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Reproducible Manner (reproducible + manner)
Selected AbstractsPreparation of Nanogapped Gold Nanoparticle Array for DNA DetectionELECTROANALYSIS, Issue 4 2008Shiho Tokonami Abstract A novel DNA detection technique using a gold nanoparticle array film electrode has been reported here. The gold nanoparticles molecularly linked with binder molecule (1,10-decanedithiol) were separated 1.3,nm from each other, and the DNA conductivity change from single to double strand was measured by monitoring a voltage drop across the particles, between which a probe of a 12-mer oligonucleotide was immobilized. In adding a complementary oligonucleotide on the nanoparticle film chip, an immediate decrease in the film resistance (ca. 1.4 ,) due to a hybridization event occurred in a reproducible manner with this simple setup. In the paper, we have an interest in the primary sensing properties; effect of the film resistance on the sensor response, dependence of the resistance change on the DNA concentration, and the performance of the system for DNA detection including single nucleotide polymorphisms were described. [source] High-Density Carrier Accumulation in ZnO Field-Effect Transistors Gated by Electric Double Layers of Ionic LiquidsADVANCED FUNCTIONAL MATERIALS, Issue 7 2009Hongtao Yuan Abstract Very recently, electric-field-induced superconductivity in an insulator was realized by tuning charge carrier to a high density level (1,×,1014 cm,2). To increase the maximum attainable carrier density for electrostatic tuning of electronic states in semiconductor field-effect transistors is a hot issue but a big challenge. Here, ultrahigh density carrier accumulation is reported, in particular at low temperature, in a ZnO field-effect transistor gated by electric double layers of ionic liquid (IL). This transistor, called an electric double layer transistor (EDLT), is found to exhibit very high transconductance and an ultrahigh carrier density in a fast, reversible, and reproducible manner. The room temperature capacitance of EDLTs is found to be as large as 34,µF cm,2, deduced from Hall-effect measurements, and is mainly responsible for the carrier density modulation in a very wide range. Importantly, the IL dielectric, with a supercooling property, is found to have charge-accumulation capability even at low temperatures, reaching an ultrahigh carrier density of 8×1014 cm,2 at 220,K and maintaining a density of 5.5×1014 cm,2 at 1.8,K. This high carrier density of EDLTs is of great importance not only in practical device applications but also in fundamental research; for example, in the search for novel electronic phenomena, such as superconductivity, in oxide systems. [source] The innervation of FGF-induced additional limbs in the chick embryoJOURNAL OF ANATOMY, Issue 1 2003B. W. Turney Abstract Motoneurones that supply the vertebrate limb innervate their muscle targets in a highly reproducible manner. As development proceeds, these limb-specific motoneurones send out axons, which grow towards the developing limb and then congregate at its base to form the plexus. In the plexus, in response to unknown positional cues, these axons rearrange, often changing their original spatial relationships, before sorting out to emerge in the defined nerve trunks that innervate the limb. Several proposals have been put forward to explain how this reproducible innervation pattern is achieved. These include (1) that early differences in the motoneurone identity dictate their future axonal trajectories, (2) that axons actively respond to attractive or repulsive positional cues provided by the limb bud itself, or (3) that motor axons are passively deployed, following pathways of least mechanical resistance. We have addressed the question of the relative roles of motoneurone identity and the signals that the axons encounter on their journey towards the limb bud. Using the developing chick embryo as our experimental model we tested the effect of providing an additional limb target for motor axons leaving the flank level of the spinal cord. To do this we placed FGF-soaked beads in the presumptive flank of 2-day-old chick embryos. This treatment induces an additional limb containing muscles. We investigated whether such additional limbs are innervated and by which neurones. We show that rather than the additional limbs being solely supplied by axons diverted from the two existing limb plexuses, motoneurones that normally supply the flank alter their trajectories to enter the induced limb. Once in the limb, axons respond to positional cues within the bud to generate the stereotypical innervation pattern. Our results show that the tendency of ,flank' motoneurones to innervate flank can be overcome by the presence of an additional limb. [source] Assessing normal pulse wave velocity in the proximal pulmonary arteries using transit time: A feasibility, repeatability, and observer reproducibility study by cardiovascular magnetic resonanceJOURNAL OF MAGNETIC RESONANCE IMAGING, Issue 5 2007MRCP, William M. Bradlow BM Abstract Purpose To calculate pulse wave velocity (PWV) in the proximal pulmonary arteries (PAs) by cardiovascular magnetic resonance (CMR) using the transit-time method, and address respiratory variation, repeatability, and observer reproducibility. Materials and Methods A 1.9-msec interleaved phase velocity sequence was repeated three times consecutively in 10 normal subjects. Pulse wave (PW) arrival times (ATs) were determined for the main and branch PAs. The PWV was calculated by dividing the path length traveled by the difference in ATs. Respiratory variation was considered by comparing acquisitions with and without respiratory gating. Results For navigated data the mean PWVs for the left PA (LPA) and right PA (RPA) were 2.09 ± 0.64 m/second and 2.33 ± 0.44 m/second, respectively. For non-navigated data the mean PWVs for the LPA and RPA were 2.14 ± 0.41 m/second and 2.31 ± 0.49 m/second, respectively. No statistically significant difference was found between respiratory non-navigated data and navigated data. Repeated on-table measurements were consistent (LPA non-navigated P = 0.95, RPA non-navigated P = 0.91, LPA navigated P = 0.96, RPA navigated P = 0.51). The coefficients of variation (CVs) were 12.2% and 12.5% for intra- and interobserver assessments, respectively. Conclusion One can measure PWV in the proximal PAs using transit-time in a reproducible manner without respiratory gating. J. Magn. Reson. Imaging 2007;25:974,981. © 2007 Wiley-Liss, Inc. [source] Proposed model of botulinum toxin-induced muscle weakness in the rabbitJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 6 2005D. Longino Abstract Osteoarthritic patients show only a weak association between radiographic signs of joint disease and joint pain and disability. Conversely, muscle weakness is one of the earliest and most common symptoms of patients with osteoarthritis (OA). However, while many experimental models of osteoarthritis include a component of muscular weakness, no model has isolated this factor satisfactorily. Therefore, the purpose of this study was to develop and validate an experimental animal model of muscle weakness for future use in the study of OA. Botulinum Type-A toxin (BTX-A) was uni-laterally injected into the quadriceps musculature of New Zealand white rabbits (3.5 unit/kg). Isometric knee extensor torque at a range of knee angles and stimulation frequencies, and quadriceps muscle mass, were quantified for control animals, and at one- and six-months post-repeated injections, in both, the experimental and the contralateral hindlimb. Ground reaction forces were measured in all animals while hopping across two force platforms. Isometric knee extension torque and quadriceps muscle mass was systematically decreased in the experimental hindlimb. Vertical ground reaction forces in the push off phase of hopping were also decreased in the experimental compared to control hindlimbs. We conclude that BTX-A injection into the rabbit musculature creates functional and absolute muscle weakness in a reproducible manner. Therefore, this model may be used to systematically study the possible effects of muscle weakness on joint degeneration, either as an isolated intervention, or in combination with other interventions (anterior cruciate ligament transection, meniscectomy) known to create knee joint degeneration. © 2005 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved. [source] Inhibition of serum angiotensin-converting enzyme in rabbits after intravenous administration of enalaprilat-loaded intact erythrocytesJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 9 2001Mehrdad Hamidi Encapsulation of drugs in intact erythrocytes, because of the profound characteristics of these natural microspheres, has gained considerable attention in recent years. In this study, the inhibition time courses of serum angiotensin-converting enzyme (ACE) activity after intravenous administration of enalaprilat encapsulated in intact erythrocytes was evaluated and compared with free drug, in a rabbit model. Three groups of animals each received free drug, drug-loaded erythrocytes or sham-encapsulated erythrocytes. Serum ACE activity was determined in each case using the synthetic substrate hippuryl-histidyl-leucine and quantitation of the hippuric acid released by a developed and validated HPLC method. The serum ACE inhibition profiles in the three groups showed that the encapsulated drug inhibited the serum ACE more slowly, more efficiently, over a considerably longer time and in a more reproducible manner, than the free drug or sham-encapsulated erythrocytes. We conclude that the erythrocytes can serve as efficacious slow-release drug carriers for enalaprilat in circulation. [source] Reliability of the Amsterdam Clinical Challenge Scale (ACCS): a new instrument to assess the level of difficulty of patient cases in medical educationMEDICAL EDUCATION, Issue 7 2000Gercama Introduction In problem-based medical curricula, consideration should be given to the level of difficulty of patient cases used for training and assessment. The Amsterdam Clinical Challenge Scale (ACCS) has been developed to assess the degree of difficulty of patient cases in a systematic and reproducible manner. To determine the reliability of the instrument two research questions were addressed: (1) How many judges are required, on the basis of the total score of the ACCS, to obtain a reliable estimate of the difficulty of a single case? (2) How many cases and/or how many judges are needed to reach an acceptable level of reliability of the total score of the ACCS? Method Four judges scored 36 patient scripts reflecting a wide range of patient problems encountered in general practice. Each script was scored four times. In the reliability analysis, the generalizability theory was applied. Results The results show that the judges did, indeed, use the whole range of difficulty ratings. When the ACCS is applied to a single case, eight or more judges are needed to reach an acceptable level of reliability. When more cases are involved, fewer judges are needed; for 10 or more cases one judge will be sufficient. Conclusions Given the typical length, for example of an objective structured clinical examination, the ACCS makes it possible to provide a reliable estimate of the level of difficulty of such a test with only a limited number of judges. [source] Computer-Based Analysis of Dynamic QT Changes: Toward High Precision and Individual Rate CorrectionANNALS OF NONINVASIVE ELECTROCARDIOLOGY, Issue 4 2002Corina Dota M.D. Background: New strategies are needed to improve the results of automatic measurement of the various parts of the ECG signal and their dynamic changes. Methods: The EClysis software processes digitally-recorded ECGs from up to 12 leads at 500 Hz, using strictly defined algorithms to detect the PQRSTU points and to measure ECG intervals and amplitudes. Calculations are made on the averaged curve of each sampling period (beat group) or as means ± SD for beat groups, after being analyzed at the individual beat level in each lead. Resulting data sets can be exported for further statistical analyses. Using QT and R-R measured on beat level, an individual correction for the R-R dependence can be performed. Results: EClysis assigns PQRSTU points and intervals in a sensitive and highly reproducible manner, with coefficients of variation in ECG intervals corresponding to ca. 2 ms in the simulated ECG. In the normal ECG, the CVs are 2% for QRS, 0.8% for QT, and almost 6% for PQ intervals. EClysis highlights the increase in QT intervals and the decrease of T-wave amplitudes during almokalant infusion versus placebo. Using the observed linear or exponential relationships to adjust QT for R-R dependence in healthy subjects, one can eliminate this dependence almost completely by individualized correction. Conclusions: The EClysis system provides a precise and reproducible method to analyze ECGs. A.N.E. 2002;7(4):289,301 [source] Generation of human embryonic stem cell-derived mesoderm and cardiac cells using size-specified aggregates in an oxygen-controlled bioreactorBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2009Sylvia Niebruegge Abstract The ability to generate human pluripotent stem cell-derived cell types at sufficiently high numbers and in a reproducible manner is fundamental for clinical and biopharmaceutical applications. Current experimental methods for the differentiation of pluripotent cells such as human embryonic stem cells (hESC) rely on the generation of heterogeneous aggregates of cells, also called "embryoid bodies" (EBs), in small scale static culture. These protocols are typically (1) not scalable, (2) result in a wide range of EB sizes and (3) expose cells to fluctuations in physicochemical parameters. With the goal of establishing a robust bioprocess we first screened different scalable suspension systems for their ability to support the growth and differentiation of hESCs. Next homogeneity of initial cell aggregates was improved by employing a micro-printing strategy to generate large numbers of size-specified hESC aggregates. Finally, these technologies were integrated into a fully controlled bioreactor system and the impact of oxygen concentration was investigated. Our results demonstrate the beneficial effects of stirred bioreactor culture, aggregate size-control and hypoxia (4% oxygen tension) on both cell growth and cell differentiation towards cardiomyocytes. QRT-PCR data for markers such as Brachyury, LIM domain homeobox gene Isl-1, Troponin T and Myosin Light Chain 2v, as well as immunohistochemistry and functional analysis by response to chronotropic agents, documented the impact of these parameters on cardiac differentiation. This study provides an important foundation towards the robust generation of clinically relevant numbers of hESC derived cells. Biotechnol. Bioeng. 2009;102: 493,507. © 2008 Wiley Periodicals, Inc. [source] Towards a Methodology for Developing Evidence-Informed Management Knowledge by Means of Systematic ReviewBRITISH JOURNAL OF MANAGEMENT, Issue 3 2003David Tranfield Undertaking a review of the literature is an important part of any research project. The researcher both maps and assesses the relevant intellectual territory in order to specify a research question which will further develop the knowledge base. However, traditional ,narrative' reviews frequently lack thoroughness, and in many cases are not undertaken as genuine pieces of investigatory science. Consequently they can lack a means for making sense of what the collection of studies is saying. These reviews can be biased by the researcher and often lack rigour. Furthermore, the use of reviews of the available evidence to provide insights and guidance for intervention into operational needs of practitioners and policymakers has largely been of secondary importance. For practitioners, making sense of a mass of often-contradictory evidence has become progressively harder. The quality of evidence underpinning decision-making and action has been questioned, for inadequate or incomplete evidence seriously impedes policy formulation and implementation. In exploring ways in which evidence-informed management reviews might be achieved, the authors evaluate the process of systematic review used in the medical sciences. Over the last fifteen years, medical science has attempted to improve the review process by synthesizing research in a systematic, transparent, and reproducible manner with the twin aims of enhancing the knowledge base and informing policymaking and practice. This paper evaluates the extent to which the process of systematic review can be applied to the management field in order to produce a reliable knowledge stock and enhanced practice by developing context-sensitive research. The paper highlights the challenges in developing an appropriate methodology. [source] Electrophysiological characterization of the SK channel blockers methyl-laudanosine and methyl-noscapine in cell lines and rat brain slicesBRITISH JOURNAL OF PHARMACOLOGY, Issue 6 2004Jacqueline Scuvée-Moreau We have recently shown that the alkaloid methyl-laudanosine blocks SK channel-mediated afterhyperpolarizations (AHPs) in midbrain dopaminergic neurones. However, the relative potency of the compound on the SK channel subtypes and its ability to block AHPs of other neurones were unknown. Using whole-cell patch-clamp experiments in transfected cell lines, we found that the compound blocks SK1, SK2 and SK3 currents with equal potency: its mean IC50s were 1.2, 0.8 and 1.8 ,M, respectively. IK currents were unaffected. In rat brain slices, methyl-laudanosine blocked apamin-sensitive AHPs in serotonergic neurones of the dorsal raphe and noradrenergic neurones of the locus coeruleus with IC50s of 21 and 19 ,M, as compared to 15 ,M in dopaminergic neurones. However, at 100 ,M, methyl-laudanosine elicited a constant hyperpolarization of serotonergic neurones of about 9 mV, which was inconsistently (i.e. not in a reproducible manner) antagonized by atropine and hence partly due to the activation of muscarinic receptors. While exploring the pharmacology of related compounds, we found that methyl-noscapine also blocked SK channels. In cell lines, methyl-noscapine blocked SK1, SK2 and SK3 currents with mean IC50s of 5.9, 5.6 and 3.9 ,M, respectively. It also did not block IK currents. Methyl-noscapine was slightly less potent than methyl-laudanosine in blocking AHPs in brain slices, its IC50s being 42, 37 and 29 ,M in dopaminergic, serotonergic and noradrenergic neurones, respectively. Interestingly, no significant non-SK effects were observed with methyl-noscapine in slices. At a concentration of 300 ,M, methyl-noscapine elicited the same changes in excitability in the three neuronal types than did a supramaximal concentration of apamin (300 nM). Methyl-laudanosine and methyl-noscapine produced a rapidly reversible blockade of SK channels as compared with apamin. The difference between the IC50s of apamin (0.45 nM) and methyl-laudanosine (1.8 ,M) in SK3 cells was essentially due to a major difference in their k,1 (0.028 s,1 for apamin and 20 s,1 for methyl-laudanosine). These experiments demonstrate that both methyl-laudanosine and methyl-noscapine are medium potency, quickly dissociating, SK channel blockers with a similar potency on the three SK subtypes. Methyl-noscapine may be superior in terms of specificity for the SK channels. British Journal of Pharmacology (2004) 143, 753,764. doi:10.1038/sj.bjp.0705979 [source] |