Quality Evaluation of Fresh Pasta Fortified with Sourdough Containing Wheat Germ and Wholemeal Semolina.

Pasta is a staple food in the Mediterranean diet, primarily manufactured with two essential ingredients, semolina and water; nowadays, it is often supplemented with functional ingredients. In this work, a sourdough obtained with wheat germ and wholemeal semolina was used, in order to improve sensorial and nutritional properties of fresh pasta, to prevent lipids oxidation, and to improve the shelf life. Three different formulations were prepared, a first one using semolina, a second one with raw wheat germ, wholemeal semolina, and semolina, and the last one with semolina and sourdough. The study highlighted the improved nutritional properties of pasta with sourdough (reduced phytic acid content, higher antioxidant activity and phenolic content). Proteins, ashes, dietary fibers, lipids, and tocols (vitamin E) increased in pasta with wheat germ and wholemeal semolina, and with sourdough. The amount of tocols decreased in pasta samples after cooking, except for the ß-tocopherol in sourdough pasta, the amount of which remained high, surprisingly. Lipase and lipoxygenase enzymes likely decreased as an effect of the pasteurization process. The NMR analysis showed that lipid oxidation was higher in semolina pasta than in pasta with wheat germ, most likely due to the protective effect of antioxidants deriving from wheat germ.

In Vivo and In Vitro Starch Digestibility of Fresh Pasta Produced Using Semolina-Based or Wholemeal Semolina-Based Liquid Sourdough.

The use of wholemeal flour and sourdough fermentation in different food matrices has received considerable attention in recent years due to its resulting health benefits. In this study, a semolina-based and a wholemeal semolina-based sourdough were prepared and added to the formulation of gnocchetti-type fresh pasta. Four types of gnocchetti were made, using semolina plus semolina-based sourdough (SS), semolina plus wholemeal semolina-based sourdough (SWS), semolina alone (S), and semolina plus wholemeal semolina (WS). The latter two were used as controls. The digestibility of starch was studied both in vitro and in vivo, and the glycemic response (GR) and glycemic load (GL) were determined. Starch digestibility, both in vivo and in vitro, was higher in wholemeal semolina than semolina pasta and the resulting GR values (mg dL-1 min-1) were also higher (2209 and 2277 for WS and SWS; 1584 and 1553 for S and SS, respectively). The use of sourdough significantly reduced the rapidly digestible starch (RDS) content and increased the inaccessible digestible starch (IDS) content. The addition of sourdough to the formulation had no effect on the GR values, but led to a reduction of the GL of the pasta. These are the first data on the GR and GL of fresh pasta made with sourdough.

Impact of Almond Variety on "Amaretti" Cookies as Assessed through Image Features Modeling, Physical Chemical Measures and Sensory Analyses.

In the present study, the influence of almond variety on color, chemical, physical and sensory characteristics of "amaretti" cookies during the shelf life, was assessed. Four varieties were chosen for the study, two of which were local (Cossu, Arrubia) and two widely cultivated (Tuono, Texas). Almonds have been characterized in the content of proteins, crude fat, amygdalin and fatty acids profile. The evolution of the characteristics during the shelf life hasbeen measured through image data modeling, texture, physical chemical and sensory analyses. Data were then treated with a multivariate approach performing a PCA. Image analysis and fitting on log normal and powerlaw functions highlighted the influence of the variety on the total area affected by surface breakages, and on the distribution of the cracking surfaces dimension classes. Texture parameters (crust hardness, thickness and work of deformation) were negatively correlated to moisture content. Sensory profile confirmed the differences in tactile features measured through instrumental texture, while slight to no differences were found in odor profile. Consumer test showed an higher acceptability for Arrubia, Texas and Tuono samples throughout the shelf life, while Cossu samples were less accepted. Overall, the choice of almond variety influences product features and liking of almond products, therefore it represents an important phase to direct the choice of both farmers and confectionery manufacturers.

Fresh Pasta Manufactured with Fermented Whole Wheat Semolina: Physicochemical, Sensorial, and Nutritional Properties.

Fresh pasta (SP) was prepared by mixing semolina with liquid sourdough, whole wheat semolina based, and the effects of sourdough inclusion were evaluated against a control sample (CP) prepared using semolina and whole wheat semolina. Physicochemical, nutritional, and sensorial analyses were performed on pasteurized fresh pasta, before and after cooking. The optimum cooking time was not affected by whole wheat sourdough, whereas differences were found in color, firmness, and cooking loss. Changes of in vitro digested starch fractions in SP pasta were affected by a higher cooking loss. Overall, SP samples were characterized by improved nutraceutical features, namely higher content of free essential amino acids and phenolic compounds, lower phytic acid content, and higher antioxidant activity. Sensory analyses (acceptability and check-all-that-apply (CATA) tests) showed significantly higher scores for the SP, and the differences were enhanced when the consumers were informed about the product composition and how it was manufactured. Consumers checked for more positive sensory parameters for the SP than the CP.

Effect of liquid sourdough technology on the pre-biotic, texture, and sensory properties of a crispy flatbread.

The present study compared liquid sourdough technology with baker's yeast leavening when applied to the production of a semolina-based crispy flatbread. Following in vitro starch digestion, the results revealed the sourdough leavened flatbread to contain a lower percentage of rapidly digestible starch (16%), higher amounts of slowly digestible starch (27%) and inaccessible digestible starch (4.1%) compared with the baker's yeast leavened flatbread (20, 20, and 2.4%, respectively), making the former nutritionally healthier. The sourdough leavened bread was crispier, stiffer and more solid, as shown by texture analyses, although Raman spectroscopy revealed no differences in the crystallinity status of starch. The descriptive analyses show that the use of sourdough enhances the positive sensory traits, as rated by the consumer panel scores (6.08 vs. 5.56). In summary, the results indicate that the implementation of sourdough technology in the production of flat crispy breads could confer economic advantages to this product.

The role of fatty acids and triglycerides in the gonads of Paracentrotus lividus from Sardinia: Growth, reproduction and cold acclimatization.

A detailed characterization of lipid extracts from gonads of P. lividus over a year has been performed combining GC and NMR measurements. For this purpose, sea urchins from two different Sardinian coastal areas were collected monthly. The results underlined a correlation between gonad fatty acids profiles and both water temperature and reproduction process. In particular, EPA and ARA appear to be the most altered fatty acids following seasonal changes. It is suggested that EPA could represent a biomarker of reproduction, reaching a content around 14% during gametogenesis, mainly due to an increase of the sn-1,3 position in TAGs. On the contrary, ARA seems to play a more important role in response to cold acclimatization, which is reflected in a modulation of ARA content in the sn-1,3 position in TAGs.

Molecular details on gilthead sea bream (Sparus aurata) sensitivity to low water temperatures from 1H NMR metabolomics.

Biometric and metabolic responses of gilthead sea bream to cold challenge are described following a growth trial divided into three water temperature steps, namely cooling, cold maintenance and recovery. Experimental data provide a useful description of fish response to thermal stress at both zootechnical and molecular level. Although no mortality has been observed, Nuclear Magnetic Resonance-based metabolomics confirms the marked sensitivity of this fish species to low water temperature, and explains some key molecular events associated to fish response to cold. Increase in hepatosomatic index is associated to liver fat accumulation, as a consequence of lipid mobilization from muscle and other extrahepatic tissues, and metabolic rearrangements linked to homeoviscous adaptation of cellular membranes are observed. Following primary responses to descending temperature from 18°C to 11°C, the energetic metabolism (insulin signaling, glycolysis) is first clearly affected; then, at constant low water temperature (11°C), the most perturbed metabolic pathways are related to methionine cycle in liver, while osmoregulatory function is exerted by TMAO in muscle. Water temperature recovery from 11°C to 18°C stimulates gluconeogenesis and glycogen synthesis activities at hepatic level, although the rate of a thermo-compensatory response seems to be slower than that of the cooling phase. The obtained results are intended to guide novel high-performance feed formulations for gilthead sea bream reared during winter.

Proteomic dataset of Paracentrotus lividus gonads of different sexes and at different maturation stages.

We report the proteomic dataset of gonads from wild Paracentrotus lividus related to the research article entitled "Proteomic changes occurring along gonad maturation in the edible sea urchin Paracentrotus lividus" [1]. Gonads of three individuals per sex in the recovery, pre-mature, mature, and spent stages were analyzed using a shotgun proteomics approach based on filter-aided sample preparation followed by tandem mass spectrometry, protein identification carried out using Sequest-HT as the search engine within the Proteome Discoverer informatics platform, and label-free differential analysis. The dataset has been deposited in the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PRIDE: PXD004200.

Proteomic changes occurring along gonad maturation in the edible sea urchin Paracentrotus lividus.

UNLABELLED: The reproductive stage of Paracentrotus lividus strongly influences product quality that, in turn, impacts significantly on the market price. Large, compact and sweet gonads are preferred, and sensory attributes are positively related to the ratio of nutritive phagocytes to gametes. Gonads at advanced maturation stages, although larger, have less desirable attributes, being more watery and bitter especially in females. Therefore, the best compromise among size, texture, and taste needs to be reached. In this study, wild P. lividus were collected along coastal Sardinia, and gonads in the recovery, pre-mature, mature, and spent stages were analyzed by gel-based and by shotgun proteomics. A detailed characterization of the proteome changes occurring in gonads of both sexes along maturation was achieved, and significant modifications were seen in numerous proteins involved in nutrient accumulation in nutritive phagocytes, as well as in gamete biology and maturation. Adding to an improved understanding of the P. lividus reproductive cycle in its natural environment, the results described in this work may form the basis for defining novel protein markers and procedures for an easier sexing and staging, and for monitoring sea urchin gonad maturation cycles in aquaculture plants. Mass spectrometry data are deposited in ProteomeXchange (PXD004200). SIGNIFICANCE: The sensory quality of P. lividus gonads is strongly influenced by the reproductive cycle, with significant changes in flavor, texture, and size. A better knowledge of the protein profiles, patterns, and markers associated with gonad sex and maturation stage can have useful implications for understanding and monitoring these changes. One of these is the ability to identify protein profiles specifically associated with a given stage and, in perspective, to identify maturation and sex markers. The comprehensive proteomic evaluation achieved in this work was made possible by the application of combined gel-based and shotgun approaches. As a result, this study generated the largest proteomic dataset available in the literature for P. lividus, as well as a general picture of protein abundance changes occurring along maturation.

Influence of seasonal and environmental patterns on the lipid content and fatty acid profiles in gonads of the edible sea urchin Paracentrotus lividus from Sardinia.

The influence of seasonal and environmental patterns on the lipid fraction of Paracentrotus lividus gonads was investigated. For this purpose, sea urchins were collected monthly over a year from two Sardinian coastal areas. Total lipids in gonads follow an annual cyclical trend, described by a sine wave curve, that it is more influenced by season than by growing area. The lowest lipid content in gonads corresponds to a high percentage of mature reproductive stages (i.e. winter season), independently of sampling area. A variation in total lipid content follows a change in photoperiod, while it is related to sea surface temperature. Multivariate analysis on fatty acid profiles of gonads, detected by gas chromatography, clusters the collected specimens mainly according to the sampling area, secondly according to the sites within the same sampling area and finally according to season.

Impact of three commercial feed formulations on farmed gilthead sea bream (Sparus aurata, L.) metabolism as inferred from liver and blood serum proteomics.

BACKGROUND: The zootechnical performance of three different commercial feeds and their impact on liver and serum proteins of gilthead sea bream (Sparus aurata, L.) were assessed in a 12 week feeding trial. The three feeds, named A, B, and C, were subjected to lipid and protein characterization by gas chromatography (GC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), respectively. RESULTS: Feed B was higher in fish-derived lipids and proteins, while feeds C and A were higher in vegetable components, although the largest proportion of feed C proteins was represented by pig hemoglobin. According to biometric measurements, the feeds had significantly different impacts on fish growth, producing a higher average weight gain and a lower liver somatic index in feed B over feeds A and C, respectively. 2D DIGE/MS analysis of liver tissue and Ingenuity pathways analysis (IPA) highlighted differential changes in proteins involved in key metabolic pathways of liver, spanning carbohydrate, lipid, protein, and oxidative metabolism. In addition, serum proteomics revealed interesting changes in apolipoproteins, transferrin, warm temperature acclimation-related 65 kDa protein (Wap65), fibrinogen, F-type lectin, and alpha-1-antitrypsin. CONCLUSIONS: This study highlights the contribution of proteomics for understanding and improving the metabolic compatibility of feeds for marine aquaculture, and opens new perspectives for its monitoring with serological tests.

Proteomic profiling of sea bass muscle by two-dimensional gel electrophoresis and tandem mass spectrometry.

In this study, the proteome profile of European sea bass (Dicentrarchus labrax) muscle was analyzed using two-dimensional electrophoresis (2-DE) and tandem mass spectrometry with the aim of providing a more detailed characterization of its specific protein expression profile. A highly populated and well-resolved 2-DE map of the sea bass muscle tissue was generated, and the corresponding protein identity was provided for a total of 49 abundant protein spots. Upon Ingenuity Pathway Analysis, the proteins mapped in the sea bass muscle profile were mostly related to glycolysis and to the muscle myofibril structure, together with other biological activities crucial to fish muscle metabolism and contraction, and therefore to fish locomotor performance. The data presented in this work provide important and novel information on the sea bass muscle tissue-specific protein expression, which can be useful for future studies aimed to improve seafood traceability, food safety/risk management and authentication analysis. This work is also important for understanding the proteome map of the sea bass toward establishing the animal as a potential model for muscular studies.

Multidisciplinary analytical investigation of phospholipids and triglycerides in offshore farmed gilthead sea bream (Sparus aurata) fed commercial diets.

In this work, a quantitative characterisation of lipid (both triglycerides and phospholipids) rearrangements in the muscle of offshore-raised gilthead sea bream was carried out as a function of fish growth between April and September. Relative percentages of lipid classes and fatty acids/acyls composition of the commercial feeds and fish dorsal muscles were assessed by means of an interdisciplinary analytical approach. A combination of preparative chemistry and experimental results from NMR spectroscopy, GC, 3D-TLC as well as proximate analysis permitted the observed growth parameters in key metabolic events to be linked with fish fattening and lipid turnover. While defined effects of feed composition on fatty acid profiles of fillets were ascertained, the relative increase of fatty acyls in triglycerides and phospholipids were also estimated enabling detailed evaluation of TAG:PL ratio in adult offshore-farmed gilthead sea bream. NMR was also used to quantify PUFA regiospecific distribution in TAG and PL.

2D DIGE/MS to investigate the impact of slaughtering techniques on postmortem integrity of fish filet proteins.

Two-dimensional difference gel electrophoresis (2D DIGE) was applied to investigate the impact of slaughtering on the postmortem integrity of muscle tissue proteins in European sea bass (Dicentrarchus labrax). Three different slaughtering techniques were evaluated: asphyxia in air (AA), asphyxia in ice (AI), and spinal cord severance (SCS). Principal components analysis (PCA) revealed a significant divergence of SCS samples, whereas AA and AI samples, although grouped separately, were less divergent and could be included in a single asphyxia cluster. In terms of single proteins, the most significant impact was seen on nucleoside diphosphate kinase B, which was consistently less affected when fish were slaughtered by SCS as compared to asphyxia. Integrity of the sarcomeric proteins myosin heavy chain and myosin binding protein C and of the cytosolic proteins fructose biphosphate aldolase, glyceraldehyde 3-phosphate dehydrogenase, and enolase 1 was also better preserved upon SCS slaughtering. Most interestingly, the influence on muscle protein integrity could be detected since the early postmortem phase. In conclusion, slaughtering by SCS preserves protein integrity better than death by asphyxia, either in ice or in air. Both asphyxia conditions are comparably more adverse than SCS to muscle protein integrity, although a general trend favoring AI over AA is observed.

Bioconversion of ovine scotta into lactic acid with pure and mixed cultures of lactic acid bacteria.

Scotta is the main by-product in the making of ricotta cheese. It is widely produced in southern Europe and particularly in Italy where it represents a serious environmental pollutant due to its high lactose content. With the aim of evaluating whether scotta bioconversion into lactic acid can be considered as an alternative to its disposal, besides providing it with an added value, here the growth, fermentative performances, and lactic acid productions of pure and mixed cultures of Lactobacillus casei, Lactobacillus helveticus, and Streptococcus thermophilus were evaluated on ovine scotta-based media, without and with the addition of nutritional supplements. The outcomes indicate that ovine scotta can be utilized for the biotechnological production of lactic acid with yields up to 92%, comparable to those obtained on cheese-whey. Indeed, the addition of nutritional supplements generally improves the fermentative performances of lactic acid bacteria leading to about 2 g l(-1) h(-1) of lactic acid. Moreover, the use of mixed cultures for scotta bioconversion reduces the need for nutritional supplements, with no detrimental effects on the productive parameters compared to pure cultures. Finally, by using L. casei and S. thermophilus in pure and mixed cultures, up to 99% optically pure L: -lactic acid can be obtained.

Comparison of blood serum peptide enrichment methods by Tricine SDS-PAGE and mass spectrometry.

Characterisation of blood serum peptides can provide valuable information on physiological and pathological processes. However, the analysis of raw serum samples by MS results in the identification of a limited number of peptides. In order to improve sensitivity, many peptide enrichment methods have been proposed during the last ten years. Here, we present a comparison of fractionation methods aimed to simplify analysis of small proteins and peptides in blood serum, one of the most promising sources of putative biomarkers. Specifically, three methods based on ultrafiltration, differential precipitation, and peptide ligand libraries (ProteoMiner) were evaluated for the enrichment of peptides and low molecular weight proteins, as demonstrated by Tricine SDS-PAGE and subsequent LC-MS/MS (GeLC-MS/MS). As a result, differential solubilisation (DS) allowed the identification of the highest number of peptides. Moreover, the DS method enabled also the quantitative comparison of samples, producing fundamental information in biomarker discovery approaches.

Effects of postmortem storage temperature on sea bass (Dicentrarchus labrax) muscle protein degradation: analysis by 2-D DIGE and MS.

Storage conditions are known to be important for postmortem deterioration of fish muscle, and temperature is one of the factors with the strongest impact on this process. In order to shed light on the influence of temperature on the status of sea bass (Dicentrarchus labrax) muscle proteins during postmortem storage, a 2-D DIGE and mass spectrometry study was performed on fish kept at either 1 or 18°C for 5 days. As expected, the greatest alterations in sea bass filet protein composition were observed upon postmortem storage at 18°C, with distinct changes appearing in the 2-D protein profile after 5 days of storage at this temperature. In particular, degradation of the myofibrillar protein myosin heavy chain and of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase, among the most abundant muscle proteins, could be clearly observed upon storage at higher temperatures. Although to a lesser extent, however, several proteins were observed to vary in abundance also upon storage for 5 days at 1°C. In particular, one of the most interesting observations was the rapid and significant decrease in the abundance of nucleoside diphosphate kinase B and phosphoglycerate mutase 2, which was observed also at low storage temperatures and appeared to be temperature-independent. The results of this study offer new knowledge on changes occurring in sea bass muscle proteins during postmortem storage at different temperatures and provide indications on protein degradation trends that might be useful for monitoring freshness of fish and quality of storage conditions.

Proteomics and pathway analyses of the milk fat globule in sheep naturally infected by Mycoplasma agalactiae provide indications of the in vivo response of the mammary epithelium to bacterial infection.

Milk fat globules (MFGs) are vesicles released in milk as fat droplets surrounded by the endoplasmic reticulum and apical cell membranes. During formation and apocrine secretion by lactocytes, various amounts of cytoplasmic crescents remain trapped within the released vesicle, making MFGs a natural sampling mechanism of the lactating cell contents. With the aim of investigating the events occurring in the mammary epithelium during bacterial infection, the MFG proteome was characterized by two-dimensional difference gel electrophoresis (2-D DIGE), SDS-PAGE followed by shotgun liquid chromatography-tandem mass spectrometry (GeLC-MS/MS), label-free quantification by the normalized spectral abundance factor (NSAF) approach, Western blotting, and pathway analysis, using sheep naturally infected by Mycoplasma agalactiae. A number of protein classes were found to increase in MFGs upon infection, including proteins involved in inflammation and host defense, cortical cytoskeleton proteins, heat shock proteins, and proteins related to oxidative stress. Conversely, a strikingly lower abundance was observed for proteins devoted to MFG metabolism and secretion. To our knowledge, this is the first report describing proteomic changes occurring in MFGs during sheep infectious mastitis. The results presented here offer new insights into the in vivo response of mammary epithelial cells to bacterial infection and open the way to the discovery of protein biomarkers for monitoring clinical and subclinical mastitis.

Effect of modified atmosphere packaging on Quality Index Method (QIM) scores of farmed gilthead seabream (Sparus aurata L.) at low and abused temperatures.

A Quality Index Method (QIM) scheme was developed for modified atmosphere packaging (MAP) packed gilthead seabream, and the effect of MAP gas mixtures (60% CO2 and 40% N2; 60% CO2, 30% O2, and 10% N2), temperature (2, 4, and 8 °C), and time of storage on QI scores was assessed. QI scores were crossed with sensory evaluation of cooked fish according to a modified Torry scheme to establish the rejection point. In order to reduce redundant parameters, a principal component analysis was applied on preliminary QIM parameters scores coming from the best performing MAP among those tested. The final QIM scheme consists of 13 parameters and a maximum demerit score of 25. The maximum storage time was found to be 13 d at 4 °C for MAP 60% CO2 and 40% N2. Storage at 2 °C do not substantially improved sensory parameters scores, while storage under temperature abuse (8 °C) accelerated drastically the rate of increase of QI scores and reduced the maximum storage time to 6 d.

Application of 2-D DIGE to formalin-fixed, paraffin-embedded tissues.

The ability to investigate the proteome of formalin-fixed, paraffin-embedded (FFPE) tissues can be considered a major recent achievement in the field of clinical proteomics. However, gel-based approaches to the investigation of FFPE tissue proteomes have lagged behind, mainly because of insufficient quality of full-length protein extracts. Here, the 2-D DIGE technology was investigated for applicability to FFPE proteins, for internal reproducibility among replicate FFPE extracts, and for comparability between FFPE and fresh-frozen tissue profiles. The 2-D DIGE patterns obtained upon labeling and electrophoresis of replicate FFPE tissue extracts were highly reproducible, with satisfactory resolution and complexity. Moreover, the implementation of DIGE enabled to highlight and characterize the consistent differences found in the FFPE profiles compared with fresh-frozen profiles, represented by an acidic shift, directly correlated to the protein pI value, and by a reduction in spot signal intensity, directly correlated to molecular weight and percentage of lysine residues. Being constantly and reproducibly present in all FFPE tissue extract replicates at similar extents, these modifications do not appear to hinder the comparative analysis of FFPE tissue extracts by 2-D DIGE, opening the way to its application for the differential proteomic investigation of archival tissue repositories.