Influence of enzymatic hydrolysis and molecular weight fractionation on the antioxidant and lipase / α-amylase inhibitory activities in vitro of watermelon seed protein hydrolysates

This study aims to evaluate the potential in vitro antioxidant and anti-obesity activities of watermelon seed protein hydrolysates (WSPH) obtained using different combinations of enzymes alcalase–proteinase K (ALC-PK) and alcalase–actinidin (ALC-ACT). There was a direct relationship between the degree of hydrolysis (DH) and the biological activities of the WSPH, with the highest DPPH (approximately 85%) and lipase inhibitory activities (≈59%) appreciated at DH of 36–37% and 33–35% when using ALC-PK and ALC-ACT, respectively. Following molecular weight fractionation, the ALC-PK WSPH < 3 kDa (F1) assayed at 1 mg.mL−1 had the highest DPPH-radical scavenging (89.22%), ferrous chelating (FC) (79.83%), reducing power (RP) (A 0.51), lipase inhibitory (71.36%), and α-amylase inhibitory (62.08%) activities. The amino acid analysis of ALC-PK WSPH and its fractions revealed a relationship between the biological activity of the extracts and their composition. High contents of hydrophobic amino acids, arginine, and aromatic amino acids were related to high antioxidant, lipase inhibitory, and α-amylase inhibitory activities in the extracts, respectively. Overall, this study revealed that underutilized protein sources such as WSPH, using the appropriate combination of enzymes, could result in the generation of new ingredients and compounds with powerful antioxidant and anti-obesity activities with promising applications as nutraceuticals or functional foods.Department of Agriculture Food and the Marinethe ERA-NET Cofund ERA HDHL | Ref. 69629

Neanderthal Fossils, Mobile Toolkit and a Hyena Den: The Archaeological Assemblage of Lateral Gallery 1 in Cova Del Gegant (NE Iberian Peninsula)

Lateral Gallery 1 (GL1) in Cova del Gegant is a Middle Palaeolithic assemblage yielding diagnostic Neanderthal remains, together with Mousterian tools and faunal remains. It is a good archive for evaluating the environmental conditions of the coastal areas during MIS 4 and MIS 3 in the NE of the Iberian Peninsula, and also the Neanderthals' behaviour and mobility. Here we provide a comprehensive assessment of all of the data available from GL1, such as lithics, human remains, fauna and chronostratigraphic details. The biotic ecofacts studied point to the development of a coastal plain in front of the cave and indicate that local conditions likely favoured a large variety of ecosystems characterised by open environments and woodland-edge taxa, and favoured repeated visits by humans during the Middle Palaeolithic. The evidence suggests that the gallery was mainly used by carnivores, such as hyenas, and also by Neanderthals as a brief stopping place, in view of the presence of transported and abandoned ergonomic lithic artifacts and/or the placement of bodies (or parts of bodies). The regional context suggests high human mobility and emphasises the variability of Neanderthal behaviour

Histochemistry evaluation of the oxidative stress and the antioxidant status in Cu-supplemented cattle

The aim of this paper is to evaluate at a histopathological level the effect of the most commonly used copper (Cu) supplementation (15 mg/kg dry matter (DM)) in the liver of intensively reared beef cattle. This was done by a histochemistry evaluation of (i) the antioxidant capacity in the liver – by the determination of metallothioneins (MT) and superoxide dismutase (SOD) expression – as well as (ii) the possible induction of oxidative damage – by the determination of inducible nitric oxide synthase (iNOS), nitrotyrosine (NITT), malondialdehyde (MDA) and 8-oxoguanine (8-oxo) – that (iii) could increase apoptotic cell death – determined by cytochrome-c (cyto-c), caspase 1 (casp1) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Liver samples from Cu-supplemented (15 mg Cu sulphate/kg DM, n = 5) and non-supplemented calves (n = 5) that form part of other experiments to evaluate Cu status were collected at slaughter and processed for immunohistochemistry and TUNEL. MT expression was diffuse and SOD showed slight changes although without statistical significance. iNOS and NITT positive (+) cells significantly increased, mainly around the central veins in the animals from the Cu-supplemented group, whereas no differences were appreciated for the rest of the oxidative stress and apoptosis markers. Under the conditions of this study, which are the conditions of the cattle raised in intensive systems in NW Spain and also many European countries, routinely Cu supplementation increased the risk of the animals to undergo subclinical Cu toxicity, with no significant changes in the Cu storage capacity and the antioxidant defensive system evaluated by MT and SOD expression, but with a significant and important increase of oxidative damage measured by iNOS and NITT. The results of this study indicated that iNOS and NITT could be used as early markers of initial pathological changes in the liver caused by Cu supplementation in cattle, although more studies in cattle under different levels of Cu supplementation are neededThis work was supported by the Xunta de Galicia (Spain) through contract 07MRU030261PR and by a research grant María Barbeito awarded to M. García-Vaquero by the Dirección Xeral de Investigación, Desenvolvemento e Innovación from Xunta de Galicia and to FSE (Fondo Social Europeo) funds through Programa de Recursos Humanos, do Plan Galego de Investigación, Desenvolvemento e Innovación Tecnolóxica de GaliciaS

Seasonal Variation of the Proximate Composition, Mineral Content, Fatty Acid Profiles and Other Phytochemical Constituents of Selected Brown Macroalgae

The main objective was to determine the chemical, phytochemical, fatty acid and mineral profiles of three commercially relevant brown macroalgae (Laminaria digitata, Laminaria hyperborea and Ascophyllum nodosum) collected each season for two years off the west coast of Ireland. All the chemical, phytochemical, fatty acid and minerals analysed varied significantly depending on the macroalgal species, season and year of collection. Overall, the protein contents of macroalgae were negatively correlated with carbohydrate content. Protein (2–11%) was at its highest during winter and/or spring, decreasing to a minimum during summer and/or autumn. The three macroalgal species analysed in this study had clearly differentiated fatty acid profiles. The concentration of fatty acids was higher in A. nodosum compared with both Laminaria species. The mineral profile of the three macroalgal species was rich in essential metals, particularly Ca, Mg and P, while the levels of I were approximately 9- to 10-fold higher in both Laminaria spp. compared with A. nodosum. The levels of toxic metals (Cd, Hg and Pb) in all the macroalgal species studied were low in the current study; while the levels of total As were high (49–64 mg/kg DW macroalgae) compared with previous reportsThis work was supported by Science Foundation Ireland (SFI) (grant number: 14/IA/2548)S

Integrin Linked Kinase (ILK) Downregulation as an Early Event During the Development of Metabolic Alterations in a Short-Term High Fat Diet Mice Model.

Background/Aims: Diabetes type 2, metabolic syndrome or non-alcoholic fatty liver disease are insulin resistance-related metabolic disorders, which lack a better prognosis before their full establishment. We studied the importance of the intracellular scaffold protein integrin linked kinaes (ILK) as a key modulator in the initial pathogenesis and the early progression of those insulin resistance- related disorders. Methods: Adult mice with a global transgenic downregulation of ILK expression (cKD-ILK) and littermates without that depletion (CT) were fed with either standard (STD) or high fat (HFD) diets during 2 and 6 weeks. Weights, blood glucose and other systemic biochemical parameters were determined in animals under fasting conditions and after glucose or pyruvate intraperitoneal injections to test their tolerance. In RNA or proteins extracted from insulin-sensitive tissues, we determined by reverse transcription?quantitative PCR and western blot the expression of ILK, metabolites transporters and other metabolism and inflammatory markers. Glucose uptake capacity was studied in freshly isolated tissues. Results: HFD feeding was able to early and progressively increase glycaemia, insulinemia, circulating glycerol, body weight gain, liver-mediated gluconeogenesis along this time lapse, but cKD-ILK have all these systemic misbalances exacerbated compared to CT in the same HFD time lapse. Interestingly, the tisular expression of ILK in HFD-fed CT was dramatically downregulated in white adipose tissue (WAT), skeletal muscle and liver at the same extent of the original ILK downregulation of cKD-ILK. We previously published that basal STD-fed cKD-ILK compared to basal STD-CT have different expression of glucose transporters GLUT4 in WAT and skeletal muscle. In the same STD-fed cKD-ILK, we observed here the increased expressions of hepatic GLUT2 and WAT pro-inflammatory cytokines TNF-? and MCP-1. The administration of HFD exacerbated the expression changes in cKD-ILK of these and other markers related to the imbalanced metabolism observed, such as WAT lipolysis (HSL), hepatic gluconeogenesis (PCK-1) and glycerol transport (AQP9). Conclusion: ILK expression may be taken as a predictive determinant of metabolic disorders establishment, because its downregulation seems to correlate with the early imbalance of glucose and glycerol transport and the subsequent loss of systemic homeostasis of these metabolites.Instituto de Salud Carlos III-ISCIIIComunidad de MadridFondo Europeo de Desarrollo Regional-FEDERInstituto Ramon y Cajal de Investigación Sanitária-IRYCISFundación Renal Iñigo Álvarez de Toledo-FRIA

The Integrin Beta1 Modulator Tirofiban Prevents Adipogenesis and Obesity by the Overexpression of Integrin-Linked Kinase: a Pre-Clinical Approach In Vitro and In Vivo

de Frutos, S., Griera, M., Hatem-Vaquero, M. et al. The integrin beta1 modulator Tirofiban prevents adipogenesis and obesity by the overexpression of integrin-linked kinase: a pre-clinical approach in vitro and in vivo. Cell Biosci 12, 10 (2022)Background: Obesity is caused by the enlargement of the white adipose tissue (WAT) depots, characterized by the hypertrophic enlargement of malfunctioning adipocytes within WAT which increases the storage of triglycerides (TG) in the lipid droplets (LD). Adipogenesis pathways as well as the expression and activity of some extracellular matrix receptors integrins are upregulated. Integrin?1 (INTB1) is the main isoform involved in WAT remodeling during obesity and insulin resistance-related diseases. We recently described Integrin Linked Kinase (ILK), a scafold protein recruited by INTB1, as an important mediator of WAT remodeling and insulin resistance. As the few approved drugs to fght obesity have brought long-term cardiovascular side efects and given that the consideration of INTB1 and/or ILK modulation as anti-obesogenic strategies remains unexplored, we aimed to evaluate the anti-obesogenic capacity of the clinically approved anticoagulant Tirofban (TF), stated in preclinical studies as a cardiovascular protector. Methods: Fully diferentiated adipocytes originating from C3H10T1/2 were exposed to TF and were co-treated with specifc INTB1 blockers or with siRNA-based knockdown ILK expression. Lipid-specifc dyes were used to determine the TG content in LD. The genetic expression pattern of ILK, pro-infammatory cytokines (MCP1, IL6), adipogenesis (PPAR?, Leptin), thermogenesis (UCP1), proliferation (PCNA), lipid metabolism (FASN, HSL, ATGL), and metabolite trans porters (FABP4, FAT, AQP7) were detected using quantitative PCR. Cytoskeletal actin polymerization was detected by confocal microscopy. Immunoblotting was performed to detect INTB1 phosphorylation at Thr788/9 and ILK activity as phosphorylation levels of protein kinase B (AKT) in Ser473 and glycogen synthase kinase 3? (GSK3?) at Ser9. TF was intraperitoneally administered once per day to wildtype and ILK knockdown mice (cKDILK) challenged with a high-fat diet (HFD) or control diet (STD) for 2 weeks. Body and WAT weight gains were compared. The expression of ILK and other markers was determined in the visceral epididymal (epi) and inguinal subcutaneous (sc) WAT. Results: TF reduced TG content and the expression of adipogenesis markers and transporters in adipocytes, while UCP-1 expression was increased and the expression of lipases, cytokines or PCNA was not afected. Mechanistically, TF rapidly increased and faded the intracellular phosphorylation of INTB1 but not AKT or GSK3?. F-actin levels were rapidly decreased, and INTB1 blockade avoided the TF efect. After 24 h, ILK expression and phosphorylation rates of AKT and GSK3? were upregulated, while ILK silencing increased TG content. INTB1 blockade and ILK silencing avoided TF efects on the TG content and the transcriptional expression of PPAR? and UCP1. In HFD-challenged mice, the systemic administration of TF for several days reduced the weight gain on WAT depots. TF reduced adipogenesis and pro-infammatory biomarkers and increased lipolysis markers HSL and FAT in epiWAT from HFD, while increased UCP1 in scWAT. In both WATs, TF upregulated ILK expression and activity, while no changes were observed in other tissues. In HFD-fed cKDILK, the blunted ILK in epiWAT worsened weight gain and avoided the anti-obesogenic efect of in vivo TF administration. Conclusions: ILK downregulation in WAT can be considered a biomarker of obesity establishment. Via an INTB1-ILK axis, TF restores malfunctioning hypertrophied WAT by changing the expression of adipocyte-related genes, increas ing ILK expression and activity, and reducing TG storage. TF prevents obesity, a property to be added to its anticoagu lant and cardiovascular protective advantages.Instituto de Salud Carlos IIIComunidad de MadridFondo Europeo de Desarrollo Regional-FEDE

Use of an NIR MEMS spectrophotometer and visible/NIR hyperspectral imaging systems to predict quality parameters of treated ground peppercorns

peer-reviewedThe aim of this study was to investigate the potential of a micro-electromechanical NIR spectrophotometer (NIR-MEMS) and visible (Vis)/NIR hyperspectral imaging (HSI) systems to predict the moisture content, antioxidant capacity (DPPH, FRAP) and total phenolic content (TPC) of treated ground peppercorns. Partial least squares (PLS) models were developed using spectra from peppercorns treated with hot-air, microwave and cold plasma. The spectra were acquired using three spectroscopy systems: NIR-MEMS (1350–1650 nm), Vis-NIR HSI (450–950 nm) and NIR HSI (957–1664 nm). Very good predictions of TPC (RPD > 3.6) were achieved using NIR-MEMS. The performance of models developed using Vis-NIR HSI and NIR HSI were good or very good for DPPH (RPD > 3.0), FRAP (RPD >2.9) and TPC (RPD > 3.8). This study demonstrated the potential of NIR-MEMS and Vis-NIR/NIR HSI to predict the moisture content, antioxidant capacity and total phenolic content of peppercorns. The spectroscopy technologies investigated are suitable for use as in-line PAT tools to facilitate improved process control and understanding during peppercorn processing

Consumer knowledge and attitudes towards microalgae as food: The case of Spain

Spanish consumers’ knowledge and attitudes towards microalgae as food were assessed using a specific questionnaire - a total of 3,084 Spanish consumers over 18 years old were considered in the current study. Overall, the questionnaire revealed that there is a huge lack of knowledge on basic aspects of microalgae, especially within the youngest respondents (18-24 years old) and for those aged over 65 years old. Approximately 85% of all respondents scored the statement “There is a big lack of information about microalgae” with either 4 or 5 (using a 5-point hedonic scale), demonstrating that microalgae and their potential applications are unknown to the majority of the Spanish population. Spanish consumers consider microalgae as: (i) sustainable and environmentally friendly, (ii) nutritious and healthy, and (iii) safe, which is of key importance for food products. The main reasons that were given for not consuming microalgae were lack of information available on the product and lack of consuming habit. The current study also demonstrated that increasing consumer’s knowledge on microalgae has the potential to increase consumer choice and market shares of microalgae-enriched products. Results reported herein could be extrapolated to other European countries, although perception and attitudes towards food can vary depending on cultural considerations

A Multicentric, Open-Label, Randomized, Comparative Clinical Trial of Two Different Doses of Expanded hBM-MSCs Plus Biomaterial versus Iliac Crest Autograft, for Bone Healing in Nonunions after Long Bone Fractures: Study Protocol

ORTHOUNION is a multicentre, open, comparative, three-arm, randomized clinical trial (EudraCT number 2015-000431-32) to compare the efficacy, at one and two years, of autologous human bone marrow-derived expanded mesenchymal stromal cell (hBM-MSC) treatments versus iliac crest autograft (ICA) to enhance bone healing in patients with diaphyseal and/or metaphysodiaphyseal fracture (femur, tibia, and humerus) status of atrophic or oligotrophic nonunion (more than 9 months after the acute fracture, including recalcitrant cases after failed treatments). The primary objective is to determine if the treatment with hBM-MSCs combined with biomaterial is superior to ICA in obtaining bone healing. If confirmed, a secondary objective is set to determine if the dose of 100 × 106 hBM-MSCs is noninferior to that of 200 × 106 hBM-MSCs. The participants (n = 108) will be randomly assigned to either the experimental low dose (n = 36), the experimental high dose (n = 36), or the comparator arm (n = 36) using a central randomization service. The trial will be conducted in 20 clinical centres in Spain, France, Germany, and Italy under the same clinical protocol. The confirmation of superiority for the proposed ATMP in nonunions may foster the future of bone regenerative medicine in this indication. On the contrary, absence of superiority may underline its limitations in clinical use