Slow platelet recovery after PBPC transplantation from unrelated donors.

The effects of the composition of PBPC grafts from matched related donors (MRDs) and matched unrelated donors (MUDs) have not been compared. In a single-center study, the compositions of 55 MRD PBPC grafts and 33 MUD grafts were studied for their effect on the rate of engraftment in patients who had evidence of donor cell engraftment on day +28. The MUD grafts came more frequently from young male donors and contained more CD34(+) cells but similar numbers of colony-forming units granulocyte-macrophage (CFU-GM) and burst forming units-erythroid. The recovery of neutrophils to >500/mm(3) was equally fast in both groups, but recovery of platelets to >20,000/mm(3) was significantly delayed in the MUD group (P<0.001). The MUD group also required more transfusions of platelets and red cells. Patients receiving grafts containing low numbers of CFU-GM had markedly delayed platelet recovery. The patients with the slowest engraftment tended to have prolonged transportation times. Storage experiments suggested a major loss of viable CD34(+) cells and CFU-GM when undiluted PBPC products are stored at room temperature. The data suggest that a fraction of the MUD grafts suffer during transportation. In vitro proliferation assays should be part of the validation and auditing of transportation of MUD grafts.

Protein prenylation is required for aldosterone-stimulated Na+ transport.

Aldosterone stimulation of transcellular Na+ flux in polarized epithelial cells is dependent on at least one transmethylation reaction, but the substrate of this signaling step is unknown. Because it is clear that the majority of cellular protein methylation occurs in conjunction with protein prenylation, we examined the importance of prenylation to aldosterone-stimulated Na+ transport in the A6 cell line. Lovastatin, an inhibitor of the first committed step of the mevalonate pathway, inhibits the natriferic effect of aldosterone but does not inhibit insulin-stimulated Na+ flux. The addition of a farnesyl group does not appear to be involved in aldosterone's action. Neither alpha-hydroxyfarne-sylphosphonic acid, an inhibitor of farnesyl:protein transferase, nor N-acetyl-S-farnesyl-L-cysteine, an inhibitor of farnesylated protein methylation, inhibits the hormone-induced increase in Na+ transport. In contrast, N-acetyl-S-geranyl-geranyl-L-cysteine, an inhibitor of geranylgeranyl protein methylation, completely abolishes the aldosterone-induced increase in Na+ flux with no effect on insulin-mediated Na+ transport or cellular protein content. These data indicate that methylation of a geranylgeranylated protein is involved in aldosterone's natriferic action.

Recruitment strategies for black women at risk for noninsulin-dependent diabetes mellitus into exercise protocols: a qualitative assessment.

The literature is devoid of any specific data describing exercise therapy in blacks at risk for diabetes. The increasing and striking prevalence of obesity and diabetes among several indigenous populations demonstrates the unfortunate interplay between genetic predisposition and a "modern" sedentary lifestyle. Any successful intervention to reduce the risk of acquiring or attenuating the severity of diabetes must focus on behavioral, cultural, psychosocial, and social factors that are amenable to change. Thus, the objective of this study is to present qualitative data that can be useful in the recruitment of blacks into exercise protocols that could prove to be beneficial in preventing diabetes. Focus groups were conducted on 57 black women residing in Washington, DC, Columbia, Maryland, and Hartford, Connecticut. Barriers to exercising included lack of child care, lack of transportation, neighborhood constraints, and family. Incentives that would increase black women's ability to participate in an exercise protocols include transportation, child care, and an exercise environment that includes blacks.

Tachycardic and hypertensive effects of centrally administered clonidine in conscious rats.

The centrally mediated cardiovascular changes induced by clonidine were studied in conscious rats. Clonidine administered intracerebroventricularly (i.c.v.), and intravenously (i.v.) caused hypotension following an initial pressor response. I.v. clonidine caused significant greater hypotension than i.c.v. clonidine (30 micrograms/kg; p less than 0.05). With the 30 micrograms/kg i.c.v. dose, a tachycardia was observed in all rats following initial transient bradycardia. No tachycardia was observed when clonidine was administered i.v. Propranolol (3 mg/kg i.v.) did not modify the cardiovascular actions of i.c.v. clonidine except initial pressure response. While combined treatment with propranolol (3 mg/kg i.v.) and atropine (1 mg/kg i.v.) abolished both the bradycardic and tachycardic actions of i.c.v. clonidine (30 micrograms/kg), but did not modulate the hypotensive action. Yohimbine (30 micrograms/kg i.c.v.) converted the hypotension induced by i.c.v. clonidine (30 micrograms/kg) to hypertension, attenuated the bradycardia but did not modulate the tachycardia. The same dose of i.c.v. yohimbine attenuated the hypotensive effect of i.v. clonidine (30 micrograms/kg) but did not affect the initial pressor response. Prazosin (30 micrograms/kg i.c.v.) did not modulate either phase of the heart rate response to i.c.v. clonidine. These results provide evidence of centrally mediated pressor and tachycardic actions of clonidine in conscious rats. The tachycardia appears to be mediated through the inhibition of parasympathetic tone and is not dependent on alpha-adrenoceptor mechanism. In conscious rats the opposing influence of centrally mediated pressor and depressor actions may result in the apparently low hypotensive potency of i.c.v. clonidine.

The contribution of vasopressin and angiotensin to the maintenance of blood pressure after autonomic blockade.

The contribution of vasopressin and angiotensin II to the maintenance of blood pressure after short-term autonomic blockade was investigated in conscious Long-Evans and Brattleboro (vasopressin-deficient; hereditary diabetes insipidus) rats. After short-term autonomic blockade by atropine (1 mg/kg), propranolol (5 mg/kg), and pentolinium (5 mg/kg and 10 mg/kg/hr), the fall in blood pressure was significantly greater in Brattleboro rats than in Long-Evans rats (48 +/- 3 vs 32 +/- 2 mm Hg; p less than 0.01). Administration of the vasopressin vascular receptor antagonist D(CH2)5Tyr-(Me)AVP (2 micrograms/kg) caused further blood pressure decreases only in Long-Evans rats, so that the final blood pressure in both groups was identical. Administration of enalaprilat (10 mg/kg), an angiotensin converting enzyme inhibitor, further reduced blood pressure in both strains. When enalaprilat was given first after autonomic blockade, it reduced blood pressure in Brattleboro rats but not in Long-Evans rats. Administration of the vasopressin antagonist after enalaprilat further reduced blood pressure only in Long-Evans rats. The fall in blood pressure following vasopressin blockade was greater than that occurring after angiotensin converting enzyme inhibition (14 +/- 1 vs 6 +/- 1 mm Hg; p less than 0.05) in autonomic blockade Long-Evans rats. Plasma levels of vasopressin in Long-Evans rats increased markedly after short-term autonomic blockade, whereas plasma renin and angiotensin II levels were unchanged. Plasma angiotensin II levels were increased by the vasopressin antagonist and decreased by enalaprilat. We conclude that, due to sympathetic nervous system blockade and consequent blunting of renal renin release, vasopressin has a greater capacity than the renin-angiotensin system for maintaining blood pressure after short-term autonomic blockade.

Endogenous vasopressin modulates the baroreflex sensitivity in rats.

The effect of endogenous vasopressin on the baroreceptor reflex has been examined by comparing baroreflex function in Brattleboro rats with hereditary hypothalamic diabetes insipidus (DI rat) with that in Long-Evans rats (LE rat). Baroreflex function was assessed in conscious unrestrained rats during increases in blood pressure with phenylephrine. The slope of the baroreflex function line in LE rats [(19.0, s.e.m. = 1.4) X 10(-4), n = 34] was significantly steeper than that in DI rats [(6.9, s.e.m. = 0.6) X 10(-4), n = 44, P less than 0.0001]. A subpressor infusion of arginine8-vasopressin (2 ng/kg per min for 2 h i.v.) and an equidose of DDAVP caused bradycardia and increased the baroreflex function slope significantly. Acute volume expansion in DI rat did not change the baroreflex sensitivity. A specific vasopressin vascular receptor antagonist, d(Ch2)5Tyr(Me)AVP, did not alter the baroreflex sensitivity in LE rats. These results suggest that endogenous vasopressin is an important physiological regulator of the baroreflex sensitivity in normal rats.

The vascular interaction of noradrenaline and 5-hydroxytryptamine.

The action of 5-hydroxytryptamine (5-HT) to potentiate vasoconstrictor responses to noradrenaline has been studied using the rat isolated perfused mesenteric artery. Receptor antagonists have been employed to compare the vascular receptors which mediate the vasoconstrictor and potentiating actions of 5-HT. In preparations pretreated with methysergide (1 mumol X 1(-1)), cyproheptadine (1 mumol X 1(-1)) or fluoxetine (1 mumol X 1(-1)) the vasoconstrictor response to 5-HT (1 mumol X 1(-1)) was abolished, while ketanserin (1 nmol X 1(-1)) antagonized the response by 90%. In contrast, only methysergide antagonized the sensitizing effect of 5-HT, while the other three antagonists exerted no effect on this action. In view of the different sensitivities of the vasoconstrictor and potentiating effects of 5-HT to antagonist drugs, it is suggested that the vascular receptors which mediate these actions are different.

Inhibition of renin secretion by clonidine after alpha-adrenoceptor blockade in anesthetized dogs.

Dogs anaesthetised with pentobarbital were used to study the effects of the alpha-adrenoceptor antagonists phentolamine and piperoxan on the clonidine-induced suppression of plasma renin activity (PRA). Given alone, clonidine (30 microgram/kg, i.v.) produced an initial rise in mean arterial pressure (MAP) which was followed by a hypotensive response. These changes in blood pressure were accompanied by decreased in PRA and heart rate (HR). Pretreatment with phentolamine, 1 mg/kg i.v. or 3.3 microgram/kg/min infused into the third cerebral ventricle, or piperoxan, 20 microgram/kg stat. + 5 microgram/kg/min infused either i.v. or intraventricularly, did not modify the clonidine-induced falls in PRA, MAP or HR. All pretreatment regimes, with the exception of intraventricular phentolamine, virtually abolished the initial pressor response to clonidine. These results demonstrate that the renin-lowering action of clonidine in the dog is not inhibited by two classical alpha-adrenoceptor antagonists.

Suppression of vasopressin secretion by clonidine: effect of alpha-adrenoceptor antagonists.

Studies were performed in anesthetized dogs to determine if the diuretic effect of clonidine results from inhibition of vasopressin secretion. Intravenous clonidine (30 microgram/kg) decreased plasma vasopressin concentration (as measured by RIA) from 10.9 +/- 1.5 to 5.0 +/- 1.1 ng/ml (P less than 0.01) in association with a transient increase in arterial blood pressure and a decrease in heart rate. Intravenous administration of two alpha-adrenoceptor antagonists, piperoxane and phentolamine, virtually abolished the pressor effect of clonidine but did not prevent the suppression of plasma vasopressin concentration. Clonidine decreased plasma vasopressin concentration from 11.9 +/- 3.1 to 3.3 +/- 1.0 pg/ml in the phentolamine-treated dogs (P less than 0.01) and from 18.1 +/- 4.5 to 12.4 +/- 3.6 pg/ml in the piperoxane-treated dogs (P less than 0.05). These results provide direct evidence that the diuretic effect of clonidine results from inhibition of the secretion of vasopressin. This inhibition does not appear to be a consequence of the pressor effect of the drug but may result from a direct action in the central nervous system.

The effects of sympatholytic drugs on the cardiovascular response to tilting in anaesthetized cats.

1. Using cats anaesthetized with chloralose and urethane, comparison was made of the abilities of several antihypertensive and sympatholytic drugs to lower systemic blood pressure, and to depress the compensatory cardiovascular responses to bilateral carotid occlusion and to 45 degrees head-up tilting. Similar comparisons were also made of the effects of these drugs on the perfusion pressure of the vascularly isolated autoperfused hindquarters, and the response of this to carotid occlusion and tilting. The effects of bilateral vagotomy and haemorrhage on these responses were also studied. 2. It was found that hypotensive doses of both bretylium and guanethidine (3.0 mg/kg, i.v.) markedly depressed the ability of cats to restore their systemic blood pressure and to constrict their hindquarters vasculature during tilting. Both drugs depressed the carotid occlusion reflex in the systemic, but not in the hindquarters, circulation. Neither propranolol, 2.0 mg/kg, i.v., nor bilaterial vagotomy had any effect on these parameters and haemorrhage sufficient to cause marked hypotension was without effect on the systemic responses to carotid occlusion or tilting. 3. Clonidine (1.0, 5.0 and 25 microgram/kg, i.v.), xylazine (62.5, 125 and 250 microgram/kg, i.v.) and reserpine (0.5 and 2.0 mg/kg, i.v.) all caused considerable hypotension but had no effect on the response to tilting of the systemic circulation, apart from somewhat prolonging recovering time. The highest dose of clonidine moderately depressed the hindquarters perfusion pressure, and the response of this to tilting. 4. Clonidine (5.0 and 25 microgram/kg, i.v.) and xylazine (125 and 250 microgram/kg, i.v.) depressed the systemic pressor responses elicited by the ganglion stimulants DMPP and McN-A-343. This may indicate that the ability of clonidine to prolong the pressure recovery during tilt may be due to impaired peripheral sympathetic transmission. 5. It is concluded that drugs which significantly reduce the compensatory pressure reponses to tilting in anaesthetized cats may also cause postural disturbances in man, whilst drugs which merely prolong the period required for pressure compensation seem much less likely to cause serious clinical impairment of orthostatic reflexes. It appears that the cardiovascular response to bilateral carotid occlusion may not provide a good index of the integrity of orthostatic reflexes.

L-Tryptophan-induced depression of the pressor response to clinidine in anaesthetized rats.

1. The interaction of serotonin precursor L-tryptophan with the pressor responses of the anaesthetized rat to the intravenous injection of clonidine, adrenaline and angiotensin has been studied. 2. Pretreatment of rats with L-tryptophan (100 mg/kg) depressed the pressor response to clonidine but had no effect on the responses elicited by adrenaline or angiotensin. 3. The L-tryptophan-induced depression of the clonidine response was prevented by pretreating rats with either Rö 4-4602, carbidopa, BW 172C58, methysergide or by pithing. 4. Intravenous infusions of serotonin depressed the pressor responses to clonidine, adrenaline and angiotensin in both intact anaesthetized and pithed rats. 5. It is concluded that the depressant action of L-tryptophan is dependent on its conversion within the periphery to serotonin. This action is also dependent on or mediated by the sympathetic nervous system.

Mechanism of suppression of renin secretion by clonidine in the dog.

The mechanism by which clonidine suppresses renin secretion was investigated in pentobarbital-anesthetized dogs in which renal perfusion pressure was controlled by means of an aortic clamp. Clonidine (30 microgram/kg, iv) lowered mean arterial pressure (MAP) from 124 +/- 8 to 104 +/- 4 mm Hg (P less than 0.01) and reduced plasma renin activity (PRA) to 32 +/- 4 percent of the control value (P less than 0.01) after 60 minutes. Ganglion blockade with pentolinium (3 mg/kg, im) decreased MAP from 148+/- 7 to 117 +/- 3 mm Hg (P less than 0.01) and reduced PRA to 55 +/- 13 percent of the control value (P less than 0.05) after 45 minutes. Pentolinium converted the hypotension produced by clonidine to hypertension (108 +/- 9 to 146 +/- 10 mm Hg at 60 minutes, P less than 0.05) and abolished the suppression of PRA (105 +/- 14 percent of control at 60 minutes, P less than 0.05). In a further series of experiments, the effects of oxymetazoline, an alpha-adrenergic receptor agonist which is closely related to clonidine but which does not cross the blood brain barrier, were studied. Oxymetazoline (10 microgram/kg, iv) increased MAP from 127 +/- 3 to 154 +/- 2 mm Hg (P less than 0.01) and elevated PRA TO 176 +/- 22 percent of the control value (P less than 0.02) after 30 minutes. A higher dose of oxymetazoline (30 microgram/kg) increased MAP from 129 +/- 10 to 161 +/- 9 mm Hg (P less than 0.05) and increased PRA to 256+/- 37 percent of control (P less than 0.05) after 30 minutes. Taken together, these data support the hypothesis that the inhibition of renin secretion by clonidine results from a centrally mediated decrease in sympathetic neural activity.

The effects of serotonin precursors on the pressor response to intravenous clonidine in conscious rats.

1. The interaction of the biosynthetic precursors of serotonin with the alpha-adrenoceptor-mediated pressor response to intravenous clonidine was investigated in unanaesthetized rats. 2. Pretreatment with 100 mg/kg of either L-tryptophan or 5-hydroxytryptophan (5-HTP) reduced the magnitude of the pressor response elicited by intravenous clonidine (25 microgram/kg) to 15% and 11%, respectively, of that observed in control rats. 3. The depression by L-tryptophan of the clonidine-induced pressor response could be prevented by pretreatment with either the L-aromatic amino acid decarboxylase inhibitor Rö44602 or the serotonin antagonist methysergide.