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B C Elford

Publications and source records attributed to B C Elford.

32 records · Page 2Linked to original sources

Selective stage-specific changes in the permeability to small hydrophilic solutes of human erythrocytes infected with Plasmodium falciparum.

The permeability characteristics of Plasmodium falciparum-infected human erythrocytes to various 3H-labelled solutes were measured during the maturation of the parasites in sorbitol-synchronised cultures. Using [14C]inulin as the extracellular marker, estimates were made of the influx kinetics of [3H]amino acids into trichloroacetic acid (TCA)-soluble pools within the erythrocyte and concomitant incorporation into TCA-precipitable material. These measurements provided values of the rates of protein synthesis by the parasite and the initial influx rates for the transport of precursor amino acids into the erythrocyte. For about 12-15 h after parasitisation, the influx of L-[3H]glutamine remained at a low level comparable to that in the uninfected cell (2-9 nmol g-1 cells min-1). As pigment appeared in the trophozoite, the initial rate of influx of L-glutamine increased to a value up to 100-fold higher than in the uninfected erythrocyte. The increase in permeability affected only the parasitised cells in a culture of partially infected erythrocytes, and was selective with respect to substrate since the influx kinetics for both [3H]isoleucine and [3H]arginine were not affected by parasitisation. The permeability changes occurred mainly over a 4-8 h period in the development of the young trophozoite, during which time [3H]glycine influx was enhanced by a factor of 3-10, with a comparable increase in the uptake of myo-[3H]inositol. L-[3H]glutamate, which did not penetrate significantly into uninfected erythrocytes, entered red cells infected with mature trophozoites at a rate which was much less than 1% of the parasite-induced-L-glutamine influx. At the stages when the permeability to L-glutamine was markedly enhanced, parasitised cells remained impermeable to [3H]sucrose. An analysis of the relative 3H activities in glutathione and free amino acid pools indicated that, if L-glutamine permeation did not increase during parasite maturation beyond the ring stage, or was blocked by a potential antimalarial compound, an insufficient supply of L-glutamine would be available for the increased rates of parasite protein synthesis and glutathione turnover within the red cell.

Animals↗

Interactions between temperature and tonicity on cation transport in dog red cells.

1. The temperature-dependence of the uptake of 24Na and 42K into dog red cells between 38 and 4 degrees C has been investigated. The effects on the cation fluxes of partial dehydration of the cells in hyperosmolar sucrose (50-125 mM) have also been studied. 2. A Hamilton gas-tight syringe was used to pipette accurately reproducible volumes of packed cells which contained in addition to 24Na or 42K either [131I]albumin or [51Cr]EDTA as extracellular markers. 3. At 38 degrees C Na flux (m-equiv/l. isosmolar cell volume. hr) increased from 2-8 +/- 0-1 (n = 8) in cells of normal volume to 226 +/- 8 (n = 8) when the cells were shrunken by 27-4 +/- 0-6% (n = 8) in media containing sucrose (100 mM). K influx remained relatively constant under these conditions. 4. The exchange of 24Na in shrunken cells followed a single exponential time course but about 9% of the intracellular Na apparently did not exchange with 24Na in the bathing medium. 5. The steady-state influx of Na in cells of normal volume was maximal at about 22 degrees C. The temperature dependence of the Na fluxes in shrunken cells was described by an Arrhenius relationship with a change in slope at about 22 degrees C. 6. The K influx in cells of normal volume decreased as the temperature was lowered from 38 degrees C, to about 12 degrees C, at which temperature the flux was at a well defined minimum. Above 12 degrees C, cell shrinkage had hardly any effect on K influx, but below 12 degrees C the influx in shrunken cells was significantly less than in cells of normal volume. 7. The selective increase in Na flux induced by cell shrinkage results from a Na:Na exchange process which cannot be explained in terms of Ussing's (1947) model of carrier-mediated exchange diffusion. 8. The lack of coupling between the effects of temperature and cell volume on the fluxes of Na and K indicates that localized structural changes of lipid-protein complexes specific for Na or K are responsible for the cation transport characteristics of dog red cells, and that phase transitions in the lipids of the cell membrane are unlikely to account for the temperature dependence of the fluxes.

Animals↗

Diffusion and distribution of dimethyl sulphoxide in the isolated guinea-pig taenia coli.

1. The diffusion of the cryoprotective non-electrolyte dimethyl sulphoxide (DMSO) in the isolated guinea-pig taenia coli at 37, 25 and 0 degrees C has been studied using [(35)S]DMSO.2. Within 1 hr after immersion at 37 degrees C in Krebs solution containing 20% (w/v) DMSO and trace amounts of [(35)S]DMSO, the non-electrolyte was distributed uniformly throughout a volume equivalent to the total initial water content of the muscle.3. The kinetics of efflux of [(35)S]DMSO from muscles at constant volume were analysed on the basis of two models: one incorporated radial diffusion in extracellular fluid with simultaneous permeation into the cells, the other involved only radial diffusion in homogeneous cylinders of tissue having no internal barriers to diffusion; the former was found to give a better representation of the efflux kinetics.4. If it was assumed that the rate of diffusion of DMSO in the extracellular space of taenia coli was the same as that in the bathing medium, the values of the extracellular space and the permeability of smooth muscle to DMSO, obtained from the analysis of the efflux kinetics, were 454 +/- 19 ml./kg and 2.36 +/- 0.05 x 10(-6) cm sec(-1) at 37 degrees C.5. The activation energy for the transfer of DMSO across the surface of the cell was estimated to be 6.0 kcal/mole at 37 degrees C, 6.6 kcal/mole at 25 degrees C and 11.6 kcal/mole at 0 degrees C, indicating either that the equivalent pore radius of the cells decreased with temperature or that the cell permeability represented the sum of two fluxes, one through the aqueous pores of the cell and the other through the lipid phase of the cell membrane, each with a different energy of activation.6. A net flux of water across the surface of the cells, superimposed on the efflux of DMSO, markedly affected the rate of diffusion of the non-electrolyte out of the whole tissue; however, it was considered that an analysis of the efflux kinetics was not possible under these conditions.7. These results provide a basis for methods which will be used to investigate the possibility of preserving tissue in unfrozen aqueous media at sub-zero temperatures.

Animals↗

Permeation and distribution of deuterated and tritiated water in smooth and striated muscle.

1. The diffusion of deuterated water (HDO) into and out of isolated guinea-pig taenia coli and frog sartorius was followed by means of a recording electronic microbalance.2. A comparison was made of the rates of efflux of deuterated and tritiated (HTO) water using the microbalance and a standard tracer technique.3. The time course of the exchange of HDO in both smooth and striated muscle was fitted to the sum of two exponential functions; however, it was not possible to correlate sizes of the intra- and extracellular spaces in the muscles with the sizes of the two compartments in model systems based on compartmental theory of tracer kinetics.4. The efflux of HDO, which was measured by means of the microbalance under stagnant conditions, occurred at a rate approximately 3 times slower than the efflux of HTO which took place while muscles were agitated vigorously in their bathing media.5. There was a marked deviation at short times from a double exponential time course during the efflux of HTO.6. Values of the tracer water permeability, P(d), of smooth and striated muscle fibres, and the extracellular space, E, of the muscles were obtained using a computer to numerically evaluate the series solutions of the equations describing the permeation of labelled water in the muscles.7. It was necessary to assume a value for the diffusion coefficient of the tracer in extracellular fluid in order to compute a unique fit to an exchange curve.8. Values of P(d) and E given by the analysis depended upon the method used for monitoring the exchange, and an interpretation of this dependence was made difficult by the variation in the thickness of unstirred layers on the surface of the muscles under the different experimental conditions.

Animals↗