Cytokines TNF-alpha Research Articles

Exploring the Potential of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 as Promising Psychobiotics Using SHIME.

Psychobiotics are probiotics that have the characteristics of modulating central nervous system (CNS) functions or reconciled actions by the gut-brain axis (GBA) through neural, humoral and metabolic pathways to improve gastrointestinal activity as well as anxiolytic and even antidepressant abilities. The aim of this work was to evaluate the effect of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 on the gut microbiota of mildly anxious adults using SHIME®. The protocol included a one-week control period and two weeks of treatment with L. helveticus R0052 and B. longum R0175. Ammonia (NH4+), short chain fatty acids (SCFAs), gamma-aminobutyric acid (GABA), cytokines and microbiota composition were determined. Probiotic strains decreased significantly throughout the gastric phase. The highest survival rates were exhibited by L. helveticus R0052 (81.58%; 77.22%) after the gastric and intestinal phase when compared to B. longum (68.80%; 64.64%). At the genus level, a taxonomic assignment performed in the ascending colon in the SHIME® model showed that probiotics (7 and 14 days) significantly (p < 0.005) increased the abundance of Lactobacillus and Olsenella and significantly decreased Lachnospira and Escheria-Shigella. The probiotic treatment (7 and 14 days) decreased (p < 0.001) NH4+ production when compared to the control period. For SCFAs, we observed after probiotic treatment (14 days) an increase (p < 0.001) in acetic acid production and total SCFAs when compared to the control period. Probiotic treatment increased (p < 0.001) the secretion of anti-inflammatory (IL-6 and IL-10) and decreased (p < 0.001) pro-inflammatory cytokines (TNF-alpha) when compared to the control period. The gut-brain axis plays an important role in the gut microbiota, producing SCFAs and GABA, stimulating the production of anti-anxiety homeostasis. The signature of the microbiota in anxiety disorders provides a promising direction for the prevention of mental illness and opens a new perspective for using the psychobiotic as a main actor of therapeutic targets.

Targeting the Endoplasmic Reticulum Stress-Linked PERK/GRP78/CHOP Pathway with Magnesium Sulfate Attenuates Chronic-Restraint-Stress-Induced Depression-like Neuropathology in Rats.

Magnesium sulfate has demonstrated marked neuroprotection in eclampsia, hypoxia, stroke, and post-traumatic brain injury rodent models. However, its potential impact against chronic-restraint-stress (CRS)-induced depression-like neuropathology and associated alterations in endoplasmic reticulum (ER) stress have not been adequately examined. The present study aimed to investigate the neuroprotective potential of magnesium sulfate in a rat model of CRS-triggered depression-like behavioral disturbance and the underlying molecular mechanisms. Herein, CRS was induced by placing rats into restraining tubes for 6 h/day for 21 days and the animals were intraperitoneally injected with magnesium sulfate (100 mg/kg/day) during the study period. After stress cessation, the depression-like behavior was examined by the open-field test, sucrose preference test, and forced swimming test. The present data demonstrated that CRS triggered typical depression-like behavioral changes which were confirmed by the Z-normalization scores. Mechanistically, serum circulating corticosterone levels spiked, and the hippocampi of CRS-exposed animals demonstrated a significant decline in serotonin, norepinephrine, and dopamine neurotransmitters. At the molecular level, the hippocampal pro-inflammatory TNF-alpha and IL-1β cytokines and the oxidative stress marker 8-hydroxy-2'-deoxyguanosine (8-HG) increased in stressed animals. In tandem, enhancement of hippocampal ER stress was evidenced by the activation of iNOS/PERK/GRP78/CHOP axis seen by increased protein expression of iNOS, PERK, GRP78, and CHOP signal proteins in the hippocampi of stressed rats. Interestingly, magnesium sulfate administration attenuated the depression-like behavioral outcomes and the histopathological changes in the brain hippocampi. These favorable actions were driven by magnesium sulfate's counteraction of corticosterone spike, and hippocampal neurotransmitter decline, alongside the attenuation of neuroinflammation, pro-oxidation, and ER stress. In conclusion, the current results suggest the promising neuroprotective/antidepressant actions of magnesium sulfate in CRS by dampening inflammation, ER stress, and the associated PERK/GRP78/CHOP pathway.

Alterations in heart rate variability and pro-inflammatory cytokine TNF-alpha in individuals with panic disorder

Based on the autonomic flexibility–neurovisceral integration model, panic disorder (PD) has been associated with a generalized proinflammatory state and lower cardiac vagal tone. The heart rate variability (HRV) provides an index of cardiac autonomic function and reflects the parasympathetic innervation to the heart regulated by the vagus nerve. The aim of this study was to explore the heart rate variability, pro-inflammatory cytokines and their associations in individuals with PD. Short-term HRV with time and frequency domain indices as well as pro-inflammatory cytokines Interleukin-6 (IL-6) and Tumor Necrosis Factor alpha (TNF-alpha) were assessed in seventy individuals with PD (mean age: 35.64 ± 14.21 years) and thirty-three healthy controls (mean age: 38.33 ± 14.14 years). Individuals with PD showed significantly lower HRV in the time and frequency domain parameters during a short-term resting condition. A lower TNF-alpha concentration could be observed in individuals with PD in comparison to healthy controls, but no differences in IL-6. Furthermore, the HRV parameter absolute power in the low-frequency band 0.04–0.15 Hz (LF) predicted TNF-alpha concentrations. In conclusion, a lower cardiac vagal tone, decreased adaptive autonomic nervous system (ANS), and higher pro-inflammatory cytokine state could be observed in individuals with PD compared to healthy controls.

Neuroimmune interactions with binge alcohol drinking in the cerebellum of IL-6 transgenic mice

The neuroimmune system of the brain, which is comprised primarily of astrocytes and microglia, regulates a variety of homeostatic mechanisms that underlie normal brain function. Numerous conditions, including alcohol consumption, can disrupt this regulatory process by altering brain levels of neuroimmune factors. Alcohol and neuroimmune factors, such as proinflammatory cytokines IL-6 and TNF-alpha, act at similar targets in the brain, including excitatory and inhibitory synaptic transmission. Thus, alcohol-induced production of IL-6 and/or TNF-alpha could be important contributing factors to the effects of alcohol on the brain. Recent studies indicate that IL-6 plays a role in alcohol drinking and the effects of alcohol on the brain activity following the cessation of alcohol consumption (post-alcohol period), however information on these topics is limited. Here we used homozygous and heterozygous female and male transgenic mice with increased astrocyte expression of IL-6 to examined further the interactions between alcohol and IL-6 with respect to voluntary alcohol drinking, brain activity during the post-alcohol period, IL-6 signal transduction, and expression of synaptic proteins. Wildtype littermates (WT) served as controls. The transgenic mice model brain neuroimmune status with respect to IL-6 in subjects with a history of persistent alcohol use. Results showed a genotype dependent reduction in voluntary alcohol consumption in the Drinking in the Dark protocol and in frequency-dependent relationships between brain activity in EEG recordings during the post-alcohol period and alcohol consumption. IL-6, TNF-alpha, IL-6 signal transduction partners pSTAT3 and c/EBP beta, and synaptic proteins were shown to play a role in these genotypic effects.

Lyophilized Extracellular Vesicles from Adipose-Derived Stem Cells Increase Muscle Reperfusion but Degrade Muscle Structural Proteins in a Mouse Model of Hindlimb Ischemia-Reperfusion Injury.

Ischemia-reperfusion (I/R) injury is a complication impacting multiple organs and tissues in clinical conditions ranging from peripheral arterial disease to musculoskeletal trauma and myocardial infarction. Stem cell-derived extracellular vesicles (EVs) may represent one therapeutic resource for preventing the tissue damage associated with I/R injury. Here we tested the hypothesis that lyophilized extracellular vesicles derived from adipose stem cells could serve as an "off-the-shelf" treatment modality for I/R injury in a mouse hindlimb ischemia model. Ischemia was induced for 90 min using a rubber band tourniquet and extracellular vesicles (0, 50, or 100 µg) administered via tail vein injection immediately prior to reperfusion. Perfusion was measured prior to, during, and after ischemia using laser Doppler imaging. Serum and tissue were collected 24 h after reperfusion. Mass spectrometry (MS)-based proteomics was used to characterize the EV cargo and proteins from the ischemic and non-ischemic hindlimb. Inflammatory cytokines were measured in muscle and serum using a multiplex array. Results indicate that EVs significantly increase reperfusion and significantly increase expression of the anti-inflammatory factor annexin a1 in skeletal muscle; however, the increased reperfusion was also associated with a marked decrease in muscle structural proteins such as dystrophin, plectin, and obscurin. Circulating inflammatory cytokines TNF-alpha and IL-6 were increased with EV treatment, and serum TNF-alpha showed a significant, positive correlation with reperfusion level. These findings suggest that, while EVs may enhance reperfusion, the increased reperfusion can negatively impact muscle tissue and possibly remote organs. Alternative approaches, such as targeting mitochondrial permeability, may be more effective at mitigating I/R injury.

Evaluation of Antibacterial, Antioxidant and Anti-inflammatory Properties of Methanol Extract of Varthemia iphionoides

Medicinal plants represent an essential source of active biological compounds. The current study examined the methanol extract of V. iphionoides for its antibacterial, antioxidant, and antiinflammatory activities. The antibacterial activity was evaluated against six bacterial strains (Staphylococcus xyloses, Escherichia coli, Klebsiella oxytoca, Enterobacter aerogenes, Achromabacter xylosoxidans, and Pseudomonas aerugenosa). V. iphionoides extract showed good antibacterial activity against Staphylococcus xylosus followed by, pseudomonas aeruginosa and Klebsiella oxytoca, and the minimum inhibitory concentrations (MICs) were 33, 33, and 11 µg/ml, respectively. However, the extract did not show any antibacterial activity against the other bacterial strains. The antioxidant activity of the plant extract was evaluated using ABTS and DPPH radical scavenging assays, and the Trolox equivalents were 93±0.31 and 84.8±0.28 mg Trolox/g plant extract, respectively. Anti-inflammatory activity in LPS-induced mice was investigated by measuring the concentration of pro-inflammatory cytokines TNF-alpha, IL 1beta, and antioxidant enzyme catalase and glutathione peroxidase. The results demonstrated that the highest concentration of V. iphionoides extract was the most efficient. Anti-inflammatory activity was exhibited by reducing the concentrations of pro-inflammatory TNF-α and IL 1 beta and reducing the concentrations of antioxidant enzyme catalase and glutathione peroxidase compared to the negative control group.

Effect of a slow-paced breathing with heart rate variability biofeedback intervention on pro-inflammatory cytokines in individuals with panic disorder - A randomized controlled trial

A generalized proinflammatory state has been observed in individuals with panic disorder (PD). There is evidence that slow-paced breathing (SPB) with heart rate variability-biofeedback (HRV-BF) strengthens the nervus vagus with its anti-inflammatory pathway. Therefore, with this randomized controlled trial we aimed to investigate the effect of a four-week SPB with HRV-BF intervention on pro-inflammatory cytokines in people with PD. Fifty-five individuals with PD (mean age: 37.22 ± 15.13 years) were randomly allocated either to SPB-HRV-BF (intervention group) or to HRV-Sham-BF (active control group). SPB-HRV-BF was performed over four weeks while cytokine concentration and HRV during a short-term resting condition were measured before and after intervention. SPB-HRV-BF decreased concentration of Tumor Necrosis Factor alpha (TNF-alpha) (F(1, 53) = 4.396, p ≤ .05, η2 = 0.077) in individuals with PD. In addition, SPB-HRV-BF demonstrated an increase in the HRV-time and frequency domain parameters SDNN, Total Power and LF during short-term resting condition. There was no intervention effect in HRV-Sham-BF group. In conclusion, SBP-HRV-BF as a non-pharmacological treatment may reduce pro-inflammatory cytokine TNF-alpha via the cholinergic anti-inflammatory pathway in individuals with PD. Based on the generalized proinflammatory state in PD, decreasing TNF-alpha is highly beneficial to reduce risk of cardiovascular diseases and metabolic syndrome.

Thymus vulgaris, a natural pharmacy against COVID-19: A molecular review

IntroductionA worldwide pandemic infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of a deadly disease called COVID-19. Interaction of the virus and the Angiotensin converting-enzyme 2 (ACE2) receptor leads to an inflammatory-induced tissue damage. Thymus vulgaris L. (TvL) is a plant with a long history in traditional medicine that has antimicrobial, antiseptic, and antiviral properties. Thymol and Carvacrol are two important biological components in Thyme that have anti-inflammatory, antioxidant, and immunomodulatory properties. This study is a molecular review on the potential effects of TvL and its active compounds on SARS-COV2 infection. MethodThis is a narrative review in which using PubMed, Scopus, ISI, Cochrane, ScienceDirect, Google scholar, and Arxiv preprint databases, the molecular mechanisms of therapeutic and protective effects of TvL and its active compounds have been discussed regarding the molecular pathogenesis in COVID-19. ResultsThyme could suppress TNF-alpha, IL-6, and other inflammatory cytokines. It also enhances the anti-inflammatory cytokines like TGF-beta and IL-10. Thyme extract acts also as an inhibitor of cytokines IL-1-beta and IL-8, at both mRNA and protein levels. Thymol may also control the progression of neuro-inflammation toward neurological disease by reducing some factors. Thyme and its active ingredients, especially Thymol and Carvacrol, have also positive effects on the renin-angiotensin system (RAS) and intestinal microbiota. ConclusionsAccordingly, TvL and its bioactive components may prevent COVID-19 complications and has a potential protective role against the deleterious consequences of the disease.

Age, Sex, Metabolic and Pharmacologic Factors May Predict Nonresponse Status to Rheumatoid Arthritis Therapies.

A real challenge for patients with rheumatoid arthritis (RA) and rheumatologists is primary nonresponse status (PNRS) or secondary nonresponse status (SNRS) to various therapies. Despite their detrimental influence on patient life quality, PNRS and SNRS have no accurate definition and no early predictive criteria for their development exist. Patients with RA under 40 years of age are rare, hence PNRS and SNRS data for this age group are scarce. This study examined the PNRS and SNRS according to sex, age, BMI, therapy type, and duration. Retrospectively, 115 patients with RA having PNRS and/or SNRS were stratified by age (22-39, 40-59, and 60-81). The association between body mass index (BMI), proinflammatory cytokines inhibitors, JAK inhibitors, and TNF-alpha inhibitors, sex, age, and PNRS and SNRS was examined. All three proinflammatory cytokine inhibitors (rituximab, tocilizumab, and abatacept) were associated with PNRS and SNRS in women with a high BMI aged 40-59 years. Abatacept-related PNRS and SNRS was significant in women with normal BMI aged 60-81 years. Adalimumab, infliximab, and golimumab affected SNRS differently in women with normal BMI aged 22-39 years and women with high BMI aged 60-81 years. Etanercept and infliximab were associated with SNRS status in men with high-BMI aged 40-59 years. PNRS and SNRS development in patients with RA is significantly influenced by age, sex, and BMI, but most importantly is closely and differentially related to therapy type and duration.

TNF-alpha and IL-17 cytokine levels in Brazilian patients with ankylosing spondylitis after anti-TNF therapy

TNF-alpha and IL-17 cytokine levels in Brazilian patients with ankylosing spondylitis after anti-TNF therapy

Functionality of a Peripheral Blood Derived Microglia-like Cell System for Drug Discovery to Treat Sickle Cell Related Chronic Pain

Background: Abnormal microglial activation is emerging as a clinically promising drug target to treat chronic pain in sickle cell disease (SCD). But research involving microglia has been stymied by difficulty in obtaining human CNS microglia and differences between human and animal neurobiology. To address such limitations, we have developed an in vitro culture system to induce human microglia-like cells (MLC) from peripheral blood mononuclear cells. In this study, we further validated the functionality of the cultured peripheral blood derived microglia-like cell (PB-MLC) and evaluate the pathological characteristics captured in the PB-MLC model system, which is an important requirement for disease model systems used in drug discovery. Methods: Human peripheral blood mononuclear cells were cultured with human GM-CSF (10 ng/ml), human IL-34 (100 ng/ml), TGF-β2 (2 ng/mL) and cholesterol (1.5 µg/mL) for 7 days to induce Human peripheral blood derived microglia-like cells (HPB-MLC). HPB-MLC were morphologically analyzed by phase contrast microscopy, phenotyped by immunofluorescence staining with anti-TMEM119, P2RY12, and Iba1 antibodies. To validate that the HPB-MLC culture system recapitulates important pathological features, we compared the microglial morphology, activation states and phagocytic activity of HPB-MLC from individuals with SCD with or without chronic pain, individuals with headache and normal donors without pain: Cells were stimulated with 100 ng/ml LPS for 24 hours, and secretion of TNF-alpha, IL-1beta, and IL-6 was measured by ELISA. The microglial phagocytic function of HPB-MLC was determined by assessing the uptake of fluorescent-labeled Zymosan particles with fluorescent microscopy and imaging flow cytometry. The model system was also validated by comparing cultured PB-MLC to CNS derived microglia cells using Sprague-Dawley rats: Rat brain derived microglia (BDM) and rat peripheral blood monocytes were isolated and then cultured with murine IL-34 (100 ng/ml), murine GM-CSF (10 ng/ml). TGF-β2 (2 ng/mL), and cholesterol (1.5 µg/mL) for 7 days. The cultured cells were morphologically analyzed as above; phenotyped with anti-TMEM119, P2RY12, and Iba1 antibodies. Rat BDM, rat PB-MLC and HPB-MLC were treated with 100 ng/ml LPS with or without minocycline (2.5, 5, 10, 25, 50 μg/mL) for 24 hours; TNF-alpha secretion was measured by ELISA. Results: Rat PB-MLC resemble BDM morphologically and express microglia specific markers. HPB-MLC express microglia specific markers (TMEM119, P2RY12, Iba1), exhibit microglial phagocytic activity. HPB-MLC retain the patient pain phenotype: Cultures from donors without chronic pain show more ramified morphology and lower phagocytic activity than cultures from individuals with chronic pain (Figure 1); HPB-MLC can be activated by LPS, causing microglial deramification and increased the expression of microglial activation marker CD68. HPB-MLC from patients with pain are more easily activated and secrete more inflammatory cytokines in response to LPS; SCD pain and headache samples exhibit different pro-inflammatory cytokine profile. To evaluate the possibility of using the PB-MLC model system to screen compounds to inhibit microglia activation, we tested PB-MLC cells with the microglial inhibitor, minocycline. The compound significantly suppressed the release of proinflammatory cytokine TNF-alpha from LPS-induced activated PB-MLC in a dose-dependent manner (Figure 2). Conclusions: We have developed and validated a microglial-like cell culture system from the peripheral blood of patients with sickle cell disease that retains the patient pain phenotype. HPB-MLC are physiologically functional and exhibit mature microglial features, including LPS-induced activation, microglial specific morphological transition, microglial phagocytic activity, and drug response to a well-studied microglia inhibitor, minocycline. As a cost-effective, efficient, and patient-specific microglia model, HPB-MLC culture system can be applied to identify target compounds that normalize microglia activation, and potentially treat chronic pain. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Development of a High-Throughput Drug Screening System Based on Human Peripheral Blood Derived Microglia-like Cells from Sickle Cell Patients with Chronic Pain

Background: Targeting microglia activation is a therapeutic strategy and a clinically promising drug target to treat microglia involved diseases, such as chronic pain, neurodegenerative disorders. Despite the interest in studying microglia as drug targets, drug discovery has been stymied by species related differences and lack of healthy human CNS microglia in the current models, Importantly, the current model systems do not faithfully recapitulate pathological characteristics of patients or disease processes, leading to the failure of many drug candidates in clinical trials. To address these shortcomings, we developed an efficient suspension culture system to generate peripheral blood derived microglia like cells that can express molecular markers, functional characteristics like true microglia and preserved patient-specific phenotype. As a novel, cost-effective, efficient, and patient-specific microglia model, the system can be applied to screen for compounds that reduce microglia hyperactivity, thereby identifying promising pharmacologic agents to be developed to treat chronic pain. Methods: Human peripheral blood mononuclear cells obtained from patients with sickle cell disease (SCD) were cultured in a suspension culture system for 6 days to generate high-throughput assay-ready human microglia-like cells (HPB-MLC) with human GM-CSF (10 ng/ml), human IL-34 (100 ng/ml), TGF-β2 (2 ng/mL). The microglial phagocytic function of HPB-MLC was determined by assessing the uptake of fluorescent-labeled Zymosan particles with imaging flow cytometry. Microglial activation of HPB-MLC was induced with LPS (100 ng/mL) and characterized by increased expression of TNF-alpha and microglial activation marker CD68. To evaluate pharmaceutical blockade of microglial activation, HPB-MLC were treated with 100 ng/ml LPS with or without minocycline (2.5, 5, 10, 25, 50 μg/mL), clopidogrel (1, 2, 4 μM) and MRS2395 (1, 5, 10 μM) for 24 hours. Commercial homogeneous cell-based assay kits based on HTRF (Homogeneous Time Resolved Fluorescence) and AlphaScreen technologies were used to develop the high-throughput screening (HTS) and miniaturizable assay. Results: Peripheral blood mononuclear cells can be efficiently differentiated into microglia-like cells in a suspension culture system. The suspension culture system enables fast and uniform seeding of cells to 384 well microplate for HTS applications. HPB-MLC express microglia specific markers, microglial phagocytic activity, and can be activated by LPS. HPB-MLC retain the patient pain phenotype: HPB-MLC generated from SCD patients with chronic pain exhibited higher phagocytic activity than that from SCD patients without chronic pain (Figure 1). We developed a cell-based, high-throughput screening assay by combination of our patient-specific microglia model and commercial homogeneous, high-throughput assay technologies HTRF and AlphaScreen (Figure 2). Suspension culture allowed 5 times higher cell concentration than adherent culture, which greatly reduced cost of cytokines in culture and produced assay-ready frozen HPB-MLC. To evaluate the possibility of using the HPB-MLC model system to screen compounds to inhibit microglia activation, we tested HPB-MLC cells with the several microglial inhibitors: minocycline, MRS2395, and clopidogrel. All the drugs significantly suppressed the release of proinflammatory cytokine TNF-alpha from LPS-induced activated PB-MLC in a dose-dependent manner. Conclusions: Peripheral blood monocytes can be efficiently differentiated into MLC in a suspension culture system, using for drug screening as a patient-specific microglia model. The differentiation in suspension culture worked very well in higher cell concentration (5 times higher than adherent culture), which greatly reduced cost of cytokines in culture. Assay-ready frozen HPB-MLC as a bank of uniform aliquots can be directly used for HTS applications. HTRF and AlphaScreen technologies were used to replace traditional ELISA assays to detect inflammatory cytokines, providing a highly sensitive, rapid, homogeneous and miniaturizable method to efficiently perform high throughput drug screening. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract 15505: Cardiac-Specific Jarid2 Deficiency, Results in Improved Survival and Myocardial Function Following Experimental Sepsis

Introduction: Myocardial depression and associated hemodynamic collapse are among the major causes of death in severe sepsis. Management of severe sepsis is largely supportive. Hypothesis: Emerging evidence suggests that the transcriptional repressor Jarid2 may play a regulatory role in sepsis; however, the functional role of Jarid2 in sepsis-induced myocardial dysfunction remains uninvestigated. Methods: Using both in in vivo (murine model of cecum ligation and puncture (CLP) in 12-week-old C57-wild type (WT) and cardiac specific Jarid2(knock-out, KO)) and in vitro (lipopolysaccharide, LPS) treatment of cultured murine cardiac myocytes) approaches we determined whether Jarid2 is a putative novel target for sepsis. Results: In response to CLP for 48hrs, WT displayed &gt;60% mortality, survivors developed bacterial peritonitis, a marked inflammatory response (3-5-fold increases in serum and left ventricular (LV) interleukin (IL)-1beta, IL-6 and tumor necrosis factor (TNF)-alpha), myocardial dysfunction (20-35% decreases in dp/dt and echo-derived % ejection fraction (EF) and % fractional shortening (FS)), and a 40-50% decrease in LV Jarid2mRNA and protein compared to WT-sham. Jarid2KO mice showed resistance to CLP-induced bacterial peritonitis resulting in improved cardiac function (10-15% decrease in %EF and %FS), and survival (&gt;90%) despite an exacerbated (10-15-fold increase in LV) inflammatory response compared to WT mice. In adult cultured cardiac myocytes derived from WT and Jarid2KO mice, treatment with the endotoxin LPS (1 microgram/mL) for 24 hrs, resulted in an 8-10-fold and 20-35-fold increase respectively, in the inflammatory cytokines TNF-alpha, IL-1beta and IL-6. Conclusions: These findings reveal that the loss of Jarid2 enhances bacterial clearance and despite an enhanced inflammatory response, resulted in improved myocardial function and survival following CLP-induced sepsis. As such, Jarid2 may be a novel therapeutic target in the treatment of sepsis-induced myocardial dysfunction.

Endothelial dysfunction markers and immune response indices in cosmonauts’ blood after long-duration space flights

Space flight factors are known to cause a malfunction in the human immune system and lead to damage to blood vessels. The hemostatic function of endothelium during space missions and its interaction with human immunity has not been determined so far. In this work, we investigated the markers of endothelial activation and damage (plasma concentrations of soluble thrombomodulin fraction (sTM), von Willebrand factor (vWF), highly sensitive C-reactive protein (hs-CRP)), as well as the level of D-dimer and compared them to the immunological parameters characterizing the state of human humoral and cellular immunity. The immune status of long-duration ISS crewmembers was assessed by whole-blood testing, and comprehensive postflight immune assessment included the analysis of leukocyte distribution. Flow cytometry was applied to determine the absolute counts and the percentage of lymphocyte subsets: B cells (CD19+), T cells (CD3+, CD3+CD4+, CD3+CD8+), NK cells (CD3−CD16+CD56+, CD11b+CD56+), and activated subsets (CD3+CD25+ and CD3+HLA-DR+). The in vitro basal cytokine production was investigated in whole blood cell culture. The cytokines IFN-gamma, IL-1-beta, IL-4, IL-6, IL-10, IL-18, and TNF-alpha were measured in plasma and the 24-h supernatants by a sensitive enzyme-linked immunosorbent assay. A significant increase in the plasma levels of vWF and hs-CRP and a decrease in the concentration of sTM after spaceflights were detected. Divergent changes in the parameters characterizing the state of the immune system were observed. We propose that the changes revealed may lead to an increase in the procoagulant activity of blood plasma, suppression of protein C activation and thrombin inhibition, as well as to an increase in the adhesive-aggregate potential of platelets, especially in case of changes in the rheological characteristics of blood flow during re-adaptation to ground conditions. We also speculate that the immune system might play an important role in vessel damage during long-duration missions.

S3267 A Case of Spontaneous Resolution of Malignant Peritoneal Mesothelioma

Introduction: Malignant peritoneal mesothelioma (MPM) is an aggressive tumor of the peritoneum. Few cases have been reported in the literature describing spontaneous regression possibly related to immune reconstitution. Here in, we report a unique case of peritoneal mesothelioma that regressed over time. Case Description/Methods: A case of 44 y/o female with a history of severe psoriasis (on Humira) and NASH, coccidioidomycosis in 2018 (on ketoconazole) presented with jaundice and abdominal distention in September 2020. She was diagnosed with cirrhosis secondary to NASH, and DILI with a MELD score of 30. Ketoconazole and Humira were held. She was medically managed with furosemide and spironolactone. Her MELD score improved to 18. In January 2021, an orthotopic liver transplant (OLTx) was attempted but due to incidental finding of extensive peritoneal mesothelioma, this was aborted. MPM was confirmed on biopsy with cytokeratin 7+, calretinin+, WT1+, and p53 reactivity. The patient was not considered a candidate for cytoreductive surgery/hyperthermic intraperitoneal chemotherapy. She had multiple psoriatic flares which were managed with prednisone. A PET scan done in June 2022 revealed mild FDG uptake in sub-centimeter bilateral axillary and pelvic LN, cirrhotic liver, and splenomegaly. The patient denied the use of alternative medicine, except for the Atkins diet. She is now being considered for OLTx Discussion: We present a unique case of spontaneous regression of MPM and its association with psoriatic flare. Previous case reports suggest a co-relation of immune-mediated response with spontaneous regression.1 The immunologically relevant cytokines IF-alpha, IF-gama, and TNF-alpha can directly inhibit the growth of mesothelioma.2 These cytokines are elevated in psoriasis, which this case highlights. Trials with immunomodulatory agents e.g. CTLA-4 inhibitor have shown promising outcomes.3 However, little information is available about regression without any chemo/radiation/biologic therapy and the role of the innate immune response leading to tumor regression.

Abstract P3128: Obesogenic Diet Promotes Endothelial-to-Mesenchymal Transition In Adipose Tissue

Vascular dysfunction and chronic inflammation are characteristics of obesity-induced adipose tissue dysfunction. Proinflammatory cytokines can drive Endothelial-to-Mesenchymal Transition (EndoMT), where endothelial cells transition into mesenchymal-like interstitial cells that leave the vascular wall and promote tissue inflammation and fibrosis. In this study, we aimed to determine whether obesogenic diet can promote EndoMT in adipose tissue. EndoMT was assessed in 1) transgenic endothelial cell lineage-tracing mice subjected to High-Fat/High-Sucrose (HF/HS) diet, 2) available single cell RNA sequencing datasets from human obese adipose tissue, and 3) cell culture models using primary human adipose-derived endothelial cells. In lineage-tracing mice, obesogenic diet induced EndoMT in a time-dependent manner at 13, 26, and 52 weeks on HF/HS diet in both subcutaneous and visceral adipose tissue. Interestingly, rate of EndoMT was greater in visceral compared to subcutaneous adipose tissue. After 52-weeks on HF/HS diet, EndoMT cells in visceral adipose tissue upregulate inflammatory response and TGF-Beta signaling transcriptional pathways compared to endothelial cells in the same adipose tissue depot. Initiation of EndoMT was also found in single cell RNA sequencing datasets of stromal cells from human obese adipose tissue, indicated by endothelial cells in these datasets containing sub-clusters with reduced Pecam1 and increased Acta2 expression. Finally, the capacity for EndoMT in primary human adipose-derived endothelial cells was validated by bulk RNA sequencing after treatment with proinflammatory cytokines (TNF-Alpha, IFN-Gamma, TGF-Beta1), confirming decreased expression of endothelial cell genes and increased expression of genes associated with EndoMT, inflammatory response, and TGF-Beta signaling. Together, these clinical and experimental findings indicate that chronic obesity can promote EndoMT in adipose endothelial cells.

The Use of Agro-Industrial Waste Rich in Omega-3 PUFA during the Weaning Stress Improves the Gut Health of Weaned Piglets

(1) Background: The weaning period is a very important stage in the pig life, as during weaning, the animals are very susceptible to pathogens and develop postweaning diarrhoea. The aim of our study was to counteract weaning stress and to improve piglets’ gut health by using a nutritional intervention consisting of a mix of agro-industrial wastes (grapeseed, flaxseed and sea blackthorn meals) rich in omega-3 PUFA. (2) Methods: Twelve cross-bred TOPIG hybrid piglets with an average body weight of 11.25 kg were randomly distributed to one of the two experimental groups: a control group fed basic corn soybean diet (control diet) and an experimental group fed a diet with a 10% mixture of grapeseed, flaxseed and sea buckthorn meals in a ratio of 3:4:1 (GFS diet). (3) Results: the GFS diet had no effect on the performance, biochemical parameters or the total antibody synthesis. GFS diet was able to significantly reduce the concentration of proinflammatory cytokines IL-1 beta and TNF-alpha and to significantly increases the expression of junction proteins (occludin, claudin 4, claudin 7 and extracellular protein matrix) at the gene or protein level as compared with control. The presence of GFS in the diet increased the abundance of Bifidobacterium and Lactobacillus species in the colonic content as well as the concentration of propionic and butyric acids. (4) Conclusions: Taken together, our results showed that agro-industrial wastes rich in omega-3 PUFA can be used as an ecological, environmentally friendly nutritional intervention for improving the negative effects associated with the weaning stress.

MECHANISM OF ACTION, EFFICACY, AND SAFETY OF PROPOLIS FOR THE MANAGEMENT OF ORAL MUCOSITIS: A SYSTEMATIC REVIEW

Oral mucositis (OM) is an oral mucosal inflammation and complication of chemotherapy and/or radiotherapy. One of the natural agents that has been widely studied as an alternative therapy for OM was Propolis. This review aims to analyze the effectiveness and safety of propolis and its mechanism of action in preventing and treating OM through clinical study in human and animal models. Articles searched using the keywords “Oral Mucositis” AND “Propolis”, conducted through PubMed, ScienceDirect, and Cochrane Library databases. The inclusion criteria were: Clinical Trial and Animal Study design; in English; full paper available; published between 2016-2021; and high range of quality and articles in line to the research topic. RoB-tools JADAD Oxford Quality Scoring System and SYRCLE’s RoB tool was used for risk of bias determination. This paper writing refers to PRISMA guidelines. In vivo studies and clinical trials have shown that propolis can reduce OM Index scores in animals or OM grades in patients. Propolis can also reduce the symptoms of inflammation in OM and almost all articles stated that there were no side effects of propolis for oral mucositis. In vivo studies showed that propolis was able to inhibit pro-inflammatory markers, hypoxia markers, MPO serum levels, and TNF-alpha cytokines, but increased the expression of pS6, pAKT, NF-B, and GSH. Propolis is effective and safe to use in patients receiving chemotherapy/radiotherapy to prevent the severity and potential for OM therapy. The mechanism of action of propolis in overcoming clinical symptoms of OM is as an anti-inflammatory, antioxidant, and helps accelerate wound healing.

Linagliptin in Combination With Metformin Ameliorates Diabetic Osteoporosis Through ModulatingBMP-2 and Sclerostin in the High-Fat Diet Fed C57BL/6 Mice.

BackgroundDiabetic osteoporosis is a poorly managed serious skeletal complication, characterized by high fracture risk, increased bone resorption, reduced bone formation, and disrupted bone architecture. There is a need to investigate drugs that can improve bone health along with managing glycemic control. DPP-4 inhibitors and metformin have proven benefits in improving bone health. Here, we investigated the effects of linagliptin, a DPP inhibitor, and metformin alone and in combination to treat diabetic osteoporosis in high-fat-fed mice.MethodsC57BL/6 mice were kept on the high-fat diet (HFD) for 22 weeks to induce diabetic osteoporosis. Linagliptin (10mg/Kg), metformin (150mg/Kg), and their combination were orally administered to the diabetic mice from the 18th-22nd week. Femur and tibial bone microarchitecture together with bone mineral density (BMD) were evaluated using µCT and histopathological changes were assessed. Further, bone turnover biomarkers namely bone morphogenetic protein-2 (BMP-2), sclerostin, tartrate-resistant acid phosphatase (TRAP), osteocalcin, alkaline phosphatase (ALP), calcium, and pro-inflammatory cytokines were assessed. Additionally, metabolic parameters including body weight, fasting blood glucose (FBG), glucose & insulin tolerance, lipids profile, and leptin were measured.ResultsHFD feeding resulted in impaired bone microarchitecture, reduced BMD, distorted bone histology, and altered bone turnover biomarkers as indicated by the significant reduction in bone ALP, BMP-2, osteocalcin, and an increase in sclerostin, TRAP, and serum calcium. Interestingly, treatment with linagliptin and its combination with metformin significantly reverted the impaired bone architecture, BMD, and positively modulated bone turnover biomarkers, while metformin alone did not exhibit any significant improvement. Further, HFD induced diabetes and metabolic abnormalities (including an increase in body weight, FBG, impaired glucose and insulin tolerance, leptin, triglycerides, cholesterol), and pro-inflammatory cytokines (TNF-alpha and IL-1β) were successfully reversed by treatment with linagliptin, metformin, and their combination.ConclusionLinagliptin and its combination with metformin successfully ameliorated diabetic osteoporosis in HFD-fed mice possibly through modulation of BMP-2 and sclerostin. The study provides the first evidence for the possible use of linagliptin and metformin combination for managing diabetic osteoporosis.

New-onset lichen planus and lichen planus flare in elderly women after COVID-19 vaccination.

Dear Editor, Owing to the widespread use of COVID-19 vaccines, the number of cutaneous vaccine-related adverse events has recently increased.1 We present two cases of lichen planus in elderly women following COVID-19 mRNA vaccination. The first patient presented with new-onset LP, whereas the second patient experienced an extensive LP flare after a 10-year remission. The first patient was an 82-year-old woman who developed an extensive pruritic rash 14 days after receiving a second dose of the Pfizer-BioNTech COVID-19 vaccine. Other possible triggers such as recent infections, medication changes, or stressful events preceding the eruption were ruled out. The patient had no significant medical history, other than well-controlled arterial hypertension. On physical examination, she had polygonal, purpuric, scaly papules in the axillary region, flexural wrists and forearms, ankles, buttocks, lower back, and abdomen (Figure 1). Mucosal lesions were not observed. Dermoscopy revealed mild desquamation and Wickham's striae. Skin biopsy demonstrated irregular epidermal hyperplasia with hypergranulosis and hyperkeratosis, vacuolar degeneration of the basal layer, and dense lymphocytic infiltrate in the superficial dermis (Figure 1). Considering the clinical presentation, histopathological analysis, and absence of an alternative trigger, we diagnosed LP triggered by the mRNA COVID-19 vaccine. Thus, we began treatment with prednisolone 20 mg daily, tapering the dose over 6 weeks, which resulted in gradual improvement of skin lesions. The second patient was a 68-year-old woman with a 2-week history of an intensely pruritic eruption that occurred 14 days after receiving a second dose of the Moderna COVID-19 (mRNA-1273) vaccine. Her medical history included arterial hypertension, hyperlipidemia, obesity, HLA-B27 spondyloarthritis, and LP, which was histologically confirmed 10 years ago and successfully treated with systemic and topical corticosteroids. During the examination, erythematous to violaceous squamous papules were discovered on the trunk, buttocks, and extremities, as well as around the ankles, flexural wrists, forearms, and axillary folds, with palmar involvement as well (Figure 2). Mucosal or nail lesions were absent. Dermoscopy revealed slight scaling and Wickham's striae, and a skin biopsy confirmed lichen planus. Therefore, we introduced 30 mg prednisolone, and tapered the dose to discontinuation over 6 weeks, which lead to the resolution of skin lesions. LP is a chronic inflammatory T-cell-mediated skin disease caused by various infections, drugs, and, in rare cases, vaccines.1-6 The most common vaccines causing LP are hepatitis B and influenza vaccines. Recently, the new-onset or reactivation of LP has been associated with COVID-19 infection and vaccination. Although most reported cases of LP following COVID vaccination were localized, we observed two cases of widespread LP.2-5 The majority of vaccine-related reactions occur in middle-aged women, implying an increased risk of such reactions in women, even in elderly ones, as observed in our cases.1, 6 Both our patients developed LP 14 days after receiving the second dose of mRNA COVID-19 vaccine, which is consistent with vaccine-related LP time onset.6 The immunopathogenesis of vaccine-induced LP likely involves Th1 cell activation with upregulation of proinflammatory cytokines IL-2, TNF-alpha, and IFN-y, which leads to apoptosis of basal keratinocytes, a hallmark of LP. However, the precise mechanism underlying LP (re)activation following vaccination remains unknown. Similarly, immune dysregulation after COVID-19 vaccination may result in the (re)activation of other immune-mediated skin diseases such as psoriasis, vitiligo, acne vulgaris, pemphigus vulgaris, and certain connective tissue diseases.2, 5 Likewise, COVID-19 pandemic has underlined the importance of dermatologists and rheumatologists in providing guidance regarding immunosuppressive and biological therapy considering their extensive experience with such treatments, as well as recognizing the immune-mediated aspects of COVID-19 infection and vaccination.7 Furthermore, as a result of a better understanding of COVID-19 disease mechanisms, novel therapeutic strategies utilizing adipose-derived mesenchymal stem cells (AD-MSCs) are being developed, and given their immuno-modulatory properties, could be employed to combat the disease.8 Finally, dermatologists should be aware of new-onset or flare-ups of LP in older women caused by COVID-19 vaccination as well as the possible (re)activation of other immune-mediated skin diseases. N.H. wrote the original manuscript. T.Z., S.P., and I.B. performed literature research and contributed to the analysis of the data. M.K. and L.P.M. performed supervision, and writing—review and editing. The authors declare no conflicts of interest. Authors declare human ethics approval was not needed for this study. The data that support the findings of this study are available from the corresponding author upon reasonable request. Written informed consent was obtained from the patients for the publication of this case report and accompanying images.