Heads or Tails: Genotyping of Hepatitis C Virus Concerning the 2k/1b Circulating Recombinant Form

As different hepatitis C virus (HCV) genotypes respond differently to initiated therapy, correct HCV genotyping is essential. A potential risk for misclassification of the intergenotypic HCV circulating recombinant form (CRF) 2k/1b strains exists, depending on the genotyping method used. The aim was to investigate the differences in HCV genotyping methods with regard to CRF 2k/1b and to gain insight in the prevalence of the CRF 2k/1b. Genotyping results by Versant HCV Genotype Assay were compared with nonstructural protein 5B (NS5B) sequencing. In total, from November 2001 until March 2015, 3296 serum samples were analyzed by Versant HCV Genotype Assay. As misclassified CRF is harbored among HCV genotype 2, we further focused our search on 142 (4.3%) samples positive for HCV genotype 2. On 116 (81.7%) retrieved samples, the NS5B sequencing was performed. Twelve out of the 116 retrieved samples (10.3%) were classified as CRF 2k/1b by sequencing of the NS5B region. Ten of these 12 samples were originally misclassified as genotype 2a or 2c, while 2 of them were misclassified as genotype 2. Our results show that the current prevalence of CRF 2k/1b is underestimated. The importance of correct HCV genotyping is emphasized, considering the tailored choice of treatment regimen and overall prognosis.     

Mechanical and barrier properties of cardboard and 3D packaging coated with microfibrillated cellulose

Microfibrillated cellulose (MFC) is increasingly used with cellulosic substrates and especially with paper materials. Its use with cardboard remains not reported and the study of mechanical and barrier properties of MFC‐coated cardboard has been investigated in this article. The influence of coating process as well as the effect of MFC have been highlighted by comparing different MFC‐coated cardboard samples with PE‐coated cardboard samples. MFC was coated using bar coating process. Their distribution and homogeneity onto cardboard was observed using techniques such as SEM and FE‐SEM. Tests such as oxygen and air permeability, bending stiffness, and compressive strength have been carried out. The coating process used impacts significantly cardboard properties by two opposite ways: on one hand it damages the structure cohesion of cardboard decreasing its compressive strength; on the other hand it increases its bending stiffness by increasing considerably the samples thickness. The addition of MFC counterbalances the negative effects of the coating process: bending stiffness and compressive strength are indeed improved by 30% in machine direction. On the contrary, MFC does not enhance much cardboard barrier properties, although it considerably increases their water absorption. Within a framework of packaging application, MFC will rather have consequent effects on cardboard's properties as blend or as part of the multilayer structure. Other applications have to be considered for its use as top layer. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40106.     

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.     

A New Strategy to Preserve and Assess Oxygen Consumption in Murine Tissues

Mitochondrial dysfunctions are implicated in several pathologies, such as metabolic, cardiovascular, respiratory, and neurological diseases, as well as in cancer and aging. These metabolic alterations are usually assessed in human or murine samples by mitochondrial respiratory chain enzymatic assays, by measuring the oxygen consumption of intact mitochondria isolated from tissues, or from cells obtained after physical or enzymatic disruption of the tissues. However, these methodologies do not maintain tissue multicellular organization and cell-cell interactions, known to influence mitochondrial metabolism. Here, we develop an optimal model to measure mitochondrial oxygen consumption in heart and lung tissue samples using the XF24 Extracellular Flux Analyzer (Seahorse) and discuss the advantages and limitations of this technological approach. Our results demonstrate that tissue organization, as well as mitochondrial ultrastructure and respiratory function, are preserved in heart and lung tissues freshly processed or after overnight conservation at 4 °C. Using this method, we confirmed the repeatedly reported obesity-associated mitochondrial dysfunction in the heart and extended it to the lungs. We set up and validated a new strategy to optimally assess mitochondrial function in murine tissues. As such, this method is of great potential interest for monitoring mitochondrial function in cohort samples.     

A Deep Dive into VDAC1 Conformational Diversity Using All-Atom Simulations Provides New Insights into the Structural Origin of the Closed States

The voltage-dependent anion channel 1 (VDAC1) is a crucial mitochondrial transporter that controls the flow of ions and respiratory metabolites entering or exiting mitochondria. As a voltage-gated channel, VDAC1 can switch between a high-conducting “open” state and a low-conducting “closed” state emerging at high transmembrane (TM) potentials. Although cell homeostasis depends on channel gating to regulate the transport of ions and metabolites, structural hallmarks characterizing the closed states remain unknown. Here, we performed microsecond accelerated molecular dynamics to highlight a vast region of VDAC1 conformational landscape accessible at typical voltages known to promote closure. Conformers exhibiting durable subconducting properties inherent to closed states were identified. In all cases, the low conductance was due to the particular positioning of an unfolded part of the N-terminus, which obstructed the channel pore. While the N-terminal tail was found to be sensitive to voltage orientation, our models suggest that stable low-conducting states of VDAC1 predominantly take place from disordered events and do not result from the displacement of a voltage sensor or a significant change in the pore. In addition, our results were consistent with conductance jumps observed experimentally and corroborated a recent study describing entropy as a key factor for VDAC gating.     

SMART-Reactors: Tailoring Gas Holdup Distribution by Additively Manufactured Lattice Structures

In chemical process engineering, fast gas-liquid reactions often suffer from an inefficient distribution of gas and therefore mixing and mass transfer performance. This study deals with the possibility of influencing the local gas holdup and bubble size distribution in a gas-liquid process using additively manufactured lattice structures (AMLS). The used measuring technique to study bubble size, velocity, and the local gas holdup is a photo-optical needle probe. By using AMLS, a significant radial homogenization of the local gas holdup and the mean bubble size is achieved. Furthermore, it can be demonstrated that the bubble size can be tailored by the geometry of the inserted structure. It is illustrated that the mean bubble velocities are lowered within the lattice resulting in a higher residence time of the dispersed phase with an impact on the mass transfer performance within the AMLS.     

Label-Free Study of the Global Cell Behavior during Exposure to Environmental Radiofrequency Fields in the Presence or Absence of Pro-Apoptotic or Pro-Autophagic Treatments

It remains controversial whether exposure to environmental radiofrequency signals (RF) impacts cell status or response to cellular stress such as apoptosis or autophagy. We used two label-free techniques, cellular impedancemetry and Digital Holographic Microscopy (DHM), to assess the overall cellular response during RF exposure alone, or during co-exposure to RF and chemical treatments known to induce either apoptosis or autophagy. Two human cell lines (SH-SY5Y and HCT116) and two cultures of primary rat cortex cells (astrocytes and co-culture of neurons and glial cells) were exposed to RF using an 1800 MHz carrier wave modulated with various environmental signals (GSM: Global System for Mobile Communications, 2G signal), UMTS (Universal Mobile Telecommunications System, 3G signal), LTE (Long-Term Evolution, 4G signal, and Wi-Fi) or unmodulated RF (continuous wave, CW). The specific absorption rates (S.A.R.) used were 1.5 and 6 W/kg during DHM experiments and ranged from 5 to 24 W/kg during the recording of cellular impedance. Cells were continuously exposed for three to five consecutive days while the temporal phenotypic signature of cells behavior was recorded at constant temperature. Statistical analysis of the results does not indicate that RF-EMF exposure impacted the global behavior of healthy, apoptotic, or autophagic cells, even at S.A.R. levels higher than the guidelines, provided that the temperature was kept constant.     

The effects of soaking of whole, dehulled and ground millet and soybean seeds on phytate degradation and Phy/Fe and Phy/Zn molar ratios

The degradation and leaching of phytates, phytase activity and iron and zinc concentrations have been studied after soaking of whole seeds, dehulled seeds and flours of millet and soybean, in order to investigate the efficiency of soaking on reducing Phy/Fe and Phy/Zn molar ratios. When using millet grains, dehulling and milling before soaking facilitated the leaching of phytates and phytases in the aqueous medium and hence phytate degradation. Dehulling of soybean seeds led to a marked increase in phytate content, whereas milling favoured reactions between phytases and phytates. The Phy/Fe and Phy/Zn molar ratios decreased only slightly during soaking. The highest decreases for millet were obtained after soaking of flour for 8 h (Phy/Fe: 10.8–7.7 and Phy/Zn: 20.3–15.1), and after soaking of whole seeds for 24 h for soybean (Phy/Fe: 10.4–9.4 and Phy/Zn: 23.8–19.1). Cooking of flours with water used for soaking did not increase phytate degradation.     

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.