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https://www.readbyqxmd.com/read/28811265/molecular-cloning-heterologous-expression-and-functional-characterization-of-gamma-tocopherol-methyl-transferase-%C3%AE-tmt-from-glycine-max
#1
Kalpana Tewari, Anil Dahuja, Archana Sachdev, Kishwar Ali, Vaibhav Kumar, Amresh Kumar, Sweta Kumari
γ-Tocopherol methyltransferase (γ-TMT) (EC 2.1.1.95) is the last enzyme in the tocopherol biosynthetic pathway and it catalyzes the conversion of γ-tocopherol into α-tocopherol, the nutritionally significant and most bioactive form of vitamin E. Although the γ-TMT gene has been successfully overexpressed in many crops to enhance their α-tocopherol content but still only few attempts have been made to uncover its structural, functional and regulation aspects at protein level. In this study, we have cloned the complete 909bp coding sequence of Glycine max γ-TMT (Gm γ-TMT) gene that encodes the corresponding protein comprising of 302 amino acid residues...
August 12, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28811215/molecular-mechanisms-underlying-protection-against-h9n2-influenza-virus-challenge-in-mice-by-recombinant-lactobacillus-plantarum-with-surface-displayed-ha2-ltb
#2
Yanlong Jiang, Guilian Yang, Qi Wang, Zhannan Wang, Wentao Yang, Wei Gu, Chunwei Shi, Jianzhong Wang, Haibin Huang, Chunfeng Wang
It has been considered that the Avian influenza virus (AIV) causes severe threats to poultry industry. In this study, we constructed a series of recombinant Lactobacillus plantarum (L. plantarum) with surface displayed hemagglutinin subunit 2 (HA2) alone or together with heat-labile toxin B subunit (LTB) from enterotoxigenic Escherichia coli. Balb/c mice were used as model to evaluate the protective effects of recombinant L. plantarum strains against H9N2 subtype challenge. The results showed that the presence of LTB significantly increased the percentages of CD3(+)CD4(+)IL-4(+), CD3(+)CD4(+)IFN-γ(+) and CD3(+)CD4(+)IL-17(+) T cells, as well as CD3(+)CD8(+)IFN-γ(+) T cells in spleen and MLNs determined by Fluorescence-Activated Cell Sorting assay...
August 12, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28811153/the-design-of-target-specific-antibodies-scfv-by-applying-de-novo-workflow-case-study-on-bmr1-antigen-from-brugia-malayi
#3
Bee Yin Khor, Theam Soon Lim, Rahmah Noordin, Yee Siew Choong
De novo approach was applied to design single chain fragment variable (scFv) for BmR1, a recombinant antigen from Bm17DIII gene which is the primary antigen used for the detection of anti-BmR1 IgG4 antibodies in the diagnostic of lymphatic filariasis. Three epitopes of the BmR1 was previously predicted form an ab initio derived three-dimensional structure. A collection of energetically favourable conformations was generated via hot-spot-centric approach. This resulted in a set of three different scFv scaffolds used to compute the high shape complementary conformations via dock-and-design approach with the predicted epitopes of BmR1...
July 19, 2017: Journal of Molecular Graphics & Modelling
https://www.readbyqxmd.com/read/28810661/identification-of-lectin-counter-receptors-on-cell-membranes-by-proximity-labeling
#4
Gang Wu, Manjula Nagala, Paul R Crocker
Lectin-glycan interactions play important roles in many biological systems, but the nature of glycoprotein counter-receptors expressed on cell membranes is often poorly understood. To help overcome this problem, we developed a method based on proximity labeling technology. Using a peroxidase-coupled lectin, addition of H2O2 and tyramide-biotin substrates leads to generation of short-range biotin radicals that biotinylate proteins in the immediate vicinity of the bound lectin, which can subsequently be identified...
September 1, 2017: Glycobiology
https://www.readbyqxmd.com/read/28809993/adalimumab-in-chronic-plaque-psoriasis-a-clinical-guide
#5
Jashin J Wu, W C Valdecantos
<p>Psoriasis is a common, inflammatory disease that manifests itself as lesions on the skin, which greatly impacts the physical and psychological wellbeing of those affected. The current goal of treatment in psoriasis is to improve the signs and symptoms of disease, whilst minimizing the burden of disease on patient health-related quality of life. Psoriasis can also be associated with other comorbidities such as joint disease, cardiovascular disease, and depression, which can add to the complexity of treatment...
August 1, 2017: Journal of Drugs in Dermatology: JDD
https://www.readbyqxmd.com/read/28809150/a-tupaia-paramyxovirus-vector-system-for-targeting-and-transgene-expression
#6
Christine E Engeland, Sascha Bossow, Andrew W Hudacek, Birgit Hoyler, Judith Förster, Rūta Veinalde, Dirk Jäger, Roberto Cattaneo, Guy Ungerechts, Christoph Springfeld
Viruses from the diverse family of Paramyxoviridae include important pathogens and are applied in gene therapy and for cancer treatment. The Tupaia paramyxovirus (TPMV), isolated from the kidney of a tree shrew, does not infect human cells and neutralizing antibodies against other Paramyxoviridae do not cross-react with TPMV. Here, we present a vector system for de novo generation of infectious TPMV that allows for insertion of additional genes as well as targeting using antibody single-chain variable fragments...
August 15, 2017: Journal of General Virology
https://www.readbyqxmd.com/read/28808918/idarucizumab-in-dabigatran-treated-patients-with-acute-ischemic-stroke-receiving-alteplase-a-systematic-review-of-the-available-evidence
#7
REVIEW
Slaven Pikija, Laszlo K Sztriha, J Sebastian Mutzenbach, Stefan M Golaszewski, Johann Sellner
BACKGROUND AND PURPOSE: Current guidelines do not recommend the use of intravenous recombinant tissue plasminogen activator in patients with acute ischemic stroke who receive direct oral anticoagulants. While the humanized monoclonal antibody idarucizumab can quickly reverse the anticoagulant effects of the thrombin inhibitor dabigatran, safety data for subsequent tissue plasminogen activator treatment are sparse. Here, we review current knowledge about dabigatran reversal prior to systemic reperfusion treatment in acute ischemic stroke...
August 14, 2017: CNS Drugs
https://www.readbyqxmd.com/read/28808292/binding-of-the-helicobacter-pylori-oipa-causes-apoptosis-of-host-cells-via-modulation-of-bax-bcl-2-levels
#8
Omid Teymournejad, Ashraf Mohabati Mobarez, Zuhair Mohammad Hassan, Amin Talebi Bezmin Abadi
The H. pylori outer inflammatory protein A (OipA) is an outer membrane protein that contributes to gastric inflammation. OipA is believed to affect intra-cellular signalling and modulate the host signalling pathways. The aim of the current study was to clarify the role of OipA in H. pylori pathogenesis and its effect on host cell signalling pathways. To this end, the oipA gene was isolated and inserted into cloning and expression vectors. The recombinant plasmid was transferred into an expression host to produce OipA, which was subsequently purified by affinity chromatography and used for antibody production...
August 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28807316/vaccination-with-eimeria-tenella-elongation-factor-1%C3%AE-recombinant-protein-induces-protective-immunity-against-e-tenella-and-e-maxima-infections
#9
Rui-Qing Lin, Hyun S Lillehoj, Seung Kyoo Lee, Sungtaek Oh, Alfredo Panebra, Erik P Lillehoj
Avian coccidiosis is caused by multiple species of the apicomplexan protozoan, Eimeria, and is one of the most economically devastating enteric diseases for the poultry industry worldwide. Host immunity to Eimeria infection, however, is relatively species-specific. The ability to immunize chickens against different species of Eimeria using a single vaccine will have a major beneficial impact on commercial poultry production. In this paper, we describe the molecular cloning, purification, and vaccination efficacy of a novel Eimeria vaccine candidate, elongation factor-1α (EF-1α)...
August 30, 2017: Veterinary Parasitology
https://www.readbyqxmd.com/read/28807275/recombinant-miro-domain-containing-protein-of-haemonchus-contortus-rmiro-1-activates-goat-peripheral-blood-mononuclear-cells-in-vitro
#10
YuLing Wen, YuJian Wang, WenJuan Wang, MingMin Lu, Muhammad Ehsan, XiaoWei Tian, RuoFeng Yan, XiaoKai Song, LiXin Xu, XiangRui Li
In our previous proteomics study, we identified Miro domain-containing protein (Miro-1), an excretory and secretory product of the pole worm, Haemonchus contortus, binds to goat peripheral blood mononuclear cells (PBMCs) in vivo. However, our understanding of the role of Miro-1and its potential immune impact on goat PBMCs is limited. The aim of the present study was to evaluate the effects of Miro-1 on functions of goat PBMCs in vitro. Recombinant protein (rMiro-1) was expressed in a prokaryote and incubated with goat PBMCs...
August 30, 2017: Veterinary Parasitology
https://www.readbyqxmd.com/read/28807043/an-equine-herpesvirus-type-1-ehv-1-vector-expressing-rift-valley-fever-virus-rvfv-gn-and-gc-induces-neutralizing-antibodies-in-sheep
#11
Abdelrahman Said, Mona Elmanzalawy, Guanggang Ma, Armando Mario Damiani, Nikolaus Osterrieder
Rift Valley fever virus (RVFV) is an arthropod-borne bunyavirus that can cause serious and fatal disease in humans and animals. RVFV is a negative-sense RNA virus of the Phlebovirus genus in the Bunyaviridae family. The main envelope RVFV glycoproteins, Gn and Gc, are encoded on the M segment of RVFV and known inducers of protective immunity. In an attempt to develop a safe and efficacious RVF vaccine, we constructed and tested a vectored equine herpesvirus type 1 (EHV-1) vaccine that expresses RVFV Gn and Gc...
August 14, 2017: Virology Journal
https://www.readbyqxmd.com/read/28806153/expression-purification-and-refolding-of-human-lipocalin-6-and-production-of-a-monoclonal-antibody-against-this-protein
#12
Jiong Chen, Wei Feng, Yue Zhao, Yaqin Li, Fei Zhan
Human lipocalin 6 (hLCN6) is a member of the lipocalin family, which is a group of structurally conserved hydrophobic ligand binding proteins, and widely distributed in animal, plant, and bacteria. Specific expression of hLCN6 in the epididymis and localization of this protein on the surface of spermatozoa suggest a role played by hLCN6, which may function as a transporter to carry ligands in the epididymal channel. However, the role of hLCN6 in sperm maturation has been largely unknown due to the lack of effective antibodies...
August 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28806151/establishment-of-a-monoclonal-antibody-against-chgilz
#13
Zhiyuan He, Yongqiang Wang
The chicken glucocorticoid-induced leucine zipper (chGILZ) participates in the inflammation of avian immunosuppressive diseases. We aimed to establish a monoclonal antibody (MAb) against chGILZ and to investigate its distribution in clinical diagnosis. A gene cloned from chicken embryo fibroblast (CEF) cell was inserted into the expression vector pET-28a and pGEX-6p-1. The recombinant expression vectors were transformed into Escherichia coli BL21 (DE3). Then the recombinant proteins His-chGILZ and GST-chGILZ were successfully expressed and purified...
August 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28805498/gingiskhan%C3%A2-protease-cleavage-allows-a-high-throughput-antibody-to-fab-conversion-enabling-direct-functional-assessment-during-lead-identification-of-human-monoclonal-and-bispecific-igg1-antibodies
#14
Jörg Moelleken, Manuel Endesfelder, Christian Gassner, Sabine Lingke, Simone Tomaschek, Oksana Tyshchuk, Stefan Lorenz, Ulrike Reiff, Michael Mølhøj
The determination of the binding strength of immunoglobulins (IgGs) to targets can be influenced by avidity when the targets are soluble di- or multimeric proteins, or associated to cell surfaces, including surfaces introduced from heterogeneous assays. However, for the understanding of the contribution of a second drug-to-target binding site in molecular design, or for ranking of monovalent binders during lead identification, affinity-based assessment of the binding strength is required. Typically, monovalent binders like antigen-binding fragments (Fabs) are generated by proteolytic cleavage with papain, which often results in a combination of under- and over-digestion, and requires specific optimization and chromatographic purification of the desired Fabs...
August 14, 2017: MAbs
https://www.readbyqxmd.com/read/28805287/nongenetically-modified-lactococcus-lactis-adjuvanted-vaccination-enhanced-innate-immunity-against-helicobacter-pylori
#15
Wei Liu, Zhoulin Tan, Hai Liu, Zhiqin Zeng, Shuanghui Luo, Huimin Yang, Lufeng Zheng, Tao Xi, Yingying Xing
BACKGROUND: Gram-positive enhancer matrix particles (GEM) produced by Lactococcus lactis can enhance vaccine-induced immune response. However, the mechanism under which this adjuvant mounts the efficacy of orally administered vaccines remains unexplored. MATERIALS AND METHODS: We used a prophylactic mice model to investigate the mechanism of GEM-adjuvanted vaccination. Helicobacter pylori urease-specific antibody response was monitored and detected in murine serum by ELISA...
August 14, 2017: Helicobacter
https://www.readbyqxmd.com/read/28804955/3m-052-as-an-adjuvant-for-a-plga-microparticle-based-leishmania-donovani-recombinant-protein-vaccine
#16
Qian Wang, Meagan A Barry, Christopher A Seid, Elissa M Hudspeth, C Patrick McAtee, Michael J Heffernan
It is believed that an effective vaccine against leishmaniasis will require a T helper type 1 (TH 1) immune response. In this study, we investigated the adjuvanticity of the Toll-like receptor (TLR) 7/8 agonist 3M-052 in combination with the Leishmania donovani 36-kDa nucleoside hydrolase recombinant protein antigen (NH36). NH36 and 3M-052 were encapsulated in separate batches of poly(lactic-co-glycolic acid) (PLGA) microparticles (MPs). The loading efficiency for NH36 was 83% and for 3M-052 was above 95%. In vitro stimulation of bone marrow-derived dendritic cells, measured by IL-12 secretion, demonstrated that 3M-052 (free or MP-formulated) had a concentration-dependent immunostimulatory effect with an optimum concentration of 2 µg/mL...
August 14, 2017: Journal of Biomedical Materials Research. Part B, Applied Biomaterials
https://www.readbyqxmd.com/read/28804866/virally-vectored-vaccine-delivery-medical-needs-mechanisms-advantages-and-challenges
#17
Daniel D Pinschewer
Vaccines represent one of the most successful chapters in the history of medicine. Over the past decades, the advent of recombinant cDNA technology has enabled the biomedical community to genetically engineer viruses for vaccine delivery purposes. As a starting point, this review evaluates the unmet medical needs, which drive scientists and industry to exploit such fundamentally new technology for human vaccination. The author discusses the molecular functioning, production and safety profile of replication-competent and -deficient viral vector systems, representing two fundamentally distinct classes of "genetic vaccines"...
August 14, 2017: Swiss Medical Weekly
https://www.readbyqxmd.com/read/28803862/recombinant-lcmv-vectors-induce-protective-immunity-following-homologous-and-heterologous-vaccinations
#18
Jessica Wingerath, Dmitrij Ostroumov, Norman Woller, Michael P Manns, Daniel D Pinschewer, Klaus Orlinger, Ursula Berka, Florian Kühnel, Thomas C Wirth
Successful vaccination against cancer and infectious diseases relies on the induction of adaptive immune responses that induce high-titer antibodies or potent cytoxic T cell responses. In contrast to humoral vaccines, the amplification of cellular immune responses is often hampered by anti-vector immunity that either pre-exists or develops after repeated homologous vaccination. Replication-defective lymphocytic choriomeningitis virus (LCMV) vectors represent a novel generation of vaccination vectors that induce potent immune responses while escaping recognition by neutralizing antibodies...
July 20, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28803502/immunogenicity-antibody-persistence-and-safety-of-the-60-%C3%AE-g-hepatitis-b-vaccine-in-hemodialysis-patients-a-multicenter-randomized-double-blind-parallel-controlled-trial
#19
Yongliang Feng, Xiaohong Shi, Jing Shi, Linying Gao, Guangming Liu, Yanpeng Cheng, Minghu Pan, Chunxia Li, Jun Wang, Xuxia Guo, Yawei Zhang, Xiaofeng Liang, Suping Wang
OBJECTIVES: To evaluate the immunogenicity, antibody persistence, and safety of the 60µg hepatitis B vaccine in hemodialysis patients in China. METHODS: We conducted a multicenter, randomized, double-blind, parallel-controlled trial including 352 hemodialysis patients who were centrally randomized in a ratio of 1:1 to receive a 20 µg (IM20 group) or 60 µg (IM60 group) recombinant hepatitis B vaccine at months 0, 1, and 6. RESULTS: The vaccine-elicited antibody responses peaked at month 7, and declined at month 12...
August 14, 2017: Expert Review of Vaccines
https://www.readbyqxmd.com/read/28802994/expression-and-purification-of-norovirus-virus-like-particles-in-escherichia-coli-and-their-immunogenicity-in-mice
#20
Yuqi Huo, Xin Wan, Tong Ling, Jie Wu, Wenhui Wang, Shuo Shen
Norovirus (NoV) virus like particles (VLPs) produced in Spodoptera frugiperda (Sf9) cells have been tested in human volunteers as vaccine candidate and were shown to be protective against NoV induced acute gastroenteritis. In this study, prevailing Sydney-2012-like NoV major capsid protein gene with or without N-terminal deletions (N26 and N38, 26 and 38 amino acids deleted from N terminus,respectively) were sub-cloned into prokaryotic expression vector, pCold III and pCold IV. Soluble and insoluble proteins were detected for both vectors after induction and higher levels of protein expression were observed for constructs pCold III-N26 and pCold III-N38...
August 9, 2017: Molecular Immunology
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