Inhibition of CpLIP2 Lipase Hydrolytic Activity by Four Flavonols (Galangin, Kaempferol, Quercetin, Myricetin) Compared to Orlistat and Their Binding Mechanisms Studied by Quenching of Fluorescence.

The inhibition of recombinant CpLIP2 lipase/acyltransferase from Candida parapsiolosis was considered a key model for novel antifungal drug discovery and a potential therapeutic target for candidiasis. Lipases have identified recently as potent virulence factors in C. parapsilosis and some other yeasts. The inhibition effects of orlistat and four flavonols (galangin, kaempferol, quercetin and myricetin) characterized by an increasing degree of hydroxylation in B-ring, were investigated using ethyl oleate hydrolysis as the model reaction. Orlistat and kaempferol (14 µM) strongly inhibited CpLIP2 catalytic activity within 1 min of pre-incubation, by 90% and 80%, respectively. The relative potency of flavonols as inhibitors was: kaempferol > quercetin > myricetin > galangin. The results suggested that orlistat bound to the catalytic site while kaempferol interacted with W294 on the protein lid. A static mechanism of interactions between flavonols and CpLIP2 lipase was confirmed by fluorescence quenching analyses, indicating that the interactions were mainly driven by hydrophobic bonds and electrostatic forces. From the Lehrer equation, fractions of tryptophan accessibility to the quencher were evaluated, and a relationship with the calculated number of binding sites was suggested.

Validation of a surgical technique for rat intestinal irradiation: potential side effects prevention by dietary grape phenolics.

AIMS: This study evaluates and defines the histological and biochemical consequences of irradiation on the Hauer-Jensen intestinal model and investigates the potential effects of dietary polyphenols. MAIN METHODS: Sprague-Dawley rats were orchiectomized, and an ileal loop was transposed to the left part of the scrotum, then irradiated 2 weeks after surgery with a single dose of 21 Gy (4.49 Gy/min). Four groups of rats received either phenolic extracts from grape seeds (EGS) and from red wine (ACYS, EGT), or pure quercetin 3-O-ß-glucoside (Q3G), for 5 days before the irradiation and were sacrificed 2 weeks after. Antioxidant enzyme activities, i.e. superoxide dismutase (SOD) and glutathione peroxidase activity (GSHPx), and oxidative markers such as myeloperoxidase activity (MPO) and thiobarbituric acid reactive substances (MDA) were measured as well as cytokine-induced neutrophil chemoattractant level (CINC-1), a chemokine involved in inflammation. KEY FINDINGS: Irradiated rats exhibited a high radiation injury score (RIS) with a thickened serosa, mucosal loss and ulceration, and epithelial atypicality. Intestinal MPO activity and CINC-1 concentration were significantly increased in irradiated animals (60 and 66 %, respectively). Higher plasma MDA levels (58 %) and SOD activity (32 %) were accompanied by a reduced GSHPx activity (79 %). However, feeding phenolic extracts remarkably reduced levels of blood SOD activity (34 % on average), intestinal CINC-1 (25-75 % range) and MPO activity (36-84 %). Except for Q3G, phenolics preserved the intestinal structure. SIGNIFICANCE: These findings show that irradiation triggers an inflammation, and an oxidative stress by disturbing the pro-oxidant/antioxidant balance and indicate that phenolics supply exerts preventive effects against radio-induced intestinal impairment.

Structure of PlcR: Insights into virulence regulation and evolution of quorum sensing in Gram-positive bacteria.

Gram-positive bacteria use a wealth of extracellular signaling peptides, so-called autoinducers, to regulate gene expression according to population densities. These "quorum sensing" systems control vital processes such as virulence, sporulation, and gene transfer. Using x-ray analysis, we determined the structure of PlcR, the major virulence regulator of the Bacillus cereus group, and obtained mechanistic insights into the effects of autoinducer binding. Our structural and phylogenetic analysis further suggests that all of those quorum sensors that bind directly to their autoinducer peptide derive from a common ancestor and form a single family (the RNPP family, for Rap/NprR/PlcR/PrgX) with conserved features. As a consequence, fundamentally different processes in different bacterial genera appear regulated by essentially the same autoinducer recognition mechanism. Our results shed light on virulence control by PlcR and elucidate origin and evolution of multicellular behavior in bacteria.

Influence of packaging conditions on natural microbial population growth of endive.

The influence of three packaging conditions, i.e., unmodified atmosphere packaging (UAP), passive modified atmosphere packaging (MAP), and active MAP, on the natural microbial population growth of endive was investigated at 20 degrees C. For UAP, endive was placed in macroperforated oriented polypropylene pouches that maintained gas composition close to that of air (21 kPa O2 and 0 kPa CO2) but also limited superficial product dehydration. For MAP, endive was placed in low-density polyethylene pouches that induced a 3 kPa O2 and 5 kPa CO2 equilibrium atmosphere composition. Steady state was reached after 25 h of storage with an oxygen absorbing packet (active MAP) compared with 100 h without the packet (passive MAP) and was maintained for 200 h. After 312 h of storage, both active and passive MAP reduced total aerobic mesophile, yeast, and mold population growth compared with endive in UAP. Active MAP accelerated and improved the inhibition of Pseudomonas spp. and Enterobacteriaceae, respectively, probably because of the rapid O2 depletion during the transition period. A shift in the Enterobacteriaceae subpopulation from Rhanella aquatilis to Enterobacter agglomerans was observed for both passive and active MAP.

Moderate chronic administration of Vineatrol-enriched red wines improves metabolic, oxidative, and inflammatory markers in hamsters fed a high-fat diet.

SCOPE: High-fat (HF) diets contribute to the development of cardiovascular diseases and the metabolic syndrome. This study was undertaken to investigate the beneficial effects of Vineatrol®-enriched red wines on blood lipids, oxidative stress and inflammation, and the role of some metabolic pathway regulatory proteins. METHODS AND RESULTS: Golden Syrian hamsters received an HF diet for 13 wk, in the presence or absence of red wines supplemented with Vineatrol® (RWV) or not. The HF diet increased plasma cholesterol, triglycerides, glucose, and insulin, which were attenuated by RWV treatment. RWV protected against the HF-induced increase in liver nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity and spared antioxidant enzyme activities. RWV did not reduce either liver steatosis or increased plasma leptin due to the HF diet, but greatly improved adiponectinemia. In the liver, RWV affected the inflammatory response by decreasing polymorphonuclear cell number and lowering TNF-α and IL-6 levels. Moreover, the increase in NF-κB activity in the HF group liver was prevented by RWV. Finally, RWV partially corrected low SIRT1 levels due to the HF diet but had no influence on SIRT3 or p-AMPK protein levels. CONCLUSION: Our studies suggest that RWV is capable of reversing the atherogenic process induced by an HF diet in hamster tissues.