An integrated peptidomics and in silico approach to identify novel anti-diabetic peptides in parmigiano-reggiano cheese

Inhibition of key metabolic enzymes linked to type-2-diabetes (T2D) by food-derived compounds is a preventive emerging strategy in the management of T2D. Here, the impact of Parmigiano- Reggiano (PR) cheese peptide fractions, at four different ripening times (12, 18, 24, and 30 months), on the enzymatic activity of α-glucosidase, α-amylase, and dipeptidyl peptidase-IV (DPPIV) as well as on the formation of fluorescent advanced glycation end-products (fAGEs) was assessed. The PR peptide fractions were able to inhibit the selected enzymes and fAGEs formation. The 12-month-ripening PR sample was the most active against the three enzymes and fAGEs. Mass spectrometry analysis enabled the identification of 415 unique peptides, 54.9% of them common to the four PR samples. Forty-nine previously identified bioactive peptides were found, mostly characterized as angiotensin-converting enzyme-inhibitors. The application of an integrated approach that combined peptidomics, in silico analysis, and a structure–activity relationship led to an efficient selection of 6 peptides with potential DPP-IV and α-glucosidase inhibitory activities. Peptide APFPE was identified as a potent novel DPP-IV inhibitor (IC50 = 49.5 ± 0.5 μmol/L). In addition, the well-known anti-hypertensive tripeptide, IPP, was the only one able to inhibit the three digestive enzymes, highlighting its possible new and pivotal role in diabetes management

Exploring the potential of NIR hyperspectral imaging for automated quantification of rind amount in grated Parmigiano Reggiano cheese

Parmigiano Reggiano (P-R) is one of the most important Italian food products labelled with Protected Designation of Origin (PDO). The PDO denomination is applied also to grated P-R cheese products meeting the requirements regulated by the Specifications of Parmigiano Reggiano Cheese. Different quality parameters are monitored, including the percentage of rind, which is edible and should not exceed the limit of 18% (w/w). The present study aims at evaluating the possibility of using near infrared hyperspectral imaging (NIR-HSI) to quantify the rind percentage in grated Parmigiano Reggiano cheese samples in a fast and non-destructive manner. Indeed, NIR-HSI allows the simultaneous acquisition of both spatial and spectral information from a sample, which is more suitable than classical single-point spectroscopy for the analysis of heterogeneous samples like grated cheese. Hyperspectral images of grated P-R cheese samples containing increasing levels of rind were acquired in the 900–1700 nm spectral range. Each hyperspectral image was firstly converted into a one-dimensional signal, named hyperspectrogram, which codifies the relevant information contained in the image. Then, the matrix of hyperspectrograms was used to calculate a calibration model for the prediction of the rind percentage using Partial Least Squares (PLS) regression. The calibration model was validated considering two external test sets of samples, confirming the effectiveness of the proposed approach

Evaluation of the effect of factors related to preparation and composition of grated Parmigiano Reggiano cheese using NIR hyperspectral imaging

The present study is focused on the evaluation of the effect of grater type and fat content of the pulp on the spectral response obtained by near infrared hyperspectral imaging (NIR-HSI), when this technique is used to determine the rind percentage in Parmigiano Reggiano (P-R) cheese. To this aim, grated P-R cheese samples were prepared considering all the possible combinations between three levels of rind amount (8%, 18% and 28%), two levels of fat content of the pulp and two different grater types, and the corresponding hyperspectral images were acquired in the 900–1700 nm spectral range. In a first step, the average spectrum (AS) was calculated from each hyperspectral image, and the corresponding dataset was analysed by means of Analysis of Variance Simultaneous Component Analysis (ASCA) to assess the effect of the three considered factors and their two-way interactions on the spectral response. Then, the hyperspectral images were converted into Common Space Hyperspectrograms (CSH), which are signals obtained by merging in sequence the frequency distribution curves of quantities calculated from a Principal Component Analysis (PCA) model common to the whole hyperspectral image dataset. ASCA was also applied to the CSH dataset, in order to evaluate the effect of the considered factors on this kind of signals. Generally, all the three factors resulted to have a significant effect, but with a different extent according to the method used to analyse the hyperspectral images. Indeed, while fat content of the pulp and rind percentage showed a comparable effect on the spectral response of AS dataset, in the case of CSH signals rind percentage had a greater effect compared to the other main factors. However, CSH were also more sensitive to differences ascribable to the natural variability between diverse Parmigiano Reggiano cheese samples

Peptide profiling and biological activities of 12- month ripened parmigiano reggiano cheese

Proteolysis degree, biological activities, and water-soluble peptide patterns were evaluated in 12 month-ripened Parmigiano Reggiano (PR) cheeses collected in different dairy farms and showing different salt and fat content. Samples classified in high-salt and high-fat group (HH) generally showed lower proteolysis degree than samples having low-salt and low-fat content (LL). This positive correlation between salt/fat reduction and proteolysis was also confirmed by the analysis of biological activities, as the LL group showed higher average values of angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. UHPLC/HR-MS allowed the identification of 805 unique peptides: LL and HH groups shared 59.3% of these peptides, while 20.9% and 19.9% were LL and HH specific, respectively. Frequency analysis of peptides identified a core of 183 peptides typical of 12-month ripened PR cheeses (corresponding to the 22.7% of total peptides), but no significant differences were detected in peptide patterns between LL and HH groups. Forty bioactive peptides, including 18 ACE-inhibitors and 12 anti-microbial peptides, were identified, of which 25 firstly found in PR cheese. Globally, this work contributed to unraveling the potentially healthy benefits of peptides fraction in PR cheese and provided prior evidence that PR with reduced at/salt content showed the highest antihypertensive and antioxidant activities

Characterization of yeasts isolated from parmigiano reggiano cheese natural whey starter: From spoilage agents to potential cell factories for whey valorization

Whey is the main byproduct of the dairy industry and contains sugars (lactose) and proteins (especially serum proteins and, at lesser extent, residual caseins), which can be valorized by the fermentative action of yeasts. In the present study, we characterized the spoilage yeast population inhabiting natural whey starter (NWS), the undefined starter culture of thermophilic lactic acid bacteria used in Parmigiano Reggiano (PR) cheesemaking, and evaluated thermotolerance, mating type, and the aptitude to produce ethanol and bioactive peptides from whey lactose and proteins, respectively, in a selected pool of strains. PCR-RFLP assay of ribosomal ITS regions and phylogenetic analysis of 26S rDNA D1/D2 domains showed that PR NWS yeast population consists of the well-documented Kluyveromyces marxianus, as well as of other species (Saccharomyces cerevisiae, Wickerhamiella pararugosa, and Torulaspora delbrueckii), with multiple biotypes scored within each species as demonstrated by (GTG)5-based MSP-PCR. Haploid and diploid K. marxianus strains were identified through MAT genotyping, while thermotolerance assay allowed the selection of strains suitable to grow up to 48◦C. In whey fermentation trials, one thermotolerant strain was suitable to release ethanol with a fermentation efficiency of 86.5%, while another candidate was able to produce the highest amounts of both ethanol and bioactive peptides with potentially anti-hypertensive function. The present work demonstrated that PR NWS is a reservoir of ethanol and bioactive peptides producer yeasts, which can be exploited to valorize whey, in agreement with the principles of circularity and sustainability

Is right angular gyrus involved in the metric component of the mental body representation in touch and vision? A tdcs study

Several studies have found in the sense of touch a good sensory modality by which to study body representation. Here, we address the “metric component of body representation”, a specific function developed to process the discrimination of tactile distances on the body. The literature suggests the involvement of the right angular gyrus (rAG) in processing the tactile metricity on the body. The question of this study is the following: is the rAG also responsible for the visual metric component of body representation? We used tDCS (anodal and sham) in 20 subjects who were administered an on-body distance discrimination task with both tactile and visual stimuli. They were also asked to perform the same task in a near-body condition. The results allow us to confirm the role of rAG in the estimation of tactile distances. Further, we also showed that rAG might be involved in the discrimination of distances on the body not only in tactile but also in visual modality. Finally, based on the significant effects of anodal stimulation even in a near-body visual discrimination task, we proposed a higher-order function of the AG in terms of a supramodal com-parator of quantities

Tracing the identity of Parmigiano Reggiano “Prodotto di Montagna - Progetto Territorio” cheese using NMR spectroscopy and multivariate data analysis

Background Nuclear magnetic resonance (NMR) spectroscopy is one of the well-established tools for food metabolomic analysis, as it proved to be very effective in authenticity and quality control of dairy products, as well as to follow product evolution during processing and storage. The analytical assessment of the EU mountain denomination label, specifically for Parmigiano Reggiano "Prodotto di Montagna - Progetto Territorio" (Mountain-CQ) cheese, has received limited attention. Although it was established in 2012 the EU mountain denomination label has not been much studied from an analytical point of view. Nonetheless, tracing a specific profile for the mountain products is essential to support the value chain of this specialty. Results The aim of the study was to produce an identity profile for Parmigiano Reggiano “Prodotto di Montagna - Progetto Territorio” (Mountain-CQ) cheese, and to differentiate it from Parmigiano Reggiano PDO samples (conventional-PDO) using 1H NMR spectroscopy coupled with multivariate data analysis. Three different approaches were applied and compared. First, the spectra-as-such were analysed after proper preprocessing. For the other two approaches, Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) was used for signals resolution and features extraction, either individually on manually-defined spectral intervals or by reapplying MCR-ALS on the whole spectra with selectivity constraints using the reconstructed “pure profiles” as initial estimates and targets. All approaches provided comparable information regarding the samples’ distribution, as in all three cases the separation between the two product categories conventional-PDO and Mountain-CQ could be highlighted. Moreover, a novel MATLAB toolbox for features extraction via MCR-ALS was developed and used in synergy with the Chenomx library, allowing for a putative identification of the selected features. Significance A first identity profile for Parmigiano Reggiano “Prodotto di Montagna - Progetto Territorio” obtained by interpreting the metabolites signals in NMR spectroscopy was obtained. Our workflow and toolbox for generating the features dataset allows a more straightforward interpretation of the results, to overcome the limitations due to dimensionality and to peaks overlapping, but also to include the signals assignment and matching since the early stages of the data processing and analysis

Factors Affecting the Clinical Measurement of Visuo-Spatial Neglect

The present study examined a battery of tests to evaluate unilateral spatial neglect; the tests included different tasks involving several modalities of spatial exploration mapping perceptual, motor, attentional and personal or extrapersonal space dimensions. The subjects, 121 right-brain-damaged patients with unilateral neglect, were studied in seven laboratories in four European countries. Relationships among the various tests were examined by correlations, a cluster analysis and by an analysis of individual cases. Different sensitivity was found among various tests for detecting neglect performances. Both the cluster analysis and the single case analysis clearly showed a segregation between personal and extrapersonal neglect. Analysis of the large cluster, including a variety of tests of extra personal neglect, together with the study of single cases, suggests the possibility of differentiating the various manifestations of spatial neglect which can be interpreted on the basis of the descriptions of other individual cases previously reported in the literature. Finally, the present study indicated the relative stability of neglect following the acute phase and its independence from age

Identifying the cognitive processes underpinning hippocampal-dependent tasks

Autobiographical memory, future thinking, and spatial navigation are critical cognitive functions that are thought to be related and are known to depend upon a brain structure called the hippocampus. Surprisingly, direct evidence for their interrelatedness is lacking, as is an understanding of why they might be related. There is debate about whether they are linked by an underlying memory-related process or, as has more recently been suggested, because they each require the endogenous construction of scene imagery. Here, using a large sample of participants and multiple cognitive tests with a wide spread of individual differences in performance, we found that these functions are indeed related. Mediation analyses further showed that scene construction, and not memory, mediated (explained) the relationships between the functions. These findings offer a fresh perspective on autobiographical memory, future thinking, navigation, and also on the hippocampus, where scene imagery appears to play an influential role