RESUMEN
Rapid and accurate detection of lethal volatile compounds is an emerging requirement to ensure the security of the current and future society. Since the threats are becoming more complex, the assurance of future sensing devices' performance can be obtained solely based on a thorough fundamental approach, by utilizing physics and chemistry together. In this work, we have applied thermal desorption spectroscopy (TDS) to study dimethyl methylophosphate (DMMP, sarin analogue) adsorption on zinc phthalocyanine (ZnPc), aiming to achieve the quantification of the sensing mechanism. Furthermore, we utilize a novel approach to TDS that involves quantum chemistry calculations for the determination of desorption activation energies. As a result, we have provided a comprehensive description of DMMP desorption processes from ZnPc, which is the basis for successful future applications of sarin ZnPc-based sensors. Finally, we have verified the sensing capability of the studied material at room temperature using impedance spectroscopy and took the final steps towards demonstrating ZnPc as a promising sarin sensor candidate.
Asunto(s)
Sustancias para la Guerra Química , Compuestos Organometálicos , Sustancias para la Guerra Química/análisis , Sarín , Compuestos Organometálicos/química , Compuestos de ZincRESUMEN
Due to its position at the outermost of glycans, sialic acid is involved in a myriad of physiological and pathophysiological cell functions such as host-pathogen interactions, immune regulation, and tumor evasion. Inhibitors of cell surface sialylation could be a useful tool in cancer, immune, antibiotic, or antiviral therapy. In this work, four different C-3 modified N-acetylmannosamine analogs were tested as potential inhibitors of cell surface sialylation. Peracetylated 2-acetylamino-2-deoxy-3-O-methyl-D-mannose decreases cell surface sialylation in Jurkat cells in a dose-dependent manner up to 80%, quantified by flow cytometry and enzyme-linked lectin assays. High-performance liquid chromatography experiments revealed that not only the concentration of membrane bound but also of cytosolic sialic acid is reduced in treated cells. We have strong evidence that the observed reduction of sialic acid expression in cells is caused by the inhibition of the bifunctional enzyme UDP-GlcNAc-2-epimerase/ManNAc kinase. 2-Acetylamino-2-deoxy-3-O-methyl-D-mannose inhibits the human ManNAc kinase domain of the UDP-GlcNAc-2-epimerase/ManNAc kinase. Binding kinetics of the inhibitor and human N-acetylmannosamine kinase were evaluated using surface plasmon resonance. Specificity studies with human N-acetylglucosamine kinase and hexokinase IV indicated a high specificity of 2-acetylamino-2-deoxy-3-O-methyl-D-mannose for MNK. This substance represents a novel class of inhibitors of sialic acid expression in cells, targeting the key enzyme of sialic acid de novo biosynthesis.
Asunto(s)
Carbohidrato Epimerasas/química , Proteínas Portadoras/química , Hexosaminas/química , Ácido N-Acetilneuramínico/química , Cromatografía Líquida de Alta Presión , Clonación Molecular , Citosol/metabolismo , Relación Dosis-Respuesta a Droga , Citometría de Flujo , Humanos , Células Jurkat , Cinética , Lectinas , Fosfotransferasas (Aceptor de Grupo Alcohol)/química , Unión Proteica , Dispersión de Radiación , Especificidad por Sustrato , Resonancia por Plasmón de SuperficieRESUMEN
Multimeric glycoprotein von Willebrand factor (VWF) exhibits a unique triplet structure of individual oligomers, resulting from ADAMTS-13 (a disintegrin and metalloproteinase with thrombospondin type 1 motifs 13) cleavage. The faster and slower migrating triplet bands of a given VWF multimer have one shorter or longer N-terminal peptide sequence, respectively. Within this peptide sequence, the A1 domain regulates interaction of VWF with platelet glycoprotein (GP)Ib. Therefore, platelet-adhesive properties of two VWF preparations with similar multimeric distribution but different triplet composition were investigated for differential functional activities. Preparation A was enriched in intermediate triplet bands, whereas preparation B predominantly contained larger triplet bands. Binding studies revealed that preparation A displayed a reduced affinity for recombinant GPIb but an unchanged affinity for collagen type III when compared to preparation B. Under high-shear flow conditions, preparation A was less active in recruiting platelets to collagen type III. Furthermore, when added to blood from patients with von Willebrand disease (VWD), defective thrombus formation was less restored. Thus, VWF forms lacking larger-size triplet bands appear to have a decreased potential to recruit platelets to collagen-bound VWF under arterial flow conditions. By implication, changes in triplet band distribution observed in patients with VWD may result in altered platelet adhesion at high-shear flow.
Asunto(s)
Plaquetas/metabolismo , Complejo GPIb-IX de Glicoproteína Plaquetaria/metabolismo , Trombosis/sangre , Factor de von Willebrand/metabolismo , Plaquetas/citología , Ensayo de Inmunoadsorción Enzimática , Humanos , Adhesividad Plaquetaria/fisiología , Resonancia por Plasmón de Superficie , Trombosis/patologíaRESUMEN
Background: Applications of two-dimensional (2D) materials in electronic devices require the development of appropriate measuring methods for determining their typical semiconductor parameters, i.e., mobility and carrier lifetime. Among these methods, contactless techniques and mobility extraction methods based on field-effect measurements are of great importance. Results: Here we show a contactless method for determining these parameters in 2D semiconductors that is based on the photomagnetoelectric (PME) effect (also known as the photoelectromagnetic effect). We present calculated dependences of the PME magnetic moment, evoked in 2D Corbino configuration, on the magnetic field as well as on the intensity and spatial distribution of illumination. The theoretical predictions agree with the results of the contactless investigations performed on non-suspended single-layer graphene. We use the contactless PME method for determining the dependence of carrier mobility on the concentration of electrons and holes induced by a back-gate voltage. Conclusion: The presented contactless PME method, used in Corbino geometry, is complementary to the mobility extraction methods based on field-effect measurements. It can be used for determining the mobility and diffusion length of carriers in different 2D materials.
RESUMEN
INTRODUCTION: von Willebrand factor (VWF) is rich in cysteine; next to important structural disulfide bonds, free thiol groups are present. Free thiols on the surface of plasmatic VWF have been shown to play a role in VWF self-association and in platelet binding under pathologically high levels of shear stress. The present study explores the role of VWF free thiol groups under physiological levels of shear stress and in interactions with collagen and platelet-GPIbα receptor. MATERIALS AND METHODS: Free and accessible thiol groups were blocked with N-ethylmaleimide (NEM) and the derivatized molecule was evaluated in functional assays. Reduced cysteine residues were identified using biotin-linked maleimide (MPB) followed by analysis of multimer and domain incorporation and by analysis of derivatized tryptic peptides by mass spectrometry. RESULTS: Blockade of free thiol groups significantly reduced VWF-mediated platelet recruitment to collagen under physiological flow conditions. This resulted from inhibition of VWF binding to both collagen and the platelet GPIb receptor. Evaluation of derivatization sites revealed a high level of derivatization in the cysteine-rich N- and C-termini of VWF. 19 MPB-derivatized peptides, 13 of which are described here for the first time, were identified by mass spectrometry. CONCLUSIONS: This study shows a significant contribution of free thiol groups in VWF to the mediation of platelet adhesion under physiological shear stress conditions. The free thiol groups are shown to be involved in VWF binding to both collagen III and platelet GP1b receptor.