1.
Nascent proteomes in peripheral blood mononuclear cells as a novel source for biomarker discovery in human stroke.
Bian, F, Simon, RP, Li, Y, David, L, Wainwright, J, Hall, CL, Frankel, M, Zhou, A
Stroke. 2014;(4):1177-9
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Abstract
BACKGROUND AND PURPOSE The proteome of newly synthesized proteins (nascent proteome) in peripheral blood mononuclear cells (PBMCs) can be a novel source of stroke biomarkers. Changes in the PBMC nascent proteome after stroke reflect the dynamic response-in-action not detectable in the total proteome (all existing proteins) in blood. Here, we test the application of nascent proteomics as a novel approach for stroke biomarker discovery. METHODS The PBMC nascent proteome in human blood was determined by metabolic labeling of fresh PBMC cultures with azidohomoalanine (an azide-containing methionine surrogate), followed by mass spectrometry detection and quantification of azidohomoalanine-labeled proteins. The PBMC nascent and total proteomes were compared between patients with stroke and matched controls. RESULTS Both PBMC nascent and total proteomes showed differences between stroke patients and controls. Results of hierarchical clustering analysis of proteomic data revealed greater changes in the nascent than in the total PBMC proteomes, supporting the usefulness of the PBMC nascent proteome as a novel source of stroke biomarkers. CONCLUSIONS Nascent proteomes in PBMC can be a novel source for biomarker discovery in human stroke.
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Potential mechanisms of the acute coronary syndrome presentation in patients with the coronary slow flow phenomenon - insight from a plasma proteomic approach.
Kopetz, VA, Penno, MA, Hoffmann, P, Wilson, DP, Beltrame, JF
International journal of cardiology. 2012;(1):84-91
Abstract
AIMS: The coronary slow flow phenomenon [CSFP] is a coronary microvascular disorder, characterized by delayed distal vessel opacification despite the absence of obstructive coronary artery disease. Patients frequently present with an acute coronary syndrome [ACS] although the pathophysiological mechanisms responsible are unknown. The aim of this study was to identify potential mechanisms for the ACS presentation associated with the CSFP using a plasma proteomic profiling approach. METHODS AND RESULTS Plasma samples from nine CSFP subjects [56 ± 11years] were assayed for high sensitivity C-reactive protein [hsCRP], troponin T [TnT], creatine kinase [CK], and proteomic analyses (n=6), during an ACS presentation and one month later [chronic phase]. Proteomic analysis involved chromatographic depletion of abundant plasma proteins followed by two-dimensional differential gel electrophoresis [2-D DIGE]. Protein spots demonstrating ±1.5-fold change relative to the control were identified by mass spectrometry and two differentially expressed proteins were selected for validation via Western blotting. During the ACS presentation, hsCRP was elevated [ACS=14.9 ± 3.9 mg/L vs chronic=4.23 ± 1.37 mg/L, p=0.05] but TnT and CK levels were unchanged. Proteomic analysis identified six proteins that were significantly different in abundance between the acute and chronic samples. During the ACS presentation there was a 1.6 ± 0.13 fold increase in the anti-oxidant enzyme paraoxonase-1 and an increase in inflammatory proteins alpha-1-antichymotrypsin [1.65 ± 0.13 fold] and alpha-1-antitrypsin [2.5 ± 0.34 fold]. The latter was confirmed by Western blotting [1.33 ± 0.17 OD acute/chronic ratio, p=0.05]. CONCLUSION The findings from this novel detailed approach, implicate an inflammatory/oxidative stress process in the pathogenesis of the ACS presentation associated with the CSFP. Future studies should further elucidate these mechanisms.
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Effect of high dose thiamine on the levels of urinary protein biomarkers in diabetes mellitus type 2.
Riaz, S, Skinner, V, Srai, SK
Journal of pharmaceutical and biomedical analysis. 2011;(4):817-25
Abstract
The proteomics is known to be a valuable field of study and has become one of the most attractive sub-disciplines in clinical proteomics for human diseases. In the present research work, the levels of urinary protein biomarkers of diabetes mellitus type 2 using proteomic technology have been identified and characterized. Effect of high dose thiamine has also been observed on the levels of these marker proteins. Above 100 type 2 diabetic patients, and 50 same age and sex-matched normal healthy controls were recruited from the Sheikh Zayed Hospital, Lahore, Pakistan and 40 diabetic and 20 control have completed the trial. The urine samples from control and diabetic groups before or after thiamine therapy were further analyzed and identified by 2-D liquid chromatographic system (HPLC) and mass spectrometry MALDI-TOF/TOF and microTOF analysis. All the samples belonging to the control and diabetic groups were then analyzed by ELISA and estimated the levels of some proteins which were found to vary. In the urine samples, the levels of transthyretin, AMBP, haptoglobin precursor were found to decrease while albumin, zinc α 2 glycoprotein, RBP4 and E cadherin were found to increase in the diabetic patients as compared to the controls. The level of albumin in the urine samples of diabetic patients only decreased by 34% after thiamine therapy as compared to the controls and the placebo, while other urinary protein markers did not show a significant change after the therapy. Assessment of the levels of these biomarkers will be helpful in the diagnosis and treatment of diabetes mellitus type 2.