of dysmenorrhea statistically decreased from 8.7±0.91 and 2.65±0.93 to 4.8±0.75 and 0.80±0.75, respectively, after vitamin D intake (p=0.03 and 0.025, respectively). Significant negative associations between 25(OH)D and VAS (R = -0.886; p<0.00001) and VMS of dysmenorrhea (R = -0.885; p<0.00001) were detected in this study. Vit. D could be a useful therapeutic option to reduce the severity of primary dysmenorrhea and could limit the use of non-steroidal anti-inflammatory drugs.

6-Hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline Demonstrates Anti-Inflammatory Properties and Reduces Oxidative Stress in Acetaminophen-Induced Liver Injury in Rats.

We examined the effects of 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline on markers of liver injury, oxidative status, and the extent of inflammatory and apoptotic processes in rats with acetaminophen-induced liver damage. The administration of acetaminophen caused the accumulation of 8-hydroxy-2-deoxyguanosine and 8-isoprostane in the liver and serum, as well as an increase in biochemiluminescence indicators. Oxidative stress resulted in the activation of pro-inflammatory cytokine and NF-κB factor mRNA synthesis and increased levels of immunoglobulin G, along with higher activities of caspase-3, caspase-8, and caspase-9. The administration of acetaminophen also resulted in the development of oxidative stress, leading to a decrease in the level of reduced glutathione and an imbalance in the function of antioxidant enzymes. This study discovered that 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline reduced oxidative stress by its antioxidant activity, hence reducing the level of pro-inflammatory cytokine and NF-κB mRNA, as well as decreasing the concentration of immunoglobulin G. These changes resulted in a reduction in the activity of caspase-8 and caspase-9, which are involved in the activation of ligand-induced and mitochondrial pathways of apoptosis and inhibited the effector caspase-3. In addition, 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline promoted the normalization of antioxidant system function in animals treated with acetaminophen. As a result, the compound being tested alleviated inflammation and apoptosis by decreasing oxidative stress, which led to improved liver marker indices and ameliorated histopathological alterations.

CD137L Inhibition Ameliorates Hippocampal Neuroinflammation and Behavioral Deficits in a Mouse Model of Sepsis-Associated Encephalopathy

Anxiety manifestations and cognitive dysfunction are common sequelae in patients with sepsis-associated encephalopathy (SAE). Microglia-mediated inflammatory signaling is involved in anxiety, depression, and cognitive dysfunction during acute infection with bacterial lipopolysaccharide (LPS). However, the molecular mechanisms underlying microglia activation and behavioral and cognitive deficits in sepsis have not been in fully elucidated. Based on previous research, we speculated that the CD137 receptor/ligand system modulates microglia function during sepsis to mediate classical neurological SAE symptoms. A murine model of SAE was established by injecting male C57BL/6 mice with LPS, and cultured mouse BV2 microglia were used for in vitro assays. RT-qPCR, immunofluorescence staining, flow cytometry, and ELISA were used to assess microglial activation and the expression of CD137L and inflammation-related cytokines in the mouse hippocampus and in cultured BV2 cells. In addition, behavioral tests were conducted in assess cognitive performance and behavioral distress. Immunofluorescence and RT-qPCR analyses showed that hippocampal expression of CD137L was upregulated in activated microglia following LPS treatment. Pre-treatment with the CD137L neutralizing antibody TKS-1 significantly reduced CD137L levels, attenuated the expression of M1 polarization markers in microglia, and inhibited the production of TNF-α, IL-1β, and IL-6 in both LPS-treated mice and BV2 cells. Conversely, stimulation of CD137L signaling by recombinant CD137-Fc fusion protein activated the synthesis and release of pro-inflammatory cytokines in cultures BV2 microglia. Importantly, open field, elevated plus maze, and Y-maze spontaneous alternation test results indicated that TKS-1 administration alleviated anxiety-like behavior and spatial memory decline in mice with LPS-induced SAE. These findings suggest that CD137L upregulation in activated microglia critically contributes to neuroinflammation, anxiety-like behavior, and cognitive dysfunction in the mouse model of LPS-induced sepsis. Therefore, therapeutic modulation of the CD137L/CD137 signaling pathway may represent an effective way to minimize brain damage and prevent cognitive and emotional deficits associated with SAE.

Bacteria-Targeting Nanosilver-Based Antibacterial Drugs for Efficient Treatment of Drug-Resistant Bacterial-Infected Keratitis.

Keratitis caused by drug-resistant bacteria is a severe condition that can lead to corneal perforation and even blindness, making effective treatment a top priority amid growing antibiotic resistance. Eye drops for anti-inflammatory treatment necessitate frequent administration of high doses throughout every day due to bacterial resistance resulting from antibiotic overuse and the low bioavailability of drugs. To overcome these issues, an antibacterial nanocomposite is prepared via conjugating random copolymers of galactose and 3-(acrylamide)phenylboronic acid to the surface of silver nanoparticles. The customized nanocomposites trigger specific binding to bacteria, resulting in excellent retention of the drug on the ocular surface, resulting in rapid and powerful killing of bacteria and inhibition of bacterial proliferation. Due to its superior drug delivery capabilities to the ocular surface, the functionalized nanocomplex markedly amplifies the anti-inflammatory efficacy, even at low doses. This effect is achieved by impeding immune cell infiltration and diminishing the synthesis of inflammatory mediators and cytokines, thereby suggesting enhanced healing properties for corneal inflammation. This study demonstrates a promising nanocomposite which is an effective and safe antibacterial strategy for bacterial keratitis with favorable prognostic and clinical conversion potential.

Aflatoksin: Aspek kesehatan, metode reduksi dan deteksinya

Food contaminant aflatoxin is a mycotoxin produced by Aspergillus flavus and A. parasiticus. This article aims to discover the pathophysiological, immunological, pharmacological, toxicity, and reduction and detection methods of aflatoxin. Kinds of literature were obtained from Research Gate, Pubmed, and Science Direct with the primary keyword “aflatoxin”. Pathophysiologically, aflatoxin-contaminated food has been proven to cause necrosis in liver cells, developing into liver cancer. Immunologically, aflatoxin decreases monocyte and dendritic cell phagocytosis, neutrophil cell ATP production, and pro-inflammatory cytokine synthesis. Aflatoxin toxicity is at the LD50 of 12-16 mg/kg b.w, causing death. Pharmacologically, 120-201 µg/kg b.w causes aflatoxicosis, in vivo studies indicated that NovaSil, Sulfarophan, and Monanthotaxis caffra leaf extract may reduce its toxicity. Aflatoxins are highly thermostable; hence, once food has been contaminated, they cannot be destroyed by normal cooking process. The control and reduction should be conducted in post-harvest handling, common physical means practiced are heating, drying, and smoking. Chemically using ozone, 0.5 sodium bisulfate, 1% sodium hydroxide, 5% acetic acid, and prochloraz. Biologically using Flavobacterium aurantiacum B-184, Bacillus velezensis DY3108, and, a consortium of Geobacillus and Tepidimicrobium bacteria. Aflatoxin can be detected using TLC, HPLC, MS, ELISA, and UHPLC-ESI MS/MS. The prevention of aflatoxin occurrence is done through good post-harvest handling, good manufacturing practices, and applying regulations accordingly to ensure food products and feed are at acceptable levels of aflatoxin.

IL10 Gene and Neurodegenerative Sclerosis: A Systematic Review and Meta-Analysis

Amyotrophic Lateral Sclerosis (ALS) and Multiple Sclerosis (MS) are incurable degenerative scleroses with unclear etiology. Neuroinflammation is an important factor in the neurodegeneration characteristic of these diseases. Additionally, Interleukin 10 (IL10) can inhibit the synthesis of inflammatory cytokines and plays a protective role against neurodegeneration associated with neuroinflammation. Thus, we developed a systematic review and meta-analysis in order to clarify the relationship between polymorphisms in the IL10 gene and MS and/or ALS. We searched for observational studies in four international databases without time restrictions. Seventeen studies were added to the systematic review and six polymorphisms were observed: IL10-592 (rs1800872; C&gt;A), IL10-819 (rs1800871; C&gt;T), IL10-1082 (rs1800896; A&gt;G), IL10-2763 (rs6693899; A&gt;C), IL10-2849 (rs6703630; A&gt;G) and IL10-3575 (rs1800890; A&gt;T). In the meta-analysis, we used odds ratio (OR) and 95% confidence interval (CI) to evaluate the association of IL10-1082, IL10-819 and IL10-592 polymorphisms and MS. We found a positive association of MS with the IL10-1082 SNP in genotypic comparison (AG+GG vs. AA) (OR = 1.23; 95% CI = 1.01–1.51; p = 0.04). Our search did not find any article relating polymorphisms in the IL10 gene with ALS. Therefore, our analysis indicates a possible association of IL10 gene SNPs in the development and progression of MS.

TRIPTERIGIUM WILFORDII EXTRACT AS A REGULATOR OF SPONTANEOUS AND IN VITRO STIMULATED CYTOKINE PRODUCTION IN ATOPIC DERMATITIS

Background. The analysis of traditional, primarily Chinese, medicine is the subject of close attention of modern medicinal substances developers. One of the important, frequently used plant substrates is Tripterygium Wilfordii, which is used as an antiinflammatory agent, including for the treatment of skin diseases. However, the mechanisms of action of this agent, particularly in such a widespread disease as atopic dermatitis, still need to be investigated. Aim. To study of the effect of Tripterygium Wilfordii extract on in vitro cytokine production in normal and atopic dermatitis patients. Materials and Methods. In a culture of the effect of Tripterygium Wilfordii extract on spontaneous and phytohemagglutinininduced synthesis of IL-2, IL-4, IL-6, IL-10, IL-17, TNFα, IFNγ was evaluated in mononuclear cells culture obtained from healthy donors and patients with atopic dermatitis using enzyme immunoassay test systems Cloud-Clone Corp. (CCC, Wuhan) Results. It was found that the level of spontaneous cytokine synthesis in patients with atopic dermatitis was significantly increased, while the response to the mitogenic stimulus was sharply reduced, absent or inverted. Tripterygium Wilfordii extract in all samples, except for intact cells of healthy donors, showed a pronounced inhibitory effect. Conclusions. Under in vitro conditions, cultured mononuclear cells of patients with atopic dermatitis produce a greater amount of IL-2, IL-4, IL-6, IL-10, IL-17, TNFα, IFNγ, which indicates the activation of the cytokine function. Tripterygium Wilfordii extract has a pronounced inhibitory effect, especially on activated cells. The use of Tripterygium Wilfordii extracts seems reasonable for promising pharmaceutical substances.

Cytokines in Liver Cirrhosis (Their Importance in Assessing Activity and Decompensation)

Aim: studying the role of pro-inflammatory and anti-inflammatory cytokines in the pathogenesis of liver cirrhosis progression.Materials and methods. The material of the study was the data of clinical-instrumental, biochemical, virological studies of 109 patients with liver cirrhosis of various etiologty, who were hospitalized in the clinic of the Institute of Gastroenterology (Dushanbe, Republic of Tajikistan). The diagnosis of the underlying disease was established according to the clinical recommendations of the Russian Society for the Study of the Liver and the Russian Gastroenterological Association for the diagnosis and treatment of liver fibrosis and cirrhosis and their complications (2021); decompensated liver cirrhosis was established according to the 1996 Child — Pugh classification. The age of the patients ranged from 17 to 79 years (36.9 ± 0.8 years), there were 55 men and 54 women.Results. Among the examined patients, compensated liver cirrhosis (Class A) according to Child — Pugh was detected in 18 persons, subcompensated (Class B) — in 14, decompensated (Class C) — in 77. The study of the content of pro-inflammatory and anti-inflammatory cytokines in the blood serum of patients with Class A liver cirrhosis showed, that levels of tumour necrosis factor alpha (TNF-α), interleukin-2, interleukin-6 were statistically higher compared to healthy individuals, while the concentration of anti-inflammatory interleukin-10 was lower (30.7 ± 4.7 pg/mL) in comparison with the control group. In patients with Class B liver cirrhosis, the level of TNF-α increased to 75.0 ± 4.5 pg/mL (p &lt; 0.001), interleukin-2 — to 328.7 ± 23.9 pg/mL (p &lt; 0.05), and interleukin-6 — to 95.4 ± 7.7 pg/mL (p &lt; 0.001). Serum interleukin-10 decreased compared with the control group (23.1 ± 2.8 pg/mL; p &gt; 0.05). At the decompensated stage of Class C cirrhosis, a huge release of pro-inflammatory cytokines occurs — the content of TNF-α increases by 80 times, of interleukin-2 — by more than 60 times, аs for interleukin-10, its content is progressively reduced.Conclusion. In liver cirrhosis, there is a significant disruption in the synthesis of pro-inflammatory cytokines, which is manifested by a sharp increase in the content of TNF-α, interleukin-2 and interleukin-6. High levels of proinflammatory cytokines in blood serum in liver cirrhosis correlate with the activity and degree of decompensation, which indicates their important role in the pathogenesis and progression of the pathological process.

Pimecrolimus protects neuron-like SH-SY5Y cells against anti-inflammatory and anti-oxidant effects of both microglial secretome and hydrogen peroxide.

Calcineurin inhibitors have been found to exhibit a preventive role against neuroinflammation, which represents a crucial underlying mechanism in neurodegenerative diseases (ND). Additionally, they possess suppressive effects on the activation of apoptotic pathways, which constitute another mechanism underlying such diseases. Given that pimecrolimus, a calcineurin inhibitor, impedes the synthesis of pro-inflammatory cytokines, such as interleukin (IL)-2, IL-4, and IL-10, and influences apoptotic processes, it is noteworthy to test its potential neuroprotective properties. Thus, the objective of this investigation was to assess the potential protective effects of pimecrolimus against the degenerative consequences of both microglial secretomes and hydrogen peroxide (H2O2), an oxidant agent. The survival rates of HMC3 microglia cells, neuron-like differentiated SH-SY5Y (d-SH-SY5Y) cells, and their co-culture were determined using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) method. Furthermore, the levels of pro-inflammatory cytokines IL-1β and IL-6, and anti-inflammatory cytokine IL-10 were measured using ELISA kits, besides total antioxidant and oxidant capacities in conditioned media of cells. Additionally, the effect of pimecrolimus on neurite length in these cell groups was evaluated through morphological observations. This study revealed, for the first time, that pimecrolimus exerts preventive effects on neurodegenerative processes by virtue of its anti-inflammatory and -antioxidant activities. It holds promise as a potential treatment option for ND.

Interferon induction by STING requires its translocation to the late endosomes.

To combat microbial infections, mammalian cells use a variety of innate immune response pathways to induce synthesis of anti-microbial proteins. The cGAS/STING pathway recognizes cytoplasmic viral or cellular DNA to elicit signals that lead to type I interferon and other cytokine synthesis. cGAMP, synthesized by DNA-activated cGAS, activates the ER-associated protein, STING, which oligomerizes and translocates to other intracellular membrane compartments to trigger different branches of signaling. We have reported that, in the ER, EGFR-mediated phosphorylation of Tyr245 of STING is required for its transit to the late endosomes, where it recruits and activates the transcription factor IRF3 required for IFN induction. In the current study, we inquired whether STING Tyr245 phosphorylation per se or STING's location in the late endosomes was critical for its ability to recruit IRF3 and induce IFN. Using pharmacological inhibitors or genetic ablation of proteins that are essential for specific steps of STING trafficking, we demonstrated that the presence of STING in the late endosomal membranes, even without Tyr245 phosphorylation, was sufficient for IRF3-mediated IFN induction.

Current affairs in the use of medical ozone. Biological effects. Mechanisms of action

Introduction. Oxygen-ozone therapy stands as a medically endorsed practice confirmed by numerous international clinical studies. Various authors have illustrated the beneficial clinical outcomes of ozone therapy in terms of its capacity to regulate redox balance, cellular inflammatory responses, and adaptation to ischemia/reperfusion processes. Ozone therapy extends to encompass a range of viral infections, inflammatory disorders, and degenerative ailments, used as both monotherapy and as an adjunct to unified conventional therapies. Material and methods. Narrative literature review study. Bibliographic search was conducted using the PubMed, Hinari, and SpringerLink databases, as well as the National Center of Biotechnology Information and Medline. Articles published between 1990 and 2022 were selected using various combinations of keywords, including “ozone”, “ozone therapy”, “mechanisms of ozone action”, “biological effects of ozone”, “antioxidant effect”, “anti-inflammatory effect” and “immunomodulatory effect.” Information regarding ozone’s mechanisms of action was identified and processed. Following the database information processing and search criteria, a total of 475 full-text articles were found. The final bibliography consists of 52 relevant sources that were deemed representative of the materials published on the topic of this synthesis article. Results. The effects of ozone on oxygen metabolism are explained by changes in the rheological properties of blood, including inhibition of erythrocyte aggregation and stimulation of 2,3-diphosphoglycerate in erythrocytes, favoring the transport and delivery of oxygen to tissues while facilitating the substantial elimination of nitric oxide and increasing blood flow. Intracellular triatomic oxygen enhances the oxidative carboxylation of pyruvate, stimulating ATP production, which also contributes to reducing peripheral vascular resistance. Conclusions. Ozone generates a moderate oxidative stress. Yet, it can set off several beneficial biochemical mechanisms that reactivate both the intra- and extracellular antioxidant systems and reverse chronic oxidative stress in various inflammatory and degenerative processes. Ozone induces a mild activation of the immune system by triggering neutrophil activation and stimulating the synthesis of certain cytokines (IL-2, TNF-α, IL-6, and IFN-γ), thereby initiating a complete cascade of immune responses. Ozone therapy yields the following biological reactions: optimization of blood circulation and oxygen delivery to ischemic tissue, regulation of cellular antioxidant enzymes, initiation of a slight immune system activation, and enhancing the release of growth factors.

The Influence of KE and EW Dipeptides in the Composition of the Thymalin Drug on Gene Expression and Protein Synthesis Involved in the Pathogenesis of COVID-19.

Thymalin is an immunomodulatory drug containing a polypeptide extract of thymus that has demonstrated efficacy in the therapy of acute respiratory distress syndrome and chronic obstructive pulmonary disease, as well as in complex therapy related to severe COVID-19 in middle-aged and elderly patients.. KE and EW dipeptides are active substances of Thymalin. There is evidence that KE stimulates cellular immunity and nonspecific resistance in organisms, exerting an activating effect on macrophages, blood lymphocytes, thymocytes, and neutrophils, while EW reduces angiotensin-induced vasoconstriction and preserves endothelium-dependent vascular relaxation by inhibiting ACE2, the target protein of SARS-CoV-2. However, the mechanism of the immunomodulatory action of Thymalin, KE, and EW during COVID-19 remains unclear. To identify the potential mechanism of action underlying the immunomodulatory activity of Thymalin and its active components, EW and KE dipeptides, we assessed inflammatory response in the context of COVID-19. Interactions between EW and KE dipeptides and double-stranded DNA (dsDNA) were investigated by molecular modeling and docking using ICM-Pro. Analysis of the possible effect of EW and KE dipeptides on gene expression and protein synthesis involved in the pathogenesis of COVID-19 was conducted through the use of bioinformatics methods, including a search for promoter sequences in the Eukaryotic Promoter Database, the determination of genes associated with the development of COVID-19 using the PathCards database of human biological pathways (pathway unification database), identification of the relationship between proteins through cluster analysis in the STRING database ('Search Tool for Retrieval of Interacting Genes/Proteins'), and assessment of the functional enrichment of protein-protein interaction (PPI) using the terms of gene ontology (GO) and the Markov cluster algorithm (MCL). After that, in vitro studying of a lipopolysaccharide (LPS)-induced model of inflammation using human peripheral blood mononuclear cells was performed. ELISA was applied to assess the level of cytokines (IL-1β, IL-6, TNFα) in the supernatant of cells with or without the impact of EW and KE peptides. Blood samples were obtained from four donors; for each cytokine, ELISA was performed 2-4 times, with two parallel experimental or control samples for each experiment (experiments to assess the effects of peptides on LPS-stimulated cells were repeated four times, while additional experiments with unstimulated cells were performed two times). Using molecular docking, GGAG was found to be the best dsDNA sequence in the classical B-form for binding the EW dipeptide, while GCGC is the preferred dsDNA sequence in the curved nucleosomal form for the KE dipeptide. Cluster analysis revealed that potential target genes for the EW and KE peptides encode the AKT1 and AKT2 proteins involved in the development of the cytokine storm. The specific targets for the EW peptide are the ACE2 and CYSLTR1 genes, and specific target for the KE peptide is the CHUK gene. Protein products of the ACE2, CYSLTR1, and CHUK genes are functionally associated with IL-1β, IL-6, TNF-α, IL-4, and IL-10 cytokines. An in vitro model of an inflammatory reaction demonstrated that Thymalin and EW and KE dipeptides reduced the synthesis of IL-1β, IL-6, and TNF-α cytokines in human peripheral blood mononuclear cells by 1.4-6.0 times. The immunomodulatory effect of Thymalin under the inflammatory response conditions in COVID-19 is based on the potential ability of its active components, EW and KE dipeptides, to regulate protein synthesis involved in the development of the cytokine storm.

Paroxetine's effect on the proinflammatory cytokine stimulation and intracellular signaling pathways in J774.2 cells.

Paroxetine is extensively utilized in the management of depressive and anxious conditions. Paroxetine works by increasing serotonin levels in nerve cells in the brain. However, limited information is available regarding the direct effects of paroxetine on macrophage cells. Macrophages are a type of leukocytes involved in the body's immune response, playing a crucial role in combating infections. The impact of paroxetine on macrophages has been explored in research, although a comprehensive understanding is still pending. This study aimed to research the potential of administering paroxetine to J774.2 macrophage cells to stimulate the release of GM-CSF, TNF-α, IL-12p40, and IL-6 cytokines. Additionally, we examined the mechanisms of action of paroxetine on the p38 signaling pathway, which is involved in cytokine production, and the PI3K pathway, which is an important mechanism in intracellular signaling. Our findings revealed that paroxetine induced an inflammatory response in macrophages by promoting cytokine synthesis in a non-lipopolysaccharide (LPS) environment. We observed that paroxetine triggered the inflammatory response through the PI3K signaling pathway while suppressing the p38 signaling pathway.

Role of myeloid-derived suppressor cells in prediction of the effectiveness of biologics in children with psoriasis

Psoriasis is a chronic inflammatory skin disease characterized by increased proliferation of epidermal cells, impaired keratinization, and an inflammatory reaction in the dermis due to activation of T-lymphocytes and synthesis of proinflammatory cytokines. Pathophysiology of psoriasis is associated not only with activation of proinflammatory reactions, but also with decreased anti-inflammatory functions of immunosuppressive cells. It is known that Treg, Breg and MDSCs do not perform their classical homeostatic functions in psoriasis. In recent years, there have been more and more cases of developing resistance to ongoing therapy with genetically engineered biological drugs (GEBD) in childhood, requiring replacement or discontinuation of the drug. The aim of our work was to estimate the content of MDSCs subpopulations and their functional activity in the peripheral blood of children with psoriasis at different effectiveness of biotherapeutic drugs. We examined 110 children with psoriasis vulgaris before the appointment of biologics, at 16 and 52 weeks of therapy with adalimumab, etanercept and ustikinumab, aged 6 to 18 years. Сomparison group consisted of 34 healthy children. The effectiveness of therapy was assessed by the achievement of PASI 75 by one year of therapy. Contents of myeloid-derived suppressor cells (MDSCs) and their subpopulations, and the activity of arginase-1 were assessed by multicolor flow cytometry. An increased content of MDSCs was found in children with psoriasis against the comparison group (p = 0.000). Analysis of the effectiveness of biologics in children with psoriasis, according to PASI, showed a significant reduction in disease severity in the group of patients with good effect, both at week 16 of therapy (p = 0.000) and by one year (p = 0.017). In the group of patients with good effect of biological therapy, percentage of total MDSCs population was higher, both before start of treatment and by 52nd week of therapy (p 0.01). Children with psoriasis showed increased immunosuppressive function of MDSCs by arginase-1 activity versus the comparison group (p = 0.000). The arginase-1 activity in patients with psoriasis at the stage of disease regression (PASI 10) was significantly increased relative to children in progressive stage of psoriasis (PASI10; p = 0.001). Thus, the content of MDSCs and their suppressive activity in children with psoriasis is an informative efficiency predictor of the biological drugs. Fading of biotherapy effect after the induction course is accompanied by decreased number of MDSCs and their functional activity.

The mechanisms related to fibroblasts in burn surface.

Mesenchymal stem cells (MSCs) can promote burn wound healing, skin appearance, and function recovery by promoting the differentiation and migration of fibroblasts of a wound. The burn environment can activate the autophagy of MSCs. However, it is not clear whether this autophagy can affect the proliferation and migration of fibroblasts. In this study, pretreated MSCs with rapamycin and 3-methyladenine modulated autophagy and co-cultured with fibroblasts of burn. Cell migration was detected by immunofluorescence chemical staining. Western blot analysis and enzyme-linked immunosorbent assay were performed to detect 2,3-Dioxygenase (IDO), cytokine synthesis inhibitory factor 10 (IL-10), cytokine synthesis inhibitory factor 6 (IL-6), prostaglandin E2 (PGE2), transforming growth factor beta 1 (TGF-β1) proteins levels, and the autophagy proteins p62 and microtubule-associated protein LC3-II/I. We demonstrated that autophagy regulates MSCs survival and proliferation in burn wound transplants and found that autophagy inhibition with 3-methyladenine reduced MSCs-mediated, fibroblast proliferation and migration in burn environment. However, rapamycin-induced autophagy had the opposite effect and increased the TGF-β1 expression. Therefore, we speculate that MSCs may promote fibroblast proliferation and migration by secreting TGF-β1 via the AKT/mTOR (RAC-alpha serine/threonine-protein kinase/mammalian target of rapamycin) pathway. Autophagy of MSCs regulates burn wound fibroblast proliferation and migration by affecting TGF-β1 and prostaglandin E2 production adjacent to MSCs transplanted on the burn wound. The results of this study provide a potential strategy for promoting MSCs treatment of burns.

Modulating the m6A Modified Transcription Factor GATA6 Impacts Epithelial Cytokines in Acute Lung Injury.

The methylation of the N6-position of adenosine (m6A) has been found to be associated with the inflammatory response. We hypothesize that m6A modification plays a role in the inflammation of airway epithelial cells during lung inflammation. However, the precise changes and functions of m6A modification in airway epithelial cells in acute lung injury (ALI) are not well understood. Here we report that RNA Methyltransferase 3 (METTL3)-mediated m6A of GATA6 mRNA inhibits ALI and the secretion of pro-inflammatory cytokine in airway epithelial cells. The expression of METTL3 and m6A levels decrease in lung tissues of ALI-mice. In co-cultures, peripheral blood monocytes secreted tumor necrosis factor α (TNFα) which reduces METTL3 and m6A levels in human bronchial epithelial cell line (BEAS-2B). Knockdown of METTL3 promotes IL-6 and TNFα release of BEAS-2B cells. Conversely, overexpression of METTL3 increases total RNA m6A level and reduces the levels of pro-inflammatory cytokines TNFα, transforming growth factor-β (TGFβ), and thymic stromal lymphopoietin (TSLP). Elevating METTL3 in mouse lungs prevented LPS-induced ALI and reduced the synthesis of pro-inflammatory cytokines. Mechanistically, sequencing and functional analysis show that METTL3 catalyzes m6A in the 3'UTR of GATA6 read by YTH N6-Methyladenosine RNA Binding Protein 2 (YTHDF2) and triggers mRNA degradation. GATA6 knockdown rescues TNFα induced inflammatory cytokine secretion of epithelial cells, indicating that GATA6 is a main substrate of METTL3 in airway epithelial cells. Overall, this study provides evidence of a novel role for METTL3 in the inflammatory cytokine release of epithelial cells and provide an innovative therapeutic target for ALI.