Prokallikrein and kallikrein inhibitors in human urine.

Kallikrein activity (KA) and kallikrein inhibitors (KI) were evaluated in urine from normal subjects and essential hypertensives. Kallikrein inhibitors were almost completely removed by dialysis against water, confirming previous reports of their low molecular size. The negative relationship found between KA and KI in urine samples from 12 normal subjects suggests that KI might play a role in the modulation of KA excreted by the kidney. Heating (85 degrees C for 20 min) partially reduced (-52%) KI, indicating thermostable substances other than peptides as KI. When cations were evaluated as possible KI, by adding different salts to dialysed salt-free urine, only sodium was able to inhibit KA by 20% at a concentration not far from the physiological range. Trypsin added to dialysed urine produced a striking increase in KA without significant changes in Km and pH optimum. These findings, together with the observation that acid dialysis did not increase KA, strongly support the hypothesis that trypsin-activatable kallikrein might be a pro-enzyme. A lower urinary excretion of active kallikrein was found in 48 essential hypertensives when compared with 31 normotensive controls. However, trypsin-activatable kallikrein excretion was similar in the two groups, suggesting that low KA in hypertensives might not reflect a defective conversion of the pro-enzyme into the active form.

Active and inactive renin during acute reduction of renal perfusion pressure in essential hypertensives.

Arterial and renal venous active and inactive renin were studied in 5 patients with long established moderate hypertension following unilateral acute reductions of renal perfusion pressure (15% and 70% of control) by inflating a balloon catheter introduced into the right renal artery. This procedure failed to induce the expected release of active renin; total and inactive renin levels were also unchanged. On the contrary in a single normotensive patient smaller reductions of the renal perfusion pressure (-15% and -30%) were able to acutely increase the release of active renin with a concurrent conversion of inactive renin but without inducing blood pressure changes. These findings show that the renin pattern typical of unilateral renovascular hypertension, including the intrarenal activation of inactive renin, could be reproduced acutely in a normotensive subject. Moreover, a complete reversal of the above mentioned active and inactive renin pattern was observed in a recent onset renovascular hypertensive patient within 30 min from successful percutaneous transluminal dilation. The negative results observed in our hypertensive patients suggest that structural changes induced by the long duration of hypertension might have reduced the sensitivity of the baroceptors involved in renin release.

Natriuretic effect of acute nifedipine administration is not mediated by the renal kallikrein-kinin system.

Despite their vasodilating action, calcium antagonists increase renal sodium excretion. To ascertain whether renal kallikrein plays a role in the renal effects of calcium antagonists, nifedipine (N) (10 mg orally) or placebo (P) was given to 17 male patients with mild to moderate essential hypertension during a 6-h infusion of either saline (S) or aprotinin (A) (2 X 10(6) KIU in 200 ml of saline). Blood pressure (BP) and heart rate (HR) were measured every 10 min, and blood samples were taken at -10, 0, 30, 60, 120, 240, 360 min for plasma renin activity (PRA), creatinine, and osmolarity determinations. Urinary kallikrein, aldosterone, creatinine, and electrolytes were measured in 6-h urine collections. The acute administration of N induced a significant systolic BP (SBP) and diastolic (DBP) fall and a transient PRA increase that peaked at 30 min and were not modified by A infusion. Urinary volume (+47%), Na+ (+54%) and Cl- (+58%) excretion were significantly enhanced by N. There were less pronounced and statistically not significant increases in urinary excretion of Ca2+ (+38%) and K+ (+29%). Infusion of A did not interfere with the natriuretic effect of N. Our data do not support the hypothesis that the kallikrein-kinin system plays an important role in mediating the renal effects of nifedipine in humans.

Comparison of sublingual and oral captopril in hypertension.

The use of sublingual captopril has been recently suggested in hypertensive crisis on the assumption of a faster absorption and thus a more rapid effect on blood pressure than with the oral route. To verify this hypothesis we have compared the hypotensive effect of oral and sublingual captopril in 40 essential hypertensives who were randomly allocated to either route of administration. Captopril was administered orally dissolved in water or allowed to dissolve under the tongue. After 5, 10, 20, 30, 40, 60 and 90 minutes blood pressure, Plasma Renin Activity (PRA) and Angiotensin Converting Enzyme (ACE) were measured. No significant differences were found between the two groups in the time course of blood pressure decrease, PRA increase and ACE inhibition. The changes of the parameters studied was superimposable irrespective of the route of administration thus not supporting the hypothesis that sublingual captopril might be absorbed more rapidly.

Plasma clearance rate of 51Cr-EDTA provides a precise and convenient technique for measurement of glomerular filtration rate in diabetic humans.

It has not yet been fully clarified whether the plasma or renal clearance approach is the most reliable to investigate GFR in humans. The study presented here aimed to compare plasma decay with renal clearance of 51Cr-EDTA in 27 diabetic patients with patterns of renal function broadly dispersed in a wide range of values. Moreover, the comparison was also performed with renal clearance of nonlabeled iothalamate in a subgroup of 17 patients. A biexponential function was found to fulfill statistical and heuristic criteria for the modeling analysis of plasma 51Cr-EDTA decay with 19 samples after bolus intravenous 51Cr-EDTA injection. Individual GFR values from 51Cr-EDTA plasma clearance highly correlated with those from renal clearance (r2 = 0.977, P < 0.0001), but resulted on average about 2.5 mL.min-1.1.73 m-2 higher (66.8 +/- 6.5 mL.min-1.1.73 m-2 (mean +/- SE) versus 64.3 +/- 6.4, P < 0.02). This difference remained relatively constant from patients with normal renal function to those with impaired renal function, suggesting that the plasma clearance is slightly less accurate than renal clearance approach because of a constant extrarenal clearance rate. In the subgroup studied, a similar difference was found between GFR values from 51Cr-EDTA plasma clearance (84.7 +/- 7.3) and renal clearance of iothalamate (82.8 +/- 7.3), although not statistically significant (P = 0.4). Individual GFR values well correlated (r2 = 0.913, P < 0.0001). The precision and reproducibility of the experimental approaches were assessed by comparing three coefficients of variation: (1) CVb of the bolus injection, because of measurement errors; (2) CVc of the continuous infusion, which additionally includes errors of urine volume measurement and physiological variability in the same day; and (3) CVr of repeated measurements by using bolus injection, which also accounts for physiological variability in different days. CVc of iothalamate and 51Cr-EDTA infusions were 7.5 +/- 1.9% and 7.4 +/- 1.2% respectively. CVb and CVr of bolus injection of 51Cr-EDTA were 2.6 +/- 0.3% and 3.5 +/- 0.8% respectively. CVb and CVr of bolus injection of 51Cr-EDTA, but not CVc of iothalamate and 51Cr-EDTA infusions were twofold to tenfold lower than the percent yearly change reported in IDDM and NIDDM patients. More particularly, CVr was significantly less than CVc. In order to make the test less cumbersome, a reduced sampling schedule with seven samples was designed and validated. GFR measured with seven samples was 66.1 +/- 6.4 (P = 0.1 when compared with the full 19-sample schedule) with a CVb of 3.5 +/- 0.5%. This seven-sample protocol was not different from that obtained with the previously described simplified method of Brøchner-Mortensen (63.9 +/- 6.8, P = 0.16), yet yielding a statistically more accurate estimate (coefficient of variation for Brøchner-Mortensen method = 12.1 +/- 2.9, P = 0.004). Moreover, only bolus injection, along with modeling analysis of plasma clearance rate, allows the accurate measurement of the extracellular fluid volume, an important parameter in diabetic patients. It was concluded that the reduced seven-plasma sample protocol is able to detect as small as 4 to 5% changes per year in a single patient. Moreover, it provides precise and accurate estimate of GFR in diabetic patients with hyperfiltration, who are postulated to be at higher risk to develop renal damage.