Product Distributions of Cytochrome P450 OleTJE with Phenyl-Substituted Fatty Acids: A Computational Study

There are two types of cytochrome P450 enzymes in nature, namely, the monooxygenases and the peroxygenases. Both enzyme classes participate in substrate biodegradation or biosynthesis reactions in nature, but the P450 monooxygenases use dioxygen, while the peroxygenases take H₂O₂ in their catalytic cycle instead. By contrast to the P450 monooxygenases, the P450 peroxygenases do not require an external redox partner to deliver electrons during the catalytic cycle, and also no external proton source is needed. Therefore, they are fully self-sufficient, which affords them opportunities in biotechnological applications. One specific P450 peroxygenase, namely, P450 OleT_JE, reacts with long-chain linear fatty acids through oxidative decarboxylation to form hydrocarbons and, as such, has been implicated as a suitable source for the biosynthesis of biofuels. Unfortunately, the reactions were shown to produce a considerable amount of side products originating from C^α and C^β hydroxylation and desaturation. These product distributions were found to be strongly dependent on whether the substrate had substituents on the C^α and/or C^β atoms. To understand the bifurcation pathways of substrate activation by P450 OleT_JE leading to decarboxylation, C^α hydroxylation, C^β hydroxylation and C^α–C^β desaturation, we performed a computational study using 3-phenylpropionate and 2-phenylbutyrate as substrates. We set up large cluster models containing the heme, the substrate and the key features of the substrate binding pocket and calculated (using density functional theory) the pathways leading to the four possible products. This work predicts that the two substrates will react with different reaction rates due to accessibility differences of the substrates to the active oxidant, and, as a consequence, these two substrates will also generate different products. This work explains how the substrate binding pocket of P450 OleT_JE guides a reaction to a chemoselectivity.     

Superlattice and Quantum Dot Unipolar Barrier Infrared Detectors

We report device performance and provide theoretical analysis of a modified long-wavelength complementary barrier infrared detector structure that incorporates a double tunnel junction contact designed for simpler material growth while retaining the robustness for processing. We also provide analysis of a mid-wavelength quantum dot barrier infrared detector and explain its spectral shape and turn-on characteristics.     

Drug Repurposing for Cystic Fibrosis: Identification of Drugs That Induce CFTR-Independent Fluid Secretion in Nasal Organoids

Individuals with cystic fibrosis (CF) suffer from severe respiratory disease due to a genetic defect in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, which impairs airway epithelial ion and fluid secretion. New CFTR modulators that restore mutant CFTR function have been recently approved for a large group of people with CF (pwCF), but ~19% of pwCF cannot benefit from CFTR modulators Restoration of epithelial fluid secretion through non-CFTR pathways might be an effective treatment for all pwCF. Here, we developed a medium-throughput 384-well screening assay using nasal CF airway epithelial organoids, with the aim to repurpose FDA-approved drugs as modulators of non-CFTR-dependent epithelial fluid secretion. From a ~1400 FDA-approved drug library, we identified and validated 12 FDA-approved drugs that induced CFTR-independent fluid secretion. Among the hits were several cAMP-mediating drugs, including β2-adrenergic agonists. The hits displayed no effects on chloride conductance measured in the Ussing chamber, and fluid secretion was not affected by TMEM16A, as demonstrated by knockout (KO) experiments in primary nasal epithelial cells. Altogether, our results demonstrate the use of primary nasal airway cells for medium-scale drug screening, target validation with a highly efficient protocol for generating CRISPR-Cas9 KO cells and identification of compounds which induce fluid secretion in a CFTR- and TMEM16A-indepent manner.     

Polysialic Acid Sustains the Hypoxia-Induced Migration and Undifferentiated State of Human Glioblastoma Cells

Gliomas are the most common primary malignant brain tumors. Glioblastoma, IDH-wildtype (GBM, CNS WHO grade 4) is the most aggressive form of glioma and is characterized by extensive hypoxic areas that strongly correlate with tumor malignancy. Hypoxia promotes several processes, including stemness, migration, invasion, angiogenesis, and radio- and chemoresistance, that have direct impacts on treatment failure. Thus, there is still an increasing need to identify novel targets to limit GBM relapse. Polysialic acid (PSA) is a carbohydrate composed of a linear polymer of α2,8-linked sialic acids, primarily attached to the Neural Cell Adhesion Molecule (NCAM). It is considered an oncodevelopmental antigen that is re-expressed in various tumors. High levels of PSA-NCAM are associated with high-grade and poorly differentiated tumors. Here, we investigated the effect of PSA inhibition in GBM cells under low oxygen concentrations. Our main results highlight the way in which hypoxia stimulates polysialylation in U87-MG cells and in a GBM primary culture. By lowering PSA levels with the sialic acid analog, F-NANA, we also inhibited GBM cell migration and interfered with their differentiation influenced by the hypoxic microenvironment. Our findings suggest that PSA may represent a possible molecular target for the development of alternative pharmacological strategies to manage a devastating tumor like GBM.     

Handedness and "automatization" cognitive style.

Evaluated the relation between "automatization" cognitive style and handedness, or degree of preferred-nonpreferred hand differentiation. 58 male undergraduates were tested on 8 cognitive tasks representing both automatized and nonautomatized abilities. After ipsatization of scores, 14 Ss were chosen as representing "strong automatization," and 14 Ss as "weak automatizers." All Ss were given 6 tests of manual performance using both preferred and nonpreferred hands. No differences were found between strong and weak automatizers in the degree of differentiation between overall preferred and nonpreferred hand performance. Explanations for these findings and the possible role of automatization cognitive style in hand differentiation in young children are discussed. (French summary) (15 ref.) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

MLS‐based variable‐node elements compatible with quadratic interpolation. Part II: application for finite crack element

Two‐dimensional finite ‘crack’ elements for simulation of propagating cracks are developed using the moving least‐square (MLS) approximation. The mapping from the parental domain to the physical element domain is implicitly obtained from MLS approximation, with the shape functions and their derivatives calculated and saved only at the numerical integration points. The MLS‐based variable‐node elements are extended to construct the crack elements, which allow the discontinuity of crack faces and the crack‐tip singularity. The accuracy of the crack elements is checked by calculating the stress intensity factor under mode I loading. The crack elements turn out to be very efficient and accurate for simulating crack propagations, only with the minimal amount of element adjustment and node addition as the crack tip moves. Numerical results and comparison to the results from other works demonstrate the effectiveness and accuracy of the present scheme for the crack elements. Copyright © 2005 John Wiley & Sons, Ltd.     

Morphological characterization of sulfonated graphene and Nafion composite membrane by dynamic mode atomic force microscopy

Maintaining proper hydration in the proton exchange membrane (PEM) is a crucial issue for passive air breathing PEM (ABPEM) fuel cells. The inorganic filler Nafion composite membrane has great potential for replacing commercialized PEM for ABPEM. We synthesize sulfonated graphene and Nafion composite membrane by using functionalized graphene to enhance the water content in the membrane. We analyze morphological variation of a sulfonated graphene–Nafion composite membrane by using dynamic mode atomic force microscopy (DMAFM). The phase map and topography of the sulfonated graphene–Nafion composite membrane were simultaneously studied for systematic characterization. Through characterization, we find that the water content on the composite membrane has increased remarkably and that it is related to the morphological variation after the composition. In the composite membrane, the root‐mean‐squared surface roughness has increased compared with pristine Nafion because of the sulfonated graphene. In the DMAFM study, the composite membrane shows an entirely different phase map than the pristine Nafion. The elliptical domains, which have positive phase lags, are created on the composite membrane surface. The creation of these domains reflects the existence of a repulsive interaction between the tip and sample surface because of an increasing adhesive force between the tip and sample. Copyright © 2015 John Wiley & Sons, Ltd.     

Effect of Heat Input on Macro,Micro and Tensile Properties of Flux Cored Arc Welded Ferritic Stainless Steel Joints

This paper discusses the influence of Flux Cored Arc Welding Process parameters such as welding current, travel speed, voltage on bead profile, metallurgical and mechanical properties of welds of 2 mm thick 409M ferritic stainless steel sheets. The study reveals that, grain coarsening, volume fraction of martensite, hardness of heat affected zone and % of delta ferrite in ER 309 weld metal increases with increase in heat input. However, the results show that variation of heat input does not make any significant effect on tensile strength of the joint. Hence, welding parameters that provide uniform bead profile for the weld are recommended for fabrication.     

A tractable genotype–phenotype map modelling the self-assembly of protein quaternary structure

The mapping between biological genotypes and phenotypes is central to the study of biological evolution. Here, we introduce a rich, intuitive and biologically realistic genotype–phenotype (GP) map that serves as a model of self-assembling biological structures, such as protein complexes, and remains computationally and analytically tractable. Our GP map arises naturally from the self-assembly of polyomino structures on a two-dimensional lattice and exhibits a number of properties: redundancy (genotypes vastly outnumber phenotypes), phenotype bias (genotypic redundancy varies greatly between phenotypes), genotype component disconnectivity (phenotypes consist of disconnected mutational networks) and shape space covering (most phenotypes can be reached in a small number of mutations). We also show that the mutational robustness of phenotypes scales very roughly logarithmically with phenotype redundancy and is positively correlated with phenotypic evolvability. Although our GP map describes the assembly of disconnected objects, it shares many properties with other popular GP maps for connected units, such as models for RNA secondary structure or the hydrophobic-polar (HP) lattice model for protein tertiary structure. The remarkable fact that these important properties similarly emerge from such different models suggests the possibility that universal features underlie a much wider class of biologically realistic GP maps.