Cranberry Polyphenols and Prevention against Urinary Tract Infections: New Findings Related to the Integrity and Functionality of Intestinal and Urinary Barriers.

This work seeks to generate new knowledge about the mechanisms underlying the protective effects of cranberry against urinary tract infections (UTI). Using Caco-2 cells grown in Transwell inserts as an intestinal barrier model, we found that a cranberry-derived digestive fluid (containing 135 ± 5 mg of phenolic compounds/L) increased transepithelial electrical resistance with respect to control (ΔTEER = 54.5 Ω cm2) and decreased FITC-dextran paracellular transport by about 30%, which was related to the upregulation of the gene expression of tight junction (TJ) proteins (i.e., occludin, zonula occludens-1 [ZO-1], and claudin-2) (∼3-4-fold change with respect to control for claudin-2 and ∼2-3-fold for occludin and ZO-1). Similar protective effects, albeit to a lesser extent, were observed when Caco-2 cells were previously infected with uropathogenic Escherichia coli (UPEC). In a urinary barrier model comprising T24 cells grown in Transwell inserts and either noninfected or UPEC-infected, treatments with the cranberry-derived phenolic metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and phenylacetic acid (PAA) (250 µM) also promoted favorable changes in barrier integrity and permeability. In this line, incubation of noninfected T24 cells with these metabolites induced positive regulatory effects on claudin-2 and ZO-1 expression (∼3.5- and ∼2-fold change with respect to control for DOPAC and ∼1.5- and >2-fold change with respect to control for PAA, respectively). Overall, these results suggest that the protective action of cranberry polyphenols against UTI might involve molecular mechanisms related to the integrity and functionality of the urothelium and intestinal epithelium.

Simulated gastrointestinal digestion of beer using the simgi® model. Investigation of colonic phenolic metabolism and impact on human gut microbiota.

Beer is a source of bioactive compounds, mainly polyphenols, which can reach the large intestine and interact with colonic microbiota. However, the effects of beer consumption in the gastrointestinal function have scarcely been studied. This paper reports, for the first time, the in vitro digestion of beer and its impact on intestinal microbiota metabolism. Three commercial beers of different styles were subjected to gastrointestinal digestion using the simgi® model, and the digested fluids were further fermented in triplicate with faecal microbiota from a healthy volunteer. The effect of digested beer on human gut microbiota was evaluated in terms of microbial metabolism (short-chain fatty acids (SCFAs) and ammonium ion), microbial diversity and bacterial populations (plate counting and 16S rRNA gene sequencing). Monitoring beer polyphenols through the different digestion phases showed their extensive metabolism, mainly at the colonic stage. In addition, a higher abundance of taxa related to gut health, especially Bacteroides, Bifidobacterium, Mitsuokella and Succinilasticum at the genus level, and the Ruminococcaceae and Prevotellaceae families were found in the presence of beers. Regarding microbial metabolism, beer feeding significantly increased microbial SCFA production (mainly butyric acid) and decreased ammonium content. Overall, these results evidence the positive actions of moderate beer consumption on the metabolic activity of colonic microbiota, suggesting that the raw materials and brewing methods used may affect the beer gut effects.

Activity of Microbial-Derived Phenolic Acids and Their Conjugates against LPS-Induced Damage in Neuroblastoma Cells and Macrophages.

The aim of this study was to investigate whether microbial-derived phenolic acids, 3,4-dihydroxyphenylacetic (DHPA), protocatechuic acid (PCA), and dihydrocaffeic acid (DHCFA) and their conjugated forms (DHCFA 3-O-sulfate and DHCFA 3-O-ß-D-glucuronide), exhibit protective effects against neuroinflammation and oxidative stress. Experiments were performed on human neuronal SH-SY5Y cells stimulated with bacterial lipopolysaccharide (LPS) and tert-butyl hydroperoxide (tBHP). Anti-inflammatory activity in terms of pro-inflammatory cytokine production was also evaluated in LPS-stimulated RAW 264.7 macrophages as a reactive microglial model. Treatment of the SH-SY5Y cells with the free phenolic acids, as well as with the conjugated metabolites, at physiologically concentrations (1, 10 and 50 µM), resulted in increased cell viability of LPS- and tBHP-stimulated cells. Phenolic metabolites and, especially, the conjugated derivatives also protected neuronal cells through significant attenuation of inflammation by decreasing ROS levels. Furthermore, the conjugated and microbial-derived phenolic metabolites significantly inhibited the secretion of proinflammatory cytokines (TNF-α, IL-6, and IL-8) in LPS-stimulated macrophages. Among the phenolic metabolites tested, different efficacies were observed, with the glucuronide form standing out. Overall, these results suggest, for the first time, that conjugated derivatives of phenolic acids seem to be more effective at protecting neurons from inflammation damage and oxidative stress. Further in vivo studies are warranted.

Simulated gastrointestinal digestion of cranberry polyphenols under dynamic conditions. Impact on antiadhesive activity against uropathogenic bacteria.

This study is the first dynamic simulation of gastrointestinal digestion of cranberry polyphenols [1 g cranberry extract per day (206.2 mg polyphenols) for 18 days]. Samples from the simulated ascending, transverse, and descending colon of the dynamic gastrointestinal simulator simgi® were analyzed. Results showed that 67% of the total cranberry polyphenols were recovered after simulated gastrointestinal digestion. Specifically, benzoic acids, hydroxycinnamic acids, phenylpropionic acids, phenylacetic acids, and simple phenols were identified. Cranberry feeding modified colonic microbiota composition of Enterococcaceae population significantly. However, increments in microbial-derived short-chain fatty acids, particularly in butyric acid, were observed. Finally, the simgi® effluent during cranberry feeding showed significant antiadhesive activity against uropathogenic Escherichia coli (13.7 ± 1.59 % of inhibition). Understanding the role that gut microbiota plays in cranberry metabolism could help to elucidate its interaction with the human body and explain cranberry protective effects against urinary tract infections.

A Binary Logistic Regression Model as a Tool to Predict Craft Beer Susceptibility to Microbial Spoilage.

Beer spoilage caused by microorganisms, which is a major concern for brewers, produces undesirable aromas and flavors in the final product and substantial financial losses. To address this problem, brewers need easy-to-apply tools that inform them of beer susceptibility to the microbial spoilage. In this study, a growth/no growth (G/NG) binary logistic regression model to predict this susceptibility was developed. Values of beer physicochemical parameters such as pH, alcohol content (% ABV), bitterness units (IBU), and yeast-fermentable extract (% YFE) obtained from the analysis of twenty commercially available craft beers were used to prepare 22 adjusted beers at different levels of each parameter studied. These preparations were assigned as a first group of samples, while 17 commercially available beers samples as a second group. The results of G/NG from both groups, after artificially inoculating with one wild yeast and different lactic acid bacteria (LAB) previously adapted to grow in a beer-type beverage, were used to design the model. The developed G/NG model correctly classified 276 of 331 analyzed cases and its predictive ability was 100% in external validation. This G/NG model has good sensitivity and goodness of fit (87% and 83.4%, respectively) and provides the potential to predict craft beer susceptibility to microbial spoilage.

Gastrointestinal Digestion of a Grape Pomace Extract: Impact on Intestinal Barrier Permeability and Interaction with Gut Microbiome.

Grape pomace (GP) is a winemaking by-product rich in polyphenols and fibre. Supplementation with GP extracts has shown potential benefits against oxidative stress- and inflammation-related pathologies. As a new nutritional target, this paper explores the impact of the ingestion of a grape pomace extract on intestinal barrier functionality. A GP extract was sequentially subjected to gastrointestinal and colonic digestion using the dynamic gastrointestinal simulator (simgi®). This generated two simulated fluids: intestinal-digested extract (IDE) and colonic-digested extract (CDE). The effects of these two fluids on paracellular permeability and the expression of tight junction (TJ) proteins (i.e., zonula occludens-1 (ZO-1) and occludin) were assessed in Caco-2-cell monolayers grown in Transwell® inserts. The IDE fluid significantly (p < 0.001) reduced the paracellular transport of FITC-dextran with respect to the control, whereas no significant differences (p > 0.05) were found for CDE, which could be due, at least partially, to the pro-leaky effect of the colonic digestion medium. Accordant slight increases in the mRNA levels of both ZO-1 and occludin were observed for IDE, but without statistical significance. Additionally, the colonic fermentation of the GP extract promoted the production of short-chain fatty acids (SCFA) and phenolic metabolites and led to changes in the relative abundance of some bacteria that might affect paracellular permeability. Overall, this paper reports first trends about the effects of grape pomace extracts on intestinal permeability that would require further confirmation in future experiments.

Pectinatus spp. - Unpleasant and recurrent brewing spoilage bacteria.

Traditionally, beer has been recognised as a beverage with high microbiological stability because of the hostile growth environment posed by beer and increasing attention being paid to brewery hygiene. However, the microbiological risk has increased in recent years because of technological advances toward reducing oxygen in beers, besides the increase in novel beer styles production, such as non-pasteurised, flash pasteurised, cold sterilised, mid-strength, and alcoholic-free beer, that are more prone to spoilage bacteria. Moreover, using innovative beer ingredients like fruits and vegetables is an added cause of microbial spoilage. To maintain quality and good brand image, beer spoilage microorganisms are a critical concern for breweries worldwide. Pectinatus and Megasphaera are Gram-negative bacteria mostly found in improper brewing environments, leading to consumer complaints and financial losses. Because of the lack of compiled scientific knowledge on Pectinatus spoilage ability, this review provides a comprehensive overview of the occurrence, survival mechanisms, and the factors affecting beer spoilage Pectinatus species in the brewing process.

Beer spoilage lactic acid bacteria from craft brewery microbiota: Microbiological quality and food safety.

Craft beer is more susceptible to microbial spoilage because it does not have a pasteurization or filtration process, with lactic acid bacteria (LAB) being the most common beer spoilage microorganism. The aim of this study was to isolate LAB in a craft brewery and their characterization from a food safety and microbiological quality perspective, with a special focus on their abilities to produce biogenic amines (BA) and spoil the beer. The results of 60 monitored points inside the craft brewery showed that LAB associated with the craft brewing processes belonged to Lactobacillus, Pediococcus, and Leuconostoc genera, and most of them were detected in the filling area, which can lead to secondary contamination. Two isolates of L. brevis showed the most significant beer spoilage ability because they could grow in more acidic conditions, at a higher hop and alcohol content, and they displayed horA, horC, and hitA genes, which spoiled the vast majority of the tested beers. In addition, the aforementioned L. brevis isolates showed the highest BA production.

Glutathione-Stabilized Silver Nanoparticles: Antibacterial Activity against Periodontal Bacteria, and Cytotoxicity and Inflammatory Response in Oral Cells.

Silver nanoparticles (AgNPs) have been proposed as new alternatives to limit bacterial dental plaque because of their antimicrobial activity. Novel glutathione-stabilized silver nanoparticles (GSH-AgNPs) have proven powerful antibacterial properties in food manufacturing processes. Therefore, this study aimed to evaluate the potentiality of GSH-AgNPs for the prevention/treatment of oral infectious diseases. First, the antimicrobial activity of GSH-AgNPs against three oral pathogens (Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans) was evaluated. Results demonstrated the efficiency of GSH-AgNPs in inhibiting the growth of all bacteria, especially S. mutans (IC50 = 23.64 µg/mL, Ag concentration). Second, GSH-AgNPs were assayed for their cytotoxicity (i.e., cell viability) toward a human gingival fibroblast cell line (HGF-1), as an oral epithelial model. Results indicated no toxic effects of GSH-AgNPs at low concentrations (≤6.16 µg/mL, Ag concentration). Higher concentrations resulted in losing cell viability, which followed the Ag accumulation in cells. Finally, the inflammatory response in the HGF-1 cells after their exposure to GSH-AgNPs was measured as the production of immune markers (interleukins 6 and 8 (IL-6 and IL-8) and tumor necrosis factor-alpha (TNF-α)). GSH-AgNPs activates the inflammatory response in human gingival fibroblasts, increasing the production of cytokines. These findings provide new insights for the use of GSH-AgNPs in dental care and encourage further studies for their application.

Moderate Wine Consumption Reduces Faecal Water Cytotoxicity in Healthy Volunteers.

There are some studies that suggest that moderate consumption of wine, as part of a healthy and balanced diet, has a favourable effect on intestinal health. This study evaluates the effect of moderate wine consumption on faecal water (FW) cytotoxicity as a parameter of gut health. To that end, faecal samples before and after a red wine intervention study (250 mL of wine/day, 4 weeks) in healthy volunteers (n = 8) and in a parallel control group (n = 3) were collected and assayed for in vitro FW cytotoxicity. Two reference compounds, phenol and p-cresol, were used for assessing the cytotoxicity assays using two colon epithelial cell lines (HT-29 and HCT 116) and different assay conditions (FW dilution and incubation time). For the two cell lines and all assay conditions, the means of percentage cell viability were higher (lower cytotoxicity) for samples collected after the red wine intervention than for those collected before, although significant (p < 0.05) differences were only found in certain assay conditions for both cell lines. Significant positive correlations between the percentage cell viability and the contents of some faecal metabolites (short-chain fatty acids (SCFA) and phenolic acids (PA)) were found for the more resistant cell line (HCT 116), suggesting that the reduction in FW cytotoxicity observed after moderate red wine consumption was related to the production of microbial-derived metabolites such as SCFA and PA, whose faecal contents have been shown to increase after wine consumption. FW cytotoxicity can be deemed as a holistic biomarker that involves diet, gut microbiota and host.

Cranberry Polyphenols and Prevention against Urinary Tract Infections: Relevant Considerations.

Cranberry (Vaccinium macrocarpon) is a distinctive source of polyphenols as flavonoids and phenolic acids that has been described to display beneficial effects against urinary tract infections (UTIs), the second most common type of infections worldwide. UTIs can lead to significant morbidity, especially in healthy females due to high rates of recurrence and antibiotic resistance. Strategies and therapeutic alternatives to antibiotics for prophylaxis and treatment against UTIs are continuously being sought after. Different to cranberry, which have been widely recommended in traditional medicine for UTIs prophylaxis, probiotics have emerged as a new alternative to the use of antibiotics against these infections and are the subject of new research in this area. Besides uropathogenic Escherichia coli (UPEC), the most common bacteria causing uncomplicated UTIs, other etiological agents, such as Klebsiellapneumoniae or Gram-positive bacteria of Enterococcus and Staphylococcus genera, seem to be more widespread than previously appreciated. Considerable current effort is also devoted to the still-unraveled mechanisms that are behind the UTI-protective effects of cranberry, probiotics and their new combined formulations. All these current topics in the understanding of the protective effects of cranberry against UTIs are reviewed in this paper. Further progresses expected in the coming years in these fields are also discussed.

Some New Findings Regarding the Antiadhesive Activity of Cranberry Phenolic Compounds and Their Microbial-Derived Metabolites against Uropathogenic Bacteria.

Findings concerning the antiadhesive activity of cranberry phenolic compounds and their microbial-derived metabolites against Gram-negative ( Escherichia coli ATCC 53503 and DSM 10791) and Gram-positive ( Enterococcus faecalis 04-1) bacteria in T24 cells are reported. A-Type procyanidins (A2 and cinnamtannin B-1) exhibited antiadhesive activity (at concentrations ≥250 µM), a feature that was not observed for B-type procyanidins (B2). The metabolites hippuric acid and α-hydroxyhippuric acid also showed effective results at concentrations ≥250 µM. With regard to conjugated metabolites, sulfation seemed to increase the antiadhesive activity of cranberry-derived metabolites as 3-(3,4-dihydroxyphenyl)propionic acid 3- O-sulfate presented active results, unlike its corresponding nonsulfated form. In contrast, methylation decreased antiadhesive activity as 3,4-dihydroxyphenylacetic acid was found to be active but not its corresponding methylated form (4-hydroxy-3-methoxyphenylacetic acid). As a whole, this work sustains the antiadhesive activity of cranberry-derived metabolites as one of the mechanisms involved in the beneficial effects of cranberries against urinary tract infections.

Antibacterial Activity of Glutathione-Stabilized Silver Nanoparticles Against Campylobacter Multidrug-Resistant Strains.

Campylobacter is the leading cause of bacterial diarrheal disease worldwide. Although most episodes of campylobacteriosis are self-limiting, antibiotic treatment is usually needed in patients with serious enteritis, and especially in childrens or the elderly. In the last years, antibiotic resistance in Campylobacter has become a major public health concern and a great interest exists in developing new antimicrobial strategies for reducing the impact of this food-borne pathogen on human health. Among them, the use of silver nanoparticles as antibacterial agents has taken on increased importance in the field of medicine. The aim of the present study was to evaluate the antimicrobial effectiveness of glutathione-stabilized silver nanoparticles (GSH-Ag NPs) against multidrug resistant (MDR) Campylobacter strains isolated from the chicken food chain (FC) and clinical patients (C). The results obtained showed that GSH-Ag NPs were highly effective against all MDR Campylobacter strains tested. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were in a range from 4.92 to 39.4 µg/mL and 9.85 to 39.4 µg/mL, respectively. Cytotoxicity assays were also assessed using human intestinal HT-29, Caco-2, and CCD-18 epithelial cells. Exposure of GSH-Ag NPs to intestinal cells showed a dose-dependent cytotoxic effect in all cell lines between 9.85 and 39.4 µg/mL. More than 60% of the tested Campylobacter strains were susceptible to GSH-Ag NPs concentrations ≤ 9.85 µg/mL, suggesting that practical inhibitory levels could be reached at low GSH-Ag NPs concentrations. Further works are needed with the purpose to evaluate the practical implications of the toxicity studies and to know more about other attributes linked to the biological compatibility. This behavior makes GSH-Ag NPs as a promising tool for the design of novel antibacterial agents for controlling Campylobacter.

Inhibition of Oral Pathogens Adhesion to Human Gingival Fibroblasts by Wine Polyphenols Alone and in Combination with an Oral Probiotic.

Several benefits have been described for red wine polyphenols and probiotic strains in the promotion of colonic metabolism and health. On the contrary, knowledge about their role in the management of oral health is still scarce. In this work, the antiadhesive capacity of selected red wine polyphenols and oenological extracts against the oral pathogens Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans in an in vitro model of human gingival fibroblasts has been explored as well as their complementary action with the candidate oral probiotic Streptococcus dentisani. Results highlighted the antiadhesive capacity of caffeic and p-coumaric acids as well as grape seed and red wine oenological extracts. Both, caffeic and p-coumaric acids increased their inhibition potential against S. mutans adhesion when combined with S. dentisani. Additionally, UHPLC-MS/MS analysis demonstrated the oral metabolism of wine phenolics due to both, cellular and bacterial activity.

Assessment of probiotic properties in lactic acid bacteria isolated from wine.

Probiotic properties are highly strain-dependent but rarely studied in enological lactic acid bacteria (LAB). In this study, the probiotic features of 11 strains of Lactobacillus spp., Pediococcus spp., and Oenococcus oeni, including saliva and acid resistance, bile tolerance and exopolysaccharides' production, were investigated. The assays included two probiotic reference strains (Lactobacillus plantarum CLC 17 and Lactobacillus fermentum CECT5716). The Lactobacillus and Pediococcus strains showed high resistance to lysozyme (>80% resistance to 100 mg/L of lysozyme under conditions simulating the in vivo dilution by saliva) and were capable of surviving at low pH values (pH 1.8) and bile salts, suggesting good adaptation of the wine strains to gastrointestinal conditions. The ability of the strains to adhere to the intestinal mucosa and the inhibition of the adhesion of Escherichia coli to human intestinal cells were also evaluated. Adhesion levels of enological LAB to Caco-2 cells varied from 0.37% to 12.2%, depending on the strain. In particular, Pediococcus pentosaceus CIAL-86 showed a high percentage of adhesion to intestinal cells (>12%), even higher than that shown by the probiotic reference strains, and a high anti-adhesion activity against E. coli CIAL-153 (>30%), all of which support this wine LAB strain as a potential probiotic.