Scope
Methods to verify cranberry juice consumption are lacking. Predictive multivariate models built upon validated biomarkers may help to verify human consumption of a food using a nutrimetabolomics approach.
Methods
A 21‐day double‐blinded, randomized, placebo‐controlled, cross‐over study was conducted among healthy young women aged 1829. Plasma was collected at baseline and after 3‐day and 21‐day consumption of cranberry or placebo juice. Plasma metabolome was analyzed using UHPLC coupled with high resolution mass spectrometry.
Results
18 discriminant metabolites in positive mode and 18 discriminant metabolites in negative mode from a previous 3‐day open‐label study were re‐discovered in the present blinded study. Predictive orthogonal partial least squares discriminant analysis (OPLS‐DA) models were able to identify cranberry juice consumers over a placebo juice group with 96.9% correction rates after 3‐day consumption in both positive and negative mode. This present study revealed 84 and 109 additional discriminant metabolites in positive and negative mode, respectively. Twelve of them were tentatively identified.
Conclusion
Cranberry juice consumers were classified with high correction rates using predictive OPLS‐DA models built upon validated plasma biomarkers. Additional biomarkers were tentatively identified. These OPLS‐DA models and biomarkers provided an objective approach to verify participant compliance in future clinical trials.
This study examined the ability of five Amberlite resins coupled with ultrasound-assisted water extraction for the recovery and enrichment of bioactive procyanidins and total phenolics from cranberry pomace. Static adsorption showed that XAD-7HP had the highest adsorption capacity for procyanidins (52.2 mg/g resin) and total phenolics (99.1 mg/g resin) whereas XAD-761 had the lowest. Adsorption of procyanidins fitted better to pseudo-second-order kinetics than pseudo-first-order kinetics. Isotherm adsorption on XAD-7HP suggested that Langmuir isotherm was a better model to describe the adsorption of procyanidins while Kemkin-Pyzhev equation was better for total phenolics based on higher coefficient of determinations (R ). Dynamic tests on XAD-7HP suggested that the flow rate of 7 and 8 mL/min were the optimum conditions for adsorption and desorption of procyanidins, respectively. Measurements using HPLC revealed that adsorption increased the contents of procyanidins and total phenolics by 4.57- and 4.73-folds, respectively, compared to the initial extracts. This research showed that Amberlite XAD-7HP resin adsorption coupled with ultrasound-assisted water extraction is an efficient method to separate and concentrate procyanidins from cranberry pomace.
Despite considerable advances in surgical repairing procedures, congenital heart diseases (CHDs) remain the leading noninfectious cause of infant morbidity and mortality. Understanding the molecular/genetic mechanisms underlying normal cardiogenesis will provide essential information for the development of novel diagnostic and therapeutic strategies against CHDs. BMP signaling plays complex roles in multiple cardiogenic processes in mammals. SMAD1 is a canonical nuclear mediator of BMP signaling, the activity of which is critically regulated through its interaction partners. We screened a mouse embryonic heart yeast two-hybrid library using Smad1 as bait and identified SERTAD1 as a novel interaction partner of SMAD1. SERTAD1 contains multiple potential functional domains, including two partially overlapping transactivation domains at the C terminus. The SERTAD1-SMAD1 interaction in vitro and in mammalian cells was further confirmed through biochemical assays. The expression of Sertad1 in developing hearts was demonstrated using RT-PCR, western blotting and in situ hybridization analyses. We also showed that SERTAD1 was localized in both the cytoplasm and nucleus of immortalized cardiomyocytes and primary embryonic cardiomyocyte cultures. The overexpression of SERTAD1 in cardiomyocytes not only enhanced the activity of two BMP reporters in a dose-dependent manner but also increased the expression of several known BMP/SMAD regulatory targets. Therefore, these data suggest that SERTAD1 acts as a SMAD1 transcriptional co-activator to promote the expression of BMP target genes during mouse cardiogenesis.
Objectives
Adhesion of type-P and type-1 fimbriated uropathogenic E. coli to urinary tract epithelial cells initiates urinary tract infections. This research aimed to optimize and apply a fluorometric method to evaluate the capacities of cranberry polyphenols and metabolites to inhibit such adhesion in vitro.
Methods
BacLight Green labelled E. coli were incubated with cranberry polyphenols or microbial metabolites of cranberry polyphenols for 30 min at 37°C. Mixture was added to a 96-well microplate containing 1 × 105/well of human uroepithelial T24 cells and incubated for 1 h at 37°C. After incubation, E. coli not adhered were removed by phosphate buffer washing. Fluorescent intensity was measured on a microplate reader at 480 nm excitation and 516 nm emission.
Results
Stable and strong fluorescent readings were obtained with 800 μmol/L BacLight Green for E. coli labeling and an E. coli to T24 cells ratio of 400:1 for co-incubation. A standard curve was established
using 0–63 μM myricetin. The half-maximal inhibitory concentrations (IC50) of myricetin were 13.2 μM against type-P E. coli adhesion and 5.5 μM against type-1 E. coli adhesion. A fraction enriched with procyanidin polymers had IC50 of 57.6 μg/mL against type-P E. coli and 19.3 μg/mL against type-1 E. coli, respectively. Its anti-adhesion activities were more potent than those of cranberry fractions enriched with procyanidin oligomers, flavonols, or anthocyanin. Procyanidin A2 had a maximal inhibition about 35% at 17.3 μM against type-P E. coli, but no anti-adhesion activity was observed against type-1 E. coli. Procyanidin B2 showed a plateaued inhibition about 15% at 173–691 μM against type-P E. coli. Its maximal inhibition against type-1 E. coli was around 25% at 346 μM. Hippuric acid, a major metabolite of cranberry polyphenols, had a maximal inhibition about 20% at 558 μM against type-1 E. coli adhesion, whereas its anti-adhesion activity against type-P E. coli was not detected.
Conclusions
The optimized fluorometric method showed that both structure and composition of cranberry polyphenols and metabolites affected their abilities to inhibit E. coli adhesion in vitro. Anti-adhesion activities of cranberry polyphenols also depend on type of E. coli fimbriae.
Funding Sources
University of Florida Research Foundation Seed Fund.
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