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Biomedical subjects

T Larsson

Publications and source records attributed to T Larsson.

50 records · Page 3Linked to original sources

Entrapping of the spin label tempocholine into human erythrocytes by resealing after hyposmolar stress. Comparison with haemolysis. The effects of some membrane-active substances.

Human erythrocytes were subjected to a sudden hyposmolar stress by suspension in solutions of varying salt concentrations in the presence of the spin label tempocholine. The enlarged pores in the erythrocyte membranes produced by the influx of water, followed by stretching, allowed the passage of the spin label, so that a certain amount of tempocholine was entrapped when the erythrocytes spontaneously resealed with closing of the pores. The excess of spin label in the external solution was then reduced to a diamagnetic species by the addition of ascorbic acid. The positively charged tempocholine and the ascorbic acid did not penetrate properly resealed erythrocytes, so that the electron spin resonance (ESR) signal from the entrapped spin label constituted a measure of the effective resealing of the pores and rifts in the membrane subsequent to hyposmolar stress. Some drug substances were found to influence the entrapping curves obtained when the amount of entrapped spin label was plotted against the osmolarity. Chlorpromazine, trifluoperazine, nicardipine, amperozide and haloperidol gave rise to a dose-dependent decrease of the entrapping of tempocholine, especially at low osmolarities. The exclusion of Ca2+ and Mg2+ ions from the solutions increased the action of chlorpromazine. The protective action against haemolysis brought about by a number of membrane-active substances at low concentrations [2] had its counterpart in the entrapping curve observed with chlorpromazine at 0.1 mM. It is suggested that the substances in this series exerted their action on the resealing process by interaction with the calmodulin system.

Calcium↗

The efflux of spin label entrapped in human erythrocyte ghosts when suspended in hyposmolar solutions. The effect of chlorpromazine, trifluoperazine, nicardipine and some other membrane active substances.

Human erythrocyte ghosts were loaded with the spin label tempocholine . Once entrapped in the ghosts, this spin label, carrying a positive charge, is not able to penetrate through intact ghost membranes. The ghosts were loaded with spin label to a concentration high enough to introduce exchange broadening of the electron spin resonance (ESR) signal with a relatively small signal amplitude. The efflux of the spin label brought about by hyposmolar stress was studied. The appearance of the label in the relatively large external volume gave rise to an increase of the ESR signal amplitude since the concentration of the spin label outside the ghosts was in the range in which exchange broadening can be excluded. The duration of the efflux following hyposmolar stress was less than half a minute. After this time, the ghosts resealed spontaneously and without restoration of the normal osmolarity. A number of membrane active substances were studied for possible influence on the efflux of spin label induced by hyposmolar stress. The drug substances chlorpromazine, trifluoperazine and nicardipine were found to increase the hyposmolar efflux of spin label. It was suggested that these substances, classified as calcium-antagonists and inhibitors of the calmodulin system, exert their action on the efflux of spin label by interaction with membrane proteins which maintain shape and tension of the erythrocytes.

Calcium Channel Blockers↗

Comparative studies of the binding of some ligands to human serum albumin non-covalently attached to immobilized Cibacron Blue, or covalently immobilized on Sepharose, by column affinity chromatography.

A comparative study of the ligand-binding properties of human serum albumin was performed by the technique of affinity chromatography with the protein attached to immobilized Cibacron Blue F3GA (Blue Sepharose), or covalently immobilized on Sepharose. The binding strength of octanoate, decanoate and dodecanoate is much weaker when human serum albumin is attached to immobilized Cibacron Blue, indicating that the binding sites for fatty acids are involved in the attachment of human serum albumin to immobilized Cibacron Blue. The results revealed additional alterations of the ligand binding when human serum albumin was attached to immobilized Cibacron Blue, involving sites outside of the binding domains of fatty acids. Thus the stereoselective binding of L-tryptophan was abolished, and the resolution of the warfarin enantiomers was impaired. However, the binding strength of warfarin and salicylic acid was rather close to the values observed with human serum albumin covalently immobilized on Sepharose. It is suggested that the availability of the binding sites for L-tryptophan, warfarin and salicylic acid is partially blocked by the complex between albumin and the dye without direct participation in the complex-formation. An alternative interpretation involves an allosteric mechanism brought about by complex-formation between serum albumin and the immobilized Cibacron Blue.

Anthracenes↗

Synthesis and binding to tubulin of an allocolchicine spin probe.

A spin probe for the colchicine binding site on tubulin has been synthesized from allocolchicine. The probe is competitive to colchicine with an apparent inhibition constant of 11 microM while allocolchicine has an inhibition constant of 2 microM. Microtubule assembly is inhibited to 50% by 7 microM of the spin probe. As a mitotic poison the spin probe is less potent than colchicine. These data suggest that the probe binds to the same site on tubulin as colchicine and in spite of the somewhat lower efficiency, it seems to be a valuable tool for the study of the microtubule system.

Animals↗

Quantitative studies on competitive ligand binding to bovine serum albumin by use of the spin label 5-doxyl dodecanoic acid.

The binding of the spin label 5-doxyl dodecanoic acid to bovine serum albumin in phosphate buffer at pH 7.4 was studied by electron spin resonance spectroscopy. Free label and label bound to serum albumin could be quantitatively measured and evaluated from the superposition spectra of these two species with no previous separation. The efficiency relative to the spin label as competitors for binding to serum albumin was studied with salicylic acid and some fatty acids of medium length. The results were represented both by the stoichiometric model involving equilibrium constants Ki, by binding isotherms constructed from the Ki values, and by a purely graphical representation of the experimental data points without connection with any special binding model.

Binding, Competitive↗

The value of thermography and the determination of fibrin-fibrinogen degradation products in the diagnosis of deep venous thrombosis.

Fifty-one patients with leg symptoms indicating deep venous thrombosis (DVT) were investigated concomitantly with thermography and phlebography. Altogether 26 legs with phlebographically proven DVT exhibited a thermographic picture typical of DVT. So, however, did 3 out of 25 legs with no DVT. A 94 percent agreement was found between phlebography and thermography when applied to legs with suspect DVT. The serum levels of fibrin-fibrinogen degradation products (FDP) were assayed in 58 patients attending hospital for suspect DVT. These patients were free from other diseases known to be followed by raised FDP levels. With borderline significance, the DVT group had higher FDP values. The usefulness of this laboratory test in the diagnosis of DVT is, however, questioned.

Adult↗

Identification of two-dimensional gel electrophoresis resolved yeast proteins by matrix-assisted laser desorption ionization mass spectrometry.

Protein extract from yeast cells growing exponentially in saline medium was separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), with the separation in the first dimension on a wide range immobilized pH (3-10) gradient. From one preparative 2-D gel a number of previously identified proteins were used as test material for our initial matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) efforts on large scale rapid protein spot identification. Sample preparation via in-gel trypsin digestion was slightly modified to be compatible to MS analysis, and via this modified procedure MS generated peptide mass profiles could, in most cases with good precision, identify the protein in question. Preferential ionization was tested on a yeast aldehyde dehydrogenase (ALD7), and it was shown that the ionization of some peptides was clearly suppressed by the presence of others. Roughly 50% of the observed peptide masses was found by the search routines in the database, and the mass measurement accuracy of the peptides was within 0.5 Da. Silver-stained gels could be used with good results for the generation of peptides to be analyzed by MALDI-MS. For one of the 2-D resolved proteins, glycerol 3-phosphatase (GPP1), the post-source decay (PSD) spectrum proved crucial in identification.

Amino Acid Sequence↗

Absorbed power distributions from two tilted waveguide applicators.

One major problem in microwave-induced clinical hyperthermia treatment of superficial tumours is to obtain therapeutic temperatures at the tumour periphery and adequate deep heating when using a single applicator. The use of multiple applicators has therefore been investigated in order to improve the power distribution. Anatomical surface topography often permits the application of two tilted applicators, e.g. in the head and neck area, on extremities and on large protruding tumours. Theoretical calculations of the absorbed power distribution from such an applicator configuration were performed in a homogeneous muscle equivalent medium. The power distribution from two conventional radiative apertures (TE10) was studied at different frequencies, aperture sizes, tilting angles and non-coherent or coherent fields in phase, both theoretically and with phantom experiments using a thermographic camera. With controlled phase relations between two tilted applicators excited at the lower microwave or upper radiofrequency band, the absorbed power distribution can be varied in a wide range. The theoretical calculations and thermographic phantom experiments in simple geometries give valuable information on absorbed power distributions and guidance for the location of temperature probes in clinical hyperthermia.

Hyperthermia, Induced↗