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Biomedical subjects

B E Chapman

Publications and source records attributed to B E Chapman.

At least 19 recordsLinked to original sources

NMR measurements of the diffusional permeability of water in cultured colonic epithelial cancer cells.

The water residence time and diffusional water permeability in colonic epithelial T84 cancer cells was measured using (1)H NMR spectroscopy; the values estimated were 35.2+/-2.8 ms and (7.4+/-0.6)x10(-3)cms(-1), respectively. Water permeability was inhibited to approximately 10% of its original value by the mercurial diuretic, p-chloromercuribenzenesulfonate (PCMBS; 1mM), and fully restored by dithiothreitol (DTT; 1mM). The permeability was also inhibited reversibly to approximately 55%, by extracellular glibenclamide (1mM), an inhibitor of some ATP-binding cassette (ABC) transporters, including the cystic fibrosis transmembrane conductance regulator (CFTR). Addition of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IMBX; 0.1-1mM) and the adenylate cyclase activator, forskolin (0.1-1mM) did not alter water permeability. It is concluded that in T84 cells water diffuses through the membrane lipid bilayer and via channels that are inhibited by PCMBS, including the channels that are known to be inhibited by glibenclamide.

4-Chloromercuribenzenesulfonate↗

Observations of multiple magnetic islands in the core of a reversed field pinch.

We describe in this Letter the first measurement of multiple islands in the core of a reversed field pinch (RFP). These islands appear with current profile modification leading to magnetic fluctuation reduction in the Madison symmetric torus RFP. Magnetic island widths decrease to an unprecedented level, reducing the overlap of adjacent islands and allowing distinct islands to appear. The structures are observed in multichord measurements of soft-x-ray emissivity. The soft-x-ray data is validated with Poincaré reconstructions of the magnetic field structure in the core.

Journal Article↗

31P MAS-NMR of human erythrocytes: independence of cell volume from angular velocity.

31P magic angle spinning NMR (MAS-NMR) spectra were obtained from suspensions of human red blood cells (RBCs) that contained the cell-volume-sensitive probe molecule, dimethyl methylphosphonate (DMMP). A mathematical representation of the spectral-peak shape, including the separation and width-at-half-height in the 31P NMR spectra, as a function of rotor speed, enabled us to explore the extent to which a change in cell volume would be reflected in the spectra if it occurred. We concluded that a fractional volume change in excess of 3% would have been detected by our experiments. Thus, the experiments indicated that the mean cell volume did not change by this amount even at the highest spinning rate of 7 kHz. The mean cell volume and intracellular 31P line-width were independent of the packing density of the cells and of the initial cell volume. The relationship of these conclusions to other non-NMR studies of pressure effects on cells is noted.

Computer Simulation↗

Observation of velocity-independent electron transport in the reversed field pinch.

Confinement of runaway electrons has been observed for the first time in a reversed field pinch during improved-confinement plasmas in the Madison Symmetric Torus. Energy-resolved hard-x-ray flux measurements have been used to determine the velocity dependence of the electron diffusion coefficient, utilizing computational solutions of the Fokker-Planck transport equation. With improved-confinement, the fast electron diffusivity drops by 2 orders of magnitude and is independent of velocity. This suggests a change in the transport mechanism away from stochastic magnetic field diffusion.

Journal Article↗

Comparative NMR studies of diffusional water permeability of red blood cells from different species: XIV. Little Penguin, Eudyptula minor.

As part of a programme of comparative measurements of diffusional water permeability (Pd) the red blood cells (RBC) from Little Penguin (Eudyptula minor) were studied. The cell dimensions were measured with light and electron microscopy, and by a newly described non-invasive technique, NMR q-space analysis. In view of its relative novelty for cell biologists, an overview of this technique is presented. The RBC revealed an ellipsoidal shape that is characteristic of avian RBC, with axis lengths ("diameters") estimated to be: a=16.0 microm; b=9.6 microm; c=5.0 microm. The values of P(d)were: 2.0 x 10(-3)cm s(-1)at 5 degrees C, 3.3 x 10(-3)cm s(-1)at 10 degrees C, 4.6 x 10(-3)cm s(-1)at 15 degrees C and approximately 5.4 x 10(-3)cm s(-1)at 20, 25, 30, 37 and 42 degrees C. There was a lack of inhibition of water permeability by p-chloromercuribenzensulfonate (PCMBS), the well-known inhibitor of RBC aquaporin. It was notable that in the temperature range 5-20 degrees C the NMR parameters, and hence the permeability, varied linearly as is found for other species, but at temperatures higher than 20 degrees C there was no temperature-dependence of Pd. Consequently, there was an obvious break at approximately 20 degrees C in the Arrhenius plot, of the mean residence life time of water inside the cells, 1/Te, versus temperature. For temperatures less than 20 degrees C the activation energy E(a,d) was 45.6 +/- 6.6 kJ/mol. For temperatures higher than 25 degrees C E(a,d) was zero. The lack of inhibition of water permeability by PCMBS and the very high value of E(a,d) for diffusive water exchange suggests that the water permeation occurs primarily via the membrane bilayer per se, i.e., there is no aquaporin in Little Penguin RBC. The discontinuity at approximately 20 degrees C in the Arrhenius plot is an interesting finding, not seen before in other species, and we suggest that it reflects a phase transition in the membrane lipids.

4-Chloromercuribenzenesulfonate↗

Effects of p-chloromercuribenzene sulfonate on water transport across the marsupial erythrocyte membrane.

The effects of exposure of red blood cells (RBC) of three species of marsupial to a mercury-containing sulfhydryl-modifying reagent, p-chloromercuribenzene sulfonate (PCMBS), on the water diffusional permeability ( P (d)) of their membranes were monitored by using an Mn(2+)-doping (1)H nuclear magnetic resonance (NMR) technique at 400 MHz. For koala ( Phascolarctos cinereus), RBC the maximal inhibition was reached at 37 degrees C in 60 min with 1 mmol.l(-1) PCMBS or in 15-30 min with 2 mmol. l(-1) PCMBS. In contrast, in the case of red kangaroo ( Macropus rufus) or swamp wallaby ( Wallabia bicolor) RBC, maximal inhibition required an incubation of 90 min at 37 degrees C with 2 mmol.l(-1) PCMBS. For the RBC of all three species the value of maximal inhibition was very high, being 50-70% when measured at 25 degrees C, 60-80% at 30 degrees C and 60-70% at 37 degrees C. The lowest values of P (d) appeared to be around 2 x 10(-3)-3 x 10(-3) cm.s(-1) in the temperature range of 25-37 degrees C. The mean value of the activation energy of water diffusion ( E (a,d)) was approximately 20-25 kJ.mol(-1) for control and approximately 40 kJ.mol(-1) for PCMBS-inhibited RBCs. These results show that marsupial RBC have a basal permeability to water similar to that previously reported for human RBC, but a higher value of the PCMBS-inhibitable water permeability. This indicates that the higher water permeability of marsupial RBC compared with human RBC is associated with a higher fraction of protein-mediated water permeability.

4-Chloromercuribenzenesulfonate↗

Measurement of the current-density profile and plasma dynamics in the reversed-field pinch.

First measurements of the current-density profile in the core of a high-temperature reversed-field pinch are presented. The current-density profile is observed to peak during the sawtooth cycle and broaden promptly at the crash. This change in profile can be linked to magnetic relaxation and the dynamo which is predicted to drive antiparallel current in the plasma core. For high-confinement discharges, the dynamo is suppressed and the current-density profile is observed to strongly peak.

Journal Article↗

Reduced edge instability and improved confinement in the MST reversed-field pinch.

Improved confinement has been achieved in the MST through control of the poloidal electric field, but it is now known that the improvement has been limited by bursts of an edge-resonant instability. Through refined poloidal electric field control, plus control of the toroidal electric field, we have suppressed these bursts. This has led to a total beta of 15% and a reversed-field-pinch-record estimated energy confinement time of 10 ms, a tenfold increase over the standard value which for the first time substantially exceeds the confinement scaling that has characterized most reversed-field-pinch plasmas.

Journal Article↗

A comparison of classification algorithms to automatically identify chest X-ray reports that support pneumonia.

We compared the performance of expert-crafted rules, a Bayesian network, and a decision tree at automatically identifying chest X-ray reports that support acute bacterial pneumonia. We randomly selected 292 chest X-ray reports, 75 (25%) of which were from patients with a hospital discharge diagnosis of bacterial pneumonia. The reports were encoded by our natural language processor and then manually corrected for mistakes. The encoded observations were analyzed by three expert systems to determine whether the reports supported pneumonia. The reference standard for radiologic support of pneumonia was the majority vote of three physicians. We compared (a) the performance of the expert systems against each other and (b) the performance of the expert systems against that of four physicians who were not part of the gold standard. Output from the expert systems and the physicians was transformed so that comparisons could be made with both binary and probabilistic output. Metrics of comparison for binary output were sensitivity (sens), precision (prec), and specificity (spec). The metric of comparison for probabilistic output was the area under the receiver operator characteristic (ROC) curve. We used McNemar's test to determine statistical significance for binary output and univariate z-tests for probabilistic output. Measures of performance of the expert systems for binary (probabilistic) output were as follows: Rules--sens, 0.92; prec, 0.80; spec, 0.86 (Az, 0.960); Bayesian network--sens, 0.90; prec, 0.72; spec, 0.78 (Az, 0.945); decision tree--sens, 0.86; prec, 0.85; spec, 0.91 (Az, 0.940). Comparisons of the expert systems against each other using binary output showed a significant difference between the rules and the Bayesian network and between the decision tree and the Bayesian network. Comparisons of expert systems using probabilistic output showed no significant differences. Comparisons of binary output against physicians showed differences between the Bayesian network and two physicians. Comparisons of probabilistic output against physicians showed a difference between the decision tree and one physician. The expert systems performed similarly for the probabilistic output but differed in measures of sensitivity, precision, and specificity produced by the binary output. All three expert systems performed similarly to physicians.

Acute Disease↗

Quantifying the characteristics of unambiguous chest radiography reports in the context of pneumonia.

RATIONALE AND OBJECTIVES: The purpose of this study was to statistically identify some characteristics of unambiguous (ie, clear) chest radiography reports in the context of acute bacterial pneumonia. MATERIALS AND METHODS: Seven physicians individually read 292 chest radiography reports to determine if they contained radiologic evidence of pneumonia. Unambiguous reports were defined as those that physicians unanimously classified as supporting or not supporting the diagnosis of pneumonia. Ambiguous reports were assigned degrees of ambiguity on the basis of how much disagreement they caused among the physicians. Characteristics of unambiguous reports as described in the literature were manually quantified and assigned to every report. To identify characteristics that statistically distinguished unambiguous from ambiguous reports, the authors performed an ordinal logistic regression analysis for which the dependent variable was the number of dissenting votes the report received and the independent variables were the quantified characteristics of the report. RESULTS: Six independent variables were statistically significantly associated with unambiguous reports (P < .05). Three were positively associated: an interpretation of whether findings supported the diagnosis of pneumonia in reports with pneumonia-related observations, short sentences, and redundancy of pneumonia-related observations. Three were negatively associated: high use of uncertainty modifiers for pneumonia-related observations, use of only descriptive terms to describe pneumonia-related observations, and insufficient amount of pneumonia-related information. CONCLUSION: The most influential characteristic of an unambiguous chest radiography report was an interpretation of whether the radiograph supported the diagnosis of pneumonia when findings could be indicative.

Diagnosis, Differential↗

Determination of NADH-dependent glutamate synthase (GOGAT) in Spodoptera frugiperda (Sf9) insect cells by a selective 1H/15N NMR in vitro assay.

This is the second of two papers [Drews, M., Doverskog, M., Ohman, L., Chapman, B.E., Jacobsson, U., Kuchel, P.W., Häggström, L., 2000. Pathways of glutamine metabolism in Spodoptera frugiperda (Sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1H/15N NMR. J. Biotechnol. 78, 23-37]. where the general goal has been to determine and characterise the glutamine metabolism in Sf9 cells. The presence of glutamate synthase (GOGAT) activity was investigated in cell-free extracts of S. frugiperda (Sf9) insect cells by modified 1H/15N spin-echo and gradient enhanced multiple quantum coherence NMR spectroscopy techniques. Cell-free extracts were prepared from cells cultured in a serum-free medium. The assay conditions were based on conventional spectrophotometric and chromatographic methods. NMR data showed that nitrogen from [5-15N] glutamine was selectively incorporated into 2-oxoglutarate forming [2-15N] glutamate with a specific activity of 4.15 +/- 0.21 nmol [2-15N] glutamate min -1 (mg total protein)-1 in the cell-free extracts. The enzyme activity was exclusively dependent on NADH as coenzyme and was completely inhibited by 1 mM azaserine. From the results obtained, we conclude that Sf9 cells possess NADH-GOGAT activity. Furthermore, the high specificity of the NMR method enables distinction of competing reactions from glutaminase and glutamate dehydrogenase.

Animals↗

Pathways of glutamine metabolism in Spodoptera frugiperda (Sf9) insect cells: evidence for the presence of the nitrogen assimilation system, and a metabolic switch by 1H/15N NMR.

1H/15N and 13C NMR were used to investigate metabolism in Spodoptera frugiperda (Sf9) cells. Labelled substrates ([2-15N]glutamine, [5-15N]glutamine, [2-15N]glutamate, 15NH4Cl, [2-15N]alanine, and [1-13C]glucose) were added to batch cultures and the concentration of labelled excreted metabolites (alanine, NH4+, glutamine, glycerol, and lactate) were quantified. Cultures with excess glucose and glutamine produce alanine as the main metabolic by-product while no ammonium ions are released. 1H/15N NMR data showed that both the amide and amine-nitrogen of glutamine was incorporated into alanine in these cultures. The amide-nitrogen of glutamine was not transferred to the amine-position in glutamate (for further transamination to alanine) via free NH4+ but directly via an azaserine inhibitable amido-transfer reaction. In glutamine-free media 15NH4+ was consumed and incorporated into alanine. 15NH4+ was also incorporated into the amide-position of glutamine synthesised by the cells. These data suggest that the nitrogen assimilation system, glutamine synthetase/glutamate synthase (NADH-GOGAT), is active in glutamine-deprived cells. In cultures devoid of glucose, ammonium is the main metabolic by-product while no alanine is formed. The ammonium ions stem both from the amide and amine-nitrogen of glutamine, most likely via glutaminase and glutamate dehydrogenase. 13C NMR revealed that the [1-13C] label from glucose appeared in glycerol, alanine, lactate, and in extracellular glutamine. Labelling data also showed that intermediates of the tricarboxylic acid cycle were recycled to glycolysis and that carbon sources, other than glucose-derived acetylCoA, entered the cycle. Furthermore, Sf9 cell cultures excreted significant amounts glycerol (1.9-3.2 mM) and ethanol (6 mM), thus highlighting the importance of sinks for reducing equivalents in maintaining the cytosolic redox balance.

Alanine↗

Enhanced image detail using continuity in the MIP Z-buffer: applications to magnetic resonance angiography.

In this paper a new algorithm is presented for the segmentation and display of blood vessels from images obtained with magnetic resonance angiography (MRA) and other three-dimensional (3D) angiographic imaging techniques. The algorithm developed is based on the observation that vessels are strongly evident in the maximum intensity projection (MIP) Z-buffer as regions of high continuity and low local roughness. Roughness is measured here by the minimum chi2 value of a low-order local least-squares fit in the principal directions through each point in the MIP Z-buffer. Although some background pixels in the Z-buffer exhibit low local roughness, the size of the connected region is nearly always much smaller than even the very smallest vessels that appear in the MIP image. It is shown that by applying connectivity to the regions of low roughness, there is nearly complete separation between vascular detail and background. When connectivity is further applied in the original 3D image, vascular bed segmentation becomes nearly complete. The algorithm consists of three basic steps: a) determination of the minimum local roughness at each point in the MIP Z-buffer; b) connection of all neighboring points of low local roughness; and c) connection of all points in the original 3D image matrix that are connected to the points determined in the MIP Z-buffer and that are above an intensity threshold. The algorithm as presented is not optimized but demonstrates a very strong potential for improved portrayal of vascular detail.

Algorithms↗

A three-coil comparison for MR angiography.

The purpose of this work was to compare intracranial magnetic resonance angiography (MRA) image quality using three different radiofrequency coils. The three coil types included a reduced volume quadrature birdcage coil with endcap, a commercially available quadrature birdcage head coil, and a four-element phased-array coil. Signal-to-noise ratio (SNR) measurements were obtained from comparison studies performed on a uniform cylindrical phantom. MRA comparisons were performed using data acquired from 15 volunteers and applying a thick-slab three-dimensional time-of-flight sequence. Analysis was performed using the signal difference-to-noise ratio, a quantitative measure of the relative vascular signal. The reduced-volume endcap and phased-array coils, which were designed specifically for imaging the intracranial volume of the head, improved the image SNR and vascular detail considerably over that obtained using the commercially available head coil. The endcap coil configuration provided the best vascular signal overall, while the phased-array coil provided the best results for arteries close to the coil elements.

Brain↗

A median filter for 3D FAST spin echo black blood images of cerebral vessels.

High-resolution black-blood MRA images of intracranial vascular anatomy can be obtained using 3D fast spin-echo techniques. Although these images demonstrate excellent contrast between vessels and surrounding soft tissues, the dark signal from air and bone can obscure the desired vascular information when a minimum intensity projection image is created. In this paper, we describe an image processing technique based upon a median filter that is effective for detecting narrow vessel-like structures. Minimum intensity projection images of the filtered MRA volume can be obtained in any orientation without prior segmentation of the skull or surrounding air spaces. The filter is very effective for detecting and visualizing small vessels, but is much less effective for detecting vessels and vascular pathology larger than the filter detection width. The filtering technique is demonstrated on black-blood MRA images from a volunteer study.

Algorithms↗

Evidence of red cell alignment in the magnetic field of an NMR spectrometer based on the diffusion tensor of water.

The alignment of human erythrocytes in aqueous suspensions in the magnetic field B(0) (called the z-direction) of an NMR spectrometer was shown by calculating the diffusion tensor for water in the sample. The diffusion was measured using a pulsed-field-gradient spin-echo NMR method. The extent of diffusion anisotropy for water was exemplified by the values of the apparent diffusion coefficients with erythrocytes of normal shape and volume: for a typical experiment the values for the x-, y-, and z-directions were (6.88 +/- 0.17) x 10(-10), (7.07 +/- 0.17) x 10(-10), and (10.20 +/- 0.17) x 10(-10) m(2) s(-1), respectively. Cells in hypo- and hyperosmotic media were also studied and they too showed the anisotropy of the apparent diffusion coefficients but the extents were different. A new method of data analysis was developed using the Standard Add-On Packages in a Mathematica program. The experimental findings support evidence of erythrocyte alignment that was previously obtained with a high-field-gradient q-space method.

Diffusion↗

Parallel secretion of pancreatic phospholipase A(2), phospholipase A(1), lipase, and colipase in children with exocrine pancreatic dysfunction.

The cosecretion of pancreatic lipase and colipase are important in normal fat digestion. As adsorption of phosphatidylcholine to the lipid substrate interferes with lipase activity, hydrolysis to lysophosphatidylcholine with subsequent desorption is also essential for fat digestion. There are some data regarding the secretion of pancreatic phospholipases in normal adults but none in children or patients with pancreatic disease. In the present study, we aimed a) to develop an accurate fast assay method to measure phospholipase A(2) and b) to determine the secretion rate of pancreatic phospholipase A(2) and whether it is cosecreted with lipase and colipase in children with exocrine pancreatic dysfunction. Nine male patients aged 0.5 to 16 y (seven with cystic fibrosis, two with malabsorption) underwent pancreatic stimulation tests. Their colipase and lipase secretion rates were measured by titrimetric methods and phospholipase A(2) and A(1) by phosphorus magnetic resonance spectroscopy ((31)P NMR). It was found that the phospholipases, colipase, and lipase were absent in the two patients with pancreatic insufficiency. In patients with normal absorption, there were marked inter-and intrasubject variations of lipase, colipase, and phospholipase secretion rates that were consistent with the degree of exocrine pancreatic dysfunction. However, in the three 20-min stimulation periods of the pancreatic function test, pancreatic phospholipase is cosecreted with lipase and colipase, and average colipase and phospholipase A(2) secretion rates follow a similar or parallel pattern. These findings are consistent with the important role of pancreatic phospholipases in intestinal phospholipid hydrolysis leading to the desorption of phospholipids from the lipid substrate and enhancing lipid hydrolysis and phospholipid absorption.

Adolescent↗

Evaluation of measures of technical image quality for intracranial magnetic resonance angiography.

We evaluate three measures of technical image quality for intracranial magnetic resonance angiography (MRA): (1) a two-alternative forced choice (2AFC) evaluation of vessel visibility, (2) vessel-to-background signal-difference-to-noise ratio (SDNR), and (3) observer ranking of the fidelity of vessel morphology compared to that in a gold standard image. The gold standard used for both the 2AFC and ranking measures is intraarterial catheter angiography. These measures are applied to healthy arterial segments. The 2AFC and SDNR measures directly evaluate the visibility of artery segments for which the existence is known from the gold standard images. We argue that (1) 2AFC evaluates the carrier signals on which any vascular disease process is modulated and provides an upper bound on the detectibility of vascular lesions, (2) SDNR is a predictor of 2AFC, and (3) ranking may be used to predict the relative performance of techniques in the detection of vascular lesions.

Adult↗