Characterization of a human coagulation factor Xa-binding site on Viperidae snake venom phospholipases A2 by affinity binding studies and molecular bioinformatics

<p>Abstract</p> <p>Background</p> <p>The snake venom group IIA secreted phospholipases A₂(SVPLA₂), present in the <it>Viperidae </it>snake family exhibit a wide range of toxic and pharmacological effects. They exert their different functions by catalyzing the hydrolysis of phospholipids (PL) at the membrane/water interface and by highly specific direct binding to: (i) presynaptic membrane-bound or intracellular receptors; (ii) natural PLA₂-inhibitors from snake serum; and (iii) coagulation factors present in human blood.</p> <p>Results</p> <p>Using surface plasmon resonance (SPR) protein-protein interaction measurements and an <it>in vitro </it>biological test of inhibition of prothrombinase activity, we identify a number of <it>Viperidae </it>venom SVPLA₂s that inhibit blood coagulation through direct binding to human blood coagulation factor Xa (FXa) via a non-catalytic, PL-independent mechanism. We classify the SVPLA₂s in four groups, depending on the strength of their binding.</p> <p>Molecular electrostatic potentials calculated at the surface of 3D homology-modeling models show a correlation with inhibition of prothrombinase activity. In addition, molecular docking simulations between SVPLA₂and FXa guided by the experimental data identify the potential FXa binding site on the SVPLA₂s. This site is composed of the following regions: helices A and B, the Ca²⁺loop, the helix C-β-wing loop, and the C-terminal fragment. Some of the SVPLA₂binding site residues belong also to the interfacial binding site (IBS). The interface in FXa involves both, the light and heavy chains.</p> <p>Conclusion</p> <p>We have experimentally identified several strong FXa-binding SVPLA₂s that disrupt the function of the coagulation cascade by interacting with FXa by the non-catalytic PL-independent mechanism. By theoretical methods we mapped the interaction sites on both, the SVPLA₂s and FXa. Our findings may lead to the design of novel, non-competitive FXa inhibitors.</p

Spin dependent photoelectron tunnelling from GaAs into magnetic Cobalt

The spin dependence of the photoelectron tunnel current from free standing GaAs films into out-of- plane magnetized Cobalt films is demonstrated. The measured spin asymmetry (A) resulting from a change in light helicity, reaches +/- 6% around zero applied tunnel bias and drops to +/- 2% at a bias of -1.6 V applied to the GaAs. This decrease is a result of the drop in the photoelectron spin polarization that results from a reduction in the GaAs surface recombination velocity. The sign of A changes with that of the Cobalt magnetization direction. In contrast, on a (nonmagnetic) Gold film A ~ 0%

Enhanced Phenotype Definition for Precision Isolation of Precursor Exhausted Tumor-Infiltrating CD8 T Cells.

In the context of adoptive T cell transfer (ACT) for cancer treatment, it is crucial to generate in vitro large amounts of tumor-specific CD8 T cells with high potential to persist in vivo. PD-1, Tim3, and CD39 have been proposed as markers of tumor-specific tumor-infiltrating CD8 T lymphocytes (CD8 TILs). However, these molecules are highly expressed by terminally differentiated exhausted CD8 T cells (Tex) that lack proliferation potential. Therefore, optimized strategies to isolate tumor-specific TILs with high proliferative potential, such as Tcf1+ precursor exhausted T cells (Tpe) are needed to improve in vivo persistence of ACT. Here we aimed at defining cell surface markers that would unequivocally identify Types for precision cell sorting increasing the purity of tumor-specific PD-1+ Tcf1+ Tpe from total TILs. Transcriptomic analysis of Tpe vs. Tex CD8 TIL subsets from B16 tumors and primary human melanoma tumors revealed that Tpes are enriched in Slamf6 and lack Entpd1 and Havcr2 expression, which encode Slamf6, CD39, and Tim3 cell surface proteins, respectively. Indeed, we observed by flow cytometry that CD39- Tim3- Slamf6+ PD-1+ cells yielded maximum enrichment for tumor specific PD-1+ Tcf1+ OT1 TILs in B16.OVA tumors. Moreover, this population showed higher re-expansion capacity upon an acute infection recall response compared to the CD39+ counterparts or bulk PD-1+ TILs. Hence, we report an enhanced sorting strategy (CD39- Tim3- Slamf6+ PD-1+) of Tpes. In conclusion, we show that optimization of CD8 TIL cell sorting strategy is a viable approach to improve recall capacity and in vivo persistence of transferred cells in the context of ACT

Framework for academic success: Roles and responsibilities of both students and lecturers

This book aims to advance the knowledge on the future of business education in South Africa and to allow all relevant role players (universities and industry) the opportunity to debate and share ideas on how to best position business education to optimally serve the interest of students and the dynamic changes taking place in the world of work. Although some of these changes have taken shape before 2020, the COVID-19 pandemic has accelerated the need and pace for change. If business schools (in our context, faculties of economic and management sciences) do not adapt rapidly, they will be left behind by other up-and-coming industry providers. The research scope covers all business-related undergraduate and postgraduate economics, management and accounting programmes, excluding MBA programmes

Strategy disclosures by listed financial services companies: Signalling theory, legitimacy theory and South African integrated reporting practices

This paper investigates the current extent of social-, environmental- and economic- (SEE) related strategy disclosure of companies listed in the financial services sector of the Johannesburg Stock Exchange. It uses signalling and legitimacy theory to analyse the findings from a developed SEE strategy-related disclosure checklist. The paper finds social and environmental strategy-related disclosure is still secondary to economic strategy-related disclosures. This may be due to persistent focus on providers of financial capital and the need to perform financially. Further, the subsector’s business model and how closely the subsector interacts with their customers is seen as a driver of social and environmental strategy-related disclosure to maintain their legitimacy and to reduce information asymmetry, reduce cost of capital and assure investors that these factors are being appropriately managed by the entity. Following from above, the banking, insurance and real estate subsectors presented the most strategy-related disclosure. This was linked to their high public accountability and daily interaction with customers, necessitating the need to manage their legitimacy and address adverse selection. The paper also proposes some areas for future research to understand the potential obstacles to incorporating social and environmental concerns into strategy and related disclosures

The effect of completeness of revascularization during CABG with single versus multiple arterial grafts

IntroductionIncomplete coronary revascularization is associated with suboptimal outcomes. We investigated the longâ term effects of Incomplete, Complete, and Supraâ complete revascularization and whether these effects differed in the setting of singleâ arterial and multiâ arterial coronary artery bypass graft (CABG).MethodsWe analyzed 15â year mortality in 7157 CABG patients (64.1â ±â 10.5 years; 30% women). All patients received a left internal thoracic artery to left anterior descending coronary artery graft with additional venous grafts only (singleâ arterial) or with at least one additional arterial graft (multiâ arterial) and were grouped based on a completeness of revascularization index (CRIâ =â number of grafts minus the number of diseased principal coronary arteries): Incomplete (CRIâ â ¤â â 1 [Nâ =â 320;4.5%]); Complete (CRIâ =â 0 [Nâ =â 2882;40.3%]; reference group); and two Supraâ complete categories (CRIâ =â +1[Nâ =â 3050; 42.6%]; CRIâ â ¥â +â 2 [Nâ =â 905; 12.6%]). Riskâ adjusted mortality hazard ratios (AHR) were calculated using comprehensive propensity score adjustment by Cox regression.ResultsIncomplete revascularization was rare (4.5%) but associated with increased mortality in all patients (AHR [95% confidence interval]â =â 1.53 [1.29â 1.80]), those undergoing singleâ arterial CABG (AHRâ =â 1.27 [1.04â 1.54]) and multiâ arterial CABG (AHRâ =â 2.18 [1.60â 2.99]), as well as in patients with 3â Vessel (AHRâ =â 1.37 [1.16â 1.62]) and, to a lesser degree, with 2â Vessel (AHRâ =â 1.67 [0.53â 5.23]) coronary disease. Supraâ complete revascularization was generally associated with incrementally decreased mortality in all patients (AHR [CRIâ =â +1]â =â 0.94 [0.87â 1.03]); AHR [CRIâ â ¥â +2]â =â 0.74 [0.64â 0.85]), and was driven by a significantly decreased mortality risk in singleâ arterial CABG (AHR [CRIâ =â +1]â =â 0.90 [0.81â 0.99]; AHR [CRIâ â ¥â +2]â =â 0.64 [0.53â 0.78]); and 3â Vessel disease patients (AHR [CRIâ =â +1]â =â 0.94 [0.86â 1.04]; and AHR [CRIâ â ¥â +2]â =â 0.75 [0.63â 0.88]) with no impact in multiâ arterial CABG (AHR [CRIâ =â +1]â =â 1.07 [0.91â 1.26]; AHR [CRIâ â ¥â +2]â =â 0.93 [0.73â 1.17]).ConclusionsIncomplete revascularization is associated with decreased late survival, irrespective of grafting strategy. Alternatively, supraâ complete revascularization is associated with improved survival in patients with 3â Vessel CAD, and in singleâ arterial but not multiâ arterial CABG.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/146364/1/jocs13810.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/146364/2/jocs13810_am.pd

Topology optimization of nonlinear periodically microstructured materials for tailored homogenized constitutive properties

A topology optimization method is presented for the design of periodic microstructured materials with prescribed homogenized nonlinear constitutive properties over finite strain ranges. The mechanical model assumes linear elastic isotropic materials, geometric nonlinearity at finite strain, and a quasi-static response. The optimization problem is solved by a nonlinear programming method and the sensitivities computed via the adjoint method. Two-dimensional structures identified using this optimization method are additively manufactured and their uniaxial tensile strain response compared with the numerically predicted behavior. The optimization approach herein enables the design and development of lattice-like materials with prescribed nonlinear effective properties, for use in myriad potential applications, ranging from stress wave and vibration mitigation to soft robotics

Acute Hypersensitivity of Pluripotent Testicular Cancer-Derived Embryonal Carcinoma to Low-Dose 5-Aza Deoxycytidine Is Associated with Global DNA Damage-Associated p53 Activation, Anti-Pluripotency and DNA Demethylation

Human embryonal carcinoma (EC) cells are the stem cells of nonseminoma testicular germ cells tumors (TGCTs) and share remarkable similarities to human embryonic stem (ES) cells. In prior work we found that EC cells are hypersensitive to low nanomolar doses of 5-aza deoxycytidine (5-aza) and that this hypersensitivity partially depended on unusually high levels of the DNA methyltransferase, DNMT3B. We show here that low-dose 5-aza treatment results in DNA damage and induction of p53 in NT2/D1 cells. In addition, low-dose 5-aza results in global and gene specific promoter DNA hypomethylation. Low- dose 5-aza induces a p53 transcriptional signature distinct from that induced with cisplatin in NT2/D1 cells and also uniquely downregulates genes associated with pluripotency including NANOG, SOX2, GDF3 and Myc target genes. Changes in the p53 and pluripotency signatures with 5-aza were to a large extent dependent on high levels of DNMT3B. In contrast to the majority of p53 target genes upregulated by 5-aza that did not show DNA hypomethylation, several other genes induced with 5-aza had corresponding decreases in promoter methylation. These genes include RIN1, SOX15, GPER, and TLR4 and are novel candidate tumors suppressors in TGCTs. Our studies suggest that the hypersensitivity of NT2/D1 cells to low-dose 5-aza is multifactorial and involves the combined activation of p53 targets, repression of pluripotency genes, and activation of genes repressed by DNA methylation. Low-dose 5-aza therapy may be a general strategy to treat those tumors that are sustained by cells with embryonic stem-like properties. GEO number for the microarray data: GSE42647

Combining Different Approaches for Grape Pomace Valorization: Polyphenols Extraction and Composting of the Exhausted Biomass

Grape pomace represents 60%, by weight, of the solid side-streams of the wine-making process. The quantities produced, seasonality, and the presence of polyphenols pose economic and environmental issues that require proper management approaches based on the principles of sustainability and circular economy. The present work focuses on the combined application of solid-liquid extraction of polyphenols from ground grape pomace using a hydroethanolic mixture and the composting of the exhausted pomace. The obtained results support the possibility of recovering approximately 76.5 g of extract per kg of dry grape pomace (or 1.8 g of total phenols per kg of dry grape pomace). The composting process was not affected by the extraction process. On the contrary, the composting process was enhanced by the pomace particle size reduction, in terms of final biostability and content of humic acids