A new model of genetic hypertension in rats with superficial glomeruli.

Despite renal involvement in the genesis of hypertension, the precise renal hemodynamic events prior to and during development of hypertension have not been obtainable by direct study in the available rat models of genetic hypertension. We have developed a model of genetic salt-sensitive hypertension in rats with superficial glomeruli, using the protocol described by Dahl to develop the Brookhaven model. Adult Munich Wistar rats were purchased from Charles River and bred in our laboratory. Offspring were allowed to mature without intervention. Systolic blood pressure and body weight were measured weekly to determine the normal longitudinal changes with increasing age for this strain. Balance studies were also carried out longitudinally, and during these studies plasma renin activity (PRA) was determined. When the rats were 16 weeks old, the availability of superficial glomeruli was assessed under microscopic examination and only those rats with numerous superficial glomeruli/field were mated. The offspring of these breeders were used as the zero generation (F0) of salt-sensitive rats, and were treated as described by Dahl. Systolic blood pressure, body weights and balance studies were carried out in this and in all subsequent generations. Sibling breeding was maintained and rats were bred for salt-sensitivity (systolic blood pressure greater than 155 mmHg at the age of 2 months) and availability of superficial glomeruli. Those rats with systolic blood pressure of less than 120 mmHg were bred in an attempt to develop a salt-resistant strain, and untreated Munich Wistar rats were bred to provide another control.(ABSTRACT TRUNCATED AT 250 WORDS)

Excitatory amino acids and epilepsy-induced changes in extracellular space size.

Convulsant and stimulus induced seizures are associated with Ca, Na, K and Cl concentration changes in the extracellular space (ES), which are a resultant of transmembrane ionic fluxes and of changes in the ES size. The ES decreases on average by 30% during a single seizure. An analysis of the causes of ES size changes reveal a large contribution from the spatial glia K buffer mechanism which may account for up to 60% of the ES decreases. NaCl and KCl uptake into cells as well as increases in intracellular osmolarity due to anaerobic glycolysis contribute less to the local cytotoxic edema but account for a net gain of osmotic active particle at the site of the focus. Excitatory amino acids such as glutamate, aspartate, N-methyl-D-aspartate (NMDA), kainate and quisqualate also lead to Na, Cl and eventually Ca uptake into cells and to release of K with dose dependent decreases in [Na]o, [Ca]o and [Cl]o, increases in [K]o and transient decreases in ES size by up to 80% which are possibly associated with a net reduction of osmotically active particles. The predominant cause for this cytotoxic edema is NaCl uptake into cells but spatial K buffering through glial cells also contributes to this type of edema. The possible consequences of the various ion movements and the changes in osmolarity as well as ES size for tissue vulnerability are discussed.

Supernormality: recent developments.

The major advance in our understanding of supernormality is the following hypothesis. Focal accumulations of extruded potassium ion in unstirred-clefts (invaginated extensions of the extracellular space in Purkinje fibers) cause threshold to fall by virtue of the early effects of hyperkalemia: lowering of threshold and acceleration of conduction. These effects rapidly dissipate in diastole as ionic equilibration takes place; threshold rises and assumes its late diastolic value. It is not known if this mechanism is applicable to the supernormality of specialized atrial fibers. Comparing the strength interval curves of supernormality and triggered automaticity the fall in threshold during the upstroke of the late after-depolarisation occurs later than the supernormal phase and is followed by a threshold rise (during the downstroke of the after-potential). Supernormality does not correspondingly show this biphasic change in the strength interval curve.

Pathophysiology of ischemic cell death: III. Role of extracellular factors.

The purpose of this study was to determine the effect on cell survival of extracellular changes that occur during ischemia, over and above the depletion of O2 and substrate. Rabbit retinas were deprived in vitro of both O2 and substrate, and then returned to control medium for 4 h before recovery was assessed by measuring protein synthesis, glucose utilization, and tissue water. Experimental conditions were altered in various ways during the period of O2 and substrate deprivation in order to modify the changes taking place in the interstitial fluid as a result of the failure of energy metabolism. When O2-free, substrate-free extracellular electrolyte solution was added to the retinas to reduce the ischemia-induced changes in the interstitial fluid, there was marked reduction in irreversible damage. But when energy-deprived retinas were exposed to retinas that had already been ischemic, or to interstitial fluid from ischemic retinas, there was an increase in irreversible damage. Removing Ca++ from the extracellular fluid during the period of energy deprivation increased the damage due to short deprivations in a restricted volume of extracellular fluid, but reduced the damage from longer deprivations in a large volume of extracellular fluid. The results demonstrate that several changes occur in the extracellular fluid during ischemia that significantly affect recovery.

Interstitial fluid dynamics in conscious renal hypertensive rats.

Interstitial fluid pressure (IFP) was measured in conscious rats with one-kidney one-clip Goldblatt hypertension using implanted capsules. No significant differences in IFP were found between rats with hypertension of 3, 28, or 56 days duration and the one-kidney controls. To estimate interstitial compliance in the 28-day hypertensive and control rats, hyperosmotic albumin solution was infused intravenously. The subsequent transfer of tissue fluid into the intravascular space was associated with increased diuresis and decreased IFP. The changes in tissue fluid volume (TFV) were estimated from the changes in plasma volume and urine output. In the hypertensive rats 3 h postinfusion, TFV decreased by 36.3 +/- 2.9 ml/kg, and IFP decreased from -1.2 +/- 0.2 to -4.7 +/- 0.6 mmHg. In the normotensive rats TFV decreased by 36.7 +/- 2.7 ml/kg, and IFP decreased from -1.2 +/- 0.2 to -4.3 +/- 0.4 mmHg. Interstitial compliance, estimated from the regression of delta IFP and delta TFV, averaged 7.3 +/- 1.2 and 8.1 +/- 0.7 ml.kg-1.mmHg-1 (P greater than 0.5) in the hypertensive and normotensive rats, respectively. These results suggested that the rats with one-kidney one-clip Goldblatt hypertension had normal interstitial fluid dynamics.

The effects of verapamil, quiescence, and cardioplegia on calcium exchange and mechanical function in ischemic rabbit myocardium.

The effects of verapamil (1 mg/liter, 2 x 10(-6) mol/liter), quiescence, and cardioplegia (K+ 16 mmol/liter, Mg2+ 16 mmol/liter) on calcium exchange and mechanical function during ischemia and reperfusion have been investigated in the rabbit interventricular septum at 32 degrees C. Calcium influx and efflux were recorded continuously with 47Ca2+ and 45Ca2+. After 60 minutes of total ischemia and reperfusion for 30 minutes, there was a net calcium gain of 4.9 mmol/kg dry tissue. Verapamil given before total ischemia reduced net calcium gain to 1.5 mmol/kg dry tissue (n = 5, P less than 0.03). When given only on reperfusion after total ischemia, or 10 minutes before reperfusion during low flow ischemia, verapamil did not affect calcium exchange. Cardioplegia begun 10 minutes before total ischemia reduced net calcium gain to 1.0 +/- 0.26 mmol/kg dry tissue (n = 6, P less than 0.001). Cardioplegia during the first 10 minutes of reperfusion, or lack of electrical stimulation during reperfusion, did not reduce calcium gain. Net calcium gain correlated with the maximum rise in resting tension and with the recovery of developed tension. In control experiments neither verapamil nor cardioplegia altered influx or efflux of slowly exchanging calcium. The cardioprotective effects of cardioplegia and the calcium channel blocker verapamil appear to be due to a reduction of myocardial work rather than to any specific direct action on calcium fluxes across the myocardial cell membrane.

An electron microscopic study of satellite-cells and regeneration in dystrophic mouse muscle.

Triceps and gastrocnemius muscles from dystrophic (129 Rej dy/dy) and normal mice were examined by electron microscopy at different stages in development for evidence of regeneration. Mitotic satellite cells were present only in dystrophic muscle. Myoblasts containing myofilaments, and multinucleate myotubes were observed within foci of regeneration. Approximately 50% of the myotubes showed features indicative of degeneration or abnormal development. These features included the presence of membrane whorls, and myofibrillar and sarcolemma breakdown. Quantitative studies suggest that the number of satellite cells is increased in dystrophic muscle. It is concluded that there are sufficient satellite cells in dystrophic mouse muscle to allow regeneration, and they are able to proliferate and form well differentiated myotubes. However, subsequent development of the myotubes can be ineffective, or 'abortive', reducing the regeneration capacity to the muscle.

Acute saline expansion lowers blood pressure of early DOCA salt hypertensive rats.

Changes in blood pressure and body fluids volume caused by an acute saline load were studied in 6 normal and 7 DOCA-salt early hypertensive rats having normal inulin space and plasma volume. After loading, blood pressure increased in the normal group (+23 mm Hg) and fell (-17 mm Hg) in the DOCA-salt group. Expansion of extracellular fluid volume was similar in both groups but expansion of plasma volume was twice as high in the DOCA-salt than in the normal group. The results are compatible with an inhibition of a volume-sensitive vasopressor mechanism possibly involving vasopressin.

An explanation of impaired solute mixing in extracellular fluid after hemorrhagic hypotension.

A theory is presented suggesting that the slowed rate of solute dispersion in the extracellular fluid (ECF) space during or after hemorrhagic shock is largely due to a depressed cardiac output, and that mixing may be improved by any fluid therapy that decreases the mean recirculation time for the solute. This theory was tested by comparing solute mixing rates after treatment of shocked anesthetized dogs with either protein-free Ringer solution with Ringer solution containing 5% albumin. The rate of mixing was determined by computer analysis of the disappearance of an ECF tracer from plasma. Compared with Ringer-treated dogs, the albumin-treated group had a faster rate of tracer equilibration, larger cardiac plasma output, and smaller, approximately normal ECF volume. It was concluded that the abnormal dispersion of solute within the ECF space seen during or after hypotension can be corrected simply by restoring the cardiac output, and that albumin-containing fluid may be more effective in this regard than crystalloid solution alone.

Current views on the mechanisms of pulmonary oedema.

Understanding of the causes of pulmonary oedema must be based on knowledge of the mechanism responsible for fluid exchange between the several compartments of the normal lung. Recent physiological studies have clarified the main features of these mechanisms. However in three areas knowledge is still incomplete--the magnitude of the hydrostatic and oncotic forces responsible for fluid movement within the lung, the means by which protein leaks across the wall of small pulmonary vessels and the routes by which fluid and protein pass between the interstitial tissues of the lung and the alveolar space. Further work is needed in these areas. On the basis of this physiological knowledge the mode of development of hydrostatic oedema, the role of lymphatics in pulmonary oedema, and the several stages of pulmonary oedema development that may culminate in alveolar flooding are now clearly understood. Knowledge is less complete about oedema due to increased vascular permeability. In some experimental models, such as alloxan, leakage is due to irreversible injury to the alveolar wall; in other models, including ANTU, oedema formation has been shown to depend upon minor and reversible changes in pulmonary vascular endothelium similar to those that cause exudate formation in areas of acute inflammation. In no instance is detailed information available of both the rate and magnitude of protein leakage and of the morphological basis of increased vascular permeability. Further work is required in this area. Present knowledge allows an adequate explanation of the changes that occur in many clinically important types of pulmonary oedema, including cardiac failure and neurogenic pulmonary oedema. Other types of oedema, notably that which may complicate traumatic shock or extrapulmonary sepsis and high altitude pulmonary oedema, are more complex and the details of their pathogenesis are still obscure.

Total effective compliance, cardiac output and fluid volumes in essential hypertension.

Total effective compliance, hemodynamic parameters, extracellular fluid volume, cardiopulmonary (CPBV) and total blood (TBV) volumes were determined in 32 men, including 14 normotensive controls and 18 sustained essential hypertensive patients. The effective compliance was calculated from the changes in central venous pressure recorded simultaneously with the changes in blood volume obtained after a rapid Dextran infusion. In normotensive controls, compliance was 2.08 +/- 0.09 ml/mm Hg/kg and was positively correlated with plasma (r = 0.79) and extracellular fluid (r = 0.84) volumes. In hypertensives, compliance was significantly reduced (1.49 +/- 0.06 ml/mm Hg/kg; P is less than 0.001) and was correlated negatively with the CPBV/TBV ratio (r = -0.75) and positively with the plasma volume/interstitial fluid volume ratio (r = 0.84). These results suggest that in normotensives, there is a regulatory mechanism between volume and compliance and that this contributes to maintaining filling pressure and cardiac output within normal ranges. In hypertensives, the reduced compliance could participate in the maintenance of normal values of cardiac output and extracellular fluid volume by influencing the partition of intravascular and extracellular fluid volumes.

changes in serum and urinary calcium during treatment with hydrochlorothiazide: studies on mechanisms.

Studies were undertaken in man to evaluate the roles of volume depletion and of the parathyroid glands in mediating the changes in serum and urinary calcium which follow the administration of hydrochlorothiazide, 100 mg twice daily, for 4 days, 42 studies were carried out in 16 normal subjects, 9 patients with hyperparathyroidism, and 7 vitamin D-treated subjects with hypoparathyroidism. In six studies in normal subjects, daily sodium losses during thiazide administration were quantitatively replaced, and in six other studies the effect of equivalent sodium losses produced by furosemide was evaluated. Although the magnitude of sodium losses was similar in three groups during therapy with thiazides, urinary calcium fell and urinary phosphorus increased significantly only in normal subjects and those with hyperparathyroidism; no change occurred in patients with hypoparathyroidism. With the replacement of the thiazide-induced sodium losses by NaCl in normals, urinary calcium did not change as urinary sodium increased 4- to 5-fold. Furosemide administration produced similar sodium losses while urinary calcium remained at or above control levels. After correction for changes in plasma protein concentration caused by thiazide-induced hemoconcentration, mean levels of serum calcium were significantly increased only in subjects with hyperparathyroidism and vitamin D-treated patients with hypoparathyroidism. The results indicate that both depletion of extracellular fluid volume and the presence of the parathyroid glands are necessary for the decrease in urinary calcium in response to thiazide therapy. Both the reduction in urinary calcium and increase in urinary phosphate after the use of thiazides may be due, in part, to potentiation of the action of the parathyroid hormone on the nephron. The rise in serum calcium could be due to thiazide-induced release of calcium from bone into extracellular fluid, particularly in states where bone resorption may be augmented, i.e., vitamin D therapy or hyperparathyroidism.