Interstitial Transudate Research Articles

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart.

The present protocol describes a unique approach that enables the collection of cardiac transudate (CT) from the isolated, saline-perfused rat heart. After isolation and retrograde perfusion of the heart according to the Langendorff technique, the heart is inverted into an upside-down position and is mechanically stabilized by a balloon catheter inserted into the left ventricle. Then, a thin latex cap - previously cast to match the average size of the rat heart - is placed over the epicardial surface. The outlet of the latex cap is connected to silicon tubing, with the distal opening 10 cm below the base level of the heart, creating slight suction. CT continuously produced on the epicardial surface is collected in ice-cooled vials for further analysis. The rate of CT formation ranged from 17 to 147 µL/min (n = 14) in control and infarcted hearts, which represents 0.1-1% of the coronary venous effluent perfusate. Proteomic analysis and high performance liquid chromatography (HPLC) revealed that the collected CT contains a wide spectrum of proteins and purinergic metabolites.

Renin, Prorenin and the Putative (Pro)renin Receptor

Renin is an aspartic protease that consists of 2 homologous lobes. The cleft in between contains the active site with 2 catalytic aspartic residues.1 Unlike other aspartic proteases such as pepsin or cathepsin D, renin is monospecific and only cleaves angiotensinogen, to generate angiotensin (Ang) I. Ang I is the precursor of the active end-product of the renin-angiotensin system (RAS), Ang II. Renin has also been called active renin to underline that an enzymatically inactive form of renin exists. In 1971, Lumbers found that amniotic fluid, left at low pH in the cold, acquired renin activity.2 Later, Skinner described a similar phenomenon in plasma.3 Acidification was not strictly necessary for this increase in Ang I-generating activity, because incubation at low temperature also increased renin activity, albeit to only 15% of activity after acidification. Soon it was postulated that this inactive, but activatable “big” renin (its molecular weight was 5 kDa higher than that of renin) was the biosynthetic precursor of renin. Hence, it was named prorenin. Only with the cloning of the renin gene in 1984 was prorenin definitively proved to be the precursor of renin.4 For reasons that are unknown, prorenin circulates in human plasma in excess to renin, sometimes at concentrations that are 100-times higher.5 Prorenin has also been demonstrated in plasma of cat, dog, cattle, pig, horse, sheep, rabbit, rat, and mouse. A 43-amino acid N-terminal propeptide explains the absence of enzymatic activity of prorenin. This propeptide covers the enzymatic cleft and obstructs access of angiotensinogen to the active site of renin. (Pro)renin is synthesized as a preprohormone. It contains a signal peptide that directs the protein to the endoplasmic reticulum and ultimately to the exterior of the cell. Both renin and prorenin can be fractionated into multiple species by isoelectric focusing. …

Vascular transfer of adenovirus is augmented by nitric oxide in the rat heart.

Reversible opening of the endothelial barrier remains a major obstacle when hearts are transfected via the coronary system. Our aim was to establish an experimental system permitting the continuous analysis of vascular transfer of virus in the intact heart. Isolated saline-perfused rat hearts were inverted and covered with a latex cap to collect interstitial transudate (IT) on the pericardial surface. Adenovirus (10(9) pfu/ml) was stably labeled with rhodamine fluorescent dye. Analysis of IT and coronary perfusate revealed that under baseline conditions, adenovirus in the IT reached 75% of its vascular concentration within 3 min. The nitric oxide-donors S-nitroso-N-acetyl penicillamine (SNAP) and bradykinin (BK) were the most effective substances to increase total IT volume and adenoviral interstitial concentration. Perfusion with 9% serum markedly reduced IT volume flow and delayed the SNAP/BK effect. Our findings demonstrate that SNAP and BK effectively increased coronary transfer of adenovirus suggesting that the inverted isolated heart is a suitable model to optimize vascular transfer of virus under standardized conditions.

Cardiac endothelin and big endothelin in right-heart hypertrophy due to monocrotaline-induced pulmonary hypertension in rat.

Recent observations suggest the existence of a myocardial endothelin (ET) system and its possible involvement in left-ventricular myocardial hypertrophy and failure. However, nothing is known about the role of myocardial ET in right-ventricular hypertrophy. Rats (80-100 g) were given an intraperitoneal injection of saline (controls) or monocrotaline (50 mg/kg) resulting in pulmonary hypertension-induced myocardial hypertrophy (n = 11 in both groups). After 10 weeks, the animals were sacrificed and hearts perfused in vitro to determine levels of big ET-1 and ET-1 in coronary effluent, interstitial fluid and ventricular tissue homogenates; plasma levels were also determined. In monocrotaline-treated animals, weights of right ventricles were 1.5 and of right atria 1.8-fold higher than in controls (p < 0.05), indicating substantial right-ventricular hypertrophy as also evident from greatly increased myocardial production of atrial natriuretic peptide. Left-ventricular weights were not different. Release of big ET-1 in coronary effluent, and of ET-1 in coronary effluent and interstitial transudate were similar in control and hypertrophic hearts (p > 0.05). Disruption of endothelium with collagenase reduced release of both peptides close to zero, indicating endothelial (not myocardial) origin of the peptides. Levels of big ET-1 and ET-1 were similar in left ventricles of both experimental groups, but lower in right ventricles of hypertrophic than control hearts (p < 0.05), reflecting increased tissue mass rather than reduced peptide production. On the other hand, plasma levels of both peptides and of ANP were twofold and levels of angiotensin II 1.3-fold higher in rats with right-heart hypertrophy than in controls (p < 0.05 in each case). These data do not support a role for local cardiac ET-1 and/or big ET-1 in right-ventricular hypertrophy, but point to blood-borne endothelins as possible mediators.

Localization and production of angiotensin II in the isolated perfused rat heart.

We used a modification of the isolated perfused rat heart, in which coronary effluent and interstitial transudate were separately collected, to investigate the localization and production of angiotensin II (Ang II) in the heart. During combined renin (0.7 to 1.5 pmol Ang I/mL per minute) and angiotensinogen (6 to 12 pmol/mL) perfusion (4 to 8 mL/min) for 60 minutes (n=3), the steady-state levels of Ang II in interstitial transudate in two consecutive 10-minute periods were 4.3+/-1.5 and 3.6+/-1.5 fmol/mL compared with 1.1+/-0.4 and 1.1+/-0.6 fmol/mL in coronary effluent (mean+/-half range). During perfusion with Ang II (n=5), steady-state Ang II in interstitial transudate was 32+/-19% of arterial Ang II compared with 65+/-16% in coronary effluent (mean+/-SD, P<.02). During perfusion with Ang I (n=5), Ang II in interstitial transudate was 5.1+/-0.6% of arterial Ang I compared with 2.2+/-0.3% in coronary effluent (P<.05). The tissue concentration of Ang II in the combined renin/angiotensinogen perfusions (per gram) was as high as the concentration in interstitial transudate (per milliliter). Addition of losartan (10(-6) mol/L) to the renin/angiotensinogen perfusion (n=3) had no significant effect on the tissue level of Ang II, whereas losartan in the perfusions with Ang I (n=5) or Ang II (n=5) decreased tissue Ang II to undetectably low levels. The results indicate that the heart is capable of producing Ang II and that this can lead to higher levels in tissue than in blood plasma. Cardiac Ang II does not appear to be restricted to the extracellular fluid. This is in part due to AT1-receptor-mediated cellular uptake of extracellular Ang II, but our results also raise the possibility of intracellular Ang II production.

Relation between enzyme release and irreversible cell injury of the heart under the influence of cytoskeleton modulating agents

The effects of agents modulating the cytoskeleton, taxol (microtubuli stabilizing), vinblastine (microtubuli destabilizing) and cytochalasin D (actin destabilizing) (10 −6 M each) on enzyme and ATP release as well as on irreversible cell injury were investigated in isolated perfused hypoxic and reoxygenated rat hearts. Enzyme (creatine kinase (CK)) and ATP concentration were assayed in the interstitial transudate and venous effluent. Irreversible cell injury was determined from trypan blue uptake and nuclear staining (NS) of cardiomyocytes in histologic sections. ATP release from nonneuronal cells was only detectable in the interstitial transudate and was not significantly altered by the agents. In controls total CK release (about 4% of total CK) exceeded the percentage of irreversibly injured cells by a factor of 8. Taxol and cytochalasin D abolished the hypoxia/reoxygenation induced interstitial CK release and reduced total CK release to a highly significant extent. The percentage of irreversible injured cells was even more diminished by these agents resulting in a ratio of CK/NS of 40. The effect of cytochalasin D apparently is the consequence of decreased contractile performance as shown by analogous depression by butonedione monoxine (BDM), whereas contractile activity was not altered by taxol. Vinblastine had no influence on CK release but increased the number of irreversibly injured cells significantly. In conclusion, cytoskeletal elements apparently participate in the hypoxia/reoxygenation induced process of release of cytosolic enzymes (CK) and irreversible injury in a different way and extent. Taxol exhibits a cytoprotective effect in isolated perfused rat hearts as evaluated by the extent of enzyme release and irreversible cell injury.

Renin-angiotensin system components in the interstitial fluid of the isolated perfused rat heart. Local production of angiotensin I.

We used a modification of the isolated perfused rat heart, in which coronary effluent and interstitial transudate were separately collected, to investigate the uptake and clearance of exogenous renin, angiotensinogen, and angiotensin I (Ang I) as well as the cardiac production of Ang I. The levels of these compounds in interstitial transudate were considered to be representative of the levels in the cardiac interstitial fluid. During perfusion with renin or angiotensinogen, the steady-state levels (mean +/- SD) in interstitial transudate were 64 +/- 34% (P < .05 for difference from the arterial level, n = 8) and 108 +/- 42% (n = 6) of the arterial level, respectively; the levels in coronary effluent were not significantly different from those in interstitial transudate. Ang I was not detectable in interstitial transudate during perfusion with Tyrode's buffer or angiotensinogen. It was very low in interstitial transudate during perfusion with renin and rose to much higher levels during combined renin and angiotensinogen perfusion. The total production rate of Ang I present in interstitial fluid could be largely explained by the renin-angiotensinogen reaction in the fluid phase of the interstitial compartment. In contrast, the total production rate of Ang I present in coronary effluent and the net ejection rate of Ang I via coronary effluent were, respectively, 4.6 +/- 2.2 and 2.8 +/- 1.3 (P < .01 and P < .05 for difference from 1.0, n = 6) times higher than could be explained by Ang I formation in the fluid phase of the intravascular compartment. Ang I from the interstitial fluid contributed little to the Ang I in the intravascular fluid and vice versa. These data reveal two tissue sites of Ang I production, ie, the interstitial fluid and a site closer to the blood compartment, possibly vascular surface-bound renin. There was no evidence that the release of locally produced Ang I into coronary effluent and interstitial transudate occurred independently of blood-derived renin or angiotensinogen.

Interstitial noradrenaline concentration of rat hearts as influenced by cellular catecholamine uptake mechanisms.

Marked concentration differences of noradrenaline (NA) between the vascular and the interstitial compartment were detected by sampling interstitial transudate from isolated perfused rat hearts. The ratios of vascular/interstitial concentration amounted to 7.4 to 1.3 depending on the concentration of NA administered (3 x 10(-9) to 10(-6) M). These concentration differences were abolished by inhibitors of uptake1 [desipramine (DMI)] and uptake2 (O-methyl-isoprenaline (OMI)). Neuronal uptake1 was characterized by a Km of 0.22 mumol/l and a Vmax of 370 pmol x min-1 x gWWT-1, extraneuronal uptake2 by a KUPTAKE of = 0.313 min-1. The apparent permeability surface area (P x S)-product calculated from uptake rate and transcapillary concentration difference was significantly decreased by administrating 100 mumol/l (NA) in presence of DMI. A presumed endothelial uptake mechanism contributing to catecholamine translocation was investigated in endothelial cells in culture. These cells showed a specific noradrenaline uptake with a K(m) of 4.35 mumol/l and a Vmax of about 75 pmol x min-1 x gWWT-1. Any inhibition by inhibitors of both of the two noradrenaline uptakes was lacking. The uptake rate of this mechanism is insufficient to contribute to the diffusive conductivity of the capillary wall (P x S-product). We conclude from our investigations on interstitial concentrations of catecholamines and transcapillary concentration differences, that the capillary wall, owing to its metabolic and diffusional characteristics, influences the exchange of catecholamines to a substantial and physiologically relevant extent.

Tissue endothelin-1 levels in perfused rat heart following stimulation with agonists and in ischaemia and reperfusion

The purpose of this study was to compare coronary and interstitial endothelin-1 (ET 1) levels in perfused rat hearts under several experimental conditions. because the cardiac tissue concentration of ET-1 is not clear. Hearts were perfused in an upside-down position with a colloid-free buffer at a constant flow rate of 9 ml/min/g heart wet weight, and immunoreactive ET 1 was determined in timed collections of coronary effluent and interstitial transudate produced by the ventricles and appearing on their surface. Basal ET-1 release into effluent was 0.26±0.007pg/min/g. and 0.005±0.0012 pg/min/g in transudate. Basal ET-1 concentration was 0.11 ±0.005 pg/ml (transudate) and 0.03±0.002 pg/ml (effluent), indicating four-fold higher transudate than effluent levels ( P<0.05). Following perfusion of hearts with collagenase to remove endothelial cells, ET-1 release into effluent was reduced to one-third and completely abolished in transudate, indicating that the peptide originated from the vascular endothelium. Perfusion of hearts with angiotensin II (0.1 μmol/l) or thrombin (5 U/ml) increased coronary perfusion pressure and ET-1 secretion, but little affected the transudate/effluent ET-1 concentration ratio (5.5 and 3.2, respectively). When coronary flow was reduced to ischaemic level (1 ml/min/g over several hours), ET-1 secretion rates into effluent were decreased by 55–65%, but increased three- to four-fold on reperfusion at normal flow ( P<0.05). The ET-1 concentrations in both fluids were still always below 1 pg/ml. No change in coronary perfusion pressure compared to time-matched normoxic controls was observed. In the presence of the ET-1 converting enzyme inhibitor, phosphoramidon (1.7 μmol/I). ischaemia-induced increases of ET-1 secretion were attenuated, and this was accompanied by a time-dependent rise in coronary perfusion pressure up to 60% ( P<0.05). These are the first measurements of endogenous cardiac tissue ET-1 levels: they do not support a vasoconstrictor (pro-ischaemic) action of endogenous ET 1 in rat hearts following ischaemia/ reperfusion, but rather point to a possible vasodilator role of the peptide under these conditions.

Maturational differences in coronary flow and interstitial transudate adenosine during alteration of perfusate oxygenation in isolated rabbit hearts.

This study was designed to investigate effects of perfusate oxygenation and maturation on coronary flow (CF), interstitial transudate adenosine (ITA) and coronary effluent adenosine (CEA) in isolated rabbit hearts. Hearts were paced at a fixed rate and were perfused under constant pressure at two different levels of perfusate oxygenation: baseline (B) (pO2 = 408 +/- 7 mmHg, O2 content = 1.28 +/- 0.03 ml O2/dl) and a lower level (L) (pO2 = 189 +/- 4 mmHg, O2 content = 0.59 +/- 0.02 ml O2/dl). CF was higher in immature (I, age 5 weeks) compared with mature (M, age 12 weeks) hearts at both levels of perfusate oxygenation (9.7 +/- 0.4 vs. 7.7 +/- 0.3 and 12.9 +/- 0.4 vs. 9.2 +/- 0.3 ml/min/g). I hearts had correspondingly higher values for myocardial oxygen consumption (MVO2) (53 +/- 3 vs. 49 +/- 2 and 39 +/- 3 vs. 35 +/- 2 microL O2/min/g), but similar values for venous oxygen tensions (nuO2) (240 +/- 9 vs. 219 +/- 10 and 74 +/- 4 vs. 62 +/- 3 mmHg), compared with M hearts at B and L. Although interstitial transudate adenosine (ITA) concentration was similar in I and M hearts at B (409 +/- 75 vs. 254 +/- 39 nM), it was lower in I than M hearts at L (2500 +/- 770 vs. 4210 +/- 1000 nM).(ABSTRACT TRUNCATED AT 250 WORDS)

Enzyme release into the interstitial space of the isolated rat heart induced by changes in contractile performance

The aim was to investigate changes in interstitial concentration and release of creatine kinase in isolated perfused rat hearts after an experimental inotropic stimulation or after recovery from a negative inotropic intervention (low Ca2+ or high K+ buffer). Interstitial transudate emerging at the surface of the heart and venous effluent were analysed for creatine kinase. The interstitial concentration of creatine kinase was always higher, by a factor of 25 to 100 (range from 10 to 580 mU.ml-1), than the concentration in the venous effluent (close to or below the limit of detection: 0.4 mU.ml-1). Continuous stimulation with a submaximal effective concentration (8 nM) of isoprenaline for 30 min resulted in an initial transient increase in the interstitial release of creatine kinase to about 160% of the control (p < 0.05). Similarly, in a second series, three repeated (5 min) periods of inotropic stimulation also caused a significant and transient increase in the interstitial release of creatine kinase to a maximum of 180%. Change to a buffer containing 2.0 mM Ca2+ after a 60 min period of low Ca2+ perfusion (0.25 mM) led to restoration of contractile function and an immediate and transient increase in the interstitial release of creatine kinase to about 900%. After a period of cardiac arrest (16 mM K+ for 60 min), perfusion with 4 mM K+ also immediately restored cardiac function, and led to an increase in creatine kinase release of 2500%. In additional experiments dilatation of the left ventricle by inflating an intraventricular balloon during cardioplegic perfusion induced a significant fivefold increase in the interstitial release of creatine kinase, which was further enhanced 3.5-fold during the subsequent recovery period. The coincidence of an increase in or a restoration of cardiac contractile function and an increase in the interstitial enzyme release suggests that myocardial enzyme release may occur in response to physiological stimuli during different episodes of metabolic or mechanical stress, as well as under pathophysiological conditions.

Interstitial lactate and glucose concentrations of the isolated perfused rat heart before, during and after anoxia.

In isolated rat hearts perfused according to the Langendorff technique lactate and glucose concentrations were determined in the interstitial transudate and the venous effluent before, during and after periods of 15 or 30 min anoxia. The interstitial transudate emerged at the surface of the heart as a result of albumin-free perfusion. During normoxic perfusion the interstitial lactate concentration was 0.144 +/- 0.025 mmol/l (n = 6); the venous lactate concentration was 0.033 +/- 0.005 mmol/l. From the interstitial and the mean vascular concentration, together with the lactate release and the glucose uptake, the apparent permeability surface area products (P.S product) were calculated using Fick's law. The apparent P.S products for lactate and glucose were 4.6 and 3.9 ml/(min x g), respectively. During anoxia we measured a four- to sixfold increase of the interstitial lactate concentration. At the end of the anoxic periods the apparent P.S product was two- to threefold higher than during normoxia; the apparent glucose P.S product increased about fourfold. After 15 min anoxia the increases of permeability were completely reversed in the reoxygenation period. However, after a period of 30 min anoxia the apparent P.S products for lactate and glucose remained raised, which means that there was a prolonged or even irreversible increase of capillary permeability. Besides the marked transcapillary concentration difference for lactate, these data show prolonged functional alteration of the capillary wall after 30 min anoxia.

Fatty Acid Transfer Across the Myocardial Capillary Wall

Interstitial transudate of isolated isometrically working perfused rat hearts was analyzed to investigate fatty acid (FA) release and transfer across the capillary wall. Unsaturated FA were released under certain conditions. Lowering medium FA (16:0) with constant or varying FA/A-ratio (A: albumin) decreased interstitial FA-concentration down to 17% of arterial FA-concentration. This was accompanied by reduced uptake-rate. Transcapillary diffusion-resistance, unless markedly altering, cannot be responsible for this observation. Calculated diffusion-rates of FA and FA * protein-complexes in the endothelial cytoplasm and interstitium indicate that FA-transfer takes place almost exclusively bound to carrier-proteins. The apparent permeability surface-area-values of FA for transendothelial FA-transfer are three orders of magnitude higher than those for sucrose; diffusion-coefficients for FA across the endothelium are close to those in water, excluding substantial diffusion-barrier of the capillary wall. Dissociation-rate-constants calculated from experimental data are in the range of those reported for FA *albumin-complexes in vitro. Thus the observed transcapillary FA-gradients are apparently due to the limited dissociation-rate of the FA *albumin-complex in the intravascular space.

Myocardial adenosine, flow, and metabolism during adenosine antagonism and adrenergic stimulation

Relationships between interstitial transudate adenosine and coronary flow and between global adenosine formation and cytosolic metabolism were examined in constant-pressure perfused guinea pig hearts during norepinephrine (NE) stimulation and adenosine antagonism with 10 microM 8-phenyltheophylline. Basal coronary flow was 5.7 ml.min-1 x g-1, and transudate and venous adenosine levels were approximately 0.26 and 0.06 microM, respectively. During 10 min of NE stimulation (15 nM), coronary flow and adenosine levels increased, the phosphocreatine-to-inorganic phosphate ratio ([PCr]/[Pi]) declined, and ATP and pH remained stable. Despite phasic release of adenosine, coronary flow correlated dose dependently with transudate adenosine, and adenosine release was inversely related to [PCr]/[Pi] under all conditions. 8-Phenyltheophylline infusion attenuated functional hyperemia by approximately 40%, enhanced the fall in [PCr]/[Pi], and potentiated elevations in transudate and venous adenosine. Similar results and correlations were obtained in hearts perfused at a constant-flow of 5.7 ml.min-1 x g-1, although stimulated adenosine levels and metabolic changes were greater and contractile responses smaller. These data indicate that: 1) endogenous adenosine plays a primary role in functional hyperemia in perfused guinea pig heart; 2) global adenosine formation appears related to phosphorylation status; and 3) adenosine receptor antagonism enhances metabolic disturbances during adrenergic stimulation and markedly potentiates adenosine release, indicating that the functional effects of antagonists may significantly underestimate the dilatory role of endogenous adenosine.

Interstitial Transudate Purines in Normoxic and Hypoxic Immature and Mature Rabbit Hearts

Interstitial transudate and coronary venous concentrations of adenosine, inosine, and hypoxanthine were determined in isolated isovolumic immature and mature rabbit hearts during normoxia and hypoxia. During normoxia, interstitial transudate adenosine was lower in immature hearts compared with mature hearts. Interstitial transudate concentrations of adenosine, inosine, and hypoxanthine were 130 +/- 16 nM, 699 +/- 88 nM, and 392 +/- 80 nM, respectively, in immature rabbit hearts and 228 +/- 35 nM, 1154 +/- 126 nM, and 287 +/- 30 nM, respectively, in mature rabbit hearts. Interstitial transudate adenosine was significantly lower in the immature hearts. Coronary venous purine concentrations were 6- and 8-fold lower than their respective interstitial transudate concentrations during normoxia in both age groups. Hypoxia significantly increased interstitial transudate purines in both age groups. Interstitial transudate adenosine, inosine, and hypoxanthine increased to 1180 +/- 231 nM, 4049 +/- 500 nM, and 1099 +/- 98 nM, respectively, in immature hearts and to 1225 +/- 300 nM, 5220 +/- 1217 nM, and 876 +/- 147 nM, respectively, in mature hearts. The age-related difference in transudate adenosine levels present during normoxia was not detected during hypoxia. Venous purine levels increased during hypoxia and the gradient from interstitial transudate fluid to venous effluent was abolished for adenosine in both groups. In immature hearts, hypoxia led to higher venous effluent adenosine levels than in the mature hearts. Coronary resistance correlated with interstitial transudate adenosine in both groups, although immature hearts displayed lower resistances at all adenosine levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Interstitial transudate concentration of adenosine and inosine in rat and guinea pig hearts.

Interstitial transudate (IT) was sampled from the surface of isolated constant pressure-perfused guinea pig and rat hearts. With endogenous adenosine (AR) formation, IT concentrations (Crr) of AR and inosine (IR) were 4- to 6.5-fold higher than those in the venous effluent. The AR-to-IR ratio varied between 0.5 and 0.1. During normoxic perfusion, CIT-AR reached a basal level of 0.18 microM. During maintained hypoxia, CIT-AR was elevated only initially up to 0.63 microM. Subsequently, it decreased to basal values, whereas coronary flow remained elevated. With repetitive hypoxia, CIT-AR decreased to basal values, with little alteration in the coronary flow response. Addition of 1,000 U/l adenosine deaminase reduced CIT-AR below 0.2 microM, with no change in coronary flow response to hypoxia. High concentrations of coformycin (Streptomyces antibioticus), an adenosine deaminase inhibitor (greater than 1 microM), were necessary to increase the AR-to-IR ratio to unity, indicating an intracellular site of action. During administration of 0.1 microM dipyridamole, no close correlation between CIT-AR and coronary flow was found. Administration of 1 microM AR did not induce a detectable change in Crr-AR despite a distinct coronary flow response. We conclude that at least in our heart preparation, interstitial adenosine seems to play no primary role in coronary flow regulation.

Intra- and extracellular markers in interstitial transudate of perfused rat hearts

The interstitial transudate was investigated in isolated perfused rat hearts. Capillary permeability and the kinetics of interstitial uptake and release were characterized using four different marker molecules (mol wt 522 to 2 X 10(6)). The half-time (t1/2) values (less than 30 to 170 s) and the interstitial concentration after 30 min (100-44% of arterial concentration) reflected the order and inverse order of their molecular weights, respectively. Creatine kinase (CK) and glutathione (GSH) were measured during control state, hypoxia, and anoxia, followed by reoxygenation. Interstitial concentrations of CK and GSH were higher by a factor of 100 and 8, respectively, compared with the venous effluent. During hypoxia (PO2 = 110 mmHg, i.e., O2 supply = 30% of demand) and reoxygenation there was a significant increase only in the interstitial (not venous) release of CK and GSH, which was further increased during anoxia. Ischemia (75 min) and reperfusion cause no interstitial release of lysosomal (acid phosphatase) and mitochondrial (glutamate dehydrogenase) enzymes despite a massive loss of cytosolic enzymes. Examination of the interstitial transudate allows characterization of capillary transfer and provides a very sensitive measure of sarcolemmal release phenomena.

Comparative pathophysiology of pulmonary hypertension: development of a model.

The lesser circulation is affected by a considerable variety of diseases that involve the heart or lungs. Blood flow in the pulmonary vessels may be compromised by destruction of lung parenchyma and excessive positive alveolar pressure as in chronic obstructive lung disease, by frank obstruction of vessels, large or small, as in pulmonary thrombo-embolic disease, by obstruction to the outflow of blood as in diseases of the pulmonary veins, mitral valve or left ventricle, or by changes in the arteries due to chronic overloading as in several congenital heart conditions with left-to-right shunts. The response of the pulmonary circulation is conditioned by the necessity of maintaining sufficient perfusion of the lungs for effective gas exchange to take place, without allowing hydrostatic pressures in the capillaries to exceed plasma oncotic pressure so much as to result in frank edema. Ignoring for the moment the role of the lymphatics in draining away interstitial transudate to dispel such edema, it seems reasonable that one of the responses of the pulmonary arteries will be to protect the capillary bed by developing increased resistance to perfusion.