Capillary electrophoresis mass spectrometry-based saliva metabolomics identified oral, breast and pancreatic cancer-specific profiles

Saliva is a readily accessible and informative biofluid, making it ideal for the early detection of a wide range of diseases including cardiovascular, renal, and autoimmune diseases, viral and bacterial infections and, importantly, cancers. Saliva-based diagnostics, particularly those based on metabolomics technology, are emerging and offer a promising clinical strategy, characterizing the association between salivary analytes and a particular disease. Here, we conducted a comprehensive metabolite analysis of saliva samples obtained from 215 individuals (69 oral, 18 pancreatic and 30 breast cancer patients, 11 periodontal disease patients and 87 healthy controls) using capillary electrophoresis time-of-flight mass spectrometry (CE-TOF-MS). We identified 57 principal metabolites that can be used to accurately predict the probability of being affected by each individual disease. Although small but significant correlations were found between the known patient characteristics and the quantified metabolites, the profiles manifested relatively higher concentrations of most of the metabolites detected in all three cancers in comparison with those in people with periodontal disease and control subjects. This suggests that cancer-specific signatures are embedded in saliva metabolites. Multiple logistic regression models yielded high area under the receiver-operating characteristic curves (AUCs) to discriminate healthy controls from each disease. The AUCs were 0.865 for oral cancer, 0.973 for breast cancer, 0.993 for pancreatic cancer, and 0.969 for periodontal diseases. The accuracy of the models was also high, with cross-validation AUCs of 0.810, 0.881, 0.994, and 0.954, respectively. Quantitative information for these 57 metabolites and their combinations enable us to predict disease susceptibility. These metabolites are promising biomarkers for medical screening

Effect of sample storage on stability of salivary glutathione, lipid peroxidation levels, and tissue factor activity

Saliva samples are often required to be stored for longer periods of time either because of the project protocol or because of lack of funding for analysis. The effects of 6 months storage (fresh, 30, 60, 90 120, 150, and 180d) on the stability of salivary reduced glutathione (GSH), lipid peroxidation (LPO) and 90days of storage (fresh, 15, 30, 60, and 90d) on the stability of salivary tissue factor (TF) activity and the stability of saliva imprint samples at 20 C were evaluated in this study. Salivary GSH, malondialdehyde (MDA) levels as an index of LPO, and TF activities were determined using the methods of Beutler, Yagi, and Quick, respectively. Saliva imprint samples were stained with Giemsa and microscopically examined. Salivary GSH levels and TF MDA levels increased significantly after 6 months of storage at -20 C. Leucocyte, epithelium and bacterium cell counts did not significantly change at the end of 90 d of storage. Saliva samples may be stored up to 1 month at -20 C for LPO assay. For cytological examinations, saliva samples may be stored for 90 d at -20 C. Further studies are needed to determine the stability of salivary GSH, and salivary TF activity stored less than 30 days at -20 C. On the other hand, if saliva samples are required to be stored, to avoid the changes because of different storage periods, we recommend that they must be stored under the same circumstances and in the same time period. J. Clin. Lab. Anal. 23:93-98, 2009. (C) 2009 Wiley-Liss, Inc

The relation between plasma tissue factor and oxidized LDL levels in acute coronary syndromes

Aim: Tissue factor (TF) is a low-molecular-weight glycoprotein responsible for the initiation of the coagulation cascade. The relation between oxidized low-density lipoprotein (Ox-LDL), that has been shown to be involved in atherogenesis, and TF has not been evaluated before in circulating plasma. The aim of this study was to determine plasma levels of TF and Ox-LDL in acute coronary syndrome (ACS) and stable coronary artery disease (SCAD). Methods: The study group consisted of 41 patients with ACS and 26 patients with SCAD. Among the ACS patients, 12 were diagnosed with unstable angina pectoris (UAP) and 29 were diagnosed with acute myocardial infarction (AMI). The control group consisted of 30 healthy volunteers. TF and Ox-LDL levels were evaluated by ELISA kits. Results: Ox-LDL levels were significantly higher in UAP and AMI patients compared with the control (p < 0.001) and SCAD (p < 0.01 and p < 0.001, respectively) groups. TF levels were significantly higher in the UAP, AMI and SCAD groups compared with the control group (p < 0.001, p < 0.001 and p < 0.01, respectively). In the AMI group a significant increase was observed in TF levels when compared with the SCAD group (p < 0.01). Plasma Ox-LDL levels were significantly and positively correlated with TF levels in the UAP and AMI groups (p < 0.05, r = 702, and p < 0.0001, r = 0.679, respectively). Conclusion: Thepotential link between Ox-LDL and TF in circulating blood in ACS may strengthen the evidence supporting a relationship between oxidant stress, lipids and thrombosis and consequently may contribute to understanding the mechanism through which Ox-LDL and TF may mediate the pathogenesis of CAD. Copyright © 2010 S. Karger AG, Basel