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Biomedical subjects

I L Buxton

Publications and source records attributed to I L Buxton.

At least 19 recordsLinked to original sources
PubMed2001

NO-induced relaxation of labouring and non-labouring human myometrium is not mediated by cyclic GMP.

1. In myometrial strips from near-term non-labouring human uterus, addition of oxytocin (OT) evoked dose-dependent (10 - 3000 nM) phasic contractions that were antagonized by atosiban (1 microM) and relaxed by addition of the nitric oxide donor S-nitroso L-cysteine (Cys-NO). In near-term labouring myometrium, however, addition of OT was ineffective at raising additional tone. 2. In both labouring and non-labouring tissue, Cys-NO mediated relaxation of spontaneous or OT-induced contractions (IC(50)=1 microM) was unaffected by prior addition of the guanylyl cyclase (GC) inhibitors ODQ (1H-[1,2,4]oxadiazolo[4,3,-alpha]quinoxalin-1-one; 1 microM), or methylene blue (MB; 10 microM). 3. Elevation of intracellular cyclic GMP accompanying 30 microM Cys-NO addition in non-labouring tissue (7.5 fold) or in labouring tissues (2.5 fold) was completely blocked in tissues that had been pre-treated with ODQ or MB. 4. Charybdotoxin (ChTx), iberiotoxin (IbTx) and kaliotoxin (KalTx) all shifted the Cys-NO inhibition curve to the right and reduced the degree of relaxation produced by maximal Cys-NO treatment (100 microM in non-labouring tissue; in labouring tissue, KalTx prevented Cys-NO mediated relaxation in both stimulated and unstimulated tissue. 5. Addition of the NO-donor S-nitroso N-acetyl penicillamine (SNAP) produced a dose-dependent relaxation of pregnant myometrium while 3-morpholinosyndonimine (SIN-1) did not. The failure of SIN-1 to relax OT-induced contractions was not due to a failure of the donor to stimulate myometrial GC. 6. We demonstrate that despite the ability of NO to stimulate myometrial GC in pregnant uterine muscle, relaxations are independent of cyclic GMP action. Effects of K(+)-channel inhibitors suggests that NO-induced relaxation in human uterine smooth muscle may be subserved by direct or indirect activation of one or more calcium-activated K(+)-channels.

Charybdotoxin↗
PubMed2001

Evidence supporting the Nucleotide Axis Hypothesis: ATP release and metabolism by coronary endothelium.

The Nucleotide Axis Hypothesis, defined and supported herein, proposes that ATP stimulates the release of vasoactive mediators from endothelium, including ATP itself. Here, we show rapid endothelium-dependent, agonist-stimulated ATP elaboration in coronary vessels of guinea pigs. Measurement of extracellular ADP metabolism in intact vessels results in the time- and substrate-dependent formation of ATP in the coronary perfusate in amounts greater than can be accounted for by release from endothelium alone. ATP formation by endothelial cells is saturable (K(M) = 38.5 micromol/l, where K(M) is substrate concentration at which rate is half-maximal.) and trypsin-sensitive, membranes from [gamma-(32)P]ATP-labeled cells support ADP-dependent transphosphorylation by a 20-kDa protein, Western blots reveal the presence of a nucleoside diphosphate kinase (NDPK) of approximately 20 kDa in endothelial membranes, and analysis of NDPK antibody binding by flow cytometry is consistent with the presence of an ecto-NDPK on cardiac endothelial cells. Sequencing of the endothelial cell ecto-NDPK reveals a predicted amino acid sequence with 85% identity to human Nm23-H1 and consistent with a protein whose properties may confer membrane association as well as sites of regulation of activity. Our data underscore the potential importance of a nucleotide axis in cardiac blood vessels.

Adenosine Triphosphate↗
PubMed2001

Hypertension as a complication of topical testosterone therapy.

OBJECTIVE: To report a patent developing hypertension following the use of testosterone gel. CASE SUMMARY: A 65-year-old white man who was placed on topical testosterone therapy to increase libido developed hypertension and increased hematocrit simultaneously. After discontinuation of the testosterone, the hematocrit returned to mid-normal reference range and blood pressure returned to normal; he had been normotensive prior to topical testosterone use. DISCUSSION: The popularity of hormone replacement for women for the maintenance of appearance and lifestyle and the prevention of complications such as osteoporosis has been established over several decades, and the complications and risks and benefits of this therapy have been well described. More recently, and particularly with the availability of topical preparations of testosterone, hormone replacement for men is becoming popular with both older male patients and their physicians. We describe a case of hypertension as a complication of the application of topical testosterone and link this adverse effect to the known anabolic effects of testosterone. CONCLUSIONS: Clinicians should be cautious in prescribing topical preparations of testosterone for the proper administration by the patient and alert to the development of hypertension in this population at increased risk for cardiovascular events due to their age.

Administration, Topical↗
PubMed2000

Role of nucleotides and nucleosides in the regulation of cardiac blood flow.

The regulation of blood flow in the heart on a moment-to-moment basis is essential to meet changes in the oxygen demands of cardiac muscle. The signals that subserve this regulation are not all firmly established. Although the formation and release of adenosine by cardiac muscle during periods of hypoxia or regional ischemia in the heart are well known to produce regional vasodilation and salvage of at-risk myocardium, these extracellular actions of adenosine are believed to occur abluminally and thus do not explain the origin or predict the potent actions of intravascular adenosine. The notion that purines such as adenosine and adenosine 5'-triphosphate (ATP) might be available to act in the lumen of the blood vessel has been proposed by the authors and others to help explain the regulation of blood flow in the heart in nonpathologic states. This article details the background and current understanding of the vascular actions of adenosine and ATP, defines the Nucleotide Axis Hypothesis, and reviews clinical studies in which its likely importance in the maintenance of blood flow in the heart has been investigated.

Coronary Circulation↗
PubMed2000

Regulation of uterine contraction: mechanisms in preterm labor.

Preterm labor (PTL) is defined as uterine irritability accompanied by cervical dilation and/or effecement that occurs before 37 weeks gestation. In most cases, PTL becomes preterm delivery (PTD), accounting for 8% to 10% of births in the United States. Fetuses born before 37 weeks' gestation are at risk for a multitude of health and developmental problems. Most perinatal morbidity and mortality in the United States are caused by PTL. It is a costly problem, in both monetary and human terms. Although some risk factors have been identified, they by no means identify, in advance, every case of PTL and PTD. Despite the understandable emphasis on attempts to find and test risk factors that predict PTL, the ultimate benefit--preventing PTD--will come only from an understanding of the physiologic mechanisms of parturition and how to halt those processes when they occur too early. This article reviews current approaches to preventing PTD, describes the biology of myometrial contraction, and discusses recent progress from several laboratories including the authors' that may shed light on approaches to inhibit uterine contractility in the setting of PTL.

Female↗
PubMed1998

Nitric oxide regulation of monkey myometrial contractility.

1. We evaluated the effect of the nitric oxide (NO) donor CysNO (S-nitroso-L-cysteine) and endogenous NO upon spontaneous contractility in non-pregnant cynomolgus monkeys. We also assessed the role of intracellular guanosine 3',5'-cyclic monophosphate ([cyclic GMP]i) as a second messenger for NO in monkey uterine smooth muscle. 2. CysNO reduced spontaneous contractility by 84% (P < 0.05) at maximal concentrations, and significantly elevated [cyclic GMP]i (P < 0.05). However, increases in [cyclic GMP]i were not required for CysNO-induced relaxations; CysNO inhibited contractile activity despite the complete inhibition of guanylyl cyclase by methylene blue or LY83,583. 3. Analogues of cyclic GMP had no significant effect upon spontaneous contractile activity. L-arginine produced a 62% reduction in spontaneous activity (P < 0.05) while D-arginine had no effect. The competitive nitric oxide synthase (NOS) inhibitor N(omega)-nitro-L-arginine (L-NOARG) not only blocked L-arginine-induced relaxations, but also significantly increased spontaneous contractile activity when added alone (P < 0.05); the inactive D-enantiomer of NOARG had no such effect. 4. While both endogenous NO and the NO donor CysNO relax monkey myometrium, this effect is not causally related to CysNO-induced elevations in [cyclic GMP]i. The failure of cyclic GMP analogues to alter monkey uterine smooth muscle tension also argues against a role for [cyclic GMP]i in the regulation of uterine contractility. Not only do these findings argue for the existence of a functionally-relevant NOS in the monkey uterus, but increases in contractile activity seen in the presence of NOS inhibitors suggest a role for NO in the moment-to-moment regulation of contractile activity in this organ.

Animals↗
PubMed1998

Nitric oxide relaxes human myometrium by a cGMP-independent mechanism.

The role of intracellular guanosine 3',5'-cyclic monophosphate concentration ([cGMP]i) in nitric oxide (NO)-mediated relaxations in the uterus has become controversial. We found the NO donor S-nitroso-L-cysteine (CysNO) to potently (IC50 = 30 nM) inhibit spontaneous contractions in the nonpregnant human myometrium. CysNO treatment increased [cGMP]i significantly (P < 0.001), and this increase was blocked by the guanylyl cyclase inhibitors methylene blue (10 microM) or LY-83583 (1 microM); however, pretreatment with these guanylyl cyclase inhibitors failed to block CysNO-mediated relaxations. Intracellular cAMP concentrations were not altered by treatment of tissues with 10 microM CysNO. Incubation with the cGMP analogs 8-bromo-cGMP or beta-phenyl-1,N2-etheno-cGMP did not significantly affect spontaneous contractility. Pretreatment of tissues with charybdotoxin [a calcium-dependent potassium channel (BK) blocker] completely reversed CysNO-induced relaxations. We conclude that NO is a potent inhibitor of spontaneous contractile activity in the nonpregnant human uterus and that, although guanylyl cyclase and BK activities are increased by NO, increases in [cGMP]i are not required for NO-induced relaxations in this tissue.

Aminoquinolines↗
PubMed1997

Competition of adenine nucleotides for a 1,3-[3H]-dipropyl-8-cyclopentylxanthine binding site in rat vas deferens.

1. The binding of 1,3-[3H]-dipropyl-8-cyclopentylxanthine ([3H]-DPCPX), a specific adenosine A1 receptor antagonist, was examined in rat vas deferens membrane preparations using radioligand binding techniques. 2. 1,3-[3H]-Dipropyl-8-cyclopentylxanthine bound to these preparations with a KD of 1.07 +/- 0.14 nmol/L (n = 6). The density of [3H]-DPCPX binding sites was 133.38 +/- 5.57 fmol/mg protein. 3. Computer analysis indicated that nucleosides competed for [3H]-DPCPX binding at two distinct sites. The rank order of potency at the higher affinity site corresponded to R-phenylisopropyladenosine (R-PIA) > or = 2-chloroadenosine (2-CIADO) > or = cyclopentyladenosine (CPA) > or = N-ethylcarboxamidoadenosine (NECA) > s-phenylisopropyladenosine (s-PIA). Ki values were in the low nmol/L range. The rank order of nucleoside potency at the lower affinity site corresponded to R-PIA > or = CPA > or = NECA > or = 2-CIADO > S-PIA. Ki values were in the low mumol/L range. 4. Nucleotides competed for [3H]-DPCPX binding at a single site only. The rank order of potency at this site corresponded to alpha, beta-methylene ATP > or = beta, gamma-methylene ATP > or = ATP. Ki values were in the high mumol/L range. The site seemed to correspond with one of the two binding sites predicted by nucleoside competition binding. 5. The ATP-regenerating compound myokinase did not significantly change the competition curve for ATP, indicating that the competition for [3H]-DPCPX binding observed in the presence of ATP was due to an effect of ATP per se and not to an action of a degradation product. 6. The results demonstrate that in rat vasa deferentia there exist two distinct binding sites for [3H]-DPCPX. One of these sites binds only nucleosides and may represent an adenosine A1 receptor, as usually defined. The other site binds both nucleosides and nucleotides and may represent an atypical adenosine A1 receptor, an atypical P2 or a P3 purinoceptor.

2-Chloroadenosine↗
PubMed1996

HHMI awards.

Explore the source record for details and available documents.

Academies and Institutes↗
PubMed1996

Evidence for a discrete UTP receptor in cardiac endothelial cells.

1. We have examined the effects of various purine and pyrimidine nucleotides upon cells cultured from guinea-pig cardiac endothelium (CEC), and find the P2Y-agonist 2-methylthioadenosine triphosphate (2MeSATP) to be a potent (EC50 = 85 +/- 10.2 nM) stimulator of increase in intracellular calcium concentrations, while uridine 5'-triphosphate (UTP) and adenosine 5'-triphosphate (ATP) are less potent but equipotent with one another (EC50s = 2.1 +/- 0.3 and 1.8 +/- 0.2 microM, respectively). 2. While the P2Y receptor exhibited rapid homologous desensitization, this had no effect upon subsequent responsiveness of CEC to either ATP or UTP. Effects of maximal concentrations of ATP and UTP were not only additive, but did not cross-desensitize. Responses to UTP (but not to ATP or 2MeSATP) were blocked by treatment with pertussis toxin (PTX); all three nucleotides appeared to liberate calcium from an intracellular pool. 3. Suramin (30 microM) significantly (P < 0.05) increased the EC50 for ATP-dependent increases in intracellular calcium (5.3 +/- 2.2 microM vs. 2.0 +/- 0.9 microM in the absence of suramin), while it completely blocked the response to 2MeSATP. Suramin had no effect upon responses to UTP at concentrations of 100 microM. 4. We conclude that in addition to the P2Y and P2U subtypes of the ATP receptor, an additional receptor responsive to UTP but exhibiting no affinity for purine nucleotides is present in CEC; this "pyrimidine receptor' liberates intracellular calcium via a G-protein, and may partly mediate the contractile response to UTP in the coronary vasculature.

Adenosine Triphosphate↗
PubMed1996

Cyclic GMP-independent effects of nitric oxide on guinea-pig uterine contractility.

1. The role of nitric oxide (NO) in the regulation of uterine contractility has yet to be clearly defined. We evaluated the effect of NO (in the form of S-nitroso-cysteine, CysNO) upon uterine contractility and guanosine 3',5'-cyclic monophosphate (cyclic GMP) accumulation in pregnant and nonpregnant guinea-pig myometrium. 2. While CysNO had no effect upon spontaneous contractile activity in either pregnant or nonpregnant uterine tissues, addition of CysNO resulted in an immediate and reversible relaxation of oxytocin- or acetylcholine (ACh)-evoked contractions. 3. Relaxation of agonist-evoked contractions in response to CysNO was associated with significant elevations in intracellular cyclic GMP concentrations ([cyclic GMP]i). 4. Elevations in [cyclic GMP]i were not required for relaxation, as inhibition of guanylyl cyclase by methylene blue prevented [cyclic GMP]i accumulation while having no effect upon the ability of CysNO to relax agonist-evoked contractions. 5. Addition of the cyclic GMP-analogues, 8-Br-cyclic GMP and PET-cyclic GMP, only at high concentrations, produced partial relaxation of agonist-contracted tissues, suggesting the possibility that cyclic GMP may be sufficient but not necessary for myometrial relaxation. 6. Our studies not only provide evidence for a functional role for NO-modulation of agonist-evoked contractions in the pregnant and nonpregnant guinea-pig uterus, but also that these occur by a mechanism which is not dependent upon guanylyl cyclase activity.

Animals↗
PubMed1995

Detection of nitric oxide release from canine enteric neurons.

Previous pharmacological and immunohistochemical studies have suggested that nitric oxide (NO) is a mediator of enteric neuromuscular transmission in the canine proximal colon. The present study demonstrates the release of nitric oxide (NO) and/or its oxidation products (collectively termed NOx) following stimulation of intrinsic neurons with the nicotinic agonist 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP). Strips of muscularis externa, including the ganglionated myenteric and submucosal plexuses were suspended under tension in modified Krebs solution. DMPP (1-100 microM) produced concentration-dependent inhibition of circular muscle contractile activity and this effect was antagonized by L-nitroarginine methyl ester or L-nitroarginine (100 microM). Tetrodotoxin (TTX, 1 microM) significantly reduced mechanical responses to DMPP (10 microM) but had no effect on responses to high concentrations of DMPP (100 microM). Aliquots of the bathing medium were assayed for NOx after regeneration of NO from NO2- and/or NO3-. NO was measured as chemiluminescence produced by reaction with ozone. Detection was linear over the range 6-25,000 pmol of added NO2- or NO3-. In the absence of drugs, a basal release of 1113.5 +/- 100.4 pmol NOx/g tissue (n = 27) was detected after a 30-min incubation period. DMPP (1-100 microM) stimulated a concentration-dependent increase in NOx to 5099.9 +/- 430 pmol/g per 30 min (n = 17) at 100 microM. NOx release was inhibited by an inhibitor of nitric oxide synthase activity (N(G)-monomethyl-L-arginine) or by reduction in extracellular Ca2+ concentration. DMPP-stimulated NOx accumulation was significantly reduced, but not abolished by TTX (1 microM). These results provide further evidence that NO is released following stimulation of intrinsic neurons in canine proximal colon and support previous suggestions that nicotinic agonists may directly stimulate terminals of NO-releasing neurons.

Animals↗