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At least 685 records · Page 38Linked to original sources

Visualization of subtle contrast-related intensity changes using temporal correlation.

Contrast-enhanced magnetic resonance imaging (MRI) is a promising method for investigating the breakdown of the blood-retinal barrier (BRB). However, subtle intensity changes due to low concentrations of contrast agent can be difficult to detect without observer bias. In this study, we developed a temporal correlation method for detecting these subtle signal intensity changes. The method was evaluated in eyes with chemically induced retinal lesions of known size. A time series of MRI data were collected following i.v. administration of different doses of gadolinium-diethylaminetriaminepentaacetic acid (0.05, 0.1, 0.5 mmol/kg). These time course images were analyzed by temporal correlation to a reference enhancement curve. The reference curve was generated based on a validated theoretical enhancement curve. The temporal correlation method detected signal intensity changes in cases where the changes were too subtle to be visible on a postinjection image or a subtraction image (obtained by subtracting the precontrast image from the final image in the time course set). In addition, assessment of leakage was performed by viewing each image in the set with an eight gray-level palette. Areas of leakage identified in this manner corresponded to those identified by temporal correlation, a finding which supports the validity of the temporal correlation method. These results suggest that temporal correlation may be a time-efficient way to screen large numbers of image data sets using an objective, user-independent criterion.

Algorithms↗

Microbial transformations of steroids--VIII. Transformation of progesterone by whole cells and microsomes of Aspergillus fumigatus.

The filamentous fungus, Aspergillus fumigatus, efficiently hydroxylated exogenous progesterone producing, after 3 h of incubation, 11 alpha- and 15 beta-hydroxyprogesterone as major products, 7 beta-hydroxyprogesterone as a minor product and trace amounts of 7 beta, 15 beta- and 11 alpha, 15 beta-dihydroxyprogesterone. After 72 h the dihydroxyprogesterones were the sole metabolites in the culture medium. Microsomes, prepared by Ca2+ precipitation, catalysed only monohydroxylation of progesterone at the same sites as whole cells. Hydroxylation was dependent on NADPH (but not NADH) which was replaceable by NaIO4. Hydroxylation was inhibited by carbon monoxide and by the azole fungicide, ketoconazole. Microsomes gave a dithionite-reduced, carbon monoxide difference absorbance spectrum with a peak at 448 nm and a Type-I progesterone-binding spectrum typical of cytochrome P450 interaction with substrate. Ketoconazole inhibition studies suggest the presence of two non-inducible cytochrome P450 progesterone hydroxylases, one possessing 7 beta site-selectivity, the other 11 alpha/15 beta site-selectivity.

Aspergillus fumigatus↗

High-sensitive determination of coenzyme Q(10) in iodinate-beta-cyclodextrin medium by inclusion reaction and catalytic polarography.

A novel polarographic method for the determination of coenzyme Q(10) in beta-cyclodextrin (beta-CD) and iodinate system is proposed. The stability of coenzyme Q(10) to light was improved by the formation of coenzyme Q(10)-beta-CD inclusion complex. In addition, the sensitivity for the determination of coenzyme Q(10) was enhanced by both the formation and the polarographic catalytic wave of the inclusion complex in the presence of iodinate. In 0.1 mol/L HAc-NaAc (pH 4.7)-5.0 x 10(-5) mol/L beta-CD-1.2 x 10(-3) mol/L potassium iodinate-ethanol/water (60:40, v/v) medium, coenzyme Q(10)-beta-CD inclusion complex yielded a sensitive association/parallel catalytic wave. The second-order derivative peak current of the catalytic wave was proportional to coenzyme Q(10) concentration in the range of 6.0 x 10(-8)-2.5 x 10(-7) mol/L, and the detection limit was 1.0 x 10(-8) mol/L. The proposed method has high analytical sensitivity and is allowed to determine coenzyme Q(10) under light.

Catalysis↗

Monte Carlo simulation of gamma-ray spectra from natural radionuclides recorded by a NaI detector in the marine environment.

The GEANT4 Monte Carlo code has been used to simulate gamma-ray spectra of natural radionuclides collected by a NaI scintillation detector immersed in seawater. The gamma-rays emitted from the decay of (40)K, and the series of (232)Th and (238)U, were used to describe the radioactive water source around the NaI crystal. The simulated gamma-ray spectra were compared with real data recorded in situ by a newly constructed NaI spectrometer and were found to be in good agreement. The NaI spectrometer was calibrated in the laboratory in a water tank, before its deployment in seawater. Activity concentrations were deduced from the gamma-ray spectra and discussed in comparison with results from the literature.

Computer Simulation↗

Reactive liposomes encapsulating a glucose oxidase-peroxidase system with antibacterial activity.

Liposomes were prepared from phospholipid mixtures of dipalmitoylphosphatidylcholine (DPPC) and phosphatidylinositol (PI), encapsulating the enzymes glucose oxidase (GO) and GO in combination with horse radish peroxidase (HRP) by both extrusion (VET) and reverse-phase evaporation (REV). The optimum level of PI in DPPC/PI liposomes for targeting to biofilms of the oral bacterium Streptococcus gordonii has been established. The liposomes were characterised in terms of the content and activity of the encapsulated enzymes. The antibacterial activity of these 'reactive' liposomes arising from hydrogen peroxide and oxyacids in the presence of the substrates glucose and iodide ions, after targeting to the biofilms, were measured both as a function of liposome-biofilm incubation time and incubation time with the substrates. Bacterial inhibition increases with both liposome-biofilm and substrate-biofilm incubation time and with the extent of enzyme encapsulation. The reactive liposomes also display antibacterial activity in the presence of saliva. The reactive liposomes have potential value in the context of oral hygiene.

Anti-Bacterial Agents↗

Factors affecting the sequestration of aflatoxin by Lactobacillus rhamnosus strain GG.

The interaction of a potent carcinogen, aflatoxin B(1) (AFB(1)), with a probiotic strain of lactic acid bacteria, Lactobacillus rhamnosus strain GG (GG), has been investigated. The binding of AFB(1) to GG in the late exponential-early stationary phase was studied for viable, heat-killed and acid-killed bacteria. In general, viable, heat-killed and acid-killed GG responded in a similar manner. The effects of pronase E, lipase and m-periodate on AFB(1) binding and release were consistent with AFB(1) binding predominantly to carbohydrate components of the bacteria. The effect of urea suggested hydrophobic interactions play a major role in binding. Increasing concentration (0.01-1 M) of NaCl or CaCl(2) had minor effects on AFB(1) binding suggesting some involvement of electrostatic interactions. An increase in pH from 2.5 to 8.5 had no effect on AFB(1) binding but decreased binding of AFB(2a), possibly due to hydrogen bonding interactions.

Aflatoxin B1↗

Reorganization of actin microfilaments and microtubules in regenerating retinal pigment epithelium.

Retinal pigment epithelium (RPE) regenerating after experimental damage in rabbits exhibits major changes in cell shape, polarity and junctions--features that depend on the cytoskeleton. This report correlates these changes with the redistribution of actin microfilaments and microtubules, using electron microscopy and confocal laser scanning microscopy. We compare immature cells with the more mature cells that form the new epithelial monolayer. Two populations of immature RPE cells are interspersed at the edge of the regenerating RPE sheet. One population of immature cells makes few junctions with their neighbors or the basement membrane. They form pseudopodia and exhibit a prominent network of actin microfilaments beneath the plasma membrane. These cells are probably motile and advance the epithelial sheet. Another population of immature cells contains numerous stress fibers that insert into large basement membrane attachments. The cells make focal adhesions with their neighbors, rather than the junctional complexes characteristic of mature RPE cells. These cells are probably not motile and mature into the cells forming the new monolayer--cuboidal cells with numerous basal folds and apical villi and a complete belt of intercellular junctions. Stress fibers are lost as the circumferential bundle associated with the zonula adherens re-forms. Microtubules, which form prominent longitudinal bundles running through the processes of immature cells, take on the meshwork organization characteristic of mature RPE as the immature cells differentiate.

Actin Cytoskeleton↗

Local pressure effects on vitreous kinetics.

Fluorescein (F) or carboxyfluorescein (CF) was injected subconjunctivally in rabbits and its penetration into and subsequent loss from the vitreous body and anterior segment was measured. Pressure was applied over the site of the injected dye, sufficient to close down the local choroidal circulation. This raised the penetration of F about 30 times and delayed its loss, and raised the penetration of CF about seven times without affecting its loss rate. Cooling the tissue, in addition to local pressure, had little additional effect on the penetration. When F was made to penetrate by transcleral iontophoresis, pressure over the site had no effect on its kinetics. This behavior was compared with the predictions by a mathematical model in which the dye was released as a pulse at the inside of the retina and then diffused freely in the vitreous, loss occurring at the retinal surface by an outward transport process. Two experimental observations, the time at which the fluorescence behind the lens reached its peak and its subsequent loss rate, were related in a graphical representation to two physiological parameters, the rate of diffusion of the dye in the vitreous body and the outward permeability of the retinal layer. In some sets of experiments, particularly those in which iontophoresis was used, the kinetics conformed well to the predictions of this model. In other cases, the agreement was poor, and mechanisms such as storage of dye in the retina and its penetration into the vitreous by an anterior pathway appear to play a role.

Animals↗

The effect of anions on azide binding to myoglobin: an unusual functional modulation.

The effect of increasing concentrations of several anions on the azide (N(-)(3)) binding properties of sperm whale and horse ferric myoglobin has been studied. Surprisingly, a number of anions may act as heterotropic effectors, decreasing the affinity of myoglobins for N(-)(3), in the following order: ClO(-)(4)=I(-)>Br(-)>Cl(-) and SO(2-)(4), which mirrors the increase in their charge density. The largest effects were measured using ClO(-)(4) and I(-), which produce a 4-fold and 8-fold reduction of the N(-)(3) binding affinity in horse and sperm whale myoglobins, respectively. A dissociation equilibrium constant (K(d)) ranging from 150 to 250 mM was estimated for ClO(-)(4) and I(-) binding to myoglobins. In order to analyse the molecular mechanism producing the reduction of the N(-)(3) binding affinity to ferric myoglobin, the potential anionic binding sites within ferric myoglobin were investigated by a molecular modelling study using the program Grid. Analysis of the theoretical results suggests two particularly favourable binding sites: the first, next to the distal side of the haem, whose occupancy might alter the electrostatic potential surrounding the bound N(-)(3); the second, involving residues of helices B and G which are far from the haem iron atom, thus implying a long range effect on the bound N(-)(3). Based on the evidence that no significant conformational changes are found in the three-dimensional structures of N(-)(3)-free and N(-)(3)-bound myoglobin and on previous results on N(-)(3) binding to ferric myoglobin mutants in CD3 positions, we favour the first hypothesis, suggesting that the functional heterotropic modulation of monomeric myoglobin is mainly depending on a decrease of the positive charge density induced by the binding of anions to the haem distal side.

Allosteric Regulation↗

Capillary zone electrophoresis and ion-exchange capillary electrochromatography: analytical tools for probing the Hanford nuclear site environment.

Ion-exchange capillary electrochromatography (IE-CEC) is a relatively new separation technique based on the combination of ion-exchange chromatographic and electrophoretic separation mechanisms. IE-CEC offers both the efficiency of capillary electrophoresis and the selectivity and sample capacity of ion-exchange chromatography. The utility of the method was examined with I- and IO3-, which are common constituents of nuclear wastes at Hanford, Washington and other U.S. Department of Energy (DoE) sites, and ReO4-, a surrogate for TcO4-. The advantages and limitations of IE-CEC relative to capillary zone electrophoresis (CZE) are explored. The chief advantages are increased loading capacity and an alternative selectivity to that of CZE, in addition to increased efficiency (relative to conventional ion-exchange chromatography). The run-to-run reproducibility of IE-CEC, however, was found to be a limitation of the technique.

Chromatography, Ion Exchange↗

Determination of some aminobenzoic acid derivatives: glafenine and metoclopramide.

Simple, sensitive and accurate spectrophotometric methods for the determination of glafenine and metoclopramide hydrochloride are described. The first method is based on the oxidation of glafenine with iodic acid in strong acid medium to give a coloured diphenylbenzidine derivative and subsequent measurement of the coloured product at 509 nm. Beer's law is obeyed over the concentration range 2.5-20 microg ml(-1). The second method depends on the interaction of metoclopramide hydrochloride with p-dimethylaminocinnamaldehyde, to give a red coloured schiffs base with an absorbance maximum at 548 nm. Obedience to Beer's law is achieved over the concentration range 5-30 microg ml(-1). First-derivative method is used to overcome the slight interference of p-dimethylaminocinnamaldehyde reagent blank at the wavelength of measurement. Linearity between the peak heights at 576 nm versus concentration range 5-25 microg ml(-1) metoclopromide hydrochloride is obtained. The proposed methods have been successfully applied to the determination of these drugs in commercial products without interference. The validity of the methods is assessed by applying the standard addition technique, the relative standard deviation is less than 1%. The proposed methods are compared with reference methods with good agreement.

Analgesics, Non-Narcotic↗

Spectrophotometry assay of imipramine and desipramine using ammonium metavanadate and its application to pharmaceutical preparations.

Simple and sensitive method for determination of imipramine and desipramine is reported. The procedure is based on the oxidation of the drugs by ammonium metavanadate. Linear calibration graphs were obtained in the concentration range 0.6-40 microg ml(-1) of imipramine and 0.7-35 microg ml(-1) of desipramine with a relative standard deviation (RSD) less than 0.5%. The method was applied to the determination of the drugs in pharmaceutical preparations and compared favourably with independent official methods.

Antidepressive Agents, Tricyclic↗