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J L Gordon

Publications and source records attributed to J L Gordon.

At least 37 records · Page 2Linked to original sources
PubMed1988

ADP plays a key role in thrombogenesis in rats.

The relative importance of ADP, arachidonic acid metabolites and serotonin as thrombogenic factors was evaluated in rats by comparing, after oral administration, the effects of two inhibitors of ADP-induced platelet aggregation (ticlopidine and PCR 4099), three cyclo-oxygenase inhibitors (aspirin, triflusal and indobufen) and a selective serotonin 5HT2 receptor antagonist (ketanserin) on platelet aggregation, in four platelet-dependent thrombosis models and on bleeding time. Platelet aggregation induced by ADP and collagen was completely inhibited by ticlopidine and PCR 4099 whereas only the collagen aggregation was reduced by the cyclo-oxygenase inhibitors. Ketanserin or a depletion of platelet serotonin by reserpine did not affect platelet aggregation. Ticlopidine and PCR 4099 greatly prolonged rat tail transection bleeding time. This is probably related to their known ability to inhibit ADP-mediated platelet aggregation. In contrast, the cyclooxygenase inhibitors did not affect bleeding time at all. Reserpine and ketanserin prolonged bleeding time by interfering with the action of serotonin on the vascular wall. Ticlopidine and PCR 4099 were very potent antithrombotics in all the models. Aspirin, only at a high dose, inhibited poorly thrombus formation on a silk thread in an arterio-venous shunt, suggesting that the inhibition of cyclo-oxygenase was not responsible. Triflusal was inactive in all models while indobufen slightly reduced thrombus formation in the silk thread and metallic coil models. Ketanserin and reserpine reduced thrombus only in the metallic coil model. Thrombus formation was greatly reduced in fawn-hooded rats, which lack ADP in their platelet dense granules because of a genetic storage pool deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Diphosphate↗
PubMed1988

Endothelial functional responses and increased vascular permeability induced by polycations.

Polycations such as poly-L-lysine powerfully stimulated cultured endothelial cells from pig aorta to release prostacyclin and cytoplasmic purines in a dose (charge)-dependent and molecular weight (size) dependent manner. Neutral or anionic polymers were inactive. Qualitatively similar findings were made in vivo where poly-L-lysine induced charge and size-dependent local edema formation after intradermal injection in the rabbit. Pretreatment of cultured endothelium with heparin or trypsin (but not neuraminidase) effectively reduced the response of the cells to subsequent exposure to poly-L-lysine suggesting an interaction of polycations with integral membrane proteins. Edema responses to poly-L-lysine were reduced in the presence of indomethacin suggesting that generation of an endogenous vasodilator prostaglandin, perhaps endothelial cell-derived, was an important component of the response. Poly-L-lysine-induced edema formation was not dependent on endogenous histamine release but was reduced by locally administered trasylol while soybean trypsin inhibitor failed to inhibit the response. Our results indicate that polycations such as poly-L-lysine can induce responses of vascular endothelium in vitro and in vivo and that the effects are not only charge-related but are also dependent on the size of the polycation. We suggest that naturally occurring polycations such as those derived from leukocytes and platelets may play an important role in various pathologic processes and that this may be closely related to their size.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗
PubMed1987

Inhibition of vascular smooth muscle cell proliferation in culture by pentosan polysulphate and related compounds.

Pentosan polysulphate (PPS; SP 54; Hemoclar), a highly sulphated semi-synthetic polysaccharide of MW 4.7 kD, was tested in vitro as an inhibitor of rabbit aortic smooth muscle cell (SMC) proliferation and its effects were compared with those of dextran sulphate, laminarin sulphate and heparin fractions. When added to the cell cultures simultaneously with foetal calf serum, all sulphated polysaccharides inhibited cell growth. PPS was five fold more active than heparin and its low molecular weight fractions. A high molecular weight fraction of PPS was the most active inhibitor (IC50: 2.7 micrograms/ml vs 45 micrograms/ml for PPS, and 241 micrograms/ml for heparin). When PPS and heparin were pre-incubated with the cells for three days, then removed before foetal calf serum was added to stimulate growth, their inhibitory effects were comparable with those seen when the compounds were present throughout the growth phase, suggesting that they exert their effect by binding to the cell surface, and not by interacting with the growth factors in serum.

Animals↗
PubMed1987

Characteristics of the P2 purinoceptor that mediates prostacyclin production by pig aortic endothelial cells.

Release of prostacyclin was studied by superfusing small columns containing cells cultured on microcarrier beads. Transient dose-dependent stimulation of prostacyclin release by up to 500-fold was induced by adenosine 5'-triphosphate (ATP; 0.5-50 microM). Adenosine 5'-diphosphate (ADP) gave similar responses, whereas adenosine 5'-phosphate (AMP) and adenosine were essentially inactive. Of other natural nucleotides tested only uridine 5'-phosphate (UTP) was active. The L-enantiomers of ATP and ADP were inactive. 2-Cl-ATP was approximately 100 times more potent than ATP; 2-MeS-ATP was also more potent (threshold 0.05 microM) but its maximal effectiveness was less than 20% that of ATP; 2-EtS-ATP had a similar threshold to ATP but was even less effective than 2-MeS-ATP. Phosphorothioate nucleotide analogues of ATP or ADP were active, with no stereoselectivity between Rp and Sp diastereoisomers. No analogue tested showed antagonist activity. We conclude that ATP mediates endothelial prostacyclin release apparently via a P2Y receptor, although there are some striking differences from the previously described P2Y receptor mediating endothelium-dependent vasodilation in pig aorta.

Adenosine Triphosphate↗
PubMed1987

Purinoceptors in the rat heart.

The effects of an intracoronary bolus of adenosine triphosphate (ATP), alpha, beta-methylene ATP (APCPP), beta, gamma-methylene ATP (APPCP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and adenosine on coronary tone and ventricular myocardial contraction were investigated in the perfused rat heart. Adenine nucleotides, given by bolus injection were negatively inotropic in amounts greater than 3 X 10(-7) mol. The potency order was ATP greater than ADP greater than AMP. Adenosine (less than 1 X 10(-5)mol) had no effect on ventricular myocardial contraction. Adenine nucleotides and adenosine (1 X 10(-10)-1 X 10(-7) mol) reduced coronary tone. The potency order was ATP greater than ADP greater than AMP = adenosine. The ATP analogue APPCP was less active than ATP at reducing coronary tone, and APCPP had no vasodilator effect. This suggests the presence of a P2-purinoceptor, subclass P2Y, which mediates vasodilation. ATP and ADP increased the concentration of prostacyclin (measured as 6-keto prostaglandin F1 alpha) in the perfusate, but only after injection of greater than 3 X 10(-7) mol, suggesting that the vasodilator responses to ATP and ADP were not mediated by prostacyclin. AMP and adenosine had no effect, even at 1 X 10(-5) mol. At a dose of 3 X 10(-9) mol, approximately 40% of ATP and 70% of ADP was converted to AMP and adenosine whilst passing through the heart. The amounts of AMP and adenosine formed, however, were insufficient to account for the vasodilator effects of ATP and ADP. 6 Vasodilatation mediated by AMP and adenosine was inhibited by an infusion of 8-phenyltheophylline (8-PT; 2 x 10-5 M) indicating interaction with a P1-purinoceptor. Vasodilatation induced by ATP (at doses at which AMP and adenosine had no action) was also depressed by 8-PT indicating either an action of ATP on PI-purinoceptors, or an effect of 8-PT on P2y receptors. 7 Vasodilatation induced by AMP was unaltered during an infusion of alpha,beta-methylene ADP (2 x 10-6 M, which inhibited breakdown of AMP to adenosine by 54.2 +/- 1.5%, n = 4). This suggests that AMP acted directly, and it did not require conversion to adenosine to induce vasodilatation. 8 The ATP analogues APCPP (1 x 10-9_1 x 10-8 mol) and APPCP (1 x 10-8_l x 10-7mol) increased coronary tone, as did high doses (I x 10-5 mol) ofATP and ADP, indicating the presence of an additional P2-purinoceptor, subclass P2X, mediating vasoconstriction.

6-Ketoprostaglandin F1 alpha↗
PubMed1986

A model of the smooth pursuit eye movement system.

Human, horizontal, smooth-pursuit eye movements were recorded by the search coil method in response to Rashbass step-ramp stimuli of 5 to 30 deg/s. Eye velocity records were analyzed by measuring features such as the time, velocity and acceleration of the point of peak acceleration, the time and velocity of the peaks and troughs of ringing and steady-state velocity. These values were averaged and mean responses reconstructed. Three normal subjects were studied and their responses averaged. All showed a peak acceleration-velocity saturation. All had ringing frequencies near 3.8 Hz and the mean steady-state gain was 0.95. It is argued that a single, linear forward path with any transfer function G(s) and a 100 ms delay (latency) cannot simultaneously simulate the initial rise of acceleration and ring at 3.8 Hz based on a Bode analysis. Also such a simple negative feedback model cannot have a steady-state gain greater than 1.0; a situation that occurs frequently experimentally. L.R. Young's model, which employs internal positive feedback to eliminate the built-in unity negative feedback, was felt necessary to resolve this problem and a modification of that model is proposed which simulates the data base. Acceleration saturation is achieved by borrowing the idea of the local feedback model for saccades so that one nonlinearity can account for the acceleration-velocity saturation: the main sequence for pursuit. Motor plasticity or motor learning, recently demonstrated for pursuit, is also incorporated and simulated. It was noticed that the offset of pursuit did not show the ringing seen in the onset so this was quantified in one subject. Offset velocity could be characterized by a single exponential with a time constant of about 90 ms. This observation suggests that fixation is not pursuit at zero velocity and that the pursuit system is turned on when needed and off during fixation.

Eye Movements↗
PubMed1985

Specificity of P2-purinoceptor that mediates endothelium-dependent relaxation of the pig aorta.

ATP (EC50 5 microM) induced endothelium-dependent relaxation of the isolated aorta of the newborn pig, but the other naturally occurring nucleotides CTP, GTP, ITP and UTP were more than 100 times less potent. 2-Methylthio-ATP (EC50 0.1 microM) was 50 times more potent than ATP, but the unnatural enantiomers L-ATP and 2-methylthio-L-ATP were virtually inactive. beta,gamma-Imido-ATP and beta,gamma-methylene-ATP, both of which are resistant to degradation by ectonucleotidases on cultured pig endothelial cells, were much less potent than ATP. ADP beta S, which is also resistant to degradation, was equipotent with ATP at low concentrations but achieved a maximal relaxation of only 50% that of ATP. The Rp and Sp diastereoisomers of ATP beta S were both equipotent with ATP at low concentrations and both achieved approximately 60% of the maximal relaxation of ATP. The Rp and Sp diastereoisomers of ADP alpha S were both less potent than ATP and achieved only approximately 25% of the maximal relaxation of ATP. These results demonstrate that the P2-purinoceptor mediating endothelium-dependent relaxation of the pig aorta exhibits a high degree of specificity for the adenine base, is stereospecific for the D-ribofuranosyl moiety, requires a phosphate chain of 2 or 3 units but is not stereoselective toward this phosphate chain. These structural requirements have some features in common with the P2-purinoceptors on smooth muscle and on platelets, and are quite different from those of the ectonucleotidases present on pig endothelial cells in culture.

Adenosine Triphosphate↗
PubMed1985

Endothelium as a modulator of platelet reactivity.

The pathways through which endothelial cells can modulate platelet reactivity (whether directly via the secretion of agents that affect platelet function or indirectly through the regulation of coagulation or vascular tone) are sufficiently varied and complex that the direction of individual reactions can be altered as the homeostatic balance requires--in other words, the balance can be shifted in favour of activation or inhibition depending on circumstances. For example, endothelium has the capacity to express pro- or anticoagulant activities, and to release or inactivate agents that either promote or inhibit platelet aggregation. The role of endothelial cells in modulating platelet function is important, complex and as yet poorly understood, but our understanding of the processes involved has advanced greatly over the past few years and continues to increase.

Adenosine Diphosphate↗
PubMed1985

Purines and prostaglandins as regulators in the coronary circulation.

In the isolated rat heart perfused at constant flow a bolus injection of an adenine nucleotide reduced coronary perfusion pressure and was negatively inotropic (ATP greater than ADP greater than AMP). Adenosine had no effect on perfusion pressure or contractile force. ATP and ADP stimulated production of PGI2 from the coronary bed. Perfusion with indomethacin completely inhibited PGI2 production but had no effect on the reduction in perfusion pressure and cardiac function produced by ATP. Purinoceptors in the perfused rat heart are of the P2 type and responses produced by purinoceptor activation are not mediated by PGI2.

Adenine Nucleotides↗
PubMed1984

Stimulation of endothelial cells by protease activity in commercial preparations of xanthine oxidase.

The oxygen radical generating system of xanthine oxidase plus xanthine, which has been used as a model for the oxidative burst of activated granulocytes, is known to damage endothelium in vivo and in vitro. We previously observed effects (inhibited by catalase, and thus associated with the formation of H2O2) on several parameters of endothelial function, using a non-commercial preparation of xanthine oxidase. Our present study demonstrates that xanthine oxidase from two different commercial sources has additional effects on endothelial morphology and ion flux that are substrate-independent (i.e. produced in the absence of added xanthine) and are attributable to the presence of pancreatin (a crude enzyme mixture used in the commercial preparation of xanthine oxidase from milk). These effects are related to the tryptic activity of pancreatin and extend previous observations on the effects of neutral proteases on endothelial cells. Our results emphasise the practical point that studies on the effects of commercial xanthine oxidase preparations on endothelial cells must take account of their trypsin-like activity as well as their capacity to generate oxygen products.

Adenine↗
PubMed1984

Differential effects of hydrogen peroxide on indices of endothelial cell function.

The responses of pig aortic endothelial cells to sublethal doses of potentially toxic stimuli were investigated by monitoring K+ efflux, prostaglandin production, and the release of cytoplasmic purines. Xanthine plus xanthine oxidase reversibly stimulated these three parameters of endothelial cell function at doses that were not cytotoxic, as measured by chromium release, adenine uptake, and vital dye exclusion. The effects of xanthine plus xanthine oxidase were inhibited by catalase but not by superoxide dismutase, suggesting that H2O2 was responsible. Reagent H2O2 also reversibly stimulated K+ efflux, prostaglandin production, and the release of purines. The threshold concentration of H2O2 for these effects was approximately 10 microM, which was at least 30-fold lower than that which caused cytotoxicity. In addition to the direct effect of H2O2 in stimulating prostaglandin production (PGI2 and PGE2), prior exposure of endothelial cells to lower doses of H2O2 (less than 0.1 microM) at high oxygen tension inhibited the subsequent stimulation of prostaglandin production by ATP, A23187, and H2O2 itself. We conclude that H2O2 has substantial effects on endothelial physiology at doses up to 3,000-fold lower than those which induce cytotoxicity.

Animals↗
PubMed1984

Effects of neutrophil elastase and other proteases on porcine aortic endothelial prostaglandin I2 production, adenine nucleotide release, and responses to vasoactive agents.

The effects of neutrophil elastase on endothelial prostacyclin (PGI2) production, nucleotide release, and responsiveness to vasoactive agents were compared with the effects of cathepsin G (the other major neutral protease of neutrophils), pancreatic elastase, trypsin, chymotrypsin, and thrombin. PGI2 production by pig aortic endothelial cells cultured on microcarrier beads and perfused in columns was stimulated in a dose-dependent manner by trypsin, chymotrypsin, and cathepsin G (1-100 micrograms/ml for 3 min). Thrombin, while active at low concentrations (0.1-10 National Institutes of Health U/ml), induced smaller responses. Neutrophil and pancreatic elastase had little or no effect on PGI2 production. Dose-dependent, selective release of adenine nucleotides was induced by neutrophil elastase (3-30 micrograms/ml). The other proteases were much less active; for example, trypsin (100 micrograms/ml) induced a response only approximately 5% as great as did 30 micrograms/ml neutrophil elastase. After exposure to 30 micrograms/ml neutrophil elastase, cells did not exhibit the characteristic burst of PGI2 production in response to extracellular ATP; responsiveness gradually returned after 40-120 min. This effect was not seen with the other proteases. Elastase partly inhibited responses to bradykinin and had no effect on PGI2 production that was stimulated by ionophore A23187. There was no evidence of cytotoxicity, as measured by release of lactate dehydrogenase. Neutrophil degranulation can generate concentrations of elastase and cathepsin G comparable with those tested in the present study, and the effects of these enzymes on endothelial function lead us to suggest that they may play a role in vasoregulation and vascular pathology.

Adenine Nucleotides↗
PubMed1983

Stereoselectivity of ectonucleotidases on vascular endothelial cells.

We have investigated the stereoselectivity of ectonucleotidases (nucleoside triphosphatase, EC 3.6.1.15; nucleoside diphosphatase, EC 3.6.1.6; 5'-nucleotidase, EC 3.1.3.5) on pig aortic endothelial cells using two classes of nucleotide analogue. In experiments with nucleotide enantiomers in which the natural D-ribofuranosyl moiety is replaced by an L-ribofuranosyl moiety, the rate of catabolism of 100 microM-L-ATP was one-fifth that of D-ATP, the rate of catabolism of 100 microM-L-ADP was one-fifteenth that of D-ADP and there was no detectable catabolism of 100 microM-L-AMP. Each of the L-enantiomers inhibited, apparently competitively, the catabolism of the corresponding D-enantiomer; Ki values were approx. 0.6 mM, 1.0 mM and 3.9 mM for L-ATP, L-ADP and L-AMP respectively. Experiments with adenosine 5'-[beta, gamma-imido]triphosphate and with D- and L-enantiomers of adenosine 5'-[beta, gamma-methylene]triphosphate revealed modest ectopyrophosphatase activity, undetectable in experiments with natural nucleotides, which was also stereoselective. Use of phosphorothioate nucleotide analogues demonstrated that ATP catabolism was virtually stereospecific with respect to the geometry of the thiol group substituted on the beta-phosphate: the Rp isomer was degraded, whereas there was little or no breakdown of the Sp isomer. ADP catabolism was also stereospecific with respect to the geometry of the thiol group substituted on the alpha-phosphate: the Sp isomer but not the Rp isomer was degraded. The geometry of thiol-group substitution on the alpha-phosphate had no effect on ATP catabolism to ADP. There was no detectable catabolism of analogues with thiol-group substitution on the terminal phosphate. Each of the phosphorothioate analogues that was catabolized broke down at a rate similar to that of the natural nucleotide from which it was derived. These results demonstrate that the ectonucleotidases on pig aortic endothelial cells exhibit a high degree of stereoselectivity, characteristic for each enzyme, both with respect to the ribofuranosyl moiety and to the phosphate side chain.

5'-Nucleotidase↗