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High Reproducibility (high + reproducibility)
Selected AbstractsRapid gas chromatography/mass spectrometry quinine determination in plasma after automated solid-phase extractionRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 17 2006Richard Damien The combined use of an automatic solid-phase extraction (SPE) apparatus with Oasis MCX cartridges and gas chromatography/mass spectrometry (GC/MS) to rapidly quantify quinine in biological samples with cyproheptadine as the internal standard is described. The selected ion monitoring mode, with the quantification ions m/z 136 and 287 (qualifier ions: m/z 261, 381 and 215, 96), allows the estimation of quinine levels, respectively. Separation was completed within 12.7,min. Excellent linearity was found up to 10 000,µg/L of plasma. The limit of detection (LOD) was 12.2,µg/L and the limit of quantification (LOQ) was 40.6,µg/L. High reproducibility (intra-assay CV range 1.9,4.3%, inter-assay CV range 2.2,11.3%) and accuracy values (intra-assay range 83.2,103.7%, inter-assay range 86.8,103.7%) were obtained. Recoveries were concentration-independent (97.2% and 89.8% for 4000 and 10 000,µg/L, respectively). This sensitive, simple assay for quinine in various matrices meets the current requirements for bioanalytical assays and may be used to monitor quinine levels in patients developing severe malaria with acute renal failure during hemofiltration. The optimal quinine dose in this situation is not really established and to improve clinical care, quinine concentrations might be explored to improve efficacy and minimise potential toxicity. Copyright © 2006 John Wiley & Sons, Ltd. [source] Use of automated microscopy for the detection of disseminated tumor cells in bone marrow samplesCYTOMETRY, Issue 4 2001Elin Borgen Abstract The use of automated microscopy has reached the maturity necessary for its routine use in the clinical pathology laboratory. In the following study we compared the performance of an automated microscope system (MDSÔ) with manual method for the detection and analysis of disseminated tumor cells present in bone marrow preparations from breast carcinoma patients. The MDS System detected rare disseminated tumor cells among bone marrow mononuclear cells with higher sensitivity than standard manual microscopy. Automated microscopy also proved to be a method of high reproducibility and precision, the advantage of which was clearly illustrated by problems of variability in manual screening. Accumulated results from two pathologists who had screened 120 clinical slides from breast cancer patients both by manual microscopy and by use of the MDS System revealed only two (3.8%) missed by the automatic procedure, whereas as many as 20 out of 52 positive samples (38%) were missed by manual screening. Cytometry (Comm. Clin. Cytometry) 46:215,221, 2001. © 2001 Wiley-Liss, Inc. [source] In vivo determination of muscle viscoelasticity in the human legACTA PHYSIOLOGICA, Issue 4 2001S. Fukashiro The purpose of this study was to examine the methodological validity of the free vibration technique for determining individual viscoelastic characteristics of the human triceps surae muscle-tendon complex (MTC) in vivo. Six subjects sat with first phalangeal joint of the forefoot on the edge of a force-plate. The special frame on the knee was loaded with weight (0,40 kg) for testing. Oscillations of the triceps surae MTC system were initiated with a hand-held hammer by tapping the weight. In order to keep the same posture, the output of the force plate was displayed on the oscilloscope and subjects were asked to maintain the beam on the oscilloscope at a particular location in relation to a reference line. The damped oscillations in conjunction with the equation of motion of a damped mass-spring model were used to calculate the viscosity of muscle (b) and the elasticity of muscle fibres and tendon (k) in each subject, considering moment arm of the ankle joint. With this arrangement, we have obtained high reproducibility in this method. The coefficient of variations (CVs) of b and k in five trials at each weight were quite small (range: 0.5,18.7% in b and 1.0,15.1% in k). There were no significant differences in viscoelastic coefficients between right and left legs. Therefore, it appears that free vibration technique, used here, is adequate in describing the viscoelastic characteristics of the triceps surae in vivo in humans. [source] Lead(II) Potentiometric Sensor Based on 1,4,8,11-Tetrathiacyclotetradecane Neutral Carrier and Lipophilic AdditivesELECTROANALYSIS, Issue 11 2008Mohamed Abstract A potentiometric sensor for lead(II) ions based on the use of 1,4,8,11-tetrathiacyclotetradecane (TTCTD) as a neutral ionophore and potassium tetrakis-(p -chlorophenyl)borate as a lipophilic additive in plasticized PVC membranes is developed. The sensor exhibits linear potentiometric response towards lead(II) ions over the concentration range of 1.0×10,5,1.0×10,2,mol L,1 with a Nernstian slope of 29.9,mV decade,1 and a lower limit of detection of 2.2×10,6,mol L,1 Pb(II) ions over the pH range of 3,6.5. Sensor membrane without a lipophilic additive displays poor response. The sensor shows high selectivity for Pb(II) over a wide variety of alkali, alkaline earth and transition metal ions. The sensor shows long life span, high reproducibility, fast response and long term stability. Validation of the method by measuring the lower limit of detection, lower limit of linear range, accuracy, precision and sensitivity reveals good performance characteristics of the proposed sensor. The developed sensor is successfully applied to direct determination of lead(II) in real samples. The sensor is also used as an indicator electrode for the potentiometric titration of Pb(II) with EDTA and potassium chromate. The results obtained agree fairly well with data obtained by AAS. [source] Disposable Gold Electrode Array for Simultaneous Electrochemical StudiesELECTROANALYSIS, Issue 1 2008Graciela Priano Abstract An efficient and inexpensive eight gold electrode array has been manufactured by a combination of screen printing and gold electrodeposition techniques. Gold electrodeposition was performed in potentiostatic and galvanostatic conditions. Different treatments, involving temperature and polishing control, led to electrodes with different roughness. The electrochemical behavior of the generated gold surface was studied by cyclic voltammetry showing the characteristic response of polycrystalline gold, in contrast with disposable gold electrodes fabricated by screen printing from gold inks. The electrodes were chemically modified through the adsorption of alkanethiols self-assembled monolayers and the coupling of a model protein. Both reactions were followed by cyclic voltammetry and Electrochemical Impedance Spectroscopy (EIS). The electrodes have shown high reproducibility in their electrochemical behavior as well as in their modifications. [source] Comparative metabolite profiling of carboxylic acids in rat urine by CE-ESI MS/MS through positively pre-charged and 2H-coded derivatizationELECTROPHORESIS, Issue 22 2008Wen-Chu Yang Abstract A new approach to the selective comparative metabolite profiling of carboxylic acids in rat urine was established using CE-MS and a method for positively pre-charged and 2H-coded derivatization. Novel derivatizing reagents, N -alkyl-4-aminomethyl-pyridinum iodide (alkyl=butyl, butyl-d9 or hexyl), containing quaternary amine and stable-isotope atoms (deuterium), were introduced for the derivatization of carboxylic acids. CE separation in positive polarity showed high reproducibility (0.99,1.32% RSD of migration time) and eliminated problems with capillary coating known in CE-MS anion analyses. Essentially complete ionization and increased hydrophobicity after the derivatization also enhanced MS detection sensitivity (e.g. formic acid was detected at 0.5,pg). Simultaneous derivatization of one sample using two structurally similar reagents, N -butyl-4-aminomethyl-pyridinum iodide (BAMP) and N -hexyl-4-aminomethyl-pyridinum iodide, provided additional information for recognizing a carboxylic acid in an unknown sample. Moreover, characteristic fragmentation acquired by online CE-MS/MS allowed for identification and categorization of carboxylic acids. Applying this method on rat urine, we found 59 ions matching the characteristic patterns of carboxylic acids. From these 59, 32 ions were positively identified and confirmed with standards. For comparative analysis, 24 standard carboxylic acids were derivatized by chemically identical but isotopically distinct BAMP and N -butyl-d9-4-aminomethyl-pyridinium iodide, and their derivatization limits and linearity ranges were determined. Comparative analysis was also performed on two individual urine samples derivatized with BAMP and N -butyl-d9-4-aminomethyl-pyridinium iodide. The metabolite profiling variation between these two samples was clearly visualized. [source] Microchip-based small, dense low-density lipoproteins assay for coronary heart disease risk assessmentELECTROPHORESIS, Issue 9 2008Hua Wang Abstract Small, dense low-density lipoprotein (sdLDL) has been accepted as an emerging cardiovascular risk factor, and there has been an increasing interest in analytical methods for sdLDL profiling for diagnosis. Serum sdLDL may be measured by different laboratory techniques, but all these methods are laborious, time-consuming, and costly. Recently, we have demonstrated that a low-temperature bonding of quartz microfluidic chips for serum lipoproteins analysis (Zhuang, G., Jin, Q., Liu, J., Cong, H. et al., Biomed. Microdevices 2006, 8, 255,261). In contrast to this previous study, we chose SDS as anionic surfactant to modify both lipoproteins and the channel surface to minimize lipoprotein adsorption and improve the resolution of lipoprotein separation. Two major LDL subclass patterns including large, buoyant LDL (lLDL), sdLDL, and high-density lipoprotein (HDL) were effectively separated with high reproducibility. RSD values of the migration time (min) and peak areas of standard LDL and HDL were 6.28, 4.02, 5.02, and 2.5%, respectively. Serum lipoproteins of 15 healthy subjects and 15 patients with coronary heart disease (CHD) were separated by microchip CE. No peaks of sdLDL were detected in serum samples of healthy subjects while sdLDL fractional peaks were observed in patients' entire serum samples. These results suggested that the microchip-based sdLDLs assay was a simple, rapid, and highly efficient technique and significantly improved the analysis of CHD risk factors. [source] Development of a daphnia magna DNA microarray for evaluating the toxicity of environmental chemicalsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 4 2007Hajime Watanabe Abstract Toxic chemical contaminants have a variety of detrimental effects on various species, and the impact of pollutants on ecosystems has become an urgent issue. However, the majority of studies regarding the effects of chemical contaminants have focused on vertebrates. Among aquatic organisms, Daphnia magna has been used extensively to evaluate organism- and populationlevel responses of invertebrates to pollutants in acute toxicity or reproductive toxicity tests. Although these types of tests can provide information concerning hazardous concentrations of chemicals, they provide no information about their mode of action. Recent advances in molecular genetic techniques have provided tools to better understand the responses of aquatic organisms to pollutants. In the present study, we adapted some of the techniques of molecular genetics to develop new tools, which form the basis for an ecotoxicogenomic assessment of D. magna. Based on a Daphnia expressed sequence tag database, we developed an oligonucleotide-based DNA microarray with high reproducibility. The DNA microarray was used to evaluate gene expression profiles of neonatal daphnids exposed to several different chemicals: Copper sulfate, hydrogen peroxide, pentachlorophenol, or ,-naphthoflavone. Exposure to these chemicals resulted in characteristic patterns of gene expression that were chemical-specific, indicating that the Daphnia DNA microarray can be used for classification of toxic chemicals and for development of a mechanistic understanding of chemical toxicity on a common freshwater organism. [source] Substance induced plasticity in noradrenergic innervation of the paraventricular hypothalamic nucleusEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 2 2003Arthur S. P. Jansen Abstract Single administration of the cytokine interleukin-1, (IL-1), or the psychostimulant amphetamine, enhanced adrenocorticotropin hormone and corticosterone responses to a stress challenge weeks later. This long-lasting hypothalamic-pituitary-adrenal (HPA)-sensitization is paralleled by an increase in electrically evoked release of noradrenaline in the paraventricular hypothalamic nucleus (PVN). We hypothesized that these functional changes may be associated with morphological plasticity of noradrenergic projections to the PVN, a parameter that shows high reproducibility. Specific alterations in relative (nor)adrenergic innervation density were studied by using dopamine-,-hydroxylase (DBH) as a marker. An image analysis system was used to detect changes in the relative DBH innervation density of the PVN. Groups of adult male rats were given IL-1 (10 µg/kg i.p.), amphetamine (5 mg/kg i.p.), or saline. Three weeks later, IL-1 and amphetamine primed rats showed enhanced adrenocorticotropin hormone and corticosterone responses to an amphetamine challenge. In another set of experiments, the relative DBH innervation density was measured in different PVN subnuclei at four rostro-caudal levels. Single administration of either IL-1 or amphetamine causes three weeks later a selective decrease in relative DBH innervation density in those subnuclei of the PVN that contain high numbers of corticotrophin-releasing hormone (CRH) producing neurons: the dorsal parvocellular and medial parvocellular PVN. We conclude that (1) long-lasting sensitization induced by single exposure to IL-1 and amphetamine induces specific pattern of neuroplastic changes in (nor)adrenergic innervation in the PVN and (2) reduction of relative DBH innervation density in CRH-rich areas is associated with paradoxical increase of electrically evoked release of (nor)adrenaline. [source] Comparability of transfer function resultsEUROPEAN TRANSACTIONS ON ELECTRICAL POWER, Issue 2 2006J. Christian Abstract The transfer function (TF) is a comparative method to monitor the mechanical condition of transformer windings. To perform on-site diagnosis a high reproducibility of TF results is required. Consequently, there are certain restrictions for the determination of the TF using time domain records. This paper describes the effect of different signal processing procedures on the results of TF calculations. Window functions, digital filters, length and treatment of pre-trigger samples are aspects of special interest for the application of the TF method. Additionally, condition parameters of the device under test affect the results of TF measurements. The correlation of transformer temperatures and TF characteristics has been investigated as well as the dielectric effect of the insulating oil and the position of the tap changer. Finally, the effect of different test set-up concepts is presented. Copyright © 2005 John Wiley & Sons, Ltd. [source] Reliability of orthostatic responses in healthy men aged between 65 and 75 yearsEXPERIMENTAL PHYSIOLOGY, Issue 4 2005Tim J. Gabbett The purpose of this study was to investigate the short-, medium- and long-term reproducibility of cardiovascular responses during 90° head-up tilt (HUT) in healthy older men. Twenty-eight healthy male subjects aged 69 (95% confidence intervals, 68,70) years participated in the study. Eight subjects underwent duplicate 90° HUT tests on consecutive days, while 20 subjects underwent four 90° HUT tests performed at baseline, and after 1 week, 1 month and 1 year. Following a 20-min supine resting period, each subject was rapidly tilted to the upright vertical position (90° HUT) and remained in that position for 15 min. Beat-by-beat recordings of mean (MAP), systolic (SBP) and diastolic (DBP) pressures were made via Finapres, while heart rate (HR) was monitored continuously from an electrocardiogram. No significant test,retest differences (P > 0.05) were observed for the changes in HR, MAP, SBP or DBP during 90° HUT. These measurements demonstrated high reproducibility (intraclass correlation coefficient, r= 0.91,0.99, P < 0.05). The supine resting and tilted HR, MAP, SBP and DBP over the 1-week, 1-month and 1-year period were not significantly different (P > 0.05) from baseline, and demonstrated high reproducibility (intraclass correlation coefficient, r= 0.82,0.98, P < 0.05). The results of this study demonstrate that in healthy older men, cardiovascular responses during orthostasis are highly reproducible, and this reproducibility is maintained over a 12-month period. These findings demonstrate that the 90° HUT test offers a reproducible method of monitoring longitudinal orthostatic responses in healthy older men. [source] Multifunctional Au-Coated TiO2 Nanotube Arrays as Recyclable SERS Substrates for Multifold Organic Pollutants DetectionADVANCED FUNCTIONAL MATERIALS, Issue 17 2010Xuanhua Li Abstract A multifunctional Au-coated TiO2 nanotube array is made via synthesis of a TiO2 nanotube array through a ZnO template, followed by deposition of Au particles onto the TiO2 surface using photocatalytic deposition and a hydrothermal method, respectively. Such arrays exhibit superior detection sensitivity with high reproducibility and stability. In addition, due to possessing stable catalytic properties, the arrays can clean themselves by photocatalytic degradation of target molecules adsorbed to the substrate under irradiation with UV light into inorganic small molecules using surface-enhanced Raman spectroscopy (SERS) detection, so that recycling can be achieved. Finally, by detection of Rhodamine 6G (R6G) dye, herbicide 4-chlorophenol (4-CP), persistent organic pollutant (POP) dichlorophenoxyacetic acid (2,4-D), and organophosphate pesticide methyl-parathion (MP), the unique recyclable properties indicate a new route in eliminating the single-use problem of traditional SERS substrates and show promising applications for detecting other organic pollutants. [source] High-Performance Photoresponsive Organic Nanotransistors with Single-Layer Graphenes as Two-Dimensional ElectrodesADVANCED FUNCTIONAL MATERIALS, Issue 17 2009Yang Cao Abstract Graphene behaves as a robust semimetal with the high electrical conductivity stemming from its high-quality tight two-dimensional crystallographic lattice. It is therefore a promising electrode material. Here, a general methodology for making stable photoresponsive field effect transistors, whose device geometries are comparable to traditional macroscopic semiconducting devices at the nanometer scale, using cut graphene sheets as 2D contacts is detailed. These contacts are produced through oxidative cutting of individual 2D planar graphene by electron beam lithography and oxygen plasma etching. Nanoscale organic transistors based on graphene contacts show high-performance FET behavior with bulk-like carrier mobility, high on/off current ratio, and high reproducibility. Due to the presence of photoactive molecules, the devices display reversible changes in current when they are exposed to visible light. The calculated responsivity of the devices is found to be as high as ,8.3,A,W,1. This study forms the basis for making new types of ultrasensitive molecular devices, thus initiating broad research interest in the field of nanoscale/molecular electronics. [source] Discovery of New Green Phosphors and Minimization of Experimental Inconsistency Using a Multi-Objective Genetic Algorithm-Assisted Combinatorial MethodADVANCED FUNCTIONAL MATERIALS, Issue 11 2009Asish Kumar Sharma Abstract A multi-objective genetic algorithm-assisted combinatorial materials search (MOGACMS) strategy was employed to develop a new green phosphor for use in a cold cathode fluorescent lamp (CCFL) for a back light unit (BLU) in liquid crystal display (LCD) applications. MOGACMS is a method for the systematic control of experimental inconsistency, which is one of the most troublesome and difficult problems in high-throughput combinatorial experiments. Experimental inconsistency is a very serious problem faced by all scientists in the field of combinatorial materials science. For this study, experimental inconsistency and material property were selected as dual objective functions that were simultaneously optimized. Specifically, in an attempt to search for promising phosphors with high reproducibility, luminance was maximized and experimental inconsistency was minimized using the MOGACMS strategy. A divalent manganese-doped alkali alkaline germanium oxide system was screened using MOGACMS. As a result of MOGA reiteration, we identified a phosphor, Na2MgGeO4:Mn2+, with improved luminance and reliable reproducibility. [source] Controlled sources for shear-wave surveys in minesGEOPHYSICAL PROSPECTING, Issue 3 2000Gordon M. Holmes The ability to analyse shear-wave anisotropy in a mine environment is greatly aided by using multiple source orientations of a reproducible, impulsive shear-wave source. The analysis of what is probably the first controlled source shear-wave experiment in a mine environment demonstrates clearly that shear-wave polarizations and time delays between split shear-wave arrivals are reliably measured because of the use of multiple source orientations rather than a single shear-wave source. Reliability is further aided by modelling the shear-wave source radiation pattern, which allows for the unequivocal discrimination between seismic raypaths where shear-wave splitting did and did not occur. The analysis also demonstrates the great importance of high reproducibility of the seismic source for the use of shear waves in time-lapse surveys to monitor changes in a rockmass. [source] An Improved Process for the N-Demethylation of Opiate Alkaloids using an Iron(II) Catalyst in Acetate BufferADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 1-2 2009Gaik Kok Abstract An improved process to N-demethylate opiate alkaloids utilising a solution of the ferrous porphyrin, tetrasodium 5,10,15,20-tetra(4-sulfophenyl)porphyrinatoiron(II) [=Fe(II)-TPPS (8)], in acetate buffer is described. This method provided the corresponding N-demethylated opiates in good yield with high reproducibility. [source] Imaging biomarkers of cardiovascular diseaseJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 3 2010Jinnan Wang PhD Abstract Cardiovascular disease (CVD) is a leading cause of morbidity and mortality worldwide. Current clinical techniques that rely on stenosis measurement alone appear to be insufficient for risk prediction in atherosclerosis patients. Many novel imaging methods have been developed to study atherosclerosis progression and to identify new features that can predict future clinical risk. MRI of atherosclerotic vessel walls is one such method. It has the ability to noninvasively evaluate multiple biomarkers of the disease such as luminal stenosis, plaque burden, tissue composition and plaque activity. In addition, the accuracy of in vivo MRI has been validated against histology with high reproducibility, thus paving the way for application to epidemiological studies of disease pathogenesis and, by serial MRI, in monitoring the efficacy of therapeutic intervention. In this review, we describe the various MR techniques used to evaluate aspects of plaque progression, discuss imaging-based measurements (imaging biomarkers), and also detail their validation. The application of plaque MRI in clinical trials as well as emerging imaging techniques used to evaluate plaque compositional features and biological activities are also discussed. J. Magn. Reson. Imaging 2010;32:502,515. © 2010 Wiley-Liss, Inc. [source] Quantitative analysis of MRI signal abnormalities of brain white matter with high reproducibility and accuracyJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 2 2002Xingchang Wei MD Abstract Purpose To assess the reproducibility and accuracy compared to radiologists of three automated segmentation pipelines for quantitative magnetic resonance imaging (MRI) measurement of brain white matter signal abnormalities (WMSA). Materials and Methods WMSA segmentation was performed on pairs of whole brain scans from 20 patients with multiple sclerosis (MS) and 10 older subjects who were positioned and imaged twice within 30 minutes. Radiologist outlines of WMSA on 20 sections from 16 patients were compared with the corresponding results of each segmentation method. Results The segmentation method combining expectation-maximization (EM) tissue segmentation, template-driven segmentation (TDS), and partial volume effect correction (PVEC) demonstrated the highest accuracy (the absolute value of the Z-score was 0.99 for both groups of subjects), as well as high interscan reproducibility (repeatability coefficient was 0.68 mL in MS patients and 1.49 mL in aging subjects). Conclusion The addition of TDS to the EM segmentation and PVEC algorithms significantly improved the accuracy of WMSA volume measurements, while also improving measurement reproducibility. J. Magn. Reson. Imaging 2002;15:203,209. © 2002 Wiley-Liss, Inc. [source] A new strategy based on recombinant viruses as a tool for assessing drug susceptibility of human immunodeficiency virus type 1JOURNAL OF MEDICAL VIROLOGY, Issue 2 2007J. Garcia-Perez Abstract The emergence of drug-resistant variants during antiretroviral therapy is a serious obstacle to sustained suppression of the human immunodeficiency virus type 1 (HIV-1). For that reason, resistance assays are essential to guide clinicians in the selection of optimal treatment regimens. Genotypic assays are less expensive and results are available faster than phenotypic assays. However, in heavily experienced patients with multiple treatment failures interpretation of complex mutation patterns remains difficult, and in these cases phenotypic assays are recommended. This report describes a novel recombinant virus assay where protease (PR) and reverse transcriptase (RT) sequences derived from the plasma isolated from patients are introduced into the back-bone of an HIV molecular clone that expresses Renilla luciferase protein in the place of nef gene. All drug resistance profiles analyzed correlate with previously reported data and showed high reproducibility. This assay, in addition to a fast (completed in 10 days), precise, reproducible and automated method, presents several advantages as compared to other phenotypic assays. The system described below allows the generation of recombinant viruses with multiples cycles of replication carrying a reporter gene in their genomes. These features increase the sensitivity of the test, an important aspect to be considered in the evaluation of less fit viral isolates. In conclusion, the assay permits the quantitation of the level of resistance of clinical HIV-1 isolates to PR and RT inhibitors. J. Med. Virol. 79:127,137, 2007. © 2006 Wiley-Liss, Inc. [source] Nanostructured gold surfaces as reproducible substrates for surface-enhanced Raman spectroscopyJOURNAL OF RAMAN SPECTROSCOPY, Issue 3 2007M. Sackmann Abstract Raman spectroscopy is a common tool for the qualitative and quantitative chemical analysis of molecules. Although the unique identification of molecules is possible via their vibrational lines, high concentrations (mmol/l) are needed for their nonresonant excitation owing to their low scattering cross section. The intensity of the Raman spectra is amplified by the use of the surface-enhanced Raman scattering (SERS) technique. While the use of silver sols results only in a limited reproducibility of the Raman line intensities, lithographically designed, nanostructured gold surfaces used as SERS-active substrates should, in principle, combine the high sensitivity with better reproducibility. For this purpose, we have produced gratings of gold dots on Si(001) surfaces by means of electron beam lithography. Qualitative and quantitative investigations of crystal violet (CV) performed using nanostructured surfaces give high reproducibility and enhancement of the Raman lines. The substrates are reusable after cleaning; all results presented could be obtained from a single SERS substrate. For the experiments very low laser powers were used. Copyright © 2006 John Wiley & Sons, Ltd. [source] Poly(N -vinylimidazole)-grafted capillary for electrophoresis prepared by surface-initiated atom transfer radical polymerizationJOURNAL OF SEPARATION SCIENCE, JSS, Issue 17-18 2010Jing Li Abstract A CE method for poly(N -vinylimidazole) (PVI)-grafted capillaries by the surface-initiated atom transfer radical polymerization has been developed. The coating was prepared with N -vinylimidazole as the monomer, 2-bromo-2-methyl- N -[3-(triethoxysilyl) propyl] propanamide (BTPAm) as the initiator and CuCl/CuCl2/2,2,-bipyridine as the catalyst and ligand. The direction and magnitude of EOF in the PVI-grafted capillary were investigated in a pH range of 3.0,9.0. The results indicated that the EOF could be modulated by varying the pH value of the buffer and an anodic EOF was obtained at pH values below 6.5. A significant improvement in reproducibility and reduction of EOF appeared on the PVI-grafted capillary when compared with the uncoated capillary. Furthermore, the polymer coated capillaries were applied to the separations of the inorganic anions, organic acids and basic proteins and baseline separations were achieved with short analysis time and high reproducibility. [source] Quantitative capillary electrophoresis and its application in analysis of alkaloids in tea, coffee, coca cola, and theophylline tabletsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 2 2009Mengjia Li Abstract A quantitative CE (qCE) system with high precision has been developed, in which a 4-port nano-valve was isolated from the electric field and served as sample injector. The accurate amount of sample was introduced into the CE system with high reproducibility. Based on this system, consecutive injections and separations were performed without voltage interruption. Reproducibilities in terms of RSD lower than 0.8% for retention time and 1.7% for peak area were achieved. The effectiveness of the system was demonstrated by the quantitative analysis of caffeine, theobromine, and theophylline in real samples, such as tea leaf, roasted coffee, coca cola, and theophylline tablets. [source] Analysis of an antibody pharmaceutical, tocilizumab, by capillary electrophoresis using a carboxylated capillaryJOURNAL OF SEPARATION SCIENCE, JSS, Issue 5 2008Atsushi Taga Abstract Antibody pharmaceuticals are becoming more and more prevalent due to their excellent effectiveness in clinical medications, and are expected to allow tailor-made medical treatment for rheumatic diseases, immunosuppression in cardiac transplantation, and cancer. Antibody-type pharmaceuticals of immunoglobulin G (IgG) commonly have N -glycosylated carbohydrate chains attached to heavy chains. The carbohydrate chains play important roles in the effectiveness of antibodies. Therefore evaluation of a glycosylated species is important in the first step of quality control of antibody pharmaceuticals. In the present work, we examined capillary electrophoresis with a newly developed, chemically modified capillary, the inner surface of which is modified with carboxyl groups, for evaluation of IgG molecular species which have carbohydrate chains; tocilizumab was used as a model. The analytical system developed in the present study is useful for determining the content of non-glycosylated peptides. In the analysis of tocilizumab, the ratio of non-glycosylated peptide was estimated to be 1.23% with a relative standard deviation of 3.05%. The method affords high reproducibility with simple operation, and analysis can be completed within 6 min. [source] Hydrothermal Synthesis of Alpha Alumina (,-Al2O3) Powders: Study of the Processing Variables and Growth MechanismsJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 2 2010Wojciech L. Suchanek Alpha alumina (,-Al2O3) powders and ,-Al2O3/boehmite (,-AlOOH) mixtures with controlled ,-AlOOH contents were synthesized hydrothermally under alkaline or acidic conditions at 380°,435°C for 1,10 days, under 6.9,14.5 MPa pressure, from concentrated precursors without stirring. The precursors were formed by mixing different types of aluminum hydroxides with water, and optionally with ,-Al2O3 seeds, hydrogen peroxide, sulfuric acid, dopants (i.e., KMnO4), and/or other additives. The experiments were performed on industrial scale in large production autoclaves. The synthesized ,-Al2O3 powders exhibited up to 100% phase purity, 99.98% chemical purity, equiaxed morphology, low aggregation levels, narrow crystallite size distributions with primary particle sizes ranging between 100 nm and 40 ,m, and high reproducibility. Precursor types, seeds, chemical additives, and temperature/time of the hydrothermal synthesis were found to govern properties of the powders. Different growth mechanisms for nanosized and rough powders are discussed. Results of this study enable the use of hydrothermal ,-Al2O3 powders in a multitude of applications, and make their hydrothermal production a commercial reality. [source] Dye-Sensitized Solar Cells Based on TiO2 Coatings with Dual Size-Scale PorosityJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 9 2009Lai Qi Dye-sensitized solar cells (DSSC) with efficiencies greater than 4% were produced with templated "inverse opal" titania coatings. A novel one-step method produces uniform and crack-free coatings made using commercially available titania nanoparticles with high reproducibility and uniformity. In this research, a volatile solvent electrolyte was tested; however, it shows proof-of-concept that larger pore volumes can be created for increased penetration of more viscous electrolytes that can be utilized in high-efficiency cells. This dual size-scale porosity film is a promising structure for DSSC applications, especially for those solid-state or quasi-solid-state cells that require polymer electrolytes. [source] Molecular typing of Aspergillus speciesMYCOSES, Issue 6 2008H. A. De Valk Summary Aspergillus species are widely distributed fungi that release large amounts of airborne conidia, which are dispersed in the environment. Several Aspergillus species have been described as human pathogens. Molecular techniques have been developed to investigate the epidemiological relation between environmental and clinical isolates. Several typing methods have been described for Aspergillus species, most of them with reference to Aspergillus fumigatus. Here, we summarise all the different available molecular typing techniques for Aspergillus. The performance of these techniques is evaluated with respect to their practical feasibility, and their interpretation and discriminatory power assessed. For A. fumigatus isolates, a large extent of genetic variability is demonstrated and therefore fingerprinting techniques with high discriminatory power and high reproducibility are required for this species. Afut1-restriction fragment length polymorphism and microsatellite typing showed the highest discriminatory power. In addition, the microsatellites show excellent reproducibility. Other typing techniques are still useful for smaller epidemiological problems and for less well-equipped laboratories. [source] Highly stable trypsin-aggregate coatings on polymer nanofibers for repeated protein digestionPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 7 2009Byoung Chan Kim Abstract A stable and robust trypsin-based biocatalytic system was developed and demonstrated for proteomic applications. The system utilizes polymer nanofibers coated with trypsin aggregates for immobilized protease digestions. After covalently attaching an initial layer of trypsin to the polymer nanofibers, highly concentrated trypsin molecules are crosslinked to the layered trypsin by way of a glutaraldehyde treatment. This process produced a 300-fold increase in trypsin activity compared with a conventional method for covalent trypsin immobilization, and proved to be robust in that it still maintained a high level of activity after a year of repeated recycling. This highly stable form of immobilized trypsin was resistant to autolysis, enabling repeated digestions of BSA over 40,days and successful peptide identification by LC-MS/MS. This active and stable form of immobilized trypsin was successfully employed in the digestion of yeast proteome extract with high reproducibility and within shorter time than conventional protein digestion using solution phase trypsin. Finally, the immobilized trypsin was resistant to proteolysis when exposed to other enzymes (i.e., chymotrypsin), which makes it suitable for use in "real-world" proteomic applications. Overall, the biocatalytic nanofibers with trypsin aggregate coatings proved to be an effective approach for repeated and automated protein digestion in proteomic analyses. [source] Proteome analysis of Schizosaccharomyces pombe by two-dimensional gel electrophoresis and mass spectrometryPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 14 2006Kyung-Hoon Hwang Abstract The fission yeast Schizosaccharomyces pombe (S. pombe) is a unicellular eukaryote and contains many genes and regulatory mechanisms that are close to those of mammals. In this study, we performed a global proteomic analysis of the fission yeast S. pombe wild type h,S L 972 proteome. More than 1500 protein spots were visualized on silver stained 2-D gels in the 3,10 pI range with a high resolution and high reproducibility. Protein identification was carried out by MALDI-TOF-MS and/or nanoLC-MS/MS. Advantage of the complementarity of these two MS approaches was used to enhance the identification quality. So far, 364 proteins (representing 157 different proteins) have been identified. We report here the identification of 117 new proteins on our 2-D reference map of this yeast compared to the first reference map. Of these identified proteins, 40.1% were involved in metabolism. The present work provides a very useful tool for all studies relying on S. pombe as a model organism and is a considerable complement to the first reference map of S. pombe published recently by Sun and coworkers (Sun, N., Jang, J., Lee, S., Kim, S. et al.., Proteomics 2005, 5, 1574,1579). [source] Visualization of protein interactions in living plant cells using bimolecular fluorescence complementationTHE PLANT JOURNAL, Issue 3 2004Michael Walter Summary Dynamic networks of protein,protein interactions regulate numerous cellular processes and determine the ability to respond appropriately to environmental stimuli. However, the investigation of protein complex formation in living plant cells by methods such as fluorescence resonance energy transfer has remained experimentally difficult, time consuming and requires sophisticated technical equipment. Here, we report the implementation of a bimolecular fluorescence complementation (BiFC) technique for visualization of protein,protein interactions in plant cells. This approach relies on the formation of a fluorescent complex by two non-fluorescent fragments of the yellow fluorescent protein brought together by association of interacting proteins fused to these fragments (Hu et al., 2002). To enable BiFC analyses in plant cells, we generated different complementary sets of expression vectors, which enable protein interaction studies in transiently or stably transformed cells. These vectors were used to investigate and visualize homodimerization of the basic leucine zipper (bZIP) transcription factor bZIP63 and the zinc finger protein lesion simulating disease 1 (LSD1) from Arabidopsis as well as the dimer formation of the tobacco 14-3-3 protein T14-3c. The interaction analyses of these model proteins established the feasibility of BiFC analyses for efficient visualization of structurally distinct proteins in different cellular compartments. Our investigations revealed a remarkable signal fluorescence intensity of interacting protein complexes as well as a high reproducibility and technical simplicity of the method in different plant systems. Consequently, the BiFC approach should significantly facilitate the visualization of the subcellular sites of protein interactions under conditions that closely reflect the normal physiological environment. [source] Secondary effects induced by the colon carcinogen azoxymethane in BDIX rats,APMIS, Issue 6 2004MORTEN KOBÆK-LARSEN Azoxymethane (AOM) is claimed to be a colon-specific carcinogen. In our studies, AOM was administered to adult BDIX/OrlIco rats by four weekly subcutaneous injections of 15 mg/kg body weight each , two periods of 2 weeks of AOM treatment separated by a one-week break. This treatment schedule resulted in colon carcinomas with a high frequency (75,100%) and with a high reproducibility. However, some serious side effects are associated with this carcinogen treatment. In addition to the colorectal tumours, we found small intestinal tumours, hepatic lesions and a high frequency of mesenchymal renal tumours which increased with longer latency periods. The renal tumours were only found in female rats, and this indicates a possible relation to sex hormones. We therefore analyzed both male and female kidneys for the expression of estrogen and progesterone receptors by immunohistochemical methods. A positive nuclear reaction for estrogen receptor was present in most tumour cells in all tumours and occasionally in nuclei of entrapped tubular cells, but never in glomeruli. Normal appearing renal tissue from female rats showed no positive reaction, but in male rats a slight nuclear reaction was seen in tubuli in the peripheral part of the medulla. A similar pattern was seen for progesterone receptors, but less pronounced. No rats developed tumours in the external ear canal, which is in contrast to studies performed in other rat strains. This may therefore be strain related. In order to reduce the secondary effects of the induction of colon cancer by AOM, it is advisable to use male rats only and a maximum latency period of 32 weeks. 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