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Variation in water-holding capacity (WHC), which presents a major economic burden to the swine industry, is considered to be underpinned by variation at a molecular and biochemical level. High-resolution 2D DIGE followed by MS analysis and Western blot were used to unravel the proteome of muscle exudate, collected following centrifugation, in the pH 4-7 range. A first 2DE-based protein map of this substrate was produced where 89 spots were successfully characterised. Two phenotypes divergent for WHC plus one intermediate were compared with a view to deciphering the biochemical processes impacting on variation in WHC. Twenty spots were observed to be altered across the phenotypes. Of these, 14 represented sixteen proteins including metabolic enzymes, stress response proteins and structural proteins. Triosephosphate isomerase and transferrin showed a major difference between the two extreme phenotypes, and may have potential as biological markers for WHC prediction. Several members of the HSPs family were highlighted. This proteomic study makes an important contribution towards a more detailed molecular view of the processes behind WHC and will provide a valuable resource for future investigations.

Multidrug resistance (MDR) is a serious obstacle to efficient cancer treatment. Overexpression of P-glycoprotein (P-gp) plays a significant role in MDR. Recent studies proved that targeting cellular metabolism could sensitize MDR cells. In addition, metabolic alterations could affect the extracellular vesicles (EVs) cargo and release. This study aimed to: i) identify metabolic alterations in P-gp overexpressing cells that could be involved in the development of MDR and, ii) identify a potential role for the EVs in the acquisition of the MDR. Two different pairs of MDR and their drug-sensitive counterpart cancer cell lines were used. Our results showed that MDR (P-gp overexpressing) cells have a different metabolic profile from their drug-sensitive counterparts, demonstrating decreases in the pentose phosphate pathway and oxidative phosphorylation rate; increases in glutathione metabolism and glycolysis; and alterations in the methionine/S-adenosylmethionine pathway. Remarkably, EVs from MDR cells were capable of stimulating a metabolic switch in the drug-sensitive cancer cells, towards a MDR phenotype. In conclusion, obtained results contribute to the growing knowledge about metabolic alterations in MDR cells and the role of EVs in the intercellular transfer of MDR. The specific metabolic alterations identified in this study may be further developed as targets for overcoming MDR.

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Grape pomace (GP) represents the main solid by-product deriving from grape processing. The aim of this study was to evaluate the effect of dietary GP intake on nutritional quality, lipid oxidation and volatile profile of chicken meat. A total of 112 Ross 508 broilers were randomly divided into 4 groups and fed for 21 days with a standard diet. For the remaining 28 days of the trial, the control group (CG) continued to receive a standard diet, while the experimental groups (EGs) were fed with diets containing different GP concentrations: 2.5% (EG1), 5% (EG2) and 7% (EG3). Following the slaughtering, samples of breast meat were collected from each group. No significant differences were observed for pH, cooking loss and meat brightness, whereas the GP intake showed effectiveness in inducing variations in drip loss, meat yellowness and redness. The experimental feeding strategy also induced changes in the fatty acid profile, with an overall increase in polyunsaturated fatty acids (PUFA), mainly due to the increase in concentration of linoleic acid. The dietary supplementation also induced a decrease in lipid oxidation in meat, a finding also confirmed by the reduction in volatile aldehydes in 7 days stored raw meat. The feeding strategy based on the use of GP did not induce detrimental effects on the quality of broiler meat and showed the potential to lengthen the shelf-life as a direct consequence of the improvement in the oxidative stability. Overall, the present study showed a viable way for the recovery and the valorization of an agro-industrial by-product, with potential benefits also from an environmental point of view.

BackgroundMeat quality is a complex trait influenced by a range of factors with post mortem biochemical processes highly influential in defining ultimate quality. High resolution two-dimensional DIfference Gel Electrophoresis (2-D DIGE) and Western blot were applied to study the influence of post mortem meat ageing on the proteome of pork muscle. Exudate collected from the muscle following centrifugation was analysed at three timepoints representing a seven day meat ageing period.ResultsThe intensity of 136 spots varied significantly (p < 0.05) across this post mortem period and 40 spots were identified using mass spectrometry. The main functional categories represented were metabolic proteins, stress-related proteins, transport and structural proteins. Metabolic and structural proteins were generally observed to increase in abundance post mortem and many likely represent the accumulation of the degradation products of proteolytic enzyme activity. In contrast, stress-related proteins broadly decreased in abundance across the ageing period. Stress response proteins have protective roles in maintaining cellular integrity and a decline in their abundance over time may correlate with a reduction in cellular integrity and the onset of meat ageing. Since cellular conditions alter with muscle ageing, changes in solubility may also contribute to observed abundance profiles.ConclusionsMuscle exudate provided valuable information about the pathways and processes underlying the post mortem ageing period, highlighting the importance of post mortem modification of proteins and their interaction for the development of meat quality traits.

The aim of this study was to evaluate the effect of dietary supplementation with dried grape pomace on beef quality. Ten Friesian calves were divided into two groups, a control group that received a standard diet, and an experimental group that was administered the dietary supplementation. At the end of the 75 days of the trial, animals were slaughtered, and meat samples analyzed for physical and chemical properties, fatty acids composition, lipid oxidation, volatile compounds, and biogenic amines. The fatty acid profile resulted affected by dietary supplementation, since an increase in concentration of linoleic acid was observed. Furthermore, a reduction of lipid oxidation was found in the same samples. With reference to volatile compounds a reduction of hexanal and an increase of 2‐3 octanedione was evidenced, while no effects were induced by diets on the synthesis of biogenic amines. The grape pomace exploitation as a dietary supplement in bovine diet did not have negative effects on the quality of beef and showed the potential to extend shelf life due to marked improvement in oxidative stability. Overall, the present study showed a viable way for the recovery and the valorization of the main by-product of the oenological industry.

Two dimensional Difference Gel Electrophoresis (2-D DIGE) and mass spectrometry were applied to investigate the changes in metabolic proteins that occur over a seven day (day 1, 3 and 7) post mortem ageing period in porcine centrifugal exudate from divergent meat quality phenotypes. The objectives of the research were to enhance our understanding of the phenotype (water holding capacity) and search for biomarkers of this economically significant pork quality attribute. Major changes in protein abundance across nine phenotype-by-time conditions were observed. Proteomic patterns were dominated by post mortem ageing timepoint. Using a machine learning algorithm (l1-regularized logistic regression), a model was derived with the ability to discriminate between high drip and low drip phenotypes using a subset of 25 proteins with an accuracy of 63%. Models discriminating between divergent phenotypes with accuracy of 72% and 73% were also derived comparing respectively, high drip plus intermediate phenotype (considered as one phenotype) versus low drip and comparing low drip plus intermediate phenotype (considered as one phenotype) versus high drip. In all comparisons, the general classes of discriminatory proteins identified include metabolic enzymes, stress response, transport and structural proteins. In this research we have enhanced our understanding of the protein related processes underpinning this phenotype and provided strong data to work toward development of protein biomarkers for water holding capacity.