Identification of novel blood biomarker panels to detect ischemic stroke in patients and their responsiveness to therapeutic intervention.

The use of blood biomarkers for stroke has been long considered an excellent method to determine the occurrence, timing, subtype, and severity of stroke. In this study, venous blood was obtained from ischemic stroke patients after stroke onset and compared with age and sex-matched controls. We used a multiplex panel of 37 inflammatory molecules, analyzed using Luminex MagPix technology, to identify the changes in plasma proteins after ischemic stroke. We identified eight key molecules that were altered within the blood of stroke patients as compared to controls. Plasma levels of interleukin 6 signal transducer (sIL-6Rß/gp130), matrix metalloproteinase-2 (MMP-2), osteopontin, sTNF-R1 and sTNF-R2 were significantly higher in stroke patients compared to controls. Interferon-ß, interleukin-28, and thymic stromal lymphopoietin (TSLP) were decreased in plasma from stroke patients. No other immunological markers were significantly different between patient groups. When stroke patients were treated with tissue plasminogen activator (t-PA), plasma levels of interferon-α2 significantly increased while interleukin-2 and pentraxin-3 decreased. The discriminatory power of the molecules was evaluated by receiver operating characteristic (ROC) analysis. According to ROC analysis, the best markers for distinguishing stroke occurrence were MMP-2 (AUC = 0.76, sensitivity 62.5%, specificity 88.5%), sTNF-R2 (AUC = 0.75, sensitivity 83.3%, specificity 65.3%) and TSLP (AUC = 0.81, sensitivity 66.7%, specificity 96.2%). Multivariate logistic regression, used to evaluate the combination of proteins, identified a biomarker panel with high specificity and sensitivity (AUC = 0.96, sensitivity 87.5%, specificity 96.2%). These results indicate a novel set of blood biomarkers that could be used in a panel to identify stroke patients and their responsiveness to therapeutic intervention.

Curcumin Attenuates Inflammation in a Severe Acute Pancreatitis Animal Model by Regulating TRAF1/ASK1 Signaling.

BACKGROUND Inflammation plays an important role in initiation and development of severe acute pancreatitis (SAP). Curcumin exerts potent anti-inflammatory effects in many diseases, including acute pancreatitis. However, the specific molecular mechanisms are not clear. MATERIAL AND METHODS Intra-biliopancreatic duct injection of taurocholate was used to establish an animal model of SAP. Curcumin was administrated to animals as pre-treatments. Concentrations of cytokines in serum and ascites were measured by enzyme-linked immunosorbent assay (ELISA). A colorimetric method was used to determine the amylase activity. Western blotting was used to examine the expression levels and phosphorylation levels of proteins. Immunoprecipitation was used to assess the molecular association between apoptosis signal- regulating kinase 1 (ASK1) and thioredoxin (Trx). RESULTS Pre-treatment with curcumin reduced the concentrations of interleukin (IL6) and tumor necrosis factor (TNFα) in serum and ascites, as well as the ascites volume and amylase activity in SAP rats. Pre-treatment with curcumin reduced the expression level of TNF receptor-associated factor 1 (TRAF1), IL6, and TNFa in pancreas in SAP rats. Moreover, the phosphorylation levels of mitogen-activated protein kinase (MAPK) kinase 4 (MKK4), MKK7, and c-Jun NH(2)-terminal protein kinase (JNK) were reduced by curcumin pre-treatment. The molecular association between ASK1 and Trx was recovered by curcumin pre-treatment. As a result, the nuclear translocation of nuclear factor kappa B (NF-κB) was suppressed in pancreases from SAP rats. CONCLUSIONS Activation of the TRAF1/ASK1/JNK/NF-κB signaling pathway is involved in the inflammation of SAP. Curcumin exerts anti-inflammatory effects by suppressing this proinflammatory pathway.

Increased serum levels of tumor necrosis factor receptor-associated factor 1 (TRAF1) correlate with disease activity and autoantibodies in rheumatoid arthritis.

BACKGROUND: In 2007 a genome wide association analyses revealed an additional risk-associated genetic region in rheumatoid arthritis (RA), the tumor necrosis factor receptor-associated factor 1-complement 5 (TRAF1-C5) containing locus on chromosome 9, comprehensive evaluation of the TRAF1 in RA patients remains necessary. METHODS: Serum was obtained from 79 RA patients and from 38 healthy individuals. Concentrations of TRAF1 were measured by enzyme-linked immune sorbent assay (ELISA). RESULTS: We found that the serum concentration of TRAF1 in RA patients was 35.9±51.2pg/ml, the serum concentration of TRAF1 in healthy controls was 12.5±8.6pg/ml. The difference between these 2 groups was significant (p=0.003). Patients with high disease activity had significantly higher TRAF1 concentration in comparison to patients with low and moderate disease activity (p<0.05). We also found that the numerical DAS28 had a significant positive correlation with TRAF1 concentration (r=0.419, p<0.001). We found that serum GPI and RF concentrations showed a significant positive correlation with TRAF1 concentrations respectively (r=0.767, p<0.001; r=0.365, p=0.001), while CCP concentration showed no significant correlation. CONCLUSIONS: TRAF1 plays a crucial role in the pathogenesis of autoantibodies and may serve as a serologic inflammatory marker of disease activity in RA patients.

[The prognostic role of clinical factors, VEGF, IL-8 and sTNF-R1 in cutaneous melanomas at locoregional stage]. / Znaczenie prognostyczne czynników klinicznych oraz oznaczania stezenia VEGF, IL-8 i sTNF-R1 w czerniakach skóry o zaawansowaniu lokoregionalnym.

UNLABELLED: The treatment results of patients with locoregional melanoma are still not satisfactory - up to 50% of patients experience recurrence and (or) disease dissemination. The aim of the study was to assess the prognostic value of clinical factors and serum cytokines of patients with cutaneous melanoma in locoregional stage. MATERIAL AND METHODS: 149 patients at stage I-III according AJCC treated between 2007-2010 were included. Pre-surgery serum levels of VEGF IL-8 and sTNF-R1 were analyzed by ELISA method in 74 melanoma patients and 50 healthy controls. The median follow-up time was 16 months (range: 1-81 months). RESULTS: The most important factors influencing the disease-free survival (DFS) are: staging system according to AJCC) (p < 0.001), regional nodal stage (pN) (p < 0.001), primary tumor (Breslow) thickness (pT) (p = 0.013) and melanoma ulceration (p = 0.004). The serum levels of selected cytokines were significantly higher in melanoma patients than in healthy volunteers (VEGF, p < 0.001; sTNF-R1, p < 0.001; IL-8, p = 0.001). There were no significant relationships between level of cytokine, recurrence or clinical/pathological parameters. CONCLUSIONS: The AJCC staging system gives the most accurate insight into prognosis of melanoma patients at locoregional stage after primary therapy. Cytokine serum profile in melanoma patients at locoregional stage has limited value for predicting tumor burden and treatment outcomes.

Levels of circulating interleukin-1 receptor antagonist and C-reactive protein in long-term survivors of testicular cancer with chronic cancer-related fatigue.

Low-grade inflammatory responses may be related to the pathogenesis of cancer-related fatigue (CRF). We investigated circulating levels of various inflammatory markers in relation to chronic CRF (6 month duration) in Norwegian long-term survivors of testicular cancer (TCSs). We compared 92 TCSs with chronic CRF (cases) to 191 TCS without (controls) at median age 45 years (range 23-73), and median 11 years post-treatment (range 5-20). Chronic CRF was defined using the Fatigue Questionnaire, while plasma concentrations of cytokines and serum CRP were determined by various immunoassays. Higher levels of interleukin-1 receptor antagonist (IL-1ra) (p=.002) and C-Reactive protein (CRP) (p=.036) were found in cases compared to controls. No differences were observed for interleukin-6 (IL-6), soluble Tumor Necrosis Factor Receptor type 1 (sTNF-R1) or neopterin. Both IL-1ra and CRP were correlated with physical but not with mental fatigue. In logistic regression analyses IL-1ra and CRP explained 3.5% and 2.8%, respectively, of the variance in chronic CRF. Single adjustments for depression, anxiety and neuroticism each raised the models' explained variance to approximately 35%. Those factors did not significantly alter the relationship between chronic CRF and IL-1ra/CRP. BMI and smoking emerged as possible confounding factors. These results indicate that chronic CRF in TCSs is associated with higher levels of circulating IL-1ra and CRP, possibly mediated by physiological morbidity. Hence, the findings lend some support to the hypothesis that low-grade inflammatory processes are involved in the pathogenesis of chronic CRF in cancer survivors.

The roles of TNF-alpha and the soluble TNF receptor I on sleep architecture in OSA.

OBJECTIVE: Patients with obstructive sleep apnea (OSA) have been described to have increased levels of inflammatory cytokines (particularly TNF-alpha) and have severely disturbed sleep architecture. Serum inflammatory markers, even in normal individuals, have been associated with abnormal sleep architecture. Not much is known about the role the TNF receptor plays in the inflammation of OSA nor if it is associated with changes in sleep architecture or arousals during the night. We hypothesized that the TNF receptor might play an important role in the inflammation as well as sleep architecture changes in patients with OSA. DESIGN: Thirty-six patients with diagnosed (AHI > 15) but untreated OSA were enrolled in this study. Baseline polysomnograms as well as TNF-alpha and soluble TNF receptor I (sTNF-RI) serum levels were obtained on all patients. We evaluated the association between serum levels of TNF-alpha and sTNF-RI with various polysomongraphic characteristics, including sleep stages and EEG arousals. RESULTS: sTNF-RI levels were significantly correlated with snore arousals (r value 0.449, p value 0.009), spontaneous movement arousals (r value 0.378, p value 0.025), and periodic limb movement arousals (r value 0.460, p value 0.008). No statistically significant correlations were observed with TNF-alpha to any polysomnographic variables. To control for statistical significance with multiple comparisons, a MANOVA was performed with TNF-alpha and sTNF-RI as dependent variables and sleep architecture measures and arousals as independent variables. The model for sTNF-RI was statistically significant (F value 2.604, p value 0.03), whereas the model for TNF-alpha was not, suggesting sleep quality significantly affects sTNF-RI. Hierarchal linear regression analysis demonstrated that sTNF-RI was independently associated with spontaneous movement arousal index scores after controlling for age, body mass index, and sleep apnea severity. CONCLUSIONS: These findings suggest that sTNF-RI is associated with arousals during sleep, but not with other measures in patients with OSA.

Changes of cytokines in cirrhosis patients with advanced hepatocellular carcinoma treated by intra-arterial chemotherapy.

INTRODUCTION: Tumor necrosis factor (TNF) induces cancer cell-specific apoptosis by binding to a TNF-related apoptosis-inducing ligand. Binding of the Fas ligand on cytotoxic T lymphocytes to the Fas receptor on hepatocytes is also known to induce apoptosis. The aim of this study was to clarify changes of cytokines in patients with liver cirrhosis (LC) and advanced hepatocellular carcinoma (aHCC) receiving intra-arterial combination chemotherapy. METHODS: Twenty-one adult Japanese LC patients with aHCC received intra-arterial combination chemotherapy. The serum levels of TNF-alpha, soluble TNF receptor-I (sTNFr-I), soluble Fas ligand (sFas L), and soluble Fas (sFas) were evaluated. RESULTS: Thirteen of the 21 patients (group R) showed an objective response, while the other eight patients (group N) showed no response. The serum level of TNF-alpha was lower after chemotherapy than before chemotherapy in group N, but there was no difference of serum sTNFr-I levels between before and after chemotherapy and there were also no differences between the two groups. The serum sFas levels were higher after chemotherapy than before chemotherapy in group N, while there was no difference among groups. CONCLUSIONS: These results indicate that a high serum TNF-alpha level and a low serum sFas level might be important for successful combined arterial chemotherapy in LC patients with aHCC.

Imbalance between soluble tumour necrosis factor receptors type 1 and 2 in chronic periodontitis.

BACKGROUND: Soluble types of tumour necrosis factor (TNF) receptors type 1 and 2 modulate the TNF-alpha-mediated inflammatory responses in chronic periodontitis (CP). OBJECTIVES: This study investigated the levels of TNF-alpha, soluble TNF receptor type 1 and 2 in gingival crevicular fluid (GCF) and serum of healthy subjects and CP patients. MATERIALS AND METHODS: Thirty-eight sera and 73 GCF samples were collected from 16 healthy subjects and 22 CP patients. GCF was collected from probing pocket depth (PPD)< or =3 mm sites of healthy subjects, PPD< or =3, 4-6 and > or =7 mm sites of CP patients. The levels of TNF-alpha, soluble TNF receptor type 1 and 2 in the serum and GCF were quantified by enzyme-linked immunosorbant assay. RESULTS: The total amounts of TNF-alpha, soluble TNF receptor type 1 and 2 in GCF significantly elevated with increasing PPD in both site-based (p<0.05) and subject-based (p<0.05) analyses. However, their levels progressively diverged as the pocket depths increased, with the soluble TNF receptor type 2 level being comparatively lower than type 1. On the other hand, soluble TNF receptor type 2/type 1 ratios in GCF decreased as the severity of periodontitis increased (p<0.0001). CONCLUSION: The imbalance between soluble TNF receptor type 1 and 2 levels in GCF could be related to CP severity.