Importance of basal nitric oxide synthesis in regulation of myocardial blood flow.

STUDY OBJECTIVE: The aim was to investigate whether basal coronary vascular tone and myocardial perfusion depend upon endothelial nitric oxide (NO) synthesis. DESIGN: Myocardial blood flow and vascular resistance of the left and right ventricles were studied before and after intravenous infusions of either NG-nitro-L-arginine (L-NA), a specific inhibitor of NO synthase, or L-arginine, the precursor of NO synthesis. Radiolabelled microspheres were used to study myocardial blood flow in small tissue sections. EXPERIMENTAL MATERIAL: 14 anaesthetised male cats, weight 2.1-3.5 kg, were used. MEASUREMENTS AND MAIN RESULTS: Measurements were made before and 15 and 40 min after L-NA treatment (30 mg.kg-1 bolus followed by 1 mg.kg-1.min-1 infusion; n = 8), and before and 15 min after L-arginine treatment (30 mg.kg-1 bolus followed by 10 mg.kg-1.min-1 infusion; n = 6). L-NA significantly reduced coronary blood flow to the left and right ventricle, by 30(SEM 9) and 48(6)% respectively, after 15 min, but only to the right ventricle, by 45(8)%, after 40 min. Mean arterial pressure and myocardial vascular resistance were raised during the L-NA infusion. In contrast, L-arginine did not elicit any change in the variables studied. CONCLUSIONS: The conductance of the coronary vascular bed and the resting myocardial blood flow is regulated by L-arginine derived nitric oxide, and exogenous L-arginine availability is not a limiting factor in this NO generation.

Can the Indo-1 fluorescence approach measure brain intracellular calcium in vivo? A multiparametric study of cerebrocortical anoxia and ischemia.

Indo-1 fluorescence was used to monitor intracellular calcium levels in the cat brain in vivo, using the approach proposed by Uematsu et al. [Uematsu D., Greenberg J. H., Reivich M., Karp A. In vivo measurement of cytosolic free calcium during cerebral ischemia and reperfusion. Ann Neurol 1988; 24: 420-428]. In addition, extracellular calcium and potassium levels, NADH redox state, electrocorticogram (ECoG), DC potential and relative cerebral blood flow were monitored simultaneously. Changes in the Indo-1 fluorescence ratio F400/F506 were monitored during anoxia, reversible ischemia and irreversible ischemia. Although these perturbations resulted in the expected changes in extracellular calcium and potassium levels, NADH redox state, ECoG and other physiological parameters, they did not result in significant increases in the F400/F506 ratio. The apparent insensitivity of the in vivo Indo-1 approach is due to the difficulty in obtaining accurate fluorescence signals from Indo-1 in the brain. Two reasons for this difficulty appear to be problems in loading Indo-1 into the brain, and problems in correcting Indo-1 fluorescence signals for changes in NADH fluorescence and changes in absorption of intrinsic chromophores. Under the conditions of our in vivo cat experiments, Indo-1 fluorescence is not a viable approach for measuring changes in cerebral intracellular calcium levels.

Effect of topically administered epinephrine, norepinephrine, and acetylcholine on cerebrocortical circulation and the NAD/NADH redox state.

We investigated the effects of topically administered catecholamines and acetylcholine (ACh) on the cerebrocortical microcirculation and NAD/NADH redox state in chloralose-anesthetized cats. NADH fluorescence of the brain cortex and the volume of small intracortical vessels were measured by fluororeflectometry, and in most of the experiments the pial vessels were photographed simultaneously through a cranial window. Cerebrocortical vascular volume (CVV) and the diameter of the pial vessels were decreased, and NADH was oxidized by concentrations of epinephrine and norepinephrine as low as 3 x 10(-8) M. Pial veins constricted approximately twice as much as pial arteries. ACh dilatated pial arteries, slightly constricted pial veins, and increased CVV, but had no effect on the NAD/NADH redox state. Since pial and intracortical vessels were constricted markedly by catecholamines, and since these vascular reactions appeared at a lower concentration than is presumed to occur in the synaptic cleft, our results support the regulating role of these substances in cerebral circulation. NADH oxidation, obtained with catecholamines, was interpreted to be due to enhanced tissue respiration. The finding that ACh dilatated pial arteries and increased CVV, but failed to influence the NAD/NADH redox state, might indicate that the brain cortices of normal animals are bioenergetically nonhypoxic. If cortical microregions where the oxygen tension is close to zero were biochemically hypoxic, NADH oxidation should have occurred during ACh administration.

Effect of acute arterial hypo- and hypertension on cerebrocortical NAD/NADH redox state and vascular volume.

The effects of stepwise arterial hypotension (MABP: 80, 60, 40 mm Hg) and moderate arterial hypo- and hypertension (MABP: 80, 150-160 mm Hg) on cerebrocortical vascular volume and NAD/NADH redox state were studied in anaesthetized cats. The vascular volume and NADH fluorescence measurements were performed on closed skull preparations using a microscope fluororeflectometer. To determine the possible role of adrenergic alpha-receptors in the autoregulatory adjustment of cerebrocortical vascular volume, some of the animals were pretreated with intra-arterially infused phenoxybenzamine (1 mg/kg). It was found that longlasting stepwise arterial hypotension leads to a gradual increase in cerebrocortical vascular volume and NADH fluorescence. Though the cerebrocortical arteries dilatated considerably at 80 mm Hg, sustained for 30 min, the NAD/NADH redox state failed to be reoxidized but was shifted to a more reduced state. This finding suggests that some factor other than tissue hypoxia is responsible for the dilatation of cerebrocortical vessels during moderate arterial hypotension. When the arterial blood pressure was restored following stepwise arterial hypotension, the cerebrocortical vascular volume did not decrease and the NAD/NADH redox state remained reduced, showing that the autoregulatory capability of the vessels was lost and the tissue metabolism was irreversibly altered. During a 5-min duration of moderate arterial hypo- and hypertension, biphasic changes were obtained in cerebrocortical vascular volume while the NAD/NADH redox state was shifted to a more reduced and oxidized state. Since the dilatation and the constriction of the cerebrocortical vessels during arterial hypo- and hypertension lagged by 40-80 s behind the redox state alterations, it is suggested that the myogenic mechanism has a minor role in CBF autoregulation. Phenoxybenzamine (PBZ) dilatated the cerebrocortical vessels, indicating the existence of an active alpha-receptor-mediated vasoconstrictory tone. Since the extent of autoregulatory vascular volume changes was not affected by PBZ pretreatment, the involvement of adrenergic alpha-receptors in the autoregulation of CBF can be excluded, at least for cats.

Effect of topical adenosine deaminase treatment on the functional hyperemic and hypoxic responses of cerebrocortical microcirculation.

The purpose of this study was to investigate the possible importance of adenosine in cerebrocortical vasodilatation accompanying brain activation (epileptic seizures and direct electrical stimulation) and hypoxia (arterial hypoxia and cyanide poisoning of the brain cortex). In chloralose-anesthetized cats a circumscribed area of the brain cortex was treated with adenosine deaminase (Type III; Sigma), which potently deaminates adenosine to the nonvasoactive inosine. Cerebrocortical vascular volume and fluorescence of reduced nicotinamide adenine dinucleotide were measured in vivo by surface fluororeflectometry. The responses of small pial and intracortical vessels to brain activation and hypoxia were studied in brain cortices superfused with artificial (mock) CSF and 5 U/ml adenosine deaminase. It was found that superficially applied adenosine deaminase readily diffuses onto the brain cortex. Prolonged pretreatment of the brain cortices with 0.025 U/ml adenosine deaminase eliminated almost completely the vasodilative effect of 10(-7) mol/ml adenosine. The inhibitory effect of the enzyme on adenosine-induced cortical vasodilatation was specific, because 5 U/ml adenosine deaminase did not attenuate the vasodilative potency of 10(-8) mol/ml 2-chloroadenosine. Adenosine deaminase (5 U/ml) pretreatment of the brain cortices did not diminish the cerebrocortical vascular volume, which increased with arterial hypoxia, topical cyanide poisoning, and direct electrical stimulation. However, it slightly decreased the vasodilative effect of epileptic seizures. On the basis of these results, it seems very unlikely that adenosine is a critical factor in the control of cerebrovascular tone during arterial hypoxia and brain activation.

Contractile and endothelium-dependent dilatory responses of cerebral arteries at various extracellular magnesium concentrations.

To clarify the effect of extracellular magnesium (Mg2+) on the vascular reactivity of feline isolated middle cerebral arteries, the effects of slight alterations in the Mg2+ concentration on the contractile and endothelium-dependent dilatory responses were investigated in vitro. The contractions, induced by 10(-8)-10(-5) M norepinephrine, were significantly potentiated at low Mg2+ (0.8 mM v. the normal, 1.2 mM). High (1.6 and 2.0 mM) Mg2+ exhibited an inhibitory effect on the contractile responses. No significant changes, however, in the EC50 values for norepinephrine were found. The endothelium-dependent relaxations induced by 10(-8)-10(-5) M acetylcholine were inhibited by high (1.6 and 2.0 mM) Mg2+. Lowering of the Mg2+ concentration to 0.8 mM or total withdrawal of this ion from the medium failed to alter the dilatory potency of acetylcholine. The changes in the dilatory responses also shifted the EC50 values for acetylcholine to the right. The present results show that the contractile responses of the cerebral arteries are extremely susceptible to the changes of Mg2+ concentrations. In response to contractile and endothelium-dependent dilatory agonists, Mg2+ probably affects both the calcium influx into the endothelial and smooth muscle cells as well as the binding of acetylcholine to its endothelial receptor. Since Mg2+ deficiency might facilitate the contractile but not the endothelium-dependent relaxant responses, the present study supports a role for Mg2+ deficiency in the development of the cerebral vasospasm.

Pressure distribution in the pial arterial system of rats based on morphometric data and mathematical models.

The objective of the present work was a theoretical evaluation of pial arterial pressures in normotensive rats and spontaneously hypertensive rats based on the geometry and topography of the pial arterial system as well as on various topological models of the vascular trees. Pial branches of the middle cerebral artery in the diameter range of 30-320 microns were selectively visualized by corrosion compound, and the diameter and length of vascular segments were measured. The vessels were classified into branching orders by the methods of Horsfield and Strahler. The steady-state pressure distribution in the pial arterial system was calculated assuming that the flow at the bifurcations was partitioned in proportion to a given power of the diameters of the daughter branches (diameter exponent). The maximum number of vascular segments along the longest branch varied between 16 and 33. The mean branching ratio was 4.14 +/- 0.23 (SD). The mean diameter of vessels classified into Strahler orders 1-5 were: 50 +/- 12, 71 +/- 19, 106 +/- 22, 168 +/- 22, and 191 +/- 7 microns, respectively. The calculated pressure drop in the pial trees of normotensive rats was approximately twice as large in proximal orders 3 and 4 than in distal orders 1 and 2. The mean pressure in arteries of order 1 ranged from 54.4 to 58.4 mm Hg in the normotensive rat (input pressure: 83 mm Hg), and from 77.2 to 89.0 mm Hg in the spontaneously hypertensive rat (input pressure: 110 mm Hg). The coefficient of variation of terminal pressures in vessels of order 1 increased linearly with the mean pressure drop in the system. The coefficient of variation in terminal pressure had a minimum as a function of the diameter exponent in case of each pial tree. At its minimum, it was higher in all spontaneously hypertensive rats (10.1-22.9%) than in any normotensive rats (6.0-8.5%). The corresponding diameter exponents were in most cases lower in the spontaneously hypertensive rat (1.3-2.5) than in the normotensive rat (2.5-3.0). Topologically consistent models of the pial arterial network predicted significantly less variation in intravascular pressures than was obtained by direct calculations. More idealized models suggested the dependence of coefficient of variation in terminal pressure on the total number of vascular segments contained by the tree. All models predicted the existence of the minimum of coefficient of variation in terminal pressure in function of the diameter exponent.(ABSTRACT TRUNCATED AT 400 WORDS)

Biomechanical properties of normal and fibrosclerotic human cerebral arteries.

Quasi-static passive mechanical properties of histologically fibrosclerotic and normal groups of human anterior cerebral arteries (ACA) and internal carotid arteries (ICA) were studied in vivo. Cylindrical arterial segments were subjected to slow, cyclic inflation by air in the range of 5-250 mm Hg intraluminal pressure at axial isometry. To characterize mechanical properties, incremental elastic modulus, incremental distensibility and strain energy density were computed from the continuously recorded pressure-external diameter curves. Compared to normal arteries, at identical intraluminal pressures, the elastic modulus of fibrosclerotic arteries was found to be 34-45% lower in ACA and 40-56% lower in ICA, and the radius to wall thickness ratio was 25-30% smaller in ACA and 37-38% smaller in ICA. Distensibility of fibrosclerotic arteries was not smaller than that of the normal vessels. There were no significant differences in internal radius and in strain energy density between the fibrosclerotic and normal groups. Results of mathematical modelling suggests that the observed decrease in the elastic modulus of fibrosclerotic arteries was accompanied by different types of structural reorganization in the case of ACA and ICA. It is supposed that changes in mechanical properties of the passive wall elements have a compensatory character to restore some hemodynamically important properties of fibrosclerotic arteries, namely tangential stress, incremental distensibility or characteristic impedance.

Effect of two week lymphatic occlusion on the mechanical properties of dog femoral arteries.

In order to study the long-term effect of impaired lymphatic drainage on the mechanical properties of the arterial wall, cylindrical femoral artery segments from 10 mongrel dogs after 2 weeks of hindlimb lymphatic occlusion were subjected to in vitro mechanical test and compared with the contralateral, sham-operated segments. Smooth muscle contraction was induced by norepinephrine (7.4 X 10(-6) M) and smooth muscle relaxation by papaverine (1.6 X 10(-4) M). As a result of 2 weeks of lymphatic occlusion, wall thickness increased from 243 +/- 18 to 343 +/- 35 microns (P less than 0.02), inner radius decreased from 1.69 +/- 0.11 to 1.42 +/- 0.12 mm (P less than 0.01) and elastic modulus decreased from 1.23 X 10(6) to 0.55 X 10(6) N/m2 (P less than 0.01), when determined at 100 mm Hg (13.3 kPa) intraluminal pressure and with relaxed smooth muscle. The contractile apparatus was able to produce active strain in the vessels with lymphostasis and at physiological pressures not significantly different from the controls (0.89 +/- 0.02 vs. 0.91 +/- 0.02), but at significantly lower levels of tangential stress. Active stress decreased significantly. This study shows that a reorganization of the vessel wall mechanical force-bearing elements occurs in lymphostasis, which, in some respects, resembles the mechanical alterations found in different forms of atherosclerosis.

Prevention of the hemorrhagic hypotension-induced hepatic arterial vasoconstriction by L-arginine and naloxone.

The possible involvement of the L-arginine-nitric oxide pathway and endogenous opioid mechanisms in the hemorrhagic hypotension- (HH) induced changes of hepatic arterial blood flow and vascular resistance was studied in cats. During HH hepatic arterial blood flow was significantly higher both in L-arginine- and naloxone-treated animals than in controls. Furthermore, HH induced a significant increase of the hepatic vascular resistance in the control group, which was prevented by L-arginine or naloxone treatment. Because inhibition of the nitric oxide synthesis by N(G)-nitro-L-arginine in normotensive cats induced a similar increase of the hepatic vascular resistance to that observed during HH in the control group, our results indicate that impairment of the endothelial function may be responsible for the hemorrhage-induced L-arginine- and naloxone-reversible hepatic arterial vasoconstriction. This hypothesis is consistent with our previous observations demonstrating the development of endothelial dysfunction in the feline hepatic artery during HH.

Cerebrocortical and medullary blood flow changes after general opiate receptor blockade during hemorrhagic shock in cats.

The effect of centrally induced opiate receptor blockade on regional cerebral blood flow (rCBF) was studied in anesthetized, ventilated cats during the course of hemorrhagic shock. The blood flow of the medulla and the parietal cortex was measured with the H2-gas clearance technique. Hemorrhagic shock was produced by lowering the systemic mean arterial pressure to 60 mmHg for 120 min by blood withdrawal. Central opiate receptor blockade was induced by 10 micrograms/kg intracerebroventricularly (i.c.v.) injected naloxone at the 60th min of the bleeding period. Cortical blood flow showed no improvement after i.c.v. naloxone administration. Medullary blood flow, however, increased significantly and approached the pre-bleeding control flow values following central opiate receptor blockade. The results indicate involvement of endogenous opioid mechanisms in the regulation of rCBF during hemorrhage and may provide an explanation for the previously described beneficial effects of naloxone in hemorrhagic shock.

Effect of L-arginine on adrenal and renal blood flows during hemorrhage in cats.

Our earlier studies have shown development of endothelial dysfunction in the feline renal artery during hemorrhagic hypotension. Because L-arginine (L-Arg), the precursor of nitric oxide (NO), reportedly improves endothelial function in several pathophysiological states including hypotension, we investigated its possible beneficial effect on the adrenal and renal circulations during hemorrhagic hypotension in anesthetized, ventilated cats. Hypotension (mean arterial pressure 50 mm Hg) significantly increased vascular resistance and decreased blood flow (radiolabeled microspheres) in both adrenal and renal cortices. L-Arg (30 mg/kg bolus, 10 mg/kg/min infusion, i.v.) had no significant hemodynamic effects in normotension but prevented the increase of the vascular resistance and improved blood flow in the adrenal cortex during hypotension. In the kidney, L-Arg also prevented hemorrhage-induced vasoconstriction, although its effect on blood flow did not reach significance. The NO synthase inhibitor N(G)-nitro-L-arginine (30 mg/kg bolus, 1 mg/kg/min infusion, i.v.) increased adrenal and renal vascular resistances to a similar extent as that observed during hypotension. It thus seems that an L-Arg-reversible dysfunction of the endothelial NO-synthesizing pathway contributes to hemorrhage-induced adrenal and renal vasoconstriction.

Determinants of brain activation-induced cortical NAD/NADH responses in vivo.

In order to elucidate that which are the factors that may influence the direction of brain activation-induced changes in the redox state of oxidized/reduced nicotinamide adenine dinucleotide (NAD/NADH), the brain cortex was electrically stimulated during arterial hypotension and following reinfusion of the shed blood, during arterial hyper- and hypoxia, and during the second phase of spreading cortical depression (SD). Cerebrocortical NADH fluorescence and vascular volume ( CVV ) of cats, anaesthetized by chloralose, were measured with a microscope fluororeflectometer . Under physiologically normal conditions electrical stimulation resulted in pronounced cortical NAD reduction and increase in CVV . These reactions were not altered by arterial hyperoxia and continuous superfusion of the brain cortex with oxygenated artificial cerebrospinal fluid (mock CSF). Arterial hypotension and SD (in phase II) increased NAD reduction and CVV markedly, and the superimposed electrical stimulation brought about NADH oxidation and greatly depressed CVV responses. Reinfusion of the shed blood did not restore NAD/NADH redox state and CVV to their reference levels, and electrical stimulation under this condition led to NADH oxidation and negligible vascular reactions. Since under physiologically normal conditions electrical activation of the brain cortex resulted in NAD reduction and marked increase in CVV and the magnitude of these reactions were not altered by arterial hyperoxia or by superfusion of the brain cortex with oxygenated CSF, it is very unlikely that the brain cortex became hypoxic during stimulation. Because when the steady NAD/NADH redox state of the brain cortex was shifted toward reduction by arterial hypotension and reinfusion and SD, electrical stimulation led to NADH oxidation, it is suggested that the prestimulatory steady redox state has great importance in determining the direction of NAD/NADH redox reactions evoked by activation of the brain cortex.

Role of platelet-activating factor in the development of endothelial dysfunction in hemorrhagic hypotension and retransfusion.

Platelet-activating factor (PAF), an important mediator of ischemic and shock states, has been shown to prime direct and neutrophil-mediated endothelial cell injury. In the present study we investigated therefore whether PAF is involved in the development of dysfunction of the cerebrovascular endothelium in hemorrhagic hypotension and retransfusion in cats. In vitro responses of middle cerebral arteries prepared from control animals and from animals subjected to hemorrhagic hypotension with or without specific PAF antagonist WEB 2086 treatment (1 mg/kg initial bolus followed by a 0.05 mg/kg/min infusion) were studied by measuring isometric force in organ chambers containing Krebs-Henseleit solution (37 degrees C, gassed with 95% O2-5% CO2). Bleeding was performed in a stepwise fashion by bringing the mean arterial blood pressure to 90, 70 and 50 mmHg and maintained for 20 min at each level followed by a 20-min retransfusion. Hemorrhagic hypotension and retransfusion caused a marked attenuation of the acetylcholine- and ATP-induced endothelium-dependent relaxations of the middle cerebral artery whereas the dilations induced by the nitric oxide donor and direct vasorelaxant SIN-1, remained unaltered. In the vessels, prepared from animals which received WEB 2086 treatment during hemorrhage and retransfusion there were more pronounced cholinergic (but not purinergic) relaxations than in the untreated animals subjected to hemorrhage. SIN-1 induced relaxations remained unaltered after WEB 2086 treatment. Our results suggest that platelet-activating factor is in part involved in the pathophysiological processes leading to the development of the endothelial dysfunction in the present model of hemorrhagic hypotension and retransfusion.

Evidence for the role of nitric oxide in the circulation of the dental pulp.

Many authors have studied the hemodynamics of the dental pulp; however, there are scarcely any data regarding the involvement of the L-arginine/nitric oxide pathway in the regulatory mechanism. Thus, we have examined the physiological effects of (1) NG-nitro-L-arginine as an inhibitor of nitric oxide synthesis and (2) the nitric oxide donor 3-morpholinosydnonimine on blood flow and vascular resistance in the canines of anesthetized cats to study the potential involvement of nitric oxide in the regulation of dental vascular homeostasis. Mean arterial blood pressure, heart rate, blood gases, pH, cardiac output, and tissue blood flow were determined prior to and 15 min after i.v. administration of either NG-nitro-L-arginine (30 mg/kg, n = 9) or 3-morpholinosydnonimine (1 mg/kg, n = 7). Blood flow was measured by radioactive-labeled microspheres. There were no significant differences in baseline parameters between the two groups of cats. The dental pulp blood flow decreased to 53 +/- 13% (p < 0.01) of the control level after NG-nitro-L-arginine administration, while it decreased only slightly (to 82 +/- 12%) after 3-morpholinosydnonimine administration. The dental pulp's vascular resistance increased to 367 +/- 69% (p < 0.01) of the control level after NG-nitro-L-arginine, while it decreased to 73 +/- 10% (p < 0.05) of control after 3-morpholinosydnonimine. We found that the L-arginine/nitric oxide pathway plays an important role in the regulation of pulpal blood circulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Activated neutrophils inhibit cerebrovascular endothelium-dependent relaxations in vitro.

The effect of formyl-Met-Leu-Phe- (fMLP-) activated feline neutrophil granulocytes on endothelium-dependent and independent relaxations was studied in the middle cerebral artery of the cat in vitro. Endothelium-dependent relaxations caused by acetylcholine and ATP were markedly inhibited after 30 minutes of incubation of the vessels with neutrophils (5000 cells/microliter) in the presence of 5 microM fMLP, followed by a replacement of the bath solution in order to remove the neutrophils from the medium. Direct vasorelaxations in response to the nitric oxide donor compound SIN-1, however, remained unchanged. Both neutrophils and fMLP caused transient contractions during the incubation period. The present study provides direct evidence for the ability of activated neutrophils to cause an inhibition of vascular endothelium-dependent responses in vitro.

The effect of a nitric oxide donor and an inhibitor of nitric oxide synthase on blood flow and vascular resistance in feline submandibular, parotid and pancreatic glands.

The aim was to examine whether (1) blood flow and vascular resistance are altered in response to exogenous nitric oxide and (2) whether endogenous synthesis of nitric oxide participates in the haemodynamic regulation of the submandibular, parotid and pancreatic glands. Experiments were performed on anaesthetized, artificially ventilated cats. Mean arterial blood pressure, heart rate, blood gases, cardiac output and tissue blood flow were determined before and 15 min after intravenous administration of either the nitric oxide donor SIN-1 (3-morpholinosydnonimine, 1 mg/kg, n = 10) or the competitive nitric oxide synthase inhibitor NOLA (NG-nitro-L-arginine, 30 mg/kg, n = 9) blood flow was measured by a radioactive-labelled microsphere method. In the SIN-1 group, in spite of a serious decrease in mean arterial blood pressure (p < 0.001), the blood flow in the glands remained unchanged. The vascular resistance decreased after SIN-1 in the submandibular and pancreatic glands (p < 0.001 and p < 0.05, respectively), and was slightly reduced in the parotid. The NOLA increased mean arterial blood pressure (p < 0.01) and reduced the blood flow in the submandibular and pancreatic glands (p < 0.01 and p < 0.001, respectively), but the decrease in the parotid was not significant. Vascular resistance increased after NOLA in all three glands (p < 0.05, p < 0.001 and p < 0.05). These findings suggest that basal nitric oxide production in these exocrine glands is sufficient to modulate vascular resistance. Moreover, the release of endogenous NO from the nerves and/or endothelium is probably involved in the regulation of vascular tone. The nitric oxide-dependent component of blood-flow regulation, however, seems to be less pronounced in the parotid gland.

Regulation of cerebral blood flow (CBF) during hypoxia and epileptic seizures.

In conclusion, our results suggest that neither the redox state of cytochrome oxidase nor adenosine are critical factors in the regulation of cerebral blood flow during arterial hypoxia and epileptic seizures.

Transient metabolic and vascular volume changes following rapid blood pressure alterations which precede the autoregulatory vasodilation of cerebrocortical vessels.

It has been shown by surface fluoro-reflectometry that stepwise decrease of arterial blood pressure causes a biphasic cerebrocortical vascular volume response. After the arterial blood pressure decrease the vascular volume first decreased and later increased. In both parts of the biphasic reflectance change, the cerebrocortical NAD-NADH redox state shifted considerably towards reduction and there was no reoxidation after the onset of cortical vasodilatation. Since a very rapid NADH reduction occurred during the first 30 secs. of the arterial hypotension in parallel with the vascular volume decrease, it is suggested that in the transient phase of arterial hypotension cerebral hypoxia may occur. Furthermore it is suggested that anaerobic tissue metabolites or some unknown NAD-NADH dependent process might dilate the cerebrocortical arterial network during the autoregulatory adjustment of CBF. The participation of the sympathetico-adrenal system in transient brain hypoxia caused by bleeding is a possibility since both the early vasoconstriction and the steep NADH reduction were prevented by the administration of phenoxybenzamine (1 mg/kg) before bleeding.

Metabolic and electron microscopic studies post mortem in brain mitochondria.

The RCR of malate+glutamate oxidation of isolated rat brain mitochondria decreased with increasing duration of anoxia and it ceased completely in the 12 hour ischemic group. After one hour of ischemic anoxia neither the respiration of mitochondria nor their morphology altered irreversibly. There is a strong correlation between the ultrastructural changes and RCR value of mitochondria, namely the RCR decreased proportionally with increasing number of light type of mitochondria. From our results we can conclude that the dense form seems to be the functional form, while the light mitochondrial structure is a transition to the non functioning homogenous type of brain mitochondria.