Angiotensin II binding sites in individual segments of the rat nephron.

The sites of action of angiotensin II along the nephron are not well defined and both proximal and distal effects are suggested. Using a microassay that permits measurement of hormone binding in discrete tubule segments, we determined the binding sites of 125I-angiotensin II along the nephron of Sprague-Dawley rats. Specific binding in proximal convoluted tubule (PCT) (at 25 degrees C, pH 7.4) was linearly related to tubule length and saturable, with an apparent maximal binding capacity of approximately 300 amol X cm-1. Binding specificity was verified in competition experiments that revealed significant (P less than 0.001) and comparable competition for radioligand binding by angiotensin II and angiotensin precursor, metabolite, and analogues, whereas unrelated peptides of similar size (bradykinin, ACTH [1-10]) were without effect. The profile of specific angiotensin II binding along the nephron was: PCT, 216 +/- 13; pars recta, 86 +/- 14; medullary thick ascending limb of Henle's loop, 46 +/- 8; cortical thick ascending limb of Henle's loop, 77 +/- 8; distal convoluted tubule, 49 +/- 10; cortical collecting tubule, 15 +/- 1; medullary collecting tubule, 32 +/- 7 amol X cm-1. These results indicate the presence of specific angiotensin II binding sites in all tubule segments studied, but binding capacity was highest in the proximal convoluted tubule, in agreement with transport studies that localize the effects of the hormone in this segment.

Modulation of renal sodium-potassium-adenosine triphosphatase by aldosterone. Effect of high physiologic levels on enzyme activity in isolated rat and rabbit tubules.

The purpose of this study was to determine the nephron site, time course, and mechanism of mineralocorticoid action on renal tubular Na-K-ATPase in rats and rabbits, without dietary manipulation and by using the natural mineralocorticoid aldosterone. Sustained, high physiologic levels of circulating aldosterone mimicking those produced endogenously during potassium loading or sodium deprivation were provided by constant delivery of the hormone in doses of 5 or 50 micrograms/100 g body wt per 24 h, respectively, from osmotic minipumps implanted subcutaneously. In adrenal-intact rats receiving the 5-microgram dose, aldosterone levels were similar to those seen in animals fed a high K diet and produced a time-dependent increase in Na-K-ATPase activity in the cortical-collecting tubule (CCT) to a level 103% higher than in controls after 7 d (2,007 +/- 178 vs. 989 +/- 72 pmol/mm per h, P less than 0.001); the enzyme activity in the proximal convoluted tubule, medullary thick ascending limb, and the inner stripe of the medullary-collecting tubule did not change significantly. The increment in CCT Na-K-ATPase was larger (142%) in animals receiving for the same period of time the 50-micrograms dose, which produced circulating aldosterone levels similar to those of sodium-deprived rats. A significant stimulation of Na-K-ATPase activity was seen in the CCT of adrenalectomized rats after 24 h of treatment with either dose of the hormone, and at 12 h only in animals receiving the 50 micrograms/100 g per 24 h regimen. To determine whether the enhanced Na-K-ATPase activity produced by aldosterone is due to synthesis of new enzyme units or to alteration in its kinetics, we examined the ouabain-binding capacity and the affinity for Na and K of the enzyme from CCT of rabbits treated with 5 micrograms/100 g body wt per 24 h aldosterone for 3 d. These experiments revealed a parallel increment on Na-K-ATPase activity and specific [3H]ouabain binding in aldosterone-treated rabbits, while the affinity of the enzyme for either sodium or potassium was unaltered. The results of this study indicate that high physiologic levels of aldosterone simulating those measured during K loading or Na deprivation lead to a segment-specific increase in Na-K-ATPase activity in the CCT. This effect was time-and dose-dependent and was due to an increase in the number of active enzyme units. The segmental specificity and time course of the increase in enzyme activity suggest that modulation of Na-K-ATPase by aldosterone plays a role in the chronic adaptation of the CCT to altered availability of sodium and potassium, and therefore in the homeostasis of these cations by the kidney.

Regulation of renal Na-K-ATPase in the rat. Role of the natural mineralo- and glucocorticoid hormones.

Both mineralo- and glucocorticoids stimulate renal Na-K-ATPase, but their relative role in the regulation of the enzyme remains controversial. In this study we measured Na-K-ATPase activity in the cortical collecting tubule (CCT) of adrenalectomized rats replaced with either the native mineralocorticoid (aldosterone) or glucocorticoid (corticosterone) in doses calculated to yield previously determined physiologic concentrations of these hormones (5 ng X dl-1 and 5 micrograms X dl-1, respectively). This was achieved by continuous delivery of aldosterone (1 microgram X 100 g-1 X d-1) from an osmotic minipump or of corticosterone (2 pellets of 20 mg each), implanted subcutaneously either at adrenalectomy or 7 d later, when Na-K-ATPase activity reached its nadir. Adrenalectomized rats not receiving hormone replacement and adrenal-intact animals served as controls. The CCT was chosen because it contains the highest concentration of binding sites for both hormones. Na-K-ATPase activity declined 52% in the CCT of untreated adrenalectomized rats after 7 d, and remained unchanged thereafter. Physiologic replacement doses of aldosterone prevented this decline and restored the activity of the enzyme after it had been allowed to decrease maximally following adrenal ablation, whereas similar replacement of corticosterone was without effect. These observations suggest that under physiologic conditions Na-K-ATPase in the CCT, a probable target nephron segment of both hormones, is under mineralocorticoid rather than glucocorticoid control.

Gaps in the anion gap.

OBJECTIVE: To review limitations of the use of serum anion gap in clinical practice. DATA SOURCES: Original reports and reviews. STUDY SELECTION: Sources containing the most recent pertinent information. DATA SYNTHESIS: Theoretical and practical limitations beset the use of serum anion gap. Awareness of these limitations reduces but does not eliminate wrong diagnoses based on the anion gaps. CONCLUSIONS: Serum anion gap has a limited value in the differential diagnosis of acid-base disorders and can be misleading.

Hypertension in obese patients: hemodynamic and volume studies.

Distinct hemodynamic and volume characteristics have been suggested for established hypertension in severe obesity, namely, a high cardiac output, and expanded blood volume, and a normal peripheral resistance. To evaluate whether hypertension in moderately obese patients represents a separate entity that can be defined by hemodynamic and volume profiles, we studied these in 50 such patients and compared results with those obtained in 59 nonobese essential hypertensives and 25 normal subjects. Both obese and nonobese hypertensives had a normal cardiac index (men, 2.8 +/- 0.1 vs 2.8 +/- 0.09 liter/min/m2; women, 2.9 +/- 0.1 vs 2.8 +/- 0.1 liter/min/m2, respectively) and similarly elevated total peripheral resistance (men, 47.1 +/- 2.3 vs 46.5 +/- 1.9 U . m2; women, 45.0 +/- 2.4 vs 44.0 +/- 1.3 U . m2, respectively) as compared to normals (cardiac index: men, 2.9 +/- 0.09 liter/min/m2, women, 3.4 +/- 0.2 liter/min/m2; total peripheral resistance: men, 29.4 +/- 1.0 U . m2, women, 28.3 +/- 2.8 U. m2). Volume measurements corrected to body surface area showed that both obese and nonobese hypertensive patients had lower blood volume (men, 2.6 +/- 0.05 vs 2.5 +/- 0.05 liter/m2; women, 2.2 +/- 0.05 vs 2.3 +/- 0.05 liter/m2, respectively) than normals (men, 2.9 +/- 0.08 liter/m2; women, 2.5 +/- 0.08 liter/m2). The results of this study suggest that hypertension in moderately obese subjects is similar in its hemodynamic and volume profiles to hypertension in the nonobese and that the presence of obesity does not alter the hemodynamic characteristics of established essential hypertension.

Responses of the stenosed and contralateral kidneys to [Sar1, Thr8] AII in human renovascular hypertension.

To better define the intrarenal hemodynamic effects of angiotensin in human renovascular hypertension, 10 patients underwent renal hemodynamic and functional measurements before and during infusion of a competitive angiotensin analog, [Sar1, Thr8] AII. Eight had technically satisfactory split function studies. Despite a fall in mean arterial pressure (132 +/- 6 to 121 +/- 6 mm Hg, p less than 0.05) and humoral changes consistent with angiotensin-mediated hypertension, the intrarenal effects of this analog were commonly those of an angiotensin agonist, producing vasoconstriction and sodium retention. This was quantitatively greatest in the contralateral kidney, whose preinfusion sodium excretion (86 +/- 30 microEq/min vs 25 +/- 9 microEq/min, p less than 0.02) and glomerular filtration rate (76 +/- 7 ml/min vs 41 +/- 7 ml/min, p less than 0.01) were higher than the stenotic kidney. In some cases, an increase in renal blood flow and rise in sodium excretion were evident during angiotensin blockade, suggesting a tonic intrarenal action of angiotensin. Although renin vein renin values differed markedly between the stenotic and contralateral kidney (ratio = 2.05 +/- 0.30), relative changes in effective renal plasma flow were correlated (r = 0.84: p less than 0.01) during infusion of this analog. These results underscore the differences in sensitivities between vascular beds to the effects of angiotensin II and the major role of the contralateral kidney in renal function and sodium homeostasis in human renovascular hypertension.

Course of renal pathology in patients with systemic lupus erythematosus.

Evaluation of the course of lupus nephropathy by serial kidney biopsy in 50 patients revealed a complex pattern of transitions from one histologic class to another. A high rate of transformations (56 percent) was observed, with fewer than half the patients remaining in the original category. Although the general trend was towards transformation to a less severe lesion (WHO classes III and IV transforming into classes II and V), this was certainly not the rule for all individual classes. These transformations were rarely predictable on the basis of available clinical, laboratory, or pathologic information, and were less common in younger patients. These results help clarify the pathologic behavior of lupus nephropathy in the modern therapeutic era and highlight the value of pathologic examination for the planning and evaluation of therapy in selected patients.

Nondiabetic renal disease in patients with diabetes mellitus.

Renal diseases other than diabetic nephropathy were found in 10 of 122 diabetic patients who underwent renal biopsy between 1960 and 1982. These diseases included lupus glomerulonephritis, acute post-streptococcal glomerulonephritis, membranoproliferative glomerulonephritis (type I), focal glomerulosclerosis, idiopathic membranous nephropathy, and nonspecific immune complex glomerulonephritides. Because some of these disorders can alter the management and prognosis of renal disease in diabetic patients, the appearance of urinary abnormalities or deterioration in renal function inconsistent with the natural history of diabetic nephropathy raises the possibility of a nondiabetic renal disease and should lead to a more detailed evaluation.

Erythropoietin is produced by tubular cells of the rat kidney.

The cellular site of erythropoietin (epo) production within the mammalian kidney is still not completely understood. In the present study, we examined the expression of epo mRNA in microdissected rat nephron segments by RT-PCR after induction of epo expression with cobalt chloride. Erythropoietin mRNA was not detected in nephron segments from saline injected rats. In cobalt chloride injected animals, epo mRNA was found in the majority of samples from the cortical region of the nephron, PCT, and CAL. Medullary tubule preparations (MCT and MAL) were mostly negative for epo mRNA, and glomeruli were uniformly negative. The induction of epo transcripts in tubular cells by cobalt chloride was paralleled by stimulation of the major transport enzyme in the kidney, namely, Na-K ATPase in a tubular profile similar to that of induction of epo transcripts. These results support some earlier findings that epo gene expression in response to cobalt salt stimulation of rat kidney occurs in transporting tubular epithelial cells.

Transport enzymes and renal tubular acidosis.

By analogy to the findings with other transport disorders such as Bartter's or Liddle's syndrome, it might be expected that the various forms of renal tubular acidosis (RTA) could result from defects in H-ATPase or H-K-ATPase. However, the available data do not yet support such a simple explanation. With regard to distal RTA, inhibition of H-K-ATPase with inhibitors such as vanadate blocks the increase in enzyme activity observed with potassium depletion, but does not produce distal RTA. H-K-ATPase does not increase with metabolic acidosis, and inhibition of its activity does not decrease ammonium or total acid excretion unless K depletion is also present. Maleic acid administration produces proximal RTA along with other proximal tubular dysfunction in experimental animals. However, it acts by reducing Na,K-ATPase activity rather than by affecting specific H+ ion transporters. This is pertinent to the findings that Na,K-ATPase activity is reduced in obstructive uropathy. Although the acidification defect in this disorder has been ascribed to a defect in H-ATPase, Na-K-ATPase function is also impaired. Thus, the role of isolated defects in H+ transporters in the development of clinical acidification disorders remains to be elucidated.

Reversal of left ventricular hypertrophy with captopril: heterogeneity of response among hypertensive patients.

Reversal of left ventricular hypertrophy (LVH) has been reported not to occur with all antihypertensive agents. Moreover, a dissociation between blood pressure response to medical therapy and reversal of ventricular hypertrophy has been previously observed. To evaluate the effects of captopril we studied the electrocardiographic (ECG) changes in 26 severe hypertensive patients who received the drug for more than one year. In 14 patients with normal pretreatment ECG, captopril controlled blood pressure effectively [132 +/- 2.9 (SE) to 104 +/- 3.9 mmHg, p less than 0.001], but had no effect on ECG voltage. In 12 patients with pretreatment LVH, two different response patterns were observed despite similar blood pressure control (144 +/- 4.9 to 102 +/- 3.1 mmHg and 148 +/- 7.3 to 109 +/- 7.3 mmHg, p less than 0.001 for both): seven had complete normalization of ECG while five had residual LVH pattern. No significant difference was found between the latter two groups in regard to age, sex, weight, etiology of hypertension, pretreatment ECG voltage, blood pressure, plasma renin activity, duration of treatment and duration of maintained blood pressure control. The reversal of LVH pattern occurred early (between 12 to 16 months) with no overall correlation between lowering of blood pressure and ECG voltage changes. The heterogeneity of response observed in this study suggests that factors other than blood pressure control modify the reversal of cardiac hypertrophy by antihypertensive therapy.

Correlation between peritoneal equilibration test and dialysis adequacy and transport test, for peritoneal transport type characterization. Mexican Nephrology Collaborative Study Group.

OBJECTIVE: The aim of this study was to analyze the correlation between the peritoneal equilibration test (PET) and the dialysis adequacy and transport test (DATT) for peritoneal transport type characterization, and the degree of patients' acceptance for each test. DESIGN: Cross-sectional, observational multicenter study. SETTING: Five referral (tertiary) dialysis centers of institutional practice. PATIENTS: The study included 107 adult continuous ambulatory peritoneal dialysis (CAPD) patients with a prescription of four exchanges of 2 L per day, irrespective of age, gender, cause of end-stage renal disease, time on dialysis, nutritional status, or residual renal function. Patients on immunosuppressive therapy and those with cancer, hepatitis B, or HIV, and those having a peritonitis episode within the previous 30 days, or three or more episodes during the previous 12 months, were excluded. MAIN MEASURES: Peritoneal transport type as classified by creatinine and urea dialysis-to-plasma (D/P) ratios by PET and DATT. RESULTS: Correlation coefficients between D/P ratios for creatinine and urea, obtained for the PET and the DATT, were 0.73 for D/P creatinine and 0.96 for D/P urea. Patients were classified into high, high-average, low-average, and low transport categories according to the mean and standard deviation of D/P creatinine values obtained from the PET at 4 hours. These values showed excellent concordance with those generated from the DATT data (alpha = 0.82, 95% confidence interval 0.67 - 0.93). Nineteen percent of patients showed discordance in their category when classified according to the PET versus the DATT. Patients' acceptance was better for the DATT than for the PET, as evaluated with a questionnaire. CONCLUSION: The DATT is an easy, inexpensive, and reliable test to assess peritoneal transport type, and it also provides information about peritoneal clearance of solutes and ultrafiltration. The DATT has better patient acceptance than the PET. Since the DATT has only been validated for patients on a fixed CAPD daily schedule of 4 x 2 L, the results should be confined only to patients receiving such a prescription.

Proarrhythmic safety of repeat doses of mirabegron in healthy subjects: a randomized, double-blind, placebo-, and active-controlled thorough QT study.

Potential effects of the selective ß(3)-adrenoceptor agonist mirabegron on cardiac repolarization were studied in healthy subjects. The four-arm, parallel, two-way crossover study was double-blind and placebo- and active (moxifloxacin)-controlled. After 2 baseline ECG days, subjects were randomized to one of eight treatment sequences (22 females and 22 males per sequence) of placebo crossed over with once-daily (10 days) 50, 100, or 200 mg mirabegron or a single 400-mg moxifloxacin dose on day 10. In each period, continuous ECGs were recorded at two baselines and on the last drug administration day. The lower one-sided 95% confidence interval for moxifloxacin effect on QTcI was >5 ms, demonstrating assay sensitivity. According to ICH E14 criteria, mirabegron did not cause QTcI prolongation at the 50-mg therapeutic and 100-mg supratherapeutic doses in either sex. Mirabegron prolonged QTcI interval at the 200-mg supratherapeutic dose (upper one-sided 95% CI >10 ms) in females, but not in males.