Effect of processing on in vitro digestibility (IVPD) of food proteins.

Proteins are important macronutrients for the human body to grow and function throughout life. Although proteins are found in most foods, their very dissimilar digestibility must be taking into consideration when addressing the nutritional composition of a diet. This review presents a comprehensive summary of the in vitro digestibility of proteins from plants, milk, muscle, and egg. It is evident from this work that protein digestibility greatly varies among foods, this variability being dependent not only upon the protein source, but also the food matrix and the molecular interactions between proteins and other food components (food formulation), as well as the conditions during food processing and storage. Different approaches have been applied to assess in vitro protein digestibility (IVPD), varying in both the enzyme assay and quantification method used. In general, animal proteins tend to show higher IVPD. Harsh technological treatments tend to reduce IVPD, except for plant proteins, in which thermal degradation of anti-nutritional compounds results in improved IVPD. However, in order to improve the current knowledge about protein digestibility there is a vital need for understanding dependency on a protein source, molecular interaction, processing and formulation and relationships between. Such knowledge can be used to develop new food products with enhanced protein bioaccessibility.

Development of Volatile Compounds during Hydrolysis of Porcine Hemoglobin with Papain.

There is a growing market for the use of hydrolysates from animal side-streams for production of high-protein supplements. However, there can be issues with development of off-flavors, either due to the raw material in question or due to the hydrolysis process itself. This study examined the development of volatile compounds during hydrolysis of hemoglobin. Briefly, porcine hemoglobin was hydrolyzed by 0.5% papain for up to 5 h, and the development of volatile compounds was analyzed via gas chromatography-mass spectrometry. The results showed that there was significant development of a number of volatile compounds with time, e.g., certain Maillard reaction and lipid oxidation products, which are likely candidates for the aroma development during hydrolysis. Furthermore, it was shown that development of a number of the volatiles was due to the hydrolysis process, as these compounds were not found in a control without enzyme.

Improving the lipid profile of ready-to-cook meat products by addition of omega-3 microcapsules: effect on oxidation and sensory analysis.

BACKGROUND: The omega-3 enrichment of ready-to-cook meat products by microencapsulated fish oil (MFO) addition was analyzed. Accordingly, three batches of chicken nuggets were prepared: (i) control (C); (ii) enriched in bulk fish oil (BFO); and (iii) with added MFO. Sensory features, acceptability, oxidative stability and volatile compounds were analyzed. RESULTS: MFO nuggets did not differ from C ones with respect to any sensory trait. BFO showed increased juiciness and saltiness but decreased meat flavor. Acceptability was not affected by enrichment. Consumers were not able to differentiate between C and MFO in a triangle test, although they could clearly identify BFO nuggets. Higher levels of lipid and protein oxidation indicators and of volatile compounds from fatty acid oxidation were found in BFO nuggets compared to C and MFO nuggets. CONCLUSION: Enrichment of ready-to-cook meat products in omega-3 fatty acids with MFO provides both lipid and protein oxidative protection without changes in sensory quality. © 2018 Society of Chemical Industry.

Investigation of nitrite alternatives for the color stabilization of heme-iron hydrolysates.

This study investigates the potential of novel heme-ligand complexes, derived from heme-iron isolated from porcine hemoglobin by enzymatic hydrolysis, to use as pigments for meat products. Five alternatives to sodium nitrite were identified as possible heme ligands and stabilizing agents of the red conformation of heme. The effects of 4-methylimidazole, methyl nicotinate, pyrrolidine, piperidine, pyrazine and sodium nitrite (as comparative benchmark) on the color of heme-iron extract and pure hemin standard were studied in solution. The ligand affinity and heme-ligand stability was assessed over time in solution by UV-Vis absorbance spectroscopy and CIELAB color space parameters. The CIE redness score a* was used as a single measurement to propose a predictive model based on the following parameters: heme source (heme-iron extract or hemin standard), heme-to-ligand molar ratio (1:20 to 1:300), and storage time (up to 32 days). The optimal concentration at which each ligand can be added to either heme source, as well as the stability of the red color of the formed heme-ligand complexes in-solution was determined. Heme-iron extract-derived samples showed increased redness and color stability as compared to their hemin counterparts. No ligand showed as much affinity for heme as sodium nitrite. As the most promising ligand candidates, methyl nicotinate and 4-methylimidazole started to show color changes at a 1:50 molar ratio, but higher amounts (1:100 and 1:300, respectively) were required to attain the maximum redness possible with the highest stability.

B-cell anergy induces a Th17 shift in a novel B lymphocyte transgenic NOD mouse model, the 116C-NOD mouse.

Autoreactive B lymphocytes play a key role as APCs in diaebetogenesis. However, it remains unclear whether B-cell tolerance is compromised in NOD mice. Here, we describe a new B lymphocyte transgenic NOD mouse model, the 116C-NOD mouse, where the transgenes derive from an islet-infiltrating B lymphocyte of a (8.3-NODxNOR) F1 mouse. The 116C-NOD mouse produces clonal B lymphocytes with pancreatic islet beta cell specificity. The incidence of T1D in 116C-NOD mice is decreased in both genders when compared with NOD mice. Moreover, several immune selection mechanisms (including clonal deletion and anergy) acting on the development, phenotype, and function of autoreactive B lymphocytes during T1D development have been identified in the 116C-NOD mouse. Surprisingly, a more accurate analysis revealed that, despite their anergic phenotype, 116C B cells express some costimulatory molecules after activation, and induce a T-cell shift toward a Th17 phenotype. Furthermore, this shift on T lymphocytes seems to occur not only when both T and B cells contact, but also when helper T (Th) lineage is established. The 116C-NOD mouse model could be useful to elucidate the mechanisms involved in the generation of Th-cell lineages.

Oxidative deterioration of pork during superchilling storage.

BACKGROUND: In superchilling (SC), meat is kept at temperatures around 1 °C below its initial freezing point, leading to a significant increase in shelf life. This study aimed to address the oxidative changes taking place in pork loins during prolonged storage at SC temperature. Loins were stored either at chilling (CH) conditions (2-4 °C) for 4 weeks or at SC temperature (around -1 °C) for 12 weeks. RESULTS: Storage at SC temperature diminished the rate of lipid and protein oxidation and discoloration in pork loins, so that final levels of most oxidation products and instrumental color values after 12 weeks of SC storage were similar to those after 4 weeks at CH conditions. However, hexanal content peaked by the end of SC storage, pointing to a potential accumulation of compounds from lipid oxidation during SC storage. CONCLUSION: SC storage of pork slows down the rate of lipid and protein oxidation. However, accumulation of volatile compounds from lipid oxidation could be a limiting factor for shelf life. © 2017 Society of Chemical Industry.

In vivo detection of peripherin-specific autoreactive B cells during type 1 diabetes pathogenesis.

Autoreactive B cells are essential for the pathogenesis of type 1 diabetes. The genesis and dynamics of autoreactive B cells remain unknown. In this study, we analyzed the immune response in the NOD mouse model to the neuronal protein peripherin (PRPH), a target Ag of islet-infiltrating B cells. PRPH autoreactive B cells recognized a single linear epitope of this protein, in contrast to the multiple epitope recognition commonly observed during autoreactive B cell responses. Autoantibodies to this epitope were also detected in the disease-resistant NOR and C57BL/6 strains. To specifically detect the accumulation of these B cells, we developed a novel approach, octameric peptide display, to follow the dynamics and localization of anti-PRPH B cells during disease progression. Before extended insulitis was established, anti-PRPH B cells preferentially accumulated in the peritoneum. Anti-PRPH B cells were likewise detected in C57BL/6 mice, albeit at lower frequencies. As disease unfolded in NOD mice, anti-PRPH B cells invaded the islets and increased in number at the peritoneum of diabetic but not prediabetic mice. Isotype-switched B cells were only detected in the peritoneum. Anti-PRPH B cells represent a heterogeneous population composed of both B1 and B2 subsets. In the spleen, anti-PRPH B cell were predominantly in the follicular subset. Therefore, anti-PRPH B cells represent a heterogeneous population that is generated early in life but proliferates as diabetes is established. These findings on the temporal and spatial progression of autoreactive B cells should be relevant for our understanding of B cell function in diabetes pathogenesis.

Volatile compounds profile of sous-vide cooked pork cheeks as affected by cooking conditions (vacuum packaging, temperature and time).

The volatile organic compound (VOC) profile of pork cheeks as affected by the cooking conditions was investigated. Pork cheeks were cooked under different combinations of temperature (60 °C or 80 °C), time (5 or 12 h) and vacuum (vacuum or air-packaged). As a general rule, the VOCs originating from lipid degradation were positively affected by the cooking temperature and negatively by the cooking time, reaching the highest amounts in pork cheeks cooked at 80 °C during 5 h and the lowest in samples cooked at 80 °C during 12 h. On the contrary, VOCs originated from amino acids and Maillard reactions were positively affected by both factors. The proportion between lipid degradation and amino acids reactions was estimated by the hexanal/3-methylbutanal ratio, which reached its highest values in samples cooked at 60 °C during 5 h in the presence of air and the lowest values in samples cooked at 80 °C during 12 h, regardless of the vacuum status.

Cooking affects pork proteins in vitro rate of digestion due to different structural and chemical modifications.

The effect of thermal processing on the in vitro digestibility of pork proteins was studied. Raw samples were considered the control group, while the thermal treatments included 58, 80, 98 and 160 °C for 72 min, 118 °C for 8 min and 58 °C for 17 h, resembling a range of different cooking procedures. Samples were subsequently subjected to pepsin digestion at pH 3.00 in the gastric phase followed by trypsin and α-chymotrypsin at pH 8.00 in the intestinal phase. Pork cooked at 58 °C for 72 min had a significantly higher pepsin digestibility rate than meat cooked at 80 °C or 160 °C. The trend was similar in the intestinal phase, with samples cooked at 58 °C for 72 min having enhanced digestion rate over other treatments after 120 min of digestion. A PLS model pointed out to an inverse relationship between in vitro proteolysis rate and variables like Maillard reaction compounds or protein structural changes.

Improvements in the methodology for fatty acids analysis in meat products: One-stage transmethylation and fast-GC method.

The quantification of fatty acids (FA) in meat products is frequently carried out by two-stage methylation procedures followed by long gas chromatography (GC) runs. This work aimed to simplify this methodology by means of a one-stage transmethylation method and a fast GC run, evaluating the influence of sample preparation, reagents and type of heating on the amount of FA in different meat products and optimizing a fast GC-FID (flame ionization detector) run. This allowed to establish the optimum combination of parameters (methanol + chlorotrimethylsilane, lyophilized samples and oven heating) to achieve the quantification of the highest possible amount of FA and to reduce the time of GC run from 60 to 10 min. The quality evaluation of this method obtained satisfactory results. Thus, the quantification of FA in meat products was achieved in a straightforwardly and quickly way by using a one-stage transmethylation procedure followed by a fast GC-FID run.

Recent Developments in the Microencapsulation of Fish Oil and Natural Extracts: Procedure, Quality Evaluation and Food Enrichment.

Due to the beneficial health effects of omega-3 fatty acids and antioxidants and their limited stability in response to environmental and processing factors, there is an increasing interest in microencapsulating them to improve their stability. However, despite recent developments in the field, no specific review focusing on these topics has been published in the last few years. This work aimed to review the most recent developments in the microencapsulation of fish oil and natural antioxidant compounds. The impact of the wall material and the procedures on the quality of the microencapsulates were preferably evaluated, while their addition to foods has only been studied in a few works. The homogenization technique, the wall-material ratio and the microencapsulation technique were also extensively studied. Microcapsules were mainly analyzed for size, microencapsulation efficiency, morphology and moisture, while in vitro digestion, flowing properties, yield percentage and Fourier transform infrared spectroscopy (FTIR) were used more sparingly. Findings highlighted the importance of optimizing the most influential variables of the microencapsulation procedure. Further studies should focus on extending the range of analytical techniques upon which the optimization of microcapsules is based and on addressing the consequences of the addition of microcapsules to food products.

Meat and Human Health-Current Knowledge and Research Gaps.

Meat is highly nutritious and contributes with several essential nutrients which are difficult to obtain in the right amounts from other food sources. Industrially processed meat contains preservatives including salts, possibly exerting negative effects on health. During maturation, some processed meat products develop a specific microbiota, forming probiotic metabolites with physiological and biological effects yet unidentified, while the concentration of nutrients also increases. Meat is a source of saturated fatty acids, and current WHO nutrition recommendations advise limiting saturated fat to less than ten percent of total energy consumption. Recent meta-analyses of both observational and randomized controlled trials do not support any effect of saturated fat on cardiovascular disease or diabetes. The current evidence regarding the effect of meat consumption on health is potentially confounded, and there is a need for sufficiently powered high-quality trials assessing the health effects of meat consumption. Future studies should include biomarkers of meat intake, identify metabolic pathways and include detailed study of fermented and other processed meats and their potential of increasing nutrient availability and metabolic effects of compounds.

Flavor Characterization of Animal Hydrolysates and Potential of Glucosamine in Flavor Modulation.

Bovine (meat and heart) and porcine (hemoglobin and plasma) raw materials were hydrolyzed by Protease A (both endo- and exopeptidase activity), with or without glucosamine added during the enzyme inactivation step. Hydrolysates were characterized via peptide analysis (yield, UV- and fluorescence scanning spectroscopy, and peptide size distribution via size exclusion chromatography), sensory evaluation, and volatile compound analysis via gas chromatography mass-spectrometry (GC-MS) to determine if glucosamine-induced Maillard reaction improved taste and flavor. Porcine hemoglobin produced the most flavor-neutral hydrolysate, and could expectedly have the broadest application in food products. Both bovine meat and -heart hydrolysates were high in umami, and thereby good candidates for savory applications. Porcine plasma hydrolysate was high in liver flavor and would be suitable for addition to certain meat products where liver flavor is desirable. All hydrolysates had low perceived bitterness. Glucosamine-induced Maillard reaction had just a minor influence on the sensory profile via an increased perception of sweet taste (p = 0.038), umami taste (p = 0.042), and yolk flavor (p = 0.038) in the hydrolysates, irrespective of raw material. Glucosamine addition had a statistically significant effect on 13 of 69 volatiles detected in the hydrolysates, but the effect was minor and raw material-specific.

Monitoring the Processing of Dry Fermented Sausages with a Portable NIRS Device.

This work studies the ability of a MicroNIR (VIAVI, Santa Rosa, CA) device to monitor the dry fermented sausage process with the use of multivariate data analysis. Thirty sausages were made and subjected to dry fermentation, which was divided into four main stages. Physicochemical (weight lost, pH, moisture content, water activity, color, hardness, and thiobarbiruric reactive substances analysis) and sensory (quantitative descriptive analysis) characterizations of samples on different steps of the ripening process were performed. Near-infrared (NIR) spectra (950-1650 nm) were taken throughout the process at three points of the samples. Physicochemical data were explored by distance to K-Nearest Neighbor (K-NN) cluster analysis, while NIR spectra were studied by partial least square-discriminant analysis; before these models, Principal Component Analysis (PCA) was performed in both databases. The results of multivariate data analysis showed the ability to monitor and classify the different stages of ripening process (mainly the fermentation and drying steps). This study showed that a portable NIR device (MicroNIR) is a nondestructive, simple, noninvasive, fast, and cost-effective tool with the ability to monitor the dry fermented sausage processing and to classify samples as a function of the stage, constituting a feasible decision method for sausages to progress to the following processing stage.

Changes in the Quality of Chicken Breast Meat due to Superchilling and Temperature Fluctuations during Storage.

The aim of this study was to determine the changes in chicken breast meat quality (water-holding capacity, color, texture, myofibrillar fragmentation index (MFI), total protein solubility, thiobarbituric acid reactive substances (TBARS), total viable count (TVC), and lactic acid bacteria (LAB) count) due to storage under superchilling conditions (-1.3°C) and fluctuating temperatures (ranging from -20°C to -5°C) as compared to the quality of meat stored at chilled (2-4°C) and frozen (-20°C) temperatures, respectively. Results indicated that the TVC and LAB count of the chilled and superchilled breast meat increased with storage time. TVC of the chilled and superchilled breast meat reached the safety level of 7 log cfu/g at approximately day 8 and18, respectively. This suggested that the superchilling method extended the storage duration by 10 days. Weight loss and TBARS of the chilled and superchilled samples tended to increase with increasing storage time. The color, texture, protein solubility, and MFI were stable throughout the entire storage period of the chilled (9 days) and superchilled (28 days) samples. Results indicated that while three cycles of storage temperature fluctuation influenced the weight loss and dry matter of the meat, they did not affect the TVC, LAB count, texture, color, pH, MFI, and protein solubility. The superchilling technique (-1.3°C) could extend the shelf-life of meat and maintain the quality of chicken breast meat. Fluctuations in temperature during frozen storage decreased the water-holding capacity of chicken breast meat, indicating that temperature stability should be maintained during frozen storage.

B-Lymphocyte Phenotype Determines T-Lymphocyte Subset Differentiation in Autoimmune Diabetes.

Previous studies indicate that B-lymphocytes play a key role activating diabetogenic T-lymphocytes during the development of autoimmune diabetes. Recently, two transgenic NOD mouse models were generated: the NOD-PerIg and the 116C-NOD mice. In NOD-PerIg mice, B-lymphocytes acquire an activated proliferative phenotype and support accelerated autoimmune diabetes development. In contrast, in 116C-NOD mice, B-lymphocytes display an anergic-like phenotype delaying autoimmune diabetes onset and decreasing disease incidence. The present study further evaluates the T- and B-lymphocyte phenotype in both models. In islet-infiltrating B-lymphocytes (IIBLs) from 116C-NOD mice, the expression of H2-Kd and H2-Ag7 is decreased, whereas that of BAFF, BAFF-R, and TACI is increased. In contrast, IIBLs from NOD-PerIg show an increase in CD86 and FAS expression. In addition, islet-infiltrating T-lymphocytes (IITLs) from NOD-PerIg mice exhibit an increase in PD-1 expression. Moreover, proliferation assays indicate a high capacity of B-lymphocytes from NOD-PerIg mice to secrete high amounts of cytokines and induce T-lymphocyte activation compared to 116C B-lymphocytes. This functional variability between 116C and PerIg B-lymphocytes ultimately results in differences in the ability to shape T-lymphocyte phenotype. These results support the role of B-lymphocytes as key regulators of T-lymphocytes in autoimmune diabetes and provide essential information on the phenotypic characteristics of the T- and B-lymphocytes involved in the autoimmune response in autoimmune diabetes.

Protein hydrolysates of porcine hemoglobin and blood: Peptide characteristics in relation to taste attributes and formation of volatile compounds.

The objective of this study was to investigate the impact of endo- and exo-peptidase treatment on certain structural characteristics of peptides and volatile compounds of porcine hemoglobin and whole blood hydrolysates. Porcine hemoglobin and whole blood were hydrolyzed by endo- and exo-peptidases. The presence of exopeptidases reduced the bitterness and altered the volatile profiles of protein hydrolysates. Exopeptidase treatment can release terminal amino acids from peptides, which in turn may contribute to formation of volatile compounds by Maillard reactions. In contrast, endopeptidases conferred a slightly bitter taste and different volatile profiles. For hemoglobin hydrolysates, principal component analysis revealed that proteases were categorized into three groups based on endo- or exo-peptidase activity. Whole blood is a more complex raw material, yet the proteases were still categorized in a similar fashion. This work contributes to understanding structural characteristics responsible for taste and volatile profiles of protein hydrolysates.

Proteolysis in pork loins during superchilling and regular chilling storage.

This study aimed to address the proteolytic phenomena taking place in pork loins during prolonged storage at superchilling (SC) temperature. Loins were stored at either chilling (CH) conditions (2-4 °C) for 4 weeks or at SC temperature (around -1 °C) for 12 weeks. Storage at SC temperatures slowed down the rate of proteolysis in pork loins, so that final levels of most indicators for proteolysis, including after 12 weeks of SC storage were similar to those after 4 weeks at CH conditions. Consequently, the texture of SC pork under extended storage was not so different to that of CH pork. However, total amino acid content peaked by the end of SC storage, pointing out to a potential ongoing exopeptidase activity. Overall, proteolysis seemed to be slowed down in pork at SC conditions, with similar levels for most indicators after 12 weeks of SC storage or 4 weeks at CH conditions.

Pork proteins oxidative modifications under the influence of varied time-temperature thermal treatments: A chemical and redox proteomics assessment.

The impact of thermal processing on meat proteins oxidation was investigated. Cooking treatments included 58 °C for either 72 min or 17 h (mimicking low temperature-long time sous vide cooking), 80, 98 and 160 °C for 72 min (mimicking common cooked meat products, stewing and roasting, respectively) and 118 °C for 8 min (autoclaving). Tryptophan degradation, fluorescent oxidation products, free thiol content and α-aminoadipic and γ-glutamic semialdehydes were tracked. For all of them, there was a consistent trend to increased levels of oxidative damage with higher cooking temperatures and longer cooking times, although the extent varied from one indicator to another. Through proteomics, peptide oxidative modifications like carbonylation, malonaldehyde adducts and hydroxykynurenin (tryptophan oxidation products) were also detected at residue level. Our findings indicate that protein oxidation is dependent upon the heat treatment, which point out to a different effect on the nutritional quality of proteins in meat products.

Advanced glycation end products, protein crosslinks and post translational modifications in pork subjected to different heat treatments.

The aim of the study was to characterize Maillard reactions in meat under different cooking treatments. Considered temperature-time combinations included raw samples (control), 58, 80, 98 and 160 °C for 72 min, 118 °C for 8 min and 58 °C for 17 h. Furosine, a marker for heat treatment, was detected in all groups with roasting having a 4-fold increase over the control. Sous-vide treatment at 80 °C, boiling and autoclaving also contribute to a significant increase in furosine. Nɛ-carboxymethyllysine, an indicator for advanced glycation end products, showed negligible amount in control, but increased with cooking temperature, with oven samples showing the highest values. A similar increasing trend was observed in lanthionine, covalently bonded protein crosslinks, which arises due to severe thermal regimes. Simultaneously, glycation and deamidation formation were tracked in meat proteins through peptidomics to highlight residue level changes that might affect nutrient value in processed muscle based foods.