FK506 promotes adenosine release from endothelial cells via inhibition of adenosine kinase.

The immunosuppressants, cyclosporin A and tacrolimus (FK506) induce an increase in plasma levels of adenosine and mimic ischemic preconditioning. However, the mechanism of action of the two drugs on adenosine metabolism is not clear. Since inhibition of adenosine kinase promotes an increase in endogenous adenosine release, we tested a hypothesis that FK506 induces adenosine release via inhibition of adenosine kinase activity. In cultured endothelial cells, FK506 enhanced release of tracer adenosine and inhibited uptake of tracer adenosine. It also reduced adenosine kinase activity of the cell membrane fraction. In addition, FK506 does not inhibit membrane transport of tracer adenosine. These observations indicate that FK506 inhibits in situ adenosine kinase activity in endothelial cells. Other cell signaling inhibitors were found to inhibit adenosine uptake via inhibition of adenosine transport. In conclusion, FK506 promotes adenosine release from endothelial cells by a novel mechanism involving inhibition of adenosine kinase activity associated with the membrane.

Cellular and molecular actions of adrenomedullin in glomerular mesangial cells.

Adrenomedullin (AM), a potent vasodilatory and hypotensive peptide produces several biological outcomes in glomerular mesangial cells. Mesangial cells are important in the pathogenesis of glomerulonephritis, and therefore the actions of AM on mesangial cells have important clinical and therapeutic implications. This minireview describes the various actions of AM on mesangial cell function and the signal transduction mechanisms involved. As in other systems, most actions of AM can be explained by increase in cAMP levels in the cell, although a few exceptions remain. The fact that most data obtained to date has been in culture, the physiological significance of the actions of AM in mesangial cells is discussed.

Microalbuminuria in urban Zimbabwean women.

The prevalence of microalbuminuria (MAU) in African populations has not been reported, nor has the relationship between MAU and hypertension been reported for these populations. We collected spot urine samples from 370 women, 25 years and older as a part of a population-based, cross-sectional blood pressure survey in an urban community in Zimbabwe and analysed the samples for albumin and beta2-microglobulin. The age-adjusted prevalence of hypertension was 30% for women 25 years and older in this community. After excluding the samples with hematuria (11%), the prevalence of MAU (3.0 < or = albumin-to-creatinine ratio (ACR, mg/mmol) <25.0) in the study population was 9%. When age-adjusted to the population in the community, the prevalence was 8% among women 25 years and older. The prevalence of MAU was substantially higher in hypertensive (HT) than in normotensive (NT) women (16% vs 4%, P<0.001). A significantly higher level of log ACR in HT was found in each age group except the youngest age group (age 25-34). In age-adjusted multiple regression, percent fat mass was negatively associated with log ACR (beta = -1. 18, 95% CI (-0.23, -2.21), P = 0.02). In a similar regression analysis, higher log beta8-microglobulin-to-creatinine ratio was very strongly associated with higher log ACR (beta = 0.34, 95% CI (0.25, 0.43), P<0.0001) and significantly associated with lower percent fat mass (beta = -1.02, 95% CI (-0.25, -1.8), P = 0.01). These results suggest that MAU is frequently caused by hypertension, but that other diseases may contribute to its presence.

Economic development and women's blood pressure: field evidence from rural Mashonaland, Zimbabwe.

A survey of 515 non-pregnant women at 12 geographically chosen research sites in rural Mashonaland shows significant differences in mean blood pressure, controlled by age cohorts. Three levels of economic development are identified: (1) the traditional economy on communal lands, with lowest blood pressure, (2) the wage economy in areas of large-scale commercial agriculture, with elevated blood pressure and (3) the wage economy in mining areas, with the highest elevation of blood pressure. The area is dominated by the primate city, Harare, up to distances of 300 km and beyond, from which forces of change and modernization emanate. It is seen that potassium, sodium and the sodium potassium ratio, are distance-related to Harare and that women's blood pressures tend to follow suit. The rise of body sodium in young persons at risk, often accompanied by declining potassium intake and other changes of modernization, suggest that more attention should be focused on rural areas in Africa, now in the throes of economic change.

Learning the regulation of peripheral blood flow.

Students can learn a great deal about the peripheral circulation when teaching is based on five building blocks: hemodynamic principles, neurohumoral control, and three elements of local control of blood flow (metabolic, myogenic, and paracrine). Study of a particular special circulation starts with the application of these building blocks in the context of the function of that tissue. For example, control of skin blood flow is largely concerned with regulation of body temperature (neurohumoral control) and the response to injury (paracrine control). Regulation of coronary blood flow is almost entirely a matter of meeting the metabolic needs of the myocardium (metabolic control). By mixing and matching the five building blocks and keeping in mind the special functions of a particular tissue, students can master the peripheral circulation efficiently.

Effect of variation in environmental temperature on blood pressure: is it important?

OBJECTIVES: To study the effect of variation in environmental temperature on blood pressure and pulse. DESIGN: Repeated blood pressure and pulse measurements in volunteers on two days when environmental temperatures differed by 10 degrees C. SETTING: Part of an ongoing community based study. SUBJECTS: 25 Black subjects (23 males and two females) volunteered from a population cohort participating in an ongoing longitudinal study examining cardiovascular risk factors in an urban African Black environment. MAIN OUTCOME MEASURE: Mean systolic and diastolic blood pressures and pulse rates. RESULTS: The systolic and diastolic blood pressures were significantly higher when recorded at 15 degrees C than at 25 degrees C, mean difference 32.2 +/- 4.2, p < 0.001 and 19.5 +/- 3.0 p < 0.001) for systolic and diastolic blood pressures respectively. The pulse rate per minute at 15 degrees C was significantly lower than at 25 degrees C (mean difference 11.1 +/- 3.2 p = 0.002). CONCLUSIONS: A decrease in environmental temperature by 10 degrees C appears to increase blood pressure. Awareness of this phenomenon is important, especially when surprisingly high blood pressures are observed during low ambient temperatures, to avoid over diagnosis of hypertension. This phenomenon, together with that already established of the white coat hypertension, may lead to the erroneous diagnosis of hypertension.

Inorganic phosphate as regulator of adenosine formation in isolated guinea pig hearts.

This study evaluated cytosolic P(i) as an independent regulator of cardiac adenosine formation by dissociating changes in P(i) from changes in AMP and ADP. Myocardial high-energy phosphates (HEP), measured by (31)P nuclear magnetic resonance spectroscopy, were depleted acutely by perfusing isolated guinea pig hearts with 2-deoxyglucose (2-DG), and the effects of 2-DG were compared with a norepinephrine infusion producing similar changes in HEP. 2-DG treatment resulted in lower adenosine release (R(ado)) (54 +/- 18 vs. 622 +/- 199 pmol x min(-1) x g(-1)) and P(i) concentration ([P(i)]) (0.5 +/- 0.1 vs. 6.0 +/- 0.9 mM) than norepinephrine despite similar AMP concentration ([AMP]). Chronic phosphocreatine depletion produced by beta-guanidinopropionic acid feeding also reduced R(ado) and P(i) during hypoxia. Replacement of perfusate glucose and pyruvate with acetate increased R(ado) (from 39 +/- 12 to 356 +/- 100 pmol x min(-1) x g(-1)) and [P(i)] (from 2.0 +/- 0.5 to 5.1 +/- 0.6 mM) with no change in cytosolic [AMP]. Adenosine kinase isolated from guinea pig hearts was inhibited by [P(i)] values seen during hypoxia or hypoperfusion. We conclude that cytosolic [P(i)] can be an important regulator of cardiac adenosine formation through inhibition of adenosine kinase.

Comparison of the Omron HEM-713C automated blood pressure monitor with a standard ausculatory method using a mercury manometer.

OBJECTIVES: To compare the Omron HEM-713C automated blood pressure machine with the standard ausculatory method using a mercury manometer. DESIGN: Blood pressures of randomly selected subjects were measured using both the Omron HEM-713C and the mercury manometer. SETTING: Dombotombo surburb in Marondera, Zimbabwe. SUBJECTS: One hundred and sixteen subjects 25 years and above (47 males and 69 females) randomly selected in Marondera. MAIN OUTCOME MEASURE: Systolic blood pressure and diastolic blood pressure. RESULTS: The Omron HEM-713C passed with a grade B for both systolic and diastolic blood pressures when using the British Hypertension Society protocol. It also passed both systolic and diastolic criteria for Association of the Advancement of Medical Instrumentation. CONCLUSION: The Omron HEM-713C compares well with the standard mercury manometer, we therefore recommend its use in both research and clinical applications which require blood pressure measurements.

Prevalence of hypertension among women in rural Zimbabwe: a comparison of pregnant and non-pregnant women.

OBJECTIVES: To determine the prevalence of pregnancy induced hypertension (PIH) in rural Zimbabwe. DESIGNS: A blood pressure survey conducted in three socio-economic strata of rural Zimbabwe, communal lands, commercial farms and mining areas. SETTING: Rural Zimbabwe, Mashonaland West Province. SUBJECTS: 627 pregnant (> 20 weeks gestation) and 483 non pregnant (ages 16 to 45) women. MAIN OUTCOME MEASURE: Prevalence of PIH. RESULTS: PIH was defined in two ways: 1. A combination of > or = 2+ proteinuria and systolic and/or diastolic pressure > or = SD above the mean for all pregnant women (> or = 123/75 mmHg). 2. > or = 1+ proteinuria and blood pressure of 140 and/or 90 mmHg. Eight women met definition 1 and seven met definition 2, giving a prevalence of 0.8pc; 1.3pc and 3.2pc in communal lands, commercial farms and mines respectively. In non-pregnant women the prevalence of hypertension (> or = 160 and/or 95 mmHg) was 3.5pc; 5.4pc and 15.1pc in communal lands, commercial farms and mines respectively. CONCLUSION: PIH is rare in rural Zimbabwe. Hypertension among non-pregnant women of child bearing age is most common in mining communities which have many similarities to urban environments.

Adenosine formation during hypoxia in isolated hearts: effect of adrenergic blockade.

Adrenergic receptor blockade has been reported to decrease cardiac adenosine formation and release during hypoxia. We wished to determine whether this occurs by an improvement in the energy supply/demand ratio. Isolated guinea pig hearts were perfused at a constant pressure of 50 mm Hg. Hypoxia (30% O2) was maintained for 20 min while adenosine release and venous PO2 were measured in the coronary venous effluent. beta-adrenergic blockade with 5 microM atenolol did not change hypoxic adenosine release (Control: 15.6 +/- 2.7, Atenolol: 23.6 +/- 5.7 nmol/g/20 min). Addition of 6 microM phentolamine with atenolol significantly reduced hypoxic adenosine release (4.4 +/- 1.4 nmol/g/20 min, P < 0.05). Atenolol was without hemodynamic effects, but addition of phentolamine reduced left ventricular pressure development, heart rate, and oxygen consumption prior to hypoxia. Atenolol plus phentolamine did not change venous PO2 during hypoxia. Treatment with phenoxybenzamine (1 microM) plus atenolol also reduced adenosine release (7.4 +/- 0.8 nmol/g/20 min). Control experiments and atenolol plus phentolamine experiments were repeated using 31P-NMR to measure high energy phosphates. Adrenergic blockade had no effect on phosphate concentrations during normoxia, but resulted in higher [PCr], lower [P(i)] and higher phosphorylation potentials during hypoxia. Adrenergic blockade also prevented the hypoxia-induced rise in intracellular [H+], [AMP] and [ADP] seen in control hearts. The changes in phosphorylation potential are correlated with similar changes in adenosine release in adrenergically intact hearts. We conclude that the primary effect of adrenergic blockade during hypoxia is a reduction in ATP use due to alpha-receptor blockade. This leads to improved high energy phosphate concentrations during hypoxia and a reduction in adenosine formation.

Blood pressure responses to dietary salt in rural and urban African men.

The purpose of this study was to test the hypothesis that urban Africans exhibit more pressor sensitivity to dietary sodium than do their rural counterparts. We studied short-term sodium sensitivity of nonrandomized representatives of rural and urban population groups in which the overall prevalence of hypertension was 7% and 24% respectively. The rural group (mean age: 39 +/- 3 years) had 20 subjects and the urban group (mean age: 37 +/- 2 years) 21 subjects. Sodium sensitivity was assessed by measuring the blood pressure response to acute dietary restriction and sodium loading. In addition, hormonal responses to the dietary sodium changes were observed. We restricted the study to normotensives. Subjects were studied on baseline diet, after 4 days of a low-salt diet providing 10 mEq sodium per day, and after 4 days of a high-salt diet providing 800 mEq sodium per day. On the high-salt diet, half of the subjects received 100 mEq supplemental potassium per day. Mean arterial pressure in rural and urban subjects increased from 79 +/- 2 to 87 +/- 2 and 82 +/- 2 to 89 +/- 2 mm Hg (+/- SEM) respectively when subjects changed from low to high salt. Pulse pressure of rural and urban subjects also increased with the high-salt diet. Potassium supplementation did not influence the pressor response. Using a 10% increase in mean arterial pressure when going from low-salt to high-salt diets as an index of sodium sensitivity, we found that 33% of urban and 35% of rural subjects were sensitive to sodium. We conclude that the sensitivities of rural and urban normotensive men to the pressor effects of sodium are not different.

Adenosine release and high energy phosphates in intact dog hearts during norepinephrine infusion.

Cardiac adenosine release is thought to depend on the oxygen supply/demand ratio, and this effect may be mediated by changes in high energy phosphate concentrations. Previous studies supporting this hypothesis have been done primarily in isolated hearts. We tested this hypothesis in intact dog hearts. Anesthetized, open-chest dogs were placed in a 4.7-T magnet where 31P nuclear magnetic resonance spectra were acquired via a surface coil over the heart at 2-minute intervals (60 scans, 2-second interpulse delay). Coronary sinus flow was shunted through a flow probe and returned via a jugular vein. After a control period, intracoronary norepinephrine was infused (12 micrograms/min) for 16 minutes and plasma samples were taken every 5 minutes. The phosphocreatine/ATP peak area ratio was used as an index of high energy phosphate changes. During norepinephrine infusion, arterial pressure, heart rate, coronary sinus flow, oxygen consumption, and adenosine release all increased significantly. Adenosine release peaked at 5 minutes but remained elevated after 15 minutes. There was a transient fall in the phosphocreatine/ATP ratio (9.2 +/- 3.1%, p less than 0.05) during the first 7 minutes, but the ratio returned to control levels by 9 minutes. The oxygen supply/consumption ratio increased after 5 minutes of norepinephrine infusion and then returned to control levels. We conclude that during norepinephrine infusion in vivo, persistent adenosine release can occur with only small transient changes in high energy phosphate concentrations and with no decrease in the oxygen supply/demand ratio.

Adenosine formation and energy status in isolated guinea pig hearts perfused with erythrocytes.

The purpose of this study was to examine adenosine release and high-energy phosphate concentrations during norepinephrine (NE) infusion in isolated guinea pig hearts perfused with a physiological salt solution (PSS) containing erythrocytes (RBC). Phosphate concentrations were monitored using 31P-nuclear magnetic resonance spectroscopy while NE was infused at 6 x 10(-10) mol/min. Compared with perfusion with PSS alone, RBC-perfused hearts consumed more oxygen and developed higher left ventricular pressure and first time derivative of left ventricular pressure at lower coronary flow rates. Adenosine release rates were very similar with both perfusates. NE infusion did not produce a decline in ATP concentration ([ATP]) or an increase in calculated [ADP] and [AMP] in RBC-perfused hearts. However, phosphorylation potential ([ATP]/[ADP][Pi]) declined because of increased [Pi]. We conclude that NE infusion does not change adenine nucleotide concentrations in well-oxygenated guinea pig hearts and that changes in nucleotide concentrations are not necessary for increased adenosine release. Phosphorylation potential is a better predictor of adenosine release than any individual nucleotide or phosphate concentration.

Blood pressure response to acute changes in dietary sodium in young Zimbabwean men.

OBJECTIVE: The primary objective of this study was to determine the effect of acute alterations in sodium intake upon the blood pressure and hormone levels of young Zimbabwean men. DESIGN: Blood pressure, 24-h urinary electrolyte excretion and plasma concentrations of angiotensin II, aldosterone, and atrial natriuretic peptide were measured in normotensive black medical students. Three sets of measurements were taken: (1) during free access to sodium (baseline); (2) after 4 days on a low-sodium diet (10 mmol/day); and (3) after 4 days on a high-sodium diet (800 mmol/day). METHODS: Blood pressure was measured by random zero sphygmomanometry, hormone levels by radioimmunoassay, and urinary electrolytes by flame photometry. RESULTS: The low-sodium diet caused the range of pulse pressure to narrow, with a decrease in systolic blood pressure (SBP) and an increase in diastolic blood pressure (DBP). With the introduction of the high-sodium diet, SBP increased and DBP decreased. Mean arterial pressure did not change. At the same time, angiotensin II and aldosterone decreased. Plasma atrial natriuretic peptide did not change. A subgroup of the men on the high-sodium diet also received 100 mmol potassium/day. The increase in SBP associated with high sodium was significantly attenuated by the presence of added potassium. CONCLUSIONS: SBP of young black Zimbabwean men is lowered by dietary sodium restriction and rises with a large increase in dietary sodium for a short duration, but mean arterial pressure does not change due to the opposing decreases in DBP.

Adenosine formation and myocardial energy status during graded hypoxia.

Previous studies using hypoperfusion and 2-deoxyglucose infusion have revealed a biphasic relationship between myocardial energy status and adenosine release (RADO). As energy charge ([ATP] + 1/2[ADP])/([ATP] + [ADP] + [AMP]) or phosphorylation potential ([ATP]/[ADP][Pi]) is lowered there is an initial increase in RADO, but RADO declines from peak levels during severe energy depletion. This study examined the hypothesis that the same pattern of RADO exists during graded hypoxia. Isolated guinea-pig hearts were perfused at constant flow and exposed to mild (30% O2) and severe (0% O2) hypoxia in the presence of norepinephrine (NE, 6 x 10(-8) M). Phosphorylation potential and energy charge were determined using 31P-NMR spectroscopy and adenosine release into coronary venous effluent was measured. Graded hypoxia lowered energy charge and phosphorylation potential, and raised RADO. Although severe hypoxia plus NE lowered energy charge and phosphorylation potential to levels equivalent to those associated with decreased RADO during hypoperfusion or 2-deoxyglucose treatment, RADO during severe hypoxia was greater than during mild hypoxia. HCl was infused during severe hypoxia in order to reproduce the low intracellular pH seen during hypoperfusion, but HCl increased RADO rather than decreasing it. We conclude that during hypoxia, RADO does not have a biphasic relationship to phosphorylation potential or energy charge, suggesting that the regulation of adenosine formation cannot be explained solely in terms of these variables. Furthermore, intracellular acidosis is not responsible for inhibiting RADO at low phosphorylation potential and energy charge during hypoperfusion because it has no effect on RADO during severe hypoxia.

Interstitial adenosine concentration during norepinephrine infusion in isolated guinea pig hearts.

This study determined the effect of norepinephrine (NE) on cardiac interstitial fluid adenosine concentration [( ADO]isf). Isolated guinea pig hearts were perfused with a Krebs-Henseleit buffer solution. Radiolabeled albumin, sucrose, and adenosine were injected under control conditions and after 3 and 20 min of NE infusion to obtain multiple indicator dilution curves that were used to determine capillary transport parameters for adenosine. These parameters together with venous adenosine concentrations were used in a mathematical model to a calculate [ADO]isf. Capillary transport parameters were not changed significantly by NE infusion. Because of uncertainty regarding two model parameters, two sets of [ADO]isf values were calculated. One set used best-fit values obtained from indicator dilution curves, and a second set used parameters chosen to provide the highest [ADO]isf values consistent with indicator dilution curves. Venous adenosine concentrations were 1.9 +/- 0.4 nM under control conditions and 243 +/- 110 and 45 +/- 25 nM after 3 and 20 min of NE infusion, respectively. Calculated [ADO]isf was 2.6-9.4, 591-1,288, and 166-324 nM, respectively, under these same conditions. We conclude that NE infusion greatly increases [ADO]isf, and adenosine is responsible for most of the vasodilation at 3 min. The subsequent fall in venous concentration is due to a fall in [ADO]isf rather than to decreased capillary permeability. Vascular resistance remained low while [ADO]isf fell, which suggests that additional vasodilators are important during maintained NE infusion.

Adenosine formation and energy status during hypoperfusion and 2-deoxyglucose infusion.

The relationship between adenosine (Ado) formation and cytosolic energy status was studied in isolated guinea pig hearts during hypoperfusion plus norepinephrine infusion (0.6 nmol/min) and in isolated rat hearts during 2-deoxyglucose (2-DG) infusion. 31P nuclear magnetic resonance (31P-NMR) was used to measure phosphate concentrations, and both phosphorylation potential (expressed as [ATP]/[ADP][Pi]) and energy charge [expressed as (([ATP] + 1/2[ADP])/([ATP] + [ADP] + [AMP]))] were calculated as indexes of cytosolic energy status. Both progressive flow reductions and increasing length of exposure to 2-DG led to progressive decreases in energy charge and phosphorylation potential. In both cases, steady-state Ado release first increased then declined despite a continued fall in energy status. Inosine release followed a similar pattern. This biphasic pattern of Ado release vs. energy charge is similar to the pattern seen in in vitro studies of cytosolic 5'-nucleotidase, supporting the hypothesis that Ado formation in vivo is regulated by the influence of energy status on this enzyme. However, Ado release in vivo peaked at an energy charge much higher (0.997) than that observed in vitro (0.60-0.86). It is therefore probable that the inhibition of Ado formation in the perfused heart occurs via factor(s) in addition to energy charge.

Dirofilaria immitis: alteration of endothelium-dependent relaxation in the in vivo canine femoral artery.

Pathogenic mechanisms in filarial diseases are complex and poorly understood. While examining endothelium-dependent vasodilatory responses in the in vivo canine femoral artery, we noticed that dogs with Dirofilaria immitis infection had altered vascular responsiveness. The results reported here extend our original observations on vascular reactivity in dogs with D. immitis infection (L. Kaiser, J. F. Williams, E. A. Meade, and H. V. Sparks, 1987, American Journal of Physiology 253, H1325-H1329). In noninfected dogs, acetylcholine binds to the luminal endothelial cell muscarinic receptor. This results in release of a nonprostaglandin endothelium-derived relaxing factor. The relaxing factor causes an increase in vascular smooth muscle guanylate cyclase and relaxation. However, in dogs with D. immitis infection the mechanism of relaxation to acetylcholine is different. At least two endothelium derived relaxing factors are involved: the major factor is a prostaglandin; the second factor works through vascular smooth muscle cGMP. These data suggest that adult D. immitis release pharmacologically active factors that can alter distal endothelial cell function. The notion that filarial products may alter the physiological function of endothelial cells should be considered in the pursuit of improved understanding of pathogenic mechanisms of filariasis.

Transcapillary adenosine transport and interstitial adenosine concentration in guinea pig hearts.

We used the multiple-indicator-dilution technique to observe the capillary transport of adenosine in isolated Krebs-Henseleit-perfused guinea pig hearts. Tracer concentrations of radiolabeled albumin, sucrose, and adenosine were injected into the coronary inflow; outflow samples were collected for 10-25 s and analyzed by high-performance liquid chromatography (HPLC) and by gamma- and beta-counting. The albumin data define the intravascular transport characteristics; the sucrose data define permeation through interendothelial clefts and dilution in interstitial fluid (ISF). Parameters calculated from adenosine data include permeability-surface area products for endothelial cell uptake at the luminal and abluminal membranes and intraendothelial metabolism. We found that in situ endothelial cells avidly take up and metabolize adenosine. Tracer adenosine in the capillary lumen is twice as likely to enter an endothelial cell as it is to permeate the clefts. There was no adenosine in the arterial perfusate. Under control conditions, the steady-state venous adenosine concentration was 3.6 +/- 0.8 nM, which from the flow and the parameters estimated from the tracer data gave a calculated ISF concentration of 6.8 +/- 1.5 nM. During dipyridamole infusion (10 microM) at constant pressure, the cell permeabilities went essentially to zero, whereas the venous adenosine concentration increased to 44.0 +/- 12.6 nM, giving an estimated ISF concentration of 191 +/- 53 nM. With constant flow perfusion, venous concentration during dipyridamole infusion was 30.9 +/- 6.3 nM, and estimated ISF concentration was 88 +/- 20 mM. We conclude that in this preparation, at rest, the ISF adenosine concentration is about twice the venous concentration and the ISF adenosine concentration increases with dipyridamole administration.