Bacillus coagulans GBI-30, 6086 (BC30) improves lactose digestion in rats exposed to a high-lactose meal.

PURPOSE: Bacillus coagulans GBI-30, 6086 (BC30) was previously shown to improve nutrient digestibility and amino acid absorption from milk protein in vitro. However, the effect of supplementation with this probiotic on lactose digestibility has not yet been evaluated in vivo. METHODS: Wistar female rats were exposed to an acute high-lactose diet (LD; 35% lactose) meal challenge after 7 days of administration of BC30 (LD-BC; n = 10) or vehicle (LD-C; n = 10). Rats treated with vehicle and exposed to control diet (CD; 35% corn starch) meal were used as controls (CD-C; n = 10). Carbohydrate oxidation (CH_OX) and lipid oxidation (L_OX) were monitored by indirect calorimetry before and after lactose challenge. After the challenge, rats were treated daily with vehicle or probiotic for an additional week and were fed with CD or LD ad libitum to determine the effects of BC30 administration in a lactose-induced diarrhoea and malnutrition model. RESULTS: LD-C rats showed lower CH_OX levels than CD rats, while LD-BC rats showed similar CH_OX levels compared to CD rats during the lactose challenge, suggesting a better digestion of lactose in the rats supplemented with BC30. BC30 completely reversed the increase in the small intestine length of LD-C animals. LD-BC rats displayed increased intestinal mRNA Muc2 expression. No significant changes were observed due to BC30 administration in other parameters, such as serum calprotectin, intestinal MPO activity, intestinal A1AT and SGLT1 levels or intestinal mRNA levels of Claudin2 and Occludin. CONCLUSION: Treatment with BC30 improved the digestibility of lactose in an acute lactose challenge and ameliorated some of the parameters associated with lactose-induced malnutrition.

A mixture of four dietary fibres ameliorates adiposity and improves metabolic profile and intestinal health in cafeteria-fed obese rats: an integrative multi-omics approach.

The aim of this study was to assess the effects of a mixture of four dietary fibers on obese rats. Four groups of male Wistar rats were fed with either standard chow (STD) or cafeteria diet (CAF) and were orally supplemented with either fibre mixture (2 g kg-1 of body weight) (STD+F or CAF+F groups) or vehicle (STD+VH or CAF+VH groups). We studied a wide number of biometric, biochemical, transcriptomic, metagenomic and metabolomic variables and applied an integrative multivariate approach based on multiple factor analysis and Pearson's correlation analysis. A significant reduction in body weight, adiposity, HbA1c and HDL-cholesterol serum levels, and colon MPO activity was observed, whereas cecal weight and small intestine length:weight ratio were significantly increased in F-treated groups compared to control animals. CAF+F rats displayed a significant enhancement in energy expenditure, fat oxidation and fresh stool weight, and a significant reduction in adiponectin and LPS serum levels, compared to control group. Animals in STD+F group showed reduced serum LDL-cholesterol levels and a significant reduction in total cholesterol levels in the liver compared to STF+VH group. The intervention effect was reflected at the metabolomic (i.e., production of short-chain fatty acids, phenolic acids, and amino acids), metagenomic (i.e., modulation of Ruminococcus and Lactobacillus genus) and transcriptomic (i.e., expression of tight junctions and proteolysis) levels. Altogether, our integrative multi-omics approach highlights the potential of supplementation with a mixture of fibers to ameliorate the impairments triggered by obesity in terms of adiposity, metabolic profile, and intestinal health.

Identification of novel antihypertensive peptides from wine lees hydrolysate.

Enzymatic-assisted extraction using Flavourzyme® has been demonstrated to be a useful methodology to obtain wine lees (WL) enriched in phenolic compounds and with enhanced antihypertensive activity. Nevertheless, taking into account that Flavourzyme® possess proteolytic activity, the release of bioactive peptides should not be ruled out. In this study, we investigate the presence of antihypertensive peptides in the WL hydrolysate. Peptides were separated into fractions by ultrafiltration and RP-HPLC. Next, peptide identification by nano-HPLC-(Orbitrap)MS/MS was performed in the fractions showing the highest angiotensin-converting enzyme inhibitory (ACEi) activities. Six peptides were identified; three of them showing ACEi (IC50) values lower than 20 µM. The peptide antihypertensive effect was evaluated in spontaneously hypertensive rats at an oral dose of 10 mg/kg bw. Peptides FKTTDQQTRTTVA, NPKLVTIV, TVTNPARIA, LDSPSEGRAPG and LDSPSEGRAPGAD exhibited antihypertensive activity, confirming that they could contribute to the blood pressure-lowering effect of the WL hydrolysate. These peptides have a great potential as functional ingredients to manage hypertension.

The Catalan Surveillance Network of SARS-CoV-2 in Sewage: design, implementation, and performance.

Wastewater-based epidemiology has shown to be an efficient tool to track the circulation of SARS-CoV-2 in communities assisted by wastewater treatment plants (WWTPs). The challenge comes when this approach is employed to help Health authorities in their decision-making. Here, we describe the roadmap for the design and deployment of SARSAIGUA, the Catalan Surveillance Network of SARS-CoV-2 in Sewage. The network monitors, weekly or biweekly, 56 WWTPs evenly distributed across the territory and serving 6 M inhabitants (80% of the Catalan population). Each week, samples from 45 WWTPs are collected, analyzed, results reported to Health authorities, and finally published within less than 72 h in an online dashboard ( https://sarsaigua.icra.cat ). After 20 months of monitoring (July 20-March 22), the standardized viral load (gene copies/day) in all the WWTPs monitored fairly matched the cumulative number of COVID-19 cases along the successive pandemic waves, showing a good fit with the diagnosed cases in the served municipalities (Spearman Rho = 0.69). Here we describe the roadmap of the design and deployment of SARSAIGUA while providing several open-access tools for the management and visualization of the surveillance data.

A multifunctional ingredient for the management of metabolic syndrome in cafeteria diet-fed rats.

The body weight-lowering properties of a multifunctional ingredient (MIX) based on conjugated linoleic acid at low doses, the flavonoids proanthocyanidins and anthocyanidins and the chicken feet hydrolysate Hpp11 have been previously reported. The aim of this study was to evaluate the effect of long-term administration of MIX on other cardiometabolic risk factors associated with metabolic syndrome (MetS) in rats fed a cafeteria diet (CAF). Male Wistar rats were fed CAF for 11 weeks, and during the last 3 weeks, animals were orally administered MIX or vehicle. Lipid tolerance tests were performed before and after MIX administration. At the end of the experimental period, serum and inguinal white adipose tissue (iWAT) metabolism were analyzed by metabolomics and biochemical approaches. The metabolite signature of serum and iWAT significantly changed after 3 weeks of MIX administration, suggesting an improvement in lipid and glucose homeostasis in these animals. In addition, MIX also exhibited significant antihypertensive properties. These results suggest that MIX could be a good candidate to ameliorate the cardiometabolic risk factors related to MetS.

Changes in arterial blood pressure caused by long-term administration of grape seed proanthocyanidins in rats with established hypertension.

The development of hypertension (HTN) in cafeteria (CAF) diet-fed rats was demonstrated to be attenuated after grape seed proanthocyanidin extract (GSPE) administration. However, the long-term antihypertensive effect of GSPE in animals with established HTN has not been investigated. Thus, the aim of this study was to evaluate if GSPE shows a blood pressure-lowering effect in hypertensive rats after its administration for 3 weeks. Wistar rats were fed a standard or CAF diet for 12 weeks, and during the last 3 weeks, animals were administered vehicle, captopril or a low dose of GSPE (25 mg per kg body weight, bw). Both systolic and diastolic blood pressure (SBP and DBP) were monitored weekly. The liver reduced glutathione (GSH) levels, plasma angiotensin converting enzyme activity and endothelial gene expression of eNOS, KLF-2, Sirt-1, NOX4 and ET-1 were studied at the end-point. The results demonstrated that 3 weeks of CAF diet administration with 25 mg per kg bw GSPE significantly reduced SBP and DBP in hypertensive rats. GSPE induced the upregulation of Sirt-1 gene expression and downregulated the vasoconstrictor ET-1, suggesting the vasoprotective effect of GSPE and increased the antioxidant GSH activity. The administration of 25 mg per kg bw GSPE for 3 weeks significantly reduced BP in CAF diet fed animals with established HTN.

Implication of Opioid Receptors in the Antihypertensive Effect of a Novel Chicken Foot-Derived Peptide.

The peptide AVFQHNCQE demonstrated to produce nitric oxide-mediated antihypertensive effect. This study investigates the bioavailability and the opioid-like activity of this peptide after its oral administration. For this purpose, in silico and in vitro approaches were used to study the peptide susceptibility to GI digestion. In addition, AVFQHNCQE absorption was studied both in vitro by using Caco-2 cell monolayers and in vivo evaluating peptide presence in plasma from Wistar rats by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and by ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Both in vivo and in vitro experiments demonstrated that peptide AVFQHNCQE was not absorbed. Thus, the potential involvement of opioid receptors in the BP-lowering effect of AVFQHNCQE was studied in the presence of opioid receptors-antagonist Naloxone. No changes in blood pressure were recorded in rats administered Naloxone, demonstrating that AVFQHNCQE antihypertensive effect is mediated through its interaction with opioid receptors. AVFQHNCQE opioid-like activity would clarify the antihypertensive properties of AVFQHNCQE despite its lack of absorption.

Effect of Hesperidin on Cardiovascular Disease Risk Factors: The Role of Intestinal Microbiota on Hesperidin Bioavailability.

Recently, hesperidin, a flavonone mainly present in citrus fruits, has emerged as a new potential therapeutic agent able to modulate several cardiovascular diseases (CVDs) risk factors. Animal and in vitro studies demonstrate beneficial effects of hesperidin and its derived compounds on CVD risk factors. Thus, hesperidin has shown glucose-lowering and anti-inflammatory properties in diabetic models, dyslipidemia-, atherosclerosis-, and obesity-preventing effects in CVDs and obese models, and antihypertensive and antioxidant effects in hypertensive models. However, there is still controversy about whether hesperidin could contribute to ameliorate glucose homeostasis, lipid profile, adiposity, and blood pressure in humans, as evidenced by several clinical trials reporting no effects of treatments with this flavanone or with orange juice on these cardiovascular parameters. In this review, we focus on hesperidin's beneficial effects on CVD risk factors, paying special attention to the high interindividual variability in response to hesperidin-based acute and chronic interventions, which can be partly attributed to differences in gut microbiota. Based on the current evidence, we suggest that some of hesperidin's contradictory effects in human trials are partly due to the interindividual hesperidin variability in its bioavailability, which in turn is highly dependent on the α-rhamnosidase activity and gut microbiota composition.

Novel Antihypertensive Peptides Derived from Chicken Foot Proteins.

SCOPE: Chicken foot proteins have recently been demonstrated by the group to be a great source of hydrolysates with antihypertensive properties. The aim of this study was to isolate and identify angiotensin I-converting enzyme inhibitory (ACEI) peptides from chicken foot hydrolysate Hpp11 and to test their antihypertensive properties. METHODS AND RESULTS: Peptides are separated into fractions according to their molecular size and hydrophobicity by ultrafiltration and RP-HPLC, respectively. Subsequent peptide identification in the two fractions that present the highest ACEI activities is carried out by HPLC-MS. Ten of the identified peptides are synthesized and five of them show ACEI (IC50 ) values lower than 100 µm. The antihypertensive effects of these ACEI peptides after oral administration is evaluated in spontaneously hypertensive rats. The peptides AVFQHNCQE and QVGPLIGRYCG exhibit antihypertensive activity when administered at an oral dose of 10 mg kg-1 body weight. The maximal decrease in systolic blood pressure is recorded 6 h after their administration (-25.07 ± 4.21 and -10.94 ± 1.96 mmHg, respectively). CONCLUSION: These results suggest that AVFQHNCQE and QVGPLIGRYCG could be used as functional ingredients with antihypertensive effects, although it would be necessary to perform bioavailability and clinical studies to demonstrate their efficiency in humans.

Optimization of extraction methods for characterization of phenolic compounds in apricot fruit (Prunus armeniaca).

Fruits are rich in phenolic compounds with health-promoting activities. However, phenolic profiles vary between fruits. Hence, specific extraction methods are required for accurate profiling of the functional compounds. This study aims to develop an optimised method by response surface methodology to extract phenolics from apricots (Prunus armeniaca) and correctly characterise apricots' phenolic profile. For this, the effects of the solid-to-liquid ratio, temperature, extraction solvent, extraction time and sequential extraction steps on the extraction of major phenolic families were investigated. Methanol- and ethanol-based extractions were suitable, although methanol was the optimal solvent. The optimised extraction conditions were 20 g mL-1, 38 °C and 72% methanol (1% formic acid). When this method was used in apricots, the characterisation of their phenolic profile by HPLC-ESI-MS/MS showed a higher extraction of phenolic compounds than other studies in the literature that use non-specific extraction methods. The developed method is fast and economically feasible for accurate characterisation of the phenolic profile of apricot fruits and thus can be routinely used to extract apricot phenolic compounds for their characterisation.

Optimization and characterization of Royal Dawn cherry (Prunus avium) phenolics extraction.

To correlate the beneficial effects of cherry consumption with their phenolic composition, a full and precise characterization is required. However, there is not a specific method to fully extract all phenolic compounds from sweet cherries. Thus, this study aimed to optimize the extraction of sweet cherry phenolics by response surface methodology and fully characterize the phenolic profile of Royal Dawn sweet cherries by HPLC-ESI-MS/MS. Extraction conditions were evaluated and optimized to 55 °C, MeOH 72%, 12 mL/g in two extraction steps. Royal Dawn sweet cherries presented rutin as the predominant phenolic compound, unlike most sweet cherry varieties. Additionally, ethanol was evaluated as a replacement solvent, obtaining lower extraction rates, especially for anthocyanins. However, in terms of total amounts, non-anthocyanin compounds were similarly extracted. The developed methodology was fast and can be routinely used in the evaluation of the phenolic profile of sweet cherries and to produce phenolic-rich extracts for the food industry.

A comparative study on the bioavailability of phenolic compounds from organic and nonorganic red grapes.

The health-promoting functions of fruit phenolic compounds are mainly attributed to their metabolites. The organic cultivation of fruits is becoming increasingly popular. Thus, this study evaluates whether the differences in red Grenache grapes derived from organic culture conditions influence the bioavailability and metabolism of phenolic compounds in rats. Organic and nonorganic (conventional) red Grenache grapes (OG and CG, respectively) were characterized and administered to Wistar rats (65 mg gallic acid equivalents/kg bw). Serum was recollected at different time points, and the phenolic metabolites were quantified by HPLC-ESI-MS/MS. The results showed that organic cultivation increased the oligomeric proanthocyanidin and anthocyanidin contents and decreased the content of free flavanols and dietary fiber. The serum profile of OG-administered rats showed higher metabolite concentrations at 2 h and reduced metabolite concentration at 24 h compared with the CG-administered rats. Thus, this particular serum kinetic behavior might influence the bioactivity of their phenolic compounds.

Exposure of Fischer 344 rats to distinct photoperiods influences the bioavailability of red grape polyphenols.

The consumption of grapes, rich in polyphenols, have several health effects. These effects are mainly attributed to the polyphenol metabolites generated after their ingestion. Several factors that affects host's physiology can modulate the bioavailability of grape polyphenols and, in turn, their effects. Mammals undergo physiological and metabolic changes due to the different day length (photoperiod) within a year. Thus, the aim of this study is to investigate if the bioavailability of phenolic compounds from the same red grapes with a different polyphenol profile (i.e. red grape produced organically (OG) or non-organically (conventional, CG)) differs with the photoperiod exposure. For this, the serum phenolic metabolites of Fischer 344 rats keep at different photoperiods (18, 12 or 6 h of light per day) and administered with OG or CG for 10-week were profiled by HPLC-MS/MS. Our results indicated that rats administered with OG reported a higher total serum metabolite concentration independent of the photoperiod exposure and CG-administered rats showed a more varied serum metabolite profile depending of the photoperiod exposure. Those rats exposed to 6 h of light per day, which emulates winter light conditions, presented a higher bioavailability of grape phenolics. Therefore, grape cultivar and animal photoperiod exposure condition grape phenolics' bioavailability.

Optimization of a polyphenol extraction method for sweet orange pulp (Citrus sinensis L.) to identify phenolic compounds consumed from sweet oranges.

The consumption of sweet oranges has been linked to several health benefits, many of which are attributed to hesperidin, a flavanone that is present in high amounts in these fruits. However, other phenolic compounds can contribute to the bioactivity of sweet orange. To link those effects to their phenolic profile, the complete characterization of the phenolic profile is mandatory. Although many studies have profiled the phenolic composition of orange juices, their pulps, which retain phenolic compounds, are overlooked. This fact is particularly relevant because dietary guidelines recommend the consumption of whole fruits. Therefore, this study aimed to develop a specific method for the optimal extraction of phenolics from orange pulp and to use this method to characterize these fruits grown at different locations by HPLC-ESI-MS/MS. The extraction conditions that reported the highest total polyphenol content (TPC) and hesperidin contents were 20 mL/g, 55 °C, and 90% methanol. The extraction time and number of sequential steps were further evaluated and optimized as 20 min and two extraction steps, respectively. Although lower extraction rates were achieved when using ethanol as the extraction solvent, high TPC and hesperidin yields were obtained, suggesting the potential use of this methodology to produce phenolic-rich extracts for the food industry. By applying the optimized methodology and analyzing the extracts by HPLC-ESI-MS/MS, geographic cultivation regions were demonstrated to affect the phenolic profiles of oranges. In short, we developed a quick, easy-to-perform methodology that can be used to extract orange phenolics from pulp for their identification and quantification and to evaluate the factors that affect the phenolic profile in sweet orange pulps.

Evidence that Nitric Oxide is Involved in the Blood Pressure Lowering Effect of the Peptide AVFQHNCQE in Spontaneously Hypertensive Rats.

AVFQHNCQE is an antihypertensive nonapeptide obtained from a chicken foot protein hydrolysate. The present study aims to investigate the mechanisms involved in its blood pressure (BP)-lowering effect. Male (17⁻20 weeks old) spontaneously hypertensive rats (SHR) were used in this study. Rats were divided into two groups and orally administered water or 10 mg/kg body weight (bw) AVFQHNCQE. One hour post-administration, animals of both groups were intra-peritoneally treated with 1 mL of saline or with 1 mL of saline containing 30 mg/kg bw Nω-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthesis, or with 1 mL of saline containing 5 mg/kg bw indomethacin, which is an inhibitor of prostacyclin synthesis (n = 6 per group). Systolic BP was recorded before oral administration and six hours after oral administration. In an additional experiment, SHR were administered water or 10 mg/kg bw AVFQHNCQE (n = 6 per group) and sacrificed six hours post-administration to study the mechanisms underlying the peptide anti-hypertensive effect. Moreover, the relaxation caused by AVFQHNCQE in isolated aortic rings from Sprague-Dawley rats was evaluated. The BP-lowering effect of the peptide was not changed after indomethacin administration but was completely abolished by L-NAME, which demonstrates that its anti-hypertensive effect is mediated by changes in endothelium-derived NO availability. In addition, AVFQHNCQE administration downregulated aortic gene expression of the vasoconstrictor factor endothelin-1 and the endothelial major free radical producer NADPH. Moreover, while no changes in plasma ACE activity were observed after its administration, liver GSH levels were higher in the peptide-treated group than in the water group, which demonstrates that AVFQHNCQE presents antioxidant properties.

Dose-Related Antihypertensive Properties and the Corresponding Mechanisms of a Chicken Foot Hydrolysate in Hypertensive Rats.

The antihypertensive properties of different doses of a chicken foot hydrolysate, Hpp11 and the mechanisms involved in this effect were investigated. Spontaneously hypertensive rats (SHR) were administered water, Captopril (50 mg/kg) or Hpp11 at different doses (25, 55 and 85 mg/kg), and the systolic blood pressure (SBP) was recorded. The SBP of normotensive Wistar-Kyoto (WKY) rats administered water or Hpp11 was also recorded. Additionally, plasmatic angiotensin-converting enzyme (ACE) activity was determined in the SHR administered Hpp11. Moreover, the relaxation caused by Hpp11 in isolated aortic rings from Sprague-Dawley rats was evaluated. Hpp11 exhibited antihypertensive activity at doses of 55 and 85 mg/kg, with maximum activity 6 h post-administration. At this time, no differences were found between these doses and Captopril. Initial SBP values of 55 and 85 mg/kg were recovered 24 or 8 h post-administration, respectively, 55 mg/kg being the most effective dose. At this dose, a reduction in the plasmatic ACE activity in the SHR was found. However, Hpp11 did not relax the aortic ring preparations. Therefore, ACE inhibition could be the mechanism underlying Hpp11 antihypertensive effect. Remarkably, Hpp11 did not modify SBP in WKY rats, showing that the decreased SBP effect is specific to the hypertensive state.

Optimized Extraction by Response Surface Methodology Used for the Characterization and Quantification of Phenolic Compounds in Whole Red Grapes (Vitis vinifera).

Scientific research has focused on the characterization of bioactive polyphenols from grape seeds and skins, and the pulp has often been overlooked. However, since the beneficial properties of grapes are associated with the consumption of whole fruit, a full extraction and posterior characterization of the phenolic compounds in whole grapes is required to identify the involved bioactive compounds. Such methodologies are not currently available for the whole edible parts of red grapes. This study aimed to determine the best polyphenol extraction conditions of whole red grapes, and apply the method to characterize and quantify the polyphenol composition of three different grapes. The optimized conditions were 80 mL/g, 65% methanol (1% formic acid), 72 °C, and 100 min under agitation of 500 rpm. Also, methanol and ethanol were compared as extraction solvents, and methanol achieved statistically higher extraction rates for anthocyanins. The results of this work suggest a higher quantification of phenolic compounds when red grapes are analyzed whole, including the seeds, pulp, and skin.