Improved high-performance liquid chromatographic method in the analysis of adenovirus particles.

We developed a HPLC method on a novel continuous bed matrix (UNO Q, Bio-Rad) for the direct quantification of adenoviral type 5 (Ad5) particles produced in 293S Human Embryonic Kidney cells and compared this with an existing HPLC method on a conventional ion-exchange resin (Resource Q, Pharmacia). The 293S cell extract contained large amounts of DNA. This contaminated the viral peak on the Resource Q column and only after Benzonase treatment was it possible to quantify the viral particles in the cell extract. In contrast, the virus peak on the UNO Q column was resolved from the DNA which eliminates the need for pretreatment of the sample with Benzonase. Cross-analysis of the Ad5 fraction from the UNO Q column using a size-exclusion HPLC column revealed no additional contaminating peaks. We conclude that the purity of the Ad5 virus peak on the continuous bed matrix UNO Q column was superior to the purity of the virus on the conventional Resource Q column, which is essential for reliable quantification.

Children are different: environmental contaminants and children's health.

Although the impact of environmental contaminants on human health has been widely studied, few reports in the Canadian literature have focussed on the specific vulnerability of children. Because of their rapid growth, physiologic and metabolic immaturity, the fetus and child are often at increased risk from toxic substances in their environments. Furthermore, greater air, food and fluid intakes relative to body weight compared with the adult, increase the child's potential for excessive exposures. The crawling stage of infancy, the play patterns and short stature of toddlers also serve to increase their exposure to dust and heavy and volatile substances which accumulate near the floor. This article provides an overview of some of the developmental physiologic, anatomic and behavioural features of the fetus, infant and child which increase their vulnerability to environmental contaminants in comparison with adults. Specific examples are given.

alpha- and beta-adrenoceptor blockade fail to prevent high sodium diet-induced left ventricular hypertrophy.

High sodium diet (HS, 8% NaCl) induces left ventricular hypertrophy (LVH) in normotensive rats without an increase in pressure or volume load or in resting cardiac sympathetic activity. HS may affect LV adrenoceptors density or affinity or their postreceptor pathways, thereby causing LVH. We therefore assessed the effects of HS with and without blockade of alpha1- or beta-adrenoceptors by terazosin or nadolol, alone or in combination, on resting hemodynamics, LV and right ventricular (RV) weights, and LV dimensions of male WKY rats. HS increased LV weight by 14% to 17%, and the ratio of LV wall thickness to radius by 18% to 23%. Singly or in combination, the adrenoceptor antagonists did not prevent HS-induced LVH, but instead aggravated it. The increased ratio of LV wall thickness to radius and of LV to RV were attenuated by terazosin or nadolol alone. Neither the resting LV peak-systolic or end-diastolic pressures nor the right atrial pressure was changed by HS, either alone or in combination with the blockers. The failure of chronic alpha1- or beta-blockade to prevent HS-induced LVH suggests that adrenoceptor activation is not important in evoking the LVH. However, the blockers shifted the LVH from a concentric to an eccentric form, suggesting an involvement of additional trophic factors during adrenoceptor blockade.

Myocardial kinetics of technetium-99m teboroxime in the presence of postischemic injury, necrosis and low flow reperfusion.

OBJECTIVES: This study evaluated technetium-99m (Tc-99m) teboroxime kinetics in postischemic and partially necrotic myocardium with complete and low flow reperfusion using an isolated perfused rat heart model. BACKGROUND: Technetium-99m teboroxime has been proposed for use in the early diagnosis of reperfusion after thrombolysis on the basis of models of myocardial necrosis with complete reperfusion. Clinically, however, reperfusion is frequently incomplete, resulting in a mixture of necrotic, ischemic and postischemic tissue. METHODS: Hearts were classified into five groups: group 1 (n = 8, control); group 2 (n = 7, 30 min of no flow with complete reperfusion); group 3 (n = 12, 60 min of no flow to induce partial necrosis, followed by complete reperfusion); group 4 (n = 8, continuous low flow without flow interruption); and group 5 (n = 9, 60 min of no flow with low flow reperfusion). Buffer containing Tc-99m teboroxime was perfused for 15 min, followed by tracerfree buffer for 35 min, to evaluate uptake and clearance, respectively. RESULTS: Uptake slopes for groups 1 to 5 were (mean +/- SD) 3.0 +/- 0.7, 2.6 +/- 0.8, 2.1 +/- 0.5, 0.8 +/- 0.2 and 0.8 +/- 0.3, respectively (p < or = 0.0005 for groups 1, 2 and 3 vs. groups 4 and 5, and p = 0.003 for group 3 vs. groups 1 and 2). Clearance curves from groups 1 to 3 were best fit by a biexponential function (p < 0.001); those from groups 4 and 5 were monoexponential. In groups 1, 2 and 3, the initial clearance rate constants (ki) (0.9 +/- 0.5 x 10(-3); 1.0 +/- 0.2 x 10(-3); 1.1 +/- 0.5 x 10(-3) s-1, respectively) and the monoexponential rate constants (Kmono) (2.0 +/- 0.3 x 10(-4); 2.2 +/- 0.4 x 10(-4); 2.1 +/- 0.2 x 10(-4) s-1, respectively) were significantly greater than those in groups 4 and 5 (0.9 +/- 0.5 x 10(-4); 1.2 +/- 0.3 x 10(-4) s-1, respectively, p < or = 0.005). CONCLUSIONS: The uptake and initial clearance kinetics of Tc-99m teboroxime depend mainly on myocardial flow in this model. The presence of partial necrosis and postischemic injury has little effect on the initial clearance but leads to some reduction in uptake at normal flow rates. Evaluation of Tc-99m teboroxime kinetics may permit early noninvasive detection of inadequate reperfusion in acute myocardial infarction.

Differences in distribution of lymphocyte antigens in chicken lines divergently selected for antibody responses to sheep red blood cells.

The proportion of cells showing differentiation antigens specific for T cells, B cells and leukocytes was studied at various ages in peripheral blood, and at 14 weeks of age in the bursa of Fabricius, spleen and thymus of two lines of chicken that had been selected over 13 generations for either high (H) or low (L) antibody responses to sheep red blood cells (SRBC), and also in a randombred control (C) line. Flow cytometry showed no consistently significant differences between the three lines in numbers of circulating lymphocytes and other leukocytes after hatching. However, higher percentages of CD4+ cells and B cells were present in the spleen and thymus from the H line compared with the L line. However, the L line was characterized by a higher proportion of splenic CD8+ cells and spleen cells expressing gamma-delta T-cell receptors. Immunization with sheep red blood cells had no effect on the distribution of CD4+ or CD8+ cells in the various tissues at 2 and 7 days after immunization. These results suggest that previously reported differences in in vivo immune responses between these chicken lines may be related to the differences in resident T-lymphocyte subpopulations in the lymphoid tissues. The involvement of T-cell subsets and non-antigen-specific mechanisms in divergent selection on humoral immune responses in chickens is discussed.

Dietary sodium and Na,K-ATPase activity in Dahl salt-sensitive versus salt-resistant rats.

OBJECTIVE: To investigate the effects of high dietary sodium on brain and kidney Na,K-ATPase activity in Dahl salt-sensitive (Dahl-S) and salt-resistant (Dahl-R) rats. METHODS: From the age of 4 weeks Dahl-S and Dahl-R rats were fed either standard or high-sodium diet (8% sodium chloride) for 3 weeks. The hydrolysis of [gamma-32P]-ATP in the absence or presence of various concentrations of ouabain was used to determine apparent Na,K-ATPase activity and its isoform composition. To assess whether reduced Na,K-ATPase activity and its isoform composition. To assess whether reduced Na,K-ATPase activity was caused by an endogenous inhibitor, brain and kidney microsomes were pre-incubated with antibody Fab fragments (Digibind). RESULTS: The high-sodium diet increased mean arterial pressure in the Dahl-S but not in the Dahl-R rats. Two binding sites (alpha 1 and alpha 2) in several areas of the brain and one binding site in the kidneys (alpha 1) were detected. The high-sodium diet reduced Na,K-ATPase activity in the hypothalamus of the Dahl-S but not of the Dahl-R rats, but did not cause changes in the brain cortex, pons or kidney. The Na,K-ATPase isoform composition in the brain cortex, hypothalamus and pons and kidney was not changed by the high-sodium diet. In the rats fed the standard-sodium diet, Digibind increased Na,K-ATPase activity only in the hypothalamus of the Dahl-S rats. In rats fed the high-sodium diet, Na,K-ATPase activity was increased by Digibind in the hypothalamus of both strains of rats, but by more in the Dahl-S rats. CONCLUSION: The present data indicate that a high-sodium diet inhibits hypothalamic Na,K-ATPase via increased binding of an inhibitor.

Dietary sodium and central vs. peripheral ouabain-like activity in Dahl salt-sensitive vs. salt-resistant rats.

To assess the possible contribution of brain ouabain-like activity (OLA) to the pressor effects of high-sodium intake in Dahl salt-sensitive (Dahl S) rats, we assessed the effects of high (8%) on blood pressure (BP) and peripheral and brain OLA in Dahl on blood pressure (BP) and peripheral and brain OLA in Dahl S and Dahl salt-resistant (Dahl R) rats. On regular sodium intake, Dahl S and R had similar BP; however, by 7 wk of age adrenal and plasma OLA were 15-30% higher in Dahl S vs. R, whereas central OLA remained similar. On high-sodium intake, in Dahl S both peripheral and central OLA increased within 1 wk with additional increases after 3 wk. These increases preceded the rise in BP. In Dahl R rats, high sodium did not increase BP. However, 3 wk of high sodium did increase peripheral as well as central OLA, the latter to a lesser extent compared with Dahl S and not in the hypothalamus. These results are consistent with the concept that central OLA may be involved in the pressor responses to high sodium in Dahl S. Circulating OLA may play a role in the regulation of renal function to excrete excess sodium in both strains.

Stretch-mediated activation of cardiac renin gene.

This study was designed to quantitate cardiac mRNA levels encoding components of the local renin-angiotensin system during the development of volume overload-induced cardiac hypertrophy. Changes in cardiac renin mRNA levels were measured in relation to renin activity in the left ventricle (LV) and in plasma after acute passive stretch of the heart caused by an aortovenocaval shunt in the rat. A quantitative reverse-transcriptase polymerase chain reaction method with competitive internal standards was used to measure mRNA levels in total RNA derived from cardiac tissues after shunt. Seven days after shunt surgery, LV weight was increased by 23%. Renin activities were elevated four- and twofold in plasma and LV, respectively. LV angiotensinogen mRNAs were not significantly increased by shunt surgery; they were twofold higher than phosphoglycerate kinase mRNA from the housekeeping gene PGK-1. By day 7, LV levels for renin mRNA were significantly increased from well below 0.25% to approximately 1% of PGK-1 mRNA. Identity between renin polymerase chain reaction products from kidney and heart cDNAs and absence of "reninlike" amplification products were supported by Southern blotting. Volume overload caused increased expression of the renin gene in the stretched myocardium. This finding is consistent with the concept of a myocardial renin-angiotensin system that can be activated by locally produced renin and contributes to the hypertrophy of cardiac muscle.

Immunocytochemical analysis of mitogen responses of carp (Cyprinus carpio L.) peripheral blood leucocytes.

Phytohaemagglutinin (PHA) and lipopolysaccharide (LPS) responses of surface immunoglobulin-positive (sIg+) and surface immunoglobulin-negative (sIg-) carp peripheral blood leucocytes (PBL) were studied. sIg+ cell-enriched and depleted carp PBL populations (sIg+ and sIg- cell fractions, respectively) were obtained by magnetic cell sorting (MACS) and mitogenic stimulation in vitro was measured by 3H-thymidine incorporation. The mitogen responses of sIg+ and sIg- cells in non-separated carp PBL cultures were analysed by simultaneous detection of incorporated 5-bromo-2'-deoxyuridine (BrdU) and sIg with the fluorescence microscope and flow cytometer. Flow cytometric determination of the percentage of sIg+ cells in combination with absolute cell counting, revealed an increase of sIg+ cells but not of sIg- cells after LPS stimulation while the number of sIg- cells and not of sIg+ cells was enhanced after PHA stimulation. LPS stimulation showed an increased 3H-thymidine incorporation in the sIg- cell fraction compared with non-separated cells and BrdU incorporation was observed in sIg- cells from LPS-stimulated cultures by fluorescence microscopy. However, flow cytometric analysis showed that mainly dull sIg+ cells and not sIg- cells are stimulated by LPS. These dull sIg+ cells were not sorted from sIg- cells with MACS and could apparently not be distinguished from sIg- cells by light microscopy. PHA stimulates sIg- cells and not sIg+ cells as was estimated by all techniques used.

Chronic central versus peripheral ouabain, blood pressure, and sympathetic activity in rats.

To assess whether chronic ouabain administration causes hypertension by increasing sympathetic activity, we recorded arterial blood pressure and heart rate at rest and after ganglionic blockade in conscious Wistar rats following 10 to 14 days of central or peripheral administration of ouabain. Intracerebroventricular or intravenous infusion of ouabain (10 micrograms/d for both) as well as subcutaneous ouabain pellets (releasing 25 micrograms ouabain/d per pellet) increased mean arterial pressure by 20 to 30 mm Hg and heart rate by 40 to 60 beats per minute. Ouabain pellets increased blood pressure and heart rate in a dose-related manner. After 2 weeks of all ouabain treatments, ouabainlike activity in plasma was not changed but increased significantly in hypothalamus and adrenals. Ouabainlike activity in the adrenals was increased more by intravenous than subcutaneous or intracerebroventricular ouabain treatment, but the different treatment modes caused similar increases in the hypothalamus. Concomitant central infusion of antibody Fab fragments against ouabain prevented the ouabain pellet-induced increases in blood pressure and heart rate. Ganglionic blockade by intravenous hexamethonium normalized blood pressure and heart rate in ouabain-treated rats. These data suggest that in normotensive rats exogenous ouabain, regardless of the mode of administration, may act centrally to cause sympathoexcitation and thus hypertension.

Age-related increase in sensitivity for ischemic ATP breakdown in hypertrophic hearts of SHR normalized by enalapril.

We evaluated, firstly, the sensitivity to cardiac ischemic ATP breakdown during the development of hypertension and cardiac hypertrophy in Spontaneously Hypertensive Rats (SHR) v Wistar Kyoto (WKY) controls, and secondly, the effects of short-term (8 days) and prolonged (3 months) antihypertensive treatment with the angiotensin converting enzyme inhibitor enalapril on hypertrophy and sensitivity to global ischemia. In isolated perfused hearts, ischemia was induced by a stepwise lowering of the perfusion pressure and the appearance of the ATP breakdown products (purines) in the coronary effluent was assessed as a measure of ischemia. Hearts from 2.5- and 4-month-old SHR started to release purines at a higher perfusion pressure than hearts of WKY, associated with a higher maximum concentration in the coronary effluent. This increased ischemic ATP breakdown in 2.5- and 4-month-old SHR could be attributed to a decreased flow at a given perfusion pressure, because of a two-fold increase in coronary vascular resistance (CVR). In contrast, the maximal purine concentration in the coronary effluent in hearts of 7-month-old SHR was reduced compared to the younger SHR and only slightly higher than 7-month-old WKY, despite a persistent increase in CVR. Enalapril normalized the blood pressure, but only prolonged treatment, significantly prevented and regressed cardiac hypertrophy, and reduced CVR. Whereas enalapril did not influence ATP breakdown in WKY, in SHR both short- and long-term treatment normalized it to the pattern observed in WKY. We conclude that during the early phase of cardiac hypertrophy the hearts of SHR become more sensitive to ischemic ATP breakdown solely because of an increase in CVR, whereas during the established hypertrophic phase, the hearts appear to adapt metabolically, resulting in normalized purine release. Enalapril normalized the transient increase in sensitivity to ischemic ATP breakdown during the development of hypertension in SHR, independent of effects on cardiac hypertrophy, apparently by improving coronary flow at low perfusion pressures.

Sensitivity to ischaemic ATP breakdown in different models of cardiac hypertrophy in rats.

OBJECTIVE: To evaluate the sensitivity to ischaemia of rat hearts made hypertrophic by pressure overload [two-kidney, one clip (2-K,1C) rats], volume overload (aortocaval arteriovenous shunt), minoxidil or isoproterenol. METHODS: Ischaemia was induced in the isolated perfused hearts by a stepwise lowering of the perfusion pressure; at each step the coronary effluent was assessed for the products of ATP breakdown. RESULTS: Hypertension increased cardiac weight by 35 and 56% after 2.5 and 12 weeks, respectively. Volume overload increased heart weight by 25 and 55% after 1 and 12 weeks, respectively. Minoxidil (for 5 weeks) or isoproterenol (for 1 week) increased cardiac weight by 22 and 16%, respectively. The hearts from 2-K,1C rats started to release ATP catabolites in the coronary effluent at a substantially higher perfusion pressure, and with significantly higher maximal levels, than the control hearts. In contrast, in volume overload cardiac ATP breakdown was similar to that in the controls, and isoproterenol administration caused significantly lower levels of ATP breakdown. At identical flow rates, normalized per gram dry tissue, the purine concentration in the coronary effluent was similar in all of the models of cardiac hypertrophy studied and in the respective controls, and was even lower in the long-term volume-overloaded and isoproterenol-induced hypertrophic hearts. CONCLUSIONS: We conclude that hearts from hypertensive rats are more sensitive to ischaemic ATP breakdown during lowering of perfusion pressure than hearts from volume-overloaded or control rats. This is independent of the duration of the hypertrophic process, and can be explained by a lower coronary flow per gram heart tissue at a given perfusion pressure. This conclusion is strengthened by the observation that hypertrophic hearts from volume-overloaded rats had similar amounts of cardiac hypertrophy to the hearts from the hypertensive rats, without a change in flow, coronary vascular resistance or ischaemic sensitivity, whereas the hearts from isoproterenol-treated rats had lower ischaemic sensitivity and coronary vascular resistance.

Dietary sodium stimulates ouabainlike activity in adrenalectomized spontaneously hypertensive rats.

Both the adrenal glands and the hypothalamus have been proposed to produce compound(s) with ouabainlike activity (OLA). To evaluate the contribution of the adrenal glands, 4-wk-old spontaneously hypertensive rats (SHR) were sham operated or adrenalectomized. The adrenalectomized SHR were given daily injections of corticosterone and aldosterone. Subsequently, rats were randomized to control or high (8%) dietary Na+, and after 2.5 wk, blood pressure and OLA in plasma, hypothalamus, and pituitary were evaluated. Hypertension developed somewhat less in adrenalectomized vs. sham-operated SHR. On control Na+ intake, adrenalectomy caused only minor decreases in circulating and central OLA. Adrenalectomy did not prevent the 50-90% increases in plasma, hypothalamus, and pituitary OLA caused by high Na+ intake for 2.5 wk. These findings are consistent with the concept that, at least in SHR, the central nervous system may represent the major source of both central and peripheral OLA.

Effects of dietary sodium on central and peripheral ouabain-like activity in spontaneously hypertensive rats.

High dietary Na+ intake enhances pressor and sympathoexcitatory responses in spontaneously hypertensive rats (SHR) but not Wistar-Kyoto (WKY) rats. To evaluate the possible contribution of central ouabain-like activity (OLA), brain and peripheral OLA was assessed in SHR vs. WKY rats at 4 wk of age and after 2 and 4 wk of high vs. control Na+ intake started at 4 wk of age. In SHR, hypertension developed with maturation and was exacerbated by high Na+ intake. With control Na+ intake, SHR showed higher OLA at 4, 6, and 8 wk of age in the pituitary and hypothalamus and also by 8 wk in the adrenals and left ventricle but not in plasma. High Na+ intake increased OLA in all tissues examined in both WKY rats and SHR. After 2 wk on high Na+, only OLA in hypothalamus and pituitary was higher in SHR vs. WKY rats; after 4 wk on high Na+, peripheral (i.e., adrenals, left ventricle, and plasma) OLA was also higher. These results indicate that in SHR the development of hypertension is associated early on with increases in central OLA and in a later phase with increases in peripheral OLA as well. High Na+ intake increases OLA in both SHR and WKY rats, but the higher OLA may affect sympathetic activity and blood pressure only in SHR.

The renin-angiotensin system and volume overload-induced cardiac hypertrophy in rats. Effects of angiotensin converting enzyme inhibitor versus angiotensin II receptor blocker.

BACKGROUND: The degree of cardiac hypertrophy is not only load dependent: Among other factors, the renin-angiotensin system may play a role in the regulation of cardiac myocyte growth. METHODS AND RESULTS: To evaluate the role of the renin-angiotensin system in volume overload-induced cardiac hypertrophy, we assessed: 1) the time course of changes in cardiac hemodynamics, cardiac anatomy, and plasma and cardiac renin activity in response to volume overload induced by two sizes of abdominal aortocaval shunt and 2) the effects of chronic treatment with an angiotensin converting enzyme inhibitor (ACEI) versus an angiotensin II receptor blocker on hemodynamics and cardiac hypertrophy. Drug treatment started 3 days before shunt surgery. An increase in left ventricular end-diastolic pressure (LVEDP) and the development of right ventricular (RV) and left ventricular (LV) eccentric hypertrophy in response to volume overload occurred within the first week after induction of the shunt. Plasma renin activity (PRA) and cardiac renin activity peaked shortly after induction of the shunt. During the chronic phase, LVEDP and PRA decreased somewhat but remained significantly elevated up to 7 weeks after shunt surgery. Cardiac renin activity returned toward normal within 4 weeks after surgery. Treatment with the ACEI enalapril caused only a modest decrease in LV internal diameter but did not affect increases in LV and RV weights in response to volume overload despite a major decrease in LVEDP after chronic treatment. In contrast, treatment with the angiotensin II receptor blocker losartan, which had similar effects on cardiac and peripheral hemodynamics, prevented dilation of the LV after 7 days and attenuated the dilation of the LV after 28 days. Moreover, increases in LV and RV weights were significantly attenuated by losartan. CONCLUSIONS: The development of volume overload-induced cardiac hypertrophy is associated with significant increases in PRA and cardiac renin activity shortly after induction of an aortocaval shunt. Whereas the two blockers of the renin-angiotensin system decreased LVEDP to a similar extent, only the angiotensin II receptor blocker blunted the hypertrophic response of the heart to volume overload, which is indicative for other than hemodynamic determinants of the cardiac hypertrophic response. One trophic factor may be cardiac angiotensin II generated via an angiotensin II-forming enzyme resistant to ACEI and possibly activated by cardiac volume overload.

Role of brain ouabain-like activity in the central effects of sodium in rats.

To evaluate whether changes in central ouabain-like activity (OLA) play a role in the effects of dietary sodium on sympathetic outflow and blood pressure (BP), the effects of high vs. control sodium intake on BP, brain OLA, and responsiveness to exogenous ouabain were studied in young spontaneously hypertensive rats (SHRs) vs. Wistar-Kyoto (WKY) rats. At 4 weeks of age, the BP of SHRs was already significantly increased compared with that of WKY rats, and hypertension developed further with maturation. High sodium intake exacerbated the hypertension. Compared with WKY rats, in SHRs OLA in the hypothalamus was already increased at 4 weeks of age. High sodium intake increased brain OLA in both SHRs and WKY rats. However, high dietary sodium decreased pressor and sympathoexcitatory responses to exogenous ouabain only in SHRs. These results may indicate that in WKY rats on control sodium, endogenous OLA already exerts its maximal central effects and a further increase in OLA by high sodium intake does not cause hemodynamic changes, whereas in young SHRs the increase in brain OLA by high sodium intake may play a functional role in the pressor and sympathoexcitatory responses to high sodium.

Inosine--a natural modulator of contractility and myocardial blood flow in the ischemic heart?

The energetic role of inosine (INO) remains controversial. The aim of the present study was first to test whether endogenous INO consumption/production correlates with regional myocardial contractile performance and second to test whether locally increased levels of INO influence contractility and blood flow in severely ischemic myocardium. Fentanyl-anesthetized dogs with implanted sonomicrometry crystals and independently perfused left anterior descending coronary arteries were studied. Two relatively load-independent indexes of regional myocardial contractility derived from left ventricular pressure-segment length loops were used: the regional stroke work-end-diastolic segment length relationship (Wr/L(ed)) and the end-systolic pressure-segment length relationship (Plv/L(es)). Very good correlations between myocardial contractile performance (as measured by the slope of the regional Wr/L(ed) relationship) and endogenous INO consumption/production under both nonischemic and ischemic conditions were found. Ischemia severely depressed contractility, significantly shifting rightward the Wr/L(ed) and Plv/L(es) relationships. INO infused into the left anterior descending bypass, in a concentration of 600 to 800 mumol/L, partially restored contractile performance as evidenced by a significant leftward displacement of both relationships. Wr, measured at a common maximum L(ed), increased significantly by 61 +/- 5%. Border-zone collateral flow (microspheres) increased by 35 +/- 7% within the endocardial segments and by 34 +/- 9% in the epicardial segments, but no increase in flow in the ischemic region was measureable. With the current emphasis on recanalization with thrombolytic therapy and considering the apparent safety of INO, this naturally occurring nucleoside might prove to be a useful adjunctive agent in the treatment of acute myocardial ischemia.

Brain ouabain-like activity and the sympathoexcitatory and pressor effects of central sodium in rats.

Intracerebroventricularly infused hypertonic saline elicits sympathoexcitatory and pressor effects. To clarify the mechanisms mediating these effects, we evaluated blood pressure (BP), heart rate (HR), and renal sympathetic nerve activity (RSNA) responses to intracerebroventricular administration of 0.3 M NaCl, ouabain, and rat hypothalamic and pituitary extracts containing ouabain-like activity (OLA) in conscious Wistar rats, before and after intracerebroventricular preinjection of digoxin-specific antibody Fab (DAF) fragments. To exclude modulatory effects of arginine vasopressin (AVP), treatment with DAF fragments was in all experiments preceded by intravenous injection of the AVP antagonist [beta-mercapto-beta,beta-cyclopentamethylenepropionyl1,o- Me-Tyr2,Arg8]AVP. After AVP antagonist pretreatment, 0.3 M NaCl i.c.v. at 3.8 microliters/min for 10 minutes caused simultaneous increases in BP, RSNA, and HR. After AVP antagonist pretreatment, intracerebroventricular injections of 0.3 and 1.0 microgram/l microliter ouabain or the OLA equivalent to 1 microgram ouabain/2 microliters elicited similar significant increases in BP, HR, and RSNA. After pretreatment with AVP antagonist and DAF fragments (66 micrograms/4 microliters i.c.v.), BP, HR, and RSNA responses to 0.3 M NaCl, ouabain, and OLA were all significantly diminished. In contrast, combined AVP blockade and DAF fragments did not affect the BP response to intracerebroventricular angiotensin II, the BP, HR, and RSNA response to intracerebroventricular carbachol and to air stress, or the HR and RSNA responses to intravenous sodium nitroprusside. Intracerebroventricularly injected gamma-globulins (66 micrograms/4 microliters) did not affect the responses to 0.3 M NaCl, ouabain, or OLA.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary sodium induced cardiac hypertrophy.

In humans, high sodium intake not only increases the blood pressure, and thus can cause left ventricular hypertrophy (LVH), but also appears to increase LVH independent of this increase in blood pressure. In both normo- and hyper-tensive rats the hypertrophic effect of increased dietary sodium intake on the heart has been clearly established. In normotensive rats, this effect is strain and age dependent, and seems independent of hemodynamic effects of high sodium intake. In both rats and humans, dietary sodium appears to increase wall thickness, resembling pressure overload rather than an increased left ventricular diameter as expected of volume overload. The mechanisms through which high dietary sodium induces hypertrophy are still unknown. It is possible that dietary sodium increases either adrenergic stimulation and (or) enhances sensitivity for adrenergic stimulation and that this hypertrophic response mainly acts via stimulation of alpha 1-adrenergic receptors. Stimulation of the alpha 1-adrenergic receptors will increase the inositol phosphate-diacyl glycerol pathway and enhance the Na+/H+ exchange. The activity of this exchanger might play an important role in the development of dietary sodium induced cardiac hypertrophy.

Cyclosporine-associated reduction in systolic myocardial function in the rat.

The cardiotoxic effect of cyclosporine in the inbred rat was investigated in the isolated perfused heart model. Diastolic and systolic function was studied in 5 groups of (Lewis x Brown Norway) F1 rats treated with different doses of cyclosporine for different time intervals and in a group which received the vehiculum cremophor. Of the cyclosporine-treated rats, one group was studied four weeks after drug discontinuation to assess reversibility of the effect. To control for the nephrotoxic side effects of cyclosporine, a group of animals that had undergone 1.5-1.75 renal ablation was included. Left ventricular pressures were measured with a balloon catheter; stepwise increases in preload were obtained by small increments in the balloon volume. Over the range of left ventricular volumes studied, no differences were found in diastolic pressure between the groups. Peak systolic pressures were significantly lower in hearts obtained from cyclosporine treated rats which had received 15 or 7.5 mg/kg/24 h subcutaneously for three weeks (P less than 0.05). Our results indicate that cyclosporine treatment resulted in a reversible decrease in myocardial contractile force apparently unrelated to decreases in renal function or to changes in mean arterial pressure.