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

Establishment and characterization of two human breast carcinoma cell lines by spontaneous immortalization: Discordance between Estrogen, Progesterone and HER2/neu receptors of breast carcinoma tissues with derived cell lines

Background: Breast cancer is one of the most common cancers among women throughout the world. Therefore, established cell lines are widely used as in vitro experimental models in cancer research.Methods: Two continuous human breast cell lines, designated MBC1 and MBC2, were successfully established and characterized from invasive ductal breast carcinoma tissues of Malaysian patients. MBC1 and MBC2 have been characterized in terms of morphology analysis, population doubling time, clonogenic formation, wound healing assay, invasion assay, cell cycle, DNA profiling, fluorescence immunocytochemistry, Western blotting and karyotyping.Results: MBC1 and MBC2 exhibited adherent monolayer epithelial morphology at a passage number of 150. Receptor status of MBC1 and MBC2 show (ER+, PR+, HER2+) and (ER+, PR-, HER2+), respectively. These results are in discordance with histopathological studies of the tumoral tissues, which were triple negative and (ER-, PR-, HER2+) for MBC1 and MBC2, respectively. Both cell lines were capable of growing in soft agar culture, which suggests their metastatic potential. The MBC1 and MBC2 metaphase spreads showed an abnormal karyotype, including hyperdiploidy and complex rearrangements with modes of 52-58 chromosomes per cell.Conclusions: Loss or gain in secondary properties, deregulation and specific genetic changes possibly conferred receptor changes during the culturing of tumoral cells. Thus, we hypothesize that, among heterogenous tumoral cells, only a small minority of ER+/PR+/HER2+ and ER+/PR-/HER2+ cells with lower energy metabolism might survive and adjust easily to in vitro conditions. These cell lines will pave the way for new perspectives in genetic and biological investigations, drug resistance and chemotherapy studies, and would serve as prototype models in Malaysian breast carcinogenesis investigations. © 2012 Kamalidehghan et al.; licensee BioMed Central Ltd

Stir casting process for manufacture of Al–SiC composites

Stir casting is an economical process for the fabrication of aluminum matrix composites. There are many parameters in this process, which affect the final microstructure and mechanical properties of the composites. In this study, micron-sized SiC particles were used as reinforcement to fabricate Al-3 wt% SiC composites at two casting temperatures (680 and 850 C) and stirring periods (2 and 6 min). Factors of reaction at matrix/ceramic interface, porosity, ceramic incorporation, and agglomeration of the particles were evaluated by scanning electron microscope (SEM) and high-resolution transition electron microscope (HRTEM) studies. From microstructural characterizations, it is concluded that the shorter stirring period is required for ceramic incorporation to achieve metal/ceramic bonding at the interface. The higher stirring temperature (850 C) also leads to improved ceramic incorporation. In some cases, shrinkage porosity and intensive formation of Al4C3 at the metal/ceramic interface are also observed. Finally, the mechanical properties of the composites were evaluated, and their relation with the corresponding microstructure and processing parameters of the composites was discussed

Bacterial iron-oxide nanowires from biofilm waste as a new adsorbent for the removal of arsenic from water

Biofilm, generated by the bacteria in the groundwater pumping system pipelines of the Salt Interception Scheme on the River Murray in South Australia is discarded as a waste material accumulated after periodic cleaning of the pipes. Structural and chemical composition characterizations confirm that this waste material is composed of amorphous twisted iron-oxide nanowires (ION), generated by bacteria, and they have a unique structure and properties. The adsorption performance of these iron-oxide nanowires for arsenic removal from water was evaluated to define their adsorption capacity for As(III) and As(V) and kinetics. Obtained results demonstrate considerable adsorption properties of this waste biological material and suggest its promising application as a new and low-cost adsorbent for water treatment.Ivan Andjelkovic, Sara Azari, Mason Erkelens, Peter Forward, Martin F. Lambert and Dusan Losi

Gambaran Skala Nyeri Pada Anak Dengan Menggunakan Skala Nyeri Flacc Scale Saat Tindakan Invasif

Face, Legs, Activity, Cry, and Consolability (FLACC) scale is a measurement of pain which already has good validity. This scale can be used for children aged 2 months to 7 years old. There are five points that were observed on this scale. The points were facial, foot activity, crying rate, and the capability to be entertained. Each point has a rating of 0-2 with a maximum value of 10. Response of no pain (0), mild pain (1-3), moderate pain (4-6), and severe pain (7-10) so that the scale can be used to assess the scale of pain in children. This research was conducted during the May to June 2015. The aim of this research was to reveal children\u27s pain scale using FLACC pain scale when invasive procedure in RSUD Arifin Ahcmad was conducted. This research used simple descriptive design. 30 respondents were taken with consecutive sampling technique. The result showed that there was mild pain response in 3 respondents (21,4%), moderate pain response in 7 respondents (50,0%), and severe pain response in 4 respondents (28,6%). According to the resear cher this study recommended that nurses to use FLACC scale to identify the pain scale in children from aged 2 month to 7 years old. so the need consolability can be done as soon as possible

Fabrication of aluminum matrix composites reinforced with nano- to micrometer-sized SiC particles

In this study, the hot extrusion process was applied to stir cast aluminum matrix–SiC composites in order to improve their microstructure and reduce cast part defects. SiC particles were ball milled with Cr, Cu, and Ti as three forms of carrier agents to improve SiC incorporation. Large brittle ceramic particles (average particle size: 80 μm) were fragmented during ball-milling to form nanoparticles in order to reduce the cost of composite manufacturing. The experimental results indicate that full conversion of coarse micron sized to nanoparticles, even after 36 h of ball milling, was not possible. Multi modal SiC particle size distributions which included SiC nanoparticles were produced after the milling process, leading to the incorporation of a size range of SiC particle sizes from about 50 nm to larger than 10 μm, into the molten A356 aluminum alloy. The particle size of the milled powders and the amount of released heat from the reaction between the carrier agent and molten aluminum are inferred as two crucial factors that affect the resultant part tensile properties and microhardness

Catalytic decomposition of 2-chlorophenol using an ultrasonic-assisted Fe3O4-TiO2@MWCNT system: Influence factors, pathway and mechanism study.

As a reusable sonocatalyst, magnetically separable Fe3O4-TiO2@MWCNT (FMT) was synthesized by an ultrasound-assisted wet impregnation method and was evaluated in the removal of 2-chlorophenol (2CP). Physical and chemical properties of the catalyst composite materials were investigated by all catalysts were systematically characterized using Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), X-ray Photoelectron Spectroscopy (XPS), Energy Dispersive X-ray Analysis (EDX), Dynamic light scattering (DLS), and N2-physisorption. The efficiency and kinetics of 2CP removal by FMT-assisted sonocatalysis (FMT-US) was systematically investigated under various operational parameters i.e. pH, FMT and 2CP concentration, temperature and ultrasonic power. The results indicated that 0.4gL-1 FMT dosage, pH 5, temperature of 35°C as well as 50 w ultrasound power are the most favorable conditions for the degradation of the 2CP. Furthermore, both of the superoxide and hydroxyl radicals were produced in the reaction, however, superoxide radicals were assumed to be the dominating reactive species for the 2CP degradation, according to the scavenging tests and electron paramagnetic resonance tests. Moreover, the FMT catalyst exhibited a high reusability and stability in the US/FMT system during the five repetitive experiments. The intermediate products were identified by GC-MS, thereby a possible degradation pathway is proposed. The chemical oxygen demand (COD) and corresponding total organic carbon (TOC) removal efficiencies were 64.9% and 56.7%, respectively. Finally, toxicity tests showed that the toxicity of the solution increased during the first 5min and then decreased significantly with the progress of the oxidation. The mechanisms of ultrasound irritation enhanced FMT activation were also proposed. Copyright © 2017 Elsevier Inc. All rights reserved

Multi-walled carbon nanotubes modified with iron oxide and silver nanoparticles (MWCNT-Fe<inf>3</inf>O<inf>4</inf>/Ag) as a novel adsorbent for determining PAEs in carbonated soft drinks using magnetic SPE-GC/MS method

The synthesis of compounds with an excellent adsorption capability plays an essential role to remove contaminants such as phthalic acid esters (PAEs) with potential carcinogenic characteristics from different food products. In this context, for the first time, a novel adsorbent (MWCNT-Fe3O4/Ag) was synthesized by using iron (magnetic agent), and silver (catalytic and surface enhancer agent) to further approach in a magnetic SPE-GC/MS method for determining of PAEs in carbonated soft drink samples. The limit of detection (LOD) and limit of quantification (LOQ) values of MSPE-GC/MS were determined in six PAEs as a range of 10.8–22.5 and 36–75 ng/L, respectively. Also, the calibration curves of PAEs were linear (R2 = 0.9981–0.9995) over the concentration level of 10.000 ng/L and the recoveries of the six PAEs were ranging from 96.60% to 109.22% with the RSDs less than 8%. Moreover, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), X-ray diffraction (XRD), vibrating sample magnetometry (VSM) and transmission electron microscopy analyses (TEM) were utilized to characterize the produced MWCNT-Fe3O4/Ag. Based on the findings, the surface of MWCNT is relatively uniform, which became coarser after loading with Fe3O4/Ag particles. Also, EDX spectrum showed the carbon (C), iron (Fe), oxygen (O), Ag and copper (Cu) are the main components of synthesized MWCNTs-Fe3O4/Ag. The successful adhesion of Fe3O4/Ag on the texture of MWCNTs using a co-precipitation method was confirmed by XRD and FT-IR assays. Additionally, excellent crystallinity and clear lattice nanocrystals fringes of prepared MWCNT-Fe3O4/Ag was demonstrated by TEM analysis. Based on the obtained VSM images, the prepared sorbent (MWCNT-Fe3O4/Ag) has the good magnetic performance for magnetic separation and extraction processes. It was concluded that the synthesized MWCNT-Fe3O4/Ag could be used as an efficient adsorbent for determining contaminants such as PAEs in different beverage samples. © 201

A Survey of Air-to-Ground Propagation Channel Modeling for Unmanned Aerial Vehicles

In recent years, there has been a dramatic increase in the use of unmanned aerial vehicles (UAVs), particularly for small UAVs, due to their affordable prices, ease of availability, and ease of operability. Existing and future applications of UAVs include remote surveillance and monitoring, relief operations, package delivery, and communication backhaul infrastructure. Additionally, UAVs are envisioned as an important component of 5G wireless technology and beyond. The unique application scenarios for UAVs necessitate accurate air-to-ground (AG) propagation channel models for designing and evaluating UAV communication links for control/non-payload as well as payload data transmissions. These AG propagation models have not been investigated in detail when compared to terrestrial propagation models. In this paper, a comprehensive survey is provided on available AG channel measurement campaigns, large and small scale fading channel models, their limitations, and future research directions for UAV communication scenarios

Cellular connectivity for UAVs: Network modeling, performance analysis, and design guidelines

The growing use of aerial user equipments (UEs) in various applications requires ubiquitous and reliable connectivity for safe control and data exchange between these devices and ground stations. Key questions that need to be addressed when planning the deployment of aerial UEs are whether the cellular network is a suitable candidate for enabling such connectivity and how the inclusion of aerial UEs might impact the overall network efficiency. This paper provides an in-depth analysis of user and network-level performance of a cellular network that serves both unmanned aerial vehicles (UAVs) and ground users in the downlink. Our results show that the favorable propagation conditions that UAVs enjoy due to their height often backfire on them, as the increased load-dependent co-channel interference received from neighboring ground base stations (BSs) is not compensated by the improved signal strength. When compared with a ground user in an urban area, our analysis shows that a UAV flying at 100 m can experience a throughput decrease of a factor 10 and a coverage drop from 76% to 30%. Motivated by these findings, we develop UAV and network-based solutions to enable an adequate integration of UAVs into cellular networks. In particular, we show that an optimal tilting of the UAV antenna can increase the coverage from 23% to 89% and throughput from 3.5 to 5.8 b/s/Hz, outperforming ground UEs. Furthermore, our findings reveal that depending on the UAV altitude and its antenna configuration, the aerial user performance can scale with respect to the network density better than that of a ground user. Finally, our results show that network densification and the use of microcells limit the UAV performance. Although UAV usage has the potential to increase the area spectral efficiency (ASE) of cellular networks with a moderate number of cells, they might hamper the development of future ultradense networks