TNF Concentrations Research Articles

Effects of resveratrol on reducing spermatogenic dysfunction caused by high-intensity exercise

BackgroundLong-term high-intensity exercise can lead to reproductive endocrine and spermatogenic dysfunction. This research is to investigate the effect of resveratrol on the reduction of reproductive dysfunction induced by high-intensity exercise, and to screen relevant factors and signal transduction pathways.MethodsRats were randomly divided into three groups, a control group, an intensive exercise group (IE group), and a resveratrol-treated group (RSV group). After 9 weeks of exercise, the sperm density and reproductive hormone concentrations were measured, along with antioxidation, inflammatory cytokine production, and histological analyses performed for each group. In addition, a proteomics analysis of the IE group and RSV group were conducted.ResultsWe found that compared with the control group, the average sperm density (P < 0.05) and testosterone concentration (P < 0.05) in the IE group decreased significantly. Additionally, in testis tissue the concentration of the inflammatory cytokines IL-6 (P < 0.01) and TNF-α (P < 0.01) increased significantly. Also, a significant decrease in superoxide dismutase (SOD) activity (P < 0.01) and a significant increase in the malondialdehyde (MDA) concentration (P < 0.01) were noted. In the RSV group, the average sperm density (P < 0.01), testosterone (P < 0.01) and follicle stimulating hormone (FSH) levels (P < 0.01) all increased in comparison to the IE group, and the concentration of IL-6 (P < 0.01) and TNF-α (P < 0.01) were found to be significantly decreased. Compared with the IE group, the SOD activity in the RSV group was significantly increased (P < 0.01), while the MDA content decreased (P < 0.01). Furthermore, histological analysis showed that the number of spermatogenic epithelial cells in the RSV group was higher than that of the IE group. There were a number of spermatogenic regulatory proteins identified in the proteomics analysis, including Clusterin, Piwi like homolog 1 (Piwil1), Zona pellucida binding protein (Zpbp), Heat shock-related 70 kDa protein 2 (Hspa2), Centrin 1, and Bardet-Biedl syndrome 2 protein (Bbs2). It was found that the proteins that differed between the two groups were mainly involved in pathways such as complement and coagulation cascades, the extracellular matrix-receptor interactions, etc.ConclusionsThe present study demonstrates that after high-intensity exercise, the inflammatory cascade in the tissue of the testis increases with decreased resistance to oxidation and disordered spermatogenic function. Resveratrol can improve the reproductive dysfunction of rats that was induced by high-intensity exercise. It mostly promotes reproductive function by increasing testosterone secretion, reducing the inflammatory response, improving the antioxidant capacity, affecting the expression of spermatogenic regulatory proteins, and enhancing the signal transduction pathway of spermatogenesis.

Pituitary Adenylate Cyclase-Activating Polypeptide-A Neuropeptide as Novel Treatment Option for Subacute Ileitis in Mice Harboring a Human Gut Microbiota.

The neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP) is well-known for its important functions in immunity and inflammation. Data regarding anti-inflammatory properties of PACAP in the intestinal tract are limited, however. In our present preclinical intervention study we addressed whether PACAP treatment could alleviate experimental subacute ileitis mimicking human gut microbiota conditions. Therefore, secondary abioitic mice were subjected to human fecal microbiota transplantation (FMT) and perorally infected with low-dose Toxoplasma gondii to induce subacute ileitis on day 0. From day 3 until day 8 post-infection, mice were either treated with synthetic PACAP38 or placebo. At day 9 post-infection, placebo, but not PACAP treated mice exhibited overt macroscopic sequelae of intestinal immunopathology. PACAP treatment further resulted in less distinct apoptotic responses in ileal and colonic epithelia that were accompanied by lower T cell numbers in the mucosa and lamina propria and less secretion of pro-inflammatory cytokines in intestinal ex vivo biopsies. Notably, ileitis-associated gut microbiota shifts were less distinct in PACAP as compared to placebo treated mice. Inflammation-ameliorating effects of PACAP were not restricted to the intestines, but could also be observed in extra-intestinal including systemic compartments as indicated by lower apoptotic cell counts and less pro-inflammatory cytokine secretion in liver and lungs taken from PACAP treated as compared to placebo control mice, which also held true for markedly lower serum TNF and IL-6 concentrations in the former as compared to the latter. Our preclinical intervention study provides strong evidence that synthetic PACAP alleviates subacute ileitis and extra-intestinal including systemic sequelae of T cell-driven immunopathology. These findings further support PACAP as a novel treatment option for intestinal inflammation including inflammatory bowel diseases (IBD).

Complex analysis of correlation interrelations between indicators of humoral and cellular immunity in persons vaccinated against plague

At present, the search for biomarkers, which indicate the presence of tense antiplague immunity, is continuing. The aim of the study was to determine the parameters of cellular and humoral immunity in persons vaccinated with the plague vaccine according to epidemiological indications and to characterize the correlation interrelations between them in the dynamics of the antiplague response formation. Blood samples from 114 people living in the territory of the Pre-Caspian sandy natural plague foci were investigated. Using commercial kits, the production of cytokines IL-1, IL-2, IL-4, IL-6, IL-8, IL-10, IL-18, IFN, TNF (Vector-Best, Russia) IL-17A (eBioscience, Austria), immunoglobulins IgG, IgM, IgA (Vector-Best, Russia) was determined on the enzyme immunoassay analyzer LAZURIT (Dynex Technologies, США). Blood leukocyte immunophenotyping was carried out using f luorochrome-labeled monoclonal antibodies Cyto-Stat CD45-FITC, CD4-PE, CD8-ECD, CD3-PC5 (Beckman Coulter, США) on the CyAn ADP DakoCytomation f low cytometer (Denmark). Obtained data analysis revealed a number of differences in the production of detectable cytokines (TNF, IL-1, IL-2, IL-4, IL-6, IL-17) between groups of vaccinated and first-vaccinated people. However, human blood cells, regardless of the multiplicity of the graft, retained their functional reactivity and were able to actively secrete the majority of the analyzed cytokines (8 of 10) in response to stimulation with concanavalin A. Diagnostically significant changes in the concentration of IFN, TNF and IL-4 were detected for the characteristics of a specific antiplague response. For these marker cytokines, the greatest number of correlations have been revealed, both with each other and with changes in the level of other parameters: cytokines, immunoglobulins, T cells. The revealed multiple correlation links between the cytokine and immune status indices indicate the activation of both the cellular and humoral components of the antiplague immune response. The informative value of the complex analysis application of correlation interrelations between the humoral and cellular immunity parameters is shown to assess the level of immunological efficacy (actual vaccination) of vaccination in persons vaccinated against plague.

Tumor necrosis factor-alpha and Fas/Fas ligand signaling pathways in chronic spontaneous urticaria

BackgroundThere is increasing evidence pointing to the important role of tumor necrosis factor-alpha (TNF-α), a key inflammatory and apoptotic mediator in urticarial inflammation. However, the role of the TNF-α system and Fas/Fas ligand (FasL) in the apoptosis-inducing pathways in chronic spontaneous urticaria (CSU), remain unclear.AimTo determine circulating concentrations of TNF-α, soluble TNF-α receptor type 1 and type 2 (sTNF-R1 and sTNF-R2, respectively) as well as soluble Fas (sFas) and FasL (sFasL) in CSU subjects.MethodsSerum TNF-α, sTNF-R1, sTNF-R2, sFas, sFasL concentrations were measured using enzyme-linked immunosorbent assay in CSU subjects and in the healthy subjects.ResultsTNF-α concentrations were significantly higher in CSU subjects and moderate-to-severe CSU than in the controls, while there were no significant differences in TNF-α concentrations between subjects with mild CSU and the controls. sTNF-R1 and sTNF-R2 concentrations were significantly higher in all CSU and moderate-severe CSU subjects vs. the controls. Serum concentrations were also significantly higher in mild CSU vs. the controls, but not in moderate-severe CSU vs. mild CSU. No significant differences were observed in sFas and sFasL concentrations between CSU subjects and the healthy controls. Significant correlations were found between concentrations of TNF-α and its receptors, as well as sTNF-R1 and sTNF-R2, but not with the urticaria activity score (UAS). There was no relationship between TNF-α/sTNF-R1/sTNF-R2 and sFas/sFasL pathways in CSU.ConclusionsCSU is associated with the activation of the TNF-α/receptors signaling pathway, marked by increased circulating concentrations of TNF-α, sTNF-R1 and sTNF-R2, which are related to each other in this disease. In contrast, the circulating sFas/FasL system is not up-regulated in CSU, and sFas/sFasL may not be a useful marker of the activity/severity of urticarial processes. Considering the lack of significant changes in sFas/sFasL (mainly reflecting systemic apoptosis) in CSU patients, it appears that elevated serum TNF-α concentrations are related to its pro-inflammatory function rather than an enhanced systemic apoptotic response in CSU.

Bronchoalveolar lavage cell profiles and proteins concentrations can be used to phenotype extrinsic allergic alveolitis patients

BackgroundExtrinsic allergic alveolitis (EAA) patients form heterogenous group with different clinical manifestation and different prognosis. We aimed to determine how to phenotype distinct EAA subgroups. Predictive role of the bronchoalveolar lavage fluid (BALF) IL-4Rα concentration at the time of diagnosis with regard to the clinical behavior in EAA patients was studied.MethodsConcentrations of MMP-7, IL-4Rα, TNF-α, and PAR-2 were measured in the BALF od 71 EAA patients at the time of diagnosis. Lung functions and outcome data were assessed at 12 months after the diagnosis. Correlations between the BALF protein concentration, cell profile, lung functions and patient outcome were determined.ResultsWe found positive correlations between BALF IL-4Rα concentration and BALF eosinophils (p = 0,006), negative correlation between IL-4Rα BALF concentration and diffusing capacity (DLco) (p = 0,003), negative correlation between IL-4Rα BALF concentration and forced vital capacity (FVC) (p = 0,004) and negative correlation between IL-4Rα concentration and BALF lymphocytes (p = 0,04). The BALF concentration of IL-4Rα was significantly higher in acute exacerbation patients (p = 0,0032) and in patients progressing despite corticosteroid treatment (p = 0,04). We observed a positive correlation between MMP-7 BALF concentration and the BALF lymphocytes (p = 0.05), negative correlation between the PAR-2 BALF concentration and DLco (p = 0.04) and a negative correlation between the BALF TNF-α concentration and DLco (p = 0.03).ConclusionsSpecific subgroup of EAA patients with more severe functional impact, distinct BALF cell profile and higher IL-4Rα BALF concentration can be differentiated. Correlations between the BALF concentrations of PAR-2, MMP-7 and TNF-α with clinical parameters may reflect the role of inflammation in the pathogenesis of EAA.

Does aerobic exercise associated with tryptophan supplementation attenuates hyperalgesia and inflammation in female rats with experimental fibromyalgia?

The objective of this study was to verify the effects of aerobic exercise associated with tryptophan (TRP) supplementation on hyperalgesia, as well as on cortisol, IL-6 and TNF concentrations in female rats with experimental fibromyalgia (FM). Female Wistar rats (initial body weight: ~ 350 g; age: 12 months) were randomly divided into 5 groups: CON (Control); F (Fibromyalgia induced); FE (Fibromyalgia induced plus exercise); FES (Fibromyalgia induced plus exercise and TRP supplementation) and FS (Fibromyalgia induced plus TRP supplementation). Fibromyalgia was induced with two injections (20 μL) of acidic saline (pH 4.0) into the right gastrocnemius muscle with a 3-day interval. Control animals received the same doses of neutral saline (pH 7.4). The exercised animals underwent progressive low-intensity aerobic exercise (LIAE) on a treadmill (10–12 m/min, 30–45 min/day, 5 days/week) for three weeks. During this period, the supplemented animals received a TRP supplemented diet (210 g/week), while the others received a control diet. Mechanical hyperalgesia was evaluated weekly and serum cortisol and muscle IL-6 and TNF concentrations were assessed after three weeks of interventions. Experimental FM caused bilateral hind paw hyperalgesia and augmented serum cortisol and muscle IL-6 concentrations. After 3 weeks of interventions, LIAE alone reduced hyperalgesia (151%) and reduced serum cortisol concentrations (72%). Tryptophan supplementation itself diminished hyperalgesia (57%) and reduced serum cortisol concentrations (67%). Adding TRP supplementation to LIAE did not further reduce hyperalgesia significantly (11%), which was followed by an important decrease in muscle IL-6 concentrations (68%), though reduction in serum cortisol pulled back to 45%. Muscle TNF concentrations were not affected. In conclusion, the association of TRP supplementation to LIAE does not potentiate significantly the reduction of bilateral mechanical hyperalgesia promoted by LIAE in female rats with experimental FM, however an important decrease in IL-6 is evident.

ANTIHYPERGLYCEMIC, ANTIOXIDANT AND ANTI-INFLAMMATORY EFFECTS OF AQUEOUS EXTRACT OF MISTLETOE (Cladocolea loniceroides) IN STZ-INDUCED DIABETIC MICE

Background: Inhibition of carbohydrate hydrolyzing enzymes, such as α-amylase and α-glucosidase, is a key element in the regulation of diabetes mellitus (DM). The purpose of this work was to study the inhibition of carbohydrate hydrolyzing enzymes, and the antihyperglycemic activity of aqueous extract of Cladocolea (C. loniceroides) in streptozotocin (STZ)- induced diabetic mice. Materials and Methods: The inhibitory activities of C. loniceroides aqueous extract on α-amylase and α-glucosidase were investigated in vitro. Glucose tolerance test was performed in normoglycemic (NG) mice which were fed with starch or sucrose. The effect of mistletoe aqueous extract (ME) was measured in (STZ)-induced diabetic mice. On day 35 of the treatment, the effect of decreasing oxidative stress (lipid peroxidation, glutathione redox state, GPx and GR specific activities, cytokines and aminotransferases analysis) was assessed. Results: ME showed a competitive mode of inhibition for the carbohydrate hydrolyzing enzymes (CHE). The maximum antihyperglycemic activity in mice was observed for the unripe fruit aqueous extract (UFAE) for -amylase and stem aqueous extract (SAE); for -glucosidase due to the glycemic response reduction by 23% or 35%, respectively. UFAE decreased malondialdehyde (MDA) 1.76 times; GSH/GSSG ratio was mantained (3.08 ± 0.66); GPx activity was reduced (24%); IL-6 was reduced (18%) and the concentration of TNF- (37%) was leveled with respect to the (STZ)-induced diabetic mice; ALT and AST (liver transaminases) levels were nearly the same compared with those found in the NG mice. Conclusion: UFAE of C. loniceroides exhibited the highest antidiabetic activity in (STZ)-induced diabetic mice.

Fecal sST2 correlates with the disease severity of ulcerative colitis

Background/Aim. Ulcerative colitis (UC) is a chronic, relapsing inflammatory disease affecting the distal colon and rectum with complex pathogenesis and diagnosis, indicating the need for new diagnostic and prognostic markers. The aim of this study was to determine the fecal values of TNF- ?, IL-17, IL-10 and soluble protein ST2 (sST2) in the patients with UC and their relationship with clinicopathological aspects. Methods. The samples of stool of 80 patients with UC were analyzed. Concentrations of TNF-?, IL-17, IL-10 and sST2 were measured by ELISA. Results. Concentrations of TNF-?, IL-17 and sST2 were significantly increased in the feces of patients with the higher endoscopic, clinical and total Mayo score, as well as in the patients with an intense crypt destruction, erosion of the mucous membranes, architectural changes, neutrophil infiltration and eosinophil infiltration. The local value of anti-inflammatory cytokine IL-10 in liquid fraction of feces was increased in the patients with an advanced endoscopic stage of UC. The moderate positive correlation between the fecal sST2/IL-17 and the clinical and histological parameters of disease severity and also the strong correlation between sST2 and IL-17 was also observed in the feces of patients with UC. The analysis of receiver operating characteristic (ROC) curves showed that the optimal cut-off value for sST2 of 624.0 pg/g allows the discrimination of clinical stages of UC. Conclusion. The increased fecal value of sST2 in the UC patients with a higher endoscopic, clinical and histological stage of disease may be considered as a sign of the disease severity. The fecal values of sST2 can be used as a valuable marker for UC severity.

Preoperative Immune Response is Associated with Perioperative Transfusion Requirements in Glioma Surgery.

Immunosuppression induced by transfusion causes postoperative adverse events including poor prognosis in cancer, but data on influence of the immune response on blood transfusion requirements during perioperative period are limited. The aim of this study was to investigate whether the preoperative immune response is associated with perioperative blood cell transfusion in a glioma surgical patient population. The authors identified 321 cases of surgery for treatment of glioma. Patient variables, preoperative laboratory variables (hemoglobin, platelet count, activated partial thromboplastin time, prothrombin time, hematocrit, red and white blood cell count), and transfusions were registered. Plasma concentration of Th-associated cytokines was measured by flow cytometry. Multivariable regression analysis and receiver operating characteristic curve were undertaken to identify predictors of transfusion. Of 321 patients, 157 (48.90%) received red blood cells transfusion. The mean age is significantly higher in transfusion group compared to no transfusion group, while postoperative hospital stay, preoperative hemoglobin, prothrombin time, activated partial thromboplastin time, platelet count, red and white blood cell count and hematocrit of patients did not differ significantly between the two groups. No significant differences of IL-2, -4, -6, -10 and INF-γ concentration were observed between transfusion and no transfusion group. The concentration of TNF and IL-17A was significantly lower in transfusion patients than in the no transfusion subjects. Low plasma TNF and IL-17A levels predicted high perioperative transfusion rate, the combination of them enlarged the prognostic accuracy of testing. Our study demonstrates that the preoperative immune response influences transfusion requirements, and TNF and IL-17 are important predictive risk factors for perioperative use of blood components in glioma patients.

Hypogonadism in chronic obstructive pulmonary disease (COPD): Risk factors

Bacground/Aim. Chronic obstructive pulmonary disease (COPD) is the leading cause of morbidity and mortality in pulmonary pathology. However, apart from its own pulmonary manifestations, this disease is also characterized by systemic effects, including hypogonadism which is described especially in the group of men with COPD. The aim of this study was to evaluate risk factors for hypogonadism in men with COPD. Methods. The study included 96 male patients with COPD in stable phase of the disease. All patients were checked for concentration of free testosterone in serum, markers of systemic inflammation, tumor necrosis factor-? (TNF-?), interleukin-1? (IL-1?) and C reactive protein (CRP), pulmonary function test, gas exchange parameters, a 6-minute walk test (6MWT), nutritional status and condition of skeletal muscle (midthigh muscle cross-sectional area ? MTCSA using computed tomography). Results. Decreased value of free testosterone was found in 37.5% of the patients. In the group with hypogonadism (free testosterone &lt; 4.5 pg/mL), we found significantly increased serum concentration of TNF-? (5.88 ? 3.21 vs. 3.16 ? 2.53 pg/mL; p &lt; 0.05), significantly lower MTSCA (68.2 ? 18.72 vs. 91.1 ? 21.4 cm2; p &lt; 0.05) and the 6MWT (268.33 ? 32.35 vs. 334.25 ? 43.25 m; p &lt; 0.05). Lung function, gas exchange markers and body mass index (BMI) were similar in both groups. The multivariate regression analysis singled out serum value of TNF-? as an independent predictor of serum concentrations of free testosterone (B = -0.157; 95% confidence interval: -0.262?0.053). Conclusion. In our analysis we found that TNF-? as a marker of systemic inflammation is an independent predictor of the presence of hypogonadism in the patients with COPD. Our results indicate that hypogonadism predisposes to skeletal muscle wasting and exercise intolerance in male COPD patients.

Brain swelling is independent of peripheral plasma cytokine levels in Malawian children with cerebral malaria

BackgroundCerebral malaria (CM) is often fatal, and severe brain swelling is a predictor of CM-related mortality. CM is characterized by elevated circulating pro-inflammatory cytokines TNF and IFN-γ and anti-inflammatory cytokine IL-10, however whether cytokine levels correlate with brain swelling severity is unknown. This study therefore was conducted to investigate the relationship between cytokine levels and brain swelling severity in children presenting with CM.MethodsA total of 195 Malawian children presenting with CM were recruited and had the concentrations of plasma cytokines determined and compared to brain swelling severity, determined by MRI examination, and graded as severe, moderate, mild or none.ResultsLevels of IL-1β, IL-6, IL-8 and IL-10 did not differ between CM patients with and without severe brain swelling. Compared to children without brain swelling, IL-12 levels were higher in children with severe swelling (p < 0.01, no swelling 1 pg/mL, IQR [1] vs. severe swelling 18.7 pg/mL, IQR [1–27]), whereas TNF concentrations were higher in children with moderate brain swelling compared to children with no swelling (p < 0.01, no swelling 3 pg/mL, IQR [1–20] vs. moderate swelling 24 pg/mL, IQR [8–58]. Multivariate analysis showed that no single cytokine independently predicted brain swelling.ConclusionSevere brain swelling in paediatric CM was independent of tested blood pro-inflammatory and anti-inflammatory cytokines which are markers of systemic inflammation.

Immunization of Experimental Dogs With Salivary Proteins From Lutzomyia longipalpis, Using DNA and Recombinant Canarypox Virus Induces Immune Responses Consistent With Protection Against Leishmania infantum.

Metacyclic Leishmania promastigotes are transmitted by sand flies that inject parasites and saliva into the host's skin. Previous studies have demonstrated that DNA plasmids encoding Lutzomyia longipalpis salivary proteins LJM17 and LJL143, when used to immunize dogs, resulted in a systemic and local Th1 cell-mediated immunity that interfered in parasite survival in vitro. Here we evaluated the ability of these same salivary antigens to induce anti-Leishmania immunity and to confer protection by immunizing dogs using a novel vaccination strategy more suitable for use in the field. The strategy consisted of a single dose of plasmid followed by two doses of recombinant Canarypoxvirus (rCanarypoxvirus) expressing L. longipalpis salivary proteins (LJM17 or LJL143). Thirty days after the final immunization, dogs were intradermally challenged with 107 Leishmania infantum promastigotes in the presence of L. longipalpis saliva. We followed the experimentally infected dogs for 10 months to characterize clinical, parasitological, and immunological parameters. Upon vaccination, all immunized dogs presented strong and specific humoral responses with increased serum concentrations of IFN-γ, TNF, IL-7, and IL-15. The serum of dogs immunized with LJM17 also exhibited high levels of IL-2, IL-6, and IL-18. L. infantum infection was established in all experimental groups as evidenced by the presence of anti-Leishmania IgG, and by parasite detection in the spleen and skin. Dogs immunized with LJM17-based vaccines presented higher circulating levels of IFN-γ, IL-2, IL-6, IL-7, IL-15, IL-18, TNF, CXCL10, and GM-CSF post-infection when compared with controls. Results demonstrated that relevant Leishmania-specific immune responses were induced following vaccination of dogs with L. longipalpis salivary antigen LJM17 administered in a single priming dose of plasmid DNA, followed by two booster doses of recombinant Canarypox vector. Importantly, a significant increase in pro-inflammatory cytokines and chemokines known to be relevant for protection against leishmaniasis was evidenced after challenging LJM17-vaccinated dogs as compared to controls. Although similar results were observed following immunization with LJL143, the pro-inflammatory response observed after immunization was attenuated following infection. Collectively, these data suggest that the LJM17-based vaccine induced an immune profile consistent with the expected protective immunity against canine leishmaniosis. These results clearly support the need for further evaluation of the LJM17 antigen, using a heterologous prime-boost vaccination strategy against canine visceral leishmaniosis (CVL).

Boiling Histotripsy Ablation of Renal Cell Carcinoma in the Eker Rat Promotes a Systemic Inflammatory Response

Boiling histotripsy (BH) is an experimental focused ultrasound technique that produces non-thermal mechanical ablation. We evaluated the feasibility, short-term histologic effects and the resulting acute inflammatory response to BH ablation of renal cell carcinoma (RCC) in the Eker rat. Genotyped Eker rats were monitored for de novo RCCs with serial ultrasound (US) imaging. When tumors were ≥8 mm, rats underwent ultrasound-guided extracorporeal ablation of the tumor with BH, a pulsed focused US technique that produces non-thermal mechanical ablation of targeted tissues, or a sham US procedure. Treatments targeted approximately 50% of the largest RCC with a margin of normal kidney. BH treated rats were euthanized at 1 (n = 4) or 48 (n = 4) h, and sham patients (n = 4) at 48 h. Circulating plasma cytokine levels were assessed with multiplex assays before and at 0.25, 1, 4, 24 and 48 h following treatment. Kidneys were collected and processed for histologic assessment, immunohistochemistry and intrarenal cytokine concentration measurements. For statistical analysis Student's t-test was used. US-guided BH treatment was successful in all animals, producing hypoechoic regions within the targeted volume consistent with BH treatment effect. Grossly, regions of homogenized tissue were apparent with evidence of focal intra-parenchymal hemorrhage. Histologically, BH produced a sharply demarcated region of homogenized tumor and non-tumor tissue containing acellular debris. BH treatment was associated with significantly increased relative concentration of plasma TNF versus sham treatment (p < 0.05) and transient elevations in high-mobility group box 1 (HMGB1), IL-10 and IL-6 consistent with acute inflammatory response to trauma. Intrarenal cytokine concentrations followed the same trend. At 48 h, enhanced infiltration of CD8+ T cells was observed by immunohistochemistry in both the treated and un-treated contralateral RCC/kidneys in BH-treated animals versus sham treatment. BH treatment was well tolerated with transient gross hematuria and a perinephric hematoma developing in one subject each. The study demonstrates the feasibility of BH ablation of de novo RCC and suggests activation of the acute inflammatory cascade following treatment that appears to stimulate CD8+ T cell infiltration of both treated and untreated tumors. Longer duration chronic studies are ongoing to characterize the longevity and robustness of this response.

Influence of static forces on the expression of selected parameters of inflammation in periodontal ligament cells and alveolar bone cells in a co-culture in vitro model

ObjectiveAim of this study was to investigate the impact of human PDL-derived fibroblasts (HPDF) and human alveolar bone-derived osteoblasts (HABO) co-culture on the expression of cytokines involved in tissue remodeling using an in vitro compressive force (CF) model.Materials and methodsStatic compressive force (CF) of 47.4 g/cm2 was applied on mono- and co-cultured HPDFs and HABOs for 1, 2, or 4 h at 30 °C. TNFA, PTGS2, and IL6 gene expressions were determined by quantitative real-time polymerase chain reaction. TNF, PGE2, and IL6 concentrations were measured using enzyme-linked immunosorbent assay.ResultsIn mono-culture, TNFA, PTGS2, and IL6 gene expressions were upregulated under CF as compared to controls for each time period in both cell types. PGE2 increased at 1 and 2 h in both cell types, and IL6 increased only at 2 and 4 h in HPDFs. Co-culture alleviated the force-induced increase of the expression of TNFA, PTGS2, IL6, PGE2, and IL6 in HPDFs at any time point. In HABOs, co-cultivation decreased the expression of PGE2 after 1 h and 4 h, and that of IL6 after 1 h compared to mono-culture.ConclusionsCF application on co-cultures of HPDFs and HABOs causes significant changes of TNFA, PTGS2, and IL6 gene expressions and PGE2 and IL6 production in comparison to mono-culture indicating intercellular communication.Clinical relevanceMechanical stimulation of HPDFs and HABOs in co-culture induces a different gene expression pattern than stimulation of individual cell types alone. Co-culture might therefore be a relevant method to elucidate periodontal regeneration during orthodontic therapy.

The role of suppressor of cytokine signaling SOCS7 in the regulation of the phosphorylation of inhibitor of nuclear transcription factor NF-KB in mononuclear leukocytes and production of cytokines in community-acquired bacterial pneumonia

The study discusses the relationship between the content of NF-KB and cytokine signaling suppressor 7 (SOCS7) in mononuclear peripheral blood cells (MNC) phosphorylated form of nuclear transcription factor inhibitor (NF-KB) and the production of MNC cytokines (TNF, IFN, IL-1β, IL-4, IL-10, IL-12) determining the state of congenital and adaptive immune response.The content and level of phosphorylation of the nuclear transcription factor NF-KB (Ikba) inhibitor and the SOCS7 protein concentration were determined by enzyme immunoassay in MNC. In addition, the concentration of TNF, IFN, IL-1β, IL-4, IL-10, IL-12 was determined in cellular supernatants. The interrelations between the studied factors were evaluated by the method of linear regression analysis.The results of the study indicate that the stage of recovery of community-acquired pneumonia is accompanied by a decrease in the level of IL-1, TNF and IL-4 and an increase in the production of Information. Also in the stage of convalescence there is a decrease in phosphorylation of Ikba and an increase in the concentration of SOCS7 in the OLS. The analysis revealed a significant effect on the level of phosphorylation of Ikba content in the cell SOCS7. Thus, a strong negative relationship between SOCS7 and phosphorylation of Ikba can be mediated by inhibition under its influence of STAT3/5 and MARK / SAPK-dependent cytokine production mechanisms, which allows to consider this factor as a therapeutic target for limiting excessive immunosuppression in pneumonia.

Inhibition of RANKL-stimulated osteoclast differentiation by Schisandra chinensis through down-regulation of NFATc1 and c-fos expression

BackgroundSchisandra chinenesis (SC) has been reported to have ameliorative effect on osteoporosis. However, the mechanisms underlying the anti-osteoporosis activity of SC have not been clearly elucidated. In the present study, we determined the effects of SC on The receptor activator of NF-kB ligand (RANKL)-induced osteoclastogenesis and its potential mechanism.MethodsRaw 264.7 cells were treated with 0.6, 6 and 60 μg/mL SC in the presence of 100 ng/mL RANKL for 7 days. RANKL-induced osteoclast formation was analyzed by tartrate resistant acid phosphatase (TRAP) staining. The osteoclast differentiation-related factors were confirmed along with TNF-α.ResultsSC inhibits the RANKL-induced osteoclast differentiation in dose-dependent manner within non-toxic concentrations. The supernatant concentrations of TNF-α were significantly decreased by SC treatment. In addition, osteoclastogenesis-related factors, TRAP6 and NF-κB, were markedly decreased by SC in RANKL-induced osteoclasts. Mechanistically, SC reduced the RANKL-triggered NFATc1 and c-fos expressions.ConclusionsTaken together, our data suggest that SC can modulate bone metabolism by suppressing RANKL-induced osteoclast differentiation.

171. Association between adverse maternal clinical outcomes and imbalance of cytokines and angiogenic factors in preterm preeclampsia

Introduction Preeclampsia (PE) is a pregnancy specific syndrome, characterized by abnormal levels of cytokines and angiogenic factors, suggesting that immunological factors play a role in the disease development. Objective The objective of this study was to determine whether immunological markers are associated with the gestational age and with the disease severity at the time of the disease diagnosis. Methods Ninety-five women who developed PE were stratified for gestational age as preterm PE ( α ) and Interleukin 10 (IL-10) were determined by immunoenzymatic assay (ELISA). Results were analyzed by non-parametric test with significance set at p Results The comparison between preterm and term PE showed higher percentage of severe cases in the preterm PE (76.7%) than in the term PE (23.3%) group. Similarly the concentrations of TNF- α , TNF- α /IL-10 ratio, sFlt-1, sEng and sFlt-1/PlGF ratio were significantly higher in the preterm PE group. On the other hand, concentrations of PlGF, VEGF and IL-10 were significantly lower in the preterm PE group. Negative correlations between TNF- α and IL-10 (r = −0.5232; p Discussion Pregnant women with preterm PE showed an imbalance between immunological markers, with predominance of anti-angiogenic factors and TNF- α , associated with adverse maternal clinical outcomes.

Trovafloxacin-Induced Liver Injury: Lack in Regulation of Inflammation by Inhibition of Nucleotide Release and Neutrophil Movement.

The fluoroquinolone trovafloxacin (TVX) is associated with a high risk of drug-induced liver injury (DILI). Although part of the liver damage by TVX+TNF relies on neutrophils, we have recently demonstrated that liver recruitment of monocytes and neutrophils is delayed by TVX. Here we show that the delayed leukocyte recruitment is caused by a combination of effects which are linked to the capacity of TVX to block the hemichannel pannexin 1. TVX inhibited find-me signal release in apoptotic HepG2 hepatocytes, decelerated freshly isolated human neutrophils toward IL-8 and f-MLF, and decreased the liver expression of ICAM-1. In blood of TVX+TNF-treated mice, we observed an accumulation of activated neutrophils despite an increased MIP-2 release by the liver. Depletion of monocytes and neutrophils caused increased serum concentrations of TNF, IL-6, and MIP-2 in TVX-treated mice as well as in mice treated with the fluoroquinolone levofloxacin, known to have a lower DILI-inducing profile. This supports the idea that early leukocyte recruitment regulates inflammation. In conclusion, disrupted regulation by leukocytes appears to constitute a fundamental step in the onset of TVX-induced liver injury, acting in concert with the capability of TVX to induce hepatocyte cell death. Interference of leukocyte-mediated regulation of inflammation represents a novel mechanism to explain the onset of DILI.

Circulatory levels of tumor necrosis factor-α, interleukin-6, and syncytiotrophoblast microvesicles in the first trimester of pregnancy

Background: The first trimester of pregnancy remains an unknown immunological adaptation. Cytokines and syncytiotrophoblast microvesicles (STBM) have been identified as key factors involved in the maternal immune adaptation. This study therefore aims to determine the levels of T helper cell 1 (Th1) and 2 (Th2) associated cytokines TNF-&#x03B1; and IL-6 in relation to the relative concentration of STBM's in maternal circulation. Materials and Methods: Plasma samples from normotensive pregnant women in the first trimester of pregnancy were obtained. The concentrations of TNF-&#x03B1; and IL-6 were determined using ELISA. STBMs were determined by isolating microvesicles in maternal circulation and quantifying the concentration of PLAP using ELISA. Results: TNF-&#x03B1;, IL-6 and STBMs remained at constant levels in weeks 5-10 of gestation. In weeks 11-12 of gestation, TNF-&#x03B1; increased with a decrease in IL-6 and STBMs. Ratio of TNF-&#x03B1;/IL-6 remained constant in weeks 5-6 and significantly increased in weeks 11-12 of gestation. A positive correlation between TNF-&#x03B1; and IL-6 was obtained in weeks 6-10 and a negative correlation in weeks 11-12 of gestation. In addition, a positive correlation between STBMs & TNF-&#x03B1; was obtained and negative correlation between STBMs and IL-6. was observed Conclusion: The relationship between TNF-&#x03B1;/IL-6 and STBMs suggests that syncytiotrophoblast microvesicles may have a role in cytokine production and in the maintenance of the Th1/Th2 immune adaptation in normal pregnancy.

Host factors determine the evolution of infection with Staphylococcus aureus to gangrenous mastitis in goats

Staphylococcus aureus is the major cause of very severe mastitis of dairy goats. The initial objective of our study was to fine-tune an experimental model of infection of the goat mammary gland with two strains of S. aureus and two lines of goats (low and high somatic cell score lines). Following the challenge, the 10 infected goats divided in two clear-cut severity groups, independently of the S. aureus strain and the goat line. Five goats developed very severe mastitis (of which four were gangrenous) characterized by uncontrolled infection (UI group), whereas the other five kept the infection under control (CI group). The outcome of the infection was determined by 18 h post-infection (hpi), as heralded by the bacterial milk concentration at 18 hpi: more than 107/mL in the UI group, about 106/mL in the CI group. Leukocyte recruitment and composition did not differ between the groups, but the phagocytic killing at 18 hpi efficiency did. Contributing factors involved milk concentrations of α-toxin and LukMF′ leukotoxin, but not early expression of the genes encoding the pentraxin PTX3, the cytokines IL-1α and IL-1β, and the chemokines IL-8 and CCL5. Concentrations of TNF-α, IFN-γ, IL-17A, and IL-22 rose sharply in the milk of UI goats when infection was out of control. The results indicate that defenses mobilized by the mammary gland at an early stage of infection were essential to prevent staphylococci from reaching critical concentrations. Staphylococcal exotoxin production appeared to be a consequent event inducing the evolution to gangrenous mastitis.