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At least 883 records · Page 49Linked to original sources
PubMed2000

Chromatographic assay of pharmaceutical compounds under column overloading.

A HPLC assay method utilizing overloaded chromatography and dual-wavelength detection was developed for a pharmaceutical formulation containing an antibacterial (clotrimazole) and a steroid (mometasone furoate) at widely different concentrations. In order to meet the limit of quantitation (LOQ) objective of not less than 0.05% of assay concentration simultaneously for both actives in the HPLC assay method, the assay concentration of the antibacterial falls into the non-linear range of its equilibrium isotherm, but still in the linear dynamic range of an ultraviolet detector. Although the analytical column is overloaded with the antibacterial and a non-symmetric elution peak is obtained, the HPLC assay method exhibits good linearity, recovery and reproducibility.

Chromatography, High Pressure Liquid↗
PubMed2001

The short-time Fourier transform and muscle fatigue assessment in dynamic contractions.

The mean frequency of the power spectrum of an electromyographic signal is an accepted index for monitoring fatigue in static contractions. There is however, indication that it may be a useful index even in dynamic contractions in which muscle length and/or force may vary. The objective of this investigation was to explore this possibility. An examination of the effects of amplitude modulation on modeled electromyographic signals revealed that changes in variance created in this way do not sufficiently affect characteristic frequency data to obscure a trend with fatigue. This validated the contention that not all non-stationarities in signals necessarily manifest in power spectral parameters. While an investigation of the nature and effects of non-stationarities in real electromyographic signals produced from dynamic contractions indicated that a more complex model is warranted, the results also indicated that averaging associated with estimating spectral parameters with the short-time Fourier transform can control the effects of the more complex non-stationarities. Finally, a fatigue test involving dynamic contractions at a force level under 30% of peak voluntary dynamic range, validated that it was possible to track fatigue in dynamic contractions using a traditional short-time Fourier transform methodology.

Adult↗
PubMed1995

Evaluation of charge-injection devices for use in laue diffraction imaging.

Charge-injection devices (CIDs) are versatile detectors having a number of features which recommend them for use in the imaging of X-ray diffraction patterns. They have a flexible nondestructive readout allowing for analysis of image quality during data collection and rapid readout of selected portions of the device. CIDs have full-well capacities in the range of 10(6) charge carriers giving them a high dynamic range for both direct and indirect imaging of X-rays. CIDs have peak quantum efficiencies in the optical region over 50% allowing for their incorporation into indirect detection systems. Rapid random single-pixel address allows for their use as single X-ray photon counters with energy discrimination. Three types of position-sensitive detectors for X-rays have been developed using CIDs. Two CID formats, the CID 17PPRA (388 x 256) and the CID 38SG (512 x 512), were incorporated into systems performing indirect imaging, direct imaging and single X-ray photon counting with energy discrimination. Indirect images of the Laue diffraction patterns from tetraphenylphosphonium tetrachlorooxomolybdenum(V) and natural MoS(2) were collected using a phosphor sheet to convert X-rays into optical photons which were detected with the CID 38SG. Directly detected images of spots from the Laue diffraction pattern of MoS(2) were recorded with the CID 17PPRA. Single photon counting with energy discrimination is demonstrated with the CID 17PPRA using a reflection from the Laue diffraction pattern of MoS(2). Useful information could be obtained from a single pixel at read rates over 7 kHz. Complete energy-dispersive analysis suitable for determination of space groups from Laue diffraction is currently limited due to incomplete charge collection and/or split events.

Journal Article↗
PubMed2001

The behavior of a poly(aniline) solid contact pH selective electrode based on N,N,N',N'-tetrabenzylethanediamine ionophore.

A hydrogen ion-selective solid-contact electrode based on N,N,N',N'-tetrabenzylethanediamine has shown the best Nernstian slope and selectivity and the widest response range in a Tris buffered pH sample solution. Its linear dynamic range was pH 3.50-11.94, and the Nernstian slope showed 52.1 mV/pH (at 20 +/- 0.2 degrees C). When it was directly applied to human whole blood (in pH range 6.0-8.5) we could obtain the same satisfying results. This electrode continuously contacted a Tris 7.47 buffered solution, human whole blood and a hydrofluoric acid solution for one month without any loss of performance. Also, hydrofluoric acid did not influence the surface of this electrode, and thus it was maintained without showing any changes in potentials after being used in a hydrofluoric acid solution. The standard deviation in the determined e.m.f. differences was 1.5 mV (N = 5) for Tris buffer solutions of pH 6.5 and 1.1 mV at a Tris buffer solutions of pH 8.5. The 90% response time of the electrodes obtained by injecting of hydrochloric acid into the Tris buffer sample solution was less than 10 s. Especially, in the this paper, with these potential response characteristics of hydrogen ion selective poly(aniline) solid contact electrode, we have also presented the pH response mechanism of this electrode and the role of poly(aniline) and a doped anion in a poly(aniline) layer.

Journal Article↗
PubMed2005

[Study on the interaction of tetraiodofluorescein and bovine serum albumin by fluorimetry].

The binding reaction of tetraiodofluorescein with bovine serum albumin i n aqueous solution was studied by fluorimetry.The binding constant is 2.65 x 10(5) L x mol(-1), the equilibrium constant is 5.7 x 10(4) L x mol(-1), and the binding number is 0.86. The research results of binding model indicate that the hydrophobic force was the main binding force. The presence of BSA can increase the fluorescence intensity of TIF. And the deltaF(F-F0) is linear to the concentration of BSA in a certain range, so the first analytical application was proposed. The linear dynamic range (LDR) was 8 x 10(-7) - 9 x 10(-6) mol x L(-1), the limit of detection (LOD) was 9.88 x 10(-8) mol x L(-1), and the relative standard deviation (RSD) was 3.4%.

Algorithms↗
PubMed1983

Automated radioautographic grain counting. Correction for grain overlap.

An algorithm is described for the calculation of radioautographic cell grain count from measurements of total cell nuclear area and total grain area. This algorithm provides a statistical correction for grain overlap that is based on the solution to the occupancy problem in probability theory. This method permits the use of automated grain counting over a wide range of grain counts per cell, and extends the useful dynamic range of radioautographic grain counting to well over 200 grains/cell.

Autoradiography↗
PubMed1995

Measurement of growth hormone-binding protein in the rat by a ligand immunofunctional assay.

We have developed a ligand immunofunctional assay (LIFA) for quantifying the circulating functional GH-binding protein (GHBP) in the rat. This two-site solid-phase assay uses a capture monoclonal antibody (4.3) specific to the hydrophilic C-terminal segment of rat GHBP (rGHBP), saturation of binding with human GH, and a detection system of rabbit antihuman GH polyclonal antibody and peroxidase-conjugated antirabbit immunoglobulin G antibody. Results were compared with Scatchard estimates derived by immuno-precipitation with monoclonal antibody 4.3. This assay was used to determine the GHBP levels in male and female rats and to investigate the diurnal properties and dynamics of GH and GHBP interaction in 15-min blood sampling over a 6-h period. The dynamic range of the rLIFA was 0.15-20.0 nM recombinant rGHBP, with intraassay and interassay coefficients of variation of 10.5% (n = 20) and 12.9% (n = 12), respectively. Serum GHBP levels determined by the rLIFA and those derived from Scatchard estimates were strongly correlated (n = 8; beta = 0.55; r2 = 0.89; P = 0.0005). Male rats had lower GHBP levels (6.5 +/- 0.7 nM; mean +/- SE; n = 14) than female rats (35.4 +/- 2.7 nM; n = 15; P = 0.0001). In the diurnal study, male rats had higher GH peaks (312.5 +/- 121.6 ng/ml; n = 7) than female rats (96.5 +/- 15.4 ng/ml; n = 9; P < 0.0001). In contrast to the pulsatile secretion of GH, GHBP levels in both sexes remained stable and showed no relationship to secretory pulses of GH. However, the GH bursts significantly altered the distribution of the GH-GHBP complex in male rats. By saturation and mass analysis, the greater GH pulsatile secretion in male rats resulted in occupancy of GHBP from less than 5% at nadir to about 80% at secretory peaks, in contrast to the less than 5-15% range of GHBP occupancy in female rats. In male rats, greater than 80% of GH at secretory peaks existed in the free form, whereas in female rats, 16-23% of GH existed in the free form during pulsatile secretion. In summary, the rLIFA shows good correlation to Scatchard analysis using an identical antibody. We conclude that this assay provides a rapid, sensitive, and accurate measurement of the circulating functional GHBP in the rat, and that it facilitates the study of GH and GHBP dynamics under a range of physiological conditions.

Animals↗
PubMed1995

An amplified chemiluminescent assay for the detection of alkaline phosphatase.

An amplification assay for the measurement of alkaline phosphatase has been combined with a luminescent end point using the luminol-peroxidase system to produce the first enzyme-amplified chemiluminescent assay based on the principle of prosthetogenesis. This assay is both quantitative and extremely sensitive. When the assay was used to detect alkaline phosphatase in solution, the detection limit was 0.4 amol in a 5-min assay. The interassay variance ranged from 4 to 20% and 7 to 19% across the dynamic range of the assay for a chemiluminescent assay and an enhanced chemiluminescent assay, respectively, employing two different preparations of luminol.

Alkaline Phosphatase↗
PubMed1996

Disposable sensor for biochemical oxygen demand.

Disposable-type microbial sensors were prepared for the determination of biochemical oxygen demand (BOD). The yeast, Trichosporon cutaneum, was directly immobilized on the surface of miniature oxygen electrodes using an ultraviolet crosslinking resin (ENT-3400). The oxygen electrodes (15 mm x 2 mm x 0.4 mm) were made on silicon substrates using micromachining techniques. They were Clark-type two-electrode systems with-1021 mV applied to the working electrode. Typical response times of the BOD sensors were in the range of 7-20 min. At 20 degrees C, the sensors' dynamic range was from 0 to 18 mg/1 BOD when a glucose/glutamate BOD standard solution was used. The lower limit of detection was 0.2 mg/l BOD. This value was one order of magnitude lower than that of sensors previously reported. The sensors' operational lifetime of 3 days was satisfactory for a disposable type. The sensors' responses were reproducible to within 8% relative standard deviation. The BOD sensors' were applied to untreated and treated waste waters from industrial effluents and municipal sewage. BOD values determined using these sensors correlated well with those determined by the conventional 5-day BOD determination method.

Biosensing Techniques↗
PubMed2001

Chemokine receptor homo- or heterodimerization activates distinct signaling pathways.

Chemokine receptors of both the CC and CXC families have been demonstrated to undergo a ligand-mediated homodimerization process required for Ca2+ flux and chemotaxis. We show that, in the chemokine response, heterodimerization is also permitted between given receptor pairs, specifically between CCR2 and CCR5. This has functional consequences, as the CCR2 and CCR5 ligands monocyte chemotactic protein-1 (MCP-1) and RANTES (regulated upon activation, normal T cell-expressed and secreted) cooperate to trigger calcium responses at concentrations 10- to 100-fold lower than the threshold for either chemokine alone. Heterodimerization results in recruitment of each receptor-associated signaling complex, but also recruits dissimilar signaling path ways such as G(q/11) association, and delays activation of phosphatidyl inositol 3-kinase. The consequences are a pertussis toxin-resistant Ca2+ flux and trig gering of cell adhesion rather than chemotaxis. These results show the effect of heterodimer formation on increasing the sensitivity and dynamic range of the chemokine response, and may aid in understanding the dynamics of leukocytes at limiting chemokine concentrations in vivo.

Calcium Signaling↗
PubMed1995

Imaging plates for use with synchrotron radiation.

This review summarizes the principles and performance characteristics of the imaging-plate (IP) X-ray area detector, which is based on the photo-stimulable phosphor BaF(Br,I):Eu(2+), together with some of its applications at the Photon Factory. The photo-stimulable phosphor can temporarily store an X-ray image. The stored image is read out by measuring the intensity of luminescence, which is stimulated by an He-Ne laser beam scanning the phosphor surface. The IP has a spatial resolution of 170 mum (FWHM) with a pixel size of 100 x 100 mum and area sizes ranging from 127 x 127 to 201 x 400mm(2). The dynamic range is over l:10(5). The detective quantum efficiency, which is a function of exposure level, is more than 80% for 8-20 keV X-rays at medium exposure levels. The background noise level is equivalent to less than 3 X-ray photons/pixel of 8 keV. The precision in intensity measurement is 0.5-1% at best. These performance characteristics of the IP depend largely on the performance of the IP readout system. Two applications of the IP to time-resolved measurements are discussed: one is based on a cinema method which achieves a 0.3 s time resolution for 40 exposures of size 127 x 127 mm(2), and the other uses the IP as a linear detector of length 200 mm with a 23 mus time resolution for a time period of 46 ms, based on the streak-camera method.

Journal Article↗
PubMed2004

NMDA-receptor regulation of muscarinic-receptor stimulated inositol 1,4,5-trisphosphate production and protein kinase C activation in single cerebellar granule neurons.

Inositol 1,4,5-trisphosphate (InsP(3)) production in single cerebellar granule neurons (CGNs) grown in culture was measured using the PH domain of phospholipase C delta1 tagged with enhanced green fluorescent protein (eGFP-PH(PLCdelta1)). These measurements were correlated with changes in intracellular free Ca2+ determined by single cell imaging. In control CGNs, intracellular Ca2+ stores appeared replete. However, the refilling state of these stores appeared dependent on the fluorophore used to measure Ca2+-release. Thus, methacholine (MCH), acting via muscarinic acetylcholine-receptors (mAchRs), mobilised intracellular Ca2+ in cells loaded with fluo-3 and fura-4f, but not fura-2. Confocal measurements of single CGNs expressing eGFP-PH(PLCdelta1) demonstrated that MCH stimulated a robust peak increase in InsP(3), which was followed by a sustained plateau phase of InsP(3) production. In contrast, glutamate-induced InsP(3) signals were weak or not detectable. MCH-stimulated InsP(3) production was reduced by chelation of intracellular Ca2+ with BAPTA, and emptying of intracellular stores with thapsigargin, indicated a positive feedback effect of Ca2+ mobilisation onto PLC activity. In CGNs, NMDA- and KCl-mediated Ca2+-entry significantly enhanced MCH-induced InsP(3) production. Furthermore, mAchR-mediated PLC activation appeared sensitive to the full dynamic range of intracellular Ca2+ increases stimulated by 100 microm NMDA. This dynamic regulation was also observed at the level of PKC activation indicated by an enhanced translocation of eGFP-tagged myristoylated alanine-rich C kinase substrate (MARCKS) protein in cells stimulated with MCH. Thus, NMDA-mediated Ca2+ influx and PLC activation may represent a coincident-detection system whereby ionotropic and metabotropic signals combine to stimulate InsP(3) production and PKC-mediated phosphorylation events in CGNs.

Animals↗
PubMed2002

Amplification of signaling events in bacteria.

Bacteria respond to extremely shallow chemical gradients by modifying their motility in a process called chemotaxis. This chemotactic response is characterized by high sensitivity to small concentration differences, which extends over a large range of concentrations. This combination of high signal gain and large dynamic range results from both a memory of past events and the ability to amplify small differences in signal between the memory and the current environment. Dahlquist describes the signaling mechanism used by bacteria to regulate the flagellar motor and the places in this pathway where signal amplification may occur.

Cell Movement↗
PubMed2006

Intrinsic bursters increase the robustness of rhythm generation in an excitatory network.

The pre-Botzinger complex (pBC) is a vital subcircuit of the respiratory central pattern generator. Although the existence of neurons with pacemaker-like bursting properties in this network is not questioned, their role in network rhythmogenesis is unresolved. Modeling is ideally suited to address this debate because of the ease with which biophysical parameters of individual cells and network architecture can be manipulated. We modeled the parameter variability of experimental data from pBC bursting pacemaker and nonpacemaker neurons using a modified version of our previously developed pBC neuron and network models. To investigate the role of pacemakers in networkwide rhythmogenesis, we simulated networks of these neurons and varied the fraction of the population made up of pacemakers. For each number of pacemaker neurons, we varied the amount of tonic drive to the network and measured the frequency of synchronous networkwide bursting produced. Both excitatory networks with all-to-all coupling and sparsely connected networks were explored for several levels of synaptic coupling strength. Networks containing only nonpacemakers were able to produce networkwide bursting, but with a low probability of bursting and low input and output ranges. Our results indicate that inclusion of pacemakers in an excitatory network increases robustness of the network by more than tripling the input and output ranges compared with networks containing no pacemakers. The largest increase in dynamic range occurs when the number of pacemakers in the network is greater than 20% of the population. Experimental tests of our model predictions are proposed.

Afferent Pathways↗
PubMed2003

Holographic common-path interferometer for angular displacement measurements with spatial phase stepping and extended measurement range.

A novel technique for extending the unambiguous measurement range for differential measurements of angular deflections is presented. The technique utilizes a common-path interferometer that simultaneously probes the out-of-plane displacement of three points on the object surface. The system is based on a single laser diode, and all the optical functions of the system are implemented in a dedicated holographic optical element (HOE). The HOE automatically provides spatially phase-stepped interference signals for real-time phase measurement. It is therefore not necessary to employ any polarizing optics or active elements to introduce the phase stepping. The common-path scheme combined with the HOE provides a system that is inherently stable, since the HOE operates as both transmitter and receiver in the system. The system is compact, is robust, and has the potential for being mass-produced at a low cost and is thus well suited for industrial use, such as in commercial vibrometers. The technique is demonstrated in a system for measuring angular deflections of a plane mirror. The technique, however, is not restricted to this use alone and can easily be configured to probe other types of surface displacements, e.g., the deflection of a diaphragm. In the present configuration, the system can measure angular deflections with a sensitivity of 2.5 x 10(-7) rad over a measurement range that is approximately 3.5 x 10(-3) rad, i.e., a dynamic range of approximately 1:14,000. Furthermore, the system can easily be reconfigured for a desired angular sensitivity and measurement range.

Journal Article↗
PubMed1980

Enhancement of the Yu and Ledeen gas-liquid chromatographic method for sialic acid estimation: use of methane chemical ionization mass fragmentography.

The sialic acid present in erythrocyte ghosts was estimated by methane chemical ionization mass fragmentography of the trimethylsilyl methyl glycosides. The internal standard was 3,4,6-tris-trimethylsilyl-alpha-phenyl-2-deoxy-2-acetamide-D-glucosaminide, as proposed by Yu and Ledeen (J. Lipid Res. 1970. 11: 506-516). The [MH-16]+ions (m/e 610 for the TMS-methylglycoside of N-acetylneuraminic acid and m/e 498 for the internal standard) were used for quantifying nanogram levels of sialic acid in the presence of other contaminating substances. This effectively raises the signal-to-noise ratio for the Yu and Ledeen method by at least two orders of magnitude. The sensitivity and linearity of the method, without use of isotopic carriers, were tested using known quantities of N-acetylneuraminic acid. The limit of detection was below 0.4 nanograms (approximately one picomole). The useful range of detection was 10 ng-1 microgram, showing a large dynamic range.

Erythrocyte Membrane↗
PubMed2003

The sciatic nerve of the toad Xenopus laevis as a physiological model of the human cochlear nerve.

The response of single fibres of the human cochlear nerve to electrical stimulation by a cochlear implant has previously been inferred from the response of the cochlear nerve in other mammals. These experiments are hindered by stimulus artefact and the range of stimulus currents used is therefore much less than the perceptual dynamic range (from threshold to discomfort) of human subjects. We have investigated use of the sciatic nerve of the toad Xenopus laevis as a convenient physiological model of the human cochlear nerve. Use of this completely dissected nerve reduces the problems of stimulus artefact whilst maintaining the advantages of a physiological preparation. The validity of the model was assessed by measuring the refractory periods, excitation time-constant, and relative spread of single fibres using microelectrode recording. We have also investigated the response of nerve fibres to sinusoidal stimulation. Based on these measurements, we propose that the sciatic nerve may be a suitable model of the human cochlear nerve if the timescales of stimuli are decreased by a factor of about five to compensate for the slower dynamics of the sciatic nerve and if noise is added to the stimuli to compensate for the lower internal noise of sciatic nerve fibres.

Animals↗
PubMed1996

[Effect of mutations on the dynamics of numbers of individuals with different genotypes in an isolated heterogeneous population].

The overall (throughout the range) dynamics of the number of diploid individuals possessing n possible gene alleles in a given autosomal locus is studied in an isolated heterogeneous population. The significant influence of new mutations on overall dynamics is shown. It is demonstrated that, under certain conditions, the selection mechanism might have a greater effect on the dynamics of number of individuals with different genotypes in the isolated heterogeneous population than on mutation and "death-reproduction" mechanisms.

Genetic Heterogeneity↗