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.     

Isolation of selective 3H-chlornaltrexamine-bound complexes,possible opioid receptor components in brains of mice

The nonequilibrium narcotic antagonist, chlornaltrexamine (CNA) was used to bind selectively and covalently pioid specific sites on brain membrane preparations. Selective binding of [³H]CNA occured with a saturation maximum of 185 fmol/mg protein. Bound [³H]CNA was extracted with Triton X-100, dialyzed against Brij 36T, precipitated with trichloroacetic acid and chromatographed on an ultrogel AcA 22 column. The elution profile suggests that this extract contains a minimum of four selective [³H]CNA complexes. At least two of these complexes migrate in a single large peak. Column calibration showed that this peak eluted at 590,000 daltons. One of these specific [³H]CNA complexes elutes at the elution volume of the column and is dialyzable. Finally, putative aggregate of these complexes elutes with the void volume.     

Lipoxygenase from cucumber fruit: Localization and properties

The subcellular localization of lipoxygenase (LOX) from cucumber fruit has been studied. Two methods have been employed to obtain organelles; (1) maceration of the tissue, followed by separation on a linear sucrose gradient and (2) release from protoplasts by osmotic shock, followed by a discontinuous Ficoll gradient. It was possible to obtain high LOX activity in the intact protoplasts from both peel and flesh tissue. However, fewer intact vacuoles were obtained following osmotic rupture than from macerated tissue. Both methods produced more particulate LOX activity from the peel than from flesh tissue, and both showed that this activity was associated with the vacuoles. The cucumber LOX enzyme was similar to the potato and tomato enzymes, both in pH characteristics and substrate specificity.     

Development of transmitter secretory mechanisms by adrenergic neurons in the embryonic chick heart ventricle

Developmental changes in the function of adrenergic axons within the right ventricle of the chick embryo were assessed by measuring the ability of these axons (1) to release endogenous transmitter, and (2) to transport, retain, and release tritiated norepinephrine ([³H]NE). The release of endogenous catecholamines was assayed indirectly by measuring the increase in the twitch tension of ventricular muscle evoked by electrical stimulation of intramural nerves. The release of endogenous transmitter, which acted via β-adrenergic receptors, was first detected by this method on the 16th embryonic day. A cocaine-sensitive uptake of [³H]NE was first observed on the 12th embryonic day. At this time, elevated potassium first evoked a calcium-sensitive release of [³H]NE. Electrical stimulation of intramural axons first evoked a tetrodotoxin-sensitive release of [³H]NE on the 14th embryonic day. It is concluded that the axons of developing adrenergic neurons are capable of releasing transmitter soon after they contact their target tissue.     

Mixed micelles of sphingomyelin and phosphatidylcholine with nonionic surfactants. Effect of temperature and surfactant polydispersity.

Mixed micelle formation of the polydisperse nonionic surfactant Triton X-100 as well as its homogeneous analogue, p-(1,1,3,3-tetramethylbutyl)-phenoxynonaoxyethylene glycol (OPE-9), with bovine brain sphingomyelin or dipalmitoyl phosphatidylcholine has been characterized by column chromatography on 6% agarose. At 40 degrees C, mixtures of OPE-9 and either sphingomyelin or dipalmitoyl phosphatidylcholine give a narrow size distribution for mixed micelles. A this temperature the size distribution of Triton X-100-containing mixed micelles is complicated because of the polydispersity of the oxyethylene chains. At 20 degrees C narrow size distributions are observed for mixed micelles of sphingomyelin/Triton X-100 and sphingomyelin/OPE-9 up to at least 0.06 mol fraction of lipid. For dipalmitoyl phosphatidylcholine this is observed only with OPE-9. At intermediate mol fractions of lipid (around 0.25), two populations of mixed micelles exist for sphingomyelin/Trition X-100, sphingomyelin/OPE-9, and dipalmitoyl phosphatidylcholine/OPE-9. At high mol fractions of lipid only one population of mixed micelles again exists. At 20 degrees C, sphingoymelin forms a clear solution with Triton X-100 and OPE-9 to a lipid mol fraction of at least 0.46 and 0.67, respectively. Dipalmitoyl phosphatidylcholine forms a clear solution with OPE-9 to a lipid mol fraction of at least 0.57 at the same temperature. Triton X-100 and dipalmitoyl phosphatidylcholine do not form stable, clear solutions at 20 degrees C unless the lipid mol fraction is extremely low. These results show that surfactant polydispersity and temperature are important determinants in the solubilization of lipids by nonionic surfactants. It is also shown that pure surfactant micelles and lipid/surfactant mixed micelles do not co-exist in the same solution.     

Action of cobra venom phospholipase A2 on the gel and liquid crystalline states of dimyristoyl and dipalmitoyl phosphatidylcholine vesicles

The activity of phospholipase A2 from cobra venom toward phospholipid in single-walled, sonicated vesicles was analyzed, particularly with respect to its activity toward the saturated phosphatidylcholines in the gel and liquid crystalline states. When egg phosphatidylcholine vesicles are used as substrate, the phospholipase has an apparent Km of 4.4 mM, an apparent Vmax of 100 mumol min-1 mg-1 of protein, and a pH optimum of 5.0 at 40 degrees C. The phospholipase hydrolyzed the gel state of dimyristoyl phosphatidylcholine vesicles and dipalmitoyl phosphatidylcholine vesicles at a rate 2 to 3 times greater than the liquid crystalline state, taking into account temperature effects on the enzymatic reaction itself. The results suggest that, toward sonicated vesicles, there is no specific enhancement of the rate when the both liquid crystalline and gel states are present together, as has been suggested to occur for multibilayers studied with other phospholipases. An apparent stimulation of activity as the reaction proceeded was observed above the phase transition temperature. This might be attributed to an increase in the phase transition temperature caused by free fatty acids so that, in the presence of reaction products, the enzyme is actually hydrolyzing gel state phospholipid which was found to be the preferred lipid state for phospholipase activity.     

A rapid and sensitive method for the detection of histone peptides

Fluorescamine has been used to obtain a peptide map of a mixture of histones (H₃, H₂A, H₂B, and H₄) prepared from oocytes of Xenopus laevis. Fluorescamine was found to be more sensitive than o-phthalaldehyde or ninhydrin-Cd for the detection of peptide fragments obtained from tryptic digestion of oocyte histones of X. laevis and the peptic digestion of the β chain of insulin. Using the β chain of insulin for a comparison, the 8 major peptide fragments could be separated by electrophoresis within 30 min and were detectable at the picomols level. Some 70 peptide spots of X. laevis oocyte histones were resolved, thus permitting the analysis of this complex mixture of polypeptides without the need for prior separation.