Effect of Oral and Parenteral N Nutrition vs N-Free Nutrition on the Endogenous Amino Acid Flow at the Ileum of the Pig*

Two methods were tested for suppressing the depressive effect of N-free diets on the digestive secretions in pigs: the blood perfusion of amino acids (AA) or the peptide alimentation method. In the latter, enzymically hydrolysed casein (EHC), composed of oligopeptides and free AA, was used as the source of nitrogen. The unabsorbed dietary N molecules were discarded from the ileal digesta by ultrafiltration or gel filtration, assuming that the endogenous fraction did not contain significant amounts of small molecules. The AA supply by blood perfusion had no effect on the ileal endogenous AA losses (8·0 g AA kg⁻¹ DM intake) in growing pigs (±50 kg), compared with the N-free diet alone (8·3 g), whereas the EHC supplementation significantly increased them (18·0 g). The increase was due to both endogenous and dietary N. The presence of unabsorbed dietary AA in the ileal digesta was confirmed by the AA profile of the soluble molecules with a very low molecular mass (<3 kDa), which was close to that of EHC. Both ultrafiltration (cut-offs of 3 or 10 kDa) and gel filtration methods, utilised to discard the remaining dietary molecules, also eliminated a significant proportion of endogenous AA.     

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.     

Fusarium graminearum Infection Strategy in Wheat Involves a Highly Conserved Genetic Program That Controls the Expression of a Core Effectome

Fusarium graminearum, the main causal agent of Fusarium Head Blight (FHB), is one of the most damaging pathogens in wheat. Because of the complex organization of wheat resistance to FHB, this pathosystem represents a relevant model to elucidate the molecular mechanisms underlying plant susceptibility and to identify their main drivers, the pathogen’s effectors. Although the F. graminearum catalog of effectors has been well characterized at the genome scale, in planta studies are needed to confirm their effective accumulation in host tissues and to identify their role during the infection process. Taking advantage of the genetic variability from both species, a RNAseq-based profiling of gene expression was performed during an infection time course using an aggressive F. graminearum strain facing five wheat cultivars of contrasting susceptibility as well as using three strains of contrasting aggressiveness infecting a single susceptible host. Genes coding for secreted proteins and exhibiting significant expression changes along infection progress were selected to identify the effector gene candidates. During its interaction with the five wheat cultivars, 476 effector genes were expressed by the aggressive strain, among which 91% were found in all the infected hosts. Considering three different strains infecting a single susceptible host, 761 effector genes were identified, among which 90% were systematically expressed in the three strains. We revealed a robust F. graminearum core effectome of 357 genes expressed in all the hosts and by all the strains that exhibited conserved expression patterns over time. Several wheat compartments were predicted to be targeted by these putative effectors including apoplast, nucleus, chloroplast and mitochondria. Taken together, our results shed light on a highly conserved parasite strategy. They led to the identification of reliable key fungal genes putatively involved in wheat susceptibility to F. graminearum, and provided valuable information about their putative targets.     

Rheology evolution of a geopolymer precursor aqueous suspension during aging

Geopolymer-based glass-ceramic matrix composites can be processed at room temperature and a heat treatment below 100°C leads to matrix hardening thanks to the geopolymerization mechanisms. The stabilization of the matrix into glass-ceramics is achieved via a post-curing at high temperature. This paves the way of the utilization of cost-effective liquid composite molding processes, for which all the necessary equipment is already available for processing temperature ranges related to polymer matrix composites, provided that the rheological behavior of the precursor is suitable to conveniently permeate the fibrous preform. The paper describes the thixotropic rheological behavior of a reference suspension at processing temperature (10°C-20°C) and its evolution along aging at −18°C. The changes are interpreted in terms of geopolymerization mechanisms (dissolution and polycondensation) and suspension rheology (predominance of hydrodynamic effects at high shear rate). On this basis, a phenomenological modeling framework, combining two Krieger-Dougherty equations, is proposed to build a relationship between the effective viscosity of the suspension and the phenomena involved during aging (dissolution of aluminosilicate particles) and shearing (microstructure scalar variable).     

Application of ATR-FTIR for a rapid and simultaneous determination of sugars and organic acids in apricot fruit

A simple, fast and accurate Fourier transform mid-infrared spectroscopy method was developed for simultaneously determining sugar and organic acid contents in apricot fruit slurries using the attenuated total reflectance. The potential of this method coupled with chemometric techniques based on partial least squares was assessed by comparison with currently used enzymatic determination of sucrose, glucose, fructose, malic acid and citric acid. Fruits of eight contrasted cultivars harvested at different ripening stages were used in this study and randomly divided in a calibration set (505 apricots) and in a validation set (252 apricots). The most suitable region was found in the range between 1500 and 900 cm⁻¹. Good prediction performances were obtained (R² ? 0.74 and RMSEP ? 18%). Results concerning the prediction of other quality traits such as firmness, skin colour, ethylene production, soluble solids content and titratable acidity were discussed.     

Rapid and non-destructive analysis of apricot fruit quality using FT-near-infrared spectroscopy

A non-destructive optical method based on near-infrared spectroscopy has been used for the evaluation of apricot fruit quality. Diffuse reflectance measurements (800–2500 nm), physical, physiological and biochemical measurements were performed individually on 877 apricot fruits from eight contrasted cultivars harvested at different ripening stages. Relationships between spectral wavelengths and quality attributes were evaluated by application of chemometric techniques based on partial least squares (PLS) on fruit set divided randomly into two groups: 598 fruits for calibration and 279 for validation. Good prediction performance was obtained for soluble solids and titratable acidity with correlation coefficients of 0.92 and 0.89 respectively and root mean square errors of prediction of 0.98% Brix and 3.62 meq 100 g⁻¹ FW respectively. For the other quality traits such as firmness, ethylene, individual sugars and organic acids, the prediction models were not satisfactorily accurate due to the high error of calibration and prediction.     

A two-step cloning-free PCR-based method for the deletion of genes in the opportunistic pathogenic yeast Candida lusitaniae

We describe a new cloning-free strategy to delete genes in the opportunistic pathogenic yeast Candida lusitaniae. We first constructed two ura3 Δ strains in C. lusitaniae for their use in transformation experiments. One was deleted for the entire URA3 coding sequence; the other possessed a partial deletion within the coding region, which was used to determine the minimum amount of homology required for efficient homologous recombination by double crossing-over of a linear DNA fragment restoring URA3 expression. This amount was estimated to 200 bp on each side of the DNA fragment. These data constituted the basis of the development of a strategy to construct DNA cassettes for gene deletion by a cloning-free overlapping PCR method. Two cassettes were necessary in two successive transformation steps for the complete removal of a gene of interest. As an example, we report here the deletion of the LEU2 gene. The first cassette was constituted by the URA3 gene flanked by two large fragments (500 bp) homologous to the 5' and 3' non-coding regions of LEU2. After transformation of an ura3 Δ recipient strain and integration of the cassette at the LEU2 locus, the URA3 gene was removed by a second transformation round with a DNA cassette made by the fusion between the 5' and 3' non-coding regions of the LEU2 gene. The overall procedure takes less than 2 weeks and allows the creation of a clean null mutant that retains no foreign DNA sequence integrated in its genome.     

Effects of the smoking process on odour characteristics of smoked herring (Clupea harengus) and relationships with phenolic compound content

The relationship between smoking parameters and odour characteristics, evaluated by a trained sensory panel, were studied on smoked herring. In addition, a possible correlation between the content of 10 phenolic compounds and sensory perceptions was investigated. Five smoking techniques were applied, combining smoke production conditions, performed by pyrolysis of beech wood sawdust or by friction of beech wood log, with smoke deposition, either in a controlled kiln (traditional smoking) or by an electrostatic process. In the fifth smoking technique, a purified condensate of beech smokes was vaporised on fish fillets in the smokehouse. The time of smoking was 3 h for traditional smoking and the liquid smoke atomisation process and 12 min for the electrostatic method. The effects of three smoking temperatures (16, 24, 32 °C) were tested for both the traditional and the liquid smoke atomisation processes, as well as the effect of the position of the exhaust valve in the smokehouse in the case of the traditional method. Two different voltages were applied for the electrostatic process, 37 and 42 kV.