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

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

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

Power allocation in multi-hop OFDM transmission systems with amplify-and-forward relaying: A unified approach

In this paper, a unified approach for power allocation (PA) in multi-hop orthogonal frequency division multiplexing (OFDM) amplify-and-forward (AF) relaying systems is presented. In the proposed approach, we consider short and long term individual and total power constraints at the source and relays, and devise low complexity PA algorithms when wireless links are subject to channel path-loss and small-scale Rayleigh fading. To manage the complexity, in the proposed formulations, we adopt a two-stage iterative approach consisting of a power distribution phase among distinct subcarriers, and a power allocation phase among different relays. In particular, aiming at improving the instantaneous rate of multi-hop transmission systems with AF relaying, we develop (i) a near-optimal iterative PA algorithm based on the exact analysis of the received SNR at the destination; (ii) a low complexity suboptimal iterative PA algorithm based on an approximate expression of the received SNR at high-SNR regime; and (iii) a low complexity non-iterative PA scheme with limited performance loss. Simulation results show the superior performance of the proposed power allocation algorithms

A novel, eco-friendly and green synthesis of PPAC-ZnO and PPAC-nZVI nanocomposite using pomegranate peel: Cephalexin adsorption experiments, mechanisms, isotherms and kinetics

In the present work, powdered activated carbon coated by nanoparticles ZnO and nZVI was derived from pomegranate peel extracts and finally applied for removal of cephalexin (CEX (from aqueous solutions. This experimental research was conducted discontinuously. The effects of pH of solution, reaction time, PPAC-nZVI and PPAC-ZnO composites dose, and initial concentration of cephalexin and composite recovery on process efficiency were investigated. The removal efficiency in optimal conditions for cephalexin with PPAC-nZVI and PPAC-ZnO (CEX = 50 mg L�1, composite dose = 1.25 g L�1, reaction time = 45 min and pH = 5) was obtained 96.06 and 94.17, respectively. The results of the study of isotherm and absorption kinetics for both composites showed that the absorption process follows Langmuir isotherm and pseudo second-order kinetics. The present study showed that the composites could be used as an effective and bio-friendly absorbent to remove cephalexin from aqueous solutions. © 2020 The Society of Powder Technology Japa

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

Organizational culture and the organizational commitment; Correlational study in hospital staffs

Organizations need the committed manpower to provide effectiveness and progress which is an important factor in order to achieve organizational goals. According to this, organizational culture increases employees commitment in the organization by their accepted norms and values. Actually, the organization environment & its prevailing culture, affects on the organizational commitment of staff. This study was conducted to assess relationship between the organizational culture and the organizational commitment among staff of elected hospirals in Tehran University of Medical Sciences. This descriptive study accomplished at 2013. The study population was hospitals affiliated to Tehran University of Medical Sciences included the two hospitals �Vali Asr� & �farabi�. Also in hospitals, study populations were all hospital staffs (Administrative and Clinical). Data collection was done using a three parts questionnaire including: the first one contains background information, the second part consists of questions related to organizational culture based on Denison model and the third part includes questions related to organizational Commitment based on Porter model. Validity and reliability were confirmed through getting the opinions of experts and using Cronbach�s alpha. Data analysis was done by SPSS software using Pearson correlation test and the test �t� to determine the relation between the areas of culture and commitment to the type of hospitals. In organizational culture, maximum average & standard deviation has been seen in professional identity (4.74±1.38). Except for the two dimentions of conflict & communication patterns, a significant relation between the organizational culture dimentions and two hospitalas was seen. Also, in the organizational commitment, the maximum average & standard deviation was seen in continuous commitment scope. Overally, average & standard deviation (3.18±0.76) has been obtained for organizational commitment component; also there was significant relation between the emotional commitment dimention & the type of the hospital. This relationship wasn�t meaningful in another dimentions. Except for the conflict dimension, there was significant correlations between the organizational culture dimension& the organizational commitment (p<0.05). Also, this correlation was between the organizational culture and the organizational commitment (r=0.552) Designing an appropriate and justified "reward mechanism" and development of "teamwork" skills between employees can increase their organizational commitment positively and improve hospital performances. © 2015, Canadian Center of Science and Education. All right reserved