Increased Serum TNF-alpha Research Articles

Air pollution influence on serum inflammatory interleukins: A prospective study in childhood-onset systemic lupus erythematous patients.

To assess the effect of individual exposure, in real-time, to traffic-related pollutants on serum interleukin levels of childhood-onset lupus erythematous systemic (c-SLE) patients. A longitudinal and observational design was conducted in 12 repeated measures of serum samples and clinical evaluations (totaling 108 measurements) of c-SLE patients over 30 consecutive months. Real-time, individual exposure to fine particles (PM2.5) and nitrogen dioxide (NO2) was measured with portable monitors. Generalized estimating equation was used to evaluate the association between exposure to PM2.5 and NO2 and the following serum cytokine levels on the 7days preceding clinical assessment and serum collection: MCP1, IL-6, IL-8, IL-10, IL-17, IFN-alpha, and TNF-alpha. Disease activity and other risk factors were also controlled. An interquartile range (IQR) increase in PM2.5 daily concentration was significantly associated with increased levels of TNF-alpha on the third, fourth, and seventh day after exposure; IL-10 on the third and fourth day after exposure; IL-17 on the third and seventh day after exposure; and INF-alpha on the third day after exposure (p < 0.05). An IQR increase in 7-day moving average of PM2.5 was associated with a 6.2pg/mL (95% CI: 0.5; 11.8; p = 0.04) increase in serum IFN-alpha level. An unexpected significant association was observed between an IQR increase in NO27-day cumulative concentration and a decrease of 1.6pg/mL (95% CI: -2.6; -0.7; p < 0.001) in serum IL-17. Real-time exposure to PM2.5 prospectively associated with increased serum TNF-alpha, INF-alpha, IL-10, and IL-17 levels in c-SLE patients.

M10. SERUM AND CLINICAL MARKERS ASSOCIATED WITH MEMBRANE PUFA CONTENT IN A POPULATION OF STABILISED AND TREATED SCHIZOPHRENIA OUTPATIENTS

BackgroundVarious candidate biomarkers have been proposed in schizophrenia to better characterise the several forms of the disorder. Among these candidates, peripheral (e.g. immunological, inflammatory, microbial) and brain (structural and functional) factors have been suggested and could help subtyping schizophrenia. One other potential biomarker, the membrane polyunsaturated fatty acid (PUFA) content, has been described both in peripheral cells and brain membranes of patients. Increased symptom severity along with more severe cognitive impairment were shown to be associated with decrease in the red blood cell (RBC) membrane sphingomyelin and PUFA content. However, little is known regarding the association between this membrane biomarker and serum related ones.The results presented here are preliminary and come from a study examining clinical and cognitive symptoms, RBC total membrane fatty-acid (FA) content, and serum biomarkers in a population of schizophrenia patients.We hypothesised that the sub-group of patients with decreased membrane PUFA content will also exhibit a higher incidence of abnormalities in some other serum biomarkers in comparison with patients with normal membrane PUFA content.MethodsA population of 33 chronic, medicated, and clinically stable patients with schizophrenia gave their informed consent for a study examining in particular their medical, psychiatric, and treatment history. The other measures assessed psychopathology (PANSS, CGI, Calgary scale), functioning and quality of life (GAF, SF36), and cognition (MOCA, MCST). Various biological biomarkers - in particular those related to inflammation, immunity, energy and hormone metabolism - were also examined. The total RBC membrane FA content was also measured using GC-FID (Gas Chromatography–Flame-Ionization Detection).ResultsBased on the standard values of the docosohexaenoic acid (DHA) (expressed as percentage of total fatty acids) in the RBC lipid membrane, a subgroup of 18 patients exhibited an abnormal decrease in DHA content (DHA-), while the remaining 15 individuals had DHA value within the normal range (DHAn). DHA- and DHAn groups were compared using univariate analysis in terms of other clinical and biological associated characteristics.We could also demonstrate a decrease in total membrane PUFAs in the DHA- group (p=0.004). Of interest, serum IL-6 was significantly (p=0.02) increased in the DHA- population. There was also a trend in favour of an increase in serum TNF-alpha in DHA- (mean value 30.5 vs 21 in DHA- and DHAn, respectively). In addition, the DHA- group have had significantly (p=0.03) more hospitalisations since the beginning of the disorder. The duration of untreated psychosis was 4 years vs 1 year in the DHA- vs DHAn groups, respectively. The same applied for the mean duration of hospitalisations (45 vs 34 days in DHA- vs DHAn). These values could not be explained by a longer duration in disease treatment in the 2 subpopulations.DiscussionThe presented data are in accordance with repeated findings showing decreased RBC membrane DHA and total PUFA content in a subgroup of individuals meeting criteria for schizophrenia. The splitting of the studied population into 2 groups numerically comparable as a function of their RBC membrane DHA content was thus justified. Interestingly, patients with lower membrane DHA content also exhibited abnormalities in other known biomarkers described in schizophrenia such as increased inflammation and disease severity. The presented data also show for the first time a biomarker associated with the duration of untreated psychosis. Altogether, these results, despite preliminary, are paving the way towards the identification of schizophrenia subtypes.

Animal Models of Steatosis (NAFLD) and Steatohepatitis (NASH) Exhibit Hepatic Lobe-Specific Gelatinases Activity and Oxidative Stress.

Animal models of obstructive cholestasis and ischemia/reperfusion damage have revealed the functional heterogeneity of liver lobes. This study evaluates this heterogeneity in nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH) rat models. Twelve-week-old Obese and Lean male Zucker rats were used for NAFLD. Eight-week-old male Wistar rats fed with 8-week methionine-choline-deficient (MCD) diet and relative control diet were used for NASH. Gelatinase (MMP-2; MMP-9) activity and protein levels, tissue inhibitors of metalloproteinase (TIMPs), reactive oxygen species (ROS), and thiobarbituric acid-reactive substances (TBARS) were evaluated in the left (LL), median (ML), and right liver (RL) lobes. Serum hepatic enzymes and TNF-alpha were assessed. An increase in gelatinase activity in the NASH model occurred in RL compared with ML. TIMP-1 and TIMP-2 displayed the same trend in RL as ML and LL. Control diet RL showed higher MMP-9 activity compared with ML and LL. No significant lobar differences in MMP-2 activity were detected in the NAFLD model. MMP-9 activity was not detectable in Zucker rats. TIMP-1 was lower in LL when compared with ML while no lobar differences were detectable for TIMP-2 in either Obese or Lean Zucker rats. Control diet rats exhibited higher ROS formation in LL versus RL. Significant increases in TBARS levels were observed in LL versus ML and RL in control and MCD rats. The same trend for ROS and TBARS was found in Obese and Lean Zucker rats. An increased serum TNF-alpha occurred in MCD rats. A lobar difference was detected for MMPs, TIMPs, ROS, and TBARS in both MCD and Zucker rats. Higher MMP activation in RL and higher oxidative stress in the LL, compared with the other lobes studied, supports growing evidence for functional heterogeneity among the liver lobes occurring certainly in both NAFLD and NASH rats.

P2.06-19 Targeting Polyamines as Potential Adjuvant Therapy in Malignant Pleural Mesothelioma Xenograft Models

Inhaling asbestos fibers is one of the commonest of malignant pleural mesothelioma (MPM). Although the import and use of asbestos have been restricted, the incidence of MPM is still rising due to a long lag time in malignant transformation. In 2004, the US Food and Drug Administration approved a combination of pemetrexed with cisplatin for treatment of unresectable MPM. At the same time, development of novel adjuvant therapeutic options for resected early-stage disease is also urgently needed. Ornithine decarboxylase (ODC) is highly expressed in 211H and H226 MPM xenografts and clinical tumor samples. Upregulation of ODC increases polyamine production and enhances tumor growth. a-difluoromethylornithine (DFMO) is a specific ODC inhibitor. Recent preclinical studies have demonstrated the adjuvant effect of DFMO in colon cancers using xenograft model. However, adjuvant effect of DFMO in MPM has not yet been studied. This study aims to disclose the adjuvant effect of DFMO in MPM xenograft models. The findings from this study will provide scientific foundation for future design of clinical trials of DFMO for adjuvant therapy in early disease for advanced MPM. Nude mice were fed with DFMO in drinking water 7 days before subcutaneous inoculation of 200,000 tumor cells (211H (biphasic) or H226 (epithelioid)). Mice with tumor size >600mm3 were considered reaching humane endpoint. Spermidine levels, protein expression, cytokines concentrations, NFkB translocation and apoptosis were investigated by Dot plot, Western blot, ELISA, immunofluorescence staining and TUNEL assay respectively. DFMO suppressed tumor growth in both xenografts. DFMO increased median survival from 49.5 days in control arm to 65 days in treatment arm in mice with 211H xenografts (p = 0.08), while from 44 days to 120 days in those with H226 xenografts (p = 0.0002). In H226 xenograft model, 43% of treated mice have not yet reached humane endpoint, mimicking long-term survival. Upon DFMO treatment, decrease in spermidine level, increase in nitrotyrosine content, nuclear translocation of NFkB, elevation of serum IL-6 and activation of apoptosis were observed in both xenografts. In addition, increase in nitrocysteine level, decrease in serum keratinocyte chemoattractant (KC), increase in serum TNF alpha, elevation of DNA lesion and inhibition of Akt/mTOR pathway were induced by DFMO in H226 xenografts, which may explain higher potency of DFMO in this xenograft. DFMO may have a potential role as adjuvant therapy in MPM especially epithelioid mesothelioma. Acknowledgment: This research was supported by Hong Kong Pneumoconiosis Compensation Fund Board, HKSAR.

Evaluation of Serum TNF-alpha, IL-6, IL-10, and IFN-gamma Levels in Patients with Crimean–Congo Hemorrhagic Fever

BackgroundCrimean–Congo hemorrhagic fever (CCHF) is a potentially fatal disease caused by a tick-borne virus from the Bunyaviridae family. Cytokines plays an important role in the pathogenesis of viral, bacterial, and immunologic diseases. This study aimed to investigate the role of TNF-alpha, IL-6, IL-10, and IFN-gamma levels in the severity of infection and clinical outcome of patients with CCHF.MethodsPatients with confirmed CCHF were divided into two groups (severe cases: Patients who exhibited hemorrhage during their hospital stay, and mild/moderate cases: Patients who displayed no hemorrhage during their hospital stay). Demographic characteristics, laboratory tests on admission of all patients with CCHF were investigated, and serum TNF-alpha, IL-6, IL-10, and IFN-gamma levels were measured.ResultsA total of 154 patients with confirmed CCHF were investigated. Forty-six (29.9%) of these patients were in the severe group. In patients with severe CCHF, significantly higher serum levels of TNF-alpha (68.2 ± 23.5; P = 0.008) and IL-6 (73.1 ± 41.6; P = 0.003) were detected, compared with cytokine levels in patients who mild/moderate CCHF (Table 1). No differences in serum IL-10 and IFN-gamma levels between patients who severe CCHF and those who mild/moderate CCHF were detected (P > 0.05).Table 1: Cytokine levels, demographic and laboratory characteristics in patients with severe and mild/moderate cases with CCHF.FeaturesSevere cases n = 46Mild/moderate cases n = 108 P-valueAge50.6 ± 20.349.8 ± 21.00.682Female gender, n (%)31 (67.4)63 (58,3)0.291 TNF- 68.2 ± 23.5 41.3 ± 17.4 0.008 IL-6 73.1 ± 41.6 38.0 ± 19.5 0.003 IL-106.32 ± 1.36.21 ± 1.40.753IFN-gamma145 ± 96126 ± 920.664WBC3286 ± 5,6022,275 ± 1,2860.280 PLT 53,564 ± 36,520 98,065 ± 42,768 0.001 CRP 3.2 ± 2.6 1.1 ± 1.4 0.005 ALT 521 ± 482 208 ± 320 0.044 AST 869 ± 1.182 256 ± 215 0.016 CPK 1,138 ± 970 676 ± 835 0.007 LDH 1,800 ± 1,254 589 ± 271 0.002 ConclusionCytokines, chemokines, and other inflammatory mediators function in a manner, acting on many different cell types to regulate the host’s immune response. When cytokines present in high concentrations, they might toxic or even lethal effects. In accordance with this view, we have detected increased serum TNF-alpha, IL-6 levels in the patients with severe CCHF.Disclosures All authors: No reported disclosures.

Efficacy and clinical observation of Shenling Baizhu Pill in treating irritable bowel syndrome

Objective To observe the clinic efficacy of Shenling Baizhu Pill on patients with irritable bowel syndrome(IBS). Methods Ninety patients with IBS were collected and divided into Chinese patent medicine group and western medicine group by random number method,45 patients in each group,the Chinese patent medicine group received Shenling Baizhu pill group treatment,the western medicine group was gave Smectite Powder treatment.The changes of clinical symptoms,intestinal tract microorganism and serum inflammatory factor of patients were observed and compared between two groups before and after treatment.The clinical symptoms was evaluated by questionnaire,the Lactobacillus,Bifidobacterium,enterobacteria and yeast were determined by cultivation,the levels of IL-10,IL-18 and TNF-αwere detected by enzyme linked immunosorbent assay (ELISA). Results Two groups before treatment of anaerobic bacteria in feces reduction,aerobic bacteria increased serum IL-18 and TNF-alpha content,increased,the content of IL-10 decreased.Shenling Baizhu Pill intervention for 8 weeks in patients with intestinal anaerobic bacteria[(B/E 1.07±0.49)]and serum IL-10 [(43.65±9.52)mg/L]content increased,aerobic bacteria[(B/E 1.07±0.49)],IL-18[(82.06±21.07)mg/L] and TNF-alpha[(28.44±10.20)mg/L]decreased.The IL-18 decreased the most,was different with the western medicine group,there was a statistical significance(t=9.926,P<0.01).Two groups had no bad reaction. Conclusion Shenling Baizhu Pill can effectively regulates and improves the intestinal micro-environment,promote the growth of intestinal probiotics and remodeling the intestinal function of biological barrier, decrease the levels of proinflammatory factors and rise the anti-inflammatory factors level,which has a good clinic effect in treating IBS. Key words: Shenling Baizhu Pill; Irritable bowel syndrome; Intestinal microecology

Increased Serum TNF-Alpha and Matrix Metalloproteinase-2 are Associated With Grip Strength Declines and Tissue Degeneration in a Rat Model of Overuse

We have shown that continued performance of repetitive tasks induces grip strength declines despite resolution of systemic inflammation. We hypothesize this is due to underlying tissue degeneration. Here, we assessed long term performance (18 weeks) of a high-repetition, low-force (HRLF) task in a rat model of reaching and grasping. We observed reduced grip strength immediately after training, and persistent grip strength declines in reach limbs of HRLF rats. Several inflammatory cytokines increased in serum of 6- and 12-week HRLF rats, e.g. tumor necrosis factor alpha (TNF-alpha). TNF- alpha was also increased in reach limb muscles and tendons at similar time points. A serum analyte of collagen degradation (matrix metalloproteinase-2, MMP2) was increased in serum of 18-week HRLF rats. MMP2 and several other MMPs, as well as two fibrogenic proteins (CTGF and TGFB1), were increased in 18-week HRLF tendons, which also showed histological signs of pathology. Thus, motor declines were associated earlier with tissue inflammation but later with tendon degenerative changes. Assaying for TNF- alpha and MMP2 provided important insights into the stages of inflammation and degradation in this model.

Takayasu's Arteritis in a Patient with Crohn's Disease: An Unexpected Association during Infliximab Therapy

We report a 20-year-old woman with Crohn's disease (CD) who developed anterior neck pain while being treated with the anti-tumor necrosis factor (TNF)-alpha monoclonal antibody, infliximab. She showed no symptoms suggestive of active CD except for tenderness along the left common carotid artery with marked increases in serum TNF-alpha and inflammatory reactions. Based on thickened walls of large vessels with enhancement effects on computed tomography, she was diagnosed as having associated Takayasu's arteritis (TA), which was successfully treated with corticosteroid. Even if CD is controlled by infliximab, other autoimmune disorders, such as TA, may develop as a complication.

The effect of probiotic microorganisms and bioactive compounds on chemically induced carcinogenesis in rats

Diet interventions and natural bioactive supplements have now been extensively studied to reduce risks of colon cancer, which is one of the major public health problem throughout the world. The objective of our investigation was to study the effects of probiotic, prebiotic, nutritional plant extract, and plant oil on selected biochemical and immunological parameters in rats with colon cancer induced by N,N dimethylhydrazine (DMH). Male and female Wistar albino rats were were fed by a high-fat (HF) diet (10% fat in the diet) and were divided into 9 groups: Control group; PRO group - HF diet supplemented with probiotic Lactobacillus plantarum to provide 3 x 109 c.f.u. of strain/1 ml of medium; PRE group - HF diet supplemented with inulin enriched with oligofructose (2% of HF diet); HES group - HF diet supplemented with plant extract of Aesculus hippocastanum L. (1% of HF diet); OIL group - HF diet comprised Linioleum virginale (2% of HF diet); and combination of probiotic microorganisms and bioactive compounds in the groups - PRO-PRE, PRO-HES, PRO-OIL, PRE-OIL. Carcinogenesis was initiated with subcutaneous injection of DMH (20 mg/kg) two times at week interval and dietary treatments were continued for the six weeks. Application of probiotic microorganisms and bioactive compounds in all treated groups significantly decreased the activities of bacterial enzymes (p<0.001), the fecal bile acids concentration (p<0.01; p<0.001) and significantly increased serum TNFalpha level (p<0.001) in comparison to the control rats. The number of coliforms was reduced in PRO, PRO-PRE, PRO-OIL and PRE-OIL groups and significantly higher count of lactobacilli (p<0.05) was observed in PRO-PRE, PRO-OIL and PRE-OIL groups in compare with the controls. In conclusion, the results of this study indicate that probiotic microorganisms and bioactive compounds could exert a preventive effect on colon carcinogenesis induced by DMH.

Tumor necrosis factor alpha gene variants do not display allelic imbalance in circulating myeloid cells

Carriage of the TNF -308 A allele (rs1800629 A) has been associated with increased serum TNF-alpha levels, the development of sepsis syndrome, and fatal outcome, in severely traumatized patients (Menges et al., 2008 [1]). Herein, we analysed the putative allelic imbalance of TNF-alpha release from myeloid cells. Circulating peripheral blood cells from healthy human blood donors (n=104) and monocyte-derived macrophages (n=158) were analysed for their ex vivo capacity of TNF-alpha expression. Our findings indicate that carriage of the TNF -308 A allele is not associated with high TNF-alpha expression in circulating human leucocytes and monocyte-derived macrophages. Other cellular sources, e.g. tissue-resident cells like mast cells and/or tissue specific macrophages might be the cellular source of high TNF-alpha serum levels shortly after trauma.

Lipopolysaccharide exposure is linked to activation of the acute phase response and growth failure in pediatric Crohnʼs disease and murine colitis

Systemic exposure to lipopolysaccharide (LPS) has been linked to clinical disease activity in adults with inflammatory bowel disease (IBD). We hypothesized that markers of LPS exposure and the acute phase response (APR) would be increased in pediatric IBD patients with growth failure, and that LPS signaling would be required for induction of the APR in murine colitis. Serum markers of LPS exposure, endotoxin core IgA antibody (EndoCAb), and the APR, LPS binding protein (LBP) were quantified in pediatric IBD patients and controls. LBP and cytokine production were determined after administration of trinitrobenzene sulfonic acid (TNBS) enemas to mice with genetic deletion of Toll-Like receptor 4 (TLR4), and wildtype (WT) controls. Serum EndoCAb and LBP were significantly elevated in patients with Crohn's disease (CD), compared to disease controls with ulcerative colitis (UC) and healthy controls (P < 0.001). This was independent of disease activity or location. CD patients with elevated serum EndoCAb and LBP exhibited linear growth failure which persisted during therapy. Serum LBP increased in WT mice following TNBS administration, in conjunction with increased serum TNF-alpha, IL-6, and IL-10, and expansion of regulatory T-cell numbers. Both the APR and expansion of foxp3+ T cells were abrogated in TLR4-deficient mice, in conjunction with a reduction in acute weight loss. LPS exposure and a persistent APR are associated with growth failure in pediatric CD. LPS signaling is required for the APR in murine colitis. Therapies targeting this pathway may benefit the subset of patients with refractory growth failure.

TNF-α modulates iNOS expression in an experimental rat model of indomethacin-induced jejunoileitis

Multiple mucosal immune factors, such as TNF-alpha and IL-1beta, are thought to be key mediators involved in inflammatory bowel disease. We evaluated the role of the pro-inflammatory cytokine TNF-alpha on nitric oxide synthase (NOS) expression in indomethacin-induced jejunoileitis in rats. Jejunoileitis was induced in rats with subcutaneous injections of indomethacin (7.5 mg/kg) 24 h apart for two consecutive days, and animals were randomized into four groups. Group 1 received only indomethacin. Group 2 was treated with a daily dose of phosphodiesterase (PDE) inhibitor (theophylline or pentoxifylline) by oral gavage for 2 days before and 4 days after indomethacin. Group 3 received a single dose of anti-TNF-alpha monoclonal antibody (TNF-Ab, IP) 30 min before indomethacin. Group 4 was treated with 1 h hyperbaric oxygenation (HBO(2)) for 5 days after indomethacin. Rats were sacrificed at 12 h or 4 days after final indomethacin injection. PDE inhibitor, TNF-Ab, or HBO(2) treatment significantly decreased indomethacin-induced ulceration, myeloperoxidase activity, and disease activity index. Although indomethacin significantly increased serum TNF-alpha and nitrate/nitrite (NOx) concentrations above control values at 12 h, inducible NOS (iNOS) expression was detected only at day 4. Serum IL-1beta levels did not change at 12 h but increased 4-fold after 4 days. Indomethacin had no effect on constitutive NOS. Treatment with PDE inhibitor, TNF-Ab, or HBO(2) significantly reduced serum/tissue TNF-alpha, IL-1beta, NOx, and iNOS expression. Our data show TNF-alpha plays an early pro-inflammatory role in indomethacin-induced jejunoileitis. Additionally, down-regulation of NOx by PDE inhibitors, TNF-Ab, or HBO(2) suggests that TNF-alpha modulates iNOS expression.

Effects of probiotic therapy on portal pressure in patients with cirrhosis: a pilot study

Recent literature has supported the role of bacterial translocation as a mediator of splanchnic vasodilatation and portal hypertension. The objective of this study was to determine whether the probiotic VSL#3 would reduce portal pressure in patients with cirrhosis. Eight patients with compensated or very early decompensated cirrhosis and hepatic venous pressure gradient (HVPG) >10 mmHg, received 2 months of VSL#3 (3600 billion bacteria daily). The HVPG, intestinal permeability, endotoxin, tumour necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-8, renin and aldosterone were measured at baseline and study end. There was no change in the HVPG or intestinal permeability from baseline to study end but there was a trend to reduction in plasma endotoxin (P=0.09), a mild but significant increase in serum TNF-alpha (P=0.02) and a significant reduction in plasma aldosterone (P=0.03). Within the limitations of small sample size, there does not appear to be a benefit of probiotic therapy for portal pressure reduction in patients with compensated or early decompensated cirrhosis. The reductions in endotoxin and aldosterone suggest possible beneficial effects of probiotics for this patient population. The clinical significance of the small but unexpected increase in TNF-alpha is unclear. Future studies are planned in patients with decompensated cirrhosis.

Tetrandrine Increased the Survival Rate of Mice With Lipopolysaccharide-Induced Endotoxemia

Endotoxemia usually causes significant morbidity and mortality, and treatment of endotoxemia is often ineffective. The effects of tetrandrine (a bisbenzylisoquinoline alkaloid) on lipopolysaccharide (LPS)-induced endotoxemia were investigated in mice. The peritoneal macrophages were stimulated with LPS and treated with or without tetrandrine. The amounts of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, IL-6, and IL-10 secreted by peritoneal macrophages were measured by enzyme-linked immunosorbent assay. Mice were intraperitoneally injected with LPS to induce endotoxemia and were treated or not treated with oral gavage with 150 mg/kg tetrandrine 1 hour before or after LPS injection. The survival rate of the mice was determined after they were treated with various regiments. The amounts of TNF-alpha, IL-1beta, IL-6, and IL-10 in the serum of the mice were measured by enzyme-linked immunosorbent assay, and the high mobility group box 1 (HMGB1) concentration was studied by Western blot analysis. Tetrandrine suppressed the LPS-induced increase of TNF-alpha, IL-1beta, and HMGB1 secretion by peritoneal macrophages but did not affect the IL-6 and IL-10 concentrations. The animals treated with tetrandrine either 1 hour before or after LPS injection had a 100% survival rate, which was significantly higher than that of the control group (40%) (p = 0.005). The LPS-induced increase in serum TNF-alpha, IL-1beta, and HMGB1 concentrations was reduced by tetrandrine treatment administered either 1 hour before or after LPS injection (p < 0.0001). In contrast, tetrandrine prolonged the LPS-induced elevation in serum IL-10 concentrations only mildly changed the serum IL-6 concentrations. Tetrandrine treatment either 1 hour before or 1 hour after LPS injection reduced the mortality rate of the mice with LPS-induced endotoxemia. The effects of tetrandrine on LPS-induced endotoxemia might be related to the suppression of TNF-alpha, IL-1beta, and HMGB1 concentrations.

Effect of Ocimum sanctum, ascorbic acid, and verapamil on macrophage function and oxidative stress in mice exposed to cocaine

Objective:To investigate the effect of Ocimum sanctum, ascorbic acid, and verapamil on macrophage function and oxidative stress in experimental animals exposed to cocaine.Materials and Methods:Mice were used in this study and were divided randomly into different groups of six animals each. They were either treated with intraperitoneal injection of saline or cocaine hydrochloride or an oral feeding of oil of Ocimum sanctum, ascorbic acid or verapamil, or both (ascorbic acid and verapamil), and were evaluated for a respiratory burst of macrophages, superoxide and nitric oxide (NO) production, estimation of TNF-α in the serum and supernatant of cultured macrophages, estimation of lipid peroxidation (malondialdehyde- MDA) in the serum, and superoxide dismutase activity in the erythrocytes.Results:Unstimulated respiratory burst as well as superoxide production was enhanced on treatment with cocaine and all the three drugs were found to attenuate this enhancement. The bactericidal capacity of macrophages decreased significantly on chronic cocaine exposure, as it was associated with decreased respiratory burst and superoxide production. There was a significant decrease in NO production by macrophages on chronic cocaine exposure and all the test drugs were found to restore nitrite formation to a normal level. There was an increase in the malonylodialdehyde (MDA) level and decrease in the superoxide dismutase level on chronic cocaine exposure, and all the three drugs effectively decreased the MDA level and increased superoxide dismutase level. There was an increase in serum TNF-α on chronic cocaine exposure, which was decreased significantly by ascorbic acid and verapamil.Conclusion:O. sanctum, ascorbic acid, and verapamil were equally effective in improving the macrophage function and reducing oxidative stress. These findings suggested that O. sanctum, ascorbic acid, and verapamil attenuated acute and chronic cocaine-mediated effects.

Metallothionein attenuates carmustine-induced oxidative stress and protects against pulmonary fibrosis in rats

The present study was carried out to evaluate the effect of exogenously administered metallothionein (MT) against carmustine (BCNU)-induced lung toxicity in rats. A total of 60 rats were randomly divided into four groups (15/group): control group in which the animals received 0.5 ml physiologic saline containing 10% ethanol (IP) weekly, MT-administered group in which rats received MT (30 micromol/kg, IP) weekly, BCNU-administered group in which rats received BCNU (5 mg/kg, IP) weekly and MT + BCNU group in which rats received weekly doses of BCNU (5 mg/kg, IP) followed 24 h later by MT (30 micromol/kg, IP). At the end of the experiment (after 6 weeks), lung histological changes, collagen staining, the activity of glutathione reductase (GR) and contents of reduced glutathione (GSH) and hydroxyproline (Hpr) in the lung as well as serum level of tumor necrosis factor-alpha (TNF-alpha) were evaluated. The obtained data revealed that BCNU induced pathological changes and markedly increased lung collagen and level of Hpr but decreased GSH content and GR activity and increased serum TNF-alpha compared to both control and MT-administered rats. Administration of MT + BCNU markedly improved histological features and decreased staining of collagen along with increased GR activity, GSH content but decreased level of Hpr in lung tissue as well as decreased serum level of TNF-alpha compared with BCNU-treated rats. Based on our results, it is possible to postulate that exogenous MT can act against BCNU-induced lung toxicity by a mechanism related, at least in part, to its ability to decrease oxidative stress and fibrosis.

RU486 did not exacerbate cytokine release in mice challenged with LPS nor in db/db mice

BackgroundGlucocorticoids down-regulate cytokine synthesis and suppress inflammatory responses. The glucocorticoid receptor (GR) antagonist RU486 may exacerbate the inflammatory response, and concerns over this exacerbation have limited the development and clinical use of GR antagonists in the treatment of diabetes and depression. We investigated the effects of RU486 on serum cytokines in db/db mice and on lipopolysaccharide (LPS)-induced circulating TNFα levels in both normal AKR mice and diet-induced obese (DIO) C57BL/6 mice.ResultsChronic treatment of db/db mice with RU486 dose-dependently decreased blood glucose, increased serum corticosterone and ACTH, but did not affect serum MCP-1 and IL-6 levels. LPS dose-dependently increased serum TNFα in both AKR and C57BL/6 DIO mice, along with increased circulating corticosterone and ACTH. Pretreatment of the mice with RU486 dose-dependently suppressed the LPS induced increases in serum TNFα and further increased serum corticosterone.ConclusionRU486 at doses that were efficacious in lowering blood glucose did not exacerbate cytokine release in these three mouse models. RU486 actually suppressed the lower dose LPS-mediated TNFα release, possibly due to the increased release of glucocorticoids.

Autoantibodies to the translational suppressors T cell intracytoplasmic antigen 1 and T cell intracytoplasmic antigen 1–related protein in patients with rheumatic diseases: Increased prevalence in systemic lupus erythematosus and systemic sclerosis and correlation with clinical features

T cell intracytoplasmic antigen 1 (TIA-1) and TIA-1-related protein (TIAR) are involved in posttranscriptional regulation of the expression of tumor necrosis factor alpha (TNFalpha) and other proteins. Given the pivotal role of TNFalpha in chronic inflammatory diseases, this study was undertaken to analyze sera from patients with systemic autoimmune diseases for the presence of autoantibodies to TIA proteins and to investigate the expression of these proteins in inflamed tissue. The presence of autoantibodies to TIA proteins in sera from 385 patients with rheumatic diseases and healthy controls was determined by immunoblotting using recombinant antigens. Expression of TIA proteins in skin and kidney tissue was analyzed by immunohistochemistry. Serum levels of TNFalpha were measured by enzyme-linked immunosorbent assay. Autoantibodies to TIA-1 and/or TIAR were detected in 61% of patients with systemic lupus erythematosus (SLE), 42% of patients with systemic sclerosis (SSc), 15-31% of patients with other rheumatic diseases, and 6% of healthy controls. Compared with patients negative for anti-TIA antibody, anti-TIA antibody-positive SLE patients had higher disease activity (P = 0.01), elevated antibodies to double-stranded DNA (P = 0.0003), and increased serum TNFalpha levels (P = 0.018). In SLE patients, anti-TIAR antibodies were associated with lupus nephritis (P = 0.02), while in patients with SSc, anti-TIA-1 was associated with lung involvement (P = 0.02). Immunohistochemical analysis of skin and kidney tissue revealed aberrant expression of TIA proteins in skin lesions from SLE and SSc patients, as well as in glomerular cells from SLE patients. Aberrant expression of TIA proteins in inflammatory tissue may lead to systemic autoantibody responses, particularly in SLE and SSc. Increased occurrence of anti-TIA autoantibodies in patients with severe organ involvement may point to a possible pathogenetic role.

Deletion of a single mevalonate kinase (Mvk) allele yields a murine model of hyper‐IgD syndrome

In the current study our objective was to develop a murine model of human hyper-IgD syndrome (HIDS) and severe mevalonic aciduria (MA), autoinflammatory disorders associated with mevalonate kinase deficiency (MKD). Deletion of one Mvk allele (Mvk (+/-)) yielded viable mice with significantly reduced liver Mvk enzyme activity; multiple matings failed to produce Mvk (-/-) mice. Cholesterol levels in tissues and blood, and isoprene end-products (ubiquinone, dolichol) in tissues were normal in Mvk (+/-) mice; conversely, mevalonate concentrations were increased in spleen, heart, and kidney yet normal in brain and liver. While the trend was for higher IgA levels in Mvk (+/-) sera, IgD levels were significantly increased (9-12-fold) in comparison to Mvk (+/+) littermates, in both young (<15 weeks) and older (>15 weeks) mice. Mvk (+/-) animals manifested increased serum TNF-alpha as compared to wild-type littermates, but due to wide variation in levels between individual Mvk (+/-) mice the difference in means was not statistically significant. Mvk (+/-) mice represent the first animal model of HIDS, and should prove useful for examining pathophysiology associated with this disorder.

Silymarin, the Antioxidant Component of Silybum marianum, Prevents Sepsis-Induced Acute Lung and Brain Injury

Sepsis is associated with enhanced generation of reactive oxygen species, which leads to multiple organ dysfunctions. Based on the potent antioxidant effects of silymarin, we investigated the putative protective role of silymarin against sepsis-induced oxidative damage in lung and brain tissues. Sepsis was induced by cecal ligation and perforation (CLP). Sham and CLP groups received either vehicle or silymarin (50 mg/kg, p.o.) or 150 mg/kg i.p. N-acetylcysteine (NAC) for 10 days prior and immediately after the operation. Six hours after the surgery, rats were decapitated and blood was collected for the measurement of proinflammatory cytokines (tumor necrosis factor-alpha, interleukin-1 beta [IL-1 beta], and IL-6) levels, lactate dehydrogenase activity, and total antioxidant capacity. Lung and brain samples were taken for the measurement of malondialdehyde and glutathione levels, myeloperoxidase activity, thromboplastic activity, and also for histological assessment. Formation of reactive oxygen species in tissue samples was monitored by using chemiluminescence technique with luminol and lusigenin probe. Sepsis increased serum TNF-alpha, IL-1 beta, IL-6 levels, and lactate dehydrogenase activity and decreased total antioxidant capacity. On the other hand, tissue glutathione levels were decreased while malondialdehyde levels and myeloperoxidase activity were increased in both the lung and the brain tissues due to CLP. Furthermore, luminol and lucigenin chemiluminescence were significantly increased in the CLP group, indicating the presence of the oxidative damage. Silymarine and NAC treatment reversed these biochemical parameters and preserved tissue morphology as evidenced by histological evaluation. Silymarin, like NAC, reduced sepsis-induced remote organ injury, at least in part, through its ability to balance oxidant-antioxidant status, to inhibit neutrophil infiltration, and to regulate the release of inflammatory mediators.