Rational design, synthesis, and evaluation of nanomolar type II dehydroquinase inhibitors

The in silico design, synthesis, and biological evaluation of ten potent type II dehydroquinase inhibitors are described. These compounds contain an anhydroquinate core, incorporated as a mimic of the enolate reaction intermediate. This substructure is attached by a variety of linking units to a terminal phenyl group that binds in an adjacent pocket. Inhibitors were synthesised from (-)-quinic acid using palladium-catalysed Stille and carboamidation chemistry. Several inhibitors exhibited nanomolar inhibition constants against type II dehydroquinases from Streptomyces coelicolor and Mycobacterium tuberculosis. These are among the most potent inhibitors of these enzymes reported to date.     

Tracking a new cell-penetrating (W/R) nonapeptide, through an enzyme-stable mass spectrometry reporter tag

We have designed a mass stable reporter (msr) tag with m/z over 500, trifluoroacetyl(alpha,alpha-diethyl)Gly-Lys(Nepsilonbiotin)-(D)Lys-Cys, for the quantification of the uptake and study of the degradation processes of cell-penetrating peptides (CPP), by matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. This tag was found stable in cell lysis conditions. Using a quantitative MALDI-TOF mass spectrometry analysis based method, an accurate tracking of a new CPP and of its degradation products could be done. (1) The new msr(W/R) nonapeptide (H-RRWWRRWRR-NH2) enters chinese hamster ovary (CHO) K1 cells with a kinetic reaching a steady state after 30-60 min of incubation. This plateau was stable for 4 h and decreased slowly afterward. (2) The peptide msr(W/R) nonapeptide was not cytotoxic over 48 h incubation with CHO cells. (3) After 1 h incubation, the msr(W/R) nonapeptide accumulated with a 3-fold higher concentration than the extracellularly added concentration (7.5 microM). (4) The intracellular quantification was accurate with less than 3% of the quantified peptide being potentially membrane-bound. (5) There was no leakage of the full-length CPP outside the cells. And, finally, (6) analysis of the degradation process of this new CPP suggests that the peptide did not traffick to lysosomes.     

Fish Oil and Microalga Omega-3 as Dietary Supplements: A Comparative Study on Cardiovascular Risk Factors in High-Fat Fed Rats

Dietary supplementation with marine omega‐3 polyunsaturated fatty acids (n‐3 PUFA) can have beneficial effects on a number of risk factors for cardiovascular disease (CVD). We compared the effects of two n‐3 PUFA rich food supplements (freeze‐dried Odontella aurita and fish oil) on risk factors for CVD. Male rats were randomly divided into four groups of six animals each and fed with the following diets: control group (C) received a standard diet containing 7 % lipids; second group (HF high fat) was fed with a high‐fat diet containing 40 % lipids; third group (HFFO high fat+fish oil) was fed with the high‐fat diet supplemented with 0.5 % fish oil; and fourth group (HFOA high fat+O. aurita) received the high‐fat diet supplemented with 12 % of freeze‐dried O. aurita. After 8 weeks rats fed with the high‐fat diet supplemented with O. aurita displayed a significantly lower bodyweight than those in the other groups. Both the microalga and the fish oil significantly reduced insulinemia and serum lipid levels. O. aurita was more effective than the fish oil in reducing hepatic triacyglycerol levels and in preventing high‐fat diet‐induced steatosis. O. aurita and fish oil also reduced platelet aggregation and oxidative status induced by high fat intake. After an OA supplementation, the adipocytes in the HFOA group were smaller than those in the HF group. Freeze‐dried O. aurita showed similar or even greater biological effects than the fish oil. This could be explained by a potential effect of the n‐3 PUFA but also other bioactive compounds of the microalgae.     

Site‐Specific Labeling of MicroRNA Precursors: A Structure–Activity Relationship Study

Functionalized oligoribonucleotides are essential tools in RNA chemical biology. Various synthetic routes have been developed over recent years to conjugate functional groups to oligoribonucleotides. However, the presence of the functional group on the oligoribonucleotide backbone can lead to partial or total loss of biological function. The limited knowledge concerning the positioning of functional groups therefore represents a hurdle for the development of oligoribonucleotide chemical tools. Here we describe a systematic investigation of site‐specific labeling of pre‐miRNAs to identify positions for the incorporation of functional groups, in order not to hinder their processing into active mature miRNAs.     

Syntheses,characterization, and hydrolytic degradation of P(ε‐caprolactone‐co‐δ‐valerolactone) copolymers: Influence of molecular weight

Biodegradable poly(ε‐caprolactone‐co‐δ‐valerolactone) copolymers were synthesized and investigated to study their behavior in aqueous medium. The copolyesters were produced by ring opening polymerization between ε‐caprolactone (CL) and δ‐valerolactone (VL) in bulk at 140°C using tin(II) octoate as catalyst. They were characterized by using ¹H NMR, size exclusion chromatography, differential scanning calorimetry, and MALDI TOF mass spectrometry. Reactivity ratio determination gave an insight on their microstructure. Hydration, hydrolytic degradation, and biocide release of P(CL‐VL) films with different molecular weights values were studied. A one‐order kinetic whose rate constant decreases with copolymer macromolecular weight was observed. Although the molecular weight decrease remained relatively weak after 8 months of immersion, a correlation between molecular weight and hydrolysis rate was shown by high performance liquid chromatography‐mass spectrometry. The ability of the P(CL‐VL) films to release active compounds dispersed in the films was studied by atomic absorption spectroscopy. The release behavior of all copolymers was identical with a zero‐order kinetic. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43007.     

Oxygen atom density in capacitively coupled RF oxygen plasma

The density of neutral oxygen atoms was determined in a plasma reactor for surface functionalization of polymer materials. Plasma was created in a stainless steel chamber by capacitively coupled RF generator at 13.56 MHz and adjustable forward power up to 100 W. Measurements were performed with a classical nickel catalytic probe. Systematic measurements were performed at a constant pumping speed, different flow rates from 15 to 100 sccm corresponding to pressures between 30 and 110 Pa, different powers between 40 and 100 W and different probe positions in the discharge chamber. The results showed that the O atom density did not depend much on probe position as long as it was between the powered electrode and grounded housing facing the electrode. The O density depended rather linearly with power at fixed pressure. At low power, the O density did not depend much on pressure, but at high power, it was increasing with increasing pressure. The O density was of the order of 10¹⁹ m⁻³ and increased slightly over 10²⁰ m⁻³ at the highest power and pressure. The results were explained by gas phase and surface reactions.     

Supplementation of pig diet with algal fibre changes the chemical and physicochemical characteristics of digesta

Seaweed extracts, because of their physicochemical characteristics and potential nutritional value, could provide a new source of dietary fibre. This study investigated changes in seaweed fibres (physicochemical and fermentative properties) in different digestive sites and their effects on digesta (viscosity and hydration properties). Sixteen pigs were adapted to a test diet supplemented with 5% algal fibre (either Palmaria palmata (PP), a poorly viscous soluble xylan; or Eucheuma cottonii (EC), a partly insoluble carrageenan; or Laminaria digitata (LD), a highly viscous soluble alginate) or 5% cellulose (reference fibre). PP did not modify the characteristics of digesta and was fermented in the caecum (pH 6.1 ± 0.4; short‐chain fatty acids measured in digesta, 1409 ± 691 µmol g⁻¹ dry matter). EC and LD were mainly insoluble in the stomach, becoming soluble in the intestine; EC was slightly fermented in the colon, giving a low concentration of short‐chain fatty acids (303 ± 122 µmol g⁻¹ dry matter). Supplementation of the diet with alginate (LD) increased 3.5‐fold the ileal viscosity of digesta and their hydration capacity in the ileum and colon. Thus the physicochemical properties of pig digesta largely depend on the physicochemical properties of the ingested seaweed fibre, the pH and ionic conditions prevailing in the gut, and their fermentability. © 2000 Society of Chemical Industry     

Characterization of the "viable but nonculturable" (VBNC) state in the wine spoilage yeast Brettanomyces

Although the viable but not culturable (VBNC) state has been studied in detail in bacteria, it has been suggested that maintenance of viability with loss of culturability also exists in eukaryotic cells, such as in the wine spoilage yeast Brettanomyces. To provide conclusive evidence for the existence of a VBNC state in this yeast, we investigated its capacity to become viable and nonculturable after sulfite stress, and its ability to recover culturability after stressor removal. Sulfite addition induced loss of culturability but maintenance of viability. Increasing the medium pH to decrease the concentration of toxic SO(2) allowed yeast cells to become culturable again, thus demonstrating the occurrence of a VBNC state in Brettanomyces upon SO(2) exposure. Relative to culturable Brettanomyces, VBNC yeast cells were found to display a 22% decrease in size on the basis of laser granulometry. Assays for 4-ethylguaiacol and 4-ethylphenol, volatile phenols produced by Brettanomyces, indicated that spoilage compound production could persist in VBNC cells. These morphological and physiological changes in VBNC Brettanomyces were coupled to extensive protein pattern modifications, as inferred by comparative two-dimensional electrophoresis and mass spectrometric analyses. Upon identification of 53 proteins out of the 168 spots whose abundance was significantly modified in treated cells relative to control, we propose that the SO(2)-induced VBNC state in Brettanomyces is characterized by a reduced glycolytic flux coupled to changes in redox homeostatis/protein turnover-related processes. This study points out the existence of common mechanisms between yeast and bacteria upon entry to the VBNC state.