Intestinal transport of weak electrolytes. Evidence in favor of a three-compartment system

A study has been made of the transmural fluxes of benzoic, phenylacetic, and pentanoic acids, benzylamine, hexylamine, and D-amphetamine across rat jejunum incubated in vitro. The M to S fluxes of the weak acids were greater than their corresponding S to M fluxes, and the S to M fluxes of the weak bases were larger than their M to S fluxes. These patterns of asymmetric movements were observed when the transmural electrical potential difference was clamped at 0 mV, and when the pH values of the mucosal and serosal fluids were identical. The effects of a weak acid on the fluxes of other weak electrolytes were qualitatively similar when the effector weak acid was added to the mucosal fluid, and when it was added to the serosal fluid. But the effects of a weak base on the fluxes of other weak electrolytes were dependent upon its location, and the interactions observed when the effector weak base was added to the mucosal fluid were qualitatively different than those seen when it was added to the serosal fluid. The interactions between weak electrolytes could readily be explained in terms of the function of a system of three compartments in series, in which the pH of the intermediate compartment is greater than that of the bulk phases. But these observations could not be explained in terms of an analogous system involving an intermediate compartment of low pH, or in terms of a carrier mediated system. The transport function of the three-compartment system can be described in the form of an equation, and it is found that a pH difference of less than 0.5 unit may explain our observations on weak electrolyte transport.     

Sodium influx at the outer surface of frog skin

Summary The unidirectional sodium influx across the outside surface of the frog skin epithelium was measured. The method was identical to the one described by Biber and Curran [4] except that mannitol instead of inulin was used as the indicator for the amount of tracer containing test solution remaining on the surface of the skin after blotting. The space of distribution for³H-mannitol is about twice as large as the corresponding space for inulin after 32-sec exposure to the tracer. At a sodium concentration of 6.7mm the sodium influx determined with inulin as marker for the test solution is 0.146 Equiv hr^–1cm^–2 larger than the influx measured in the same preparation with mannitol. This difference increases in proportion to the sodium concentration and can be ascribed to diffusion of sodium into a space which is accessible to mannitol but not to inulin during a 30-sec interval. Hence, nearly two-thirds of the previously described linear component of the sodium influx proceeds, as was suspected previously, into a compartment which is not directly connected to net sodium transport across the skin. The nature of the remaining one-third of this linear compartment is not clear but previous experiments suggest that it is also not involved in net sodium transport.     

Prediction of LDL concentration at the luminal surface of a vascular endothelium

To find out whether concentration polarization of low-density lipoprotein (LDL) occurs at the surface of a vascular endothelium or not, transport of LDL in flowing blood to an water-permeable endothelium was studied theoretically by means of CFD. Calculations were carried out for an endothelium exposed to a Couette flow by assuming that the surface geometry of the endothelium could be expressed by a cosine function. Two typical cases were considered for the permeability of endothelium to water; one was uniform permeability everywhere in the endothelium, and the other was uneven permeability which was augmented at the intercellular junction. It was found that, in both cases, the surface concentration of LDL increased in going distally from the entrance, taking locally high and low values at the valleys and hills of the endothelium, respectively, and the variation was larger in the case of endothelium with uneven permeability. These results clearly showed that concentration polarization of LDL which might affect the uptake of LDL by the arterial wall certainly occurs at the surface of the endothelium even if the flow is disturbed microscopically by the uneven surface of the endothelium.     

Estrogen action at endometrial membranes: alterations in luminal surface detectable within seconds

The morphological effects of estrogen on the luminal surfaces of rat endometrial cells were investigated by scanning electron microscopy. Ovariectomized rats were injected intravenously with estradiol-17 beta (E2 beta), 0.5 micrograms/0.25 ml per 100 g body wt. At various intervals thereafter, the lumen of a uterine horn was flushed with buffered 2% glutaraldehyde and then prepared for scanning electron microscopy by conventional methods. In control rats that had received an equivalent volume of placebo vehicle, the luminal cell surface was characterized by short, sparse microvilli (MV) and, in most cells, a single, central cilium. At 30 s after E2 beta injection, the number of MV was significantly increased. By 1 min, MV density was further increased and MV were frequently clustered; also, the central cilium of many cells was no longer evident. Similar results were obtained after exposure to diethylstilbestrol for 30 s to 1 min, whereas neither a subthreshold dose of E2 beta nor a dose of the relatively inactive congener E2 alpha equivalent to a saturating concentration of E2 beta gave statistically significant responses in surface changes by the present criteria. After 3-7 min of E2 beta exposure, MV had increased greatly in length and density. These effects underwent dramatic regression by 15-30 min after E2 beta treatment, with distinct diminution of microvillar lengths and numbers, reduction of clustering, and reappearance of the central cilium in many cells. This was succeeded at 1 h by a renewed surge of surface activity. These results are consistent with cumulative evidence for rapid alterations of the surface membrane of estrogen-sensitive cells in response to physiological levels of active hormone. Whether these responses in the luminal surfaces are primary, or are secondary reflections of receptor- mediated membrane alterations at the basolateral blood-front, remains to be determined.     

Active transport at the blood-CSF barrier contributes to manganese influx into the brain

Manganese is an essential trace element, and a contrast agent of potential interest for brain magnetic resonance imaging. Brain overexposure to manganese, however induces a neurodegenerative syndrome. Imaging data suggest that manganese appearance into the CSF precedes its accumulation into the cerebral parenchyma. We therefore investigated manganese uptake and transport at the blood-CSF barrier. Like lead, the non protein-bound divalent manganese accumulated into the rat choroid plexus. The metal accumulation was especially high in developing animals. Using a differentiated cellular model of the blood-CSF barrier, we demonstrated that manganese crosses the choroid plexus epithelium by a concentrating, unidirectional blood-to-CSF transport mechanism. This transport was inhibited by calcium, which is also transported into the CSF against its concentration gradient. The permeability barrier function towards lipid-insoluble compound and the organic anion transport property of the blood-brain interface were affected by exposure of the blood-facing membrane of choroidal cells to micromolar concentrations of manganese, but its antioxidant capacity was not. The unidirectional transport of manganese across the choroid plexus provides the anatomo-functional basis linking the systemic exposure to manganese with the spreading pattern of manganese accumulation observed in brain imaging, and explains the polarized sensitivity of choroidal epithelial cells to manganese toxicity.     

Flow-dependent concentration polarization of plasma proteins at the luminal surface of a semipermeable membrane

The effect of steady shear flow on concentration polarization of plasma proteins and lipoproteins at the luminal surface of a semipermeable vessel wall was studied experimentally using suspensions of these molecules in a cell culture medium and a semipermeable membrane dialysis tube which served as a model of an implanted vascular graft or an artery. The study was carried out by flowing a cell culture medium containing fetal calf serum or bovine plasma lipoproteins or bovine albumin through a 7.5 mm diameter, 60 mm-long dialysis tube in steady flow under a physiologic mean arterial perfusion pressure of 100 mmHg, and measuring the filtration velocity of water (cell culture medium) at the vessel wall which varied as a consequence of the change in concentration of plasma protein particles at the luminal surface of the semipermeable membrane dialysis tube. It was found that for perfusates containing plasma proteins and/or lipoproteins, filtration velocity of water was the lowest in the absence of flow, and it increased or decreased as the flow rate (hence wall shear rate) increased or decreased from a certain non-zero value, indicating that surface concentration of protein particles varied reversibly as a direct function of flow rate. It was also found that at particle concentrations equivalent to those found in a culture medium containing serum at 5% by volume, plasma lipoproteins which were much smaller in number and lower in concentration but larger in size than albumin, had a much larger effect on the filtration velocity of water than albumin. These findings were very much the same as those previously obtained with a cultured endothelial cell monolayer, strongly suggesting that the flow-dependent variation in filtration velocity of water at a vessel wall results from a physical phenomenon, that is, flow-dependent concentration polarization of low density lipoproteins at the luminal surface of the endothelial cell monolayer.     

Determinants of cation transport selectivity: Equilibrium binding and transport kinetics

ConclusionThe equilibrium ion-binding properties of ion channels and transporters can be difficult to discern from crystal structures alone, as proteins often adopt different lowest energy states depending on the ions bound. In cases where transport is slow, their inherent ion-binding preferences can be used to infer their transport preferences. However, in cases where transport is fast, the transport selectivity can hide their equilibrium preferences by accentuating the kinetics of ions hopping through a channel over its inherent ion-binding preferences. Thus, depending on the arrangement of ion-binding sites in a channel’s selectivity filter, one can achieve either selective or nonselective ion transport.The equilibrium K⁺ selectivity of some nonselective channels suggests a potential mechanism whereby they could evolve into a fast K⁺-selective channel. K⁺ channels and nonselective channels like CNG and HCN are related to one another in both sequence and structure, suggesting an evolutionary link between them. Swap experiments show that only a few mutations separate a nonselective channel from a K⁺-selective channel. One might imagine an evolutionary path between these channels in which the equilibrium preference for a K⁺ ion in a nonselective channel evolves into a K⁺-selective channel through these few mutations to create the selective ion queue. Alternatively, a slow single-ion channel with an equilibrium and transport preference for K⁺ ions could be transformed into a fast multi-ion channel through mutations that create a queue of K⁺-selective ion-binding sites, as is seen in most K⁺ channels studied to date.In the case of multi-ion selectivity filters, such as those found in K⁺ channels, the selectivity filter can be viewed as the active site that interacts with different queues of ions and water molecules. At least three properties emerge from multi-ion queues: (1) high conductance by reducing the affinity of multiple bound ions versus single ions; (2) high selectivity by allowing disfavored ions time to dissociate back into solution; and, consequently, (3) robust selectivity in an environment where ion concentrations can change. For transporters and carriers, the equilibrium preference and slow transport naturally create robust selectivity. In all these cases, equilibrium-based ion selectivity is achieved by slowing transport enough so that the disfavored ion is able to dissociate back into solution before transport takes place.     

Determinants of Intestinal Detoxication of Lipid Hydroperoxides

It has long been recognized that hydroperoxides are agents of cytotoxicity. However, in recent years, it is increasingly apparent that lipid hydroperoxide may play an important role in mediating cellular and molecular events in degenerative pathophysiological processes that lead to intestinal disorders, such as cancer. Yet, surprisingly, little is known of the intestinal disposition of peroxidized lipids and of the metabolic factors that determine mucosal peroxide elimination. The present paper summarizes the evidence for the pivotal role of reductant (GSH and NADPH) availability in intestinal peroxide detoxication. This information will provide important insights into the relationship between luminal lipid hydroperoxides and intestinal GSH redox homeostasis, and is pertinent to understanding how dietary oxidants like lipid peroxides, can impact intestinal integrity with implications for genesis of gut pathology.     

Facilitated transport of urea across the gall-bladder luminal membrane.

Counterflow experiments demonstrate the existence of urea counter-transport on the epithelium luminal surface. This phenomenon disappears when 10(-4) M phloretin is added to the perfusion fluid. Moreover counterflow experiments made using thiourea as elicitor, demonstrate that the phenomenon is specific for the urea.     

Flow-dependent concentration polarization of plasma proteins at the luminal surface of a cultured endothelial cell monolayer

Flow-dependent concentration or depletion of atherogenic low density lipoproteins which has been theoretically predicted to occur at a blood/endothelium boundary may play an important role in the genesis, progression, and regression of atherosclerosis in man and intimal hyperplasia in vascular grafts implanted in the arterial system in man and experimental animals. Hence to explore such a possibility, we have studied the effect of a steady shear flow on concentration polarization of plasma proteins and lipoproteins at the luminal surface of a cultured bovine aortic endothelial cell (BAEC) monolayer which served as a model of the vessel wall of an artery or an implanted vascular graft. The study was carried out by circulating a cell culture medium containing fetal calf serum or bovine plasma lipoproteins in steady flow through a parallel-plate flow cell in which a cultured BAEC monolayer was installed, over the physiologic ranges of wall shear rate and water filtration velocity at the BAEC monolayer. The water (cell culture medium) filtration velocity at the BAEC monolayer was determined to provide a measure of the change in concentration of plasma protein particles at the luminal surface of the BAEC monolayer. It was found that for perfusates containing plasma proteins and/or lipoproteins, water filtration velocity varied as a function of flow rate, being lowest in the absence of flow. Water filtration velocity increased or decreased as flow rate increased or decreased from an arbitrarily set non-zero value, indicating that surface concentration of protein particles varied as a direct function of flow rate, and the process was reversible. It was also found that at particle concentrations equivalent to those found in a culture medium containing serum at 20% by volume, plasma lipoproteins which were much smaller in number and lower in concentration but larger in size than albumin, showed almost the same effect as observed with serum which contained both lipoproteins and albumin, indicating that the substance responsible for this phenomenon is not albumin but lipoprotein whose diffusivity is much smaller than that of albumin. The results strongly support our hypothesis that flow-dependent concentration polarization of lipoproteins occurs at a blood endothelium boundary, and this in turn promote the localization of various vascular diseases which develop in our arterial system.     

Magnesium transport in Salmonella typhimurium: characterization of magnesium influx and cloning of a transport gene

The influx of Mg2+ in Salmonella typhimurium LT-2 was studied by both kinetic and genetic techniques. Wild-type cells grown in a high MgSO4 concentration (10 mM) exhibited a Km of 15 microM for Mg2+ influx, with a Vmax of 0.25 nmol of Mg2+ per min per 10(8) cells. The apparent Km decreased to 3 microM, and the Vmax increased 60% after growth in a low MgSO4 concentration (10 microM). Co2+ was a simple competitive inhibitor (Ki = 30 microM) of Mg2+ influx in cells grown in high Mg2+ concentrations but blocked only a portion of the Mg2+ influx in cells grown in low Mg2+ concentrations. Co2+ influx exhibited kinetics similar to those of Mg2+ influx (Km = 30 microM; Vmax = 0.5 nmol of Co2+ per min per 10(8) cells) but was not affected by growth conditions. Co2+ influx was competitively inhibited by both Mg2+ and Mn2+. Mutations affecting Mg2+ uptake were isolated by selection for spontaneous resistance to toxic levels of Co2+. One class of mutants designated corA mapped at 84 min near metE with the following gene order: corA, metE, zie-3161::Tn10, pepQ. A second class designated corB mapped at 98 min near pyrB. Mg2+ influx was decreased in a corA mutant strain (relative to that of the wild type) when grown in high Mg2+ concentrations but was restored when grown in low Mg2+ concentrations. Co2+ transport was completely abolished by the corA mutation under all growth conditions. Recombinant plasmids carrying the corA region from either Escherichia coli K-12 or S. typhimurium complemented the corA mutation in S. typhimurium, restoring uptake of both Co2+ and Mg2+ and conferring sensitivity to Co2+. The S. typhimurium corA gene was localized to a restriction fragment of approximately 1.5 kilobases.     

Strain driven transport for bone modeling at the periosteal surface

Bone modeling and remodeling has been the subject of extensive experimental studies. There have been several mathematical models proposed to explain the observed behavior, as well. A different approach is taken here in which the bone is treated from a macroscopic view point. In this investigation, a one-dimensional analytical model is used to shed light on the factors which play the greatest role in modeling or growth of cortical bone at the periosteal surface. It is presumed that bone growth is promoted when increased amounts of bone nutrients, such as nitric oxide synthase (NOS) or messenger molecules, such as prostaglandin E₂ (PGE₂), seep out to the periosteal surface of cortical bone and are absorbed by osteoblasts. The transport of the bone nutrients is assumed to be a strain controlled process. Equations for the flux of these nutrients are written for a one-dimensional model of a long bone. The obtained partial differential equation is linearized and solved analytically. Based upon the seepage of nutrients out of the bone, the effect of loading frequency, number of cycles and strain level is examined for several experiments that were found in the literature. It is seen that bone nutrient seepage is greatest on the tensile side of the bone; this location coincides with the greatest amount of bone modeling.     

The luminal surface of the rat trachea during ontogenesis]

To complete the hitherto existing results about the trachea epithelium, the scanning electron microscope was used to get representative statements about the apical surface of the epithelium: First of all the epithelium consists of undifferentiated round cells with cytopodia. The cells rarely carry single cilia at the same time. At the 18. day of pregnancy nearly all of the cells have a single cilium. Until the end of the intrauterine phase the single cilia are retracted again. At the 20. day ciliated cells and cells with protrusions are formed. Mucus granula are secreted into the lumen already before birth. Immediately post partum much mucus appears. Its distribution, viscosity and optical behaviour is different. For ciliated cells and goblet cells no special characteristical distribution was noticed. The building of the cells surface was detailed discussed. The question about function of the single cilia cannot yet be answered.