Biomedical subjects
Publications and source records attributed to L Villegas.
Cassava (Manihot esculenta Crantz) protoplast was analyzed by using isoelectric focusing techniques. Two populations, representing 68 and 32% of the total sample, with mean isoelectric points of 4.48 and 4.60, were obtained using mesophyll protoplasts. The use of this technique allows demonstration of a discontinuous distribution of protoplast isoelectric point from one species according to their surface potential.
Serosal-to-mucosal and mucosal-to-serosal diffusion of 14C-labelled inulin, sucrose, erythritol and propionamide was compared with 3HHO diffusion in mucosae incubated with isosmotic solutions at both surfaces, as well as isosmotic solution at serosal surface and hyperosmotic solution at the mucosal surface. The use of a hyperosmotic solution at the mucosal surface significantly increases unidirectional diffusion fluxes of inulin and of sucrose. To a nonsignificant extent, it affects the fluxes of erythritol and propionamide and significantly reduces the 3HHO diffusion. A size increment of the diffusion path utilized by the larger molecules is proposed.
The asymmetry of the osmotic response of the frog gastric mucosa has been further analyzed by studying the effect of external tonicity changes on the water diffusion fluxes. Hyperosmotic solution at the serosal surface does not affect the water diffusion fluxes. Hyperosmotic solution at the mucosal surface, with isosmotic solution at the serosal surface, significantly reduces (P less than 0.001) the serosal-to-mucosal and the mucosal-to-serosal water diffusion. An increment in the restriction offered by the mucosa to water diffusion by effect of hypertonicity at the mucosal surface is proposed.
The effects of hyperosmolality of the serosal and mucosal bathing solutions on the transmucosal net water and ion fluxes were studied in frog gastric mucosa. Addition of 100 mosM glucose to the solution at the serosal surface produces a reversed mucosal-to-serosal net water flux of 7.1 +/- 1.4 microliter . cm-2 . h-1. When added to the abolished spontaneous net water flux, this results in an increment in the net water flux of -17.8 +/- 1.4 microliter . cm-2 . h-1. Addition of the same amount of glucose to the solution at the mucosal surface produces an increment in the serosal-to-mucosal net water flux of 3.7 +/- 1.1 microliter . cm-2 . h-1 when the solution at the opposite surface was kept in 220 mosmol/kg H2O. Simultaneous increments of both solutions of 320 to 420 and 420 to 520 mosM changes the osmotic serosal-to-mucosal induced fluxes to 1.9 +/- 0.9 and 3.4 +/- 1.6 microliter . cm-2 . h-1, respectively. The initial spontaneous net water fluxes measured in 220, 320, and 420 mosM solutions were 11.3 +/- 0.9, 6.9 +/- 1.6, and -1.5 +/- 1.5 microliter . cm-2 . h-1. It is proposed that the osmotic water flux is asymmetric, independent of the solutions tonicities, and not significantly affected by the sweep of solutes at the mucosal surface.
Intracellular analyses of the electrical potential response to current pulses were performed in epithelial and oxyntic cells of the frog gastric mucosa. Independently of the use of Cl- or SO(2-)4 solutions and of the transmucosal potential difference measured, the resistance and slow potential changes observed across the mucosal surface of epithelial cells are not different from those registered across the serosal surface. In the oxyntic cells the resistance and slow potential changes are significantly higher across the mucosal than across the serosal surface, when the solution in contact with the serosal surface is positive with respect to the solution in contact with the mucosal surface. These differences were not observed when the transmucosal potential difference was reversed. It is proposed that the ionic concentration changes, by effect of current flow across the mucosa, occur in the oxyntic cells and that these changes are the origin of the slow transmucosal potential change. The epithelial cells act as a passive shunt.
The effects of hydrostatic pressure differences up to 0.4 atm/413 cmH2O were studied on frog gastric mucosa in vitro. Net water flux, transmucosal electrical potential difference, and acid secretion were measured. A significant correlation between hydrostatistic pressure difference and net water flow (r=0.77) was obtained. The intercept of the regression line, at zero hydrostatic pressure difference, is 9.3 +/- 0.5 microliter/cm2.h, and the slope 42.9 +/- 3.2 microliter/cm2.atm.h. No significant correlation was obtained between the hydrostatic pressure difference and the transmucosal potential difference (P greater than 0.20), the acid secretion (P greater than 0.20), or the nonacidic chloride transport, measured as short-circuit current (P greater than 0.20). Hydrostatic water flux is compared to osmotically induced flux previously reported. It is proposed that the difference between hydrostatic and osmotic induced water fluxes is due to the area of cells exposed to the pressures. Only part of surface cells are directly exposed to the osmotic pressure due to the presence of restricted extracellular compartments.
The dimethyloxazolidine dione distribution in the extracellular compartments of the frog gastric mucosa was analyzed by washout kinetics. The volumes of the two extracellular compartments, serosal and mucosal, were estimated by inulin washout as 0.435 +/- 0.019 and 0.176 +/- 0.018 microliter/microliter tissue water, respectively. In the serosal extracellular space, significant dimethyloxazolidine dione accumulations of 2.63 +/- 0.25, 2.28 +/- 0.16, and 1.86 +/- 0.08 times that of the bathing media were found for bathing solutions with pH values of 6.9, 7.4, and 7.9 respectively. A high pH of the serosal extracellular fluid by itself could not account for the high values of dimethyloxazolidine dione accumulation. A difference in the total dimethyloxazolidine dione accumulation requires: (a) the existence of differences in the pH values and also the existence of a difference in the diffusion coefficient of the two forms of dimethyloxazolidine dione; or (b), a binding of one of the two forms, i.e., binding of dimethyloxazolidine dione form by fixed charges.
The effects of symmetric changes of the mucosal and serosal bathing solution on cell water content, net ion flux, and net water movement were studied in the isolated frog gastric mucosa. Similar to transmucosal concentration gradients that induce water movement and changes in cell water content, symmetric osmolality changes of the bathing solutions also produce changes in these functional parameters. Thus, increments from 165 to 286 mosmol/kg water in the osmolality of both solutions reduce cell water content from 2.37 plus or minus 0.12 to 1.30 plus or minus 0.20 ml/g wt, the net ion flux (acid secretion plus short-circuit current) from 4.83 plus or minus 0.36 to 3.44 plus or minus 0.26 mueq/cm2 per h, and the net water flux from 10.6 plus or minus 1.1 to 2.4 plus or minus 1.2 mul/cm2 per h. These osmotically induced flux changes of water and ions must be considered when osmotic gradients are being used to generate and to evaluate water movement across the gastric mucosa.
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Sodium, potassium, and chloride concentrations were determined in the sheath cells and axoplasm of the nerve fiber of the squid Sepioteuthis sepioidea. The sheaths were obtained by slitting the nerve fiber, the extracellular electrolytes were washed out in isotonic sucrose solution, and the concentrations in the cells were determined after different soaking times in the sucrose solution. Values for the Schwann cell were calculated by extrapolation to zero time from the plots of the logarithms of the concentrations in the cells as a function of soaking time in sucrose solution. The Schwann cells made up 84 per cent of the sheath's total cellular volume. The Schwann cell concentrations in millimols per liter, are: 312 (404-241) for sodium, 220 (308-157) for potassium, and 167 (208-138) for chloride. The concentrations in the axoplasm (mean +/- SE), in millimols per liter are: 52 +/- 10 for sodium, 335 +/- 25 for potassium, and 135 +/- 14 for chloride. The possibility that some fraction of the Schwann cell electrolytes, especially of sodium, is bound, cannot be discarded.