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Emad Kordbacheh, Shahram Nazarian, Davoud Sadeghi, Abbas Hajizadeh
Objectives: Enterotoxigenic Escherichia coli (ETEC) is known as the most common bacterial causes of diarrheal diseases related to morbidity and mortality. Heat-labile enterotoxin (LT) is a part of major virulence factors in ETEC pathogenesis. Antigen entrapment into nanoparticles (NPs) can protect them and enhance their immunogenicity. Materials and Methods: In the present study, recombinant LTB protein was expressed in E. coli BL21 (DE3) and purified by an Ni-NTA agarose column...
May 2018: Iranian Journal of Basic Medical Sciences
Chunyan Liu, Gen Zou, Xing Yan, Xuguo Zhou
Termite gut microbiome is a rich reservoir for glycoside hydrolases, a suite of enzymes critical for the degradation of lignocellulosic biomass. To search for hemicellulases, we screened 12,000 clones from a fosmid gut library of a higher termite, Globitermes brachycerastes. As a common Southeastern Asian genus, Globitermes distributes predominantly in tropical rain forests and relies on the lignocellulases from themselves and bacterial symbionts to digest wood. In total, 22 positive clones with β-xylosidase activity were isolated, in which 11 representing different restriction fragment length polymorphism (RFLP) patterns were pooled and subjected to 454 pyrosequencing...
2018: International Journal of Biological Sciences
Weicheng Huang, Jinxin Wang, Lang Bian, Chaoyue Zhao, Danqing Liu, Chongshen Guo, Bin Yang, Wenwu Cao
A WO3-x/TiO2-x nanotube array (NTA) heterojunction photoanode was strategically designed to improve photoelectrocatalytic (PEC) performance by establishing a synergistic vacancy-induced self-doping effect and localized surface plasmon resonance (LSPR) effect of metalloid non-stoichiometric tungsten suboxide. The WO3-x/TiO2-x NTA heterojunction photoanode was synthesized through a successive process of anodic oxidation to form TiO2 nanotube arrays, magnetron sputtering to deposit metalloid WO3-x, and post-hydrogen reduction to engender oxygen vacancy in TiO2-x as well as crystallization...
June 14, 2018: Physical Chemistry Chemical Physics: PCCP
Marco P Monopoli, Andrea Zendrini, Dan Wu, Shane Cheung, Gonzalo Sampedro, Brendan Ffrench, John Nolan, Olga Piskareva, Raymond L Stalings, Serena Ducoli, Paolo Bergese, Donal F O'Shea
The recognition of the biological, diagnostic and medical importance of exosomes has given rise to an urgent need for efficient labelling of these extracellular vesicles in ways that do not alter their inherent characteristics. We report for the first time an endogenous method to NIR-fluorescent labelled exosomes using an amphiphilic probe without the need for immunolabelling or synthetic or chromatographic manipulation of exosomes. Comparative analyses of labelled and unlabelled exosomes with NTA, AFM, flow cytometry and immunoblot analysis all show a high degree of similarity...
June 14, 2018: Chemical Communications: Chem Comm
János András Mótyán, Márió Miczi, Beáta Bozóki, József Tőzsér
Data provided here are related to the research article entitled as 'A recombinant fusion protein-based, fluorescent protease assay for high throughput-compatible substrate screening'. Here we describe data related to the investigation of the properties of the His6 -MBP-VSQNY↓PIVQ-mApple recombinant protein substrate and its interactions with Ni-NTA magnetic beads, including the dependence of substrate attachment on incubation time and concentration. Data on the folding efficiency and conformational stability of the recombinant substrate assessed by tryptic digestion are also presented...
June 2018: Data in Brief
Dujun Wang, Li Zhao, Jiayi Jiang, Jia Liu, Dan Wang, Xiaofeng Yu, Yuan Wei, Zhen Ouyang
1-Deoxynojirimycin (DNJ) is the main bioactive compound of Morus alba L.. DNJ has pharmacological effects, including blood sugar level regulation and antiviral activity. In this study, the mulberry lysine decarboxylase gene (MaLDC), which is involved in the biosynthesis of DNJ alkaloids, was cloned, expressed, and functionally verified. MaLDC was induced and expressed in Escherichia coli BL21 (DE3). The recombinant soluble MaLDC protein had a relative molecular mass of 24.0 kDa. The protein was purified by Ni-NTA separation...
June 9, 2018: Protein Expression and Purification
Anupriya Baranwal, Adarsh Kumar Chiranjivi, Ashutosh Kumar, Vikash Kumar Dubey, Pranjal Chandra
Nanotherapeutic agents (NTA) play a crucial role in clinical medicine, if their unique properties are well understood and well exploited. In this direction, we report synthesis and characterization of highly potent phytofabricated silver nanoparticles (AgNPs) using Sechium edule, which served the purpose of both reducing and capping agent. The designed AgNPs were characterized using UV-Vis spectroscopy, XRD, FTIR, HR-TEM, and TGA techniques. The formation of AgNPs was also confirmed using electrochemistry, which to the best of our knowledge has never been reported before for biosynthesized nanoparticles...
June 11, 2018: Scientific Reports
Fengjie Liu, Claude Fortin, Peter G C Campbell
In this study we tested the hypothesis that metal uptake by unicellular algae may be affected by changes in metal speciation in the boundary layer surrounding the algal cells. The freshwater alga Chlamydomonas reinhardtii was pre-acclimated to different N nutrition regimes; changes in N nutrition are known to change the nature of extracellular metabolites (e.g., reactive oxygen species 'ROS', and OH- ) and thus boundary layer chemical conditions. Specifically, at a constant bulk free Cd2+ concentration, Cd uptake by N-starved algae in cysteine-buffered solution was significantly higher than that in NTA-buffered solution...
June 8, 2018: Environmental Science & Technology
Elisabetta Pancani, Mario Menendez-Miranda, Alexandra Pastor, François Brisset, Marie-Françoise Bernet-Camard, Didier Desmaële, Ruxandra Gref
Nowadays, biodegradable polymers such as poly(lactic acid) (PLA), poly(D,L-lactic- co -glycolic acid) (PLGA) and poly( ε -caprolactone) (PCL) remain the most common biomaterials to produce drug-loaded nanoparticles (NPs). Pipemidic acid (PIP) is a poorly soluble antibiotic with a strong tendency to crystallize. PIP incorporation in PLA/PLGA NPs was challenging because of PIP crystals formation and burst release. As PIP had a poor affinity for the NPs, an alternative approach to encapsulation was used, consisting in coupling PIP to PCL...
May 2018: Acta Pharmaceutica Sinica. B
M Duroux, H S H Vinterhøj, A Stensballe, P Gazerani
Aims Satellite glial cells (SGCs) are non-neuronal cells that entirely surround neurons within sensory ganglia. This unique structure allows SGC-neuron interactions. Altered cross-talk following nerve injury or inflammation is thought to contribute to pathogenesis of chronic pain. Release of extracellular vesicles in form of exosomes has been found to play a key role in cell-cell communication. However, release of exosomes from SGCs and their potential role in modulating pain remain unknown. Hence, this study aimed at identifying and characterizing shed exosomes from SGCs under normal and inflammatory conditions...
December 29, 2017: Scandinavian Journal of Pain
A Sammarco, G Finesso, L Cavicchioli, S Ferro, F Caicci, R Zanetti, R Sacchetto, V Zappulli
Extracellular vesicles (EVs) are membrane-bound vesicles produced by cells, known to play a key role in cell-to-cell communication. They exert pleiotropic biological functions via the horizontal transfer of bioactive molecules (DNA, RNAs, proteins, and lipids) within the tumour microenvironment and throughout the body. In human cancer, EVs are known to interfere with pathways that lead to tumour progression and are used as novel cancer biomarkers. In veterinary medicine, very little is known on cancer-derived EVs...
May 30, 2018: Veterinary and Comparative Oncology
Karl Gatterdam, Eike F Joest, Volker Gatterdam, Robert Tampé
Small chemical/biological interaction pairs are at the forefront in tracing proteins' function and interaction at high signal-to-background ratio in cellular pathways. Pharma ventures have eager plans to develop trisNTA probes for in vitro and in vivo screening of His-tagged protein targets. However, the optimal design of scaffold, linker, and chelator head yet deserves systematic investigations to achieve highest affinity and kinetic stability for in vitro and especially cell applications. In this study, we report on a library of N-nitrilotriacetic acid (NTA) based multivalent chelator heads (MCHs) built up on linear, cyclic, and dendritic scaffolds and contrast these with regard to their binding affinity and stability for labeling of cellular His-tagged proteins...
May 29, 2018: Angewandte Chemie
Paula Martins, Daisy Machado, Thais Holtz Theizen, João Paulo Oliveira Guarnieri, Bruno Gaia Bernardes, Gabriel Piccirillo Gomide, Marcus Alexandre Finzi Corat, Camilla Abbehausen, José Luiz Proença Módena, Carlos Fernando Odir Rodrigues Melo, Karen Noda Morishita, Rodrigo Ramos Catharino, Clarice Weis Arns, Marcelo Lancellotti
The increase of Zika virus (ZIKV) infections in Brazil in the last two years leaves a prophylactic measures on alert for this new and emerging pathogen. Concerning of our positive experience, we developed a new prototype using Neisseria meningitidis outer membrane vesicles (OMV) on ZIKV cell growth in a fusion of OMV in the envelope of virus particles. The fusion of nanoparticles resulting from outer membrane vesicles of N. meningitidis with infected C6/36 cells line were analyzed by Nano tracking analysis (NTA), zeta potential, differential light scattering (DLS), scan and scanning transmission eletronic microscopy (SEM and STEM) and high resolution mass spectometry (HRMS) for nanostructure characterization...
May 29, 2018: Scientific Reports
Yiyi Yang, Antonio Boza-Serrano, Christopher J R Dunning, Bettina Hjelm Clausen, Kate Lykke Lambertsen, Tomas Deierborg
BACKGROUND: Activated microglia play an essential role in inflammatory responses elicited in the central nervous system (CNS). Microglia-derived extracellular vesicles (EVs) are suggested to be involved in propagation of inflammatory signals and in the modulation of cell-to-cell communication. However, there is a lack of knowledge on the regulation of EVs and how this in turn facilitates the communication between cells in the brain. Here, we characterized microglial EVs under inflammatory conditions and investigated the effects of inflammation on the EV size, quantity, and protein content...
May 28, 2018: Journal of Neuroinflammation
Julian Bender, Sabine Bognar, Maurizio Camagna, Julia A M Donauer, Julian W Eble, Ramona Emig, Sabrina Fischer, Rabea Jesser, Luisa Keilholz, Daniel M U Kokotek, Julika Neumann, Simon Nicklaus, Ricardo R Q P T Oude Weernink, Lara G Stühn, Nathalie Wössner, Stefan D Krämer, Philipp Schwenk, Nicole Gensch, Günter Roth, Maximilian H Ulbrich
The detection of antibodies from blood sera is crucial for diagnostic purposes. Miniaturized protein assays in combination with microfluidic setups hold great potential by enabling automated handling and multiplexed analyses. Yet, the separate expression, purification, and storage of many individual proteins are time consuming and limit applicability. In vitro cell-free expression has been proposed as an alternative procedure for the generation of protein assays. We report the successful in vitro expression of different model proteins from DNA templates with an optimized expression mix...
May 12, 2018: Biosensors & Bioelectronics
Yuanjun Zhao, Sarah R Weber, Joshua Lease, Mariano Russo, Christopher A Siedlecki, Li-Chong Xu, Han Chen, Weiwei Wang, Michael Ford, Rafael Simó, Jeffrey M Sundstrom
Purpose: To investigate the molecular components of the vitreous in order to better understand retinal physiology and disease. Methods: Vitreous was acquired from patients undergoing vitrectomy for macular hole and/or epiretinal membrane, postmortem donors, and C57BL/6J mice. Unbiased proteomic analysis was performed via electrospray ionization tandem mass spectrometry (MS/MS). Gene ontology analysis was performed and results were confirmed with transmission electron microscopy, atomic force microscopy, and nanoparticle tracking analysis (NTA)...
May 2018: Translational Vision Science & Technology
Huihui Huang, Zheyan Wang, Zhenggen Yang, Zhen Li, Ping Wang, Wenqi Dong
Objective To construct eukaryotic expression vectors of human IgE heavy chain 2-4 region (IgECepsilon2-4) and purify the recombinant protein, and then capture its interacted proteins by surface plasmon resonance (SPR). Methods Three recombinant eukaryotic expression vectors of IgECepsilon2-4 containing different signal peptides were constructed and transiently transfected into HEK293FT suspension cells separately. The recombinant plasmid with the highest-level expression was selected to express the recombinant protein in a huge amount, and then the recombinant protein was purified by Ni-NTA affinity chromatography...
March 2018: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Yousef Amini, Mohsen Tafaghodi, Saeid Amel Jamehdar, Zahra Meshkat, Bagher Moradi, Mojtaba Sankian
Background: Subunit vaccines are appropriate vaccine candidates for the prevention of some infections. In this study, three immunogenic proteins of Mycobacterium tuberculosis , including HspX, Ppe44, and EsxV as a new construction, were expressed alone and as a fusion protein to develop a new vaccine candidate against tuberculosis infection. Methods: To make the fusion protein, the three genes were linked together by AEAAAKEAAAKA linkers and inserted into pET21b and pET32b vectors...
April 2018: Reports of Biochemistry & Molecular Biology
Ge Ge Sun, Yan Yan Song, Peng Jiang, Hua Na Ren, Shu Wei Yan, Yue Han, Ruo Dan Liu, Xi Zhang, Zhong Quan Wang, Jing Cui
BACKGROUND: Trichinellosis is a serious zoonositc parasitosis worldwide. Because its clinical manifestations aren't specific, the diagnosis of trichinellosis is not easy to be made. Trichinella spiralis muscle larva (ML) excretory-secretory (ES) antigens are the most widely applied diagnostic antigens for human trichinellosis, but the major drawback of the ES antigens for assaying anti-Trichinella antibodies is the false negative in the early Trichinella infection period. The aim of this study was to characterize the T...
May 14, 2018: PLoS Neglected Tropical Diseases
Yang Yang, Li Cheng Kao, Yuanyue Liu, Ke Sun, Hongtao Yu, Jinghua Guo, Sofia Ya Hsuan Liou, Michael R Hoffmann
TiO2 has long been recognized as a stable and reusable photocatalyst for water splitting and pollution control. However, it is an inefficient anode material in the absence of photoactivation due to its low electron conductivity. To overcome this limitation, a series of conductive TiO2 nanotube array electrodes have been developed. Even though nanotube arrays are effective for electrochemical oxidation initially, deactivation is often observed within a few hours. To overcome the problem of deactivation, we have synthesized cobalt-doped Black-TiO2 nanotube array (Co-Black NTA) electrodes that are stable for more than 200 h of continuous operation in a NaClO4 electrolyte at 10 mA cm-2 ...
May 4, 2018: ACS Catalysis
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