Towards accurate numerical method for monodomain models using a realistic heart geometry

The simulation of cardiac electrophysiological waves are known to require extremely fine meshes, limiting the applicability of current numerical models to simplified geometries and ionic models. In this work, an accurate numerical method based on a time-dependent anisotropic remeshing strategy is presented for simulating three-dimensional cardiac electrophysiological waves. The proposed numerical method greatly reduces the number of elements and enhances the accuracy of the prediction of the electrical wave fronts. Illustrations of the performance and the accuracy of the proposed method are presented using a realistic heart geometry. Qualitative and quantitative results show that the proposed methodology is far superior to the uniform mesh methods commonly used in cardiac electrophysiology.     

Duplication of Horizontally Acquired GH5_2 Enzymes Played a Central Role in the Evolution of Longhorned Beetles

The rise of functional diversity through gene duplication contributed to the adaption of organisms to various environments. Here we investigate the evolution of putative cellulases of the subfamily 2 of glycoside hydrolase family 5 (GH5_2) in the Cerambycidae (longhorned beetles), a megadiverse assemblage of mostly xylophagous beetles. Cerambycidae originally acquired GH5_2 from a bacterial donor through horizontal gene transfer (HGT), and extant species harbor multiple copies that arose from gene duplication. We ask how these digestive enzymes contributed to the ability of these beetles to feed on wood. We analyzed 113 GH5_2, including the functional characterization of 52 of them, derived from 25 species covering most subfamilies of Cerambycidae. Ancestral gene duplications led to five well-defined groups with distinct substrate specificity, allowing these beetles to break down, in addition to cellulose, polysaccharides that are abundant in plant cell walls (PCWs), namely, xyloglucan, xylan, and mannans. Resurrecting the ancestral enzyme originally acquired by HGT, we show it was a cellulase that was able to break down glucomannan and xylan. Finally, recent gene duplications further expanded the catalytic repertoire of cerambycid GH5_2, giving rise to enzymes that favor transglycosylation over hydrolysis. We suggest that HGT and gene duplication, which shaped the evolution of GH5_2, played a central role in the ability of cerambycid beetles to use a PCW-rich diet and may have contributed to their successful radiation.     

Discriminating mild from critical COVID-19 by innate and adaptive immune single-cell profiling of bronchoalveolar lavages

How the innate and adaptive host immune system miscommunicate to worsen COVID-19 immunopathology has not been fully elucidated. Here, we perform single-cell deep-immune profiling of bronchoalveolar lavage (BAL) samples from 5 patients with mild and 26 with critical COVID-19 in comparison to BALs from non-COVID-19 pneumonia and normal lung. We use pseudotime inference to build T-cell and monocyte-to-macrophage trajectories and model gene expression changes along them. In mild COVID-19, CD8⁺ resident-memory (T_RM) and CD4⁺ T-helper-17 (T_H17) cells undergo active (presumably antigen-driven) expansion towards the end of the trajectory, and are characterized by good effector functions, while in critical COVID-19 they remain more naïve. Vice versa, CD4⁺ T-cells with T-helper-1 characteristics (T_H1-like) and CD8⁺ T-cells expressing exhaustion markers (T_EX-like) are enriched halfway their trajectories in mild COVID-19, where they also exhibit good effector functions, while in critical COVID-19 they show evidence of inflammation-associated stress at the end of their trajectories. Monocyte-to-macrophage trajectories show that chronic hyperinflammatory monocytes are enriched in critical COVID-19, while alveolar macrophages, otherwise characterized by anti-inflammatory and antigen-presenting characteristics, are depleted. In critical COVID-19, monocytes contribute to an ATP-purinergic signaling-inflammasome footprint that could enable COVID-19 associated fibrosis and worsen disease-severity. Finally, viral RNA-tracking reveals infected lung epithelial cells, and a significant proportion of neutrophils and macrophages that are involved in viral clearance.Subject terms: Genome-wide analysis of gene expression, Innate immunity, Bioinformatics     

Structural,mutagenic and in silico studies of xyloglucan fucosylation in Arabidopsis thaliana suggest a water‐mediated mechanism

The mechanistic underpinnings of the complex process of plant polysaccharide biosynthesis are poorly understood, largely because of the resistance of glycosyltransferase (GT) enzymes to structural characterization. In Arabidopsis thaliana, a glycosyl transferase family 37 (GT37) fucosyltransferase 1 (AtFUT1) catalyzes the regiospecific transfer of terminal 1,2‐fucosyl residues to xyloglucan side chains – a key step in the biosynthesis of fucosylated sidechains of galactoxyloglucan. We unravel the mechanistic basis for fucosylation by AtFUT1 with a multipronged approach involving protein expression, X‐ray crystallography, mutagenesis experiments and molecular simulations. Mammalian cell culture expressions enable the sufficient production of the enzyme for X‐ray crystallography, which reveals the structural architecture of AtFUT1 in complex with bound donor and acceptor substrate analogs. The lack of an appropriately positioned active site residue as a catalytic base leads us to propose an atypical water‐mediated fucosylation mechanism facilitated by an H‐bonded network, which is corroborated by mutagenesis experiments as well as detailed atomistic simulations.     

Heterologous expression of a β-<Emphasis Type="SmallCaps">d</Emphasis>-glucosidase in <Emphasis Type="Italic">Caldicellulosiruptor bescii</Emphasis> has a surprisingly modest effect on the activity of the exoproteome and growth on crystalline cellulose

Members of the genus Caldicellulosiruptor are the most thermophilic cellulolytic bacteria so far described and are capable of efficiently utilizing complex lignocellulosic biomass without conventional pretreatment. Previous studies have shown that accumulation of high concentrations of cellobiose and, to a lesser extent, cellotriose, inhibits cellulase activity both in vivo and in vitro and high concentrations of cellobiose are present in C. bescii fermentations after 90 h of incubation. For some cellulolytic microorganisms, β-d-glucosidase is essential for the efficient utilization of cellobiose as a carbon source and is an essential enzyme in commercial preparations for efficient deconstruction of plant biomass. In spite of its ability to grow efficiently on crystalline cellulose, no extracellular β-d-glucosidase or its GH1 catalytic domain could be identified in the C. bescii genome. To investigate whether the addition of a secreted β-d-glucosidase would improve growth and cellulose utilization by C. bescii, we cloned and expressed a thermostable β-d-glucosidase from Acidothermus cellulolyticus (Acel_0133) in C. bescii using the CelA signal sequence for protein export. The effect of this addition was modest, suggesting that β-d-glucosidase is not rate limiting for cellulose deconstruction and utilization by C. bescii.     

Insulin effect upon lipogenesis of perfused mouse liver

Lipogenesis of mouse livers was estimated by the incorporation of tritium, from triated water, into triglyceride fatty acids, thus eliminating the problem of varying specific activity of metabolic precursors which is met when using ¹⁴C- or ³H-labelled substrates. Using this procedure, a rapid (within 2 h) stimulatory effect of insulin upon lipogenesis of perfused livers obtained from anti-insulin serum-treated normal or obese-hyperglycemic (ob/ob) micr has been observed. Anti-insulin serum treatment did not alter hepatic glycogen or cyclic AMP content. The smallest effective stimulatory concentration of the hormone was 50 micro unit per ml. Insulin increased lipogenesis in the presence of either glucose or acetate but not in the absence of substrate. It did not relieve the inhibitory effect of added oleate upon the lipogenic process. The clear-cut stimulator effect of insulin upon lipogenesis observed in livers from anti-insulin serum-treated normal or obese-hyperglycemic mice was no longer present when livers from untreated animals were studied. Under the latter conditions, basal lipogenesis was higher than that seen in livers of animals treated with anti-insulin serum prior to the experiment, being highest in livers obtained from hyperinsulinemic obese-hyperglycemic mice. This suggested that the presence of endogenous insulin immediately prior to the experimental sufficed to stimulate hepatic lipogenesis, the degree of this stimulation being apparently related to that of insulinemia. Although the precise site of aciton of the rapid stimulatory influence of insulin upon liver lipogenesis is not determined, it appears to be situated at or beyond the level of acetyl-CoA carboxylase, fatty acid handling and cyclic AMP being apparently not implicated in such hormonal regulations.     

Random amplified polymorphic DNA and pedigree relationships in spring barley

Summary We investigated random amplified polymorphic DNA (RAPD) in 27 inbred barley lines with varying amounts of common ancestry and in 20 doubled-haploid (DH) lines from a biparental cross. Of 33 arbitrary 10 base primers that were tested, 19 distinguished a total of 31 polymorphisms. All polymorphisms were scored as dominant genetic markers except for 1, where Southern analysis indicated the presence of two codominant amplification products. The inheritance of 19 RAPD polymorphisms and one morphological trait was studied in the DH lines. There was no evidence for segregation distortion, but a group of four tightly linked loci was detected. The frequencies of RAPD polymorphism in pairs of inbred lines were used to compute values of genetic distance (d), which were compared to kinship coefficients (r) between the same pairs of lines. A linear relationship between r and d was evident, but low values of r gave poor predictions of d. Cluster analysis showed that groups of inbred lines based on r were similar to those based on d with some notable exceptions. RAPD markers can be used to gain information about genetic similarities or differences that are not evident from pedigree information.