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E Murphy

Publications and source records attributed to E Murphy.

At least 217 records · Page 12Linked to original sources
PubMed1988

Measurement of cytosolic free magnesium ion concentration by 19F NMR.

Fluorinated derivatives of the chelator o-aminophenol-N,N,O-triacetic acid (APTRA) have been developed, synthesized, and analyzed for use as 19F NMR indicators of free cytosolic magnesium concentration. Magnesium dissociation constants for the 4-fluoro, 5-fluoro, and 4-methyl-5-fluoro species were determined to be 3.1, 0.9, and 0.6 mM, respectively, on the basis of UV absorption measurements at 37 degrees C in 115 mM KCl and 20 mM NaCl, pH 7.1, buffered with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid-tris-(hydroxymethyl)aminomethane. The corresponding pK values, which reflect protonation of the nitrogen atom, were determined by 19F NMR to be 4.15, 5.45, and 5.55, respectively, so that the chelators are insensitive to pH variations near the normal physiological range. The dissociation constants of these chelators for calcium ions are lower than those for magnesium but roughly 2-3 orders of magnitude above typical basal cytosolic free calcium levels, so that calcium ions will not interfere with the determinations of magnesium levels. 19F NMR studies carried out at 339.7 MHz indicate that magnesium ions are in slow exchange with the 5-fluoro and 4-methyl-5-fluoro APTRA derivatives and in fast exchange with the 4-fluoro APTRA derivative. In contrast, calcium ions were found to be in intermediate to fast exchange with all chelators. The apparent anomaly of higher thermodynamic stability of the APTRA complexes for calcium relative to magnesium but lower kinetic stability (higher k-1 values) for the calcium complexes reflects the very different association rates for the two ions. Thus, the magnesium association rates are 3 orders of magnitude slower than those for calcium ions.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcium↗
PubMed1988

Uptake of cesium ions by human erythrocytes and perfused rat heart: a cesium-133 NMR study.

Cesium-133 NMR studies have been carried out on suspended human erythrocytes and on perfused rat hearts in media containing CsCl. The resulting spectra exhibit two sharp resonances, arising from intra- and extracellular Cs+, separated in chemical shift by 1.0-1.4 ppm. Thus, intra- and extracellular resonances are easily resolved without the addition of paramagnetic shift reagents required to resolve resonances of the other alkali metal ions. Spin-lattice relaxation times in all cases are monoexponential and significantly shorter (3-4 times) for the intracellular component. When corrections are made for the pulse repetition rate, the total intensity of the intracellular and extracellular Cs+ resonances in erythrocytes is conserved, implying total observability of the intracellular pool. The uptake of Cs+ by erythrocytes occurs at approximately one-third the reported rate for K+ and was reduced by a factor of 2 upon addition of ouabain to the sample. These results indicate that 133Cs NMR is a promising tool for studying the distribution and transport of cesium ions in biological systems and, in some cases such as uptake by cellular Na,K-ATPase, for analysis of K+ ion metabolism.

Animals↗
PubMed1988

NMR observability of ATP: preferential depletion of cytosolic ATP during ischemia in perfused rat liver.

The extent to which cellular metabolites are NMR observable is of fundamental importance in the interpretation of in vivo NMR studies. Analysis of ischemic rat liver shows that ATP resonances measured by 31P NMR decrease considerably faster than total tissue ATP measured in extracts. This discrepancy demonstrates that, in liver, ATP is not 100% observable. Furthermore, the data are consistent with the supposition that in situ mitochondrial ATP resonances are not normally observable by in vivo NMR techniques. The specificity of the NMR measurement for cytosolic ATP indicates that 31P NMR can be a valuable tool for the specific measurement of ATP in this compartment.

Adenosine Triphosphate↗
PubMed1988

Transposon Tn554 encodes three products required for transposition.

Tn554 is a high-frequency, site-specific, transposable element having integrative properties resembling those of lysogenic bacteriophages. Nucleotide sequence analysis indicates that Tn554 has three transposition genes, designated tnpA, tnpB and tnpC. Mutations in each of these were complemented efficiently in trans by clones containing internal fragments of Tn554; thus the products of these genes function in trans. Elements carrying deletions of the Tn554 termini could not be complemented. The product of tnpC is not absolutely required for transposition, since deletion mutations encompassing 80% of tnpC, as well as frameshift mutations located near the amino terminus of tnpC, transposed at frequencies as high as 2% of that observed with wild-type Tn554. However, such mutations affected the orientation of insertion. With wild-type Tn554 insertion occurs in a single orientation regardless of the orientation of the donor. In tnpC mutants insertion orientation was dictated by the orientation of Tn554 in the donor molecule. A mutant lacking the carboxy-terminal 59 residues of tnpB also exhibited altered insertion orientation. Thus it appears that the tnpC gene product is required for correct orientation of the element upon insertion and that this protein may interact with the carboxy-terminal portion of the tnpB gene product.

Blotting, Southern↗
PubMed1988

rpt-1, an intracellular protein from helper/inducer T cells that regulates gene expression of interleukin 2 receptor and human immunodeficiency virus type 1.

The Rpt-1 (for regulatory protein, T-lymphocyte, 1) gene, selectively expressed by resting but not by activated CD4+ inducer T cells, encodes an intracellular protein (rpt-1, Mr 41,000) that down-regulates gene expression directed by the promoter region of the gene encoding interleukin 2 receptor alpha chain and by the long terminal repeat of human immunodeficiency virus type 1. The data reported here suggest that rpt-1 levels may be inversely correlated with activation of CD4+ T cells and human immunodeficiency virus replication leading to clinical symptoms of the acquired immunodeficiency syndrome.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Evidence of glutamatergic denervation and possible abnormal metabolism in Alzheimer's disease.

Excitatory dicarboxylic amino acids previously have been ascribed several functions in the brain. Here their total concentration and proposed neurochemical markers of neurotransmitter function have been measured in brain from patients with Alzheimer's disease (AD) and controls. Specimens were obtained antemortem (biopsy) approximately 3 years after emergence of symptoms and promptly (less than 3 h) postmortem some 10 years after onset. Early in the disease a slight elevation in aspartic acid concentration of cerebral cortex was observed in the patients with AD. A reduction in glutamic acid concentration of a similar magnitude was found. It is argued that this, together with a decrease in CSF glutamine content and lack of change in the phosphate-activated brain glutaminase activity of tissue, reflects an early metabolic abnormality. Later in the disease evidence of glutamatergic neurone loss is provided by the finding that in many regions of the cerebral cortex the Na+-dependent uptake of D-[3H]aspartic acid was almost always lowest in AD subjects compared with control when assessed by a method designed to minimise artifacts and epiphenomena. Release of endogenous neurotransmitters from human brain tissue postmortem did not appear to have the characteristics of that from human tissue antemortem and rat brain.

Aged↗
PubMed1988

Increased mortality rates in late-life depression.

Physical illness at referral and subsequent mortality were studied in a group of elderly patients with depression, and compared with age/sex-matched controls. The depressed group was significantly less well at first interview, and had a significantly higher 4-year mortality. When the effect of physical illness was controlled, the depressed patients (particularly the men) still had a significantly higher 4-year mortality, suggesting that the greater mortality in the depressed group was not due to differences in physical health alone.

Aged↗
PubMed1988

Cell calcium levels of normal and cystic fibrosis nasal epithelium.

To determine whether epithelial ion transport abnormalities in cystic fibrosis (CF) might reflect abnormal regulation of intracellular Ca2+ levels, cytosolic free calcium (Cai2+) was measured using fura-2 or quin2 in suspensions of normal or CF nasal epithelial cells derived from primary cell culture. The basal Cai2+ level measured with fura-2 in CF nasal epithelia was 155 +/- 9 nM (n = 5), a value not significantly different from normal nasal epithelia (143 +/- 16 nM, n = 5). Total cell calcium was measured by atomic absorption spectroscopy and no differences were observed between CF (6.3 +/- 0.5 nmol/mg protein; n = 3) and normal (6.2 +/- 1.2 nmol/mg protein; n = 3) nasal epithelial cells. Placing Na+ loaded cells in a low (10 mM) extracellular Na+ solution resulted in a rapid increase in Cai2+ consistent with Ca2+ uptake via a plasmalemmal Na+-Ca2+ exchanger. The level of Cai2+ achieved by this low Na+ maneuver was not significantly different in CF cells compared to normal cells. Neither isoproterenol (10(-5) M) nor forskolin (10(-6) M) had any effect on Cai2+ in normal or CF nasal epithelial cells. Thus, it appears that differences in cell Cai2+, as measured by fluorescent chelators in suspensions of cultured cells, do not account for the abnormalities in basal or isoproterenol stimulated ion transport in CF tissues.

Adolescent↗
PubMed1988

Ten-year experience with extracorporeal membrane oxygenation for severe respiratory failure.

In the last ten years, 17 patients with respiratory failure refractory to standard ventilator therapy have been treated with extracorporeal membrane oxygenation (ECMO) at the Toronto General Hospital. One patient was treated with ECMO twice. Four perfusions were veno-arterial, the remainder veno-venous. Perfusions ranged from 1 1/2 to 19 days, with a mean of six days. Ten patients died during treatment or soon after it was discontinued. Eight patients improved enough to allow resumption of standard ventilation, and four patients recovered sufficiently to have normal arterial blood gas levels on room air. Three are long-term survivors. Multiple surgical procedures have been performed successfully during use of ECMO including lung lavage, open lung biopsy and three lung transplants. Major complications include hemorrhagic diatheses and sepsis. The technique involves a substantial commitment of time and personnel but remains a tenable option for presumed reversible life-threatening respiratory failure.

Adolescent↗
PubMed1987

Coronary arteriography performed by a physician assistant.

Funding constraints and an oversupply of cardiologists mitigate against continued training of increasing numbers of cardiology fellows. In some institutions, the workload of the catheterization laboratory is an overriding factor. The ability of a physician assistant to perform some of this work was tested to determine if the number of fellows and the content of the fellowship training program could be uncoupled from the catheterization laboratory workload. Among the first 150 patients in whom coronary arteriography was performed by a physician assistant, no patient died or had a myocardial infarction or stroke. Two patients (1.3%) had minor complications: a retinal embolus and an infected puncture site. The complication rate in 150 consecutive cases performed by fellows was also 1.3%, a small myocardial infarction and a transient ischemic attack. Procedure times for the physician assistant and for the fellows were 41 +/- 13 and 44 +/- 18 minutes for preoperative patients and 62 +/- 24 and 70 +/- 20 minutes for postoperative patients. Corresponding fluoroscopy times were 11 +/- 5 and 12 +/- 7 minutes for the preoperative and 22 +/- 12 and 20 +/- 6 for postoperative patients. Only preoperative fluoroscopy times were statistically different (p = 0.02). Thus, substituting a physician assistant for a fellow to perform coronary arteriography is an option in institutions at which the number of studies exceeds the training needs of fellows.

Angiography↗
PubMed1987

In vivo 31P NMR studies of the hepatic response to L-ethionine in anesthetized rats.

Phosphorus-31 surface coil spectroscopy has been used to study the effects of L-ethionine administration on the hepatic metabolism of the anesthetized Sprague-Dawley rat. ATP levels were found to decrease by approximately 30% 3 to 4 hr after administration of 1 mg/g body wt of L-ethionine to the anesthetized rat by gastric gavage. ATP levels returned to control values approximately 8 hr postadministration. The relatively small decrease in ATP level was confirmed by extraction and conventional enzyme assay and is a consequence of the mode of administration of the ethionine. Hepatic inorganic phosphate levels rose concomitantly with the ATP fall. There were no significant changes in either cellular pH or Mg2+ levels as monitored by the 31P shifts of sensitive metabolites. In vivo 31P NMR spectroscopy provides a promising approach to study the effects of hepatotoxicants on cellular ATP, pH, and Mg levels.

Adenosine Triphosphate↗