Effect of acute meal and long-term intake of a Mediterranean Diet providing different amounts of carbohydrates on physical performance and biomarkers in non-professional strength athletes.

The study aims to evaluate the effect of an acute meal and long-term intake of Mediterranean Diet (MD) on different parameters such as strength, physical performance, body composition and blood markers in a group of non-professional athletes who practice a strength activity. Thirteen volunteers completed two 8-week dietary interventions in a randomised, cross-over design. Also an acute study was performed. Subjects received a MD High in carbohydrates, characterised by at least five portions of pasta/week and an average 55-60% of daily energy derived from carbohydrates, versus an MD reduced in carbohydrates, with less than two portions of pasta/week and an average of 40-45% of daily energy provided by carbohydrates. Mainly, data did not show significant differences for the parameters analysed, except for Elbow Flexor maximum voluntary contraction (p = .039). Results enlighten that increasing total carbohydrates intake, as typically in the MD, does not negatively affect physical performance, body composition and strength.

Screening of Mpro Protease (SARS-CoV-2) Covalent Inhibitors from an Anthocyanin-Rich Blueberry Extract Using an HRMS-Based Analytical Platform.

BACKGROUND: The viral main protease (Mpro) of SARS-CoV-2 has been recently proposed as a key target to inhibit virus replication in the host. Therefore, molecules that can bind the catalytic site of Mpro could be considered as potential drug candidates in the treatment of SARS-CoV-2 infections. Here we proposed the application of a state-of-the-art analytical platform which combines metabolomics and protein structure analysis to fish-out potential active compounds deriving from a natural matrix, i.e., a blueberry extract. METHODS: The experiments focus on finding MS covalent inhibitors of Mpro that contain in their structure a catechol/pyrogallol moiety capable of binding to the nucleophilic amino acids of the enzyme's catalytic site. RESULTS: Among the potential candidates identified, the delphinidin-3-glucoside showed the most promising results. Its antiviral activity has been confirmed in vitro on Vero E6 cells infected with SARS-CoV-2, showing a dose-dependent inhibitory effect almost comparable to the known Mpro inhibitor baicalin. The interaction of delphinidin-3-glucoside with the Mpro pocket observed was also evaluated by computational studies. CONCLUSIONS: The HRMS analytical platform described proved to be effective in identifying compounds that covalently bind Mpro and are active in the inhibition of SARS-CoV-2 replication, such as delphinidin-3-glucoside.

Red raspberry (Rubus idaeus) preserves intestinal barrier integrity and reduces oxidative stress in Caco-2 cells exposed to a proinflammatory stimulus.

Growing evidence showed the capacity of (poly)phenols to exert a protective role on intestinal health. Nevertheless, the existing findings are still heterogeneous and the underlying mechanisms remain unclear. This study investigated the potential benefits of a red raspberry (Rubus idaeus) powder on the integrity of the intestinal barrier, focusing on its ability to mitigate the effects of tumor necrosis factor-α (TNF-α)-induced intestinal permeability. Human colorectal adenocarcinoma cells (i.e., Caco-2 cells) were used as a model to assess the impact of red raspberry on intestinal permeability, tight junction expression, and oxidative stress. The Caco-2 cells were differentiated into polarized monolayers and treated with interferon-γ (IFN-γ) (10 ng mL-1) for 24 hours, followed by exposure to TNF-α (10 ng mL-1) in the presence or absence of red raspberry extract (1-5 mg mL-1). The integrity of the intestinal monolayer was evaluated using transepithelial electrical resistance (TEER) and fluorescein isothiocyanate-dextran (FITC-D) efflux assay. Markers of intestinal permeability (claudin-1, occludin, and zonula occludens-1 (ZO-1)) and oxidative stress (8-hydroxy-2-deoxyguanosine (8-OHdG) and protein carbonyl) were assessed using ELISA kits. Treatment with red raspberry resulted in a significant counteraction of TEER value loss (41%; p < 0.01) and a notable reduction in the efflux of FITC-D (-2.5 times; p < 0.01). Additionally, red raspberry attenuated the levels of 8-OHdG (-48.8%; p < 0.01), mitigating the detrimental effects induced by TNF-α. Moreover, red raspberry positively influenced the expression of the integral membrane protein claudin-1 (+18%; p < 0.01), an essential component of tight junctions. These findings contribute to the growing understanding of the beneficial effects of red raspberry in the context of the intestinal barrier. The effect of red raspberry against TNF-α-induced intestinal permeability observed in our in vitro model suggests, for the first time, its potential as a dietary strategy to promote gastrointestinal health.

The MED_EAT-IT approach: A modelling study to develop feasible, sustainable and nutritionally targeted dietary patterns based on the Planetary health diet.

In 2019, the EAT-Lancet Commission introduced the Planetary Health Diet (PHD), a guide for creating 2500 kcal/day country-specific sustainable diets that promote health while reducing the environmental impact associated with food systems. The PHD was previously adapted to the Italian food context, resulting in the EAT-IT dietary pattern. However, this adaptation revealed several challenges in terms of nutritional adequacy, feasibility, and environmental impact. This study reports on strategies to improve the previous pattern and align it more closely with the Mediterranean Diet, resulting in the MED_EAT-IT pattern. The study also explores feasible strategies for adapting this pattern to different energy targets, enhancing its scalability and promoting personalized approaches. For the optimization of this pattern, a specific calculation tool was developed to introduce variation to the pattern, considering realistic and feasible serving sizes and frequency of consumption. This tool integrates a defined food ontology, food composition data, and two environmental impact metrics (Carbon and Water Footprint). To optimize nutritional adequacy, several adaptations of the amount within the different food groups were made, for instance by increasing cereals and animal source by 25.5% kcal/day and 36.2% kcal/day respectively compared to EAT-IT. The resulting 2500 kcal/die pattern meets all nutritional requirements except for vitamin D and does not hamper the possibility to limit environmental impact (Carbon Footprint increased only by 12.2% but Water Footprint decreased by 6.3%). Lower energy targets were achieved by modulating amounts of the different food groups to ensure nutritional adequacy. The strategies and tools proposed here could aid in optimizing dietary plans, evaluating their potential for environmental impact reduction, and identifying issues that could hinder their adoption. Furthermore, the analyses carried out pave the way for the potential future development of new or improved foods that may contribute to the optimization of nutritional and environmental impact of diets.

Nutritional and Phytochemical Characterization of Freeze-Dried Raspberry (Rubus idaeus): A Comprehensive Analysis.

Several studies have highlighted the beneficial effects of consuming red raspberries on human health thanks to their high content of phytochemicals. However, the products used in these studies, both in the raw or freeze-dried form, were not fully characterized for nutrient and phytochemical composition. In this study, we aimed to determine the nutrient and non-nutrient compounds present in a freeze-dried red raspberry powder widely used by the food industry and consumers. The main sugars identified were fructose (12%), glucose (11%), and sucrose (11%). Twelve fatty acids were detected, with linoleic acid (46%), α-linolenic acid (20%), and oleic acid (15%) being the most abundant. Regarding micronutrients, vitamin C was the main hydro-soluble vitamin, while minerals, potassium, phosphorous, copper and magnesium were the most abundant, with concentrations ranging from 9 up to 96 mg/100 g, followed by manganese, iron and zinc, detected in the range 0.1-0.9 mg/100 g. Phytochemical analysis using UHPLC-DAD-HR-MS detection revealed the presence of Sanguiin H6 (0.4%), Lambertianin C (0.05%), and Sanguiin H-10 isomers (0.9%) as the main compounds. Among anthocyanins, the most representative compounds were cyanidin-3-sophoroside, cyanidin-3-glucoside and cyanidin-3-sambubioside. Our findings can serve as a reliable resource for the food industry, nutraceutical applications and for future investigations in the context of human health.

Effect of caper fruit (Capparis spinosa L.) consumption on liver enzymes, lipid profile, fasting plasma glucose, and weight loss. A systematic review and a preliminary meta-analysis of randomized controlled trials.

This systematic review and meta-analysis aimed to evaluate the overall effect of caper fruit on the modulation of glycemic, lipid profile, liver enzymes, and body mass. Google Scholar, PubMed, and Scopus were explored to collect relevant studies in the last 10 years. RCTs with caper fruit supplementation or consumption in different cohorts of subjects with non-alcoholic fatty liver disease (NAFLD), Type-2-Diabetes (T2D), metabolic syndrome, and hyperlipidemia were included in this systematic review with a mean intervention duration from 2 to 12 weeks. The outcomes measured in this meta-analysis were liver enzymes such as aspartate aminotransferase (AST) and alanine aminotransferase (ALT), the lipid profile represented by triglycerides, total cholesterol (TC) with LDL and HDL and also, weight, and fasting blood glucose. Five randomized controlled trials, which involved a total of 178 adults, were included. According to the results, caper fruit seems to decrease liver enzymes ALT -12.29 U/L [-24.47, -0.11], AST -2.20 U/L [-4.70, 0.31]. Furthermore, the lipid profile seems to improve with a decrease in triglycerides. -11.89 mg/dL [-33.73, 9.95], LDL -4.80 mg/dL [-16.34, 6.74], HDL 0.72 mg/dL [0.10, 1.34], total cholesterol -7.83 mg/dL [-20.04, 4.38], FPG -17.93 [-42.66, 6.79], weight -1.00 kg [-1.44, -0.56]. Significant modulations were found only for ALT, HDL, and weight. In conclusion, this systematic review and meta-analysis showed the paucity of data available on the topic while showing the potential role of caper fruit as a promising food for improving the liver-lipid profile axis in patients with metabolic syndrome and diabetes. Further studies are required to confirm these results.

Wild blueberry (V. angustifolium) improves TNFα-induced cell barrier permeability through claudin-1 and oxidative stress modulation in Caco-2 cells.

Increasing evidence links the impairment of intestinal permeability (IP), a feature of the intestinal barrier, to numerous dysmetabolic and dysfunctional conditions. Several host and environmental factors, including dietary factors, can negatively and/or positively affect IP. In this regard, polyphenol-rich foods including berries have been proposed as potential IP modulators. However, the exact mechanisms involved are not yet fully elucidated. The aim of the present study was to evaluate the effect of a wild blueberry (WB; V. angustifolium) powder, naturally rich in polyphenols, to affect Caco-2 cell monolayer permeability and to identify the potential mechanisms in modulating the IP process. Caco-2 cells were incubated with TNF-α (10 ng mL-1), as a pro-inflammatory stimulus, and supplemented for 24 hours with different concentrations (1 and 5 mg mL-1) of WB powder. The integrity of the intestinal cell monolayer was evaluated by measuring the transepithelial electrical resistance (TEER) and the paracellular transport of FITC-dextran. In addition, the production of the tight junction proteins, such as claudin-1 and occludin, as well as protein carbonyl and 8-hydroxy 2 deoxyguanosine, as oxidative stress markers, were quantified in the supernatant by ELISA kits. Overall, the treatment with WB powder (5 mg mL-1) mitigated the loss of Caco-2 cell barrier integrity, as documented by an increase in TEER and a reduction in FITC values. This modulation was accompanied by an upregulation of claudin-1 and a reduction of 8-OHdG. Conversely, no effect was documented for the lower concentration (1 mg mL-1) and the other IP markers, as well as oxidative stress markers analysed. In conclusion, our findings suggest a potential role of WB in the modulation of cell barrier integrity. This modulation process could be attributed to an increase in claudin-1 expression and a reduction in 8-OHdG. Further studies should be performed to corroborate the results obtained. In addition, since the effects were observed at doses of WB achievable with the diet, these findings should be substantiated also through in vivo approaches.

Healthy Eating for Elective Major Orthopedic Surgery: Quality, Quantity, and Timing.

Improvements to enhanced recovery pathways in orthopedic surgery are reducing the time that patients spend in the hospital, giving an increasingly vital role to prehabilitation and/or rehabilitation after surgery. Nutritional support is an important tenant of perioperative medicine, with the aim to integrate the patient's diet with food components that are needed in greater amounts to support surgical fitness. Regardless of the time available between the time of contemplation of surgery and the day of admission, a patient who eats healthy is reasonably more suitable for surgery than a patient who does not meet the daily requirements for energy and nutrients. Moreover, a successful education for healthy food choices is one possible way to sustain the exercise therapy, improve recovery, and thus contribute to the patient's long-term health. The expected benefits presuppose that the patient follows a healthy diet, but it is unclear which advice is needed to improve dietary choices. We present the principles of healthy eating for patients undergoing major orthopedic surgery to lay the foundations of rational and valuable perioperative nutritional support programs. We discuss the concepts of nutritional use of food, requirements, portion size, dietary target, food variety, time variables of feeding, and the practical indications on what the last meal to be consumed six hours before the induction of anesthesia may be together with what is meant by clear fluids to be consumed until two hours before. Surgery may act as a vital "touch point" for some patients with the health service and is therefore a valuable opportunity for members of the perioperative team to promote optimal lifestyle choices, such as the notion and importance of healthy eating not just for surgery but also for long-term health benefit.

DNA strand break levels in cryopreserved mononuclear blood cell lines measured by the alkaline comet assay: results from the hCOMET ring trial.

The comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, 10 laboratories used standardized cryopreservation and thawing procedures of monocytic (THP-1) or lymphocytic (TK6) cells. Samples were cryopreserved in small aliquots in 50% foetal bovine serum, 40% cell culture medium, and 10% dimethyl sulphoxide. Subsequently, cryopreserved samples were analysed by the standard comet assay on three occasions over a 3-year period. Levels of DNA strand breaks in THP-1 cells were increased (four laboratories), unaltered (four laboratories), or decreased (two laboratories) by long-term storage. Pooled analysis indicates only a modest positive association between storage time and levels of DNA strand breaks in THP-1 cells (0.37% Tail DNA per year, 95% confidence interval: -0.05, 0.78). In contrast, DNA strand break levels were not increased by cryopreservation in TK6 cells. There was inter-laboratory variation in levels of DNA strand breaks in THP-1 cells (SD = 3.7% Tail DNA) and TK6 reference sample cells (SD = 9.4% Tail DNA), whereas the intra-laboratory residual variation was substantially smaller (i.e. SD = 0.4%-2.2% Tail DNA in laboratories with the smallest and largest variation). In conclusion, the study shows that accumulation of DNA strand breaks in cryopreserved mononuclear blood cell lines is not a matter of concern.

Long-term cryopreservation of potassium bromate positive assay controls for measurement of oxidatively damaged DNA by the Fpg-modified comet assay: results from the hCOMET ring trial.

The formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with a little concurrent generation of DNA strand breaks. Eight laboratories used the same procedure for the treatment of monocytic THP-1 cells with potassium bromate (0, 0.5, 1.5, and 4.5 mM) and subsequent cryopreservation in a freezing medium consisting of 50% foetal bovine serum, 40% RPMI-1640 medium, and 10% dimethyl sulphoxide. The samples were analysed by the Fpg-modified comet assay three times over a 3-year period. All laboratories obtained a positive concentration-response relationship in cryopreserved samples (linear regression coefficients ranging from 0.79 to 0.99). However, there was a wide difference in the levels of Fpg-sensitive sites between the laboratory with the lowest (4.2% Tail DNA) and highest (74% Tail DNA) values in THP-1 cells after exposure to 4.5 mM KBrO3. In an attempt to assess sources of inter-laboratory variation in Fpg-sensitive sites, comet images from one experiment in each laboratory were forwarded to a central laboratory for visual scoring. There was high consistency between measurements of %Tail DNA values in each laboratory and the visual score of the same comets done in the central laboratory (r = 0.98, P < 0.001, linear regression). In conclusion, the results show that potassium bromate is a suitable positive comet assay control.

Can the Substitution of Milk with Plant-Based Drinks Affect Health-Related Markers? A Systematic Review of Human Intervention Studies in Adults.

The consumption of plant-based drinks (PBDs) in substitution for cow's milk (CM) is increasing due to concerns for human and planet health and animal welfare. The present review aims to analyze the main findings from intervention trials investigating the effect of PBDs in comparison with CM on markers of human health. Suitable articles published up to July 2022 were sourced from PubMed and Scopus databases. A total of 29 papers were collected, with 27 focusing on soy drinks (1 of which also evaluated the effects of an almond drink), while only 2 focused on rice drinks. Among studies focused on soy drinks, the most investigated factors were anthropometric parameters (n = 13), the lipid profile (n = 8), markers of inflammation and/or oxidative stress (n = 7), glucose and insulin responses (n = 6) and blood pressure (n = 4). Despite some evidence of a beneficial effect of PBDs, especially for the lipid profile, it was not possible to draw any overall conclusions due to some conflicting results. As well as the low number of studies, a wide heterogeneity was found in terms of the characteristics of subjects, duration and markers, which reduces the strength of the available results. In conclusion, further studies are needed to better elucidate the effects of substituting CM with PBDs, especially in the long term.

Visual comet scoring revisited: a guide to scoring comet assay slides and obtaining reliable results.

Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20%-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 different laboratories scored the comet images using a five-class scoring system. There is inter-investigator variation in the three training sets of comets (i.e. coefficient of variation (CV) = 9.7%, 19.8%, and 15.2% in training sets I-III, respectively). However, there is also a positive correlation of inter-investigator scoring in the three training sets (r = 0.60). Overall, 36% of the variation is attributed to inter-investigator variation and 64% stems from intra-investigator variation in scoring between comets (i.e. the comets in training sets I-III look slightly different and this gives rise to heterogeneity in scoring). Intra-investigator variation in scoring was also assessed by repeated analysis of the training sets by the same investigator. There was larger variation when the training sets were scored over a period of six months (CV = 5.9%-9.6%) as compared to 1 week (CV = 1.3%-6.1%). A subsequent study revealed a high inter-investigator variation when premade slides, prepared in a central laboratory, were stained and scored by investigators in different laboratories (CV = 105% and 18%-20% in premade slides with comets from unexposed and hydrogen peroxide-exposed cells, respectively). The results indicate that further standardization of visual scoring is desirable. Nevertheless, the analysis demonstrates that visual scoring is a reliable way of analysing DNA migration in comets.

Inter-laboratory variation in measurement of DNA damage by the alkaline comet assay in the hCOMET ring trial.

The comet assay is a simple and versatile method for measurement of DNA damage in eukaryotic cells. More specifically, the assay detects DNA migration from agarose gel-embedded nucleoids, which depends on assay conditions and the level of DNA damage. Certain steps in the comet assay procedure have substantial impact on the magnitude of DNA migration (e.g. electric potential and time of electrophoresis). Inter-laboratory variation in DNA migration levels occurs because there is no agreement on optimal assay conditions or suitable assay controls. The purpose of the hCOMET ring trial was to test potassium bromate (KBrO3) as a positive control for the formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay. To this end, participating laboratories used semi-standardized protocols for cell culture (i.e. cell culture, KBrO3 exposure, and cryopreservation of cells) and comet assay procedures, whereas the data acquisition was not standardized (i.e. staining of comets and image analysis). Segregation of the total variation into partial standard deviation (SD) in % Tail DNA units indicates the importance of cell culture procedures (SD = 10.9), comet assay procedures (SD = 12.3), staining (SD = 7.9) and image analysis (SD = 0.5) on the overall inter-laboratory variation of DNA migration (SD = 18.2). Future studies should assess sources of variation in each of these steps. On the positive side, the hCOMET ring trial demonstrates that KBrO3 is a robust positive control for the Fpg-modified comet assay. In conclusion, the hCOMET ring trial has demonstrated a high reproducibility of detecting genotoxic effects by the comet assay, but inter-laboratory variation of DNA migration levels is a concern.

Does the Mediterranean Diet Have Any Effect on Lipid Profile, Central Obesity and Liver Enzymes in Non-Alcoholic Fatty Liver Disease (NAFLD) Subjects? A Systematic Review and Meta-Analysis of Randomized Control Trials.

The effectiveness of the Mediterranean diet (MD) in non-alcoholic fatty liver disease (NAFLD) subjects has been evaluated in several randomized controlled trials (RCTs). This systematic review and meta-analysis aimed to evaluate the overall effects of MD intervention in a cohort of NAFLD patients targeting specific markers such as central obesity, lipid profile, liver enzymes and fibrosis, and intrahepatic fat (IHF). Google Scholar, PubMed, and Scopus were explored to collect relevant studies from the last 10 years. RCTs with NAFLD subjects were included in this systematic review with a mean intervention duration from 6 weeks to 1 year, and different intervention strategies, mainly including energy restriction MD (normal or low glycaemic index), low-fat MD with increased monounsaturated and polyunsaturated fatty acids, and increased exercise expenditure. The outcomes measured in this meta-analysis were gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), total cholesterol (TC), waist circumference (WC), and liver fibrosis. Ten randomized controlled trials, which involved a total of 737 adults with NAFLD, were included. According to the results, the MD seems to decrease the liver stiffness (kPa) by -0.42 (CI95% -0.92, 0.09) (p = 0.10) and significantly reduce the TC by -0.46 mg/dl (CI95% -0.55, -0.38) (p = 0.001), while no significant findings were documented for liver enzymes and WC among patients with NAFLD. In conclusion, the MD might reduce indirect and direct outcomes linked with NAFLD severity, such as TC, liver fibrosis, and WC, although it is important to consider the variations across trials. Further RCTs are necessary to corroborate the findings obtained and provide further evidence on the role of the MD in the modulation of other disorders related to NAFLD.

Garlic consumption in relation to colorectal cancer risk and to alterations of blood bacterial DNA.

PURPOSE: Garlic consumption has been inversely associated to intestinal adenoma (IA) and colorectal cancer (CRC) risk, although evidence is not consistent. Gut microbiota has been implied in CRC pathogenesis and is also influenced by garlic consumption. We analyzed whether dietary garlic influence CRC risk and bacterial DNA in blood. METHODS: We conducted a case-control study in Italy involving 100 incident CRC cases, 100 IA and 100 healthy controls matched by center, sex and age. We used a validated food frequency questionnaire to assess dietary habits and garlic consumption. Blood bacterial DNA profile was estimated using qPCR and16S rRNA gene profiling. We derived odds ratios (ORs) and the corresponding 95% confidence intervals (CIs) of IA and CRC according to garlic consumption from multiple conditional logistic regression. We used Mann-Whitney and chi-square tests to evaluate taxa differences in abundance and prevalence. RESULTS: The OR of CRC for medium/high versus low/null garlic consumption was 0.27 (95% CI = 0.11-0.66). Differences in garlic consumption were found for selected blood bacterial taxa. Medium/high garlic consumption was associated to an increase of Corynebacteriales order, Nocardiaceae family and Rhodococcus genus, and to a decrease of Family XI and Finegoldia genus. CONCLUSIONS: The study adds data on the protective effect of dietary garlic on CRC risk. Moreover, it supports evidence of a translocation of bacterial material to bloodstream and corroborates the hypothesis of a diet-microbiota axis as a mechanism behind the role of garlic in CRC prevention.

Berry Dietary Interventions in Metabolic Syndrome: New Insights.

Metabolic Syndrome (MetS) is characterized by a group of dysmetabolic conditions, including abdominal obesity, dyslipidemia, glucose intolerance and/or insulin resistance, and hypertension. Generally, MetS is accompanied by an exacerbation of oxidative stress, inflammation, and vascular dysfunction. Increasing evidence suggests that berries and berry bioactives could play a potential role in the prevention and mitigation of the risk factors associated with MetS. The present systematic review summarizes the more recently available evidence deriving from human intervention studies investigating the effect of berries in subjects with at least three out of five MetS parameters. The PubMed, Scopus, and Embase databases were systematically searched from January 2010 until December 2022. A total of 17 human intervention trials met the inclusion criteria. Most of them were focused on blueberry (n = 6), cranberry (n = 3), and chokeberry (n = 3), while very few or none were available for the other berries. If considering MetS features, the main positive effects were related to lipid profile (low and high-density lipoproteins, cholesterol, and triglycerides) following blueberries and chokeberries, while conflicting results were documented for anthropometric parameters, blood pressure, and fasting blood glucose levels. Other markers analyzed within the studies included vascular function, oxidative stress, and inflammation. Here, the main positive effects were related to inflammation with a reduction in interleukin 6 and tumor necrosis factor-alpha following the intake of different berries. In conclusion, although limited, the evidence seems to support a potential role for berries in the modulation of lipid profile and inflammation in subjects with MetS. Furthermore, high-quality intervention trials are mandatory to demonstrate the role of berries in reducing risk factors for MetS and related conditions. In the future, such a demonstration could bring the adoption of berries as a potential dietary strategy to prevent/counteract MetS and related risk factors.

Ready-To-Eat Rocket Salads as Potential Reservoir of Bacteria for the Human Microbiome.

Reportedly, Western-type diets may induce the loss of key microbial taxa within the gastrointestinal microbiota, promoting the onset of noncommunicable diseases. It was hypothesized that the consumption of raw vegetables could contribute to the maintenance of the intestinal microbial community structure. In this context, we explored bacteria associated with commercial rocket salads produced through different farming practices: traditional (conventional, organic, and integrated) and vertical farming. Viable counts of mesophilic bacteria and lactic acid bacteria (LAB) were performed on plate count agar (PCA) and de Man-Rogosa-Sharpe (MRS) agar at pH 5.7, whereas metataxonomics through 16S rRNA gene sequencing was used to profile total bacteria associated with rocket salads. We found that rocket salads from vertical farming had much fewer viable bacteria and had a bacterial community structure markedly different from that of rocket salads from traditional farming. Furthermore, although α- and ß-diversity analyses did not differentiate rocket samples according to farming techniques, several bacterial taxa distinguished organic and integrated from conventional farming salads, suggesting that farming practices could affect the taxonomic composition of rocket bacterial communities. LAB were isolated from only traditional farming samples and belonged to different species, which were variably distributed among samples and could be partly associated with farming practices. Finally, the INFOGEST protocol for in vitro simulation of gastrointestinal digestion revealed that several taxonomically different rocket-associated bacteria (particularly LAB) could survive gastrointestinal transit. This study suggests that commercial ready-to-eat rocket salads harbor live bacteria that possess the ability to survive gastrointestinal transit, potentially contributing to the taxonomic structure of the human gut microbiota. IMPORTANCE Western-type diets are composed of foods with a reduced amount of naturally occurring microorganisms. It was hypothesized that a microbe-depleted diet can favor the alteration of the human intestinal microbial ecosystem, therefore contributing to the onset of chronic metabolic and immune diseases currently recognized as the most significant causes of death in the developed world. Here, we studied the microorganisms that are associated with commercial ready-to-eat rocket salads produced through different farming practices. We showed that rocket salad (a widely consumed vegetal food frequently eaten raw) may be a source of lactic acid bacteria and other microbes that can survive gastrointestinal transit, potentially increasing the biodiversity of the intestinal microbiota. This deduction may be valid for virtually all vegetal foods that are consumed raw.

Blueberries and their bioactives in the modulation of oxidative stress, inflammation and cardio/vascular function markers: a systematic review of human intervention studies.

Blueberries represent a rich source of (poly)phenols and other bioactive compounds. Numerous in vitro and animal model studies documented the potential health-promoting properties of blueberries and blueberry-bioactives, while little is still known about their effects in humans. The objective of the present systematic review is to provide main evidence and the potential mechanisms of action of blueberry and its (poly)phenols in the regulation of markers related to oxidative stress, inflammation, vascular and cardiometabolic function in health and disease states. A total of 45 human intervention studies were included in this review. Overall, the evidence suggests that blueberries may play a role in the improvement of markers of vascular function. Their effects were observed following both post-prandial and long-term consumption, particularly in subjects with risk factors and/or disease conditions. Conversely, the conflicting results on inflammation, oxidative stress and cardiometabolic risk markers were most likely due to differences among studies in terms of study design, subject characteristics, duration of intervention, dosage, and type of biomarkers analyzed. For these reasons, high-quality, well-designed, human intervention studies are warranted to strengthen the current findings on vascular function and provide more evidence about the impact of blueberries on the different markers considered. In addition, studies focusing on the relationship between the structure and the function of (poly)phenols will be fundamental for a better comprehension of the mechanisms behind the health effects observed.

Acute effect of blueberry intake on vascular function in older subjects: Study protocol for a randomized, controlled, crossover trial.

Aging is associated with an increased risk of developing cardiovascular disease which is often accompanied by a decline in vascular health and function. Current evidence suggests that berries may have a potential role in the modulation of vascular function, but dietary interventions are still needed to confirm findings, especially in older subjects. In the context of the MIND FoodS HUB project, this study aims to investigate the effect of a single serving of blueberry (250 g of blueberry versus a control product) in a group of older subjects (≥ 60y) through a randomized, controlled, cross-over dietary intervention trial. Specifically, the study evaluates the absorption kinetics of bioactives following the blueberries intake and the effects on markers related to oxidative stress, inflammation, and vascular function analyzed at different time points. By considering a drop-out rate estimate of 25%, at least 20 subjects will be recruited in the study. The study will provide evidence to support the potential beneficial effects of blueberry and its bioactive compounds on vascular function in a group of population more susceptible to vascular dysfunction and to the development of cardiovascular diseases. Moreover, the study will contribute the analysis of several metabolic and functional markers that can support the biological plausibility of the results obtained. Finally, the trial will provide data on the absorption and metabolism of blueberry bioactives which will be used to study their association with the different markers under study. Trail registration: The trial is registered at ISRCTN (http://isrctn.com/ISRCTN18262533); May 7, 2021.

Flavonoid Intake in Relation to Colorectal Cancer Risk and Blood Bacterial DNA.

Flavonoids have been inversely associated to colorectal cancer (CRC) and are plausible intermediaries for the relation among gut microbiome, intestinal permeability and CRC. We analyzed the relation of flavonoid intake with CRC and blood bacterial DNA. We conducted a case-control study in Italy involving 100 incident CRC cases and 200 controls. A valid and reproducible food-frequency questionnaire was used to assess dietary habits and to estimate six flavonoid subclass intakes. We applied qPCR and 16S rRNA gene profiling to assess blood bacterial DNA. We used multiple logistic regression to derive odds ratios (ORs) of CRC and Mann-Whitney and chi--square tests to evaluate abundance and prevalence of operational taxonomic units (OTUs) according to flavonoid intakes. Inverse associations with CRC were found for anthocyanidins (OR for the highest versus the lowest tertile = 0.24, 95% confidence interval, CI = 0.11-0.52) and flavanones (OR = 0.18, 95% CI = 0.08-0.42). We found different abundance and prevalence according to anthocyanidin and flavanone intake for OTUs referring to Oligoflexales order, Diplorickettsiaceae family, Staphylococcus, Brevundimonas, Pelomonas and Escherischia-Shigella genera, and Flavobacterium and Legionella species. The study provides evidence to a protective effect of dietary anthocyanidins and flavanones on CRC and suggests an influence of flavonoids on blood bacterial DNA, possibly through intestinal permeability changes.