STIM1 translocation to the nucleus protects cells from DNA damage.

DNA damage represents a challenge for cells, as this damage must be eliminated to preserve cell viability and the transmission of genetic information. To reduce or eliminate unscheduled chemical modifications in genomic DNA, an extensive signaling network, known as the DNA damage response (DDR) pathway, ensures this repair. In this work, and by means of a proteomic analysis aimed at studying the STIM1 protein interactome, we have found that STIM1 is closely related to the protection from endogenous DNA damage, replicative stress, as well as to the response to interstrand crosslinks (ICLs). Here we show that STIM1 has a nuclear localization signal that mediates its translocation to the nucleus, and that this translocation and the association of STIM1 to chromatin increases in response to mitomycin-C (MMC), an ICL-inducing agent. Consequently, STIM1-deficient cell lines show higher levels of basal DNA damage, replicative stress, and increased sensitivity to MMC. We show that STIM1 normalizes FANCD2 protein levels in the nucleus, which explains the increased sensitivity of STIM1-KO cells to MMC. This study not only unveils a previously unknown nuclear function for the endoplasmic reticulum protein STIM1 but also expands our understanding of the genes involved in DNA repair.

Evaluation of Parameters Which Influence Voluntary Ingestion of Supplements in Rats.

Drug safety and efficacy studies frequently use oral gavage, but repetitive usage may cause problems. Administration through voluntary ingestion represents an opportunity for refinement. We aimed to develop a protocol for voluntary ingestion of gelatin-based supplements in rats, assessing the influence of age, sex, fasting (4 h), and additives (vanilla, VF; sucralose, S), and to test it in lactating dams. Three-week-old and 5-month-old Sprague-Dawley rats were placed individually in an empty cage containing a gelatin cube and trained daily (5 days/week), recording the day the whole cube was consumed (latency). Rats trained prior to gestation were offered a gelatin containing 250 mg/kg cocoa shell extract (CSE) during lactation. Rats that did not eat the cube after 8 training days were considered non-habituated, with a proportion similar in young males (7.1%), young females (11.1%), and adult females (10.3%), but significantly higher in adult males (39.3%). Excluding non-habituated rats, latency was 2-3 days, without differences between young and adult rats (p = 0.657) or between males and females (p = 0.189). VF or VF + S in the gelatin did not modify latency, while fasting significantly reduced it in females (p = 0.007) but not in males (p = 0.501). During lactation, trained females ate the CSE-gelatin within 1-5 min without litter problems. Conclusions: Acceptance of a gelatin-based supplement is negatively influenced by male sex, facilitated by fasting, and not modified by additives. Training is remembered after 2 months and does not interfere with lactation. Gelatin-based voluntary ingestion is suitable to administer drugs that need to pass through the digestive system, ensuring adequate dosage, and is important to detect non-habituated rats prior to the study. The current protocol may be implemented by training the rats in their own cage.

Exploring the potential of phenolic compounds from the coffee pulp in preventing cellular oxidative stress after in vitro digestion.

The coffee pulp, a by-product of the coffee industry, contains a high concentration of phenolic compounds and caffeine. Simulated gastrointestinal digestion may influence these active compounds' bioaccessibility, bioavailability, and bioactivity. Understanding the impact of the digestive metabolism on the coffee pulp's phenolic composition and its effect on cellular oxidative stress biomarkers is essential. In this study, we evaluated the influence of in vitro gastrointestinal digestion of the coffee pulp flour (CPF) and extract (CPE) on their phenolic profile, radical scavenging capacity, cellular antioxidant activity, and cytoprotective properties in intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CPF and the CPE contained a high amount of caffeine and phenolic compounds, predominantly phenolic acids (3',4'-dihydroxycinnamoylquinic and 3,4-dihydroxybenzoic acids) and flavonoids (3,3',4',5,7-pentahydroxyflavone derivatives). Simulated digestion resulted in increased antioxidant capacity, and both the CPF and the CPE demonstrated free radical scavenging abilities even after in vitro digestion. The CPF and the CPE did not induce cytotoxicity in intestinal and hepatic cells, and both matrices exhibited the ability to scavenge intracellular reactive oxygen species. The coffee pulp treatments prevented the decrease of glutathione, thiol groups, and superoxide dismutase and catalase enzymatic activities evoked by tert-butyl hydroperoxide elicitation in IEC-6 and HepG2 cells. Our findings suggest that the coffee pulp could be used as a potent food ingredient for preventing cellular oxidative stress due to its high content of antioxidant compounds.

Radical Scavenging and Cellular Antioxidant Activity of the Cocoa Shell Phenolic Compounds after Simulated Digestion.

The cocoa industry generates a considerable quantity of cocoa shell, a by-product with high levels of methylxanthines and phenolic compounds. Nevertheless, the digestion process can extensively modify these compounds' bioaccessibility, bioavailability, and bioactivity as a consequence of their transformation. Hence, this work's objective was to assess the influence of simulated gastrointestinal digestion on the concentration of phenolic compounds found in the cocoa shell flour (CSF) and the cocoa shell extract (CSE), as well as to investigate their radical scavenging capacity and antioxidant activity in both intestinal epithelial (IEC-6) and hepatic (HepG2) cells. The CSF and the CSE exhibited a high amount of methylxanthines (theobromine and caffeine) and phenolic compounds, mainly gallic acid and (+)-catechin, which persisted through the course of the simulated digestion. Gastrointestinal digestion increased the antioxidant capacity of the CSF and the CSE, which also displayed free radical scavenging capacity during the simulated digestion. Neither the CSF nor the CSE exhibited cytotoxicity in intestinal epithelial (IEC-6) or hepatic (HepG2) cells. Moreover, they effectively counteracted oxidative stress triggered by tert-butyl hydroperoxide (t-BHP) while preventing the decline of glutathione, thiol groups, superoxide dismutase, and catalase activities in both cell lines. Our study suggests that the cocoa shell may serve as a functional food ingredient for promoting health, owing to its rich concentration of antioxidant compounds that could support combating the cellular oxidative stress associated with chronic disease development.

Changes in the cocoa shell dietary fiber and phenolic compounds after extrusion determine its functional and physiological properties.

The influence of different extrusion conditions on the cocoa shell (CS) dietary fiber, phenolic compounds, and antioxidant and functional properties was evaluated. Extrusion produced losses in the CS dietary fiber (3-26%), especially in the insoluble fraction, being more accentuated at higher temperatures (160 °C) and lower moisture feed (15-20%). The soluble fiber fraction significantly increased at 135 °C because of the solubilization of galactose- and glucose-containing insoluble polysaccharides. The extruded CS treated at 160 °C-25% of feed moisture showed the highest increase of total (27%) and free (58%) phenolic compounds, accompanied by an increase of indirect (10%) and direct (77%) antioxidant capacity. However, more promising results relative to the phenolic compounds' bioaccessibility after in vitro simulated digestion were observed for 135°C-15% of feed moisture extrusion conditions. The CS' physicochemical and techno-functional properties were affected by extrusion, producing extrudates with higher bulk density, a diminished capacity to hold oil (22-28%) and water (18-65%), and improved swelling properties (14-35%). The extruded CS exhibited increased glucose adsorption capacity (up to 2.1-fold, at 135 °C-15% of feed moisture) and α-amylase in vitro inhibitory capacity (29-54%), accompanied by an increase in their glucose diffusion delaying ability (73-91%) and their starch digestion retardation capacity (up to 2.8-fold, at 135 °C-15% of feed moisture). Moreover, the extruded CS preserved its cholesterol and bile salts binding capacity and pancreatic lipase inhibitory properties. These findings generated knowledge of the CS valorization through extrusion to produce foods rich in dietary fiber with improved health-promoting properties due to the extrusion-triggered fiber solubilization.

Valorization of coffee pulp as bioactive food ingredient by sustainable extraction methodologies.

Coffee pulp is an underutilized by-product of coffee industrial production rich in bioactive compounds such as phenolic compounds, caffeine, and dietary fiber. The widely known antioxidant, anti-inflammatory, anti-aging, antimicrobial and hepatoprotective health-promoting properties attributed to mentioned compounds enhance the use of coffee pulp as a bioactive food ingredient. Furthermore, the application of green sustainable extraction techniques pursuing highly efficient and selective extraction processes promotes this by-product exploitation in food science. Hence, this review gathers the available information relative to the impact of the extraction processes on the bioactive compound's recovery from coffee pulp, providing an overview of the most recent advances. An in-depth comparison workout between conventional and alternative extraction methods was performed to identify the most suitable techniques for coffee pulp valorization as functional ingredient until date. A critical discussion focused on advantages and drawbacks of the extraction methods applied to coffee pulp was included together a prospective of emerging extraction techniques.

Improvement of the Seminal Characteristics in Rams Using Agri-Food By-Products Rich in Phytomelatonin.

The aim of this study was to evaluate the effect of a phytomelatonin-rich diet, including by-products from the food industry, on ram sperm quality and seminal plasma composition. Melatonin content in several by-products before and after in vitro ruminal and abomasal digestion was determined by HPLC-ESI-MS/MS. Finally, 20% of a mix of grape pulp with pomegranate and tomato pomaces was included in the rams' diet, constituting the phytomelatonin-rich diet. Feeding the rams with this diet resulted in an increase in seminal plasma melatonin levels compared with the control group (commercial diet) in the third month of the study. In addition, percentages higher than those in the control group of morphologically normal viable spermatozoa with a low content of reactive oxygen species were observed from the second month onwards. However, the antioxidant effect does not seem to be exerted through the modulation of the antioxidant enzymes since the analysis of the activities of catalase, glutathione reductase and glutathione peroxidase in seminal plasma revealed no significant differences between the two experimental groups. In conclusion, this study reveals, for the first time, that a phytomelatonin-rich diet can improve seminal characteristics in rams.

Generation of mesenchymal stromal cells from urine-derived iPSCs of pediatric brain tumor patients.

Human induced pluripotent stem cells (iPSCs) provide a virtually inexhaustible source of starting material for next generation cell therapies, offering new opportunities for regenerative medicine. Among different cell sources for the generation of iPSCs, urine cells are clinically relevant since these cells can be repeatedly obtained by non-invasive methods from patients of any age and health condition. These attributes encourage patients to participate in preclinical and clinical research. In particular, the use of urine-derived iPSC products is a convenient strategy for children with brain tumors, which are medically fragile patients. Here, we investigate the feasibility of using urine samples as a source of somatic cells to generate iPSC lines from pediatric patients with brain tumors (BT-iPSC). Urinary epithelial cells were isolated and reprogrammed using non-integrative Sendai virus vectors harboring the Yamanaka factors KLF4, OCT3/4, SOX2 and C-MYC. After reprogramming, BT-iPSC lines were subject to quality assessment and were compared to iPSCs obtained from urine samples of non-tumor pediatric patients (nonT-iPSC). We demonstrated that iPSCs can be successfully derived from a small volume of urine obtained from pediatric patients. Importantly, we showed that BT-iPSCs are equivalent to nonT-iPSCs in terms of morphology, pluripotency, and differentiation capacity into the three germ layers. In addition, both BT-iPSCs and nonT-iPSCs efficiently differentiated into functional mesenchymal stem/stromal cells (iMSC) with immunomodulatory properties. Therefore, this study provides an attractive approach to non-invasively generate personalized iMSC products intended for the treatment of children with brain tumors.

Gastrointestinal fate of phenolic compounds and amino derivatives from the cocoa shell: An in vitro and in silico approach.

The objective of this study was to assess how in vitro gastrointestinal digestion influenced the bioaccessibility and potential bioavailability of phenolic compounds and methylxanthines in thecocoa shell (CS) in the form of flour (CSF) and aqueous extract (CSE). To comprehend how these phytochemicals behaved during gastrointestinal digestion, we also modeled in silico the colonic microbial biotransformation of the phenolic compounds in the CS. Different groups of phenolic compounds (mainly gallic andprotocatechuic acids, and catechin) and methylxanthines (theobromine and caffeine)could be found in the CS. Methylxanthines and phenolic compounds were released differently during gastrointestinal digestion. Whereas digestion triggered the release of hydroxybenzoic acids (67-73%) and flavan-3-ols (73-88%) during the intestinal phase, it also caused the degradation of flavonols and flavones. Besides, the release of phytochemicals was significantly influenced by the CS matrix type. Phenolic compounds were protected by the CSF matrix. Phenolic acids from CSF were more bioaccessible in the intestinal (1.2-fold, p < 0.05) and colonic (1.3-fold, p < 0.05) phases than those from the CSE. Methylxanthines were also more bioaccessible in the intestinal (1.8-fold, p < 0.01) and colonic phases (1.3-fold, p < 0.001) and bioavailable (1.8-fold, p < 0.001) in the CSF. Colonic metabolism demonstrated that the gut microbiota could biotransform non-absorbed phenolic compounds into other lower molecular weight and more bioavailable metabolites. These findings support the CS's potential as a source of bioaccessible, bioavailable, and active phytochemicals.

Internalized Amyloid-ß (1-42) Peptide Inhibits the Store-Operated Calcium Entry in HT-22 Cells.

Dysregulation in calcium signaling pathways plays a major role in the initiation of Alzheimer's disease (AD) pathogenesis. Accumulative experimental evidence obtained with cellular and animal models, as well as with AD brain samples, points out the high cytotoxicity of soluble small oligomeric forms of amyloid-ß peptides (Aß) in AD. In recent works, we have proposed that Aß-calmodulin (CaM) complexation may play a major role in neuronal Ca2+ signaling, mediated by CaM-binding proteins (CaMBPs). STIM1, a recognized CaMBP, plays a key role in store-operated calcium entry (SOCE), and it has been shown that the SOCE function is diminished in AD, resulting in the instability of dendric spines and enhanced amyloidogenesis. In this work, we show that 2 and 5 h of incubation with 2 µM Aß(1-42) oligomers of the immortalized mouse hippocampal cell line HT-22 leads to the internalization of 62 ± 11 nM and 135 ± 15 nM of Aß(1-42), respectively. Internalized Aß(1-42) oligomers colocalize with the endoplasmic reticulum (ER) and co-immunoprecipitated with STIM1, unveiling that this protein is a novel target of Aß. Fluorescence resonance energy transfer measurements between STIM1 tagged with a green fluorescent protein (GFP) and Aß(1-42)-HiLyte™-Fluor555 show that STIM1 can bind nanomolar concentrations of Aß(1-42) oligomers at a site located close to the CaM-binding site in STIM1. Internalized Aß(1-42) produced dysregulation of the SOCE in the HT-22 cells before a sustained alteration of cytosolic Ca2+ homeostasis can be detected, and is elicited by only 2 h of incubation with 2 µM Aß(1-42) oligomers. We conclude that Aß(1-42)-induced SOCE dysregulation in HT-22 cells is caused by the inhibitory modulation of STIM1, and the partial activation of ER Ca2+-leak channels.

Understanding the Gastrointestinal Behavior of the Coffee Pulp Phenolic Compounds under Simulated Conditions.

Numerous residues, such as the coffee pulp, are generated throughout coffee processing. This by-product is a source of antioxidant phytochemicals, including phenolic compounds and caffeine. However, the antioxidant properties of the phenolic compounds from the coffee pulp are physiologically limited to their bioaccessibility, bioavailability, and biotransformation occurring during gastrointestinal digestion. Hence, this study explored the phenolic and caffeine profile in the coffee pulp flour (CPF) and extract (CPE), their intestinal bioaccessibility through in vitro digestion, and their potential bioavailability and colonic metabolism using in silico models. The CPE exhibited a higher concentration of phenolic compounds than the CPF, mainly phenolic acids (protocatechuic, chlorogenic, and gallic acids), followed by flavonoids, particularly quercetin derivatives. Caffeine was found in higher concentrations than phenolic compounds. The antioxidant capacity was increased throughout the digestive process. The coffee pulp matrix influenced phytochemicals' behavior during gastrointestinal digestion. Whereas individual phenolic compounds generally decreased during digestion, caffeine remained stable. Then, phenolic acids and caffeine were highly bioaccessible, while flavonoids were mainly degraded. As a result, caffeine and protocatechuic acid were the main compounds absorbed in the intestine after digestion. Non-absorbed phenolic compounds might undergo colonic biotransformation yielding small and potentially more adsorbable phenolic metabolites. These results contribute to establishing the coffee pulp as an antioxidant food ingredient since it contains bioaccessible and potentially bioavailable phytochemicals with potential health-promoting properties.

Heterogeneity of In Vitro Expanded Mesenchymal Stromal Cells and Strategies to Improve Their Therapeutic Actions.

Beneficial properties of mesenchymal stromal cells (MSCs) have prompted their use in preclinical and clinical research. Accumulating evidence has been provided for the therapeutic effects of MSCs in several pathologies, including neurodegenerative diseases, myocardial infarction, skin problems, liver disorders and cancer, among others. Although MSCs are found in multiple tissues, the number of MSCs is low, making in vitro expansion a required step before MSC application. However, culture-expanded MSCs exhibit notable differences in terms of cell morphology, physiology and function, which decisively contribute to MSC heterogeneity. The changes induced in MSCs during in vitro expansion may account for the variability in the results obtained in different MSC-based therapy studies, including those using MSCs as living drug delivery systems. This review dissects the different changes that occur in culture-expanded MSCs and how these modifications alter their therapeutic properties after transplantation. Furthermore, we discuss the current strategies developed to improve the beneficial effects of MSCs for successful clinical implementation, as well as potential therapeutic alternatives.

Activating Effects of the Bioactive Compounds From Coffee By-Products on FGF21 Signaling Modulate Hepatic Mitochondrial Bioenergetics and Energy Metabolism in vitro.

Coffee by-products contain bioactive compounds that have been shown to have the capacity to modulate human metabolism. The goal of this study was to investigate the effects of the main bioactive compounds in coffee by-products and two aqueous extracts from the coffee husk and silverskin on the activation of fibroblast growth factor 21 (FGF21) signaling and the subsequent regulation of mitochondrial bioenergetics and lipid and glucose metabolism. HepG2 cells treated with palmitic acid (PA) were used in a non-alcoholic fatty liver disease (NAFLD) cell model. The bioactive compounds from coffee by-products (50 µmol L-1) and the aqueous extracts from the coffee silverskin and coffee husk (100 µg mL-1) increased ERK1/2 phosphorylation and the secretion of FGF21 (1.3 to 1.9-fold). Coffee by-products' bioactive compounds counteracted inflammation and PA-triggered lipotoxicity. Oxidative stress markers (ROS, mitochondrial superoxide, and NADPH oxidase) and the activity of antioxidant enzymes (superoxide dismutase and catalase) were modulated through the activation of Nrf2 signaling. Mitochondrial bioenergetics were regulated by enhancing respiration and ATP production via PGC-1α, and the expression of oxidative phosphorylation complexes increased. Coffee by-products' bioactive compounds decreased lipid accumulation (23-41%) and fatty acid synthase activity (32-65%) and triggered carnitine palmitoyltransferase-1 activity (1.3 to 1.7-fold) by activating AMPK and SREBP-1c pathways. The GLUT2 expression and glucose uptake were increased (58-111%), followed by a promoted glucokinase activity (55-122%), while glucose production and phosphoenolpyruvate carboxykinase activity were reduced due to IRS-1/Akt1 regulation. The bioactive compounds from coffee by-products, primarily chlorogenic and protocatechuic acids, could regulate hepatic mitochondrial function and lipid and glucose metabolism by activating FGF21 and related signaling cascades.

Phytochemicals from the Cocoa Shell Modulate Mitochondrial Function, Lipid and Glucose Metabolism in Hepatocytes via Activation of FGF21/ERK, AKT, and mTOR Pathways.

The cocoa shell is a by-product that may be revalorized as a source of bioactive compounds to prevent chronic cardiometabolic diseases. This study aimed to investigate the phytochemicals from the cocoa shell as targeted compounds for activating fibroblast growth factor 21 (FGF21) signaling and regulating non-alcoholic fatty liver disease (NAFLD)-related biomarkers linked to oxidative stress, mitochondrial function, and metabolism in hepatocytes. HepG2 cells treated with palmitic acid (PA, 500 µmol L-1) were used in an NAFLD cell model. Phytochemicals from the cocoa shell (50 µmol L-1) and an aqueous extract (CAE, 100 µg mL-1) enhanced ERK1/2 phosphorylation (1.7- to 3.3-fold) and FGF21 release (1.4- to 3.4-fold) via PPARα activation. Oxidative stress markers were reduced though Nrf-2 regulation. Mitochondrial function (mitochondrial respiration and ATP production) was protected by the PGC-1α pathway modulation. Cocoa shell phytochemicals reduced lipid accumulation (53-115%) and fatty acid synthase activity (59-93%) and prompted CPT-1 activity. Glucose uptake and glucokinase activity were enhanced, whereas glucose production and phosphoenolpyruvate carboxykinase activity were diminished. The increase in the phosphorylation of the insulin receptor, AKT, AMPKα, mTOR, and ERK1/2 conduced to the regulation of hepatic mitochondrial function and energy metabolism. For the first time, the cocoa shell phytochemicals are proved to modulate FGF21 signaling. Results demonstrate the in vitro preventive effect of the phytochemicals from the cocoa shell on NAFLD.

Investigating edible insects as a sustainable food source: nutritional value and techno-functional and physiological properties.

This work is aimed to evaluate the nutritional composition, and the techno-functional and in vitro physiological properties of flours made using six different insect species and the sensorial feasibility of including them in bakery products. The insect flours exhibited high protein and fat contents as their main components, highlighting the presence of chitin in ant samples. The techno-functional properties showed high oil holding, swelling, and emulsifying capacities in all the analysed insect flours, whereas their bulk density, hydration properties, and foaming capacity showed average values and no gelation capacity. Moreover, these edible insect flours exhibited effective hyperglycaemia and hyperlipidaemia properties, which together with their high antioxidant capacity are associated with beneficial in vitro physiological effects. The beetle and caterpillar flours stand out in these properties, and thus were selected to make a cupcake. The sensory evaluation confirmed that the edible beetle powder can be successfully included in baked goods to provide excellent sensory properties and very high acceptance. Thus, these insect flours may be of great interest to the food industry as a healthy source of protein, exerting a positive impact on functional and sensory food properties, and with a potential role in the prevention of diseases associated with hyperglycaemia and hyperlipidaemia.

Preclinical Safety Evaluation of Intranasally Delivered Human Mesenchymal Stem Cells in Juvenile Mice.

Mesenchymal stem cell (MSC)-based therapy is a promising therapeutic approach in the management of several pathologies, including central nervous system diseases. Previously, we demonstrated the therapeutic potential of human adipose-derived MSCs for neurological sequelae of oncological radiotherapy using the intranasal route as a non-invasive delivery method. However, a comprehensive investigation of the safety of intranasal MSC treatment should be performed before clinical applications. Here, we cultured human MSCs in compliance with quality control standards and administrated repeated doses of cells into the nostrils of juvenile immunodeficient mice, mimicking the design of a subsequent clinical trial. Short- and long-term effects of cell administration were evaluated by in vivo and ex vivo studies. No serious adverse events were reported on mouse welfare, behavioral performances, and blood plasma analysis. Magnetic resonance study and histological analysis did not reveal tumor formation or other abnormalities in the examined organs of mice receiving MSCs. Biodistribution study reveals a progressive disappearance of transplanted cells that was further supported by an absent expression of human GAPDH gene in the major organs of transplanted mice. Our data indicate that the intranasal application of MSCs is a safe, simple and non-invasive strategy and encourage its use in future clinical trials.

Revalorization of Coffee Husk: Modeling and Optimizing the Green Sustainable Extraction of Phenolic Compounds.

This study aimed to model and optimize a green sustainable extraction method of phenolic compounds from the coffee husk. Response surface methodology (RSM) and artificial neural networks (ANNs) were used to model the impact of extraction variables (temperature, time, acidity, and solid-to-liquid ratio) on the recovery of phenolic compounds. All responses were fitted to the RSM and ANN model, which revealed high estimation capabilities. The main factors affecting phenolic extraction were temperature, followed by solid-to-liquid ratio, and acidity. The optimal extraction conditions were 100 °C, 90 min, 0% citric acid, and 0.02 g coffee husk mL-1. Under these conditions, experimental values for total phenolic compounds, flavonoids, flavanols, proanthocyanidins, phenolic acids, o-diphenols, and in vitro antioxidant capacity matched with predicted ones, therefore, validating the model. The presence of chlorogenic, protocatechuic, caffeic, and gallic acids and kaemferol-3-O-galactoside was confirmed by UPLC-ESI-MS/MS. The phenolic aqueous extracts from the coffee husk could be used as sustainable food ingredients and nutraceutical products.

Extruded coffee parchment shows enhanced antioxidant, hypoglycaemic, and hypolipidemic properties by releasing phenolic compounds from the fibre matrix.

The dietary fibre and phenolic contents and the functional properties of extruded coffee parchment flour were studied to evaluate its possible use as an ingredient rich in dietary fibre (DF) with potential antioxidant, hypoglycaemic and hypolipidemic properties in extruded products. Coffee parchment flour treated at 160-175 °C and 25% moisture feed showed higher DF (84.3%) and phenolic contents (6.5 mg GAE per g) and antioxidant capacity (32.2 mg TE per g). The extrusion process favoured the release of phenolic compounds from the fibre matrix. Phytochemicals liberated during in vitro simulated digestion exhibited enhanced antioxidant capacity and attenuated reactive oxygen species in intestinal cells (IEC-6). However, the physicochemical and techno-functional properties were just affected by extrusion at high temperature, although extruded coffee parchment flours exhibited lower bulk density and higher swelling capacity than non-extruded ones. Extruded coffee parchment preserved the glucose adsorption capacity and enhanced the α-amylase in vitro inhibitory capacity (up to 81%). Moreover, extruded coffee parchment maintained the ability to delay glucose diffusion and exhibited improved capacity to retard starch digestion in the gastrointestinal tract. The extrusion of coffee parchment flours preserved the cholesterol-binding ability and augmented the capacity of this ingredient to bind bile salts, favouring the inhibition of pancreatic lipase by coffee parchment. These discoveries generate knowledge of the valorisation of coffee parchment as a food dietary fibre ingredient with antioxidant, hypoglycaemic, and hypolipidemic properties that are enhanced by the release of phenolic compounds from the fibre matrix through the production of extruded products.

Phytochemicals: Dietary Sources, Innovative Extraction, and Health Benefits.

Plants are the main natural source of numerous phytochemicals, although only a certain amount have been isolated and identified [...].

STIM1 Deficiency Leads to Specific Down-Regulation of ITPR3 in SH-SY5Y Cells.

STIM1 is an endoplasmic reticulum (ER) protein that modulates the activity of a number of Ca2+ transport systems. By direct physical interaction with ORAI1, a plasma membrane Ca2+ channel, STIM1 activates the ICRAC current, whereas the binding with the voltage-operated Ca2+ channel CaV1.2 inhibits the current through this latter channel. In this way, STIM1 is a key regulator of Ca2+ signaling in excitable and non-excitable cells, and altered STIM1 levels have been reported to underlie several pathologies, including immunodeficiency, neurodegenerative diseases, and cancer. In both sporadic and familial Alzheimer's disease, a decrease of STIM1 protein levels accounts for the alteration of Ca2+ handling that compromises neuronal cell viability. Using SH-SY5Y cells edited by CRISPR/Cas9 to knockout STIM1 gene expression, this work evaluated the molecular mechanisms underlying the cell death triggered by the deficiency of STIM1, demonstrating that STIM1 is a positive regulator of ITPR3 gene expression. ITPR3 (or IP3R3) is a Ca2+ channel enriched at ER-mitochondria contact sites where it provides Ca2+ for transport into the mitochondria. Thus, STIM1 deficiency leads to a strong reduction of ITPR3 transcript and ITPR3 protein levels, a consequent decrease of the mitochondria free Ca2+ concentration ([Ca2+]mit), reduction of mitochondrial oxygen consumption rate, and decrease in ATP synthesis rate. All these values were normalized by ectopic expression of ITPR3 in STIM1-KO cells, providing strong evidence for a new mode of regulation of [Ca2+]mit mediated by the STIM1-ITPR3 axis.