Serum Cytokine Concentrations Research Articles

A Pilot Study on Burnout in Medical Students (BuMS) over an Academic Year

Background: Physician burnout is increasingly recognized as a problem in physician well-being and may negatively affect patient care outcomes. Burnout can begin at any point of training or practice, potentially as early as the first year of medical school. Thus, there is a need to characterize possible burnout in medical students as the first step to optimizing strategies for mitigation. Traditionally, burnout has been studied using survey-based variables; however, identifying novel physiological and molecular biomarkers could allow for the expansion of screening and intervention strategies. Methods: In this pilot prospective cohort study, we followed a group of preclinical 1st and 2nd year medical students (n = 9) at the University of Florida over one academic year of medical school. We collected survey responses (Maslach Burnout Inventory [MBI], Patient Health Questionnaire-9 [PHQ-9], and Perceived Stress Scale [PSS]) and measured a panel of candidate physiological biomarkers of burnout (Inflammatory Cytokine Panel, Heart Rate Variability [HRV], and Leukocyte Telomere Length). Results: In the study participants, MBI composite scores and PHQ-9 scores showed a statistically significant increase over the course of an academic year, indicating higher levels of medical student burnout. Additionally, respondents reported a statistically significant decrease in time devoted to exercise, and we measured a significant increase in body mass index (BMI) during the academic year. PSS scores showed an upward trend which was not statistically significant. Likewise, average leukocyte telomere length trended downward, but the change was not statistically significant. There were no measured changes in the serum concentration of pro-inflammatory cytokines, and time-domain heart rate variability metrics did not differ significantly between timepoints. Conclusions: This pilot study supports the notion that burnout can begin early in medical school and is detectable via survey instruments in first-year and second-year medical students even with a small sample size. Additionally, leukocyte telomere length could potentially be a useful biomarker of burnout with supporting data, but we did not observe any statistically significant changes in inflammatory cytokines or heart rate variability. Further investigation into these potential biomarkers with larger cohort sizes is required to fully characterize their clinical utility.

Serum Cytokine Profiles in Patients with Rheumatoid Arthritis Before and After Treatment with Methotrexate.

Rheumatoid arthritis (RA) is an inflammatory autoimmune illness affecting around 1% of the population globally. Cytokines have a crucial role in the pathogenesis of RA. The objectives of the present study were to compare the serum cytokine profiles between methotrexate (MTX)-treated and MTX-naive RA patient groups, MTX-treated RA patient group and healthy controls, and MTX-naive RA patient group and healthy controls. Enzyme linked immunosorbent assay (ELISA) kits were used to quantify the serum concentrations of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, IL-17, IL-6, interferon-gamma (IFN-γ), and IL-10 in 80 RA patients (48 MTX treated and 32 MTX naive) and 80 healthy controls. For all cytokine assays, absorbance was measured at 450 nm using a microplate reader (Bio-Rad). Independent sample t-test was used to compare the serum cytokine concentrations between the study groups using SPSS version 25. MTX-treated RA patient group had significantly reduced serum levels of TNF-α (36.13 ± 17.64 versus 45.82 ± 23.07, *P = 0.037), IL-17 (307.85 ± 151.74 versus 435.42 ± 241.19, **P = 0.006), and IFN-γ (414.93 ± 212.13 versus 527.15 ± 269.61, *P = 0.041) compared to MTX-naive RA patients. Both MTX-treated and MTX-naive RA patient groups had significantly high serum levels of TNF-α, IL-1β, IL-17, IL-6, IFN-γ, and IL-10 when compared to healthy controls (***P < 0.001). Downregulation of the serum concentrations of certain key cytokines, viz. TNF-α, IL-17, and IFN-γ, demonstrates the anti-inflammatory effect of MTX in RA patients.

Safety and efficacy of remote ischemic conditioning in adult moyamoya disease patients undergoing revascularization surgery: a pilot study.

Revascularization surgery for patients with moyamoya disease (MMD) is very complicated and has a high rate of postoperative complications. This pilot study aimed to prove the safety and efficacy of remote ischemic conditioning (RIC) in adult MMD patients undergoing revascularization surgery. A total of 44 patients with MMD were enrolled in this single-center, open-label, prospective, parallel randomized study, including 22 patients assigned to the sham group and 22 patients assigned to the RIC group. The primary outcome was the incidence of major neurologic complications during the perioperative period. Secondary outcomes were the modified Rankin Scale (mRS) score at discharge, at 90 days post-operation, and at 1 year after the operation. The outcome of safety was the incidence of adverse events associated with RIC. Blood samples were obtained to monitor the serum concentrations of cytokines (VEGF, IL-6). No subjects experienced adverse events during RIC intervention, and all patients could tolerate the RIC intervention in the perioperative period. The incidence of major neurologic complications was significantly lower in the RIC group compared with the control group (18.2% vs. 54.5%, P = 0.027). The mRS score at discharge in the RIC group was also lower than the control group (0.86 ± 0.99 vs. 1.18 ± 1.22, P = 0.035). In addition, the serum IL-6 level increased significantly at 7 days after bypass surgery in the control group and the serum level of VEGF at 7 days post-operation in the RIC group. In conclusion, our study demonstrated the neuroprotective effect of RIC by reducing perioperative complications and improving cerebral blood flow in adult MMD patients undergoing revascularization surgery. Thus, RIC seems to be a potential treatment method for MMD. ClinicalTrials.gov, identifier: NCT05860946.

Association among inflammaging, body composition, physical activity, and physical function tests in physically active women.

Inflammaging is a phenomenon that has been associated with the development and progression of sarcopenia and frailty syndrome. According to the literature, on the one side, the increase in body fat is associated with a systemic pro-inflammatory status, which consequently favors inflammaging, and on the other side, the regular practice of physical exercise can mitigate the development of this scenario. Therefore, here, we aimed to evaluate the association between inflammaging and physical factors, both body and functional, in a group of physically active older women. Seventy older women (mean age 72.66 ± 6.17 years) participated in this observational cross-sectional and were separated into the eutrophic, overweight, and obese groups. It was assessed: by bioimpedance-body fat percentage (Fat%) and total (Fat kg), skeletal muscle mass (muscle), and free fat mass both in percentage (FFM%) and total (FFMkg); by the International Physical Activity Questionnaire (IPAQ)-the time of moderate-intensity physical activity per week; by physical tests-handgrip (HG), sit-up-stand-on-the-chair in 5 repetitions (Sit-up) and vertical squat jump test (SJ); in addition to the determination of serum cytokine concentration (IL-6, TNF-α, IL-10, and IL-8), and also body mass index (BMI) and calf circumference (Calf). Higher FFM% and lower body fat (both kg and %) were found in the eutrophic group than in the other groups. The eutrophic group also performed more weekly physical activity, jumped higher, and presented not only higher serum IL-6 concentration but also an increased ratio of IL-10/IL-6, IL-10/TNF-α, IL-10/IL-8 as compared to the values found in the overweight group. The obese group presented higher body fat (kg and %) and lower FFM% than the other groups and also higher serum IL-6 concentration than the overweight group. Interestingly, several significant negative and positive correlations between body composition, physical tests, and serum cytokine concentrations were found in the eutrophic and obese groups. While the eutrophic older women group showed a remarkable regulation of the systemic inflammatory status with positive associations in the physical parameters assessed, the overweight and obese groups presented impairment regulations of the inflammaging, which could be related to less weekly physical activity and higher body fat.

Effect of treatment regimen of the rheumatoid arthritis patients on the risk of coronavirus disease 2019 by modulating the inflammatory mediators.

The therapeutic profile of the patients with rheumatoid arthritis (RA) commonly consists of immunosuppressive and anti-inflammatory compounds. Here in this research, we assessed the potential effect of drug treatment in the RA patients in increasing the risk of coronavirus disease 2019 (COVID-19) infection. In this retrospective cross-sectional study, 200 subjects with RA were recruited. The treatment profile of the subjects for the past 6months was collected. The COVID-19 diagnosis was implemented based on the standard molecular tests and clinical examinations. Serum concentration of cytokines was measured using enzyme-linked immunosorbent assay (ELISA). It was detected that there was an increased risk of COVID-19 in RA subjects receiving Etanercept (OR = 3.51, 95% CI 1.19-10.30, P = 0.022). Concentrations of Interleukin (IL)-1β, Interferon (IFN)-γ, Tumor necrosis factor (TNF)-α, IL-6, IL-17A, and IL-23 were significantly higher in the RA patients with COVID-19 relative to RA cases without COVID-19. In RA/COVID-19 cases receiving Etanercept, serum levels of TNF-α, IL-1β, and IL-6 were significantly lower than RA/COVID-19 subjects without Etanercept therapy. It seems that Etanercept therapy in RA cases might increase proneness of the COVID-19 risk in these cases. The mechanism of this increased risk may stem from suppressing a protective immunity state in the RA cases.

Building Predictive Models for Schizophrenia Diagnosis with Peripheral Inflammatory Biomarkers.

Machine learning and artificial intelligence technologies are known to be a convenient tool for analyzing multi-domain data in precision psychiatry. In the case of schizophrenia, the most commonly used data sources for such purposes are neuroimaging, voice and language patterns, and mobile phone data. Data on peripheral markers can also be useful for building predictive models. Here, we have developed five predictive models for the binary classification of schizophrenia patients and healthy individuals. Data on serum concentrations of cytokines, chemokines, growth factors, and age were among 38 parameters used to build these models. The sample consisted of 217 schizophrenia patients and 90 healthy individuals. The models architecture was involved logistic regression, deep neural networks, decision trees, support vector machine, and k-nearest neighbors algorithms. It was shown that the algorithm based on a deep neural network (consisting of five layers) showed a slightly higher sensitivity (0.87 ± 0.04) and specificity (0.52 ± 0.06) than other algorithms. Combining all variables into a single classifier showed a cumulative effect that exceeded the effectiveness of individual variables, indicating the need to use multiple biomarkers to diagnose schizophrenia. Thus, the data obtained showed the promise of using data on peripheral biomarkers and machine learning methods for diagnosing schizophrenia.

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.

Better post-operative outcomes at 1-year follow-up are associated with lower levels of pre-operative synovitis and higher levels of IL-6 and VEGFA in unicompartmental knee arthroplasty patients.

Osteoarthritis (OA) is associated with inflammation, and residual inflammation may influence outcomes following knee arthroplasty. This may be more relevant for patients undergoing unicompartmental knee arthroplasty (UKA) due to larger remaining areas of native tissue. This study aimed to: (1) characterise inflammatory profiles for medial UKA patients and (2) investigate whether inflammation markers are associated with post-operative outcomes. This prospective, observational study has national ethics approval. Bloods, synovial fluid, tibial plateaus and synovium were collected from medial UKA patients in between 1 January 2021 and 31 December 2021. Cytokine and chemokine concentrations in serum and synovial fluid (SF) were measured with multiplexed assays. Disease severity of cartilage and synovium was assessed using validated histological scores. Post-operative outcomes were measured with Oxford Knee Score (OKS), Forgotten Joint Score (FJS-12) and pain scores. The study included 35 patients. SF VEGFA was negatively correlated with pre-operative pain at rest (r -0.5, p = 0.007), and FJS-12 at six-week (r 0.44, p = 0.02), six-month (r 0.61, p < 0.01) and one-year follow-up (r 0.63, p = 0.03). Serum and SF IL-6 were positively correlated with OKS at early follow-up (serum 6weeks, r 0.39, p = 0.03; 6months, r 0.48, p < 0.01; SF 6weeks, r 0.35, p = 0.04). At six weeks, increased synovitis was negatively correlated with improvements in pain at rest (r -0.41, p = 0.03) and with mobilisation (r -0.37, p = 0.047). Lower levels of synovitis and higher levels of IL-6 and VEGFA were associated with better post-operative outcomes after UKA, which could be helpful for identifying UKA patients in clinical practice. Level IV case series.

Bushen Antai recipe ameliorates immune microenvironment and maternal-fetal vascularization in STAT3-deficient abortion-prone mice

Ethnopharmacological relevanceSpontaneous abortion (SA) is an intricate disorder affecting women of reproductive age. Previous studies have confirmed the indispensable role of signal transducer and activator of transcription (STAT) 3 in normal pregnancy. Bushen Antai recipe (BAR) is a satisfactory formula commonly used in practice, based on the rationale of traditional Chinese medicine (TCM) for SA. Aim of the studyThe current study explores the potential therapeutic effects and mechanistic insights of BAR in STAT3-deficient abortion-prone mice. Materials and methodsA STAT3-deficient abortion-prone mouse model was developed using intraperitoneal injection of stattic from embryo day (ED) 5.5 to ED9.5 among pregnant females (C57BL/6). We separately administered BAR1 (5.7 g/kg), BAR2 (11.4 g/kg), progesterone (P4), or distilled water at 10 ml/kg/day from ED0.5 until ED10.5. The embryo resorption rate and placenta-uterus structure were observed on ED10.5. The systemic immune status was examined by analyzing the frequency of immunosuppressive myeloid-derived suppressor cells (MDSCs), the ratio of two macrophage (M) subtypes, and the protein expression of associated molecules. Morphological observation, immunohistochemistry, and western blotting were used to evaluate the vascularization conditions at the maternal-fetal interface. ResultsBAR1, BAR2, or P4 treatment exerted remarkable effects in alleviating embryo resorption rate and disordered placental-uterus structure in STAT3-deficient abortion-prone mice. Western blotting indicated the deficiency of phosphorylated STAT3 and two prime target molecules, PR and HIF-1α, at the maternal-fetal interface under STAT3 inhibition. Simultaneously, BAR2 treatment significantly upregulated their expression levels. The systemic immune environment was disrupted, indicated by the reduced serum cytokine concentrations, MDSCs frequency, M2/M1 ratio, and the expression of immunomodulatory factors. Nonetheless, BAR2 or P4 treatment revived the immune tolerance for semi-allogenic embryos by enhancing the immune cells and factors. Besides, the western blot and immunohistochemistry results revealed that BAR2 or P4 treatment upregulated VEGFA/FGF2 and activated ERK/AKT phosphorylation. Therefore, BAR2 or P4 facilitated vascularization at the maternal-fetal interface in STAT3-deficient abortion-prone mice. ConclusionsBAR sustained pregnancy by reviving the systemic immune environment and promoting angiogenesis at the maternal-fetal interface in STAT3-deficient abortion-prone mice.

Immunoregulatory effects of local and systemic use of melatonin in experimental thermal trauma

High incidence of thermal trauma (TT), increased risk of infectious and non-infectious short- and long-term complications, and limited effectiveness of the therapeutic approaches are the main pre-requisites for searching and pathogenetic justification of new therapies. E.g., melatonin, an endogenous homeostatic regulator with pleiotropic properties, deserves special attention. The aim of the work was to study the effect of local application (as an original dermal film) and systemic usage of melatonin upon concentrations of circulating TNF and IL-4 in the course of experimental thermal trauma.&#x0D; The rates of wound epithelization were expressed as per cent values. The plasma concentrations of IL-4, TNF were determined with automatic enzyme immunoassay analyzer Personal LAB (Italy).&#x0D; In the series with TT, the rate of wound epithelization increased from the 5th to the 20th day of experiment. When using dermal films with MT in experimental TT on days +5, +10 and +20, an increase in the rate of wound epithelization was registered. Under the conditions of intraperitoneal MT injection in experimental TT, the rate of wound epithelization increased on days +10 and +20. A correlation analysis revealed stronger interrelations observed when applying dermal films with MT in thermal trauma, rather than in series with intraperitoneal usage of MT thus suggesting faster repair processes in the first case, along with diminished area of alterations and decreased amounts of cytokines in serum.&#x0D; In experimental TT, the concentrations of TNF and IL-4 increase in blood serum on days +5, +10 and +20. On days +10 and +20, moderate positive associations were recorded between the rate of wound epithelialization and the concentration of serum cytokines. Upon intraperitoneal use of MT in experimental TT, on days +5, +10 and +20, the rate of epithelization increases, along with decrease of TNF concentration on days +10 and +20. On days +5, +10 and +20, moderate negative associations were recorded between the rate of wound epithelization and the concentration of TNF. Under the conditions of using DP with MT with TT, the rate of wound epithelization increases on days 5, 10 and 20, the concentration of TNF decreases in serum, the concentration of IL-4 decreases on day +10. On days +5, +10 and +20, the negative correlations were recorded between the rate of wound epithelization and serum concentrations of TNF and IL-4. A comparative efficiency analysis of MT-containing dermal films versus intraperitoneal use of MT in experimental thermal trauma revealed a more accelerated epithelization of the wound along with earlier decrease in TNF, a greater number and strength of relationships between the rate of the wound epithelization and concentrations of cytokines in blood serum.

Ccrl2-centred immune-related lncRNA-mRNA co-expression network revealed the local skin immune activation mechanism of moxibustion on adjuvant arthritis mice

BackgroundMoxibustion is an important external therapy of traditional medicine that operates on some acupoints on the skin and is usually used for immune-related diseases. However, whether the immune function of the skin, especially the immune-related lncRNAs, contributes to the mechanism of moxibustion remains unclear. MethodsAdjuvant arthritis (AA) was induced by injection of Complete Freund's adjuvant (CFA) into the right hind paw of mice. Moxibustion was administered on the Zusanli (ST36) acupoint for 3 weeks. The alteration of foot volume and cytokine concentration in serum was used to evaluate the anti-inflammation effect of moxibustion. CD83 expression in the local skin of ST36 was measured by immunofluorescence staining. Transcriptome RNA sequencing (RNA-seq) and lncRNA-mRNA network analysis were performed to construct a moxibustion-induced Immune-related lncRNA-mRNA co-expression network. qRT-PCR was used to validate the RNA-seq data. ResultsMoxibustion at ST36 relieved the foot swelling, decreased the TNF-α and IL-1β concentrations in serum, and obviously increased the CD83 expression at the local skin of ST36. A total of 548 differentially expressed lncRNAs and 520 linked mRNAs were screened out. The significantly and predominately enriched Go term was inflammatory and immune response, and the main pathways related to inflammatory and immune responses include Toll-like receptor, cytokine-cytokine receptor, and MAPK signaling. The immune-related lncRNA-mRNA co-expression network showed 88 lncRNAs and 36 mRNAs, and Ccrl2 is the central hub of this network. ConclusionLocal immune activation is significantly triggered by moxibustion in ST36 of AA mice. The Ccrl2-centered immune-related lncRNA-mRNA co-expression network would be a promising target for decoding the mechanism of moxibustion for immune-related diseases.

Impact of different job types on serum cytokine levels in apparently healthy individuals: A comparative study among airline pilots, construction workers, and fitness instructors

BackgroundThe impact of occupational factors on serum cytokine concentrations has not been extensively explored. In this preliminary investigation, we measured the amounts of 12 cytokines in the serum of healthy individuals, comparing three diverse professional categories (aviation pilots, building laborers, and exercise trainers) with distinct work settings and lifestyle factors. MethodsThe study sample comprised 60 men from three distinct professional fields – airline pilots, construction laborers, and fitness trainers (20 participants per category) – who were enlisted during regular outpatient occupational health appointments. Serum levels of interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-17, tumor necrosis factor (TNF)-α, interferon (IFN)-α, and IFN-γ were measured on a Luminex® platform using a specific kit. Cytokine levels were compared among the three professional groups to determine any significant differences. ResultsAmong the three occupational groups, fitness instructors demonstrated elevated IL-4 concentrations in comparison to both airline pilots and construction laborers, with no significant difference between the latter two professions. Additionally, a stepwise increase in IL-6 levels was identified, commencing with fitness instructors presenting the lowest quantities, succeeded by construction workers, and culminating with airline pilots, who displayed the most elevated concentrations. ConclusionSerum cytokine levels in healthy individuals can exhibit variations based on their occupation. Given the unfavorable cytokine profile detected in airline pilots, it is crucial for the aviation sector to tackle potential health concerns within their employees.

P13-002-23 Cytokine Serum Concentrations and the Relationship Between Cytokines and Gastrointestinal Symptoms in Patients With Gynecological Cancer Undergoing Radiotherapy

P13-002-23 Cytokine Serum Concentrations and the Relationship Between Cytokines and Gastrointestinal Symptoms in Patients With Gynecological Cancer Undergoing Radiotherapy

Inflammatory cytokines and chemokines in obese adolescents with antibody against to adenovirus 36

Obesity in adolescents has reached epidemic proportions and is associated with the inflammatory response and viral infections. The aim of this study was to understand the profile of inflammatory cytokines and chemokines associated with the inflammatory response and metabolic syndrome (MetS) in obese adolescents with positive serology for adenovirus 36 (ADV36). Thirty-six overweight, 36 obese, and 25 severe obesity adolescents aged 10 to 16 years were included in the study. The following variables were analyzed: sex, age, body mass index (BMI), blood pressure, total cholesterol and fractions, triglycerides, glucose, serum cytokine concentrations, and ADV36 antibodies. Cytokines and chemokines were quantified by cytometry and ADV36 serology was determined by enzyme-linked immunosorbent assay (ELISA). The results showed higher levels of the cytokines interleukin-1beta (IL-1β), IL-6, IL-10 and of the chemokine interferon-gamma-inducible protein 10 (IP-10) in severe obesity adolescents compared to the obese and overweight groups, as well as in the group with MetS compared to the group without this syndrome. The frequency of ADV36-positive individuals did not differ between groups. The findings revealed differences in BMI between the obese and severe obesity groups versus the overweight group in the presence of positivity for ADV36, suggesting an association with weight gain and possibly MetS installation.

Cytokine profile in patients with vibration disease, aggravated by hypertension and obesity

Comorbid diseases which include arterial hypertension (AH) accompanied by overweight, and vibration disease (VD) are known to contribute to the mutual aggravation of occupational and cardiovascular pathology. Despite numerous studies indicating that some changes of immune system are observed when exposed to vibration, the contribution of cytokines to the comorbid course of vibration disease has not been sufficiently studied. The aim of the work is to evaluate the role of inflammatory mediators in development of hypertension and obesity in vibration disease.The cytokine profile parameters were determined by ELISA immunoassay using “Vector-Best” test systems. Some unidirectional changes in serum cytokine concentrations were found in patients with VD complicatedby AH, and those with VD without AH, being characterized by increase in pro-inflammatory IL-1β, TNFα, IL-17, anti-inflammatory IL-4 and IFNγ, and a decrease in multifunctional IL-2 cytokinem, relative to the comparison group. The groups with versus without AH differed in compensatory increase of anti-inflammatory IL-10 in the latter group. At the same time, all patients with combined VD and AH had excessive body weight, with grade 1 obesity in half of the cases, thus being significantly different from those with VD without AH. It is shown that IL-1β, IL-2, IL-4, IL-8, TNFα play an apparent role in the development of obesity. The greatest contribution is observed for IL-1β, as evidenced by the quantitative relationship of body mass index with its concentrations as assessed by logistic regression analysis. The limitation of this study may be a small sample size.It was found that the unidirectional changes in cytokines when compared to the comparison group were revealed in patients with VD complicated by AH, and in patients with VD without AH. In the subjects with VD and AH, these changes were more pronounced, which may suggest the hypertension is an aggravating factor. The established relationship between individual cytokines (IL-1β, IL-2, IL-4, IL-8, TNFα) and BMI indicates their participation in development of obesity. All these findings may provide a basis for development of preventive and therapeutic measures that reduce the risk of hypertension and obesity in the people working under permanent exposure of vibration.

Network pharmacology-based approach to explore the underlying mechanism of sinomenine on sepsis-induced myocardial injury in rats.

Background: Sepsis, a systemic disease, usually induces myocardial injury (MI), and sepsis-induced MI has become a significant contributor to sepsis-related deaths in the intensive care unit. The objective of this study is to investigate the role of sinomenine (SIN) on sepsis-induced MI and clarify the underlying mechanism based on the techniques of network pharmacology. Methods: Cecum ligation and puncture (CLP) was adopted to induce sepsis in male Sprague-Dawley (SD) rats. Serum indicators, echocardiographic cardiac parameters, and hematoxylin and eosin (H&E) staining were conducted to gauge the severity of cardiac damage. The candidate targets and potential mechanism of SIN against sepsis-induced MI were analyzed via network pharmacology. Enzyme-linked immunosorbent assay was performed for detecting the serum concentration of inflammatory cytokines. Western blot was applied for evaluating the levels of protein expression. Terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end labeling assay was applied to assess cardiomyocyte apoptosis. Results: SIN significantly improved the cardiac functions, and attenuated myocardial structural damage of rats as compared with the CLP group. In total, 178 targets of SIN and 945 sepsis-related genes were identified, and 33 overlapped targets were considered as candidate targets of SIN against sepsis. Enrichment analysis results demonstrated that these putative targets were significantly associated with the Interleukin 17 (IL-17) signal pathway, inflammatory response, cytokines-mediated signal pathway, and Janus Kinase-Signal Transducers and Activators of Transcription (JAK-STAT) pathway. Molecular docking suggested that SIN had favorable binding affinities with Mitogen-Activated Protein Kinase 8 (MAPK8), Janus Kinase 1 (JAK1), Janus Kinase 2 (JAK2), Signal Transducer and Activator of Transcription 3 (STAT3), and nuclear factor kappa-B (NF-κB). SIN significantly reduced the serum concentration of Tumor Necrosis Factor-α (TNF-α), Interleukin 1 Beta (IL-1β), Interleukin 6 (IL-6), Interferon gamma (IFN-γ), and C-X-C Motif Chemokine Ligand 8 (CXCL8), lowered the protein expression of phosphorylated c-Jun N-terminal kinase 1 (JNK1), JAK1, JAK2, STAT3, NF-κB, and decreased the proportion of cleaved-caspase3/caspase3. In addition, SIN also significantly inhibited the apoptosis of cardiomyocytes as compared with the CLP group. Conclusion: Based on network pharmacology analysis and corresponding experiments, it was concluded that SIN could mediate related targets and pathways to protect against sepsis-induced MI.

Effect of topical drugs on the local concentrations of IL-5 and mometasone furoate in chronic polypous rhinosinusitis

Long-term persistence of inflammation is the main factor of pathogenesis in chronic polypous rhinosinusitis (CRSwNP). Prospectives of current clinical otorhinolaryngology include modification of drugs containing topical glucocorticosteroids in order to increase local bioavailability, reduce the concentration of glucocorticosteroids, and alleviate the local inflammatory response. The aim of our work was to evaluate the effect of urea, magnesium sulfate, and mannitol on the concentrations of mometasone furoate and IL-5 in the polypous tissue of patients with CRSwNP. The study included 146 patients with CRSwNP aged 18 to 62 years. Biopsies of polypous tissue were taken during polypotomy. In the fluid samples obtained by microdialysis of polypous tissues, mometasone concentrations were determined by high-performance liquid chromatography. Concentrations of IL-3, IL-4, IL-5 and IL-10 cytokines in blood serum, and IL-5 in microdialysates of polypous tissue were measured by ELISA technique. The average age of patients with CRSwNP was 42.9±7.2 years, with prevalence of male patients (n = 93, 63.7%) over females (n = 53, 36.3%). Severity parameters of the disease by the SNOT-22 scale were significantly higher in CRSwNP patients than in control group (p ≤ 0.05). The results of biochemical analysis of microdialysates from polypous tissue showed increased content of Na+ ions in the resistant clinical cases of CRSwNP compared with the sensitive and dependent clinical forms (p &lt; 0.05). Curves with the level of mometasone furoate were obtained in all the studied samples. It was found that the addition of magnesium sulfate solution to mometasone increases the Cmax of mometasone to 154 ng/mL in biological fluids after microdialysis. When urea solution is added to mometasone, Cmax increases to 198 ng/mL. In the native nasal polypous tissue dialysate, IL-5 levels were 89±2.01 pg/mL. Two hours after the addition of mometasone furoate, IL-5 concentrations decreased to 61±3.5 pg/mL, upon addition of urea, it changed to to 69±2.98 pg/mL (p ≤ 0.01, compared with the control without the addition of drugs). Analysis of serum cytokine levels showed that IL-5 concentrations in the patients with CRSwNP significantly exceeded (&gt; 3-fold) those in the control group (p ≤ 0.05). There was a trend for increase of IL-3 and IL-4 (2-fold) and to decreased IL-10 levels in blood sera (&gt; 1.5-fold) in patients with CRSwNP compared with controls. These results may be helpful for development of novel clinical approaches, in order to increase bioavailability of topically administered glucocorticosteroids, thus allowing to promote the effectiveness of basic therapy of CRSwNP and to develop therapy for the prevention of resistant forms of polyposis.

Elevated GRO-α and IL-18 in serum and brain implicate the NLRP3 inflammasome in frontotemporal dementia

Neuroinflammation is a hallmark of frontotemporal dementia (FTD), a heterogeneous group of proteinopathies characterized by the progressive degeneration of the frontal and temporal lobes. It is marked by microglial activation and subsequent cytokine release. Although cytokine levels in FTD brain and CSF have been examined, the number of cytokines measured in each study is limited and knowledge on cytokine concentrations in FTD serum is scarce. Here, we assessed 48 cytokines in FTD serum and brain. The aim was to determine common cytokine dysregulation pathways in serum and brain in FTD. Blood samples and brain tissue samples from the superior frontal cortex (SFC) were collected from individuals diagnosed with behavioral variant FTD (bvFTD) and healthy controls, and 48 cytokines were measured using a multiplex immunological assay. The data were evaluated by principal component factor analysis to determine the contribution from different components of the variance in the cohort. Levels of a number of cytokines were altered in serum and SFC in bvFTD compared to controls, with increases in GRO-α and IL-18 in both serum and SFC. These changes could be associated with NLRP3 inflammasome activation or the NFκB pathway, which activates NLRP3. The results suggest the possible importance of the NLRP3 inflammasome in FTD. An improved understanding of the role of inflammasomes in FTD could provide valuable insights into the pathogenesis, diagnosis and treatment of FTD.

ОЦЕНКА ЗНАЧИМОСТИ ВЗАИМОСВЯЗИ МЕЖДУ ПОКАЗАТЕЛЯМИ РЕМОДЕЛИРОВАНИЯ СОСУДОВ, ВОСПАЛИТЕЛЬНОГО ОТВЕТА И ПОЛИМОРФИЗМА ГЕНОВ ДИСФУНКЦИИ ЭНДОТЕЛИЯ У ПАЦИЕНТОВ МУЖСКОГО ПОЛА С ИНФАРКТОМ МИОКАРДА В ЗАВИСИМОСТИ ОТ НАЛИЧИЯ ИЛИ ОТСУТСТВИЯ ГИПЕРТОНИЧЕСКОГО АНАМНЕЗА

Introduction. Endothelial dysfunction, arterial hypertension and surrogate events of myocardial infarction are interrelated phenomena, while the genetic predisposition in these patients is an independent risk factor for the development of cardiovascular pathology. Aim. The aim of the study was to evaluate the significance of the interaction of associations of polymorphic loci of adhesion molecule genes (immunoglobulin VCAM1), MCP-1 chemokine, vascular endothelial growth factor (VEGFA), nitric oxide control enzyme (DDAH1), markers of the inflammatory response, vascular remodeling and endothelial dysfunction in patients with male myocardial infarction, depending on the presence or absence of arterial hypertension in history in the development of myocardial infarction. Material and methods. 667 male patients and 621 practically healthy individuals in the comparison group were examined. Patients underwent determination of general clinical parameters, serum concentrations of monocyte-chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGFA), cytokines (IL-1,6; TNF-α), molecular genetic study, measurement of media intima thickness of the common carotid artery. Selected in the group of patients with myocardial infarction, individuals with or without a history of arterial hypertension. Results and discussion. As a result of a comparative analysis of groups of patients with and without a history of arterial hypertension with a group of healthy individuals, significant differences in the frequencies of genotypes and alleles for the following polymorphic gene loci were revealed: MCP-1, VEGFA, DDAH1, VCAM1. The serum concentration of cytokines (IL-1,6; TNF-α) in patients, regardless of the presence or absence of a history of hypertension, significantly exceeded the level in the comparison group. The VEGFA index in patients with myocardial infarction with arterial hypertension was significantly increased compared to the serum level in patients with myocardial infarction without arterial hypertension. Conclusion. A higher significance of the interaction between indicators of vascular remodeling, inflammation, endothelial dysfunction gene polymorphism in the development of myocardial infarction in patients with a history of hypertension, in contrast to non-hypertensive patients with myocardial infarction, was revealed, which indicates a powerful contribution of arterial hypertension to the etiology and implementation of myocardial infarction.

Peripheral blood immune cell and cytokine profiling of patients receiving corticosteroids for immune checkpoint inhibitor-related adverse events: Steroid dose and duration.

e14694 Background: Immune checkpoint inhibitors (ICI) have dramatically extended survival in numerous malignancies, but many patients treated with ICIs experience severe immune-related adverse events (irAEs) that can affect virtually any organ. Corticosteroids (steroids) tapered over weeks to months are usually effective but may affect ICI anti-tumor activity. Tapers are often imprecise, resulting in over- or undertreatment of some patients. There are no known blood biomarkers to guide steroid taper management. We therefore profiled peripheral blood immune cells and cytokines at key timepoints to identify biomarkers of response to steroids for irAE treatment. Methods: We collected blood samples before ICI, during irAE, at three steroid dose levels (high, intermediate, and low), and post-steroids from patients treated with ICI therapy at Yale Cancer Center. We measured 48 serum cytokines with a multiplexed immunoassay and immunophenotyped 40 markers on peripheral blood mononuclear cells with mass cytometry (CyTOF). We correlated changes in serum cytokine concentration across timepoints with clinical features. We applied a cluster analysis to the CyTOF data to identify immune cell types and their change in frequency across timepoints. Results: We evaluated 12 patients treated with anti-PD-1 + anti-CTLA-4 therapy who experienced at least 1 severe irAE (11 patients with melanoma, 1 patient with renal cell carcinoma). Five patients had colitis, 6 had hepatitis, and 1 had hepatitis and enteritis. Concentrations of TNFα, IFNγ-mediated cytokines (IL-12p40, IL-18, IL-27), and T cell homing chemokines (CXCL9, CXCL10) increased during irAEs and decreased during steroid therapy without significant change across steroid dose levels (p &lt; 0.05). We observed the largest changes in patients who experienced another irAE within 3 months (n = 5). IL-6 was notably higher in patients who required steroid taper escalation for recurrent symptoms before or soon after completing steroid therapy (n = 8, p = 0.02). Compared to other T cell subtypes, the frequency of CD4 T effector memory (Tem) cells increased during irAE and decreased upon completing steroid therapy but did not return to baseline (p &lt; 0.01). Conclusions: In patients treated with anti-CTLA-4 + anti-PD-1 therapy, IL-6 and IFNγ-mediated cytokines were elevated during irAEs and suppressed at multiple steroid dose levels despite symptom recurrence during the steroid taper. Increased CD4 Tem cell frequency during irAEs may reflect circulating pathogenic T cells, and the delayed decrease and incomplete return to baseline could be due to the severity of irAEs represented in this study. Specific cytokines may predict the occurrence of multiple irAEs and the need for a longer steroid taper. Additional longitudinal analyses of steroid effects on irAEs are needed to validate these findings.