Development of an Innovative Berberine Food-Grade Formulation with an Ameliorated Absorption: In Vitro Evidence Confirmed by Healthy Human Volunteers Pharmacokinetic Study

Objective. To evaluate in vitro solubility, bioaccessibility, and cytotoxic profile, together with a pharmacokinetic profile by oral administration to healthy volunteers of a novel food-grade berberine formulation (BBR-PP, i.e., berberine Phytosome®). Results. An in vitro increase of solubility in simulated gastric and intestinal fluids and an improved bioaccessibility at intestinal level along with a lower cytotoxicity with respect to berberine were observed with BBR-PP. The pharmacokinetic profile of the oral administration to healthy volunteers confirmed that berberine Phytosome® significantly ameliorated berberine absorption, in comparison to unformulated berberine, without any observed side effects. The berberine plasma concentrations observed with both doses of BBR-PP were significantly higher than those seen after unformulated berberine administration, starting from 45 min (free berberine) and 30 min (total berberine). Furthermore, BBR-PP improved berberine bioavailability (AUC) was significantly higher, around 10 times on molar basis and with observed dose linearity, compared to the unformulated berberine. Conclusion. These findings open new perspectives on the use of this healthy berberine formulation in metabolic discomforts.

Microbiological Quality and Resistance to an Artificial Gut Environment of Two Probiotic Formulations

The quality control of probiotic products is the focus of numerous organizations worldwide. Several studies have highlighted the poor microbiological quality of many commercial probiotic formulations in terms of the identity of the contained microorganisms, viability, and purity, thus precluding the expected health benefits and representing a potential health risk for consumers. In this paper, we analyzed the contents of two probiotic formulations, one composed of an encapsulated mixture of lactobacilli and bifidobacteria, and one by a lyophilized yeast. The microorganisms contained in the products were quantified and identified using up-to-date methodologies, such as MALDI-TOF MS and metagenomic analysis. Moreover, as acid and bile tolerance is included among the criteria used to select probiotic microorganisms, in vitro tests were performed to evaluate the behavior of the formulations in conditions mimicking the harsh gastric environment and the intestinal fluids. Our results indicate the high quality of the formulations in terms of the enumeration and identification of the contained organisms, as well as the absence of contaminants. Moreover, both products tolerated the acidic conditions well, with encapsulation providing further protection for the microorganisms. A good tolerance to the simulated artificial intestinal conditions was also evidenced for both preparations.

Adsorption and Release Characteristics of Purified and Non-Purified Clinoptilolite Tuffs towards Health-Relevant Heavy Metals

The occurrence of health-relevant contaminants in water has become a severe global problem. For treating heavy-metal-polluted water, the use of zeolite materials has been extended over the last decades, due to their excellent features of high ion exchange capacity and absorbency. The aim of this study was to assess the effect of heavy metal uptake of one purified (PCT) and two non-purified clinoptilolite tuffs (NPCT1 and NPCT2) in aqueous solutions on monovalent ions Ni+, Cd+, Cs+, Ba+, Tl+, and Pb+. Experiments were furthermore carried out in artificial gastric and intestinal fluids to mimic human digestion and compare removal efficiencies of the adsorbent materials as well as release characteristics in synthetic gastric (SGF) and intestinal fluids (SIF). Batch experiments show low sorption capacities for Ni+ and Cd+ for all studied materials; highest affinities were found for Ba+ (99–100%), Pb+ (98–100%), Cs+ (97–98%), and Tl+ (96%), depending on the experimental setup for the PCT. For the adsorption experiments with SGF, highest adsorption was observed for the PCT for Pb+, with an uptake of 99% of the lead content. During artificial digestion, it was proven that the PCT did not release Ba+ cations into solution, whereas 13574 ng·g−1 and 4839 ng·g−1 of Ba+ were measured in the solutions with NPCT1 and NPCT2, respectively. It was demonstrated that the purified clinoptilolite tuff is most effective in remediating heavy-metal-polluted water, particularly during artificial digestion (99% of Pb+, 95% of Tl+, 93% of Ba+). In addition, it was shown that the released amount of bound heavy metal ions (e.g., barium) from the non-purified clinoptilolite tuffs into the intestinal fluids was significantly higher compared to the purified product.

Comparative transcriptome analysis revealed omnivorous adaptation of the small intestine of Melinae

As the main digestive organ, the small intestine plays a vital role in the digestion of animals. At present, most of the research on animal feeding habits focuses on carnivores and herbivores. However, the mechanism of feeding and digestion in omnivores remains unclear. This study aims to reveal the molecular basis of the omnivorous adaptive evolution of Melinae by comparing the transcriptome of the small intestines of Asian Badgers (Meles leucurus) and Northern Hog Badgers (Arctonyx albogularis). We obtained high-quality small intestinal transcriptome data from these two species. Key genes and signalling pathways were analysed through Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and other databases. Research has mainly found that orthologous genes related to six enzymes have undergone adaptive evolution. In addition, the study also found three digestion-related pathways (cGMP-PKG, cAMP, and Hippo). They are related to the digestion and absorption of nutrients, the secretion of intestinal fluids, and the transport of food through the small intestine, which may help omnivorous animals adapt to an omnivorous diet. Our study provides insight into the adaptation of Melinae to omnivores and affords a valuable transcriptome resource for future research.

Novel Yeasts Producing High Levels of Conjugated Linoleic Acid and Organic Acids in Fermented Doughs

Traditional fermented foods are obtained by a complex consortium of autochthonous microorganisms producing a wide variety of bioactive compounds, thus representing a reservoir of strains with new functional properties. Here, doughs obtained using five different wholegrain flours were singly fermented with selected yeast strains, which were evaluated for their functional traits. Lactate, volatile fatty acids and conjugated linoleic acid isomers produced by fermented doughs were detected by HPLC, while dough anti-inflammatory capacity was measured on human peripheral blood mononuclear cells by flow cytometry. Yeast potential probiotic activity was assessed by evaluating their resistance to simulated gastric and intestinal fluids. For the first time we report evidence of yeast strains producing high levels of the conjugated linoleic acid (CLA) isomer CLA 10-12tc and propionic acid, which are known for their specific health benefits. Moreover, such yeast strains showed an anti-inflammatory capacity, as revealed by a significantly decreased production of the strongly pro-inflammatory cytokine IL-1β. All our Saccharomyces cerevisiae strains were remarkably resistant to simulated gastric and intestinal fluids, as compared to the commercial probiotic strain. The two strains S. cerevisiae IMA D18Y and L10Y showed the best survival percentage. Our novel yeast strains may be exploited as valuable functional starters for the industrial production of cereal-based innovative and health-promoting fermented foods.

Bioaccessibility and Gut Metabolism of Free and Melanoidin-Bound Phenolic Compounds From Coffee and Bread

The aim of this study is to investigate the bioaccessibility and gut metabolism of free and melanoidin-bound phenolic compounds from coffee and bread. Phenolics from coffee were predominantly found in free forms (68%, mainly chlorogenic acids), whereas those from bread were mostly bound to melanoidins (61%, mainly ferulic acid). Bioacessibility of coffee total free phenolics slightly decreased during simulated digestion (87, 86, and 82% after the oral, gastric, and intestinal steps, respectively), with caffeoylquinic acids being isomerized and chlorogenic acids being partially hydrolyzed to the corresponding hydroxycinnamic acids. Bioacessibility of bread total free phenolics decreased during simulated digestion (91, 85, and 67% after the oral, gastric, and intestinal steps, respectively), probably related to complexation with the proteins in simulated gastric and intestinal fluids. Upon gut fermentation, the bioaccessibility of total free phenolics from both coffee and bread decreased, mainly after the first 4 h (56 and 50%, respectively). Caffeic and ferulic acids were the predominant metabolites found during coffee and bread gut fermentation, respectively. Melanoidin-bound phenolics from coffee and bread were progressively released after the gastric and intestinal steps, probably due to hydrolysis caused by the acidic conditions of the stomach and the action of pancreatin from the intestinal fluid. The bioaccessibilities of all phenolics from coffee and bread melanoidins after the gastric and intestinal steps were, on average, 11 and 26%, respectively. During gut fermentation, phenolics bound to both coffee and bread melanoidins were further released by the gut microbiota, whereas those from coffee were also metabolized. This difference could be related to the action of proteases on melanoproteins during gastrointestinal digestion, probably anticipating phenolics release. Nevertheless, bioaccessibilities of melanoidin-bound phenolics reached maximum values after gut fermentation for 24 h (50% for coffee and 51% for bread). In conclusion, the bioaccessibilities of coffee and bread free phenolics during simulated digestion and gut fermentation were remarkably similar, and so were the bioaccessibilities of coffee and bread melanoidin-bound phenolics.

Evaluation of cannabidiol’s inhibitory effect on alpha-glucosidase and its stability in simulated gastric and intestinal fluids

Objective Cannabidiol (CBD) has been reported to have anti-diabetic effects in pre-clinical and clinical studies but its inhibitory effects on α-glucosidase, a carbohydrate hydrolyzing enzyme, remain unknown. Herein, we evaluated CBD’s inhibitory effects on α-glucosidase using in vitro assays and computational studies. Methods CBD’s inhibitory effect on α-glucosidase activity was evaluated in a yeast enzymatic assay and by molecular docking. The stability of CBD in simulated gastric and intestinal fluids was evaluated by high-performance liquid chromatography analyses. Results CBD, at 10, 19, 38, 76, 152, 304, 608, and 1216 μM, inhibited α-glucosidase activity with inhibition of 17.1, 20.4, 48.1, 56.6, 59.1, 63.7, 74.1, and 95.4%, respectively. Acarbose, the positive control, showed a comparable inhibitory activity (with 85.1% inhibition at 608 μM). CBD’s inhibitory effect on α-glucosidase was supported by molecular docking showing binding energy (-6.39 kcal/mol) and interactions between CBD and the α-glucosidase protein. CBD was stable in simulated gastric and intestinal fluids for two hours (maintained ≥ 90.0%). Conclusions CBD showed moderate inhibitory effect against yeast α-glucosidase activity and was stable in gastric and intestinal fluids. However, further studies on CBD’s anti-α-glucosidase effects using cellular and in vivo models are warranted to support its potential application for the management of type II diabetes mellitus.

Determination of Honey Varietals’ Impact on Bifidobacterium animalis ssp lactis Survivability in Commercial Yogurt Through Simulated In Vitro Digestion

Objectives Consumption of yogurt containing the probiotic Bifidobacterium animalis lactis DN-173 010/CNCM I-2494 (B. animalis) improves digestive health and quality of life in adults. To optimize the benefits of this probiotic, we aimed to test our hypothesis that yogurt with honey would increase the survivability of B. animalis under simulated gastrointestinal tract digestion conditions. Methods Phase 1 tested four honey varietals (alfalfa, buckwheat, clover, and orange) at a final concentration of 20% w/w in yogurt containing B. animalis. Undiluted yogurt and yogurt with added sucrose or water (20% w/w) were included as control treatments. Phase 2 assessed clover honey at final concentrations of 20, 14, 10, 9, 8, 6, 4% w/w. Yogurt samples were subjected to in vitro simulated oral, gastric, and intestinal digestion using simulated salivary, gastric, and intestinal fluids, respectively. At four time points—pre-digestion, and after each phase of digestion (oral, gastric, intestinal)—probiotic cells were enumerated first by spread plating on MRS agar and incubated for 5 h at 37°C under anaerobic conditions. Then, plates were overlaid with MRS supplemented with lithium chloride and sodium propionate and incubated an additional 67 h prior to quantification of the probiotic colony forming units (CFU). Results Phase 1 demonstrated similar probiotic counts between honey varietals and controls after exposure to oral and gastric simulated fluids (< 1 Log CFU/g of probiotic reduction after gastric phase). There was comparable probiotic survival after the simulated intestinal phase for yogurt with the alfalfa, buckwheat, and orange honey varietals relative to control yogurt treatments. However, higher B. animalis survivability was observed in yogurt with clover honey after exposure to simulated intestinal fluids (∼3.5 Log CFU/g reduction) compared to all control treatments (∼5.5 Log CFU/g reduction, P ˂ 0.05). Phase 2 revealed that 20%, 14% and 10% w/w clover honey similarly supported B. animalis survivability after exposure to simulated intestinal fluids. Conclusions These results demonstrated that clover honey increased B. animalis survivability in yogurt during in vitro digestion when provided at doses equivalent to 1 to 2 tablespoons per serving (170g) of yogurt. Funding Sources The National Honey Board.