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https://www.readbyqxmd.com/read/28225865/continuous-blood-purification-treatment-for-endotoxin-induced-acute-respiratory-distress-syndrome
#1
Y Jiang, R Lin, Y Xu, S Zhang, K Cui, M Zhu, A Li, C Chen, J Yang, W Yang
This study aimed to explore the effects of continuous blood purification (CBP) treatment in pigs affected with acute respiratory distress syndrome (ARDS). A total of 12 healthy male pigs, weighing 12±1.8 kg, were randomly and equally assigned to the control and experimental groups. The ARDS pig model was prepared by intravenous injections of endotoxin (20 µg/kg). The control group was given conventional supportive therapy, while the experimental group was given continuous veno-venous hemofiltration therapy...
February 16, 2017: Brazilian Journal of Medical and Biological Research, Revista Brasileira de Pesquisas Médicas e Biológicas
https://www.readbyqxmd.com/read/28225803/bacterial-expression-correct-membrane-targeting-and-functional-folding-of-the-hiv-1-membrane-protein-vpu-using-a-periplasmic-signal-peptide
#2
Arpan Deb, William A Johnson, Alexander P Kline, Boston J Scott, Lydia R Meador, Dustin Srinivas, Jose M Martin-Garcia, Katerina Dörner, Chad R Borges, Rajeev Misra, Brenda G Hogue, Petra Fromme, Tsafrir S Mor
Viral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors. But key structure/function questions remain regarding the mechanisms by which the Vpu protein contributes to HIV-1 pathogenesis. Here we describe expression of Vpu in bacteria, its purification and characterization. We report the successful expression of PelB-Vpu in Escherichia coli using the leader peptide pectate lyase B (PelB) from Erwinia carotovora...
2017: PloS One
https://www.readbyqxmd.com/read/28225217/proteomic-analysis-of-the-cullin4b-interactome-using-proximity-dependent-biotinylation-in-living-cells
#3
Hailong Zhang, Shupeng Li, Pingting Liu, Frankie H F Lee, Albert H C Wong, Fang Liu
Cullin 4B (CUL4B) mutations have been implicated in mental retardation and dopamine-related behaviors due to disruptions in their interaction with Cullin-RING E3 ligases (CRLs). Thus, further identification of CUL4B substrates can increase the knowledge of protein homeostasis and illuminate the role of CUL4B in neuropsychiatric disease. However, the transient nature of the coupling between CUL4B and its substrates is difficult to detect in vivo using current approaches, thus hampers efforts to investigate functions of CRLs within unperturbed living systems...
February 22, 2017: Proteomics
https://www.readbyqxmd.com/read/28225144/molecular-and-genetic-basis-for-early-stage-structural-diversifications-in-hapalindole-type-alkaloid-biogenesis
#4
Qin Zhu, Xinyu Liu
Heterologous expressions and purifications of all WelU proteins from the welwitindolinone pathways in Hapalosiphon welwitschii UTEX B1830 and IC-52-3 led to the discovery that WelU1 and WelU3 selectively assemble 12-epi-fischerindole U (2) and 12-epi-hapalindole C (1), respectively, from 3-geranyl 3-isocyanovinyl indolenine (4) via an enzymatic cascade featuring the Cope rearrangement, stereoselective aza-Prins cyclization and regioselective carbocation deposition. In combination with the in vitro characterization of WelU1/WelU3-homolog AmbU4 for the biogenesis of 12-epi-hapalindole U, this study provide a unified view on the origin of the early stage structural diversifications in hapalindole-type alkaloid biosynthesis, post common intermediate 4...
February 22, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28225020/improved-antitumor-activity-of-trail-fusion-protein-via-formation-of-self-assembling-nanoparticle
#5
Kaizong Huang, Ningjun Duan, Chunmei Zhang, Ran Mo, Zichun Hua
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been known as a promising agent for cancer therapy due to its specific apoptosis-inducing effect on tumor cells rather than most normal cells. However, systemically delivered TRAIL suffers from a rapid clearance from the body with an extremely short half-life. Thermally responsive elastin-like polypeptides (ELPs) are a promising class of temperature sensitive biopolymers based on the structural motif found in mammalian tropoelastin and retain the advantages of polymeric drug delivery systems...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28224978/methylated-antibody-affinity-purification-to-improve-proteomic-identification-of-plant-rna-polymerase-pol-v-complex-and-the-interacting-proteins
#6
Guochen Qin, Jun Ma, Xiaomei Chen, Zhaoqing Chu, Yi-Min She
Affinity purification followed by enzymatic digestion and mass spectrometry has been widely utilized for the sensitive detection of interacting proteins and protein complexes in various organisms. In plants, the method is technically challenging due to the low abundance proteins, non-specific binding and difficulties of eluting interacting proteins from antibody beads. In this report, we describe a strategy to modify antibodies by reductive methylation of lysines without affecting their binding properties, followed by on-bead digestion of bound proteins with endoproteinase Lys-C...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28224267/selection-of-dna-aptamers-for-extra-cellular-domain-of-human-epidermal-growth-factor-receptor-2-to-detect-her2-positive-carcinomas
#7
A Sett, B B Borthakur, U Bora
BACKGROUND: Human epidermal growth factor receptor 2 (Her2, an orphan receptor of ErbB family) is considered as an important biomarker as it plays a key role in the development and progression of aggressive types of breast, ovarian, stomach and gastric cancer. In the present study, we developed novel DNA aptamers against the extra-cellular domain (ECD) of Her2 protein for detection of Her2-positive carcinomas. METHODS: We cloned and expressed Her2-ECD protein in E...
February 21, 2017: Clinical & Translational Oncology
https://www.readbyqxmd.com/read/28223717/scale-up-and-pharmacokinetic-study-of-a-novel-mutated-chimeric-tissue-plasminogen-activator-mt-pa-in-rats
#8
Mozhgan Raigani, Mohammad-Reza Rouini, Ali-Akbar Golabchifar, Esmat Mirabzadeh, Behrouz Vaziri, Farzaneh Barkhordari, Fatemeh Davami, Fereidoun Mahboudi
Because of high mortality caused by cardiovascular diseases, various fibrinolytic agents with diverse pharmacokinetic and pharmacodynamic properties have been developed. A novel mutated chimeric tissue plasminogen activator (mt-PA) was developed by the removal of first three domains of t-PA, insertion of GHRP sequence and mutation towards resistance to plasminogen activator inhibitor-1 (PAI-1). Mt-PA protein was expressed in Expi293F cells. The expression level of mt-PA was found to be 5000 IU/mL. Following purification, the pharmacokinetic properties of mt-PA were evaluated in three doses in rats...
February 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28223313/active-multi-enzyme-assemblies-for-long-chain-olefinic-hydrocarbon-biosynthesis
#9
James K Christenson, Matthew R Jensen, Brandon R Goblirsch, Fatuma Mohamed, Wei Zhang, Carrie M Wilmot, Lawrence P Wackett
Bacteria from different Phyla produce long-chain olefinic hydrocarbons derived from an OleA-catalyzed Claisen condensation of two fatty acyl-CoA substrates, followed by reduction and oxygen elimination reactions catalyzed by the proteins OleB, OleC, and OleD. In this report, OleA, OleB, OleC, and OleD were individually purified as soluble proteins, and all were found to be essential for reconstituting hydrocarbon biosynthesis. Recombinant co-expression of tagged OleABCD proteins from Xanthomonas campestris in Escherichia coli and purification over His6x- and FLAG-columns resulted in OleA separating whilst OleBCD purified together, irrespective of which of the four Ole proteins were tagged...
February 21, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28222997/integrated-analysis-of-recombinant-bpv-1-l1-protein-for-the-production-of-a-bovine-papillomavirus-vlp-vaccine
#10
Diego Grando Módolo, Rodrigo Pinheiro Araldi, Jacqueline Mazzuchelli-de-Souza, Alexandre Pereira, Daniel Carvalho Pimenta, Letícia Maria Zanphorlin, Willy Beçak, Marcelo Menossi, Rita de Cassia Stocco, Rodrigo Franco de Carvalho
Bovine papillomatosis is an infectious disease that is caused by bovine papillomavirus (BPV), which results in important economic losses. However, no BPV vaccines or effective treatment methods are commercially available to date. Moreover, the absence of papillomavirus replication in vitro makes the use of recombinant protein a promising candidate for vaccine formulations. Hence, we developed an integrated study on the L1 capsid protein of BPV-1, obtained from a bacterial expression system, regarding its purification, biosafety, thermostability and immunogenicity...
February 17, 2017: Vaccine
https://www.readbyqxmd.com/read/28222503/selenium-levels-in-serum-red-blood-cells-and-cerebrospinal-fluid-of-alzheimer-s-disease-patients-a-report-from-the-australian-imaging-biomarker-lifestyle-flagship-study-of-ageing-aibl
#11
Bárbara R Cardoso, Dominic J Hare, Ashley I Bush, Qiao-Xin Li, Christopher J Fowler, Colin L Masters, Ralph N Martins, Katherine Ganio, Amber Lothian, Soumya Mukherjee, Eugene A Kapp, Blaine R Roberts
Selenium (Se) protects cells against oxidative stress damage through a range of bioactive selenoproteins. Increased oxidative stress is a prominent feature of Alzheimer's disease (AD), and previous studies have shown that Se deficiency is associated with age-related cognitive decline. In this study, we assessed Se status in different biofluids from a subgroup of participants in the Australian Imaging, Biomarkers and Lifestyle Flagship Study of Ageing. As Se in humans can either be an active component of selenoproteins or inactive via non-specific incorporation into other proteins, we used both size exclusion chromatography-inductively coupled plasma-mass spectrometry (SEC-ICP-MS) and tandem mass spectrometry to characterize selenoproteins in serum...
February 10, 2017: Journal of Alzheimer's Disease: JAD
https://www.readbyqxmd.com/read/28222187/a-purified-maa-based-elisa-is-a-useful-tool-for-determining-anti-maa-antibody-titer-with-high-sensitivity
#12
Takasumi Shimomoto, Leonard B Collins, Xianwen Yi, Darcy W Holley, Zhenfa Zhang, Xu Tian, Koji Uchida, Chunguang Wang, Sohvi Hörkkö, Monte S Willis, Avram Gold, Scott J Bultman, Jun Nakamura
Atherosclerosis is widely accepted to be a chronic inflammatory disease, and the immunological response to the accumulation of LDL is believed to play a critical role in the development of this disease. 1,4-Dihydropyridine-type MAA-adducted LDL has been implicated in atherosclerosis. Here, we have demonstrated that pure MAA-modified residues can be chemically conjugated to large proteins without by-product contamination. Using this pure antigen, we established a purified MAA-ELISA, with which a marked increase in anti-MAA antibody titer was determined at a very early stage of atherosclerosis in 3-month ApoE-/- mice fed with a normal diet...
2017: PloS One
https://www.readbyqxmd.com/read/28220817/a-simple-approach-for-preparation-of-affinity-matrices-simultaneous-purification-and-reversible-immobilization-of-a-streptavidin-mutein-to-agarose-matrix
#13
Sau-Ching Wu, Chris Wang, Dave Hansen, Sui-Lam Wong
SAVSBPM18 is an engineered streptavidin for affinity purification of both biotinylated biomolecules and recombinant proteins tagged with streptavidin binding peptide (SBP) tags. To develop a user-friendly approach for the preparation of the SAVSBPM18-based affinity matrices, a designer fusion protein containing SAVSBPM18 and a galactose binding domain was engineered. The galactose binding domain derived from the earthworm lectin EW29 was genetically modified to eliminate a proteolytic cleavage site located at the beginning of the domain...
February 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28220811/aptamers-for-respiratory-syncytial-virus-detection
#14
Krisztina Percze, Zoltán Szakács, Éva Scholz, Judit András, Zsuzsanna Szeitner, Corné H van den Kieboom, Gerben Ferwerda, Marien I de Jonge, Róbert E Gyurcsányi, Tamás Mészáros
The identification of the infectious agents is pivotal for appropriate care of patients with viral diseases. Current viral diagnostics rely on selective detection of viral nucleic acid or protein components. In general, detection of proteins rather than nucleic acids is technically more suitable for rapid tests. However, protein-based virus identification methods depend on antibodies limiting the practical applicability of these approaches. Aptamers rival antibodies in target selectivity and binding affinity, and excel in terms of robustness and cost of synthesis...
February 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28220544/prospective-purification-and-characterization-of-m%C3%A3-ller-glia-in-the-mouse-retina-regeneration-assay
#15
Patrick Schäfer, Mike O Karl
Reactive gliosis is an umbrella term for various glia functions in neurodegenerative diseases and upon injury. Specifically, Müller glia (MG) in some species readily regenerate retinal neurons to restore vision loss after insult, whereas mammalian MG respond by reactive gliosis-a heterogeneous response which frequently includes cell hypertrophy and proliferation. Limited regeneration has been stimulated in mammals, with a higher propensity in young MG, and in vitro compared to in vivo, but the underlying processes are unknown...
February 21, 2017: Glia
https://www.readbyqxmd.com/read/28220411/peptidoglycan-isolation-and-binding-studies-with-lysm-type-pattern-recognition-receptors
#16
Ute Bertsche, Andrea A Gust
In the last decade, more and more plant receptors for complex carbohydrate structures have been described. However, studies on receptor binding to glycan ligands are often hampered due to the technical challenge to obtain pure preparations of homogeneous carbohydrate ligands such as bacterial peptidoglycan (PGN) in amounts suitable for studying protein-glycan interactions. Also, most approaches rely on the availability of defined soluble ligands, which in the case of glycans can rarely be synthesized but have to be purified from the respective microorganism...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28220352/purification-characterization-and-overexpression-of-an-endo-1-4-%C3%AE-mannanase-from-thermotolerant-bacillus-sp-swu60
#17
Weeranuch Seesom, Polphet Thongket, Tomohiro Yamamoto, Shigeo Takenaka, Tatsuji Sakamoto, Wasana Sukhumsirichart
Endo-β-1,4-mannanases are important catalytic agents in several industries. The enzymes randomly cleave the β-1,4-linkage in the mannan backbone and release short β-1,4-mannooligosaccharides and mannose. In the present study, mannanase (ManS2) from thermotolerant Bacillus sp. SWU60 was purified, characterized, and its gene was cloned and overexpressed in Escherichia coli. ManS2 was purified from culture filtrate (300 ml) by using hydrophobic, ion-exchange, and size-exclusive liquid chromatography. The apparent molecular mass was 38 kDa...
March 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28220242/expression-of-clonorchis-sinensis-giiispla2-protein-in-baculovirus-infected-insect-cells-and-its-overexpression-facilitating-epithelial-mesenchymal-transition-in-huh7-cells-via-akt-pathway
#18
Mei Shang, Zhizhi Xie, Zeli Tang, Lei He, Xiaoyun Wang, Caiqin Wang, Yinjuan Wu, Ye Li, Lu Zhao, Zhiyue Lv, Zhongdao Wu, Yan Huang, Xinbing Yu, Xuerong Li
Although prior studies confirmed that group III secretory phospholipase A2 of Clonorchis sinensis (CsGIIIsPLA2) had stimulating effect on liver fibrosis by binding to LX-2 cells, large-scale expression of recombinant protein and its function in the progression of hepatoma are worth exploring. Because of high productivity and low lipopolysaccharides (LPS) in the Sf9-baculovirus expression system, we firstly used this system to express the coding region of CsGIIIsPLA2. The molecular weight of recombinant CsGIIIsPLA2 protein was about 34 kDa...
February 20, 2017: Parasitology Research
https://www.readbyqxmd.com/read/28218978/biochemical-characterization-of-metabolism-based-atrazine-resistance-in-amaranthus-tuberculatus-and-identification-of-an-expressed-gst-associated-with-resistance
#19
Anton F Evans, Sarah R O'Brien, Rong Ma, Aaron G Hager, Chance W Riggins, Kris N Lambert, Dean E Riechers
Rapid detoxification of atrazine in naturally tolerant crops such as maize (Zea mays) and grain sorghum (Sorghum bicolor) results from glutathione S-transferase (GST) activity. In previous research, two atrazine-resistant waterhemp (Amaranthus tuberculatus) populations from Illinois, U.S.A. (designated ACR and MCR) displayed rapid formation of atrazine-glutathione (GSH) conjugates, implicating elevated rates of metabolism as the resistance mechanism. Our main objective was to utilize protein purification combined with qualitative proteomics to investigate the hypothesis that enhanced atrazine detoxification, catalyzed by distinct GSTs, confers resistance in ACR and MCR...
February 20, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28218648/expression-purification-and-monitoring-of-conformational-changes-of-hcb2-tmh67h8-in-different-membrane-mimetic-lipid-mixtures-using-circular-dichroism-and-nmr-techniques
#20
Elvis K Tiburu, Jianqin Zhuang, Heidimarie N A Fleischer, Patrick K Arthur, Gordon A Awandare
This work was intended to develop self-assembly lipids for incorporating G-protein coupled receptors (GPCRs) in order to improve the success rate for nuclear magnetic resonance spectroscopy (NMR) structural elucidation. We hereby report the expression and purification of uniformly (15)N-labeled human cannabinoid receptor-2 domain in insect cell media. The domain was refolded by screening several membrane mimetic environments. Different q ratios of isotropic bicelles were screened for solubilizing transmembrane helix 6, 7 and 8 (TMH67H8)...
February 17, 2017: Membranes
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