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https://www.readbyqxmd.com/read/27925690/purification-and-characterization-of-a-platelet-aggregation-inhibitor-and-anticoagulant-cc-5_ntase-cd-73-like-from-cerastes-cerastes-venom
#1
Samah Saoud, Fatah Chérifi, Traki Benhassine, Fatima Laraba-Djebari
The present study is the first attempt to report the characterization of a nucleotidase from Cerastes cerastes venom. A 70 kDa 5'-nucleotidase (Cc-5'NTase) was purified to homogeneity. The amino acid sequence of Cc-5'NTase displayed high homology with many nucleotidases. Its activity was optimal at pH 7 with a specific hydrolytic activity toward mono-, di-, and triphosphate adenylated nucleotides. Cc-5'NTase preferentially hydrolyzed ADP and obeyed Michaelis-Menten kinetics. Among the metals and inhibitors tested, Ni(2+) and Mg(2+) completely potentiated enzyme activity, whereas EGTA, PMSF, iodoacetamide, vanillic acid, vanillyl mandelic acid, and 1,10-phenanthroline partially abolished its activity...
December 7, 2016: Journal of Biochemical and Molecular Toxicology
https://www.readbyqxmd.com/read/27925666/enriching-s-4-u-rna-using-methane-thiosulfonate-mts-chemistry
#2
Erin E Duffy, Matthew D Simon
Metabolic labeling of cellular RNA is a useful approach to study RNA biology. 4-Thiouridine (s(4) U) is a convenient nucleoside for metabolic labeling because it is cell permeable and is incorporated into newly transcribed RNA, and the sulfur moiety provides a handle for biochemical purification. However, a critical step in the purification of s(4) U-RNA is the efficiency of the chemistry used to enrich s(4) U-RNA. Here, we present a protocol for s(4) U-RNA enrichment that includes efficient and reversible covalent chemistry to biotinylate s(4) U-RNA using the activated disulfide methane thiosulfonate conjugated to biotin (MTS-biotin), followed by enrichment on streptavidin beads...
December 7, 2016: Current Protocols in Chemical Biology
https://www.readbyqxmd.com/read/27925294/improved-68-ga-labeling-method-using-ethanol-addition-application-to-the-%C3%AE-helical-peptide-dota-famp
#3
Koki Hasegawa, Emi Kawachi, Yoshinari Uehara, Tsuyoshi Yoshida, Satoshi Imaizumi, Masahiro Ogawa, Shin-Ichiro Miura, Keijiro Saku
We examined the (68) Ga labeling of the α-helical peptide, DOTA-FAMP, and evaluated conformational changes during radiolabeling. (68) Ga-DOTA-FAMP is a positron emission tomography probe candidate for atherosclerotic plaques. The labeling yield achieved by Zhernosekov's method (using acetone for (68) Ga purification) was compared with that achieved by the original and 2 modified Mueller's methods (using NaCl solution). Modified method I involves desalting the (68) Ga prior to labeling, and modified method II involves the inclusion of ethanol in the labeling solution...
December 7, 2016: Journal of Labelled Compounds & Radiopharmaceuticals
https://www.readbyqxmd.com/read/27924601/a-protocol-for-the-determination-of-the-methylation-status-of-gingival-tissue-dna-at-specific-cpg-islands
#4
Trudy J Milne
Tissue biopsies are very precious. A method that allows the isolation of a high quality and quantity of genomic DNA, total RNA, and total protein from a single biopsy that is suitable for downstream applications (e.g., DNA methylation analysis, quantitative PCR, and gel electrophoresis techniques) is very desirable. The method described here combines a tissue stabilization reagent combined with a spin-column method for the simultaneous purification of gingival tissue DNA, RNA, and protein. The genomic DNA is then used for quantitative analysis of DNA methylation using real-time PCR (the qAMP method)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924596/methods-to-study-antagonistic-activities-among-oral-bacteria
#5
Fengxia Qi, Jens Kreth
Most bacteria in nature exist in multispecies communities known as biofilms. In the natural habitat where resources (nutrient, space, etc.) are usually limited, individual species must compete or collaborate with other neighboring species in order to perpetuate in the multispecies community. The human oral cavity is colonized by >700 microbial species known as the indigenous microbiota. This indigenous flora normally maintains an ecological balance through antagonistic as well as mutualistic interspecies interactions...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924587/qualitative-and-quantitative-proteome-analysis-of-oral-fluids-in-health-and-periodontal-disease-by-mass-spectrometry
#6
Erdjan Salih
The significance of protein identification and characterization by classical protein chemistry approaches is clearly highlighted by our detailed understanding of the biological systems assembled over a time period of almost a century. The advent of state-of-the-art mass spectrometry (MS) with sensitivity, speed, and global protein analysis capacity without individual protein purification has transformed the classical protein chemistry with premise to accelerate discovery. These combined with the ability of the oral fluids such as whole saliva (WS) and gingival crevicular fluid (GCF) to reflect both systemic and locally derived proteins have generated significant interest to characterize these fluids more extensively by MS technology...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924562/single-step-affinity-purification-of-erk-signaling-complexes-using-the-streptavidin-binding-peptide-sbp-tag
#7
Liu Yang, Alexey Veraksa
Elucidation of biological functions of signaling proteins is facilitated by studying their protein-protein interaction networks. Affinity purification combined with mass spectrometry (AP-MS) has become a favorite method to study protein complexes. Here we describe a procedure for single-step purification of ERK (Rolled) and associated proteins from Drosophila cultured cells. The use of the streptavidin-binding peptide (SBP) tag allows for a highly efficient isolation of native ERK signaling complexes, which are suitable for subsequent analysis by mass spectrometry...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924549/identification-of-ochratoxin-a-producing-black-aspergilli-from-grapes-using-loop-mediated-isothermal-amplification-lamp-assays
#8
Michelangelo Storari, Giovanni A L Broggini
The loop-mediated isothermal amplification (LAMP) allows the rapid and specific amplification of target DNA under isothermal conditions without a prior DNA purification step. Moreover, successful amplifications can be directly evaluated through a color change of the reaction solutions. Here, we describe two LAMP assays for the detection of ochratoxin-A producing black aspergilli isolated from grapes. The two assays can detect DNA of OTA-producing black aspergilli following a very simple sample preparation and have the potential to significantly speed up the routine monitoring of these toxigenic molds in vineyards...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27924330/access-to-functionalized-thienopyridines-via-a-reagent-capsule-assisted-coupling-thiolation-and-cyclization-cascade-sequence
#9
Jialing Cai, Shuo Huang, Ruenfa He, Lu Chen, Donghan Chen, Shaohua Jiang, Bin Li, Yibiao Li
Thienopyridines and related heterocycles were prepared in a straightforward manner in moderate to good yields and under mild conditions by palladium-catalysed cross-coupling of ortho-fluorinated iodopyridines and terminal alkynes, followed by a reagent-capsule-assisted thiolation cyclization process. By applying paraffin wax capsules to prevent catalyst poisoning and undesired side reactions, the separation and purification processes were reduced.
December 7, 2016: Organic & Biomolecular Chemistry
https://www.readbyqxmd.com/read/27924296/data-in-support-of-covalent-attachment-of-tyrosinase-onto-cyanuric-chloride-crosslinked-magnetic-nanoparticles
#10
Kourosh Abdollahi, Farshad Yazdani, Reza Panahi
Preparation and characterization of cross linked amine-functionalized magnetic nanoparticles as an appropriate support for covalent immobilization on tyrosinase was presented in the study "Covalent immobilization of tyrosinase onto cyanuric chloride crosslinked amine-functionalized superparamagnetic nanoparticles: synthesis and characterization of the recyclable nanobiocatalyst" (Abdollahi et al., 2016 ) [1]. Herein, complementary data regarding X-ray powder diffraction (XRD) to characterize the synthesized magnetic nanoparticles, and transmission electron microscopy (TEM) to determine the size and morphology of tyrosinase immobilized magnetic nanoparticles (tyrosinase-MNPs) were reported...
December 2016: Data in Brief
https://www.readbyqxmd.com/read/27923737/halotag%C3%A2-evaluation-of-a-covalent-one-step-immobilization-for-biocatalysis
#11
Johannes Döbber, Martina Pohl
Easy, fast and gentle immobilization for the efficient reuse of important biocatalysts is highly demanded. We used the commercially available HaloTag™ technology (Promega), so far relatively unknown in the context of biocatalysis, to immobilize the benzaldehyde lyase from P. fluorescence (PfBAL). Immobilization mediated by this fusion tag proceeds rapidly within minutes from crude extracts yielding covalently attached enzymes in high purity, making expensive and laborious previous chromatographic purification steps obsolete, which strongly reduces the costs for biocatalyst immobilization...
December 3, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27923358/acthaside-a-new-chromone-derivative-from-acacia-ataxacantha-and-its-biological-activities
#12
Abdou Madjid O Amoussa, Mélanie Bourjot, Latifou Lagnika, Catherine Vonthron-Sénécheau, Ambaliou Sanni
BACKGROUND: Acacia ataxacantha (Fabaceae), used in traditional medicine grows in the South-West of Bénin. Ethyl acetate extract of the barks of this species was previously reported to display various bioactivities, including antibacterial, antifungal and antioxidant activities. In the present study, we investigate the antimicrobial and antioxidant activities of compound isolated from ethyl acetate extract of Acacia ataxacantha. METHODS: Purification, isolation and structural identification of isolated compound were done using various chromatographic and spectroscopic methods...
December 7, 2016: BMC Complementary and Alternative Medicine
https://www.readbyqxmd.com/read/27923319/non-invasive-swansea-criteria-are-valuable-alternatives-for-diagnosing-acute-fatty-liver-of-pregnancy-in-a-chinese-population
#13
Shan Wang, Shan-Ling Li, Yan-Xia Cao, Yan-Ping Li, Jin-Lai Meng, Xie-Tong Wang
BACKGROUND: This study aims to assess the diagnostic and prognostic value of Swansea criteria in diagnosing acute fatty liver of pregnancy (AFLP) in a Chinese population. METHODS: A retrospective study was conducted on 52 Chinese women diagnosed with AFLP. All selected cases were reassessed using the Swansea criteria with special focus on the non-invasive criteria, since performing a liver biopsy for this indication is rare in a Chinese population. RESULTS: Ninety point four percent of patients fulfilled five or more of the Swansea criteria...
December 6, 2016: Journal of Maternal-fetal & Neonatal Medicine
https://www.readbyqxmd.com/read/27920885/cloning-and-expression-of-soluble-recombinant-hiv-1-crf35-protease-hp-thioredoxin-fusion-protein
#14
Asaad Azarnezhad, Zohreh Sharifi, Rahmatollah Seyedabadi, Arshad Hosseini, Behrooz Johari, Mahsa Sobhani Fard
BACKGROUND: As a drug target and an antigenic agent, HIV-1 protease (HIV-1 PR) is at the center of attention for designing anti-AIDS inhibitors and diagnostic tests. In previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in E. coli and investigation of its immunoreactivity. METHODS: Protease coding region was isolated from the serum of an infected individual, amplified by RT-PCR and cloned into PTZ57R using TA-cloning...
October 2016: Avicenna Journal of Medical Biotechnology
https://www.readbyqxmd.com/read/27920826/expression-and-purification-of-truncated-diphtheria-toxin-dt386-in-escherichia-coli-an-attempt-for-production-of-a-new-vaccine-against-diphtheria
#15
Fatemeh Shafiee, Mohammad Rabbani, Mahdi Behdani, Ali Jahanian-Najafabadi
The aim of this study was to produce a recombinant protein consisting of the catalytic and translocation domains of diphtheria toxin for its later application as a vaccine candidate against Corynebacterium diphtheria. To achieve this goal, at first, the amino acid sequence of DT386 was used for prediction of T and B cell epitopes using on-line servers. The DT386 coding sequence was synthesized and subcloned into the NcoI and XhoI sites of pET28a plasmid and recombinant pET28a plasmid was used to transform Escherichia coli BL21 (DE3) host cells...
October 2016: Research in Pharmaceutical Sciences
https://www.readbyqxmd.com/read/27920276/studies-of-recombinant-twa1-reveal-constitutive-dimerisation
#16
Ore Francis, Genevieve E Baker, Paul R Race, Josephine C Adams
The mammalian muskelin/RanBP9/CTLH complex and the S. cerevisiae GID complex are large, multi-protein complexes that each contain a RING E3 ubiquitin ligase. The yeast GID complex acts to degrade a key enzyme of gluconeogenesis, fructose 1,6-bisphosphatase, under conditions of abundant fermentable carbon sources. However, the assembly and functions of the mammalian complex remain poorly understood. A striking feature of these complexes is the presence of multiple proteins that contain contiguous lissencephaly-1 homology (LisH), C-terminal to LisH (CTLH) and C-terminal CT11-RanBP9 (CRA) domains...
December 5, 2016: Bioscience Reports
https://www.readbyqxmd.com/read/27920166/revealing-the-heterogeneous-contamination-process-in-metal-nanoparticulate-catalysts-in-co-gas-without-purification-by-in-situ-environmental-transmission-electron-microscopy
#17
Tetsuya Uchiyama, Hideto Yoshida, Naoto Kamiuchi, Hideo Kohno, Seiji Takeda
It is known that samples become contaminated in CO gas of high pressure unless care is taken with the gas supply line to an environmental transmission electron microscope. This technical note reveals the heterogeneous formation process of contamination in situ on a nanoparticulate catalyst sample. It is shown that the surface of metal nanoparticles is preferentially contaminated, while the surface of metal oxide supports remains uncontaminated. It is also demonstrated that the contamination is suppressed by introducing a gas purifier in a gas supply line...
December 2016: Microscopy
https://www.readbyqxmd.com/read/27919837/ex-vivo-itreg-differentiation-revisited-convenient-alternatives-to-existing-strategies
#18
Billur Akkaya, Amanda H Holstein, Christopher Isaac, Mitra P Maz, Deborah D Glass, Ethan M Shevach, Munir Akkaya
Ex-vivo differentiation of regulatory T cells (Tregs) from naïve CD4(+) T-cells has been widely used in immunological research. Isolation of a highly pure naïve T cell population is the key factor that determines the efficiency of subsequent Treg differentiation. Currently, this step relies mostly on FACS sorting, which is often costly, time consuming, and inconvenient. Alternatively, magnetic separation of T-cells can be performed; yet, available protocols fail to reach sort level purity and consequently result in low Treg differentiation efficiency...
December 2, 2016: Journal of Immunological Methods
https://www.readbyqxmd.com/read/27919517/centrifugal-partition-chromatography-a-preparative-tool-for-isolation-and-purification-of-xylindein-from-chlorociboria-aeruginosa
#19
Anukul Boonloed, Genevieve L Weber, Kelly M Ramzy, Veronica R Dias, Vincent T Remcho
A centrifugal partition chromatography (CPC) method was developed for the preparative-scale isolation and purification of xylindein from the wood-staining fungi, Chlorociboria aeruginosa. Xylindein, a blue-green pigment naturally secreted from the hyphae and fruiting bodies of the fungus, has great value in the decorative wood industry and textile coloration. Xylindein has great potential for use as a fluorescent labeling agent as well as in organic semiconductor applications. However, a primary limitation of xylindein is its poor solubility in most common HPLC solvents...
November 18, 2016: Journal of Chromatography. A
https://www.readbyqxmd.com/read/27918991/sample-preparation-of-serum-to-allow-capillary-electrophoresis-analysis-of-prostate-specific-antigen-isoforms
#20
Noemi Farina-Gomez, Silvia Barrabes, Jorge E Gomez-Lopez, Monica Gonzalez, Angel Puerta, Diana Navarro-Calderon, Eduardo Albers-Acosta, Carlos Olivier, Jose C Diez-Masa, Rosa Peracaula, Mercedes de Frutos
Prostate cancer is the second most frequently diagnosed cancer in men worldwide. Currently prostate specific antigen (PSA) serum concentration is the most used prostate cancer marker, but it only shows limited specificity. Because PSA glycosylation is altered by prostate cancer, detecting glycosylation changes could increase PSA specificity as a prostate cancer marker. Changes in PSA glycosylation can modify its electrophoretic- behavior and techniques such as capillary zone electrophoresis (CZE) and two-dimensional electrophoresis (2-DE) could be applied to detect changes in PSA glycosylation...
November 24, 2016: Journal of Pharmaceutical and Biomedical Analysis
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