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Biomedical subjects

R E Purdy

Publications and source records attributed to R E Purdy.

88 records · Page 5Linked to original sources

Changes in the contractile response of arteires and veins from hypertensive rabbits to sympathetic nerve activity: assessment of some postsynaptic influences.

Contractile responses to field stimulation of intramural nerves of arteries and veins taken from rabbits made hypertensive by partial constriction of the abdominal aorta have been related to the carotid artery pressure. The increase in contraction of cephalic and short saphenous veins with rise in carotid artery pressure can be accounted for by an increase in sensitivity of the alpha-adrenergic receptor. The neurogenic contraction of the ear artery increased with carotid artery pressure rise. Changes in some of the extraneuronal factors that influence transmitter distribution and disposition in the tunica media were examined. In hypertensive animals, the percentage of released adrenergic transmitter entering the vessel wall might be expected to decrease due to an increase in medial thickness. However, this percentage was not significantly altered in the ear artery probably due, in part, to a concomitant increase in medial permeability to the transmitter. Extraneuronal transmitter disposition factors, i.e. extraneuronal uptake, monoamine oxidase, and catechol-O-methyltransferase activity are directly related to the wet weight of the vessel wall. Thus, their contribution to transmitter disposition would be expected to increase with increase in vessel wall thickness and tend to reduce the response to sympathetic activity. As the contractile response increased in the hypertensive vessels despite such changes, the increase in effector cell mass and density of neuronal terminal plexus, shown previously to increase with hypertension, are more important than these other considerations.

Animals↗

Hydrolysis of plant cuticle by plant pathogens. Purification, amino acid composition, and molecular weight of two isozymes of cutinase and a nonspecific esterase from Fusarium solani f. pisi.

The extracellular fluid of the plant pathogen, Fusarium solani f. pisi, grown on the plant cuticular polymer, cutin, was shown to contain cutinase and p-nitrophenyl palmitate hydrolase activities (R.E. Purdy and P.E. Kolattukudy (1973), Arch. Biochem. Biophys. 159, 61). From this extracellular fluid two isozymes of cutinase and a nonspecific esterase (p-nitrophenyl palmitate hydrolase) were isolated using Sephedex G-100 gel filtration, QAE-Sephadex chromatography, and SE-Sephedex chromatography. Phenolics contained in the extracellular fluid were found to be associated with the cutinase but not with the nonspecific esterase, and the phenolic materials were removed from cutinase at the QAE-Sephedex step. A 34-fold purification of the nonspecific esterase and a 6.5-fold purification of cutinase were achieved by the procedure described. The two isozymes of cutinase (I and II) and the nonspecific esterase were homogeneous as judged by polyacrylamide disc gel electrophoresis and sedimentation equilibrium centrifugation. Molecular weights of cutinase I, cutinase II, and the nonspecific esterase were determined by Sephedex G-100 gel filtration, sedimentation equilibrium centrifugation, amino acid composition, and sodium dodecyl sulfate polyacrylamide disc gel electrophoresis. The values obtained with these techniques agreed with each other and were about 22,000 for both cutinases and 52,000 for the nonspecific esterase. The dodecyl sulfate gel electrophoresis indicated that a small portion of cutinase II contained proteolylic clips, near the middle of the polypeptide chain, and that the nonspecific esterase might also have undergone some proteolylic modification. The amino acid composition of cutinase I was similar to that of cutinase II except for the presence of a larger number of tryptophan residues in the latter, while the amino acid composition of the nonspecific esterase showed more differences from that of either cutinase.

Amino Acids↗

Hydrolysis of plant cuticle by plant pathogens. Properties of cutinase I, cutinase II, and a nonspecific esterase isolated from Fusarium solani pisi.

The properties of the homogeneous cutinase I, cutinase II, and the nonspecific esterase isolated from the extracellular fluid of cutin-grown Fusarium solani F. pisi (R.E. Purdy and P.E. Kolattukudy (1975), Biochemistry, preceding paper in this issue) were investigated. Using tritiated apple cutin as substrate, the two cutinases showed similar substrate concentration dependence, protein concentration dependence, time course profiles, and pH dependence profiles with optimum near 10.0. Using unlabeled cutin, the rate of dihydroxyhexadecanoic acid release from apple fruit cutin by cutinase I was determined to be 4.4 mumol per min per mg. The cutinases hydrolyzed methyl hexadecanoate, cyclohexyl hexadecanoate, and to a much lesser extent hexadecyl hexadecanoate but not 9-hexadecanoyloxyheptadecane, cholesteryl hexadecanoate, or hexadecyl cinnamate. The extent of hydrolysis of these model substrates by cutinase I was at least three times that by cutinase II. The nonspecific esterase hydrolyzed all of the above esters except hexadecyl cinnamate, and did so to a much greater extent than did the cutinases. None of the enzymes hydrolyzed alpha- or beta-glucosides of p-nitrophenol. p-Nitrophenyl esters of fatty acids from C2 through C18 were used as substrates and V's and Kms were determined...

Binding Sites↗

Vasomotor nerve control of isolated arteries and veins.

In order to compare neuro-effector function in different blood vessels, frequency-response relationships were determined for the following preparations: 1)Isolated rings of the proximal saphenous, distal saphenous and ear arteries, the parietal branch of the internal iliac vein and the small saphenous vein from the rabbit, 2) spiral strips of the rabbit pulmonary artery and 3) longitudinal preparations of the rat portal vein. In each rabbit tissue only one low (less than or equal to 4 Hz) and one high (larger than or equal to 8 Hz) transmural nerve stimulation frequency was applied until steady state responses were obtained and these were expressed as a percentage of a maximum response to exogenous noradrenaline (NA) applied in each experiment. The general shape of the frequency-response curves was similar, but differences in steepness and amplitude of the maximum neurogenic response relative to exogenous NA were found. The steepness of the frequency-response relationships of the veins tended to be greater than those of the arteries. It appears that factors such as close neuro-muscular contacts, presence of terminal nerve fibres within the media and the operation of mechanisms for myogenic propagation of activity contribute to the effectiveness of neurogenic vascular control as revealed by frequency-response curves. In vivo, geometrical factors can greatly augment the hemodynamic significance of the observed differences.

Animals↗

Analysis of changes in reactivity of rabbit arteries and veins two weeks after induction of hypertension by coarctation of the abdominal aorta.

Vessel dimensions and characteristic responses to norepinephrine were measured in various arteries and veins of the rabbit made hypertensive by partial constriction of the upper abdominal aorta. The ear, radial, and basilar arteries taken from the circulation proximal to the ligature (the hypertensive arteries) were thickened in proportion to the rise is arterial blood pressure. The water, sodium, and potassium contents of these and all other vessels were not significantly changed in the hypertensive rabbits. The maximum response to norepinephrine in the ear artery, a representative vessel from the hypertensive part of the rabbit, was increased, whhereas the sensitivity of this vessel to norepinephrine expressed as the ED50 did not alter with changes in the arterial blood pressure. In contrast, the thickness and the maximum response to norepinephrine of the saphenous artery, representative of vessels distal to the ligature (normotensive vessels) and of the saphenous and cephalic veins were unaltered. The sensitivity as indicated by the norepinephrine ED50 of the veins, but not of the saphenous artery, increased with a rise in carotid artery blood pressure. These results suggest that the increased responsiveness to norepinephrine of arteries proximal to the ligature is due to changes in muscle mass and that the increased responsiveness of the veins is due to increased sensitivity to norepinephrine.

Animals↗

Evidence for an increase in adrenergic nerve function in blood vessels from experimental hypertensive rabbits.

The possibility of changes in the adrenergic innervation of blood vessels in experimental hypertension was investigated by measuring arterial norepinephrine content, neuronal uptake of norepinephrine, and the neurogenic contractile response in rabbits made hypertensive by partial constriction of the abdominal aorta proximal to the kidneys. Two to 3 weeks after surgery, norepinephrine content was increased in the arteries above the ligature, where arterial blood pressure was increased, but not in the arteries below the ligature, where arterial blood pressure was normal, in the heart, or in the veins. Neuronal norepinephrine uptake per unit length of vessel and the neurogenic contractile response increased with the rise in arterial blood pressure. The neurogenic contractile response can be taken as an indication of an increase in transmitter release. The results taken together suggest an increase in the function and possibly the amount of the adrenergic neuroneal terminal in hypertension. Since the distributions of the changes in the adrenergic innervation and the increases in smooth muscle cell proliferation in hypertension are similar, these two processes may be interrelated.

Animals↗

Cardiac function after eight hour storage by using polyethylene glycol hemoglobin versus crystalloid perfusion.

Efforts to extend myocardial preservation for transplantation by crystalloid perfusion have been limited by edema and compromised function. We hypothesized that hypothermic perfusion preservation with a polyethylene glycol (PEG) conjugated hemoglobin solution may extend preservation times. The purpose of this study was to compare cardiac function after continuous perfusion by using a hypocalcemic, normokalemic crystalloid perfusate with and without the addition of PEG-hemoglobin (Hb). The hearts of 20 anesthetized and ventilated New Zealand White rabbits were harvested after cold cardioplegic arrest. Group I (n = 10) hearts were continuously perfused with a hypocalcemic, normokalemic 3% bovine PEG-Hb solution at 20 degrees C and 30 mm Hg for 8 hours. Group II (n = 10) hearts were continuously perfused with an identical crystalloid solution without PEG-Hb for 8 hours under the same conditions as group I hearts. Cardiac function was measured with a left ventricular force transducer after transfer to a standard crystalloid Langendorff circuit at 37 degrees C and an aortic root pressure of 59 mm Hg. After 8 hours of perfusion preservation, heart rate was similar for groups I and II (p = not significant [NS]). Coronary blood flow after and during preservation was similar between PEG-Hb and crystalloid preserved hearts (p = NS). Left ventricular developed pressure, peak dP/dt, and peak -dP/dt were superior in hearts preserved with PEG-Hb. Percent water of total ventricular weight was 82.0% for group I and 81.6% for group II (p = NS). Continuous perfusion preservation of rabbit hearts for 8 hours with a hypocalcemic normokalemic PEG-Hb based solution at 30 mm Hg and 20 degrees C yields left ventricular function that is superior to perfusion with a similar crystalloid solution without PEG-Hb, despite similar myocardial edema and coronary flow. Extended cardiac perfusion preservation with this PEG-Hb based solution deserves further study, including comparison with traditional cardioplegic preservation solutions.

Animals↗

Recovery of cardiac function after standard hypothermic storage versus preservation with Peg-hemoglobin.

Preservation of the heart for transplantation after infusion of cardioplegia and extirpation of a cardiac allograft results in an ischemic insult to the myocardium. This ischemic insult may lead to a loss of function in the transplanted heart. Hypothermic perfusion preservation with an oxygen hemoglobin carrying solution may avert ischemic injury and lead to improved recovery of cardiac function. The purpose of this study was to compare cardiac function after 8 hours of continuous hypothermic perfusion with a unique polyethylene-glycol-hemoglobin (PEG-Hb) solution to hearts preserved by 4 hours of hypothermic ischemic storage. Freshly extirpated hearts served as functional controls. The hearts of 26 anesthetized and intubated New Zealand white rabbits were harvested after cold cardioplegic arrest. Group I (n = 12) hearts were perfused with a PEG-Hb solution at 20 degrees C and 30 mm Hg for 8 hours. PO2 was maintained > or = 500 mm Hg. Group II (n = 7) hearts were preserved by cold ischemic storage for 4 hours at 4 degrees C. Group III (n = 7) were tested immediately after harvest. Left ventricular (LV) function was measured in the nonworking state at 15 minutes, 1 hour, and 2 hours after transfer to a standard crystalloid Langendorff circuit. Measurement of LV developed pressure, peak + dP/dt and -dP/dt revealed a superior trend between Group I and Group II hearts in comparison with freshly extirpated hearts. Heart rate was similar among all groups throughout testing (p = ns). Coronary blood flow was not significantly different between groups. Continuous perfusion preservation of rabbit hearts for 8 hours with PEG-Hb solution at 30 mm Hg and 20 degrees C yielded LV function that was similar to 4 hours of ischemic hypothermic storage. Furthermore, return of cardiac function after 8 hours of perfusion preservation using this PEG-Hb solution may be superior to that obtained in freshly extirpated hearts. These data suggest that some recovery of myocardial function may occur during perfusion preservation with this PEG-Hb solution after the ischemic insult of cardioplegic arrest. Continuous perfusion preservation using this PEG-Hb solution deserves further investigation in large animal transplant models.

Animals↗

Adrenergic innervation of large cerebral blood vessels of the rabbit studied by fluorescence microscopy. Absence of features that might contribute to non-uniform change in cerebral blood flow.

The literature provides evidence for non-uniform regional changes in cerebral blood flow under a variety of circumstances. Possible causes of such changes were sought in the larger cerebral arteries of the rabbit prepared according to the glyoxylic acid fluorescence histochemical technique. The adrenergic innervation of the circle of Willis, the basilar artery and their main branches, although showing small differences, is essentially of uniform density. There is no evidence for collars of adrenergic nerves around the origin of small branches nor for cushions at their orifices. Innervation density appears to diminish as the pial vessels get smaller and all vessels seem to be innervated. Thus non-uniform alterations of cerebral blood flow cannot be accounted for by these factors.

Adrenergic Fibers↗

Summary of studies of changes in vascular reactivity caused by natriuretic hormones.

Endogenous Na, K pump inhibitors may contribute to the pathogenesis of hypertension, and could do so by causing direct vasoconstriction and/or enhancing sensitivity to other vasoconstrictor agents. These effects of the Na, K pump inhibitors are likely due to inhibition of Na-K-ATPase. In turn, cells become depolarized, internal sodium concentration increases and internal calcium is increased by exchange for sodium via the sodium/calcium exchange carrier. This extra calcium is sequestered, increasing the size of the releasable intracellular calcium pool. Both depolarization and the increase in cytosolic calcium can cause vasoconstriction. Both depolarization and the increased size the intracellular calcium pool can sensitize the blood vessel to other vasoconstrictor agents. Endogenous pump inhibitors may also stimulate the release of catecholamines from the intramural sympathetic nerve terminals. Studies of a variety of candidate endogenous Na, K pump inhibitors are reviewed. These include presently unidentified substances extracted from human urine, from peritoneal dialysate of hypertensives with renal failure, and from bovine and rat hypothalamus. Additional candidate compounds include ouabain, selected pregnanes and marinobufagenin, a steroid originally identified in the venom of the frog, Bufo marinus.

Animals↗