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Anas Alshishani, Salizawati Muhamad Salhimi, Bahruddin Saad
A new salting-out assisted liquid-liquid extraction (SALLE) sample preparation method for the determination of the polar anti-diabetic biguanide drugs (metformin, buformin and phenformin) in blood plasma, urine and lake water samples were developed. The SALLE was performed by mixing samples (plasma (0.2mL), urine or lake water (1.0mL)) with acetonitrile (0.4mL for plasma, 0.5mL for urine or lake water), sodium hydroxide powder was then added for the phase separation. The effects of type of salting-out reagent, type of extraction solvent, volumes of acetonitrile and sample, amount of sodium hydroxide, vortexing and centrifugation times on the extraction efficiency were investigated...
December 9, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Swatantra Pratap Singh, Yilun Li, Jibo Zhang, James M Tour, Christopher J Arnusch
Graphene based materials have profoundly impacted research in nanotechnology and this has significantly advanced biomedical, electronics, energy, and environmental applications. Laser-induced graphene (LIG) is made photothermally and has enabled a rapid route for graphene layers on polyimide surfaces. However, polysulfone (PSU), polyethersulfone (PES), and polyphenylsulfone (PPSU) are highly used in numerous applications including medical, energy and water treatment and they are critical components of polymer membranes...
December 14, 2017: ACS Nano
Rodrigo Vazquez-Lombardi, Damien Nevoltris, Ansha Luthra, Peter Schofield, Carsten Zimmermann, Daniel Christ
Recombinant expression of antibody molecules in mammalian cells offers important advantages over traditionally utilized bacterial expression, including glycosylation required for antibody functionality and markedly reduced levels of endotoxin contamination. Advances in transient mammalian expression systems enable high yields (>100 mg/liter) that now allow for effective recombinant antibody production at a reasonable cost. Here, we provide step-by-step protocols for the design and recombinant expression of full-length IgG antibodies and antibody-derived constructs (including Fab, Fc-fusions and bispecifics) in mammalian cells...
January 2018: Nature Protocols
Jake Pawlowski, Ian Carrick, Igor A Kaltashov
Profiling of complex proteins by means of mass spectrometry (MS) frequently requires that certain chemical modifications of their covalent structure (e.g., reduction of disulfide bonds) be carried out prior to the MS or MS/MS analysis. Traditionally, these chemical reactions take place in the off-line mode to allow the excess reagents (the majority of which interfere with the MS measurements and degrade the analytical signal) to be removed from the protein solution prior to MS measurements. In addition to a significant increase in the analysis time, chemical reactions may result in a partial or full loss of the protein if the modifications adversely affect its stability, e...
December 14, 2017: Analytical Chemistry
Molly M MacInnes, Sofiya Hlynchuk, Saurabh Acharya, Nicolai Lehnert, Stephen Maldonado
Reduced graphene oxide (RGO) films have been prepared by immersion of graphene oxide (GO) films at room temperature in non-aqueous solutions containing simple, outer-sphere metallocene reductants. Specifically, solutions of cobaltocene, cobaltocene & trifluoroacetic acid, and decamethylcobaltocene each showed activity for the rapid reduction of GO films cast on a wide variety of substrates. Each reactant increased the conductivity of the films by several orders of magnitude, with RGO films prepared with either decamethylcobaltocene or with cobaltocene & trifluoroacetic acid possessing the highest conductivities (~ 104 S m-1)...
December 14, 2017: ACS Applied Materials & Interfaces
Yue Zhao, Guowen Liu, Suk Kwok, Barry R Jones, Jane Liu, David Marchisin, Philip E Joyce, Jon Peterson, Jim X Shen
AIM: Recombinant FGF21 analogs are under wide ranging investigations as a potential therapeutic agent for Type 2 diabetes, as well as other metabolic disorders. The endogenous FGF21 is often used as a surrogate pharmacodynamic(PD) biomarker to assess drug efficacy and safety. Results/methodology: Immunocapture was performed using a monoclonal antibody which had been generated to bind to specific domain of native FGF21 as the capture reagent. After immunocapture, enzymatic digestion was performed and a native FGF21-specific tryptic peptide was monitored using LC-MS/MS by selective reaction monitoring...
December 14, 2017: Bioanalysis
Seetharaman Manojveer, Sebastian Forrest, Magnus T Johnson
Substituted arylethanols can be coupled using a readily available Ru-catalyst in a fully deoxygenative manner to produce hydrocarbon chains in one step. Control experiments indicate that the first deoxygenation occurs through an aldol condensation whereas the second occurs through a base-induced net decarbonylation. This double deoxygenation enables further development in the use of alcohols as versatile and green alkylating reagents, as well as in other fields such as deoxygenation and upgrading of overfunctionalized biomass to produce hydrocarbons...
December 14, 2017: Chemistry: a European Journal
Avishek Roy, Mrinal Kanti Das, Saikat Chaudhuri, Alakesh Bisai
We report an efficient direct alkynylations of 3-alkyl/aryl 2-oxindoles employing Waser's ethynyl-1,2-BenziodoXol-3(1H)-one (EBX) to afford a wide variety of 3-alkynyl-3-alkyl/aryl 2-oxindole under transition-metal free condition. In addition to activated carbonyl compounds viz. 2-oxindole-3-alkylcarboxylates, this direct alkynylations protocol works efficiently on 3-alkyl/aryl 2-oxindols as well thereby widening the scope even further. Eventually, a Pd(0)-catalyzed asymmetric decarboxylative allylation of few products is shown to furnish synthetically viable enantioenriched 2-oxindoles with C-3 quaternary stereocenters...
December 14, 2017: Journal of Organic Chemistry
Nao Hiwatashi, Iv Kraja, Peter A Benedict, Gregory R Dion, Renjie Bing, Bernard Rousseau, Milan R Amin, Danielle M Nalband, Kent Kirshenbaum, Ryan C Branski
OBJECTIVES/HYPOTHESIS: Our laboratory and others hypothesized that Smad3 is a principle mediator of the fibrotic phenotype in the vocal folds (VFs), and we further posited that alteration of Smad3 expression through short interfering (si)RNA holds therapeutic promise, yet delivery remains challenging. To address this issue, we employed a novel synthetic oligomer, lipitoid, complexed with siRNA to improve stability and cellular uptake with the goal of increased efficiency of RNA-based therapeutics...
December 14, 2017: Laryngoscope
Ilich A Ibarra, Alejandro Islas-Jácome, Eduardo González-Zamora
Polyheterocycles are one of the most desired synthetic targets due to their numerous and valuable applications in various fields. Multicomponent reactions (MCRs) are highly convergent one-pot processes, in which three or more reagents are combined sequentially to construct complex products, with almost all the atoms coming from the starting reagents. In this context, the syntheses of 'heterocycles' via MCR-based processes have been reviewed a number of times. However, there is not a single review (recent or otherwise) covering the synthesis of 'polyheterocycles' via a direct MCR or via a one-pot process involving MCRs coupled to further cyclizations (via ionic, metal-catalyzed, pericyclic, or free-radical-mediated cyclizations)...
December 13, 2017: Organic & Biomolecular Chemistry
S S Kappala, M Alessandrini, T Matlhako, E Beltchev, R Pool, M S Pepper
Acute myeloid leukemia (AML) is characterized by proliferation of the myeloid lineage and accumulation of immature hematopoietic cells in the bone marrow and is typified by marked heterogeneity both in response to treatment and survival. AMLprofiler is a qualitative in vitro diagnostic microarray incorporating seven molecular biomarkers used to diagnose and predict posttherapy survival rates. In this study, we compared AMLprofiler to routine AML diagnostic methodologies employed in South Africa, focusing on consistency of the results, cost, and time to result...
2017: Stem Cells International
Nagatoshi Nishiwaki
An efficient synthetic protocol based on a new concept named "the pseudo-intramolecular process" is developed. Substrates are brought closed to each other by forming a salt, and this spatial proximity facilitates an efficient reaction like an intramolecular process, despite actually proceeding via an intermolecular pathway. This concept can be widely applied in organic synthesis. For instance, α-aryl-β-keto esters undergo a transacylation with amines accompanied by high efficiency, regioselectivity, and chemoselectivity...
December 14, 2017: Journal of Oleo Science
Joseph D Dougherty
Translating ribosome affinity purification is a method initially developed for profiling mRNA from genetically defined cell types in complex tissues. It has been applied both to identify target molecules in cell types that are important for controlling a variety of behaviors in the brain, and to understand the molecular consequences on those cells due to experimental manipulations, ranging from drugs of abuse to disease-causing mutations. Since its inception, a variety of methodological advances are opening new avenues of investigation...
December 13, 2017: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Carla Surlis, Keelan McNamara, Eoin O'Hara, Sinead Waters, Marijke Beltman, Joseph Cassidy, David Kenny
BACKGROUND: Caesarean section is a routine veterinary obstetrical procedure employed to alleviate dystocia in cattle. However, CS, particularly before the onset of labour, is known to negatively affect neonatal respiration and metabolic adaptation in humans, though there is little published information for cattle. The aim of this study was to investigate the effect of elective caesarean section (ECS) or normal trans-vaginal (TV) delivery, on lung and jejunal gene expression profiles of neonatal calves...
December 14, 2017: BMC Veterinary Research
Huang Jianfeng, Wang Yueqian, Wang Lunan, Pan Yang, Chen Tong
PURPOSE: To investigate the effect of microRNA-16 on hypoxia-induced VEGF expression of ARPE-19 cells Methods: ARPE-19 cells were cultivated under normoxia and hypoxia state. At 0h, 12h, 24h and 48h after cultivation, the supernate of the culture medium were separated to test the VEGF secretion by ELISA, and the cells were purified to measure the expression of VEGF mRNA and microRNA-16 by qRT-PCR; microRNA-16 mimic was then transfected into ARPE-19 cells by the Hiperfect transfection reagent, a liposome transfection system...
December 13, 2017: Cutaneous and Ocular Toxicology
Gour Chand Daskhan, Hanh-Thuc Ton Tran, Peter Meloncelli, Todd L Lowary, Lori J West, Christopher W Cairo
The design and synthesis of multivalent ligands displaying complex oligosaccharides is necessary for the development of therapeutics, diagnostics, and research tools. Here, we report an efficient conjugation strategy to prepare complex glycoconjugates with four copies of one or two separate glycan epitopes, providing 4-8 carbohydrate residues on a tetravalent poly(ethylene glycol) scaffold. This strategy provides complex glycoconjugates that approach the size of glycoproteins (15-18 kDa) while remaining well-defined...
December 13, 2017: Bioconjugate Chemistry
D Robert Sutherland, Andrea Illingworth, Iuri Marinov, Fernando Ortiz, John Andreasen, Dan Payne, Paul K Wallace, Michael Keeney
Since publication in 2010 of the International Clinical Cytometry Society (ICCS) Consensus Guidelines for detection of Paroxysmal nocturnal hemoglobinuria (PNH) by flow cytometery, a great deal of work has been performed to develop, optimize and validate a number of high-sensitivity assays to detect PNH phenotypes in both Red Blood Cells (RBCs) and White Blood Cells (WBCs, neutrophils and monocytes). This section (Part 2) of the updated ICCS PNH Consensus Guidelines will focus on specific instrument setup for these PNH assays, the identification and proper testing of appropriate antibody conjugates and combinations therof, and basic assay design...
December 13, 2017: Cytometry. Part B, Clinical Cytometry
Lijun Zheng, Wenhui Wang, Jinjin Liu, Xuhui Chen, Sanjing Li, Qiaoli Wang, Yuqi Huo, Chuan Qin, Shuo Shen, Mingchen Wang
Noroviruses (NoVs) are increasingly recognized as the leading cause of acute non-bacterial gastroenteritis worldwide. To screen for NoV-specific monoclonal antibodies (mAbs) with wide spectrum binding activities that could be used for the development of NoV-related detection reagents, we immunized mice with a combination of virus like particles (VLPs) derived from eight different genotypes (two from genogroup I and six from genogroup II), of which two (GI.7 and GII.2) were newly produced VLPs. Indirect enzyme-linked immunosorbent assay (ELISA) confirmed that two mAbs (8D8 and 10B11) bound to all eight major capsid proteins (VP1) with varied binding abilities...
December 13, 2017: Journal of Medical Virology
Pin-Wen Huang, Cong-Zhi Wang, Qun-Yan Wu, Jian-Hui Lan, Gang Song, Zhi-Fang Chai, Wei-Qun Shi
Separation of trivalent actinides (An(iii)) and lanthanides (Ln(iii)) in spent nuclear fuel reprocessing is extremely challenging mainly owing to their similar chemical properties. Two amine-type reagents, tetrakis(2-pyridyl-methyl)-1,2-ethylenediamine (TPEN) and its hydrophobic derivative N,N,N',N'-tetrakis((4-butoxypyridin-2-yl)methyl)-ethylenediamine (TBPEN), have been identified to possess a selectivity for Am(iii) over Eu(iii). In this work, the structures, bonding nature, and thermodynamic behaviors of the Am(iii) and Eu(iii) complexes with these two ligands have been systematically studied via scalar relativistic density functional theory (DFT) calculations...
December 13, 2017: Physical Chemistry Chemical Physics: PCCP
Shaohua Chen, Yan Sun, Jia Li, Ling Xiong
Amyl xanthate (AX), a widely used reagent in sulfide mineral flotation, has caused serious environmental problems. Manganese dioxides (MnO2), ubiquitous in natural soils and sediments, are one kind of important oxidant or catalyst in facilitating organic pollutant decomposition. In this study, the oxidative degradation of AX by MnO2 was investigated in batch experiments. It was found that up to 90% AX was decomposed by MnO2 in 30 min and the removal efficiency of AX depended strongly on the loading of MnO2 and pH value in the solution where the reactions closely followed a pseudo-first-order model...
December 2017: Water Science and Technology: a Journal of the International Association on Water Pollution Research
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