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.     

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.     

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.     

Dietary intakes and food sources of fatty acids for Belgian women, focused on n-6 and n-3 polyunsaturated fatty acids

The intake of fat, saturated and monounsaturated FA (SFA and MUFA), and omega-6 and omega-3 PUFA has been estimated in 641 Belgian women (age 18-39 y). Their food intake was recorded using a 2-d food diary. The PUFA included were linoleic (LA), alpha-linolenic (LNA), arachidonic (AA), eicosapentaeonoic (EPA), docosapentaenoic (DPA) and docosahexaenoic (DHA) acids. The mean total fat intake corresponded to 34.3% of total energy intake (E). The mean intake of the FA groups corresponded to 13.7%, 13.1%, and 6.0% of E, for SFA, MUFA, and PUFA, respectively. The mean intake of LA was 5.3% of E and of LNA was 0.6% of E, with a mean LA/LNA ratio of 8.7. The mean intake of AA was 0.03% of E. The mean intake of EPA, DPA, and DHA was 0.4%, 0.01%, and 0.06% of E, respectively. According to the Belgian recommendations, the total fat and SFA intake was too high for about three-quarters of the population. The mean LA and overall n-6 PUFA intake corresponded with the recommendation, with part of the population exceeding the upper level. Conversely, the population showed a large deficit for LNA and n-3 PUFA. The major food source for LA and LNA was fats and oils, followed by cereal products. The main sources of long-chain PUFA were fish and seafood, and meat, poultry, and eggs. From a public health perspective, it seems desirable to tackle the problem of low n-3 PUFA intake.     

Fermentation by Amylolytic Lactic Acid Bacteria and Consequences for Starch Digestibility of Plantain, Breadfruit, and Sweet Potato Flours

The potential of tropical starchy plants such as plantain (Musa paradisiaca), breadfruit (Artocarpus communis), and sweet potato (Ipomoea batatas) for the development of new fermented foods was investigated by exploiting the capacity of some lactic acid bacteria to hydrolyze starch. The amylolytic lactic acid bacteria (ALAB) Lactobacillus plantarum A6 and Lactobacillus fermentum Ogi E1 were able to change the consistency of thick sticky gelatinized slurries of these starchy fruits and tubers into semiliquid to liquid products. Consequently, a decrease in apparent viscosity and an increase in Bostwick flow were observed. These changes and the production of maltooligosaccharides confirmed starch hydrolysis. Sucrose in sweet potato was not fermented by strain A6 and poorly fermented by strain Ogi E1, suggesting possible inhibition of sucrose fermentation. In all 3 starchy plants, rapidly digestible starch (RDS) was higher than slowly digestible starch (SDS) and resistant starch (RS) represented between 17% and 30% dry matter (DM). The digestibility of plantain was not affected by fermentation, whereas the RDS content of breadfruit and sweet potato decreased and the RS content increased after fermentation. Practical Application: The characteristics resulting from different combinations of gluten free starchy plants (plantain, breadfruit, sweet potato) and amylolytic lactic acid bacteria (ALAB) offer opportunities to develop new functional fermented beverages, mainly for breadfruit and sweet potato, after further investigation of their formulation, sensory attributes, nutritional, and prebiotic characteristics.     

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.     

Effect of APE1 T2197G (Asp148Glu) Polymorphism on APE1, XRCC1, PARP1 and OGG1 Expression in Patients with Colorectal Cancer

It has been hypothesized that genetic variation in base excision repair (BER) might modify colorectal adenoma risk. Thus, we evaluated the influence of APE1 T2197G (Asp148Glu) polymorphism on APE1, XRCC1, PARP1 and OGG1 expression in normal and tumor samples from patients with colorectal cancer. The results indicate a downregulation of OGG1 and an upregulation of XRCC1 expression in tumor tissue. Regarding the anatomical location of APE1, OGG1 and PARP-1, a decrease in gene expression was observed among patients with cancer in the rectum. In patients with or without some degree of tumor invasion, a significant downregulation in OGG1 was observed in tumor tissue. Interestingly, when taking into account the tumor stage, patients with more advanced grades (III and IV) showed a significant repression for APE1, OGG1 and PARP-1. XRCC1 expression levels were significantly enhanced in tumor samples and were correlated with all clinical and histopathological data. Concerning the polymorphism T2197G, GG genotype carriers exhibited a significantly reduced expression of genes of the BER repair system (APE1, XRCC1 and PARP1). In summary, our data show that patients with colorectal cancer present expression changes in several BER genes, suggesting a role for APE1, XRCC1, PARP1 and OGG1 and APE1 polymorphism in colorectal carcinogenesis.     

Optimal location of piezoelectric actuators for active vibration control of thin axially functionally graded beams

Up to now, optimal location for active control studies concern principally multilayers or homogeneous structures. In the case of functionally graded materials, very few papers exist and they only concern cross section variations. In this way, this paper deals with the optimization of piezoelectric actuators locations on axially functionally graded beams for active vibration control. For this kind of structures, the free vibration problem is more complicated as the governing equations have variable coefficients. Here, the eigenproblem is solved using Fredholm integral equations. The optimal locations of actuators are determined using an optimization criterion, ensuring good controllability of each eigenmode of the structure. The linear quadratic regulator, including a state observer, is used for active control simulations. Two numerical examples are presented for two kinds of boundary conditions.