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L J Mandel

Publications and source records attributed to L J Mandel.

120 records · Page 7Linked to original sources
PubMed1978

Effects of pH, Ca, ADH, and theophylline on kinetics of Na entry in frog skin.

The short circuit current as a function of Na concentration in both solutions was found to obey Michaelis-Menten kinetics under a variety of experimental conditions. Values of maximal transport rate (Im) and half-maximal Na concentrations (Kt) were determined from these experiments. Three type of results were obtained: 1) Im and Kt both decreased by approximately the same fraction when the pH of both solutions was reduced by increasing PCO2, 2) Im decreased and Kt increased when the external pH was decreased, and 3) Im increased with ADH and theophylline, decreased with external Ca, and Kt remained unchanged. Various criteria were utilized to determine that these were properties of the entry barrier for Na into the "transport pool." The results are explained in terms of a model that separates three different types of actions on the entry barrier: 1) competition of Na with other ions in the external solution for entry, 2) modulation of the number of sites available for Na translocation by changing the cytoplasmic pH, and 3) alterations in the rate of Na translocation caused by changes in the Na permeability or the electrochemical gradient across the entry barrier.

Animals↗
PubMed1976

Effect of amiloride and some of its analogues of cation transport in isolated frog skin and thin lipid membranes.

The inhibition of short-circuit current (Isc) in isolated frog skin and the induction of surface potentials in lipid bilayer membranes produced by the diuretic drug amiloride and a number of its chemical analogues was studied. The major conclusions of our study are: (a) The charged form of amiloride is the biologically active species. (b) Both the magnitude of Isc and the amiloride inhibitory effect are sensitive to the ionic milieu bathing the isolated skin, and these two features are modulated at separate and distinct regions on the transport site. (c) Amiloride is very specific in its inhibitory interaction with the Na+ transport site since slight structural modifications can result in significant changes in drug effectiveness. We found that substitutions at pyrazine ring position 5 greatly diminish drug activity, while changes at position 6 are less drastic. Alterations in the guanidinium moiety only diminish activity if the result is a change in the spatial orientation of the amino group carrying the positive charge. (d) Amiloride can bind to and alter the charge on membrane surfaces, but this action cannot explain its highly specific effects in biological systems.

Amiloride↗
PubMed1975

Redox state of respiratory chain enzymes and potassium transport in silkworm mid-gut.

The midgut of Hyalophora cecropia actively transports potassium from hemolymph to lumen and the energy for this process appears to be intimately linked to oxidative metabolism. In the present investigation, we monitored concurrently the rate of active transport and the redox levels of the components of the respiratory chain in the intact tissue under a variety of experimental conditions. Approximately equal concentrations of cytochromes a3, a, c and b-557 were found. Other investigators (Pappenheimer, Jr, A.M. and Williams, C.M. (1954) J. Biol. Chem. 209, 915, Shappirio, D.G. and Williams, C.M. (1957) Proc. R. Soc. Lond. Ser. B 147, 233 and Chance, B. and Pappenheimer, Jr, A.M. (1957) J. Biol, Chem, 209, 931) have indentified cytochrome b-557 with b5 and found that it exists primarily in an extramitochondrial location. Steady-state experiments demonstrated that all these cytochromes were approximately 50% reduced while active transport proceeded at a high rate in regular cecropia Ringer containing 32 mM KCl. When the potassium concentration was reduced, the active transport decreased and all the cytochromes became more oxidized. Addition of 1 mM cyanide inhibited active transport by 90% and caused a 100% reduction of all cytochromes. Redox state and short circuit current (Isc) kinetics measured as the tissue was made anoxic showed that all the respiratory enzymes, except cytochrome b-557, became fully reduced at a faster rate than the rate of inhibition of the Isc. The rate of cytochrome b-557 reduction followed kinetically the Isc. These observations are interpreted in a scheme where cytochrome b-557 (possibly b5) branches off cytochrome c from the conventional resporatory chain, utilizing cytochrome a3 as the terminal oxidase for both branches. Cytochrome b-557 may be involved in providing a direct link between oxidative metabolism and active transport in the midgut of the silkworm.

Amobarbital↗
PubMed1975

Dilantin-calcium interaction and active Na transport in frog skin.

The effects of diphenylhydantoin (DPH) and calcium were simultaneously studied on solute transport in frog skin. In the presence of external Ca, DPH elicited a significant increase in active Na transport; in its absence, DPH had no significant effect on active transport. DPH increased passive permeability, but this action was independent of Ca. It appears that the increase in active transport elicited by DPH is due to a specific interaction with Ca at the transport site.

Animals↗
PubMed1975

Actions of external hypertonic urea, ADH, and theophylline on transcellular and extracellular solute permeabilities in frog skin.

Increases in transepithelial solute permeability were elicited in the frog skin with external hypertonic urea, theophylline, and vasopressin (ADH). In external hypertonic urea, which is known to increase the permeability of the extracellular (paracellular) pathway, the unidirectional transepithelial fluxes of Na (passive), K, Cl, and urea increased substantially while preserving a linear relationship to each other. The same linear relationship was also observed for the passive Na and urea fluxes in regular Ringer and under stimulation with ADH or 10 mM theophylline, indicating that their permeation pathway was extracellular. A linear relationship between Cl and urea fluxes could be demonstrated if the skins were separated according to their open circuit potentials; parallel lines were obtained with increasing intercepts on the Cl axis as the open circuit potential decreased. The slopes of the Cl vs. urea lines were not different from that obtained in external hypertonic urea, indicating that this relationship described the extracellular movement of Cl. The intercept on the ordinate was interpreted as the contribution from the transcellular Cl movement. In the presence of 0.5 mM theophylline or 10 mU/ml of ADH, mainly the transcellular movement of Cl increased, whereas 10 mM theophylline caused increases in both transcellular and extracellular Cl fluxes. These and other data were interpreted in terms of a possible intracellular control of the theophylline-induced increase in extracellular fluxes. The changes in passive solute permeability were shown to be independent of active transport. The responses of the active transport system, the transcellular and paracellular pathways to theophylline and ADH could be explained in terms of the different resulting concentrations of cyclic 3'-5'-AMP produced by each of these substances in the tissue.

Animals↗
PubMed1973

Response of the frog skin to steady-state voltage clamping. II. The active pathway.

Active Na transport across frog skin was separated from passive Na movement utilizing urea influx as a measure of passive (shunt) permeability. In this manner, the response of the overall active Na transport system to an applied potential was determined over a range from +200 mV to -100 mV. Active Na transport displays saturation as a function of applied potential, and both the level of saturation and the potential at which it is achieved are functions of the Na concentration in the external solution. The saturation with potential appears to involve a different step in the transport process than the saturation of Na flux as a function of external Na concentration. The observations can be qualitatively described by either a one-barrier or two-barrier model of the Na transport system.

Animals↗
PubMed1972

Response of the frog skin to steady-state voltage clamping. I. The shunt pathway.

Properties of the shunt pathway (a pathway in parallel to the Na transport system) in frog skin have been examined. The permeability of this shunt to urea increases markedly when the skin is depolarized to -100 mv (inside negative) but hyperpolarization to +100 mv produces no change in urea permeability compared to short-circuit conditions. The permeability increase at depolarizing potentials is dependent on the external solute concentration and is considerably reduced by the presence of external Ca. Neither urea permeability nor its response to changes in potential difference are affected by complete inhibition of Na transport by ouabain. In ouabain-poisoned skins, movements of Na, K, Cl, and mannitol through the shunt change in parallel with urea movements. Ion fluxes under these conditions and their response to potential can be described by the constant field equation. The selectivity of the shunt is in the order Cl > urea > K > Na > mannitol and this order does not appear to be affected by the absolute magnitude of the shunt permeability. Arguments are presented suggesting that the pathway is mainly between cells and that its permeability may be affected by cell swelling.

Animals↗
PubMedDate not supplied

Fluctuating shear stress effects on stress fiber architecture and energy metabolism of cultured renal cells.

The project investigates the relationship between the external shear force and the actin cytoskeleton along with the metabolic changes occurring inside the cells due to this force. Anchorage-dependent Madin Darby canine kidney (MDCK) cells were placed in spinner flasks with paddle-type stirrers agitated at 20 rpm, where they experienced shear stress fluctuations from 0.02 to 0.27 dyn/cm2 in magnitude. Following fixation, permeabilization, and staining with rhodamine-phalloidin, the relative amounts and distribution of F-actin stress fibers in the 1 micron basal layer of the cells were visualized by confocal microscopy. These structures disappeared after 12-15 h of exposure to shear stress. Previous results showed that the stress fibers disappear, leading to loss of epithelial attachment, after only 1 h of starvation-induced energy depletion. Therefore, in this study, the energy metabolism of the cells was established by measuring adenosine triphosphate (ATP) levels at different time intervals. No statistical difference in ATP content was found between the shear-stressed cells and the controls, showing that shear stresses cause cytoskeletal reorganization by a mechanism other than ATP depletion.

Actins↗