Differential Protein Expression in Berry Skin from Red Grapes with Varying Hybrid Character

Protein expression from the berry skin of four red grape biotypes with varying hybrid character was compared at a proteome-wide level to identify the metabolic pathways underlying divergent patterns of secondary metabolites. A bottom-up shotgun proteomics approach with label-free quantification and MaxQuant-assisted computational analysis was applied. Red grapes were from (i) purebred Vitis vinifera (Aglianico cv.); (ii) V. vinifera (local Sciascinoso cv.) grafted onto an American rootstock; (iii) interspecific hybrid (V. vinifera × V. labrusca, Isabel), and (iv) uncharacterized grape genotype with hybrid lineage, producing relatively abundant anthocyanidin 3,5-O-diglucosides. Proteomics supported the differences between hybrids and purebred V. vinifera grapes, consistently with distinct phenotypic metabolite assets. Methanol O-anthraniloyltransferase, which catalyses the synthesis of methyl anthranilate, primarily responsible for the "foxy" odour, was exclusive of the Isabel hybrid grape. Most of the proteins with different expression profiles converged into coordinated biosynthetic networks of primary metabolism, while many possible enzymes of secondary metabolism pathways, including 5-glucosyltransferases expected for hybrid grapes, remained unassigned due to incomplete protein annotation for the Vitis genus. Minor differences of protein expression distinguished V. vinifera scion grafted onto American rootstocks from purebred V. vinifera skin grapes, supporting a slight influence of the rootstock on the grape metabolism

Sensory profile of P.D.O. Mozzarella di Bufala Campana Cheese

The aim of this work was to define the typical sensory profile of DOP Mozzarella di Bufala Campana cheese as related to the technological process (industrial or artisanal), the geographical area (Salerno or Caserta), the season (spring or summer) and to the chemical composition of both milk and cheese. The results obtained showed that the sensorial profile of Mozzarella DOP changed for each variable considered. In particular, Caserta samples were more salty, with a more intense butter odour, higher peelability and homogeneity surface than those from Salerno area; the industrial samples, compared to the artisanal ones, were more salty, hard and "chewable", and exhibited an external layer more difficult to remove and more homogeneous

Excretion of dietary cow’s milk derived peptides into breast milk

Nanoflow-HPLC-tandem mass spectrometry (MS/MS) was used to analyze the peptide fraction of breast milk samples collected from a single non-atopic donor on different days (ten samples) after receiving an oral load of cow’s milk (by drinking 200 mL of bovine milk). In addition, breast milk was sampled from the same lactating mother over a 6-h period at 5 time points after drinking cow’s milk. We aimed to trace the intra-individual variability and to define a time profile of the excretion of dietary peptides into breast milk. Overall, 21 peptides exclusively originating from both bovine caseins and whey proteins with no match within the human milk proteome were identified in the breast milk samples. These peptides were missing in the breast milk obtained from the mother after a prolonged milk- and dairy-free diet (three samples). The time course of cow’s milk-derived β-Lg f(125-135) and β-casein f(81-92) in breast milk

Proteomic study of muscle sarcoplasmic proteins using AUT-PAGE/SDS-PAGE as two-dimensional gel electrophoresis

The conversion of muscle to meat in pig involves mainly proteolysis of myofibrillar proteins, which undergo notable changes since early stage of rigor mortis, even after 48 h post mortem. The tenderness of meat has been thoroughly investigated to understand the biochemical mechanisms, which influence texture and flavour development as well as the technological parameters and hence meat quality. Cytoplasmic proteolytic calcium dependent enzymes, named -and m-calpains, which act in the early stages of rigor mortis, significantly contribute to tenderization weakening myofibrils. These enzymes, however, act for fewdays because they are specifically inhibited by calpastatin and by pH lowering. However, when pH falls to about 5.0, proteolytic activity on muscle proteins is continued by longer acting lysosomal proteinase, cathepsins [3,7–9]. Post mortem proteolysis also causes relevant changes in sarcoplamic protein fraction, which represent the water soluble fraction (quantitatively about 30–35%) of meat total protein, and the involved proteins has already been identified by proteomic-based studies. Recent investigations have demonstrated that the most commonly found Lactobacillus species in dry fermented meats are able to hydrolyse myofibrillar and sarcoplasmic muscle proteins in vitro.The most abundant sarcoplasmic proteins, as mixture of basic polypeptides with a narrow spread range of molecular masses, represented an excellent model to test our analytical technique and to delineate its capabilities. In the present study, we compared 2D AUT-PAGE/SDSPAGE maps of water-soluble proteins extracted from fresh meat and from dry-cured ham, a non fermented product, from “Naples-type” salami, a microbiologically fermented product, and from “Coppa”, a typical semi-fermented product. Electrophoretically separated proteins have been identified by MALDI-ToF mass fingerprinting

Caching Placement Strategies for Dynamic Content Delivery in Metro Area Networks

Video-on-Demand (VoD) traffic explosion has been one of the main driving forces behind the recent Internet evolution from a traditional connection-centric architecture towards the new content-centric paradigm. To cope with this evolution, caching of VoD contents closer to the users in core, metro and even metro-access optical network equipment is regarded to be a prime solution that could help mitigating this traffic growth. However, the optimal caches placement and dimensioning is not univocal, especially in the context of a dynamic network, as it depends on various parameters, such as network topology, users behavior and content popularity. In this paper, we focus on a dynamic VoD content delivery scenario in a metropolitan network implementing different caching strategies. We evaluate the performance of the various caching strategies in terms of network-capacity occupation showing the savings in resource occupation in each of the network segments. We also evaluate the effect of the distribution of the storage capacity on the overall average number of hops of all requests. The obtained numerical results show that, in general, a significant amount of network resources can be saved by enabling content caching near to end-users. Moreover, we show that blindly providing caching capability in access nodes may result unnecessary, whereas a balanced storage distribution between access and metro network segments provides the best performance

Replacing Maize Grain with Ancient Wheat Lines By-Products in Organic Laying Hens' Diet Affects Intestinal Morphology and Enzymatic Activity

The effects of replacement of maize grain with ancient wheat by-products on intestinal morphometry and enzymatic activity in laying hens was studied. Eighty hens were divided into two groups (40 each, 8 replicates, 5 hens/replicate) fed two isoproteic and isoenergetic diets. In the treated group, part of the maize was replaced by a mix of ancient grains (AGs) middling, in a 50:50 ratio of Triticum aestivum L. var. spelta (spelt) and Triticum durum dicoccum L. (emmer wheat). The AG diet affected the weight of all the large intestine tracts, decreasing the weight of caeca (p < 0.01) and increasing those of colon (p < 0.01), rectum and cloaca (p < 0.05). Villus height in the AG group was higher (p < 0.01) than the control for the duodenum and jejunum, while for the ileum, the control group showed the highest values (p < 0.01). The submucosa thickness was higher (p < 0.01) in the control group for the duodenum and ileum, while the jejunum for the AG group showed the highest (p < 0.05) submucosa thickness. The crypts depth was higher (p < 0.01) in the control group for the duodenum and ileum. Enzyme activity was enhanced by AGs (p < 0.01) in the duodenum. Regarding the jejunum, sucrase-isomaltase and alkaline phosphatase had higher activity (p < 0.05 and p < 0.01, respectively) in the AG group. In the ileum, sucrase-isomaltase showed higher activity (p < 0.01) in the control group, while alkaline phosphatase showed the highest values (p < 0.05) in the AG group. Overall, results suggested that the dietary inclusion of AGs exerted positive effects in hens, showing an improved intestinal function

Replacing Maize Grain with Ancient Wheat Lines By-Products in Organic Laying Hens’ Diet Affects Intestinal Morphology and Enzymatic Activity

The effects of replacement of maize grain with ancient wheat by-products on intestinal morphometry and enzymatic activity in laying hens was studied. Eighty hens were divided into two groups (40 each, 8 replicates, 5 hens/replicate) fed two isoproteic and isoenergetic diets. In the treated group, part of the maize was replaced by a mix of ancient grains (AGs) middling, in a 50:50 ratio of Triticum aestivum L. var. spelta (spelt) and Triticum durum dicoccum L. (emmer wheat). The AG diet affected the weight of all the large intestine tracts, decreasing the weight of caeca (p < 0.01) and increasing those of colon (p < 0.01), rectum and cloaca (p < 0.05). Villus height in the AG group was higher (p < 0.01) than the control for the duodenum and jejunum, while for the ileum, the control group showed the highest values (p < 0.01). The submucosa thickness was higher (p < 0.01) in the control group for the duodenum and ileum, while the jejunum for the AG group showed the highest (p < 0.05) submucosa thickness. The crypts depth was higher (p < 0.01) in the control group for the duodenum and ileum. Enzyme activity was enhanced by AGs (p < 0.01) in the duodenum. Regarding the jejunum, sucrase-isomaltase and alkaline phosphatase had higher activity (p < 0.05 and p < 0.01, respectively) in the AG group. In the ileum, sucrase-isomaltase showed higher activity (p < 0.01) in the control group, while alkaline phosphatase showed the highest values (p < 0.05) in the AG group. Overall, results suggested that the dietary inclusion of AGs exerted positive effects in hens, showing an improved intestinal function

Antibody-independent identification of bovine milk-derived peptides in breast-milk

Exclusively breast-fed infants can exhibit clear signs of IgE or non IgE-mediated cow's milk allergy. However, the definite characterization of dietary cow's milk proteins (CMP) that survive the maternal digestive tract to be absorbed into the bloodstream and secreted into breast milk remains missing. Herein, we aimed at assessing possible CMP-derived peptides in breast milk. Using high performance liquid chromatography (HPLC)-high resolution mass spectrometry (MS), we compared the peptide fraction of breast milk from 12 donors, among which 6 drank a cup of milk daily and 6 were on strict diary-free diet. We identified two bovine β-lactoglobulin (β-Lg, 2 out 6 samples) and one α(s1)-casein (1 out 6 samples) fragments in breast milk from mothers receiving a cup of bovine milk daily. These CMP-derived fragments, namely β-Lg (f42-54), (f42-57) and α(s1)-casein (f180-197), were absent in milk from mothers on dairy-free diet. In contrast, neither intact nor hydrolyzed β-Lg was detected by Western blot and competitive ELISA in any breast milk sample. Eight additional bovine milk-derived peptides identified by software-assisted MS were most likely false positive. The results of this study demonstrate that CMP-derived peptides rather than intact CMP may sensitize or elicit allergic responses in the neonate through mother's milk. Immunologically active peptides from the maternal diet could be involved in priming the newborn's immune system, driving tolerogenic response