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To determine whether the Na+-K+-2Cl- symport or the parallel Na+/H+ and Cl-/HCO3- antiports provide the dominant pathway for NaCl uptake into the ciliary epithelium. Both pathways are known to support NaCl entry from the stroma into the pigmented ciliary epithelial (PE) cells, after which Na+ and Cl- diffuse across the gap junctions into the nonpigmented ciliary epithelial (NPE) cells and are released into the aqueous humor.Rabbit iris ciliary bodies were preincubated in HCO3-/CO2-containing or HCO3-/CO2-free solutions before quick freezing, cryosectioning, dehydration, and electron probe x-ray microanalysis.The NPE and the PE cells contained more K and Cl when incubated with bicarbonate. Inhibition of carbonic anhydrase with 0.5 mM acetazolamide had little effect in HCO3--free medium but prevented the increase in Cl in both cell types in HCO3-/CO2 solution. Inhibition of the Na+-K+-2Cl- symport with 10 to 500 microM bumetanide caused Cl loss from both cell types in HCO3--free solution, but bumetanide produced a paradoxical increase in Cl and Na in HCO3-/CO2 solution. Together, acetazolamide and bumetanide resulted in significant Cl loss in HCO3--free solution and prevented the gains of Cl and Na in HCO3-/CO2 solution.The present results indicate that the dominant entry pathway of NaCl from the stroma into the ciliary epithelial syncytium is through an acetazolamide-inhibitable Cl-/HCO3 and a parallel Na+/H+ antiport. The dominant release pathways into the aqueous humor appear to be a Na+-K+-2Cl-symport, which can be outwardly directed under physiological conditions, together with the Na+/K+-exchange pumps and Cl- channels.

The response of the villus and crypt cells of the mouse jejunum to secretagogues has been assessed through measurements of cellular composition with x-ray microanalysis. In nonstimulated tissues the Na concentration ([Na]c) of the crypt cells was significantly less, and the K ([K]c) and Cl ([Cl]c) concentrations were significantly greater, than that of the villus cells. There was also a decreasing gradient of [Na]c and increasing gradient of [K]c from the villus tip to crypt base due to a greater number of cells with a high [Na]c and low [K]c in the upper regions of the villi. Theophylline (10 mmol L−1) stimulated a sustained increase in bumetanide sensitive short circuit current (Isc) and significantly decreased the [Na]c of the villus cells. Similar, but smaller changes were seen in the crypt cells. Changes in villus cell [Na]c reflected a reduction in the number of cells with a high [Na]c. Inhibition of the apical Na/H exchanger (1 mmol L−1 amiloride) had little effect on basal Isc and the subsequent addition of theophylline increased Isc to a comparable extent as seen without amiloride. However, after amiloride treatment the only change in cellular composition was a reduction in the [Cl]c of both crypt and villus cells, suggesting that both regions are involved in the secretory response. These data suggest that the dominant response of the jejunum to secretagogues is an inhibition of Na absorption via Na/H exchange in the villi and the secretory response is distributed throughout the crypt/villus axis.

Toad urinary bladder epithelial cells were incubated in Na Ringer's with the serosal surface of the epithelium clamped at either +50 mV, 0 mV (short-circuited) or -50 mV with respect to the mucosal surface. Following incubation, portions of tissue were coated with an external albumin standard and rapidly frozen. Cryosections were freeze-dried and cell composition determined by x-ray microanalysis. Cell water and ion contents were unaffected when tissues were short-circuited rather than clamped close to their open-circuit potential difference (+50 mV). Incubation with vasopressin at +50 mV, and under short-circuit conditions, caused Na uptake without cell swelling or gain in Cl. Clamping at -50 mV resulted in uptake of water and ions, with considerable variation from cell to cell. These variations in cell composition were exacerbated by vasopressin. The greater the increase in water content, the greater the rise in cell Cl. However, there was no consistent pattern to the associated changes in cation contents. Most cells gained some Na. In some cells, this gain was accompanied by an increase in K. In others, the gain of Na was predominant and cell K content actually fell. At -50 mV with ouabain, many of the cells also gained water. As was found in our earlier study with ouabain under short circuit conditions (Bowler et al., 1991), there was considerable variation in the extent of the Na gain and K loss; some cells were largely depleted of K while in others the K content remained relatively normal. These results indicate differences between granular cells in the availabilities in the plasma membranes of ion pathways, either as a consequence of differences in the numbers of such pathways or in their control.

Relationships between short-circuit current (I sc), cell Cl and the mechanism(s) of Cl accumulation in toad bladder epithelial cells were investigated. In serosal Cl-free gluconate Ringer, 80% of the cell Cl (measured by x-ray microanalysis) was lost over 30–60 min with an associated decrease in cell water content. Concomitantly, I sc fell to 20% of its initial value within 10 min but then recovered to 45% of its initial value despite continued Cl loss. With the reintroduction of Cl, cell Cl and I sc both recovered within 10 min. Serosal SITS (4 acetamido-4′-isothiocyano-stilbene-2,2′-disulfonate; 0.5 mm) plus bumetanide (0.1 mm), did not prevent the fall in I sc or the loss of cell Cl in gluconate medium, although they did inhibit subsequent recovery of I sc in this medium. They also prevented the recovery of I sc in Cl medium but not the reaccumulation of Cl by the cells. Although SITS and bumetanide did not prevent the loss or recovery of Cl, they modified the pattern of the ion changes. In their absence, changes in cellular Cl were twice that of the changes in measured cellular cations implicating basolateral Cl/HCO3 exchange in Cl movement. With SITS plus bumetanide present, changes of similar magnitude in Cl were associated with equivalent changes in cation, consistent with the inhibition of Cl/ HCO3 exchange.

The carbon dioxide produced by toad urinary bladders bathed on their mucosal surfaces by sodium Ringer solution and on their serosal surfaces by modified Leibovitz tissue culture medium was analysed by multiple regression on both sodium transport and time. The fractions contributed by metabolism related to transport and by basal metabolism were assessed, and the extent to which these might vary with time was determined. This analytical method, which improves the accuracy with which suprabasal metabolism is estimated, was used to examine the effects on metabolism of vasopressin, aldosterone, and mucosa-positive voltage-clamping. Vasopressin (0.05 u./ml), which on average increased sodium transport 2.9 times and concurrently increased the rate of carbon dioxide production in these transporting tissues, also altered the carbon dioxide production of non-transporting, amiloride-treated control hemibladders. For each hemibladder the ratio of sodium transported to suprabasal carbon dioxide produced after vasopressin was compared with that observed before vasopressin. Differences between the ratios were much reduced when the carbon dioxide productions of the paired transporting hemibladders were corrected for the effects of vasopressin on basal carbon dioxide production. With such analysis, it was confirmed that vasopressin did not alter the stoichiometry of sodium transport. A 30 mV, mucosa-positive voltage clamp, applied near the peak of the response to vasopressin, further increased both sodium transport and carbon dioxide production. No alterations of the ratio of sodium to suprabasal carbon dioxide were seen under these conditions where the maximal rate of active sodium transport in this tissue must have been approached. Active sodium transport was more than doubled some 4 h after adding aldosterone (10(-7) M). However, the related increase in suprabasal carbon dioxide production was greater than threefold. Therefore, whereas the stimulation resulting from vasopressin and voltage clamping had no effect on the ratio of sodium transported to suprabasal carbon dioxide produced, this ratio was reduced significantly by aldosterone. When the sodium transport of aldosterone-treated bladders was increased further by voltage clamping, the ratio of sodium transported to suprabasal carbon dioxide production remained at the reduced value. Sodium transport was increased by approximately 35% more when aldosterone-treated hemibladders were voltage clamped after vasopressin, the control paired hemibladders being exposed to vasopressin and voltage clamping alone.(ABSTRACT TRUNCATED AT 400 WORDS)

1. The production of CO2 in relation to sodium transport by toad urinary bladder has been examined in the absence of sodium from the serosal medium. 2. Replacement of serosal sodium by choline increased CO2 production without stimulating short-circuit current. Replacement of serosal medium by Tris did not have this effect. 3. With serosal sodium Ringer replaced by Tris Ringer, the ratio of sodium transported to CO2 produced was not altered significantly. 4. The results therefore suggest that there is no important recycling of sodium between the serosal medium and the transporting epithelial cells.

1. Thin slices (0.2-0.3 mm) of rabbit renal cortex have been incubated in isosmotic oxygenated acetate media at 25 degrees C with or without ouabain (10(-3) M), amiloride (2 x 10(-3) M) or iodoacetamide (10(-3) M). 2. Slices in normal isosmotic 146 mM-sodium-132 mM-acetate media swelled as reported previously (Cooke & Macknight, 1984). This swelling was not prevented by amiloride or by metabolic inhibition. 3. Slices in isosmotic 132 mM-choline-132 mM-acetate media gained much less water and were little affected by ouabain, amiloride or metabolic inhibition. Choline was able to substitute neither for sodium nor for potassium in activating preparations of renal cortical Na+-K+-ATPase in chloride or in acetate media. 4. Slices in isosmotic 20 mM-sodium-132 mM-acetate medium swelled nearly as much as did slices in normal sodium acetate medium. However, this swelling was impaired markedly by amiloride, by ouabain and by metabolic inhibition. 5. There was a direct correlation between medium sodium concentration and slice water content as sodium was increased from 1.25 to 30 mM in 132 mM-acetate media. However, up to a sodium concentration of 10 mM, amiloride (2 x 10(-3) M) completely prevented this increase in water content. 6. Increasing medium amiloride concentration from 10(-5) to 10(-3) M progressively inhibited cellular swelling in 10 mM-sodium-132 mM-acetate medium. It is concluded that, under these experimental conditions, the dominant pathway for hydrogen ion extrusion from the cells was via amiloride-sensitive sodium-hydrogen exchange. 7. The results are discussed in terms of a model which explains cellular swelling in acetate media in terms of (a) non-ionic diffusion of acetic acid across plasma membranes impermeable to the acetate anion, (b) removal from the cells of the hydrogen ion gained with the acetate by amiloride-sensitive sodium-hydrogen counter-transport and (c) subsequent extrusion of sodium from the cell accompanied by potassium uptake via the ouabain-sensitive Na+-K+-ATPase. 8. The results provide evidence for ion movements across the luminal plasma membrane of proximal tubular cells in rabbit renal cortical slices.

An experimental and analytical investigation has been conducted to establish correlation between thrust chamber design parameters and resultant combustion phenomena. The tests were made with self-impinging doublet injectors in a 15-in. -diameter pulse motor using Nitrogen tetroxide and monomethyl hydrazine. Pulse guns were used to disturb the combustion process and rate combustion stability. The analytical, vaporization-controlled, combustion models were used to compute the stability characteristics of the experimental thrust chamber at each operating condition. The stability model was then evaluated by comparing the hot-firing test results with the predicted stability characteristics. (Author)

A consecutive series of 435 patients with breast cancer followed from six to thirty years is presented. Radical mastectomy was the surgical procedure of choice and our experience indicated that the degree of axillary nodal involvement is a most important guide in establishing prognosis in this disease. The majority of recurrences became evident within the first five postoperative years, the next most hazardous period for the patient being the half decade between the fifth and tenth years after initial therapy. A second independent malignancy developed in 37 of the 275 pateints who died, and it was the cause of death in 7.6 per cent of this group. Among the 160 patients still living to date a second independent malignancy has already developed in 10. Bilateral breast cancer was observed thirty-eight times in this entire series of 435 patients, an incidence of 8.7 per cent.

Three activities in which students participate to demonstrate an unsaturated solution, a saturated solution and dynamic equilibrium, and a supersaturated solution.