Biomedical subjects

A Deussen

Publications and source records attributed to A Deussen.

At least 37 records · Page 2Linked to original sources

PubMed1997

Determinants of the S-adenosylhomocysteine (SAH) technique for the local assessment of cardiac free cytosolic adenosine.

The S-adenosylhomocysteine (SAH) technique allows the estimation of the free cytosolic adenosine concentration using the kinetic properties of the enzyme SAH-hydrolase (adenosine+homocysteine reversible SAH+H2O). Besides the cytosolic adenosine concentration, the local SAH signal may also depend on the local homocysteine availability, the continuous production of SAH from S-adenosylmethionine (SAM-->SAH+CH3) and the activity of the enzyme SAH-hydrolase. These variables were studied with high spatial resolution (sample dry mass 25 mg) in left ventricular myocardium from 26 anesthetized open-chest dogs in which heart rate averaged 86 +/- 14 beats/min and mean aortic pressure 96 +/- 17 mmHg. Homocysteine infusion (48 mg/kg i.v.) increased the normal plasma homocysteine concentration from 5.0 +/- 0.8 to 586 +/- 40 microM after 30 min when the average tissue concentration was 94% of the plasma concentration and similar in low and high flow areas (flow range 0.04 to 1.91 ml/min/g). Local SAH content was 1.18 +/- 0.48 nmol/g under control conditions and increased to 4.33 +/- 0.59 nmol/g within 60 min following competitive blockage of the SAH-hydrolase by adenosine dialdehyde (10 mumol/kg i.v.). This increase of the SAH content was slightly more in high than in low-flow areas (P < 0.01). Regional SAH-hydrolase activity (9.0 +/- 0.5 nmol/min/g) was comparable in high and low flow areas. All three variables exhibited an observed variability which was larger than the methodical variability suggesting significant spatial heterogeneity in the myocardium. A regrouping analysis indicated that between four and five samples taken from distant sites should be averaged to obtain a robust estimate of the above metabolic parameters. Reconciling the measurements with a mathematical model of cardiac adenosine metabolism and fitting of the measured SAH tissue levels gave an estimate of 72 pmol/min/g for the mean transmethylation rate. Estimates of the cytosolic adenosine concentration of cardiomyocytes and endothelial cells under control physiological conditions were 24 and 7 microM, respectively. Thus, the present measurements provide a basis for the quantitative assessment of the local cytosolic adenosine concentration in relation to blood flow.

Adenosine↗

PubMed1997

Local myocardial glucose uptake is proportional to, but not dependent on blood flow.

In nonischaemic myocardium local deoxyglucose deposition is proportional to local blood flow (y = 0.77x + 0.25 for normalized deoxyglucose deposition and blood flow). The cause for this relationship was evaluated using a mathematical model of (deoxy)glucose metabolism to elucidate whether differences in local deoxyglucose deposition are dependent on local blood flow or whether they reflect differences of the local metabolic rate. The axially distributed model consists of two blood-tissue exchange regions arranged concentrically representing the capillary and the extracapillary regions, the latter being a composite of the endothelial, interstitial and parenchymal cell regions. Exchange between the two model regions is described by a permeability-surface area product, consumption in the extracapillary region is modelled by an intraregional clearance term. Twenty blood-tissue exchange units are arranged as parallel pathways to account for the effects of flow heterogeneity and in series with a single, nonexchanging, large vessel segment (0.1 ml/g). Regional volumes (capillary 0.07 ml/g, extracapillary 0.60 ml/g) and the permeability-surface area product (0.2 ml min-1 g-1) are taken from published sources, the local myocardial blood flow is that determined experimentally (between 0.1 and 2.5 ml min-1 g-1). The only free parameter in the model is the extracapillary clearance term (0.0275-0.102 ml min-1 g-1) which was used to fit the measured tissue radioactivity concentration taking the measured arterial concentration of 3H-deoxyglucose into account. The results obtained indicate that variations in local myocardial blood flow, and hence differences in deoxyglucose supply, can not explain quantitatively the differences in local deoxyglucose deposition. However, differences of the metabolic rate (0.326 versus 0.120 micro;mol min-1 g-1) assumed to occur in parallel to the flow differences (1.26 versus 0.42 ml min-1 g-1) can well explain the different mean deoxyglucose deposition in high and low flow areas. This result supports the view that blood flow heterogeneity in the heart is paralleled by a spatial heterogeneity of glucose metabolism.

Animals↗

PubMed1997

Minimal effects of nitric oxide on spatial blood flow heterogeneity of the dog heart.

Eleven Beagle dogs were studied to elucidate the possible role of L-arginine-derived nitric oxide on local blood flow distribution in left and right ventricular myocardium. Local blood flow was determined in 256 samples from the left and 64 samples from the right ventricle per heart using the tracer microsphere technique (mean sample mass 319 +/- 131 mg). Nitric oxide production was effectively inhibited by intravenous infusion of 20 mg/kg nitro-L-arginine methylester (L-NAME) as evidenced by a shift of the dose/response curve for the effect of intracoronary administration of bradykinin (0.004-4.0 nmol/min) on coronary blood flow. L-NAME enhanced left and right ventricular systolic pressures from 132 +/- 18 to 155 +/- 15 mm Hg and from 26 +/- 3 to 29 +/- 3 mm Hg respectively (both P = 0.043). Mean left ventricular blood flow was 1.14 +/- 0.38 before and 0.99 +/- 0.28 ml min-1 g-1 after L-NAME (P = 0.068), while right ventricular blood flow fell from 0.72 +/- 0.28 to 0.53 +/- 0.20 ml min-1 g-1 (P = 0.043). Coronary conductance of left and right ventricular myocardium fell by 31 and 43% respectively (both P = 0.043). The coefficient of variation of left ventricular blood flow was 0.26 +/- 0.07 before and 0.29 +/- 0.07 after L-NAME (P = 0.068), that of right ventricular blood flow was 0.27 before and after L-NAME. Skewness (0.51) and kurtosis (4.23) of left ventricular blood flow distribution were unchanged after L-NAME, while in the right ventricle skewness decreased from 0.54 to 0.09 (P = 0.043) and kurtosis (3.68) tended to decrease after L-NAME (P = 0.080). The fractal dimension (D = 1.20-1.27) and the corresponding nearest-neighbor correlation coefficient (rn = 0.37-0.53) of left and right ventricular myocardium remained unchanged after infusion of L-NAME. From these results it is concluded that firstly, local nitric oxide release does not explain the higher perfusion of physiological high flow samples and secondly, that spatial myocardial blood flow coordination is not dependent on nitric oxide.

Animals↗

PubMed1996

Characterisation of left ventricular relaxation in the isolated guinea pig heart.

The time constant of left ventricular pressure fall, tau, has frequently been used as a measure of myocardial relaxation in the blood-perfused, ejecting heart. The aim of the present study was to characterise tau in relation to beta-adrenergic activation, coronary perfusion pressure and flow as well as cardiac oxygen supply and demand in the isolated, isovolumically beating heart. Therefore, tau was analysed from digitised left ventricular pressure data in a total of 23 guinea pig hearts perfused with saline at constant pressure (60 cmH2O). The coronary venous adenosine concentration ([ADO]) served as an index of myocardial oxygenation. Isoprenaline (0.4-3.2 nmol l-1) decreased and propranolol (3-9 mumol l-1) increased tau dose-dependently (linear regression tau vs lg([isoprenaline]), r = 0.74; tau vs. lg([propranolol]), r = 0.66, both P < 0.05). During graded reductions in cardiac oxygen supply from 96.1 +/- 12.6 (SEM) to 44.4 +/- 4.4 microliters min-1 g-1, tau was prolonged from 61.5 +/- 12.7 to 109.9 +/- 22.6 ms while left ventricular developed pressure (LVDP) decreased from 90.7 +/- 7.2 to 40.7 +/- 5.1 mmHg. In parallel, [ADO] increased from 23.7 +/- 9.1 to 58.0 +/- 19.1 pmol ml-1 (P < 0.05). Increasing oxygen supply to 165.4 +/- 32.4 microliters min-1 g-1 augmented LVDP to 102.7 +/- 7.3 mmHg but did not change tau or [ADO]. There was a dual response of tau to changes in cardiac oxygen supply or demand. As long as oxygen supply and demand matched, tau remained constant. However, when the oxygen supply was less than 100 microliters min-1 g-1, left ventricular relaxation was prolonged in parallel to the reduction in oxygen supply. In addition, a close relationship was observed between [ADO] as an indicator of myocardial oxygenation and tau (Spearman correlation, r = 0.99, P < 0.005). We conclude that the time constant of left ventricular pressure fall, tau, sensitively reflects myocardial relaxation in the isolated, isovolumically beating guinea pig heart. Moreover, in this model left ventricular relaxation is not influenced by alterations in coronary perfusion pressure or flow as long as cardiac oxygen demand is matched by an adequate supply. Rather relaxation is strictly coupled to myocardial oxygenation as reflected by coronary venous adenosine concentrations.

Animals↗

PubMed1996

Spatial heterogeneity of blood flow in the dog heart. I. Glucose uptake, free adenosine and oxidative/glycolytic enzyme activity.

The spatial heterogeneity of myocardial perfusion and metabolism was studied in 11 anaesthetized dogs under resting conditions. In each heart local myocardial blood flow was assessed using the tracer microsphere technique in 256 samples (mean mass: 83.1 mg) taken from the left anterior ventricular wall. In the same samples, the following biochemical parameters were determined: accumulation of [3H]-deoxyglucose (a measure of glucose uptake), free cytosolic adenosine (S-adenosylhomocysteine accumulation technique, a measure of tissue oxygenation and a possible mediator of blood flow regulation), and the specific activities of oxidative (citrate synthase, cytochrome-c-oxidase) and glycolytic (hexokinase, phosphoglycerate kinase) enzymes. Capillary density and mitochondrial and myofibril volume densities were determined by morphometry. Myocardial perfusion in each sample (average 0.77 ml min-1 g-1) varied between 0.1 and 2.5 times the mean (coefficient of variation 0.30+/-0.02). [3H]-deoxyglucose was deposited locally in proportion to perfusion. Samples showing low flow (<0.2 ml min-1 g-1) did not exhibit increased levels of cytosolic adenosine. The specific activities of the oxidative and glycolytic enzymes, however, were uniformly distributed between low and high flow areas. Furthermore, capillary density and mitochondrial and myofibril densities were similar in high and low flow regions. The results show firstly that local glucose metabolism in the heart occurs in proportion to local blood flow, suggesting that high flow regions have a higher than average metabolic rate. Secondly, regions of low flow are not compromized by critical oxygenation and most likely have a lower than average oxygen demand and finally, the homogeneous distribution of oxidative and glycolytic enzymes, as well as the homogeneous myocardial ultrastructure, suggest that areas with high and low blood flow under resting conditions may increase their metabolic rate to similar levels when required.

Adenosine↗

PubMed1996

Spatial heterogeneity of blood flow in the dog heart. II. Temporal stability in response to adrenergic stimulation.

The effects of adrenergic stimulation on local myocardial blood flow in the left ventricle were studied in 13 anaesthetized Beagle dogs using the tracer microsphere technique. Adrenergic stimulation was induced by intravenous infusion of orciprenaline (1-2 microg kg-1 min-1) over 15 min or by electrical stimulation of the left ansa subclavia (10 Hz, 1 ms, 4-8 V) over 5 min. Local myocardial blood flow was analysed in 256 samples with an average (+/-SD) mass of 318+/-49 mg from the left ventricular myocardium using a standardized dissection procedure. Orciprenaline increased the average myocardial blood flow from 0.85+/-0.18 to 1.73+/-0.27 ml min-1 g-1, while oxygen consumption and the pressure-rate product increased by 129 and 119% respectively. The coefficients of variation of local myocardial blood flow, a measure of spatial blood flow heterogeneity, were 0.21 and 0.18 under control and orciprenaline respectively. Except for a slight transmural gradient (endomyocardium/epimyocardium flow ratio 1.19) myocardial blood flow did not exhibit significant spatial gradients. Stimulation with orciprenaline increased the average blood flow in all regions of the left ventricle by comparable extents. However, local blood flow during orciprenaline was significantly lower in samples from regions which had a lower blood flow under resting control conditions. A significant positive relationship was obtained between local myocardial blood flow under resting conditions and orciprenaline (r=0.45+/-0.18). Moreover, after recovery from orciprenaline stimulation (i.e. 40-112 min after the end of orciprenaline infusion) local myocardial blood flow exhibited a high degree of correlation with local flow before orciprenaline (r=0.71+/-0.08). Comparable results were obtained with electrical stimulation of the left ansa subclavia. For the comparison stimulation vs. control, the correlation coefficient of local blood flow was 0.52+/-0.04 and for recovery vs. control 0.77+/-0.06. From these results it is concluded firstly that local myocardial blood flow under resting conditions is an important determinant of local flow during adrenergic stimulation. Secondly, the anatomical region does not have any predictive value for the blood flow change during adrenergic stimulation and finally, the close relationship between local blood flow before and after cardiac stimulation indicates that the spatial blood flow heterogeneity is temporally stable over hours.

Adrenergic Agonists↗

PubMed1996

Modeling [15O]oxygen tracer data for estimating oxygen consumption.

The most direct measure of oxidative tissue metabolism is the conversion rate of oxygen to water via mitochondrial respiration. To calculate oxygen consumption from the analysis of tissue residue curves or outflow dilution curves after injection of labeled oxygen one needs realistic mathematical models that account for convection, diffusion, and transformation in the tissue. A linear, three-region, axially distributed model accounts for intravascular convection, penetration of capillary and parenchymal cell barriers (with the use of appropriate binding spaces to account for oxygen binding to hemoglobin and myoglobin), the metabolism to [15O]water in parenchymal cells, and [15O]water transport into the venous effluent. Model solutions fit residue and outflow dilution data obtained in an isolated, red blood cell-perfused rabbit heart preparation and give estimates of the rate of oxygen consumption similar to those obtained experimentally from the flow times the arteriovenous differences in oxygen contents. The proposed application is for the assessment of regional oxidative metabolism in vivo from tissue 15O-residue curves obtained by positron emission tomography.

Animals↗

PubMed1995

Observation of reorientationally hindered water in biological tissue using triple quantum filtered 17O-NMR.

Water dynamics in aqueous biopolymer solutions often display a two-phase character, resembling water-water and water-protein interactions. Rotationally hindered water molecules in crowded protein environments display triple exponential magnetic relaxation out of the extreme narrowing limit. Because water-protein interactions retard the water dynamics, H2O(17) magnetization passes through a NMR multiple quantum coherence filter, allowing the visualization of reorientationally hindered water without the disturbing resonance of the bulk. In vitro experiments performed on selected biological materials (lens, vitreous body and serum albumin solutions) clearly demonstrate the potential of this technique.

Animals↗

PubMed1995

Role of nitric oxide in the regulation of coronary vascular tone in hearts from hypertensive rats. Maintenance of nitric oxide-forming capacity and increased basal production of nitric oxide.

In arterial hypertension, coronary flow reserve, expressed by the difference between autoregulated and maximal coronary flow, is frequently impaired. Previous experimental and clinical investigations using acetylcholine as a stimulus for the production of endothelium-derived relaxing factor suggested that an impaired endothelium-dependent vasodilation, presumably caused by a decreased formation of nitric oxide (NO), may account for this microvascular dysfunction. However, so far no study has been performed that quantifies the formation of NO within the coronary circulation of hypertensive hearts to assess its role in setting coronary vascular tone in the hypertensive heart. We therefore quantified NO formation within the coronary circulation of constant flow-perfused, isolated hearts from spontaneously hypertensive rats (SHR, 16th to 26th week), as a model for hypertensive heart disease, and from the normotensive control strain (Wistar-Kyoto, WKY) using the oxyhemoglobin technique. Coronary perfusion pressure and vascular resistance were almost 30% higher in SHR compared with WKY hearts. Intracoronarily applied NO decreased coronary vascular resistance by maximally 45% of resting values in a concentration-dependent manner in both groups. The bradykinin-induced decrease in coronary vascular resistance and the parallel increase in NO release were comparable in SHR and WKY hearts and fell within the vasodilator range of exogenously applied NO. Moreover, basal release of NO normalized to heart wet weight was 50% higher in SHR compared with WKY hearts. Rates of basal NO release were correlated inversely with changes in coronary perfusion pressure and vascular resistance in both groups (r = -.85 and -.84, respectively, P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Understanding the controversy over the identity of EDRF.

Thirteen years after its discovery, there is still controversy over the chemical identity of endothelium-derived relaxing factor (EDRF). Although pharmacological and chemical evidence indicates that EDRF is nitric oxide, other candidates, including S-nitrosocysteine, dinitrosyl-iron-cysteine complex, nitroxyl and hydroxylamine, have been proposed to account for the vasorelaxant properties of EDRF. Such diverse compounds should differ in their stability and in reactivity with oxyhaemoglobin and with redox-active nucleophiles such as thiols. Here we use a bioassay to compare the pharmacodynamic profiles of these and other compounds with those of nitric oxide and EDRF. We find that some S-nitrosothiols, dinitrosyl-iron-cysteine complex, sodium nitroxyl and hydroxylamine can be eliminated as candidates as they are more stable than EDRF and less susceptible to inhibition by oxyhaemoglobin. Co-infusion of cysteine revealed major differences between the remaining candidates because it reduced the effect of authentic nitric oxide and EDRF on the bioassay tissues but enhanced the survival of S-nitrosocysteine and S-nitrosocysteamine. Our results further support the evidence that EDRF, the pharmacological entity described by Furchgott and Zawadzki, is nitric oxide.

Animals↗

PubMed1994

[Effect of arginine-dependent nitric oxide synthesis on regional perfusion of the eye in the anesthesized dog].

The changes of the retinal and uveal perfusion after inhibition of the arginine-dependent nitric oxide (NO) synthesis by systemic administration of NG-nitro-L-arginine methyl ester (L-NAME; 20 mg/kg bw i.v.) were studied in four anesthetized dogs using the tracer microsphere technique. The regional perfusion rates (ml.min-1.g-1) under steady-state control conditions were: retina 0.13 +/- 0.05, choroid 8.26 +/- 1.85, iris 0.24 +/- 0.05 and ciliary body 1.11 +/- 0.26. Infusion of L-NAME over 10 min reduced the perfusion of the retina on the average by 23% (P > 0.05). The perfusion of choroid, iris and ciliary body fell by 54 +/- 8%, 58 +/- 7% and 53 +/- 8%, respectively (P < or = 0.05 for all). In five additional experiments the local activity of NO-producing enzymes (NO synthases) was determined by measuring the production rate of citrullin in tissue extracts of the different eye regions. Total NO synthase activities (pmol citrullin.min-1.g-1) were: retina 31.0 +/- 5.5, choroid 3.1 +/- 2.8, iris 7.1 +/- 2.1 and ciliary body 1.3 +/- 1.3. Differences of the total NO synthase activities of retina, iris and cilary body were statistically significant (P < or = 0.02). The results show that the uvea perfusion is largely influenced by the steady-state production of NO. The homogeneous flow reduction in the uvea after inhibition of NO synthase is contrasted by the heterogeneous NO synthase activities of the different uvea regions.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Oxidoreductases↗

PubMed1993

L-arginine-derived nitric oxide: a major determinant of uveal blood flow.

The effect of inhibition of the L-arginine-nitric oxide pathway on regional ocular blood flow was studied in anaesthetized Beagle dogs. Under control conditions (mean arterial blood pressure 105 +/- 6 mmHg) trapping of 11 and 15-microns-diameter tracer microspheres revealed the following regional blood flows (ml min-1 g-1): retina 0.065 +/- 0.016, choroid 5.72 +/- 0.32, ciliary body 0.77 +/- 0.11, iris 0.18 +/- 0.04. After i.v. infusion of nitro-L-arginine methylester (20 mg kg-1), a potent inhibitor of nitric oxide production from L-arginine, mean arterial blood pressure increased from 105 +/- 6 mmHg by 19% to a new steady-state level of 125 +/- 7 mmHg. This increase of arterial blood pressure extended over 3 hr and was reversible after i.v. infusion of L-arginine (100 mg kg-1 over 10 min). Despite the increase in blood pressure following infusion of nitro-L-arginine methylester blood flow of choroid, ciliary body, and iris were significantly decreased by 40, 40 and 48%, respectively. Retinal blood flow did not change significantly (-12%). These results suggest that the L-arginine-nitric oxide pathway is of major importance for the adjustment of uveal blood flow under resting physiological conditions.

Animals↗

PubMed1993

Formation and salvage of adenosine by macrovascular endothelial cells.

Contribution of extracellular adenine nucleotide degradation to adenosine formation and internal salvage of adenosine via adenosine kinase were quantified in macrovascular porcine endothelial cells. Microcarrier beads covered with endothelial cells were kept in a perfusion column at a flow rate of 2 ml/min. Total adenine nucleotide (AN) release was quantified with a sensitive firefly luciferin-luciferase assay after enzymatic rephosphorylation of AMP and ADP to ATP. Adenosine (ADO) was measured by radioimmunoassay or high-pressure liquid chromatography (HPLC) techniques. Basal AN and ADO release under steady-state conditions were 2.2 and 13.8 pmol.min-1 x ml column volume (CV)-1, respectively. Inhibition of adenosine deaminase with erythro-9-(2-hydroxy-3-nonyl)adenine (5 x 10(-6) M) enhanced ADO release by 3.3 pmol.min-1 x ml CV-1, and AN release remained unchanged (2.8 pmol.min-1 x ml CV-1). Inhibition of adenosine kinase by 5-iodotubercidine (10(-5) M) greatly enhanced ADO release by 97.7 pmol.min-1 x ml CV-1, while AN release was unaffected. Inhibition of ecto-5'-nucleotidase by alpha,beta-methylene-ADP (5 x 10(-5) M) enhanced AN release from 2.6 to 8.2 pmol.min-1 x ml CV-1 and reduced ADO release by an equivalent extent. Stimulation of endothelial cells with Ca ionophore A23187 dose dependently augmented AN and ADO release to 2,013.2 and 92.5 pmol.min-1 x ml CV-1, respectively. Thrombin (1 U/ml) enhanced AN release from 5.0 to 8.7 pmol.min-1 x ml CV-1, whereas several other endothelium-dependent and -independent vasodilators including acetylcholine, bradykinin, isoproterenol, and norepinephrine were proven to have no significant effect.(ABSTRACT TRUNCATED AT 250 WORDS)

5'-Nucleotidase↗

PubMed1993

A vascular transport operator.

A pulse or a sharp front in concentration of a tracer or a substrate in the blood within a vessel becomes dispersed while being transported along a vessel. Cross-stream mixing and pulsations in flow with the heartbeat cause the dispersion to be less than would occur with a parabolic velocity profile (Newtonian flow). These characteristics allow intravascular mass transport to be described well by a simple two-parameter differential operator, which is a one-dimensional representation of the rather complex real situation. The operator consists of two components in series, a pure delay and a fourth-order linear differential operator. The latter is merely two underdamped second-order operators in series, with fixed relationships between the natural frequencies and damping coefficients. The operator is useful because it provides a transport function with skewness and kurtosis suitable to intravascular transport where the mean velocity profile is blunter than in Newtonian parabolic flow. The parameters of the operator are its mean transit time, t, and its relative dispersion, RD, which is the standard deviation of the response impulse divided by t. The operator transport function describes blood transport through the human leg arterial system, where the RD values are approximately 15-20%.

Arteries↗

PubMed1992

Role of nitric oxide in local blood flow control in the anaesthetized dog.

Intravenous infusion of NG-nitro-L-arginine methyl ester (L-NAME), a potent inhibitor of nitric oxide (NO) formation from L-arginine, provokes marked rises in arterial blood pressure by increasing peripheral resistance. In order to further evaluate the contribution of basal NO-formation to control of organ blood flow, regional blood flow distribution within the myocardium, kidney and brain areas was assessed using the tracer-microsphere technique in anaesthetized dogs. After L-NAME (20 mg kg-1 i.v.) kidney perfusion was homogeneously reduced by 55% in the entire cortex and the outer medulla. Within the left ventricular myocardium regional blood flow significantly decreased only in subepicardial layers (-12%), whereas within the entire right ventricle regional blood flow was reduced by 19-24%. A close inverse relationship was found between all changes in regional myocardial blood flows observed after L-NAME and the respective control values. No significant changes in regional blood flow in different areas of the brain were detectable after L-NAME. It is concluded that the contribution of basal NO formation varies greatly between different organs and exhibits significant regional differences within the heart. It is possible that local metabolic mechanisms may compensate functionally for the inhibition of NO synthesis.

Animals↗

PubMed1992

S-adenosylhomocysteine hydrolase activity in human myocardium.

OBJECTIVE: Measurement of S-adenosylhomocysteine (SAH) accumulation in the heart reflects the concentration of free cytosolic adenosine and is thus a sensitive indicator of regional myocardial ischaemia. To evaluate the possibility of applying this method in combination with 11C-SAH positron emission tomography (PET) to patients with ischaemic heart disease the activity of SAH hydrolase in human heart muscle and its regional distribution were studied. METHODS: Myocardium from patients with dilated cardiomyopathy (n = 4), hypertrophic obstructive cardiomyopathy (HOCM, n = 6), and mitral stenosis (n = 3) was analysed. Additional studies were performed in myocardium, isolated cardiomyocytes, and endothelial cells from dog and guinea pig hearts. Enzyme activity in synthetic and hydrolytic direction including kinetic data (Vmax, KM values, pH dependency) was measured in the cytosolic fraction of myocardial tissue and cell extracts, using high performance liquid chromatography and photometric methods, respectively. RESULTS: Rates of SAH synthesis (Vmax) in the left ventricle in dilated cardiomyopathy, mitral stenosis, and HOCM were 0.8 (SEM 0.1), 1.0(0.2), and 1.7(0.1) nmol.min-1.mg-1 protein respectively. KM values for DL-homocysteine, adenosine, and SAH in HOCM were 187, 2.2, and 3.4 microM, respectively. Enzyme activity was homogeneously distributed among right and left atria, right and left ventricles, and septum. Additional studies in homogenated muscle and isolated cardiomyocytes of guinea pig and canine hearts showed activities similar to man. CONCLUSIONS: (1) SAH hydrolase activity in the human heart is quantitatively comparable to that found in other mammals but certain myocardial diseases may go along with changes in SAH hydrolase activity; (2) the kinetic properties, absolute amounts, and homogeneous distribution of the enzyme may permit the non-invasive determination of free adenosine in the human heart by PET.

Adenosylhomocysteinase↗

PubMed1992

Comprehensive model of transport and metabolism of adenosine and S-adenosylhomocysteine in the guinea pig heart.

Regulation of blood flow and mitochondrial respiration in the heart would be clarified by improved knowledge of interstitial concentrations and cellular production rates of adenosine; however, these variables cannot be measured directly. To interpret indexes that are available, a comprehensive mathematical model was developed, based on a large body of experimental data. The model describes most of the important pathways of capillary-tissue transport and cellular metabolism of adenosine in the guinea pig heart. It includes capillary flow, solute transport between tissue regions, nonlinear enzyme kinetics for adenosine kinase and adenosine deaminase, and reversible biunireactant kinetics for S-adenosylhomocysteine hydrolase in cardiomyocytes and endothelial cells, intracellular production of adenosine via AMP hydrolysis and transmethylation, and extracellular production of adenosine. A single set of parameter values for the model was obtained in the first stage of the analysis by taking certain values directly from published sources, other values were subject to specific constraints, and other values were determined by parameter optimization. The effects of flow and endothelial metabolism on the relation between interstitial and venous adenosine concentrations were determined. The relation between myocardial adenosine production rate and S-adenosylhomocysteine accumulation in the presence of excess homocysteine was estimated. In the second stage of the analysis, the model was used to investigate the mechanism of myocardial adenosine production, without changing the parameter values. Cellular adenosine production rates were estimated by fitting measurements of venous adenosine release obtained during altered energetic conditions in experiments by different investigators. The original results showed a dissociation between measurements of cytosolic AMP concentrations and venous adenosine release. It is concluded that 1) it is essential to account for the effect of flow on interstitial and venous adenosine concentrations, since decreased flow may produce effects outwardly resembling inhibition of the enzyme 5'-nucleotidase, 2) adenosine concentrations in epicardial transudate are not in equilibrium with interstitial fluid, and 3) the rate of cellular adenosine production increases monotonically with free cytosolic concentrations of AMP during a variety of alterations in energy balance of the guinea pig heart.

Adenosine↗

PubMed1992

Noninvasive assessment of regional cardiac adenosine using positron emission tomography.

One of the early metabolic changes associated with myocardial ischemia is the breakdown of adenine nucleotides resulting in the enhanced production of adenosine. In order to image regional cardiac adenosine by positron emission tomography (PET) the enzymatic conversion of adenosine into [11C]-S-adenosylhomocysteine ([11C]SAH) was used in the presence of 11C-labeled homocysteine thiolactone (adenosine + [11C] - homocysteine-->[11C] - SAH + H2O). Following production of an experimental coronary constriction in anesthetized dogs carrier added 1-[11C]-D,L-homocysteine thiolactone (5-27 mCi, 30 mg/kg) was infused over 1 min. This intervention, while hemodynamically ineffective, increased the plasma homocysteine concentration from 2.5 to 306 microM, which thereafter declined with a T1/2 of 28 min to 97 microM after 60 min. During the first minutes following infusion of [11C] homocysteine, the radioactivity concentration in the blood pool, the nonischemic and the ischemic myocardium were similar. Between 20 and 60 min, however, the regional radioactivity concentration was highest in the perfusion area of the stenosed vessel: 6.6% compared to 5.2 and 5.2% of the injected dose per 1 I tissue. The elevated radioactivity concentration was strictly confined to the perfusion area of the occluded artery. Using [35S]-L-homocysteine (20 microCi; 30 mg/kg) chromatographic separation of SAH in tissue extracts confirmed that the radioactivity accumulation was due to trapping of adenosine in the cellular SAH-pool. These experiments provide first evidence that 1-[11C]homocysteine thiolactone can be successfully used to assess regional adenosine formation in the heart with PET via measurement of [11C] SAH accumulation.

Adenosine↗