Molecular Networking for Drug Toxicities Studies: The Case of Hydroxychloroquine in COVID-19 Patients

Using drugs to treat COVID-19 symptoms may induce adverse effects and modify patient outcomes. These adverse events may be further aggravated in obese patients, who often present different illnesses such as metabolic-associated fatty liver disease. In Rennes University Hospital, several drug such as hydroxychloroquine (HCQ) have been used in the clinical trial HARMONICOV to treat COVID-19 patients, including obese patients. The aim of this study is to determine whether HCQ metabolism and hepatotoxicity are worsened in obese patients using an in vivo/in vitro approach. Liquid chromatography high resolution mass spectrometry in combination with untargeted screening and molecular networking were employed to study drug metabolism in vivo (patient’s plasma) and in vitro (HepaRG cells and RPTEC cells). In addition, HepaRG cells model were used to reproduce pathophysiological features of obese patient metabolism, i.e., in the condition of hepatic steatosis. The metabolic signature of HCQ was modified in HepaRG cells cultured under a steatosis condition and a new metabolite was detected (carboxychloroquine). The RPTEC model was found to produce only one metabolite. A higher cytotoxicity of HCQ was observed in HepaRG cells exposed to exogenous fatty acids, while neutral lipid accumulation (steatosis) was further enhanced in these cells. These in vitro data were compared with the biological parameters of 17 COVID-19 patients treated with HCQ included in the HARMONICOV cohort. Overall, our data suggest that steatosis may be a risk factor for altered drug metabolism and possibly toxicity of HCQ.     

Exploring natural products for new taste sensations

This paper discusses the discovery of uncommon taste or trigeminal active compounds and their associated sensory analysis using human tasting panels with the aim of enhancing the overall taste experience whilst reducing where possible the sugar and salt content of foods. The first example outlines the discovery of the sensory quality attributes of (R)-2-(carboxymethylamino)propanoic acid, named (R)-strombine, as assessed by a panel of 47 subjects to confirm its contribution to the typical taste of scallop muscle. The second example discusses the pungency and trigeminal effect of polygodial, which is compared with piperine and capsaicin, as well as the elucidation of a new structure eliciting a trigeminal effect, (+/-)-trans-2,3,3a,7a-tetrahydro-1 H-indene-4-carbaldehyde, discovered in Amomum tsao-ko. Finally, the time intensity trigeminal effect of (-)-menthol is compared with (1R,2RS,4RS)-1-isopropyl-4-methylbicyclo[3.1.0]hexan-2-ol, named dihydroumbellulol, a new cooling compound obtained by hemi-synthesis from umbellulone extracted from Umbellularia californica Nutt.     

The development of a silica nanoparticle-based label-free DNA biosensor

A silica nanoparticle-based DNA biosensor capable of detecting Bacillus anthracis bacteria through the use of unlabelled ss-oligonucleotides has been developed. The biosensor makes use of the optical changes that accompany a nanoparticle-immobilized cationic conjugated polymer (polythiophene) interacting with single-stranded vs. hybridized oligonucleotides, where a fluorescence signal appears only when hybridized DNA is present (i.e. only when the ss-oligonucleotide interacting with the polymer has hybridized with its complement). In order to enhance the sensitivity of the biosensor, two different nanoparticle architectures were developed and used to elucidate how the presence of neighboring fluorophores on the nanoparticle surface affects F?rster-resonant energy transfer (FRET) between the polythiophene/oligonucleotide complex (FRET donor) and the fluorophores (FRET acceptors). We demonstrate that the silica nanoparticle-based FRET platform lowers the limit of detection at least 10-fold in comparison to the polythiophene itself, and allows the detection of ～2 × 10(-12) moles of ss-oligonucleotide in a 100 μL sample with a standard fluorimeter (i.e. has a limit of detection of ～2 nM ssDNA). Such nanoparticle-based biosensor platforms are beneficial because of the robustness and stability inherent to their covalent assembly and they provide a valuable new tool that may allow for the sensitive, label-free detection (the target DNA that produces the fluorescence signal is unlabelled) without the use of polymerase chain reaction.     

Thermal crosslinking of collagen immobilized on poly(acrylic acid) grafted poly(ethylene terephthalate) films

Graft polymerization of acrylic acid onto plasma‐treated poly(ethylene terephthalate) (PET) films was used to prepare surfaces suitable for collagen immobilization by dip‐coating. Such surfaces could be used as matrices for smooth muscle cell cultures in tissue engineering. Contact angle measurements showed that plasma‐treated and grafted PET films undergo considerable surface reorganization during storage under ambient conditions. However, after collagen immobilization the contact angle remained relatively stable. The amount of collagen initially attached to the film surface increased with increasing poly(acrylic acid) graft density, but subsequent washing in water led to significant collagen loss. This loss could nevertheless be substantially reduced by thermal crosslinking of the collagen in the range 110–130 °C. Atomic force microscopy (AFM) observations suggested that the washed crosslinked collagen has a very similar structure to that of the un‐crosslinked collagen. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 85: 1874–1880, 2002     

PP/EMA TPV: Dynamic cross‐linking through an alcoholysis reaction

Dynamically cross‐linked polypropylene/ethylene‐methyl acrylate copolymer (PP/EMA) blends were prepared by reactive extrusion in a twin‐screw extruder. A transesterification reaction catalyzed by an organotin compound and involving the acrylic moieties of EMA with the hydroxyl functions of pentanediol was used to cross‐link the elastomeric EMA. The extent of cross‐linking was estimated through the insoluble content of the blend. The alcohol‐to‐acrylic ratio and the dibutyltin oxide content were both found to increase the extent of EMA cross‐linking. Processing conditions such as screw rotation rate and extrusion temperature profile also had large effects on the reaction extent. Conditions leading to improved pentanediol dispersion were found to generate larger insoluble fraction. Gel contents of EMA phase as high as 75% to 85% were obtained. Interestingly, the stress‐strain curves of the cross‐linked PP/EMA TPV are very well represented by a single master curve. Dynamically cross‐linked blends show systematically better tensile properties than the non‐reactive system. Blends with higher insoluble content show strain at break in the 650%–750% range. Material processability was maintained, even for the systems with high insoluble content.     

Interaction between dietary protein and fat in triglyceride metabolism in the rat: Effects of soy protein and menhaden oil

The objective of the present study was to determine the mechanisms by which dietary proteins interact with dietary lipids in the regulation of triglyceridemia in rats. Male Sprague-Dawley rats (n=56) were subjected to 28-d experimental diets containing different combinations of proteins (20% w/w) and lipid sources (14% w/w): (i) casein-menhaden oil, (ii) casein-beef tallow, (iii) soy protein-menhaden oil, and (iv) soy protein-beef tallow. Significant protein-lipid interactions were observed on triglyceridemia and hepatic cholesterol in fasted rats. The combination of casein and beef tallow was associated with high plasma TG and hepatic cholesterol concentrations, which were reduced by substitution either of soy for casein or of menhaden oil for beef tallow. Therefore, triglyceridemia and liver cholesterol remained low with soy protein feeding, independently of the lipid source, as well as with menhaden oil feeding, regardless of the protein source. The menhaden oil diets reduced plasma cholesterol, hepatic TG, and TG secretion compared with beef tallow diets independently of the dietary protein source. Modifying the source of dietary proteins and lipids had no effect on post-heparin plasma lipoprotein lipase activity. These results demonstrate that soy protein can lower rat triglyceridemia relative to casein when associated with beef tallow consumption, whereas menhaden oil can attenuate hypertriglyceridemia when rats are fed casein. The data further suggest that part of the hypotriglyceridemic effect of soy protein in the rat may be mediated by reduced hepatic lipid synthesis, as is the case for menhaden oil.     

Hardness, Toughness, and Scratchability of Germanium–Selenium Chalcogenide Glasses

Ge–Se chalcogenide glasses are characterized by relatively low hardness (0.39–2.35 GPa) and low fracture toughness (0.1–0.28 MPa·m^1/2). Actually, the hardness of chalcogen-rich glasses is low enough so that the brittleness parameter, B = H / K _c , is lower than that of silicate glasses. Whereas hardness and Young's modulus increase with increasing germanium contents, fracture toughness follows a trend similar to that of the density and exhibits a maximum for the Ge₂₀Se₈₀ composition, which corresponds to the rigidity percolation threshold. Optical microscopy and atomic force microscopy observations suggest that the indentation deformation proceeds by a localized shear deformation phenomenon. Glasses in the chalcogen-rich region behave viscoelastically at room temperature. As a consequence, an increase of the loading time results in a decrease of hardness and toughness.     

Dietary Intake and Food Sources of Total and Individual Polyunsaturated Fatty Acids in the Belgian Population Over 15 Years Old

Advances in our knowledge of the physiological functions of dietary polyunsaturated fatty acids (PUFAs) have led to an increased interest in food sources and the level of dietary intake of these nutrients. Up to now, no representative data was available for the Belgian adult population. This study aimed to describe data on the intake and food sources of total and individual omega-6 and omega-3 PUFA for the Belgian population over 15 years old. PUFA intakes were assessed for 3,043 Belgian adults, based on two non-consecutive 24 h recalls. Usual intakes were calculated using the multiple source method. The results showed that the intake of linoleic acid (LA) is in accordance with the recommendation for almost all Belgian adults. However, the intake of omega-3 PUFA is suboptimal for a large part of the studied population and also the intake of total PUFA should be increased for a part of the population. The main food source of LA and α-linolenic acid (ALA) was the group of fats and oils (60.6 % for LA and 53.1 % for ALA). Fish and fish products were the most important sources of long chain omega-3 PUFA. Age influenced fatty acids intake, with higher intake of omega-3 PUFA in the older age groups. To fill the gap between the intake and recommendation of total PUFA, and in particular omega-3 PUFA, sustainable strategies and efficient consumer communication strategies will be needed.     

Partial Sperm beta1 Integrin Subunit Deletion Proves Its Involvement in Mouse Gamete Adhesion/Fusion

We have previously shown, using antibodies, that the sperm alpha6beta1 integrin is involved in mouse gamete fusion in vitro. Here we report the conditional knockdown of the sperm Itgb1 gene. It induced a drastic failure of sperm fusogenic ability with sperm accumulation in the perivitelline space of in vitro inseminated oocytes deleted or not for the Itgb1 gene. These data demonstrate that sperm, but not oocyte, beta1 integrin subunit is involved in gamete adhesion/fusion. Curiously, knockdown males were fertile in vivo probably because of the incomplete Cre-mediated deletion of the sperm Itgb1 floxed gene. Indeed, this was shown by Western blot analysis and confirmed by both the viability and litter size of pups obtained by mating partially sperm Itgb1 deleted males with females producing completely deleted Itgb1 oocytes. Because of the total peri-implantation lethality of Itgb1 deletion in mice, we assume that sperm that escaped the Itgb1 excision seemed to be preferentially used to fertilize in vivo. Here, we showed for the first time that the deletion, even partial, of the sperm Itgb1 gene makes the sperm unable to normally fertilize oocytes. However, to elucidate the question of the essentiality of its role during fertilization, further investigations using a mouse expressing a recombinase more effective in male germ cells are necessary.