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Biomedical subjects

K A Powell

Publications and source records attributed to K A Powell.

18 recordsLinked to original sources
PubMed1997

Compartment-ablation studies of GLUT4 distribution in adipocytes: evidence for multiple intracellular pools.

The available data suggest that GLUT4 does populate the recycling endosomal system to some extent, but that a large proportion of the intracellular GLUT4 resides in a compartment that is devoid of transferrin receptors and may have properties more akin to specialized secretory vesicles. The study of the nature and biogenesis of this compartment will provide important insight into the mechanism by which insulin stimulates glucose transport. Further study of the role of the synaptobrevins in these distinct subcellular compartments will probably shed further light on the mechanism by which insulin stimulates GLUT4 translocation.

Adipocytes↗
PubMed1996

Quantitative detection of outlet strut separations in Björk-Shiley convexo-concave mitral valves.

BACKGROUND: As of January 31, 1995, 564 outlet strut fractures (OSFs) of Björk-Shiley convexo-concave (BSCC) heart valves had been reported to the Shiley Heart Valve Research Center, of which approximately two thirds resulted in the death of the patient. Previous studies indicate that one leg of the outlet strut separates (single-leg separation, SLS) from the valve before the second leg breaks, which results in complete OSF. To identify those valves at risk of complete OSF, an in vivo radiographic imaging technique is being developed to evaluate the strut leg integrity. The goal of the present study was to develop an objective postprocessing technique to evaluate outlet strut leg integrity quantitatively in these cineradiographic images. METHODS AND RESULTS: Twenty-two sets (12 intact valves, 10 SLS valves) of cineangiographic images were obtained from individuals whose valve status was subsequently verified ex vivo. Several quantitative measures of SLS were evaluated to identify possible loss of metal or gaps in the SLS legs. Two of these measures, decrease in pixel intensity (DIPI) ratio and gap half-width, are diagnostic metrics of SLS: i.e., the maximum likelihood estimate of the area under the receiver operating characteristic curve was 0.892 (SD, 0.066) for a model based on DIPI ratio and 0.802 (SD, 0.093) for gap half-width. CONCLUSIONS: We have developed a postprocessing technique that can be used to objectively evaluate outlet strut integrity in cineradiographic images of BSCC heart valves. At an estimated specificity of 1.0, the estimated sensitivity of the objective review was comparable to that of a subjective expert review panel.

Analysis of Variance↗
PubMed1996

Dynamic magnetic resonance imaging assessment of the effect of ventricular wall curvature on regional function in hypertrophic cardiomyopathy.

We hypothesized that contraction within the ventricular septum in hypertrophic cardiomyopathy (HC) may be related to its abnormal morphology because ventricular wall stress is related to wall curvature by the Laplace equation. To test this, we studied 17 HC patients with various septal morphologies using dynamic magnetic resonance imaging techniques. Short- and long-axis curvatures of the basal septal and basal lateral walls were determined on cine images as the reciprocal of the radius of the arc best fit to the endocardial contour, which was negative if the wall was convex to the cavity of the left ventricle. Endocardial and epicardial intramyocardial circumferential shortening (% circumferential shortening) was measured in the septal and lateral walls on basal short-axis myocardial tagging images. Septal walls were flatter in the short-axis plane and more convex toward the left ventricular cavity in the long-axis plane than lateral walls, as indicated by smaller short- and long-axis curvatures. Septal percent circumferential shortening was significantly lower than the lateral percent circumferential shortening, suggesting reduced septal contraction. Endocardial and epicardial percent circumferential shortening showed significant positive correlations with wall curvatures. Multiple stepwise linear regression analysis revealed that both short- and long-axis curvatures significantly contributed to percent circumferential shortening (r=0.87 for endocardial and r=0.70 for epicardial, both p<0.0001). In conclusion, wall curvature is related to wall function in HC; the more convex toward the left ventricular cavity the wall is, the less it contracts. Reduced contraction of the septum in HC may be partly due to its abnormal curvature.

Cardiomyopathy, Hypertrophic↗
PubMed1996

Impact of wall constraint on velocity distribution in proximal flow convergence zone. Implications for color Doppler quantification of mitral regurgitation.

OBJECTIVES: This study sought to elevate the effect of proximal flow constraint induced by the left ventricular wall on the accuracy of calculated flow rates and to assess a possible correction factor to adjust the proximal convergence angle. We further defined under which hydrodynamic and geometric conditions it is necessary to apply the corrected convergence angle. BACKGROUND: The proximal flow convergence method has been proposed as a new approach to quantify valvular regurgitation. However, significant overestimation of the calculated regurgitant flow rate has been reported, particularly in patients with mitral valve prolapse and severe mitral regurgitation. METHODS: We used an in vitro flow model and induced various degrees of proximal flow constraint. The accuracy of the proposed convergence angle formula, alpha = tau + 2 tan-1 d/r (d = wall distance; r = isovelocity radius) was tested in vitro and in a three-dimensional numerical simulation. RESULTS: With a constraining wall near the orifice, overstimulation of regurgitant flow rates was noted and was most significant with the constraining wall positioned closest to the orifice (calculated flow rate [Qc]/true flow rate [Qo] = 1.85 +/- 0.55 [mean +/- SD]). These findings were similar to the results of the numerical simulation. Applying the correction factor nearly completely eliminated the overestimation of the calculated flow rates (cQc), with cQc/Qo = 1.13 +/- 0.25. CONCLUSIONS: In the presence of a constraining wall, significant overestimation of calculated flow rates is observed when hemispheric symmetry of the flow field is assumed. In this situation, it is necessary to apply the corrected convergence angle formula to improve the accuracy of the proximal flow convergence method.

Bias↗
PubMed1996

Quantitative classification of breast tumors in digitized mammograms.

The goal of this study was to develop a technique to distinguish benign and malignant breast lesions in secondarily digitized mammograms. A set of 51 mammograms (two views/patient) containing lesions of known pathology were evaluated using six different morphological descriptors: circularity, mu R/sigma R (where mu R = mean radial distance of tumor boundary, sigma R = standard deviation); compactness, P2/A (where P = perimeter length of tumor boundary and A = area of the tumor); normalized moment classifier; fractal dimension; and a tumor boundary roughness (TBR) measurement (the number of angles in the tumor boundary with more than one boundary point divided by the total number of angles in the boundary). The lesion was segmented from the surrounding background using an adaptive region growing technique. Ninety-seven percent of the lesions were segmented using this approach. An ROC analysis was performed for each parameter and the results of this analysis were compared to each other and to those obtained from a subjective review by two board-certified radiologists who specialize in mammography. The results of the analysis indicate that all six parameters are diagnostic for malignancy with areas under their ROC curves ranging from 0.759 to 0.928. We observed a trend towards increased specificity at low false-negative rates (0.01 and 0.001) with the TBR measurement. Additionally, the diagnostic accuracy of a classification model based on this parameter was similar to that of the subjective reviewers.

Breast Diseases↗
PubMed1995

Pressure recovery in bileaflet heart valve prostheses. Localized high velocities and gradients in central and side orifices with implications for Doppler-catheter gradient relation in aortic and mitral position.

BACKGROUND: We investigate pressure recovery in central and side orifices of St Jude valves and the effect of mitral versus aortic position on the relation between Doppler- and catheter-derived pressure gradients. METHODS AND RESULTS: Maximum, transvalvular, and net pressure gradients are calculated and compared with Doppler-derived gradients in an in vitro model. Pressure recovery and pressure loss coefficients are calculated. Simultaneous Doppler and catheter gradients are obtained intraoperatively in five patients undergoing mitral valve replacement. Centerline Doppler gradients correspond closely with maximum catheter gradients but are higher than transvalvular and net pressure gradients. Thirty-six percent of the initial pressure drop is recovered between the valve leaflets and is independent of valve size or configuration. A variable amount of postvalvular pressure recovery is observed depending on aortic or mitral configuration. Side orifice velocities are 85 +/- 4% of the centerline velocities. Incorporation of the pressure loss coefficient in the simplified Bernoulli equation shows close agreement between centerline Doppler gradients and transvalvular gradients (r = .99, y = 1.11x-0.19). CONCLUSIONS: Gradients across the St Jude valve measured by Doppler ultrasound are higher than transvalvular or net catheter gradients due to downstream pressure recovery. This is more marked for Doppler gradients based on centerline velocities than side orifice velocities and is more pronounced for valves in an aortic than a mitral configuration. Therefore, to be comparable with invasive transvalvular catheter gradients, either Doppler gradients should be calculated based on side orifice velocity measurements or the Doppler gradient calculation should include the pressure loss coefficient when based on central orifice velocities.

Aortic Valve↗
PubMed1995

Dephosphin/dynamin is a neuronal phosphoprotein concentrated in nerve terminals: evidence from rat cerebellum.

Dephosphin/dynamin is a 94,000/96,000 mol. wt protein kinase C substrate from rat brain that is phosphorylated in nerve terminals and dephosphorylated upon stimulation of exocytosis and synaptic vesicle recycling. Phosphorylation activates an intrinsic GTPase activity and dephosphin may play a role in endocytosis [Robinson P. J. et al. (1993) Nature 365, 163-166]. In this study a specific polyclonal antiserum to dephosphin was used to investigate its distribution in rat brain by immunoblotting and immunocytochemistry. Immunoblots of various organs of the rat showed that dephosphin was detectable only in the whole brain and not in the testes, lung, kidney, adrenals, heart, liver or skeletal muscle. Immunoblotting of various regions of the brain revealed high levels of dephosphin, particularly in the hippocampus, cerebellum and cerebral cortex, but its absence from the anterior pituitary. Synaptosomes were prepared from these three regions and labelled with 32Pi for 60 min, followed by incubation in control or 41 mM K+ depolarizing buffer. Dephosphin was present in each region and was stoichiometrically dephosphorylated by depolarization, indicating the presence and regulation of dephosphin in intact cerebellar nerve terminals. The cerebellum was selected for detailed study, using conventional light and confocal microscopy, owing to its ordered and well-characterized structure. Immunostaining was abundant within the cerebellar cortex and deep cerebellar nuclei, but almost entirely absent from the medulla. In the cortex many neuronal cells contained dephosphin-like immunoreactivity which was also evident in perikarya, axons, and nerve terminals. Dephosphin-like immunoreactivity was not detected in the radial Bergman glial cells. The greatest concentrations were observed in synaptic terminals, particularly in granular layer glomeruli and basket cell terminals surrounding Purkinje cell bodies and dendrites. Dephosphin therefore appears to be exclusive to neuronal tissue, but is distributed widely throughout the brain. It is located in many neuronal cell types of the cerebellum and may be particularly enriched in synaptic terminals, where it is regulated by phosphorylation and dephosphorylation. This distribution suggests a role for dephosphin in synaptic vesicle cycling in nerve terminals.

Alkaline Phosphatase↗
PubMed1994

Dynamin I is a Ca(2+)-sensitive phospholipid-binding protein with very high affinity for protein kinase C.

Depolarization-induced Ca2+ influx into rat brain synaptosomes induces dephosphorylation of dephosphin, a 96-94-kDa protein kinase C (PKC) substrate recently identified as dynamin I, a protein associated with endocytosis. We characterized purified dynamin I to better understand regulation of its phosphorylation in nerve terminals. Purified dynamin I possessed a very high affinity for PKC but did not fit Michaelis-Menten kinetics. It had an optimum phosphorylation rate of 1.42 +/- 0.02 mumol/mg/min and a concentration giving half-maximal activity (S0.5) of 0.14 +/- 0.02 microM, the highest affinity reported for a PKC substrate protein. Concentrations of dynamin greater than 0.5 microM inhibited phosphorylation. The stoichiometry was 1.5, indicating more than one phosphorylation site. Dynamin was predominantly associated with the brain particulate fraction under conditions of low ionic strength, and this prevented its phosphorylation by PKC until released by moderate increases in ionic strength (Na+, K+, and Mg2+) or by GTP or ATP. In intact synaptosomes the largest dynamin pool was associated with the particulate fraction, while a smaller pool was cytosolic or extracted with 150 nM NaCl and contained all the phosphorylated protein. Purified dynamin also bound to phospholipid-coated controlled-pore glass beads, but poorly in the presence of NaCl, Mg2+, GTP, or ATP. Ca2+ induced a reversible translocation from the cytosol to the particulate fraction (50% at 183 microM Ca2+) in brain homogenates, and the purified protein also underwent Ca(2+)-sensitive translocation to phospholipid-coated controlled-pore glass beads. We conclude that dynamin I is a nerve terminal Ca(2+)-sensitive phospholipid-binding protein with very high substrate affinity for PKC. We propose that phosphorylation by PKC occurs in the nerve terminal soluble compartment and that Ca2+ may mediate its binding to the particulate fraction, thereby blocking the PKC phosphorylation sites. These properties may contribute to the lack of PKC phosphorylation during depolarization, despite the presence of activated PKC.

Amino Acid Sequence↗
PubMed1994

Phosphorylation of dynamin I and synaptic-vesicle recycling.

In nerve terminals, neurotransmitters are packaged in synaptic vesicles, and released by exocytosis. Empty synaptic vesicles are rapidly recycled for reuse by endocytosis. Much progress has been made in identifying the proteins involved in synaptic-vesicle trafficking, but the mechanism and regulation of endocytosis have largely remained an enigma. One approach to defining regulatory proteins that might be involved is to study stimulus-dependent phosphorylation events in nerve terminals. This has led to the identification of dephosphin, which is quantitatively dephosphorylated by nerve-terminal depolarization. Sequencing reveals that dephosphin is identical with dynamin I, a GTP-binding protein that functions in endocytosis. Phosphorylation and dephosphorylation of nerve-terminal dynamin I/dephosphin regulates its intrinsic GTPase activity in parallel with the regulation of synaptic-vesicle recycling. Therefore, phosphorylation and dephosphorylation of dynamin I might provide a Ca(2+)-dependent switch for endocytosis in the synaptic-vesicle pathway.

Animals↗
PubMed1988

Structural studies of nerve terminals containing melanin-concentrating hormone in the eel, Anguilla anguilla.

Eels were adapted to black- or white-coloured backgrounds and the pituitary glands were prepared for light and electron microscopy. Immunocytochemical staining was used to study the distribution of the neurohypophysial melanin-concentrating hormone in the neurointermediate lobe. The hormone was located in small, elliptical, electron-opaque neurosecretory granules, measuring approximately 120 x 90 nm. The neurones terminated on blood vessels in the centre of the neurohypophysis and on the basement membrane separating neural and intermediate lobe tissues. The results of both light and electron immunocytochemistry and of radioimmunoassay are consistent with a higher rate of hormone release from eels adapted to white backgrounds than from those adapted to black backgrounds. In addition to this, when fish that had been adapted to white tanks were transferred to black tanks, there was an accumulation of irMCH in the gland and an increased numerical density of secretory granules at nerve terminals. These results reinforce the proposal that MCH is released during adaptation to a white background, to cause melanin concentration and to inhibit MSH release, and that its release is halted in black-adapted fish.

Adaptation, Biological↗
PubMed1987

Ultrastructural demonstration that melanin-concentrating hormone-like and alpha-melanocyte-stimulating hormone-like immunoreactive molecules coexist in the same neurosecretory granules.

Using immunocytochemical methods at the electron microscope level, immunoreactivity for both melanin-concentrating hormone (MCH) and alpha-melanocyte-stimulating hormone (alpha-MSH) has been demonstrated in the carp neurohypophysis. A double-labelling technique, using colloidal gold probes of different sizes showed that immunoreactivity to both molecules coexists within the same neurosecretory granules in some neurones, while in other neurones the granules exhibit only MCH-like immunoreactivity. These observations suggest that the two immunoreactivities are attributable to separate molecules; if they are derived from the same precursor molecule, then this must be cleaved differently in the two sets of neurones. The absence of adrenocorticotropic hormone (ACTH)-like immunostaining in any neurosecretory granule might suggest the alpha-MSH-like molecule is not derived from the conventional pro-opiomelanocortin precursor.

Animals↗
PubMed1984

Single-cell protein.

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Carbohydrate Metabolism↗
PubMed1977

Ribosomal abnormality in recA mutants of Escherichia coli.

The tif-1 mutation has been shown to affect protein synthesis in vitro by increasing translational ambiguity (Ephrati-Elizur, Luther-Davies and Hayes, 1976). It is demonstrated here that some recA mutations confer similar abnormality. By comparing suitable combinations of ribosomes and soluble proteins from recA+ and recA cells the defect is shown to be associated with ribosomes. The recA mutation, which suppresses most phenotype characteristics of the tif-1 mutation (Castellazzi, George and Buttin, 1972(b)) does not suppress the ribosomal abnormality. Since the closely linked tif-1 and recA mutations lead to the expression of a common property they may be in the same gene.

Bacterial Proteins↗
PubMed1975

recA+-dependent inactivation of the lambda repressor in Escherichia coli lysogens by gamma-radiation and by tif expression.

When gamma lysogens of E. coli are induced by gamma-radiation the gamma repressor, as measured by its specific binding to gamma DNA, is rapidly inactivated by a recA+-dependent process which does not require new protein synthesis. This rapid inactivation is similar to inactivation of repressor by expression of the temperature sensitive E. coli mutation tif. In contrast, induction by UV irradiation or mitomycin C treatment requires new protein synthesis and there is a lag before the repressor is inactivated (Tomizawa and Ogawa, 1967; Shinagawa and Itoh, 1973).

Coliphages↗
PubMedDate not supplied

Validation of cine phase-contrast MR imaging for motion analysis.

The accuracy of cine phase-contrast magnetic resonance (MR) imaging for motion analysis was evaluated. By using a rotating phantom and postprocessing algorithm for phase tracking, errors arising during data acquisition were identified and compensation methods were developed. A spatially varying background phase offset in the velocity images was found to be due to eddy current-induced fields. The magnitude of the offset was in the range of 0-20 cm/sec, which is of the same order of magnitude as cardiac contractile velocities. Background offset is thus an important source of error in tracking cardiac motion. Study of different tracking algorithms revealed the need for an integration scheme using motion terms higher than velocity. Also, considerable improvement in the accuracy and stability of the predicted trajectories was obtained by averaging the trajectories proceeding both forward and backward in time from the starting point. With the algorithm developed, the motion of the phantom was tracked through a complete rotation of the phantom to an accuracy of 2 pixels.

Algorithms↗
PubMedDate not supplied

Riboflavin uptake in microvillous and basal membrane vesicles isolated from full-term human placentas.

Riboflavin uptake was characterized using membrane vesicles isolated from the apical (maternal-facing) and basal (fetal-facing) membranes of the syncytiotrophoblast from full-term human placentas. Equilibrium [3H]riboflavin uptake was insensitive to variations in incubation medium osmolarity in contrast to [3H]alanine uptake into an osmotically sensitive space. Osmotic insensitivity suggested riboflavin binding to a membrane component. The dissociation constant of riboflavin binding was similar in microvillous (Kd = 2 microM) and basal membrane vesicles (Kd = 1 microM). Binding capacity was significantly higher in microvillous membranes (Bmax = 11.9 pmol/mg protein). The relatively high affinity binding to the membrane vesicles may represent a first step in riboflavin transport.

Alanine↗