Expression of functional recombinant human tissue transglutaminase (TG2) using the bac-to-bac baculovirus expression system

Purpose: Tissue transglutaminase (TG2) is a unique multifunctional enzyme. The enzyme possesses enzymatic activities such as transamidation/crosslinking and non-enzymatic functions such as cell migration and signal transduction. TG2 has been shown to be involved in molecular mechanisms of cancers and several neurodegenerative diseases such as Alzheimer's disease. The present study aimed at cloning and expression of full length human TG2 in Bac-to-Bac baculovirus expression system and evaluation of its activity. Methods: pFastBac HTA donor vector containing coding sequence of human TG2 was constructed. The construct was transformed to DH10Bac for generating recombinant bacmid. The verified bacmid was transfected to insect cell line (Sf9). Expression of recombinant TG2 was examined by RT-PCR, SDS-PAGE and western blot analysis. Functional analysis was evaluated by fluorometric assay and gel electrophoresis. Results: Recombinant bacmid was verified by amplification of a band near to 4500 bp. Expression analysis showed that the enzyme was expressed as a protein with a molecular weight near 80 kDa. Western blot confirmed the presence of TG2 and the activity assays including flurometric assay indicated that the recombinant TG2 was functional. The electrophoresis assay conformed that the expressed TG2 was the indeed capable of crosslinking in the presence of physiological concentration calcium ions. Conclusion: Human TG2 was expressed efficiently in the active biological form in the Bacto- Bac baculovirus expression system. The expressed enzyme could be used for medical diagnostic, or studies which aim at finding novel inhibitors of the enzymes . To best of our knowledge, this is probably the first report of expression of full length human tissue transglutaminase (TG2) using the Bac-to-Bac expression system. © 2016 The Authors

Hopf-cyclic cohomology of the Connes-Moscovici Hopf algebras with infinite dimensional coefficients

We discuss a new strategy for the computation of the Hopf-cyclic cohomology of the Connes-Moscovici Hopf algebra $\mathcal{H}_n$. More precisely, we introduce a multiplicative structure on the Hopf-cyclic complex of $\mathcal{H}_n$, and we show that the van Est type characteristic homomorphism from the Hopf-cyclic complex of $\mathcal{H}_n$ to the Gelfand-Fuks cohomology of the Lie algebra $W_n$ of formal vector fields on $\mathbb{R}^n$ respects this multiplicative structure. We then illustrate the machinery for $n=1$.Comment: Minor revisions to highlight the main result

Preliminary assessment of various additives on the specific reactivity of anti- rHBsAg monoclonal antibodies

Background: Antibodies have a wide application in diagnosis and treatment. In order to maintain optimal stability of various functional parts of antibodies such as antigen binding sites, several approaches have been suggested. Using additives such as polysaccharides and polyols is one of the main methods in protecting antibodies against aggregation or degradation in the formulation. The aim of this study was to evaluate the protective effect of various additives on the specific reactivity of monoclonal antibodies (mAbs) against recombinant HBsAg (rHBsAg) epitopes. Methods: To estimate the protective effect of different additives on the stability of antibody against conformational epitopes (S3 antibody) and linear epitopes (S7 and S11 antibodies) of rHBsAg, heat shock at 37°C was performed in liquid and solid phases. Environmental factors were considered to be constant. The specific reactivity of antibodies was evaluated using ELISA method. The data were analyzed using SPSS software by Mann-Whitney nonparametric test with the confidence interval of 95%. Results: Our results showed that 0.25 M sucrose, 0.04 M trehalose and 0.5% BSA had the most protective effect on maintaining the reactivity of mAbs (S3) against conformational epitopes of rHBsAg. Results obtained from S7 and S11 mAbs against linear characteristics showed minor differences. The most efficient protective additives were 0.04 M trehalose and 1 M sucrose. Conclusion: Nowadays, application of appropriate additives is important for increasing the stability of antibodies. It was concluded that sucrose, trehalose and BSA have considerable effects on the specific reactivity of anti rHBsAg mAbs during long storage. © 2015, Avicenna Journal of Medical Biotechnology. All rights reserved

Topology- and symmetry-protected domain wall conduction in quantum Hall nematics

We consider domain walls in nematic quantum Hall ferromagnets predicted to form in multivalley semiconductors, recently probed by scanning tunnelling microscopy experiments on Bi(111) surfaces. We show that the domain wall properties depend sensitively on the filling factor $\nu$ of the underlying (integer) quantum Hall states. For $\nu=1$ and in the absence of impurity scattering we argue that the wall hosts a single-channel Luttinger liquid whose gaplessness is a consequence of valley and charge conservation. For $\nu=2$, it supports a two-channel Luttinger liquid, which for sufficiently strong interactions enters a symmetry-preserving thermal metal phase with a charge gap coexisting with gapless neutral intervalley modes. The domain wall physics in this state is identical to that of a bosonic topological insulator protected by $U(1)\times U(1)$ symmetry, and we provide a formal mapping between these problems. We discuss other unusual properties and experimental signatures of these `anomalous' one-dimensional systems.Comment: 11 pages, 3 figures, published versio

Novel Phenomena in Dilute Electron Systems in Two Dimensions

We review recent experiments that provide evidence for a transition to a conducting phase in two dimensions at very low electron densities. The nature of this phase is not understood, and is currently the focus of intense theoretical and experimental attention.Comment: To appear as a Perspective in the Proceedings of the National Academy of Sciences. Reference to Chakravarty, Kivelson, Nayak, and Voelker's paper added (Phil. Mag., in press

Effect of sub-zero cooling on microstructure and mechanical properties of a low alloyed austempered ductile iron

The effect of sub-zero cooling on microstructure and mechanical properties of a low alloyed austempered ductile iron has been investigated. Austempering of samples was performed at 325! and 400℃ after austenitizing at 875℃ and 950℃. The sub-zero treatments were carried out by cooling down the samples to -30℃, -70℃ and -196℃. The changes in volume fraction of austenite and mechanical properties were determined after cooling to each temperature. The austenite volume fraction of samples which were austenitized at 875℃ and austempered at 325℃ remained unchanged, whilst it reduced in samples austenitized at 950℃ and 875℃ for austempering temperature of 400℃. In these specimens, some austenite transformed to martensite after subzero cooling. Mechanical property measurements showed a slight increase in strength and hardness and decrease in elongation and toughness due to this transformation behavior

Mapping the wavefunction of transition metal acceptor states in the GaAs surface

We utilize a single atom substitution technique with spectroscopic imaging in a scanning tunneling microscope (STM) to visualize the anisotropic spatial structure of magnetic and non-magnetic transition metal acceptor states in the GaAs (110) surface. The character of the defect states play a critical role in the properties of the semiconductor, the localization of the states influencing such things as the onset of the metal-insulator transition, and in dilute magnetic semiconductors the mechanism and strength of magnetic interactions that lead to the emergence of ferromagnetism. We study these states in the GaAs surface finding remarkable similarities between the shape of the acceptor state wavefunction for Mn, Fe, Co and Zn dopants, which is determined by the GaAs host and is generally reproduced by tight binding calculations of Mn in bulk GaAs [Tang, J.M. & Flatte, M.E., Phys. Rev. Lett. 92, 047201 (2004)]. The similarities originate from the antibonding nature of the acceptor states that arise from the hybridization of the impurity d-levels with the host. A second deeper in-gap state is also observed for Fe and Co that can be explained by the symmetry breaking of the surface.Comment: 19 pages, 6 figure

Mechanical performance of composite bonded joints in the presence of localised process-induced zero-thickness defects

Processing parameters and environmental conditions can introduce variation into the performance of adhesively bonded joints. The effect of such variation on the mechanical performance of the joints is not well understood. Moreover, there is no validated nondestructive inspection (NDI) available to ensure bond integrity post-process and in-service so as to guarantee initial and continued airworthiness in aerospace sector. This research studies polymer bond defects produced in the laboratory scale single-lap composite-to-composite joints that may represent the process-induced defects occurring in actual processing scenarios such as composite joining and repair in composite aircrafts. The effect of such defects on the degradation of a joint's mechanical performance is then investigated via quasi-static testing in conjunction with NDI ultrasonic C-scanning and pulsed thermography. This research is divided into three main sections: 1- manufacturing carbon fibre-reinforced composite joints containing representative nearly zero-thickness bond defects, 2- mechanical testing of the composite joints, and 3- assessment of the NDI capability for detection of the bond defects in such joints

Electrospun Piezoelectric Polymer Nanofiber Layers for Enabling in Situ Measurement in High-Performance Composite Laminates

This article highlights the effects from composite manufacturing parameters on fiber-reinforced composite laminates modified with layers of piezoelectric thermoplastic nanofibers and a conductive electrode layer. Such modifications have been used for enabling in situ deformation measurement in high-performance aerospace and renewable energy composites. Procedures for manufacturing high-performance composites are well-known and standardized. However, this does not imply that modifications via addition of functional layers (e.g., piezoelectric nanofibers) while following the same manufacturing procedures can lead to a successful multifunctional composite structure (e.g., for enabling in situ measurement). This article challenges success of internal embedment of piezoelectric nanofibers in standard manufacturing of high-performance composites via relying on composite process specifications and parameters only. It highlights that the process parameters must be revised for manufacturing of multifunctional composites. Several methods have been used to lay up and manufacture composites such as electrospinning the thermoplastic nanofibers, processing an inter digital electrode (IDE) made by conductive epoxy-graphene resin, and prepreg autoclave manufacturing aerospace grade laminates. The purpose of fabrication of IDE was to use a resin type (HexFlow RTM6) for the conductive layer similar to that used for the composite. Thereby, material mismatch is avoided and the structural integrity is sustained via mitigation of downgrading effects on the interlaminar properties. X-ray diffraction, Fourier transform infrared spectroscopy, energy dispersive X-ray spectroscopy, and scanning electron microscopy analyses have been carried out in the material characterization phase. Pulsed thermography and ultrasonic C-scanning were used for the localization of conductive resin embedded within the composite laminates. This study also provides recommendations for enabling internally embedded piezoelectricity (and thus health-monitoring capabilities) in high-performance composite laminates

An extension of the spectral Tau method for numerical solution of multi-order fractional differential equations with convergence analysis

AbstractThe main purpose of this paper is to provide an efficient numerical approach for the fractional differential equations (FDEs) based on a spectral Tau method. An extension of the operational approach of the Tau method with the orthogonal polynomial bases is proposed to convert FDEs to its matrix–vector multiplication representation. The fractional derivatives are described in the Caputo sense. The spectral rate of convergence for the proposed method is established in the L2 norm. We tested our procedure on several examples and observed that the obtained numerical results confirm the theoretical prediction of the exponential rate of convergence