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Biomedical subjects

M B Murphy

Publications and source records attributed to M B Murphy.

At least 37 records · Page 2Linked to original sources
PubMed1999

Biochemical characterization of a Dictyostelium myosin II heavy-chain phosphatase that promotes filament assembly.

In Dictyostelium cells, myosin II is found as cytosolic nonassembled monomers and cytoskeletal bipolar filaments. It is thought that the phosphorylation state of three threonine residues in the tail of myosin II heavy chain regulates the molecular motor's assembly state and localization. Phosphorylation of the myosin heavy chain at threonine residues 1823, 1833 and 2029 is responsible for maintaining myosin in the nonassembled state, and subsequent dephosphorylation of these residues is a prerequisite for assembly into the cytoskeleton. We report here the characterization of myosin heavy-chain phosphatase activities in Dictyostelium utilizing myosin II phosphorylated by myosin heavy-chain kinase A as a substrate. One of the myosin heavy-chain phosphatase activities was identified as protein phosphatase 2A and the purified holoenzyme was composed of a 37-kDa catalytic subunit, a 65-kDa A subunit and a 55-kDa B subunit. The protein phosphatase 2A holoenzyme displays two orders of magnitude higher activity towards myosin phosphorylated on the heavy chains than it does towards myosin phosphorylated on the regulatory light chains, consistent with a role in the control of filament assembly. The purified myosin heavy-chain phosphatase activity promotes bipolar filament assembly in vitro via dephosphorylation of the myosin heavy chain. This system should provide a valuable model for studying the regulation and localization of protein phosphatase 2A in the context of cytoskeletal reorganization.

Actin Cytoskeleton↗
PubMed1999

Opioid antagonist modulation of ischaemia-induced ventricular arrhythmias: a peripheral mechanism.

Ventricular arrhythmias are an important cause of death after myocardial ischaemia. Animal studies have generated conflicting data on the potentiating or attenuating effects of opioid agonists and antagonists on cardiac rhythm during acute myocardial ischaemia and coronary artery reperfusion. Whether these effects of opioid antagonists are mediated by central or peripheral nervous system mechanisms remains unclear. We examined (a) the effects of peripheral opioid receptor blockade on ischaemia-induced arrhythmia by using methylnaltrexone (MNTX), a novel quaternary derivative of naltrexone (NTX) that does not cross the blood-brain barrier, and (b) whether MNTX would modulate morphine effects during acute coronary artery ligation and reperfusion in the rabbit. The incidence and severity of cardiac arrhythmias were assessed during 40 min of coronary artery occlusion and reperfusion and summarised in an arrhythmia score (AS). MNTX reduced the incidence of ventricular fibrillation (VF) and arrhythmia score during coronary artery occlusion when compared with vehicle (p < 0.05). Naltrexone reduced the incidence of VF (p < 0.05). Although morphine alone had no significant effects, its coadministration blunted the antiarrhythmic properties of MNTX. The data suggest that blockade of opiate receptors outside the central nervous system may protect against ischaemia-induced arrhythmias.

Animals↗
PubMed1999

Short- and long-term outcome of total parathyroidectomy with immediate autografting versus subtotal parathyroidectomy in patients with end-stage renal disease.

A retrospective study was performed in 36 patients with end-stage renal disease (ESRD) comparing total parathyroidectomy followed by immediate autografting into the forearm (total PTX + IA) with parathyroidectomy (subtotal PTX) over a five-year period. Twenty-eight patients underwent subtotal PTX and 8 had total PTX + IA. The two surgical methods were evaluated with respect to preoperative severity of hyperparathyroidism, perioperative morbidity, and the incidence of recurrent hyperparathyroidism. Eleven patients in total (30.6%) developed recurrent hyperparathyroidism; 2/8 (25%) in the total PTX + IA group compared to 9/28 (32.1%) in the subtotal PTX group (p = 0.699). The median time to recurrence was longer in the total PTX + IA group (39 vs. 16 months), and the median long-term postoperative PTH value was lower (81 vs. 199 ng/l), but these differences did not reach statistical significance. In conclusion, the incidence of recurrent hyperparathyroidism is high regardless of surgical modality. However, total PTX + IA may produce more favorable results with respect to median postoperative PTH level and time to recurrence.

Alkaline Phosphatase↗
PubMed1997

Mapping of the novel protein kinase catalytic domain of Dictyostelium myosin II heavy chain kinase A.

Myosin heavy chain kinase A (MHCK A) in Dictyostelium was identified as a biochemical activity that phosphorylates threonine residues in the myosin II tail domain and regulates myosin filament assembly. The catalytic domain of MHCK A has now been mapped through the functional characterization of a series of MHCK A truncation mutants expressed in Escherichia coli. A recombinant protein comprising the central nonrepetitive domain of MHCK A (residues 552-841) was isolated in a soluble form and shown to phosphorylate Dictyostelium myosin II, myelin basic protein, and a synthetic peptide substrate. The functionally mapped catalytic domain of MHCK A shows no detectable sequence similarity to known classes of eukaryotic protein kinases but shares substantial sequence similarity with a transcribed Caenorhabditis elegans gene and with the mammalian elongation factor-2 kinase (calcium/calmodulin-dependent protein kinase III). We suggest that MHCK A represents the prototype for a novel, widely occurring protein kinase family.

Amino Acid Sequence↗
PubMed1997

Factors confounding assessment of ambulatory blood pressure monitors, studied during formal evaluation of the Tycos Quiet-Trak.

The credibility of studies assessing ambulatory blood pressure monitoring (ABPM) devices will be enhanced by minimizing opportunities to manipulate the outcome through biased selection of patient volunteers, and by insuring that participants are representative of the target patient population. While subjecting the Welch Allyn Quiet-Trak to the British Hypertension Society (BHS) protocol, we examined the extent to which certain candidate characteristics might influence device assessment. The Quiet-Trak achieved an A grade overall. During its field testing, daytime physical activity measured with a wrist-mounted, piezoelectric accelerometer, influenced significantly the rate of measurement rejection. The tertiles of subjects with highest and lowest levels of daytime physical activity (64 +/- 16 and 35 +/- 10 activity units; P < .001) exhibited significantly different measurement rejection rates (10 +/- 3 and 3 +/- 2 daytime rejects; P < .001)). Most rejected readings occurred during episodes of high physical activity. During static evaluation (Phase V), the level of systolic BP influenced the accuracy of diastolic BP estimation; above 190 mm Hg systolic BP, estimates of diastolic BP differed significantly from manual measurements. Subjects' age and arm circumference influenced neither field nor static evaluation. Retrospective comparison of the study subjects with 120 consecutive ABPM clinic attenders revealed (1) that participants in field testing were younger, had lower BP but were equally active compared with clinic patients; and (2) that Phase V participants exhibited higher systolic (155 +/- 42 v 135 +/- 15 mm Hg; P < .001) but similar diastolic BP levels compared to controls. The reliability of ABPM validation protocols would be enhanced by: (a) incorporation of objective measurement of physical activity during field testing, demonstrating normal levels of activity; (b) requiring that field testing be conducted in representative patients rather than normal volunteers; and (c) a greater focus in both static and field testing on those levels of blood pressure that are relevant in clinical practice.

Blood Pressure Monitoring, Ambulatory↗
PubMed1997

Opioid-induced delay in gastric emptying: a peripheral mechanism in humans.

BACKGROUND: Opioids delay gastric emptying, which in turn may increase the risk of vomiting and pulmonary aspiration. Naloxone reverses this opiate action on gastric emptying, but it is not known whether this effect in humans is mediated by central or peripheral opiate antagonism. The importance of peripheral opioid receptor antagonism in modulating opioid-induced delay in gastric emptying was evaluated using methylnaltrexone, a quaternary derivative of the opiate antagonist naltrexone, which does not cross the blood-brain barrier. METHODS: In a randomized, double-blind, crossover placebo-controlled study, 11 healthy volunteers were given either placebo (saline), 0.09 mg/kg morphine, or 0.09 mg/kg morphine plus 0.3 mg/kg methylnaltrexone on three separate occasions before ingesting 500 ml deionized water. The rate of gastric emptying was measured by two methods: a noninvasive epigastric bioimpedance technique and the acetaminophen absorption test. RESULTS: The epigastric bioimpedance technique was sufficiently sensitive to detect opioid-induced changes in the rate of gastric emptying. The mean +/- SD time taken for the gastric volume to decrease to 50% (t0.5) after placebo was 5.5 +/- 2.1 min. Morphine prolonged gastric emptying to (t0.5) of 21 +/- 9.0 min (P < 0.03). Methylnaltrexone given concomitantly with morphine reversed the morphine-induced delay in gastric emptying to a t0.5 of 7.4 +/- 3.0 (P < 0.04). Maximum concentrations and area under the concentration curve from 0 to 90 min of serum acetaminophen concentrations after morphine were significantly different from placebo and morphine administered concomitantly with methylnaltrexone (P < 0.05). No difference in maximum concentration or area under the concentration curve from 0 to 90 min was noted between placebo and methylnaltrexone coadministered with morphine. CONCLUSIONS: The attenuation of morphine-induced delay in gastric emptying by methylnaltrexone suggests that the opioid effect is mediated outside the central nervous system. Methylnaltrexone may have the potential to decrease the side effects of opioid medications, which are mediated peripherally, while maintaining the central analgesia effect of the opioid.

Acetaminophen↗
PubMed1997

A comparison of the effects of tramadol and morphine on gastric emptying in man.

In a previous study using an electrical bioimpedance technique and the paracetamol absorption test, we demonstrated that 0.09 mg.kg-1 of morphine delayed gastric emptying in healthy human volunteers. The aim of this study was to investigate whether analgesic doses of tramadol would cause a delay in gastric emptying similar to conventional opioids. Using the same volunteers and techniques as in our previous study, placebo or tramadol (1 mg.kg-1) was given in a randomised, double-blinded, cross-over placebo-controlled study. Gastric emptying was measured concurrently by a noninvasive epigastric bioimpedance technique and by the paracetamol absorption test. After the ingestion of 500 ml of deionised water plus paracetamol 1.5 g, the mean (SEM) time taken for gastric volume to decrease to 50% (t0.5) was recorded. No difference in gastric emptying rates (t0.5) between placebo, 7.7 (1 min), and tramadol, 9.5 (2 min), was noted. In our previous study, morphine prolonged t0.5 to 21 (3) min (p < 0.03). The maximum concentration and area under the curve of serum paracetamol concentrations following morphine were significantly different from placebo (p < 0.05) and tramadol (p < 0.05). We conclude that tramadol at a dose of 1 mg.kg-1 does not delay gastric emptying in humans.

Absorption↗
PubMed1995

Differential effects of chronic oral antihypertensive therapies on systemic arterial circulation and ventricular energetics in African-American patients.

BACKGROUND: A comprehensive evaluation of arterial load characteristics and left ventricular energetics in systemic hypertension has been limited by the need for invasive techniques to access instantaneous aortic pressure and flow. As a consequence of this methodological limitation, no data exist on the effects of long-term antihypertensive therapy on global arterial impedance properties and indexes of myocardial oxygen consumption (MVO2). Using recently validated noninvasive techniques, we compared in hypertensive patients the effects of chronic oral treatment with ramipril, nifedipine, and atenolol on arterial impedance and mechanical power dissipation as well as indexes of MVO2. METHODS AND RESULTS: Sixteen African-American subjects with systemic hypertension were studied with a randomized, double-blind, crossover protocol. Instantaneous central aortic pressure and flow, from which arterial load characteristics can be derived, were estimated from calibrated subclavian pulse tracings (SPTs) and continuous-wave aortic Doppler velocity in conjunction with two-dimensional (2D) echocardiographic measurements of the aortic annulus, respectively. To derive ventricular wall stress and indexes of MVO2, left ventricular short- (M-mode) and long-axis (2D echo) images were acquired simultaneously with SPTs. Data were collected at the end of a 2-week washout period (predrug control) and after 6 weeks of treatment with each agent. Although all three agents reduced diastolic blood pressure to the same extent, different effects on mean and systolic pressures and vascular impedance properties were noted. Nifedipine reduced total peripheral resistance (TPR; 1744 +/- 398 versus 1290 +/- 215 dyne-s/cm5) and increased arterial compliance (ACL; 1.234 +/- 0.253 versus 1.776 +/- 0.415 mL/mm Hg). This improvement in arterial compliance was not entirely accounted for by the reduction in distending pressure. Ramipril also decreased TPR (1740 +/- 292 versus 1437 +/- 290 dyne-s/cm5) and increased ACL (1.214 +/- 0.190 versus 1.569 +/- 0.424 mL/mm Hg), but with this agent, the change in arterial compliance was explained solely on the basis of a reduction in distending pressure. Atenolol, in contrast, did not affect either TPR or ACL. In agreement with the compliance results, nifedipine and ramipril significantly lowered the first two harmonics of the impedance spectrum, but atenolol did not. None of these agents resulted in a significant change in characteristic impedance or in the relative amplitude of the reflected pressure wave. Total vascular mechanical power and percent of oscillatory power remained unaltered with all antihypertensive treatments. Only ramipril and nifedipine reduced the integral of both meridional and circumferential systolic wall stresses, indicating that MVO2 per beat was reduced with these agents. Stress-time index, a measure of MVO2 per unit time, decreased significantly with ramipril but not with nifedipine because of an increase in heart rate noted in 10 of 16 patients (mean increase, 10 beats per minute). Thus, a reduction in MVO2 coupled with unchanged total vascular mechanical power suggests improved efficiency of ventriculoarterial coupling with ramipril and with nifedipine in the subset of patients in whom heart rate remained unchanged. In contrast, there was no evidence of a reduction in wall stress, stress integral, or stress-time index with atenolol. CONCLUSIONS: The noninvasive methodology used in this study constitutes a new tool for serial and simultaneous evaluation of arterial hemodynamics and left ventricular energetics in systemic hypertension. In this study, we demonstrate the differential effects of chronic antihypertensive therapies on systemic arterial circulation and indexes of MVO2 in African-American subjects. Consideration of drug-induced differential responses of arterial load and indexes of MVO2 with each drug may provide a more physiological approach to the treatment of systemic hypertension in indivi

Administration, Oral↗
PubMed1995

Prostacyclin and thromboxane levels in pleural space fluid during cardiopulmonary bypass.

Prostaglandins exhibit a variety of cardiovascular actions that may affect the hemodynamic recovery of the ischemic myocardium after cardiopulmonary bypass. We have observed a decrease in the mean arterial pressure on autotransfusion of the accumulated pleural cavity fluid during operation. One aim of this study was to determine the concentrations of prostacyclin and thromboxane A2 in the pleural cavity fluid by measuring their stable metabolites, 6-keto-PGF1 alpha and thromboxane B2, respectively, in 8 consecutive patients undergoing myocardial revascularization, and to compare them with the arterial levels. A second aim was to quantify the hemodynamic effect of the pleural cavity fluid during operation. The concentration of 6-keto-PGF1 alpha in the pleural cavity fluid was significantly higher than the arterial concentration (mean, 21.6 +/- 18.2 ng/mL; p < 0.01). The concentration of thromboxane B2 was also raised compared with the arterial concentration (mean, 3.62 +/- 5.96 ng/mL; p < 0.2). The percentage fall in the mean arterial pressure was 29.7% +/- 8.86% (p < 0.02), which was transient and lasted 1 to 3.5 minutes. The hemoglobin concentration, potassium level, and pH were also measured. This study shows that the pleural cavity fluid during cardiac operations contains significant amounts of endogenous prostacyclin. Considering the potential benefit of prostacyclin on the recovering myocardium, we believe that this fluid should be transfused as a volume replacement, keeping in mind the transient phase of hemodynamic instability.

6-Ketoprostaglandin F1 alpha↗
PubMed1994

Ethnic differences in circadian hemodynamic profile.

Since the introduction of 24-h ambulatory blood pressure monitoring (ABPM), some studies, although not all, have suggested the presence of a blunted nocturnal blood pressure decline in black versus white subjects, a difference that may help explain the higher incidence of target organ damage in blacks. To better define ethnic differences in diurnal hemodynamic profiles, we studied ABPM recordings from 275 black (55.6% women) and 246 white (43.1% women) previously untreated subjects, with a similar age distribution (from 20 to 79 years) and a wide range of systolic (100-230 mm Hg) and diastolic (50-130 mm Hg) blood pressures. Average clinic systolic (diastolic) blood pressures were higher in black v white men by 10.2 (7.3) mm Hg; P = .04 (P = .004), with a similar trend in women (P = NS). On ABPM, blacks had higher average values, a difference that was greater during sleep (9.4 mm Hg for systolic blood pressure) than while awake (4.7 mm Hg; P = .003). Average diurnal change in systolic blood pressure (awake minus sleep values) was 13.1 +/- 0.7 v 18.0 +/- 0.6 mm Hg for blacks v whites (P < .001). There was a strong negative correlation between baseline (ie, sleep) blood pressure and the diurnal change (r = -0.58; P < .001), but at each given level, blacks had a lower daytime increment/nocturnal fall (P = .02). Results for diastolic blood pressure and heart rate were similar. The data suggest that the smaller diurnal change in blacks may be related in part to their higher blood pressure levels, but that there is an additional, independent effect of race. This results in a greater 24-h blood pressure load in blacks than whites for each given clinic (daytime) value, and may help explain differences in target organ damage. Future studies investigating the effects of blood pressure on target organs in different populations should consider diurnal profiles in addition to clinic blood pressure.

Adult↗
PubMed1994

The dual-specificity CLK kinase induces neuronal differentiation of PC12 cells.

CLK is a dual-specificity protein kinase capable of phosphorylating serine, threonine, and tyrosine residues. We have investigated the action of CLK by establishing stable PC12 cell lines capable of inducibly expressing CLK. Expression of CLK in stably transfected PC12 cells mimicked a number of nerve growth factor (NGF)-dependent events, including the morphological differentiation of these cells and the elaboration of neurites. Moreover, CLK expression enhanced the rate of NGF-mediated neurite outgrowth of these cells, indicating that CLK expression and NGF treatment activate similar signal transduction pathways. CLK expression, unlike NGF, was not able to promote PC12 cell survival in serum-free media, demonstrating that CLK only partially recapitulated the actions of NGF on these cells and that the biochemical pathways necessary for morphological differentiation can be stimulated without also stimulating those necessary for survival. Induction of CLK expression also resulted in the selective activation of protein kinases that are components of growth factor-stimulated signal transduction cascades, including ERK1, ERK2, pp90RSK, and S6PKII. Induction of CLK expression, however, did not stimulate pp70S6K or Fos kinase, two NGF-sensitive protein kinases. These data indicate that CLK action mediates the morphological differentiation of these cells through its capacity to independently stimulate signal transduction pathways normally employed by NGF.

Animals↗
PubMed1994

The use of ambulatory blood pressure monitoring in the evaluation of racial differences in blood pressure.

Ethnic differences in 24h blood pressure profiles exist, and 24h ambulatory blood pressure monitoring shows that blacks have a higher nocturnal blood pressure associated with a small diurnal change in pressure. Studies in adolescents suggest that these diurnal differences antedate adulthood. Nocturnal blood pressure may be independently important in the pathogenesis of left ventricular hypertrophy.

Adolescent↗
PubMed1993

Identification and characterization of a nerve growth factor-stimulated mitogen-activated protein kinase activator in PC12 cells.

Nerve growth factor treatment of PC12 cells results in the rapid activation of MAP kinases. These enzymes are activated through interaction with a protein "activator." The mitogen-activated protein (MAP) kinase activator has been partially purified by ion exchange and gel filtration chromatography. The activator has an apparent molecular mass of 50-60 kDa. The MAP kinase activator is rapidly generated in response to nerve growth factor (NGF) and can be detected within 30 s of exposure, reaching maximal levels within 2 min and then declining to near basal levels by 15-20 min. The activation of MAP kinase is dependent upon the time of incubation with the activator and on activator concentration. The MAP kinase activator is itself a protein kinase that phosphorylates MAP kinases and mediates their activation. The NGF-stimulated MAP kinase activator phosphorylates MAP kinase on serine, threonine, and tyrosine residues, establishing this enzyme as dual specific kinase. The MAP kinase activator is itself a phosphoprotein whose phosphorylation on tyrosine residues is stimulated upon NGF treatment of the cells. The enzyme activity of MAP kinase activator is abolished by treatment with both the tyrosine-specific phosphatase PTP-1 and the serine/threonine-specific phosphatase PP2A. The activator is produced in response to NGF, epidermal growth factor, and fibroblast growth factor. The protein kinase inhibitor K252a selectively inhibits the ability of NGF to generate MAP kinase activator activity. These data suggest that the upstream events governing MAP kinase activation involve the regulated phosphorylation of dual specificity MAP kinase activator as an immediate consequence of receptor activation.

Animals↗
PubMed1993

Comparison of two noninvasive screening tests for renovascular hypertension.

OBJECTIVE: To compare and contrast the diagnostic accuracy rates of two newer noninvasive screening tests for renovascular hypertension, the most common curable cause of secondary hypertension. PATIENTS AND METHODS: One hundred fifty patients, thought to have a high probability of renovascular hypertension by established clinical criteria, underwent both the captopril challenge test and the renal scintigram with angiotensin-converting enzyme inhibitor, while on their usual antihypertensive regimen except angiotensin-converting enzyme inhibitors. If the result of either test was abnormal, angiography was undertaken, followed immediately by angioplasty (if a stenosis was found) or by renal vein renin determinations. Patients whose blood pressures were lower 6 to 12 weeks after a revascularization procedure (surgery or angioplasty) were diagnosed as having renovascular hypertension. RESULTS: Of the 150 patients, 100 underwent angiography, and 59 had renal artery stenosis. Of 53 patients who had surgery (n = 21) or angioplasty (n = 32), 51 had lowered blood pressures compared with before the procedure. Sensitivity and specificity of the tests were as follows: renal scintigram with angiotensin-converting enzyme inhibitor: 92% and 91% (all patients) and 92% and 80% (only patients with angiograms); captopril challenge test: 76% and 82% (all patients) and 76% and 58% (only patients with angiograms). Little difference in accuracy rates was observed in subgroup analyses in patients with chronic renal impairment, previous diuretic or beta-blocker therapy, or bilateral renal artery stenosis. CONCLUSIONS: In selected, treated patients with a high probability of renovascular hypertension, the renal scintigram with angiotensin-converting enzyme inhibitor was a more accurate noninvasive screening test than the captopril challenge test. Noninvasive screening tests for renovascular hypertension can help to identify patients who should undergo angiography and often predict success after revascularization.

Aged↗