The value of electrocardiographic R-wave changes in exercise testing: Preexercise versus postexercise measurements

To determine the usefulness of R-wave amplitude changes during exercise testing for the diagnosis of coronary artery disease (CAD) and to understand the discrepancies that have been described in the literature regarding their value, we studied two groups of patients by means of electrocardiographic (EKG) treadmill testing and coronary arteriography. Group I was composed of 149 patients who were studied prospectively. The specificity of R-wave changes measured from preexercise to immediately postexercise (SRV(5)) was 81%, but that of R-wave changes measured from preexercise to peak exercise (URV(5)) was 46%. A group of 156 patients (Group II) evaluated retrospectively showed a high specificity for the SRV(5) (84%) and poor specificity for the URV(5) (39%). The sensitivity of the SRV(5) was 38% in Group I and 42% in Group II. Therefore, if measured during the immediate postexercise period and not at peak exercise, changes in R-wave amplitude may be of value in the diagnosis of coronary artery disease by electrocardiographic exercise testing.     

Pineal cells enhance choline acetyltransferase activity in sympathetic neurons

Cells derived from the neonatal rat pineal gland were cocultured with cells derived from neonatal rat superior cervical ganglia (SCG) in an attempt to determine whether a sympathetic target organ with only adrenergic properties could enhance the development of adrenergic transmitter properties in sympathetic neurons in tissue culture. Choline acetyltransferase was measured as an index of cholinergic differentiation, and tyrosine hydroxylase was measured as an index of adrenergic differentiation. As indices of total cell number and cellular volume, DNA and protein, respectively, were also measured. We found that the pineal-SCG cocultures contained ten times greater choline acetyltransferase activity than sister neuronal cultures cultured without pineal cells, thus indicating that the pineal cells enhanced cholinergic properties in the sympathetic neurons. This cholinergic enhancement was dependent upon the presence of nerve growth factor and could not be obtained with pineal-conditioned medium. Tyrosine hydroxylase activity, measured on cultures sister to those mentioned above, was low in all cultures and decreased somewhat in SCGs cultured alone. TH activity in the pineal-SCG cocultures, however, increased slightly. Some tyrosine hydroxylating activity developed in pineals cultured alone, however, and may have been responsible for the small increase in tyrosine hydroxylase activity noted in the pineal-SCG cocultures. The implications of these results for a determination of the role that target organ plays in the development of the transmitter properties of sympathetic neurons are discussed.     

Isolation and characterization of mutants ofEscherichia coli defective in pyridine nucleotide cycle enzymes

Thirty-eight analogs of nicotinic acid and nicotinamide were tested for their ability to inhibit growth of wild-typeEscherichia coli K-12. Two of the compounds tested, 6-aminonicotinic acid and 6-aminonicotinamide were strongly inhibitory to growth of the organism. Mutants resistant to these compounds were isolated and characterized by cross-feeding experiments. All of the mutants isolated by their resistance to these analogs were found to excrete a metabolite which supported growth ofnadA, nadB, ornadC strains ofE. coli on a minimal medium. Wild-type strains failed to exhibit this cross-feeding ability. ThepncB ⁺ locus codes for nicotinic acid phosphoribosyl transferase and maps near minute 23 on the chromosome.     

Plasmids coding for heat-labile enterotoxin production isolated from Escherichia coli O78: comparison of properties.

Nineteen enterotoxigenic Escherichia coli strains of serogroup O78, isolated in different geographical areas from humans with diarrheal diseases, were tested for their ability to transfer enterotoxin production. All of the strains originally produced heat-labile enterotoxin, and 16 also produced heat-stable enterotoxin and colonization factor antigen I. Plasmids coding for the production of heat-labile enterotoxin only were transferred from 13 strains. Some properties of these plasmids were compared. All were fi+, but they could be divided into three groups on the basis of their incompatibility reactions, ability to restrict E. coli K-12 phages, and size. The three heat-labile enterotoxin plasmids isolated from African strains all belonged to one enterotoxin plasmid group. The heat-labile enterotoxin plasmids from the Asian strains were divided into two groups, those from serotype O78.H11 differing from those from serotype O78.H12.     

Cell wall characteristics of Pseudomonas aeruginosa and its carbenicillin-induced L-form.

L-forms of Pseudomonas aeruginosa were induced and cultured on a medium supplemented with carbenicillin. Morphological studies of the passaged variant revealed the presence of a triple-layered cell wall similar to that found in the parent species. Furthermore, the L-form was found to be more susceptible to gentamicin, kanamycin, tetracycline and colistin sulphate. Chemical analysis of the lipopolysaccharide fraction showed a difference in phosphorus content, and changes in cell wall envelope fatty acid content were also exhibited. It is suggested that these differences may influence the transport of certain antibiotics through the cell wall.     

Cyclic AMP metabolism in mouse parotid glands. Properties of adenylate cyclase, protein kinase and phosphodiesterase.

Catecholamines induce unique growth and secretory responses in salivary glands. An analysis of three enzyme activities involved in cyclic AMP metabolism was carried out to identify the specificity of these responses for salivary glands. Although parotid adenylate cyclase has an unusually high specific activity, its kinetic properties and responses to NaF, guanine nucleotides, and isoproterenol are similar to other tissues not stimulated to grow after isoproterenol stimulation. Solubilized adenylate cyclase was separated from other membrane proteins by isoelectric focusing on polyacrylamide gels. There was a single broad peak of activity witha pI of 5.9. Parotid protein kinase has a subcellular distribution and substrate preference similar to hepatic protein kinase. Activation by cyclic AMP is also similar to that reported for other tissues, with a Ka of 1.2 - 10(-7) M. Parotid cyclic AMP and cyclic GMP phosphodiesterases are a heterogeneous group of enzymes with relatively low specific activity as compared with mouse pancreas, liver and brain. Isoelectric focusing of supernatant phosphodiesterases revealed at least sixpeaks of enzyme activity in the pI range of 4-6. Previous reports of a large increase in parotid cyclic AMP levels after in vivo administration of catecholamines and specific growth and secretion could be the result of a relatively high specific activity adenylate cyclase associated with low specific activity cyclic AMP phosphodiesterases.     

PHARMACOLOGICAL STUDIES ON THE STIMULATION OF THE PHOSPHOLIPASE A2-ACYLATION SYSTEM OF SYNAPTIC MEMBRANES OF BRAIN, BY NEUROTRANSMITTERS AND OTHER AGONISTS

Methacholine, nicotine and succinylcholine stimulated the phospholipase A2-acylation system of synaptic membranes isolated from the cerebral cortex of guinea pig. Stimulation by acetylcholine was partially blocked by atropine and by D-tuberocurarine respectively, indicating both muscarinic and nicotinic stimulation. Muscarinic stimulation by acetylcholine was greater than -isotinic stimulation, and stimulation by acetylcholine was completely blocked by a combip, or;. and n-tuberocurarine. The phospholipase A2-acylat tem was stimulated by phenylcphrine., id. Cqxoterenoi. Stimulation by noradrenaline was J-. tidlr, by phenoxybenzamine and pindalol i:spectively, indicating both 8-adrenergic and P-adrenergic ztimulation. n-Adrenergic stimulation by noradrenaline was greater than P-adrenergic-stimulation. 5 -mlation by noradrenaline was completely blocked by a combination of phenoxybenzamins and pindalol. Stimulation of both acylation and phospholipid hydrolysis, by 5-hydroxytryptamine and histamine were partially blocked by methysergide and diphenhydramine respectively. Stimulation by dopamine was blocked by halopcridol. Stimulation by y-aminobutyric acid was partially blocked by strychnine and by picrotoxin. Dichloroisoproterenol, atropine, methysergidr, diphenhydramine, strychnine, picrotoxin and eserine, at relatively high concentrations (1 mM), stimuhted the phospholipase A2-acylation system. Synergistic stimulations of both acylatior, and hydrolysis of phosphatidylcholine, were observed by adenosine combined with noradrenaline, 5-hydroxytryptamine, histamine, dopamine or yaminobutyric acid, respectively. In the presence of ATP-MgCI, synergistic stimulations of the hydrolysis of phosphatidyicholine were observed after 30 s by noradrenaline combined with 5-hydroxytryptamine, histamine, dopamine, aminobutyric acid or carbamoylcholine respectively. In the presence of GTP-MgC12 synergistic stimulations were obtained by cdrbamoylcholine combined with noradrenaline. 5-hydroxytryptamine. histamine, dopamine or y-aminobutyric acid, respectively. In the presence of ATP-MgC12 plus GTP-MgC12, stimulation by noradrenaline and one other agonist including 5-hydroxytryptamine, histamine, dopamine, y-aminobutyric acid or carbamoylcholine were close to additive.     

The stimulation of the phospholipase A2-acylation system of synaptic membranes of brain by cyclic nucleotides.

Hydrolysis of phosphatidylcholine by phospholipase A2 of synaptic membranes i n Tris-CHl buffer was stimulated by cyclic AMP, cyclic GMP, cyclic CMP, cyclic UMP and adenosine (0.1 mm). In the presence of 1 mm-NaF and cofactors, the same cyclic nucleotides and adenosine (10 mm) stimulated the incorporation of added oleate into the choline glycerophospholipids of synaptic membranes. Cyclic AMP and noradrenaline stimulated the incorporation of added oleate into position 2 of choline glycerophospholipid. Stimulation of net acylation was increased by preincubation in conditions which stimulated hydrolysis of phosphatidylcholine. Cyclic AMP only slightly stimulated the transfer of oleate from oleoyl-CoA into choline glycerophospholipid. The optimum concentration of CaCl2 for the stimulation of hydrolysis by phospholipase A2 by cyclic AMP was 1 mum. Stimulation of the incorporation of added oleate was maximal in the CaCl2 concentration range 1 mum-1mm. MgCl2 also enhanced stimulations, maximum effects being obtained with concentrations of 10 mum and 0.5 mm for hydrolysis by phospholipase A2 and incorporation of added oleate respectively. ATP enhanced the stimulation of incorporation of oleate but had no effect on the cyclic nucleotide stimulation of hydrolysis of added phosphatidylcholine by phospholipase A2. Adenosine, guanosine, ADP and 5'-AMP (all at 1 mm) inhibited the stimulation of incorporation of oleate by cyclic nucleotides and inhibited the transfer of oleate from oleoyl-CoA to phospholipid. They did not inhibit the stimulation of hydrolysis of added phosphatidylcholine (by phospholipase A2) by cyclic nucleotides, but inhibited the stimulation by noradrenaline, acetylcholine, 5-hydroxytryptamine, dopamine (3,4-dihydroxyphenethylamine) and histamine. Preincubation of synaptic membranes in the water or buffer increased the net activity of phospholipase A2. Preincubation with a mixture of ATP and MgCl2 increased the initial rate of acylation of membrane lipid.