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https://www.readbyqxmd.com/read/28109042/expression-and-characterization-of-a-codon-optimized-blood-coagulation-factor-viii
#1
S A Shestopal, J-J Hao, E Karnaukhova, Y Liang, M V Ovanesov, M Lin, J H Kurasawa, T K Lee, J H McVey, A G Sarafanov
BACKGROUND: Production of recombinant factor VIII (FVIII) is challenging due to its low expression. It was previously shown that codon-optimization of a B domain-deleted FVIII (BDD-FVIII) cDNA resulted in increased protein expression. However, it is well-recognized that synonymous mutations may affect the protein structure and function. OBJECTIVES: To compare biochemical properties of a BDD-FVIII expressed from codon-optimized (CO) and the wild-type (WT) cDNAs. METHODS: Each variant of the BDD-FVIII was expressed in several independent CHO cell lines, generated using a lentiviral platform...
January 21, 2017: Journal of Thrombosis and Haemostasis: JTH
https://www.readbyqxmd.com/read/28108388/c-difficile-pcr-ribotype-023-might-go-undetected-when-using-chromid-c-difficile-agar
#2
E Reigadas, L Alcalá, M Marín, A Martín, E Bouza
We compared the performance of the new chromogenic medium ChromID C. difficile with that of CLO agar. ChromID C. difficile agar is a sensitive medium that can accelerate the presumptive identification of C. difficile, however ribotype 023 might go undetected when using this chromogenic medium.
January 17, 2017: Anaerobe
https://www.readbyqxmd.com/read/28108122/-fast-methods-of-fungal-and-bacterial-identification-maldi-tof-mass-spectrometry-chromogenic-media
#3
María Siller-Ruiz, Sara Hernández-Egido, Fernando Sánchez-Juanes, José Manuel González-Buitrago, Juan Luis Muñoz-Bellido
MALDI-TOF mass spectrometry is now a routine resource in Clinical Microbiology, because of its speed and reliability in the identification of microorganisms. Its performance in the identification of bacteria and yeasts is perfectly contrasted. The identification of mycobacteria and moulds is more complex, due to the heterogeneity of spectra within each species. The methodology is somewhat more complex, and expanding the size of species libraries, and the number of spectra of each species, will be crucial to achieve greater efficiency...
January 17, 2017: Enfermedades Infecciosas y Microbiología Clínica
https://www.readbyqxmd.com/read/28105172/her2-gene-amplification-in-patients-with-prostate-cancer-evaluating-a-cish-based-method
#4
Nazanin Sharifi, Arash Salmaninejad, Samira Ferdosi, Abolfazl Nesaei Bajestani, Malihe Khaleghiyan, Mehrdad Asghari Estiar, Mansour Jamali, Mohammad Reza Nowroozi, Abbas Shakoori
Prostate cancer (PCa) is one of the most widespread malignancies in the world. The role of the human epidermal growth factor receptor 2 (HER2) in the pathogenesis and progression of human PCa remains poorly understood. In contradiction with breast cancer, studies on HER2 overexpression and gene amplification in PCa have produced varying results, although the HER2 oncogene has been implicated in the biology of numerous tumor types, and serves as a prognostic marker and therapeutic target in breast cancer. Technical challenges are considered the main reasons for data discrepancies...
December 2016: Oncology Letters
https://www.readbyqxmd.com/read/28103697/3-4-hydroxyphenyl-propionic-acid-the-forgotten-detection-substrate-for-ligand-binding-assay-based-bioanalysis
#5
Gregor Jordan, Kay-Gunnar Stubenrauch, Julia Heinrich, Roland F Staack
Ligand-binding assays are ideal for routine bioanalysis, but we reason that the straightforward replacement of the conventional chromogenic horseradish peroxidase substrate, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid, of a routinely used preclinical immunoassay to detect hIgG, with the fluorogenic 3-(4-hydroxyphenyl)propionic acid would broaden the narrow dynamic range. The replacement leads to a sensitivity of 0.47 (minimum required dilution [MRD] 10) and 1.02 (MRD 50) ng/ml, and dynamic ranges of 3...
January 20, 2017: Bioanalysis
https://www.readbyqxmd.com/read/28092096/associations-between-pre-eclampsia-and-protein-c-and-protein%C3%A2-s-levels-among-pregnant-nigerian-women
#6
Helen C Okoye, Lisa I Eweputanna, Anthony O U Okpani, Oseikhuemen A Ejele
OBJECTIVE: To evaluate levels of protein C and free protein S among women with pre-eclampsia, and determine whether there is a relationship between deficiencies and pre-eclampsia. METHODS: A cross-sectional study was conducted at a hospital in Nigeria from July 2013 to March 2014 among 90 pregnant women with pre-eclampsia (blood pressure ≥140/90 mm Hg, proteinuria ≥300 mg in 24 hours) and 90 normotensive pregnant women (control group). Plasma levels of protein C and free protein S were analyzed by enzyme-linked immunosorbent assay, and protein C activity by a chromogenic method...
December 16, 2016: International Journal of Gynaecology and Obstetrics
https://www.readbyqxmd.com/read/28089737/browplasminin-a-condensed-tannin-with-anti-plasmin-activity-isolated-from-an-aqueous-extract-of-brownea-grandiceps-jacq-flowers
#7
Betzabeth Pereira, Josmary Brazón, Mónica Rincón, Eva Vonasek
ETHNOPHARMACOLOGICAL RELEVANCE: Following Venezuelan traditional medicine, females with heavy menstrual blood loss (menorrhagia) drink Brownea grandiceps Jacq. flowers (BG) decoctions to reduce the bleeding. In a previous study, we demonstrated that BG aqueous extract (E) possesses a potent anti-fibrinolytic activity capable of inhibiting plasmin, the main serine-protease that degrades fibrin. It is widely known that plasmin inhibitors are often used as anti-fibrinolytics to reduce bleeding during surgeries with high risk of blood loss such as cardiac, liver, vascular, tooth extraction and large orthopedic procedures, as well as for menorrhagia treatments...
January 12, 2017: Journal of Ethnopharmacology
https://www.readbyqxmd.com/read/28089408/the-various-assays-for-measuring-activity-states-of-factor-viia-in-plasma-and-therapeutic-products-diagnostic-value-and-analytical-usefulness-in-various-pathophysiological-states
#8
REVIEW
Jean Amiral, Claire Dunois, Cédric Amiral, Jerard Seghatchian
The key coagulation factor FVII, and its activated form FVIIa, present a major interest for their role at the initiation phase of blood coagulation, and because they can activate all blood coagulation cascade, through the extrinsic, but also the intrinsic pathway. Blood activation initiated through FVII is first presented, as it is understood nowadays. Measurement of FVII and FVIIa were of main interest for epidemiological studies, but FVIIa contribution to assay results was only deduced. The introduction of specific FVIIa assays, functional or immunoassays, allowed measuring directly FVIIa without any interference of non-activated FVII, or other coagulation factors or their activated forms...
December 29, 2016: Transfusion and Apheresis Science
https://www.readbyqxmd.com/read/28087107/establishment-of-the-2nd-korean-national-biological-reference-standard-for-blood-coagulation-factor-viii-c-concentrate
#9
Naery Lee, Ji Suk Seo, Jae Ok Kim, Sang Ja Ban
Since the 1st Korean national biological reference standard for factor (F)VIII concentrate, established in 2001, has shown declining potency, we conducted this study to replace this standard with a 2nd Korean national biological reference standard for blood coagulation FVIII concentrate. The candidate materials for the 2nd standard were prepared in 8000 vials with 10 IU/ml of target potency, according to the approved manufacturing process of blood coagulation Factor VIII:C Monoclonal Antibody-purified, Freeze-dried Human Blood Coagulation Factor VIII:C...
January 10, 2017: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/28073895/evaluation-of-genotypic-and-phenotypic-methods-to-detect-carbapenemase-production-in-gram-negative-bacilli
#10
Allison R McMullen, Melanie L Yarbrough, Meghan A Wallace, Angela Shupe, Carey-Ann D Burnham
BACKGROUND: Carbapenemase-producing gram-negative bacteria (CP-GNB) are an urgent and expanding public health threat. Rapid and accurate identification of these organisms facilitates infection prevention efforts in healthcare facilities. The objective of our study was to evaluate methods to detect and identify CP-GNB. METHODS: We examined 189 carbapenem-resistant GNB (CR-GNB), including Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii complex, using 3 different methods: 2 methods to screen isolates of GNB for carbapenemase production [the carbapenem inactivation method (CIM) and 2 chromogenic agars] and a molecular method (Cepheid GeneXpert Carba-R) to identify the mechanism of carbapenem resistance and the associated resistance genes (blaKPC, blaNDM, blaIMP, blaOXA-48-like, and blaVIM)...
January 10, 2017: Clinical Chemistry
https://www.readbyqxmd.com/read/28069655/mutation-in-an-unannotated-protein-confers-carbapenem-resistance-in-mycobacterium-tuberculosis
#11
Pankaj Kumar, Amit Kaushik, Drew Bell, Varsha Chauhan, Fangfang Xia, Rick L Stevens, Gyanu Lamichhane
β-lactams are the most widely used antibacterials. Among β-lactams, carbapenems are considered the last line of defense against recalcitrant infections. As recent developments have prompted consideration of carbapenems for treatment of drug-resistant tuberculosis, it is only a matter of time before Mycobacterium tuberculosis (Mtb) strains resistant to these drugs will emerge. Here, we investigated the genetic basis that confers such resistance. To our surprise, instead of mutations in the known β-lactam targets, a single nucleotide polymorphism in the Rv2421c-Rv2422 intergenic region was common among Mtb mutants selected with meropenem or biapenem...
January 9, 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/28067433/plasmonic-imaging-of-the-interfacial-potential-distribution-on-bipolar-electrodes
#12
Meisam Hasheminejad, Yimin Fang, Meng Li, Yingyan Jiang, Wei Wang, Hong-Yuan Chen
Bipolar electrochemistry is based on the gradient distribution of free-electron density along an electrically isolated electrode, which causes a positive electrode potential at one end and a negative potential at the other, allowing for wide applications in analytical chemistry and materials science. To take full advantage of its wireless and high-throughput features, various types of optical probes, such as pH indicators and fluorescence and electrochemiluminescence reagents, have often been used to indirectly monitor the interfacial electron transfer through chromogenic or fluorogenic reactions...
January 9, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28064137/a-highly-selective-chromogenic-sensor-for-mn-2-turn-off-fluorometric-for-hg-2-ion-and-turn-on-fluorogenic-sensor-for-f-ion-with-the-practical-application
#13
Neha Gupta, Divya Singhal, Ashok Kumar Singh, Neetu Singh, U P Singh
A colorimetric and fluorometric probe (E)-2-((8-hydroxy-1,2,3,5,6,7-hexahydropyrido[3,2,1-ij]quinolin-9-yl)methylene)hydrazinecarbothioamide based on thiosemicarbazide and julolidine moieties has been synthesised in pure crystalline form and characterized by (1)H NMR, UV-vis, elemental analysis and single crystal XRD. The probe functioned as multitarget ion sensor, detect biologically important metal ions Hg(2+) and Mn(2+) in dual channel mode. Meanwhile, in mixed solvent media DMF/H2O [8:2], probe displayed selectivity for Hg(2+) over other cations by the emission spectrum...
January 4, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/28060344/visualizing-the-interrenal-steroidogenic-tissue-and-its-vascular-microenvironment-in-zebrafish
#14
Chih-Wei Chou, Jamie Lin, Hsin-Yu Hou, Yi-Wen Liu
This protocol introduces how to detect differentiated interrenal steroidogenic cells through a simple whole-mount enzymatic activity assay. Identifying differentiated steroidogenic tissues through chromogenic histochemical staining of 3-β-Hydroxysteroid dehydrogenase /Δ(5-4) isomerase (3β-Hsd) activity-positive cells is critical for monitoring the morphology and differentiation of adrenocortical and interrenal tissues in mammals and teleosts, respectively. In the zebrafish model, the optical transparency and tissue permeability of the developing embryos and larvae allow for whole-mount staining of 3β-Hsd activity...
December 21, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28052306/laboratory-monitoring-of-parenteral-direct-thrombin-inhibitors
#15
Elizabeth M Van Cott, A Joshua Roberts, William E Dager
Argatroban and bivalirudin are parenteral direct inhibitors of the activity of thrombin, but, unlike heparin, can inhibit both soluble as well as clot-bound thrombin. These agents do not require antithrombin as a cofactor for activity. The parenteral direct thrombin inhibitors (DTIs) can be used in a variety of settings, including heparin-induced thrombocytopenia (HIT) or an allergy to heparin, and patients requiring anticoagulation for an invasive cardiovascular intervention. Both agents have a relatively short half-life in patients without organ system failure and are typically administered by continuous infusion...
January 4, 2017: Seminars in Thrombosis and Hemostasis
https://www.readbyqxmd.com/read/28036168/development-of-an-azoreductase-based-reporter-system-with-synthetic-fluorogenic-substrates
#16
Narae Shin, Kenjiro Hanaoka, Wen Piao, Takuya Miyakawa, Tomotsumi Fujisawa, Satoshi Takeuchi, Shodai Takahashi, Toru Komatsu, Tasuku Ueno, Takuya Terai, Tahei Tahara, Masaru Tanokura, Tetsuo Nagano, Yasuteru Urano
Enzyme/substrate pairs, such as β-galactosidase with chromogenic x-gal substrate, are widely used as reporters to monitor biological events, but there is still a requirement for new reporter systems, which may be orthogonal to existing systems. Here, we focused on azoreductase (AzoR). We designed and synthesized a library of azo rhodamine derivatives as candidate fluo-rogenic substrates. These derivatives were non-fluorescent, probably due to ultrafast conformational change around the N=N bond after photoexcitation...
December 30, 2016: ACS Chemical Biology
https://www.readbyqxmd.com/read/28034905/absence-of-cytomegalovirus-in-glioblastoma-and-other-high-grade-gliomas-by-real-time-pcr-immunohistochemistry-and-in-situ-hybridization
#17
Matthias Holdhoff, Gunes Guner, Fausto J Rodriguez, Jessica L Hicks, Qizhi Zheng, Michael S Forman, Xiaobu Ye, Stuart A Grossman, Alan K Meeker, Christopher M Heaphy, Charles G Eberhart, Angelo M De Marzo, Ravit Arav-Boger
PURPOSE: Reports of cytomegalovirus (CMV) detection in high-grade gliomas (HGG)/glioblastoma (GBM) have been conflicting. We undertook a comprehensive approach to determine presence or absence of CMV in tissue, plasma and serum of HGG patients. EXPERIMENTAL DESIGN: In a retrospective arm, 25 fresh frozen tissues from GBM patients were tested for CMV by real-time PCR. Tissue microarrays from 70 HGG patients were tested by immunohistochemistry (IHC) and 20 formalin-fixed paraffin-embedded (FFPE) GBM tissues by IHC and chromogenic in situ hybridization (CISH), targeting CMV-encoded IE1/2 and pp65...
December 29, 2016: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
https://www.readbyqxmd.com/read/28028529/purification-of-lactoperoxidase-from-bovine-whey-and-investigation-of-kinetic-parameters
#18
Fatemeh Borzouee, Mohammad Reza Mofid, Jaleh Varshosaz, Seyed Ziyae Aldin Samsam Shariat
BACKGROUND: Lactoperoxidase (LPO) is related to mammalian peroxidase family which contains a wide spectrum of biological activities. Despite the wide studies on the LPO, there is little study has been performed to simplify and shorten the procedure of enzyme purification. The aim of this project was to purify the enzyme through a simple method, and investigating enzyme kinetic parameters. MATERIALS AND METHODS: LPO was purified from bovine whey through modified method of Yoshida (1990) using two steps of ammonium sulfate precipitation and ion-exchange chromatography...
2016: Advanced Biomedical Research
https://www.readbyqxmd.com/read/28013165/in-vitro-cytotoxicity-evaluation-of-graphene-oxide-from-the-peroxidase-like-activity-perspective
#19
Wei Zhang, Ying Sun, Zhichao Lou, Lina Song, Yang Wu, Ning Gu, Yu Zhang
In this study, PEGylated graphene oxide (PEG-GO)-hemin composite structure was constructed. Hemin in the form of nanoscaled aggregates were immobilized on PEG-GO sheets by the π-π stacking super-molecular interaction. Via catalyzing the oxidation of chromogenic substrates, we elicited the obtained PEG-GO-Hemin composite sheets have much higher peroxidase-like activity compared to hemin or PEG-GO alone, which is due to the introduction of enzyme active center of hemin with high dispersity, the excellent affinity to organic substrate through π-π stacking and/or electrostatic adsorption and the rapid electron transfer capability of PEG-GO...
December 19, 2016: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28013021/kanamycin-detection-based-on-the-catalytic-ability-enhancement-of-gold-nanoparticles
#20
Chengke Wang, Dan Chen, Qingqing Wang, Rong Tan
In this paper, we demonstrated that kanamycin could enhance the peroxidase-like activity of citrate-capped gold nanoparticles (AuNPs) through two steps: the attachment of kanamycin onto AuNPs through -NH2 (on kanamycin) and -COOH (on AuNPs) interactions; and the specifically interaction between glucoside on kanamycin and AuNPs which changes the surface property of AuNPs, and produced •OH radicals and Au(3+) in the solution, and catalyzed the chromogenic reactions between 3, 3', 5, 5'-tetramethylbenzidine (TMB) and H2O2...
December 18, 2016: Biosensors & Bioelectronics
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