Fogle RL, Hollenbeak CS, Stanley BA, Vary TC, Kimball SR, Lynch CJ. Functional proteomic analysis reveals sexdependent differences in structural and energy-producing myocardial proteins in rat model of alcoholic cardiomyopathy. Physiol Genomics 43: 346 -356, 2011. First published January 18, 2011 doi:10.1152/physiolgenomics.00203.2010.-Long-term ethanol exposure leads to a sexually dimorphic response in both the susceptibility to cardiac pathology (protective effect of the female heart) and the expression of selected myocardial proteins. The purpose of the present study was to use proteomics to examine the effect of chronic alcohol consumption on a broader array of cardiac proteins and how these were affected between the sexes. Male and female rats were maintained for 18 wk on a 40% ethanol-containing diet in which alcohol was provided in drinking water and agar blocks. Differences in the content of specific cardiac proteins in isopycnic centrifugal fractions were determined using mass spectrometry on iTRAQlabeled tryptic fragments. A random effects model of meta-analysis was developed to combine the results from multiple iTRAQ experiments. Analysis of a network of proteins involved in cardiovascular system development and function showed that troponins were oppositely regulated by alcohol exposure in females (upregulated) vs. males (downregulated), and this effect was validated by Western blot analysis. Pathway analysis also revealed that alcohol-consuming males showed increased expression of proteins involved in various steps of oxidative phosphorylation including complexes I, III, IV, and V, whereas females showed no change or decreased content. One implication from these findings is that females may be protected from the toxic effects of alcohol due to their ability to maintain contractile function, maintain efficiency of force generation, and minimize oxidative stress. However, the alcohol-induced insult may lead to increased production of reactive oxygen species and structural abnormalities in male myocardium. iTRAQ; mass spectrometry; meta-analysis; random effects ALCOHOL ABUSE REMAINS ONE of the most common forms of drug abuse among both sexes in the United States. Although the risk of cardiovascular diseases is reduced with moderate alcohol consumption, potential consequences of excessive and chronic alcohol abuse, defined as Ͼ80 g of alcohol per day for longer than 5 yr, include heart dysfunction and heart failure (47). This quantity and frequency of ethanol consumption are sufficient to induce alcoholic heart muscle disease (AHMD) and, in approximately one-third of the cases, contributes to the development of a dilated cardiomyopathy (DCM: 48,50). AHMD is rarely the result of short-term ethanol intake but is more commonly observed in patients who consume excessive amounts of alcohol for prolonged periods (48, 50). This disease is clinically characterized by adverse alterations in both the structure and function of the myocardium, including ventricular dilation, thinning of the ventricular wall, and m...

Background Chronic alcohol abuse contributes not only to an increased risk of health-related complications, but also to a premature mortality in adults. Myocardial dysfunction, including the development of a syndrome referred to as alcoholic cardiomyopathy, appears to be a major contributing factor. One mechanism to account for the pathogenesis of alcoholic cardiomyopathy involves alterations in protein expression secondary to an inhibition of protein synthesis. However, the full extent to which myocardial proteins are affected by chronic alcohol consumption remains unresolved. Methods The purpose of this study was to examine the effect of chronic alcohol consumption on the expression of cardiac proteins. Male rats were maintained for 16 weeks on a 40% ethanol-containing diet in which alcohol was provided both in drinking water and agar blocks. Control animals were pair-fed to consume the same caloric intake. Heart homogenates from control- and ethanol-fed rats were labeled with the cleavable isotope coded affinity tags (ICAT™). Following the reaction with the ICAT™ reagent, we applied one-dimensional gel electrophoresis with in-gel trypsin digestion of proteins and subsequent MALDI-TOF-TOF mass spectrometric techniques for identification of peptides. Differences in the expression of cardiac proteins from control- and ethanol-fed rats were determined by mass spectrometry approaches. Results Initial proteomic analysis identified and quantified hundreds of cardiac proteins. Major decreases in the expression of specific myocardial proteins were observed. Proteins were grouped depending on their contribution to multiple activities of cardiac function and metabolism, including mitochondrial-, glycolytic-, myofibrillar-, membrane-associated, and plasma proteins. Another group contained identified proteins that could not be properly categorized under the aforementioned classification system. Conclusions Based on the changes in proteins, we speculate modulation of cardiac muscle protein expression represents a fundamental alteration induced by chronic alcohol consumption, consistent with changes in myocardial wall thickness measured under the same conditions.

Low selenium levels have been linked to a higher incidence of cancer and other diseases, including Keshan, Chagas, and Kashin-Beck, and insulin resistance. Additionally, muscle and cardiovascular disorders, immune dysfunction, cancer, neurological disorders, and endocrine function have been associated with mutations in genes encoding for selenoproteins. Selenium biology is complex, and a systems biology approach to study global metabolomics, genomics, and/or proteomics may provide important clues to examining selenium-responsive markers in circulation. In the current investigation, we applied a global proteomics approach on plasma samples collected from a previously conducted, double-blinded placebo controlled clinical study, where men were supplemented with selenized-yeast (Se-Yeast; 300 μg/day, 3.8 μmol/day) or placebo-yeast for 48 weeks. Proteomic analysis was performed by iTRAQ on 8 plasma samples from each arm at baseline and 48 weeks. A total of 161 plasma proteins were identified in both arms. Twenty-two proteins were significantly altered following Se-Yeast supplementation and thirteen proteins were significantly changed after placebo-yeast supplementation in healthy men. The differentially expressed proteins were involved in complement and coagulation pathways, immune functions, lipid metabolism, and insulin resistance. Reconstruction and analysis of protein-protein interaction network around selected proteins revealed several hub proteins. One of the interactions suggested by our analysis, PHLD-APOA4, which is involved in insulin resistance, was subsequently validated by Western blot analysis. Our systems approach illustrates a viable platform for investigating responsive proteomic profile in 'before and after' condition following Se-Yeast supplementation. The nature of proteins identified suggests that selenium may play an important role in complement and coagulation pathways, and insulin resistance.

Operation and effluent treatment costs are limiting factors for the success of recirculating aquaculture systems (RAS) in meeting seafood demand in the United States. Adopting a capture-and-reuse waste management model similar to terrestrial agriculture farmers would allow RAS farmers to monetize effluent and offset production costs. The moisture content and nutrient profile of RAS effluent makes it a potential option for use as a hydroponic fertilizer. Treatment of RAS waste is needed to mineralize particulate-bound nutrients before becoming a viable hydroponic nutrient solution. Anaerobic treatment (AT), a method used by municipal and agricultural waste treatment facilities to reduce total solids, has been shown to successfully mineralize particulate-bound nutrients from RAS effluent. Continuously mixed anaerobic batch bioreactors were used to evaluate the degree to which AT may mineralize particulate-bound nutrients in solid RAS waste. Concentrations of twelve different macro- and micro-nutrients were analyzed in the waste before and after treatment. Effluent samples were analyzed to determine the fraction of each nutrient in the solid and aqueous forms. This study showed that AT is an effective method to mineralize particulate-bound nutrients in RAS effluent and the mineralization rate data may be used to design a pilot-scaled flow-through RAS effluent treatment system.