Serum Concentrations Of TNF-α Research Articles

Comparison of serum calprotectin, a marker of neutrophil activation, and other mediators of inflammation in response to alcohol consumption.

Previous studies have indicated that heavy alcohol intake stimulates inflammation and impairs the body's ability to regulate inflammation. The aim of this study was to compare changes in neutrophil calprotectin and a wide spectrum of other inflammatory mediators in response to heavy alcohol drinking. Serum calprotectin (a marker of neutrophil activation), suPAR, CD163, and pro- (IL-6, IL-8, TNF-α) and anti-inflammatory (IL-10, TGF-β) cytokines were measured from 61 alcohol-dependent subjects (46 men, 15 women, mean age 43.6±11.0 years) at the time of admission for detoxification and after 8±2 days of abstinence. These biomarkers were also measured from age- and sex-matched healthy controls representing abstainers or light drinkers. The clinical assessments included detailed clinical interviews on the amounts and patterns of alcohol consumption and assays for biomarkers of alcohol consumption (GGT, CDT, MCV, GGT-CDT) and liver function (AST, ALT). The subjects with alcohol use disorder showed significantly higher concentrations of serum calprotectin (p<0.0005), suPAR (p<0.01), CD163 (p<0.01), IL-6 (p<0.0005), IL-8 (p<0.0005), TNF-α (p<0.001), and IL-10 (p<0.0005) than healthy controls. These inflammatory mediators, except for CD163, remained elevated after the 8 ± 2-day period of supervised abstinence, which resulted in significant decreases in the biomarkers of alcohol consumption and indices of liver status. The AUC (0.855) for serum calprotectin in differentiating between the heavy drinkers and healthy controls was equal or equivalent with those of the conventional biomarkers of alcohol consumption (GGT:0.835 or CDT:0.803). The data indicate that neutrophil calprotectin is released in response to heavy alcohol intake in a sensitive manner and may be associated with perpetuation of inflammation in patients with alcohol use disorder. Serum calprotectin may also prove to be a useful biomarker for inflammatory activity in alcohol-consuming patients.

Improvement in recurring nivolumab-induced pneumonitis with repetitive administration of infliximab in a patient with head and neck cancer: A case report

Severe pneumonitis induced by nivolumab, an anti-programmed cell death-1 monoclonal antibody, is a rare but potentially fatal immune-related adverse event. In cases of steroid-refractory pneumonitis, an appropriate therapeutic strategy using anti-tumor necrosis factor-α (TNF-α) antibody has not been established. A 59-year-old female was diagnosed with hypopharyngeal squamous cell carcinoma. Previous therapies including chemoradiotherapy and throat laryngectomy were performed, but metastatic recurrence appeared in the intrapulmonary and mediastinal lymph nodes. The patient was administered nivolumab. On the 14th day of nivolumab administration, the patient experienced dyspnea and computed tomography of the chest showed multiple consolidations in the right lung. She was diagnosed with nivolumab-induced pneumonitis. Because the pneumonitis was refractory to steroid therapy, she was administered infliximab, and the pneumonitis improved. On the 72nd and 101st days of nivolumab administration, nivolumab-induced pneumonitis re-appeared with an elevated serum TNF-α concentration. In each occurrence of pneumonitis, repetitive administration of infliximab improved the pneumonitis. Repetitive administration of infliximab may be effective for treating recurrent nivolumab-induced pneumonitis that is associated with an increased serum TNF-α concentration.

Noni (Morinda citrifolia L.) Fruit Polysaccharides Regulated IBD Mice Via Targeting Gut Microbiota: Association of JNK/ERK/NF-κB Signaling Pathways.

Inflammatory bowel disease (IBD) is a disease characterized by intestinal inflammation with immune dysregulation and intestinal microecological imbalance. In a dextran sulfate sodium salt (DSS)-induced IBD mouse model, noni (Morinda citrifolia L.) fruit polysaccharides (NFP) with homogalacturonan and rhamnogalacturonan-I domain decreased the concentration of serum LPS, TNF-α, and IL-17 by 84, 42, and 65%, respectively. It was abolished when intestinal microbiota were depleted by antibiotics. Sequencing analysis of gut microbiota showed an attenuated disruption of the microbial composition in the DSS+NFP group. Targeted metabolomic analysis revealed that NFP upregulated the content of acetic acid, propionic acid, and butyric acid by onefold but reduced isobutyric acid and isovaleric acid contents. NFP also inhibited JNK, ERK, and NF-κB phosphorylation of IBD mice. Taken together, the mechanism of NFP alleviating IBD is related to the intestinal microecological balance to inhibit inflammatory signaling pathways. This study provides a basis for NFP as a cheap intervention for the prevention and treatment of IBD patients.

Lever positioning manipulation combined with pulsed electric field on the analgesic effect of patients with lumbar disc herniation and its influence on IL-1β and TNF-α

To observe the analgesic effect of lever positioning manipulation combined with pulsed electric field on patients with lumbar disc herniation and the influence on serum IL-1β and TNF-α. From January 2018 to March 2019, 58 patients with lumbar disc herniation were included in the study, which were randomly divided into observation group and control group by digital table method. Observation group of 29 cases, including 16 males and 13 females, aged (38.03±11.29) years old, were treated with lever positioning manipulation combined with pulsed electric field. The 29 cases in control group, including 17 males and 12 females, aged (38.21±9.16) years old, were treated with pulsed electric field. Both groups of patients were treated 3 times a week, once every other day, 3 times as a course of treatment. After 2 courses of treatment, the two groups of patients were scored before and after treatment by the numeric rating scales (NRS);at the same time, the serum levels of IL-1β and TNF-α were measured before and after treatment. The NRS scores of observation group and control group were 4.21±1.76, 4.66±1.61 before treatment, and 1.28±0.84, 2.10±1.35 after treatment, respectively. The NRS scores of the observation group after treatment was significantly lower than that of the control group (P<0.05). After treatment, the concentrations of IL-1β and TNF-α in both groups became lower(P<0.05). The levels of IL-1β in observation group and control group before treatment were (119.01±69.65), (112.23±78.43) pg /ml, and after treatment were (59.78±36.60), (77.51±40.46) pg/ml, respectively. The levels of TNF-α in observation group and control group before treatment were (1.68± 1.13), (1.74±0.70) pg /ml, and after treatment were (1.14±0.56), (1.45±0.58) pg /ml, respectively. The change of IL-1β and TNF-α in observation group was better than that in control group (P<0.05). The lever positioning manipulation combined with pulsed electric field has a good analgesic effect on patients with lumbar disc herniation, and it has a significant impact on the patient's serum IL-1β and TNF-α concentration, which can be used as a clinical guide. However, the synergistic effect of lever positioning technique combined with pulsed electric field and guidelines for clinical treatment need further research.

Serum IL-6 and TNF-α Levels Are Correlated with Disease Severity in Patients with Ankylosing Spondylitis.

Previous studies have shown that a number of cytokines participate in the regulation of ankylosing spondylitis (AS). To investigate the potential role of interleukin (IL)-6 and tumor necrosis factor- α (TNF-α) in AS pathogenesis, this study examined the serum levels of IL-6 and TNF-α in patients with AS and its clinical association with disease activity. The serum concentrations of IL-6 and TNF-α from 80 patients with AS and 46 healthy control patients (HCs) were examined by electrochemiluminescence immunoassay. The correlations between the serum IL-6 and TNF-α levels and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), computed tomography (CT) imaging-based classification, and laboratory indicators were analyzed using the Spearman correlation test. Compared to HCs, patients with AS showed higher levels of IL-6 and TNF-α. There was also a positive correlation between the serum IL-6 and TNF-α levels and the BASDAI, the progression of AS, and the CT imaging-based classification. The serum levels of IL-6 correlated closely with C-reactive protein and the erythrocyte sedimentation rate. More important, patients with AS with hip joint involvement exhibited a significant elevation of serum levels of TNF-α, and higher IL-6 was detected in patients with the involvement of joints other than the hip and sacroiliac joints. The serum levels of IL-6 and TNF-α can function as important indicators for auxiliary diagnosis and disease activity evaluation of AS.

Inhibitory Effects of Palmatine on P2X7 Receptor Expression in Trigeminal Ganglion and Facial Pain in Trigeminal Neuralgia Rats

Trigeminal Neuralgia (TN) refers to recurrent severe paroxysmal pain in the distribution area of the trigeminal nerve, which seriously affects the quality of life of patients. This research applied the chronic constriction injury of the infraorbital nerve (CCI—ION) approach to induce an animal model of TN in rats. The mechanical pain threshold of each group of rats was determined postoperatively; the expression of P2X7 receptor in trigeminal ganglion (TG) was assessed by qRT-PCR, immunofluorescence and Western blot; and the changes of the proinflammatory cytokines IL-1β and TNF-α in serum of rats were detected by ELISA. The results showed that the administration of palmatine in the TN rats could reduce the mechanical pain threshold, significantly decrease the expression of P2X7 receptor in TG, and lower the serum concentrations of IL-1β and TNF-α, compared to the sham group. In addition, the phosphorylation level of p38 in TG of TN rats was significantly decreased after treatment with palmatine. Likewise, inhibition of P2X7 expression by shRNA treatment could effectively counteract the adversary changes of pain sensitivity, IL-1β and TNF-α production, and p38 phosphorylation in TN rats. Our data suggest that palmatine may alleviate mechanical facial pain in TN rats possibly by reducing the expression of P2X7 receptor in TG of TN rats, which may be attributable to inhibiting p38 phosphorylation and reducing the release of IL-1β and TNF-α.

Acute effect of two exercise intensity programs on interleukin-6, interleukin-1 beta and tumour necrosis factor-α in female futsalists

Duration, intensity, and type of exercise can affect serum cytokine levels and change inflammatory indices. The present study aimed to examine the acute effect of two different exercise intensity programs on levels of circulating interleukin (IL)-6, IL-1β and tumour necrosis factor-α (TNF-α) in athletes. Eleven female futsal players aged 20.6±1.2 years completed this cross-over study. Participants performed, either a MI (moderate intensity: 60-65%) or a HI (high intensity: 75-80% heart rate reserve) exercise program. The study was performed on different days separated by a 1-week washout period. Each session consisted of 30 min running, either MI or HI. Blood samples were taken before (Pre) and immediately after (Post) each exercise session from an antecubital vein by venous puncture in a seated position. A Student’s t-test (P&lt;0.05) was used to examine any difference between Pre and Post values. The results showed that IL-6 (P=0.22), IL-1β (P=0.90) and TNF-α (P=0.63) serum concentrations were not significantly different after moderate-intensity exercise. Similarly, high-intensity exercise did not significantly change serum concentrations of TNF-α (P=0.63), and IL-1β (P=0.18). However, HI caused a significant increase in IL-6 (P=0.04). A significant correlation was observed only between IL-1β and IL-6 (r=-0.761, P=0.01) after MI exercise. Based on the findings of the present study, the intensity of exercise can affect some cytokines, such as IL-6 in female futsal players.

Effects of Folic Acid Supplementation on Inflammatory Markers: A Grade-Assessed Systematic Review and Dose-Response Meta-Analysis of Randomized Controlled Trials.

: BackgroundIt has been theorized that folic acid supplementation improves inflammation. However, its proven effects on inflammatory markers are unclear as clinical studies on this topic have produced inconsistent results. To bridge this knowledge gap, this systematic review and meta-analysis of randomized controlled trials (RCTs) aimed to evaluate the effects of folic acid supplementation on serum concentrations of the inflammatory markers C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Methods: To identify eligible RCTs, a systematic search up to April 2021 was completed in PubMed/Medline, Scopus, Web of Science, EMBASE, Cochrane databases, and Google Scholar using relevant keywords. A fix or random-effects model was utilized to estimate the weighted mean difference (WMD) and 95% confidence interval (95% CI). Results: Twelve RCTs were included in the present meta-analysis. The pooled analysis revealed that serum concentrations of CRP (WMD: −0.59 mg/L, 95% CI −0.85 to −0.33, p < 0.001) were significantly reduced following folic acid supplementation compared to placebo, but did not affect serum concentrations of IL-6 (WMD: −0.12, 95% CI −0.95 to 0.72 pg/mL, p = 0.780) or TNF-α (WMD: −0.18, 95% CI −0.86 to 0.49 pg/mL, p = 0.594). The dose–response analysis demonstrated a significant relationship between an elevated dosage of folic acid supplementation and lower CRP concentrations (p = 0.002). Conclusions: We found that folic acid supplementation may improve inflammation by attenuating serum concentrations of CRP but without significant effects on IL-6 and TNF-α. Future RCTs including a larger number of participants and more diverse populations are needed to confirm and expand our findings.

Targeting T-Cell Receptor Signaling Pathway in Angioimmunoblastic T-Cell Lymphoma

[Backgrounds] Angioimmunoblastic T-cell lymphoma (AITL) belongs to nodal T-cell lymphoma with T follicular helper (TFH) phenotype. Both the G17V RHOA mutation and the loss-of-function TET2 mutation have been identified in 70% of AITLs. We previously demonstrated that G17V mutant RHOA augmented T-cell receptor (TCR) signaling though binding to VAV1, an essential component of the T-cell signaling pathway (Leukemia, in press). However, it remains elusive whether inhibition of TCR pathway serves as an effective treatment in AITL. [Methods] Activation of VAV1 and PLCgamma1 in response to TCR stimulation was examined in Jurkat cells inducibly expressing wild-type and G17V mutant RHOA under the treatment with dasatinib, a multi-kinase inhibitor. Nuclear factor of activated T cell (NFAT) activity was similarly examined in Jurkat cells transiently transfected with wild-type and G17V mutant RHOA. Mice expressing G17V RHOA under CD2 promoter (G17VRHOA mice) were crossed with MxCre Tet2f/f mice and pIpC was given to the offspring 4 weeks after birth to obtain Tet2-/- and Tet2-/-G17VRHOA mice. Whole transcriptome analysis was performed for CD4+ T cells purified from the spleen of tumor developed mice. Serum cytokines were examined by BD Cytometric Beads Assay. Cells derived from tumors developed in Tet2-/-G17VRHOA mice were transplanted into nude mice. Dasatinib was orally given at 5 mg/kg/day for 14 days. [Results] The G17V mutant RHOA enhanced phosphorylation of VAV1 and PLCgamma1, and NFAT reporter activity in Jurkat cells. The phosphorylation and the NFAT activity were efficiently blocked by dasatinib at 10 nM. Approximately 70% of Tet2-/-G17VRHOA mice died around 40 wo, while 14% of Tet2-/- and none of G17VRHOA mice died at the same age (p<0.05). All the moribund Tet2-/-G17VRHOA mice presented splenomegaly and lymphadenopathy. The lymph nodes and the spleens showed diffuse infiltration of tumor cells expressing TFH markers, i.e., CD4, PD1, and ICOS. The aberrant increase of high endothelial venules was observed in the swollen lymph nodes, mimicking human AITL. Whole transcriptome analysis followed by gene-set enrichment analysis revealed enrichment of Il-2/STAT5, Il-6/JAK-STAT3, and TNFα/NFκB signaling pathways in Tet2-/-G17VRHOA CD4+ spleen cells. The serum concentrations of Il-2, Il6, and TNFα in Tet2-/-G17VRHOA were significantly higher than control at the same age (p<0.05). The CD4+ cells in the enlarged spleen and the swollen lymph nodes in Tet2-/-G17VRHOA mice at 40 wo showed oligoclonal rearrangement of TCR genes. Nude mice transplanted with tumor-derived cells showed splenomegaly and lymphadenopathy preceded by elevation ofthe serum concentrations of Il-2, Il-6 and TNFα (before vs. after the transplantation, 4.33±0.14 vs. 4.80±0.37, 5.85±2.96 vs. 34.92±15.49, and 7.68±2.08 vs. 45.70±33.72, p<0.01, respectively). Mice given with dasatinib starting at 10 days after the transplantation showed higher overall survival compared to the vehicle control (p<0.05, Figure 1). The serum concentrations of Il-2, Il-6, and TNFα were decreased in dasatinib group (before vs. after the dasatinib treatment, 4.85±0.40 vs. 4.28±0.18, 38.23±18.74 vs. 10.01±7.38, and 48.37±34.6 vs. 18.67±12.37, p<0.05, respectively).Discussions and Conclusion: The combination of G17V mutantRHOAexpression and Tet2 deletion led to development of AITL-like tumors in mice, which were hughly effectively treated by dasatinib. This model may also be useful to examine novel treatment strategies for AITL, other than dasatinib. [Display omitted] DisclosuresChiba:Nippon Shinyaku: Honoraria, Research Funding.

The Effect of Oral Magnesium Supplementation on Inflammatory Biomarkers in Adults: A Comprehensive Systematic Review and Dose-response Meta-analysis of Randomized Clinical Trials.

This is a comprehensive systematic review and dose-response meta-analysis evaluating the effects of oral magnesium supplementation on inflammatory biomarkers including C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) among adults. The major databases including PubMed, Web of Science, Embase, and Google Scholar were searched for relevant publications until December 14, 2020, using appropriate keywords. The Cochrane Collaboration tool was used to assess the quality of each study. We also performed a subgroup analysis to identify probable sources of heterogeneity. A total of 18 studies with 927 participants fulfilled the eligibility criteria and included in this meta-analysis. Our results indicate that the supplementation with magnesium had no statistically significant effect on serum concentrations of CRP (WMD, - 0.49; 95% CI, - 1.72 to 0.75mg/L; P = 0.44), IL-6 (WMD, - 0.03; 95% CI, - 0.40 to 0.33pg/mL; P = 0.86), and TNF-α (WMD, 0.12; 95% CI, - 0.08 to 0.31pg/mL; P = 0.24) compared with controls. In addition, based on dose-response assessment, no significant non-linear association was found between magnesium supplementation dosage or duration on serum CRP and IL-6 concentrations. The findings of the present systematic review and meta-analysis did not support the notion that oral magnesium supplementation could have favorable effects on CRP, IL-6, and TNF-α in the adult population.

Metabolic shift precedes the resolution of inflammation in a cohort of patients undergoing bariatric and metabolic surgery

Bariatric and metabolic surgery has shown to promote weight loss and reduce systemic inflammation. However, the sequence and timing of events regarding metabolic improvement and inflammation resolution has been rarely explored. Furthermore, data on inflammatory markers of Th17 and Th1 cell responses after bariatric surgery is scarce. We conducted a prospective study in subjects with obesity that underwent bariatric and metabolic surgery, with follow-ups at 3 and 6 months. Anthropometric and metabolic markers such as insulin levels, HOMA-IR, and lipid parameters declined significantly 3 months after surgery; while hs-CRP, TNF-α, IL-1β, IL-6, and IL-8 serum concentrations decreased 6 months after the procedure. Concentrations of Th1 signature and driver cytokines, particularly IFN-γ, IL-12, and IL-18, and of Th17 driver IL-23 also decreased significantly after 6 months. Significant positive correlations between triglyceride levels and hs-CRP, IL-1β, and IFN-γ concentrations, and between Apo B and IFN-γ levels were observed 6 months after bariatric and metabolic surgery. In addition, BMI was associated with hs-CRP and TNF-α concentrations. Fat mass correlated with hs-CRP, TNF-α, and IL-12. Analysis of the temporality of metabolic and inflammatory events suggests that improvement in the metabolic status occurs before resolution of systemic inflammation and may be a requisite for the later event.

Scutellarin ameliorates colitis-associated colorectal cancer by suppressing Wnt/β-catenin signaling cascade

Dysregulated Wnt/β-catenin signaling pathway plays a critical role in the pathogenesis of colorectal cancer (CRC). Scutellarin, a flavonoid compound in Scutellaria barbata, has been reported to suppress CRC, with the action mechanism elusive. In this study, Scutellarin was found to inhibit the carcinogenesis of colitis-associated cancer (CAC) in mice caused by azoxymethane/dextran sulfate sodium, with alleviation of pathologic symptoms. Besides, Scutellarin attenuated mouse serum concentrations of TNF-α and IL-6, heightened Bax expression and diminished B-cell lymphoma-2 (Bcl-2) level in CAC tissues of mice, through down-regulating Wnt/β-catenin signaling cascade. In CRC HT-29 cells, Scutellarin retarded the proliferation and migration, induced apoptosis, with boosted Bax expression and decreased Bcl-2 level, which may be attributed to its repression of Wnt/β-catenin signals in HT-29 cells. Our findings demonstrate that Scutellarin may ameliorate colitis-associated colorectal cancer by weakening Wnt/β-catenin signaling cascade.

Effect of houttuynia on improving lung injury in chronic obstructive pulmonary disease by regulating the TLR4 signaling pathway.

This study aimed to investigate the effects and mechanisms of houttuynia on lung pathological injury in rats with chronic obstructive pulmonary disease (COPD). Rats were randomly divided into the normal control, COPD model (model), low‐dose treatment (low, 5 mg/kg), middle‐dose treatment (middle, 10 mg/kg), and high‐dose treatment (high, 25 mg/kg) groups. The COPD rat model was induced by smoking combined with intratracheal instillation of lipopolysaccharide. The treatment groups were given Houttuynia by gavage at 30 min before smoking. The IL‐6, IL‐1β, and TNF‐α concentrations in serum and BALF were determined by ELISA. The pathological morphology was detected by HE staining. The apoptosis cell number was evaluated by TUNEL assay. Apoptotic proteins (caspase‐3 and caspase‐9) were measured by IHC assay in lung tissues. The relative proteins [TLR4, MyD88, and p‐NF‐κB(p65)] were evaluated by Western blot assay in lung tissues. Compared with the model group, the low, middle, and high groups could reduce pulmonary congestion, edema, inflammatory cell infiltration, and apoptosis cell number; downregulate the protein expression of caspase‐3, caspase‐9, TLR4, MyD88, and NF‐κB(p65) (p < .05); and inhibit the IL‐6, IL‐1β, and TNF‐α concentrations in serum and BALF. Houttuynia could improve the morphology and apoptosis cell number in lung tissues, thereby inhibiting the activation of the TLR4/MyD88/NF‐κB(p65) signaling pathway.

TNF-alpha Concentrations in Serum and Human Milk of Mothers With Insufficient Versus Sufficient Milk Production

ObjectivesObesity and poor metabolic health are associated with poor lactation outcomes, potentially driven by elevated tumor necrosis factor alpha (TNF-α) from chronic inflammation. TNF-α reduces lipoprotein lipase (LpL) activity, limiting triglyceride (TG) uptake by the mammary gland for milk synthesis. Our objective here is to determine if TNF-α concentrations in serum and milk are higher in mothers with severely low compared to sufficient milk production and if TNF-α is associated with plasma TG. MethodsMothers screened for low milk production were categorized into very low (VL; 177 [87,219] mL/d; n = 23) and moderately low/normal (ML; Median [IQR]: 467 [373,655] mL/d; n = 20) milk production. An external control group had sufficient milk production (EC; 783 [717,818] mL/d; n = 18). Mothers provided a spot milk sample at a morning clinic visit, where fasting blood samples were taken for clinical metabolic health measures. TNF-α concentrations were measured in duplicate by high-sensitivity ELISA in serum and aqueous milk. Group differences were assessed by Wilcoxon Rank Sums Tests or Fisher’s Exact Chi Square Test (α = 0.05). Correlations were assessed by Spearman rank order. ResultsMothers were 31.3 [27.3, 34.1] years old, delivered at gestational age of 39.4 [38.6, 40.3] weeks, and were 43 [35, 56] days postpartum, with no significant differences among groups. Milk TNF-α was detected in 47% of samples (1.3 [0.8, 2.2] pg/mL). The proportion of detectable TNF-α (D-TNF) in milk was higher in VL compared with ML and EC (71%, 32%, 33% respectively; p = 0.02). Compared to undetected (U-TNF), subjects with D-TNF in milk had higher plasma TG (D-TNF: 94 [68,140] mg/dL; U-TNF: 60 [49, 84] mg/dL; p = 0.001). Serum TNF-α was not significantly different by group (VL: 6.3 [5.8, 7.0] pg/mL; ML: 6.1 [5.6, 6.5]; EC: 5.8 [5.3, 6.6]; p = 0.31), but was correlated with plasma TG (ρ = 0.40, p = 0.002). ConclusionsIn this exploratory cohort, TNF- α levels were higher in milk from mothers with low milk production, and both milk and serum TNF- α was associated with higher plasma TG. This suggests that proximal TNF-α expression is more associated with reduced mammary LpL activity than circulating serum TNF-α, leading to low milk production. Funding SourcesNIH and USDA.

BIG1 mediates sepsis-induced lung injury by modulating lipid raft-dependent macrophage inflammatory responses.

Sepsis is a systemic inflammatory response syndrome with high mortality. It has been reported that brefeldin A-inhibited guanine nucleotide-exchange factor 1 (BIG1) is involved in the pathogenesis of sepsis. However, the mechanism is not fully elucidated. In the present study, we explored the role of BIG1 in mediating lipid raft-dependent macrophage inflammatory response and its impact on lung injury in murine sepsis. In vitro studies revealed that BIG1 deficiency reduces the upregulation and secretion of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and IL-1β and inhibits the activation of the toll-like receptor 4 (TLR4)/myeloid differentiation primary response 88-dependent nuclear factor kappa-B signaling pathway induced by the lipopolysaccharide (LPS) treatment. Further experiments revealed that the inhibitory effects of BIG1 deficiency on LPS-induced inflammation are due to the upregulation of adenosine triphosphate-binding cassette transporter A1. This promotes the free-cholesterol efflux from lipid rafts and results in the reduction of lipid raft TLR4 content. The decrease in TLR4 content in lipid raft thereby inhibits the LPS-induced inflammatory response. Furthermore, using the cecal ligation and puncture-induced polymicrobial sepsis mouse model, we found that conditional knockout (cKO) of the myeloid cell BIG1 significantly reduced the serum concentrations of TNF-α, IL-6, and IL-1β, and downregulated their mRNA expressions in the lungs. Pathological analysis confirmed that the BIG1 cKO alleviated the sepsis-induced lung injury. These results revealed the crucial new role of BIG1 in mediating lipid raft-dependent macrophage inflammatory response. Hence, BIG1 may be a potential promising therapeutic target for the treatment of septic lung injury.

MO031KLOTHO GENE PROMOTER METHYLATION IN THE VASCULATURE IS RELATED TO INFLAMMATION IN HUMAN ATHEROSCLEROSIS

Abstract Background and Aims Atherosclerosis, a vascular pathological process with an important inflammatory component, underlies most of the cardiovascular diseases (CVD). Deficiencies in the antiaging factor α-Klotho (KL) have been related to the appearance and progression of atherosclerotic damage associated with CVD. In addition, some studies have demonstrated that KL gene is expressed in human arteries and that low vascular KL expression levels are related with clinical atherosclerotic disease. Among the epigenetic mechanisms that regulate the expression of the KL gene, methylation of the CpG islands in the promoter region is one of the most studied. In this pilot study, we propose that this mechanism participates in the regulation of vascular KL expression and contributes to the deficiency in vascular KL levels observed in CVD. Moreover, we determined the influence of different inflammatory components in the methylation status of KL promoter. Method We developed a case-control study that included 25 patients with diagnosis of clinical atherosclerotic vascular disease undergoing elective vascular surgery and 15 cadaveric organ donors with no medical history of CVD, respectively. Vascular fragments were retrieved from all subjects, and the degree of methylation of the KL gene promoter was assessed by bisulfite sequencing. Additionally, vascular gene expression in both groups was assessed by qPCR for KL, two DNA methyltransferases (DNMT1 and DNMT3A), and three inflammatory-related loci (TNF, IL10 and NFKB1). Serum concentrations of TNFα and IL10 were measured by ELISA immunoassay, and complementary inflammatory parameters (CRP and neutrophil/lymphocyte ratio) by standardized routine methods. Results The vascular tissue of patients in the case group presented a higher percent level of methylated positions in the KL promoter region (71.4±8.3% vs. 39.4±12.3%, P&amp;lt;0.05), as well as a significant decrease in the expression of KL gene (a 40% reduction as compared to controls, P&amp;lt;0.05). Considering the whole study population, the methylation degree was inversely related to the endogenous transcript levels of KL gene (r=-0.66, P&amp;lt;0.0001). Moreover, the expression of DNMT1 was significantly increased in the case group (P&amp;lt;0.05), as well as those of the proinflammatory genes TNF and NFKB1 (P&amp;lt;0.0001 and P&amp;lt;0.05, respectively), being all of them directly associated with the vascular methylation levels of KL gene (r=0.51, P&amp;lt;0.0001 for DNMT1; r=0.45, P&amp;lt;0.001 for TNF; r= 0.37, P&amp;lt;0.05 for NFKB1). Furthermore, vascular expression of DNMT1 and DNMT3A directly correlated with both inflammatory markers in this tissue (P&amp;lt;0.0001, for all). Systemic levels of IL10 presented a direct association with vascular KL gene expression (r=0.28, P&amp;lt;0.05), while inverse associations were observed with the methylation degree of KL gene (r=-0.38, P&amp;lt;0.05) and DNMT1 expression (r=-0,29, P&amp;lt;0.05). Conclusion These results suggest that the methylation of KL gene promoter in the vasculature is a mechanism associated with the inflammatory response that would partly explain its reduction during atherosclerotic injury.

MO741TUMOR NECROSIS FACTOR ALPHA AND MAGNESIUM ARE LINKED TO CARDIAC VALVE CALCIFICATION IN DIABETIC HEMODIALYSIS PATIENTS

Abstract Background and Aims The processes of cardiac valve calcification (CVC) in diabetic hemodialysis (HD) patients are not fully understood. In this context, it is reasonable to complex and comprehensively research the activity of chronic inflammation and magnesium (Mg) imbalance as cardiovascular risk factors in end-stage renal disease (ESRD). The main purpose of the current study was to determine the relationship of tumor necrosis factor alpha (TNF-α) and Mg levels with the presence and severity of CVC in diabetic patients with ESRD. Method We enrolled 136 patients undergoing HD (male/female, 78/58;age, 53.9±1.0 years; HD duration, 47.6±4.2 month) in this observational cross-sectional study. The study was performed in accordance with the provisions of the Declaration of Helsinki last revision. Depending on the presence of type 2 diabetes mellitus (T2DM) all subjects were divided into two groups: the 1st one – non-diabetic patients (n=88); the 2nd one – diabetic patients (n=48). Presence of CVC was detected by ultrasound. The mitral (MVC) and aortic (AVC) valve calcification degree were scored as follows: 1, no calcification; 2, valve thickening without calcification; 3, valve annulus or cusps calcification. Serum content of TNF-α as one of the key proinflammatory cytokine was determined by enzyme-linked immunosorbent assay. Serum Mg concentration was estimated by biochemical method. Data are expressed as means±SEM. Used nonparametric statistics methods: Mann-Whitney U-test, χ2-test, Spearman’s rank R correlations. Results In diabetic HD patients TNF-α content was higher (13.86±1.34 vs. 8.73±0.60 ng/L; Z=3.04, p=0.002) whereas Mg concentration (0.87±0.02 vs. 1.00±0.02 mmol/L; Z=4.91, p&amp;lt;0.001) – lower compared to non-diabetic ones, and in 2nd group indices of TNF-α and Mg were related (Rs=-0.68, p&amp;lt;0.001). CVC was detected in 66.6% of T2DM patients with predominance of calcification of both valves (35.4%) over isolated MVC (20.8%) and AVC (10.4%). Combined MVC and AVC in the 2nd group was observed 2.6 times more often (χ2=8.78, p=0.003) than in the 1st one. For the first time it was established that in diabetic patients with ESRD the presence of CVC closely associated with indices of TNF-α (Rs=0.51, p&amp;lt;0.001) and Mg (Rs=-0.57, p&amp;lt;0.001). The MVC as well as AVC degree were related with the content of TNF-α (Rs=0.49, p&amp;lt;0.001; Rs=0.52, p&amp;lt;0.001) and Mg concentration (Rs=-0.47, p&amp;lt;0.001; Rs=-0.50, p&amp;lt;0.001) respectively. Conclusion (1) T2DM in HD patients is characterized with an increase of serum TNF-α activity and simultaneous decreased of Mg content. (2) In diabetic patients with ESRD, both MVC and AVC are closely linked with the TNF-α accumulation and hypomagnesemia. (3) Chronic inflammation and Mg deficiency can be important factors of CVC progression and very high cardiovascular risk in diabetic HD patients.

IL-1β, IL-10 and TNF-α polymorphisms may affect systemic lupus erythematosus risk and phenotype.

Systemic lupus erythematosus (SLE) is an autoimmune disease, and IL-1β, IL-10, and TNF-α genes are important in the pathogenesis of this disease. We studied the impact of IL-1β-511, IL-1β +3953, IL-10 -592, IL-10 -1082, TNF-α -308, TNF-α -238, and TNF-α +489 polymorphisms on SLE risk and phenotype in SLE patients and healthy controls. We genotyped SLE patients and healthy controls by real-time PCR on QuantStudio 5 (Applied Biosystems) and measured levels of cytokines by enzyme-linked immunosorbent assay (ELISA). We indicated that TNF-α -308, IL-10 -592, IL-10 -1082, IL-1β-511 and IL-1β +3953 polymorphisms affect SLE risk. Furthermore, we exposed that some of the TNF-α +489, TNF-α -238, IL-10 -1082 and IL-1β +3953 genotypes are connected with the SLE phenotype. Moreover, we discovered the linking between specific genotypes and the serum concentrations of TNF-α, IL-1β, and IL-10. In conclusion, our study revealed that IL-1β-511, IL-1β +3953, IL-10 -592, IL-10 -1082, and TNF-α -308 polymorphisms may affect SLE risk and phenotype.

IFN-α and TNF-α serum levels and their association with disease severity in Egyptian children and adults with alopecia areata.

Alopecia areata (AA) is an autoimmune skin disease characterized by abnormal levels of several cytokines, such as interferon alpha (IFN-α) and tumor necrosis factor-alpha (TNF-α), which are T-helper type 1 cytokines that have important roles in the pathogenesis of AA. The aim of our study was to correlate circulating IFN-α and TNF-α levels with disease severity, activity, and clinical type in patients with AA and to evaluate the relationship between the two cytokines. We investigated serum IFN-α and TNF-α levels in 72 patients with AA (35 children and 35 adults) and 75 healthy control individuals (34 children and 41 adults) using the enzyme-linked immunosorbent assay (ELISA) technique. We evaluated AA severity using the Severity of Alopecia Tool (SALT) and determined the activity based on dermoscopic criteria of disease activity. Serum IFN-α and TNF-α concentrations were significantly higher in the patients than in the controls. There was a significant positive correlation between serum IFN-α and TNF-α levels in all patients with alopecia areata, as well as between serum TNF-α levels and disease severity in all patients and in children. Our results support the association between IFN-α and TNF-α levels and AA and suggest that TNF-α might be related to disease severity.

Hemodynamic changes in cytokines, chemokines, acute phase proteins and prostaglandins in mares with subclinical endometritis

Apparently healthy mares with conception failure or embryonic loss may have subclinical endometritis (SE). Our objective was to document evidence of systemic immune responses against SE in mares. In apparently healthy (control) mares as well as those with chronic endometritis (ChE) or subacute suppurative endometritis (SSE), both considered classes of SE, serum concentrations of cytokines (IL-1β, IL-6, IL-10 and TNF-α), chemokines (IL-8), acute phase protein (SAA), and plasma concentrations of prostaglandins (PGF2α and PGE2) were measured using validated enzyme linked immunoassays (EIA). Mixed-breed mares of known reproductive history, were used. Based on an endometrial cytological examination, mares were allocated into the following groups: healthy (control), ChE, and SSE (12, 26 and 11 mares, respectively). Serum concentrations of IL-6, IL-8 and IL-10 and plasma PGF2α concentrations were increased (P < 0.01) in mares with SSE compared to healthy mares. Furthermore, serum concentrations of IL-6 (P < 0.05) and IL-10 (P < 0.05) were elevated in the ChE group compared to the healthy group whereas serum SAA (P < 0.01) and plasma PGE2 (P < 0.05) were higher in healthy mares compared to ChE and SSE mares. However, serum concentrations of IL-1β and TNF-α were not significantly different among groups. In conclusion, there were two novel findings: (1) development of serum testing for a set of biochemical markers has promise to explore the pathogenesis of inflammation in mares with SE; and (2) none of the individual biomarkers studied, excluding IL-8, was a significant predictor of SE. However, serum IL-6:IL-10 and IL-6:TNF-α concentrations, as well as plasma PGE2:PGF2α ratios, may yield a novel diagnostic marker for chronic subclinical endometritis.