Jacob Lessard-Lord

Inter-individual variability in gVLM _B microbial metabolism and the short-excretion kinetics of SREM _B limit biomarkers reclassification of flavanol intake in the COSMOS trial, warranting dedicated validation for intended use.

Heterozygous familial hypercholesterolemia (HeFH) is a genetic disorder that accelerates atherosclerosis and leads to premature cardiovascular diseases (CVD). Whole food, plant-based diets (WFPB) are recommended worldwide for their cardioprotective properties but evidence regarding their effects in HeFH management remains unavailable. This study aims to evaluate the impact of a WFPB, in place of a standard American diet (SAD), on LDL-cholesterol (LDL-C) (primary outcome) and other CVD risk factors among adults with HeFH. In this randomized, two-period, two-treatment, crossover, controlled feeding trial, 50 adults with genetically confirmed HeFH, free of cholesterol-lowering medication, consumed a WFPB and a SAD for 4 weeks each in a random order, under fully controlled, isocaloric feeding conditions. The diets were separated by a two- to four-week washout period. LDL-C and other CVD risk factors were measured at the end of each diet. The WFPB induced a clinically significant reduction in LDL-C relative to the SAD ( − 17.9%, 95% CI: −21.7%, −14.3%; P < 0.0001). The study demonstrates the clinical significance of diet therapy in HeFH and supports its re-establishment as a cornerstone in clinical guidelines (clinicaltrials.gov registration: NCT05181553; funding: Canadian Institutes of Health Research).

Flavan-3-ols intake is associated with numerous health benefits, but these are influenced by their conversion into smaller phenolic metabolites by the gut microbiota. Thus, the identification of bacteria that metabolize flavan-3-ols could lead to targeted interventions to enhance their benefits. To this end, we screened 47 Lactiplantibacillus plantarum strains for their ability to metabolize (+)-catechin, a flavan-3-ol. Then, we assessed these strains for their capacity to convert various flavan-3-ol structures. Out of the 47 isolates, 12 released 3-(3',4'-dihydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)-propan-2-ol (a form of diphenylpropan-2-ol) from (+)-catechin. All strains metabolized (+)-catechin, (-)-epicatechin, (-)-epigallocatechin, but only a subset transformed (-)-gallocatechin. Among these simple flavan-3-ol structures, (-)-epicatechin was metabolized the most. A hierarchical cluster analysis identified two groups of flavan-3-ol-metabolizing strains categorized as having "high" and "low" production of diphenylpropan-2-ols. Notably, the strains that produced higher levels of diphenylpropan-2-ol from (+)-gallocatechin and (+)-catechin also performed better with a camu-camu extract, which was studied as a complex source of flavan-3-ols and predominantly contained these two flavan-3-ols. These results demonstrate the interstrain variability in L. plantarum metabolism, which may be useful for developing tailored formulations to enhance the production of flavan-3-ols bioactive metabolites.

Some Lactiplantibacillus plantarum strains produce an extracellular tannase that converts galloylated ellagitannins into ellagitannin-core structures and bioactive phenolic metabolites, thereby promoting the health effects of these polyphenols.

Background: Proanthocyanidins (PAC) and oligosaccharides from cranberry exhibit multiple bioactive health properties and persist intact in the colon post-ingestion. They display a complex bidirectional interaction with the microbiome, which varies based on both time and specific regions of the gut; the nature of this interaction remains inadequately understood. Therefore, we aimed to investigate the impact of cranberry extract on gut microbiota ecology and function. Methods: We studied the effect of a cranberry extract on six healthy participants over a two-week supplementation period using the ex vivo artificial fermentation system TWIN-M-SHIME to replicate luminal and mucosal niches of the ascending and transverse colon. Results: Our findings revealed a significant influence of cranberry extract supplementation on the gut microbiota ecology under ex vivo conditions, leading to a considerable change in bacterial metabolism. Specifically, Bifidobacterium adolescentis (B. adolescentis ) flourished in the mucus of the ascending colon, accompanied by a reduced adhesion of Proteobacteria . The overall bacterial metabolism shifted from acetate to propionate and, notably, butyrate production following PAC supplementation. Although there were variations in microbiota modulation among the six donors, the butyrogenic effect induced by the supplementation remained consistent across all individuals. This metabolic shift was associated with a rise in the relative abundance of several short-chain fatty acid (SCFA)-producing bacterial genera and the formation of a consortium of key butyrogenic bacteria in the mucus of the transverse colon. Conclusions: These observations suggest that cranberry extract supplementation has the potential to modulate the gut microbiota in a manner that may promote overall gut health.

Cranberry is associated with multiple health benefits, which are mostly attributed to its high content of (poly)phenols, particularly flavan-3-ols. However, clinical trials attempting to demonstrate these positive effects have yielded heterogeneous results, partly due to the high inter-individual variability associated with gut microbiota interaction with these molecules. In fact, several studies have demonstrated the ability of these molecules to modulate the gut microbiota in animal and in vitro models, but there is a scarcity of information in human subjects. In addition, it has been recently reported that cranberry also contains high concentrations of oligosaccharides, which could contribute to its bioactivity. Hence, the aim of this study was to fully characterize the (poly)phenolic and oligosaccharidic contents of a commercially available cranberry extract and evaluate its capacity to positively modulate the gut microbiota of 28 human subjects. After only four days, the (poly)phenols and oligosaccharides-rich cranberry extract, induced a strong bifidogenic effect, along with an increase in the abundance of several butyrate-producing bacteria, such as Clostridium and Anaerobutyricum. Plasmatic and fecal short-chain fatty acids profiles were also altered by the cranberry extract with a decrease in acetate ratio and an increase in butyrate ratio. Finally, to characterize the inter-individual variability, we stratified the participants according to the alterations observed in the fecal microbiota following supplementation. Interestingly, individuals having a microbiota characterized by the presence of Prevotella benefited from an increase in Faecalibacterium with the cranberry extract supplementation.

Clinical trials investigating the health effects of flavan‐3‐ols yield heterogeneous results due to interindividual variability in the gut microbiota metabolism. In fact, different groups in the population have similar metabolic profiles following (−)‐epicatechin and (+)‐catechin gut microbial metabolism and can be regrouped into so‐called metabotypes. In this study, the capacity of 34 donors to metabolize polymeric B‐type flavan‐3‐ols from aronia and oligomeric A‐type flavan‐3‐ols from cranberry is investigated by in vitro fecal batch fermentations. Less than 1% of the flavan‐3‐ols from both sources are converted into microbial metabolites, such as phenyl‐γ‐valerolactones (PVLs). To further confirm this result, gut microbial metabolites from flavan‐3‐ols are quantified in urine samples collected from participants, before and after a 4‐day supplementation of cranberry extract providing 82.3 mg of flavan‐3‐ols per day. No significant difference is observed in the urinary excretion of flavan‐3‐ols microbial metabolites. Hence, it demonstrates by both in vitro and in vivo approaches that flavan‐3‐ols from aronia and cranberry are poorly degraded by the gut microbiota. The beneficial health impacts of these molecules likely stem from their capacity to affect gut microbiota and their interactions with the gut epithelium, rather than from their breakdown into smaller metabolites.

In vitro systems are widely employed to assess the impact of dietary compounds on the gut microbiota and their conversion into beneficial bacterial metabolites. However, the complex fluid dynamics and multi‐segmented nature of these systems can complicate the comprehensive analysis of dietary compound fate, potentially confounding physical dilution or washout with microbial catabolism. In this study, we developed fluid dynamics models based on sets of ordinary differential equations to simulate the behavior of an inert compound within two commonly used in vitro systems: the continuous two‐stage PolyFermS system and the semi‐continuous multi‐segmented SHIME® system as well as into various declinations of those systems. The models were validated by investigating the fate of blue dextran, demonstrating excellent agreement between experimental and modeling data (with r ² values ranging from 0.996 to 0.86 for different approaches). As a proof of concept for the utility of fluid dynamics models in in vitro system, we applied generated models to interpret metabolomic data of procyanidin A2 (ProA2) generated from the addition of proanthocyanidin (PAC)‐rich cranberry extract to both the PolyFermS and SHIME® systems. The results suggested ProA2 degradation by the gut microbiota when compared to the modeling of an inert compound. Models of fluid dynamics developed in this study provide a foundation for comprehensive analysis of gut metabolic data in commonly utilized in vitro PolyFermS and SHIME® bioreactor systems and can enable a more accurate understanding of the contribution of bacterial metabolism to the variability in the concentration of target metabolites.

Quantification of nutritional biomarkers is crucial to accurately assess the dietary intake of different classes of (poly)phenols in large epidemiological studies. High-throughput analysis is mandatory to apply this methodology in large cohorts. However, the current validated methods to quantify (poly)phenols metabolites in biological fluids use ultra performance liquid chromatography (UPLC), leading to analysis time of several minutes per sample. To significantly reduce the run time, we developed and validated a method to quantify in urine the flavan-3-ols biomarkers, phenyl-gamma-valerolactones (PVLs), using laser diode thermal desorption (LDTD). This mass spectrometry source allows direct introduction of sample extracts, resulting in analysis time of less than 10 s per sample. Also, to encompass the problem associated with the cost and availability of sulfated and glucuronide analytical standards, urine samples were subjected to enzymatic hydrolysis. Creatinine was also quantified to normalize the results obtained from the urinary spot. Results obtained with LDTD-MS/MS were cross-validated by UPLC-MS/MS using 155 urine samples. Coefficient of correlation was above 0.975 for PVLs and creatinine. For all analytes, the accuracy was between 90% and 113% by LDTD-MS/MS. Altogether, sample preparation was fully automated to demonstrate the application potential of this method to large cohorts.

Although the relationship between gut microbiota and flavan-3-ol metabolism differs greatly between individuals, the specific metabolic profiles, known as metabotypes, have not yet been clearly defined. In this study, fecal batch fermentations of 34 healthy donors inoculated with (-)-epicatechin were stratified into groups based on their conversion rate of (-)-epicatechin and their quali-quantitative metabolic profile. Fast and slow converters of (-)-epicatechin, high producers of 1-(3'-hydroxyphenyl)-3-(2″,4″,6″-trihydroxyphenyl)-propan-2-ol (3-HPP-2-ol) and 5-(3',4'-dihydroxyphenyl)-γ-valerolactone (3,4-DHPVL) were identified. Fecal microbiota analysis revealed that fast conversion of (-)-epicatechin was associated with short-chain fatty acid (SCFA)-producing bacteria, such as Faecalibacterium spp. and Bacteroides spp., and higher levels of acetate, propionate, butyrate, and valerate were observed for fast converters. Other bacteria were associated with the conversion of 1-(3',4'-dihydroxyphenyl)-3-(2″,4″,6″-trihydroxyphenyl)-propan-2-ol into 3-HPP-2-ol (Lachnospiraceae UCG-010 spp.) and 3,4-DHPVL (Adlercreutzia equolifaciens). Such stratification sheds light on the mechanisms of action underlying the high interindividual variability associated with the health benefits of flavan-3-ols.