The aim of this work was to characterize UV-absorbing compounds (UAC) in milk in relation to diet. In winter, 4 groups of cows each received a different diet: concentrate rich containing 35% cocksfoot hay (CCH), maize silage (MS), rye grass silage (RS), or rye grass hay (RH). In summer, 2 additional diets were given: mountain grassland hay (GH) and mountain grassland pasture (GP). Polyphenols were analyzed by HPLC and Folin reaction on forages and UAC were extracted from milks and analyzed by HPLC. In forages, the number of polyphenols was lowest in MS (57) and greatest in GP (85). Twenty-four peaks were identified, accounting for 28 to 47% peak area at 280 nm. Caffeoyl compounds and flavonoid glycosides were mainly found in RH, GH, and GP. Hydrolyzed compounds such as hydroxycinnamic acids and aglycones were found in MS and RS. Estimated amounts of polyphenols were lowest for MS (3.7 g/kg), roughly similar for CH, RS, and RH (about 15 g/kg), and greatest for GH and GP (21.6 and 35.3 g/kg, respectively). About 230 different peaks were separated in milks. Milks from RH and GP contained the lowest (87) and the greatest (127) numbers of peaks, respectively. Only 10 peaks were identified, accounting for 21 to 54% of the total spectra area. In addition to the major compound hippuric acid, phenylacetic acid, benzoic acid, 4-hydroxybenzoic acid benzaldehyde, catechol, and small amounts of ferulic acid were found in varying amounts depending on the diet. Flavonoids such as quercetin, luteolin, and apigenin were also present. Hippuric acid was clearly related to the presence of caffeoylquinic compounds in forages. Other identified UAC may originate essentially from forage simple polyphenols or from cell wall aromatics. Some of the several unknown compounds may also originate from the transformation of other nutrients. Estimated amounts of UAC were widely variable within each animal group. They were surprisingly high in CCH and roughly similar in all milks from preserved forages (about 3.6 mg/L), with generally greater values for GH milks, whereas the greatest amount was found in GP milks (13.3 mg/L). Hierarchical clustering clearly discriminated the 6 diets, showing that there were major differences in GP milks. Some UAC were specific to one or a group of diets. Ultraviolet-absorbing compounds are therefore a potential tool to distinguish between milks according to diet. In addition, they may have a bioactive effect on milk component conservation or on human health.
Phenolics contribute to the micronutrient composition of forages, which in turn may affect animal product composition. To assess the importance of these compounds, the polyphenolic and botanical composition of a permanent mountain pasture in the Massif Central (France) were studied at three stages of growth. Phenolic fractions (acids, flavonoids and total) were analysed using HPLC and specific colorimetric methods over the whole pasture and in nine main species. On a botanical level, 43 species including 31 dicotyledons composed a mountain variant of Cynosurion grouping. Over time, the species growing on the field changed, the main plants being Festuca nigrescens and eight dicotyledons. In whole pasture, about 170 different compounds were separated by HPLC, of which only 30 were common to all stages. The total polyphenolic content was estimated as 31, 32 and 19 g kg −1 DM for the three stages. They were mainly composed of dihydroxycinnamic derivatives (3,5-di-O-caffeoylquinic, chlorogenic, 1,5-di-O-caffeoylquinic acids), and of flavonoids in lesser proportions. Some polyphenols were peculiar to a few species and others were ubiquitous. The two major dicaffeoylquinic acids were identified for the first time in Achillea millefolium and Knautia arvernensis. Colorimetric measurements of phenolic fractions were in good agreement with HPLC analysis (P < 0.01) for all species studied. Thus, on the mountain pasture studied, a cow would be expected to ingest up to 500 g of polyphenols daily, which could affect the quality of animal products.
Fat loss during cooking of duck "foie gras" is the main quality issue for both processors and consumers. Despite the efforts of the processing industry to control fat loss, the variability of fatty liver cooking yield remains high and uncontrolled. To better understand the biological basis of this phenomenon, a proteomic study was conducted. To analyze the protein fraction soluble at low ionic strength (LIS), we used bidimensional electrophoresis and mass spectrometry for the identification of spots of interest. To analyze the protein fraction not soluble at low ionic strength (NS), we used the shotgun strategy. The analysis of data acquired from both protein fractions suggested that at the time of slaughter, livers with low fat loss during cooking were still in anabolic processes with regard to energy metabolism and protein synthesis, whereas livers with high fat loss during cooking developed cell protection mechanisms. The variability in the technological yield observed in processing plants could be explained by a different physiological stage of liver steatosis.
Sarcopenia corresponds to the loss of muscle mass occurring during aging, and is associated with a loss of muscle functionality. Proteomic links the muscle functional changes with protein expression pattern. To better understand the mechanisms involved in muscle aging, we performed a proteomic analysis of Vastus lateralis muscle in mature and older women. For this, a shotgun proteomic method was applied to identify soluble proteins in muscle, using a combination of high performance liquid chromatography and mass spectrometry. A label-free protein profiling was then conducted to quantify proteins and compare profiles from mature and older women. This analysis showed that 35 of the 366 identified proteins were linked to aging in muscle. Most of the proteins were under-represented in older compared with mature women. We built a functional interaction network linking the proteins differentially expressed between mature and older women. The results revealed that the main differences between mature and older women were defined by proteins involved in energy metabolism and proteins from the myofilament and cytoskeleton. This is the first time that label-free quantitative proteomics has been applied to study of aging mechanisms in human skeletal muscle. This approach highlights new elements for elucidating the alterations observed during aging and may lead to novel sarcopenia biomarkers. Molecular & Cellular Proteomics 13:
Listeria monocytogenes is a foodborne pathogen that can cause invasive severe human illness (listeriosis) in susceptible patients. Most human listeriosis cases appear to be caused by consumption of refrigerated ready-to-eat foods. Although initial contamination levels in foods are usually low, the ability of these bacteria to survive and multiply at low temperatures allows it to reach levels high enough to cause disease. This study explores the set of proteins that might have an association with L. monocytogenes adaptation to different temperatures. Cultures were grown in biofilm, the most widespread mode of growth in natural and industrial realms. Protein extractions were performed from three different growth temperatures (10, 25, and 37°C) and two growth phases (early stage and mature biofilm). L. monocytogenes subproteomes were targeted using three extraction methods: trypsin-enzymatic shaving, biotin-labeling and cell fractionation. The different subproteomes obtained were separated and analyzed by shotgun proteomics using high-performance liquid chromatography combined with tandem mass spectrometry (LC-OrbiTrap LTQVelos, ThermoFisher Scientific). A total of 141 (biotinylation), 98 (shaving) and 910 (fractionation) proteins were identified. Throughout the 920 unique proteins identified, many are connected to basic cell functions, but some are linked with thermoregulation. We observed some noteworthy protein abundance shifts associated with the major adaptation to cold mechanisms present in L. monocytogenes , namely: the role of ribosomes and the stressosome with a higher abundance of the general stress protein Ctc (Rl25) and the general stress transcription factor sigma B (σ B ), changes in cell fluidity and motility seen by higher levels of foldase protein PrsA2 and flagellin (FlaA), the uptake of osmolytes with a higher abundance of glycine betaine (GbuB) and carnitine transporters (OpucA), and the relevance of the overexpression of chaperone proteins such as cold shock proteins (CspLA and Dps). As for 37°C, we observed a significantly higher percentage of proteins associated with transcriptional or translational activity present in higher abundance upon comparison with the colder settings. These contrasts of protein expression throughout several conditions will enrich databases and help to model the regulatory circuitry that drives adaptation of L. monocytogenes to environments.
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