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Biomedical subjects

A E Robinson

Publications and source records attributed to A E Robinson.

At least 55 records · Page 3Linked to original sources

Radionuclide determination of right- and left-ventricular mixing.

Previous models for the dynamics of the transit of the bolus of activity of labeled red cells through the chambers of the heart have assumed the complete and instantaneous mixing of tracer with the cardiac blood pool. This assumption can be conveniently tested with a radionuclide procedure. We present a model of the bolus transit through the right and left ventricle that allows the quantitation of the mixing of labeled cells with the residual blood of the cardiac chambers. This model was used to analyze the time-activity curves obtained from a variety of patients undergoing first-pass dynamic scans of the heart. It is shown that technetium-labeled red blood cells and technetium pertechnetate have indistinguishable mixing properties, and the blood in the left ventricle is more thoroughly mixed than that in the right ventricle.

Cardiac Output↗

Cross-linking of bacteriorhodopsin using specific carboxyl modifications and proteolytic cleavage.

Specific carboxyl modification of purple membrane using a water-soluble carbodiimide yielded a mixture of oligomers, revealed by gel electrophoresis. Purple membrane pre-treated with papain or trypsin, cleaving the C-terminal tail, showed the same pattern of cross-linked products. Chymotryptic cleavage released amino acids 1-72 (7kD fragment) from the cross-linked products, as it did with native membrane. The tail and helices A and B are not, therefore, involved in carbodiimide-promoted cross-linking. Similar cleavage of a hydrophobic dihydroquinoline-modified sample showed that mainly intra-molecular cross-linking occurs, with little cross-linking between the large and small chymotryptic fragments.

Bacteriorhodopsins↗

The role of Na2S in anoxygenic photosynthesis and H2 production in the cyanobacterium Nostoc muscorum.

Na2S is known to support anoxygenic photosynthesis in some strains of cyanobacteria and to stimulate H2 production in N2 fixing filaments of Nostoc muscorum. We have shown electron transfer between Na2S and Photosystem I to be dependent on cytochrome b559 which was detected only in vegetative cells. An electron mediator was required to support Na2S driven nitrogenase activity in isolated heterocysts. Na2S was also found to deplete the ATP pool, probably by inhibiting electron transfer from Photosystem I.

Anaerobiosis↗

Effect of specific carboxyl modifications on the blue acid species and O650 photocycle intermediate of bacteriorhodopsin.

Carboxyl groups were specifically modified in purple membranes using hydrophobic or hydrophilic activating reagents and endogenous or exogenous nucleophiles. Modification of surface residues using a negatively charged nucleophile had no effect on blue-species formation, whereas surface or internal carboxyl modification using an uncharged nucleophile prevented blue-species formation. The carboxyl-modified samples showed poor correlation between blue-species and O650 intermediate. These results indicate that formation of the blue-species requires negative surface charge and is not associated with the O650 intermediate of bacteriorhodopsin, as had been previously postulated.

Bacteriorhodopsins↗

Low-temperature magnetic-circular-dichroism spectroscopy of the iron-molybdenum cofactor and the complementary cofactor-less MoFe protein of Klebsiella pneumoniae nitrogenase.

The major metal clusters of the MoFe protein, Kpl , of Klebsiella pneumoniae nitrogenase were characterized separately by low-temperature magnetic-circular-dichroism spectroscopy. The spectra and magnetization curves of the extracted iron-molybdenum cofactor, FeMoco , and of 'P' clusters in NifB - Kpl , the inactive, FeMoco -less, MoFo protein from an nifB mutant, were measured and compared with those of the holoprotein. (When FeMoco and NifB - Kpl are combined, active Kpl is formed.) Reduced NifB - Kpl had a spectrum with a weak, paramagnetic, component superimposed on a diamagnetic background. The paramagnetic component was assigned to a contaminating, e.p.r.-active, species. Thionine-oxidized NifB - Kpl had a spectrum and magnetization properties very similar to those of thionine-oxidized Kpl , demonstrating that the 'P' clusters are not significantly affected by the absence of the FeMoco clusters. The spectra of reduced isolated FeMoco had similar magnetization curves but sharper features and higher intensities than those of this centre in dithionite-reduced Kpl . Furthermore, a shoulder near 580 nm in the Kpl spectrum was absent from that of FeMoco . This may be due to the loss of a ligand or to a change in symmetry of the FeMoco cluster on extraction.

Circular Dichroism↗

Characterization of the Fe-S cluster in aconitase using low temperature magnetic circular dichroism spectroscopy.

Beef heart aconitase has been studied by low temperature magnetic circular dichroism (MCD) spectroscopy in the wavelength region 300 to 1900 nm. Together with parallel electron paramagnetic resonance and activity measurements, these data enable correlations between Fe-S cluster-type and enzymic activity in aconitase. In samples not exposed to extraneous Fe, the Fe-S cluster in aconitase exhibits the characteristic properties of a 3Fe center in both the as isolated and dithionite-reduced states. On the basis of the detailed form of the low temperature MCD spectra, three types of 3Fe center can be distinguished in biological samples. These are typified by the 3Fe centers in aconitase, Desulfovibrio gigas FdII, and Azotobacter crooccocum Fd. In aconitase, maximal enzymic activity is found to be associated with the build-up of [4Fe-4S]2+ clusters in good agreement with the Mössbauer studies of Kent et al. (Kent, T. A., Dreyer, J. L., Kennedy, M. C., Huynh, B. H., Emptage, M. H., Beinert, H., and Münck, E. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 1096-1100). However, significant catalytic activity (approximately 60%) was obtained by reduction of the 3Fe center with dithionite in the absence of added Fe. The form and intensity of the resultant MCD spectrum are consistent with the majority of the Fe being in the form of reduced 3Fe clusters. The possibility that a reduced 3Fe cluster is capable of promoting partial catalytic activity in aconitase is discussed in light of these results.

Aconitate Hydratase↗

Radionuclide determination of cardiac outputs and indices.

A radionuclide procedure for the determination of cardiac output is presented. Cardiac outputs estimated by the proposed method are compared to those obtained by the temperature dilution method during right heart catheterization. A new index of cardiac performance is also presented: cardiac output as a percentage of total blood volume. This index is determined by a totally external count-based procedure.

Adult↗

Sonographic determination of renal volumes in normal neonates.

Renal diseases that affect renal size without altering renal architecture require a quantitative means of detection. A prospective study was undertaken to establish normal values for renal volumes in healthy neonates using sonography. Volumes were determined by two methods; (1) the serial area-volume method using parallel transverse images; and (2) the prolate ellipsoid model of the kidney using orthogonal diameters taken from ultrasound images. Renal volumes for both the right and left kidneys in both sexes were found to be approximately 10 ml. There was no significant difference between the results obtained by either method, nor were there significant differences between the volumes of the right and left kidneys within either sex. No difference in renal volume was noted between sexes. The mean greatest renal length was also computed for right and left kidneys in both sexes. Knowledge of normal renal volumes may aid in the diagnosis of urinary system disorders in neonates.

Female↗

Pilot study for large-scale plasma procurement using automated plasmapheresis.

A pilot study for large-scale automated plasmapheresis using the Haemonetics Model 50 machine was undertaken in the Yorkshire Region of the United Kingdom to determine the viability of such a programme for national self-sufficiency in fresh plasma procurement for factor VIII concentrate production. The study was designed to resolve three areas of concern: donor safety and recruitment; a cost analysis, and the choice of anticoagulant for optimum factor VIII yields. The results show that large-scale automated plasmapheresis could safely and economically produce high-quality source plasma necessary for national self-sufficiency.

Anticoagulants↗

Abrupt termination of the common bile duct: a sign of malignancy identified by high-resolution real-time sonography.

Although contrast cholangiography has a greater accuracy than sonography in determining the precise nature of extrahepatic biliary obstruction, it is not without risk. High-resolution real-time sonography is frequently adequate to evaluate the extrahepatic biliary system in multiple planes without known risk to the patient. In a one-year period the authors prospectively observed eight instances of distal biliary obstruction with abrupt termination of the common bile duct, and all were associated with malignant obstruction. There was no instance of abrupt ductal termination secondary to benign disease in this series. Malignancy should be suspected when careful sonograms along the longitudinal axis of an obstructed distal common bile duct reveal an abrupt termination.

Aged↗

Determination of left-ventricular volume from first-pass kinetics of labeled red cells.

A mathematical model is presented for the dynamics of a bolus of technetium-99m-labeled red blood cells through the left ventricle. It is used to correct for attenuation the count rate observed over the left ventricle during a conventional gated blood-pool study. The left-ventricular volumes are calculated from the corrected count rates and expressed as a percentage of total blood volume, or in absolute terms if a blood sample is obtained. The procedure is applied to a number of patients with nonvalvular cardiac disease. Cardiac volumes determined by the method are found to correlate well (r = 0.98) with those determined by contrast left ventriculography. The method is simple, requires no special equipment, and can be applied with existing computer software.

Adult↗

The three-iron cluster in a ferredoxin from Desulphovibrio gigas. A low-temperature magnetic circular dichroism study.

Ferredoxin II from Desulphovibrio gigas is a tetrameric protein containing a novel iron-sulphur cluster consisting of three iron atoms. The low-temperature magnetic circular dichroism (MCD) spectra of the oxidized and dithionite-reduced forms of ferredoxin II have been measured over the wavelength range approx. 300-800 nm. Both oxidation levels of the cluster are shown to be paramagnetic, although only the oxidized form gives an EPR signal. MCD magnetization curves have been constructed over the temperature range approx. 1.5-150 K and at fields between 0 and 5.1 Tesla. The curve for the oxidized protein can be fitted to a ground state of spin S = 1/2 with an isotropic g factor of 2.01. There is evidence for the thermal population of a low-lying electronic state above 50 K. The reduced protein gives a distinctive set of magnetization curves that are tentatively assigned to a ground state of S = 2, with a predominantly axial zero-field distortion that leaves the doublet Ms = +/-2 lowest in energy. The zero-field components have a maximum energy spread of approx. 15 cm-1. which places an upper limit of 4 cm-1 on the axial zero-field parameter D. The MCD spectra of the oxidized and reduced forms of the cluster are quite distinctive from one another. The spectra of the oxidized state are also different from those of oxidized high-potential iron protein from Chromatium and should provide a useful criterion for distinguishing between four- and three-iron clusters in their highest oxidation levels.

Circular Dichroism↗

Low-temperature magnetic circular dichroism spectra and magnetisation curves of 4Fe clusters in iron-sulphur proteins from Chromatium and Clostridium pasteurianum.

The magnetic circular dichroism (MCD) spectra of the 4Fe clusters in the iron-sulphur proteins high-potential iron protein from Chromatium and the 8Fe ferredoxin from Clostridium pasteurianum have been measured over the wavelength range 300-800 nm at temperatures between approx. 1.5 and 50 K and at magnetic fields up to 5 tesla. In both cases the proteins have been studied in the oxidized and reduced states. The reduced state of high-potential iron protein gives a temperature-independent MCD spectrum up to 20 K, confirming the diamagetism of this state at low temperature. The MCD spectrum of samples of oxidized ferredoxin invariably show the presence of a low concentration of a paramagnetic species, in agreement with the observation that the EPR spectrum always shows a signal at g = 2.01. The paramagnetic MCD spectrum runs across the whole of the wavelength range studied and therefore most probably originates from an iron-sulphur centre. The diamagnetic component of the MCD spectrum of oxidized ferredoxin is very similar to that of reduced high-potential iron protein. The low-temperature MCD spectra of oxidized high-potential iron protein and reduced ferredoxin reveal intense, temperature-dependent bands. The spectra are highly structured with that of high-potential iron protein showing a large number of electronic transitions across the visible region. The MCD spectra of the two different oxidation levels are quite distinctive and should provide a means of establishing the identity of these state of 4Fe clusters in more complex proteins. MCD magnetisation curves have been constructed from detailed studies of the field and temperature dependence of the MCD spectra of the two paramagnetic oxidation states. These plots can be satisfactorily fitted to the theoretically computed curves for an S = 1/2 ground state with the g factors experimentally determined by EPR spectroscopy. The low-temperature MCD spectra of the reduced 2Fe-2S ferredoxin from Spirulina maxima are also presented and MCD magnetisation curves plotted and fitted to the experimentally determined g factors.

Chromatium↗