[Evaluation of cardiac function by radioisotope angiography].

Cardiac function can be assessed with noninvasive radionuclide technique. Radioisotope angiography can be performed either using equilibrium acquisition or first pass technique. The most commonly generated measurements are as follows; 1) chamber-to-chamber transit time, 2) cardiac output, 3) left-to-right shunt ratio, 4) left and right ventricular ejection fractions, 5) left ventricular end-diastolic and end-systolic volumes, 6) regional wall motion, 7) peak ejection rate, and 8) peak filling rate.

Lysophosphatidylcholine triggers intracellular calcium release and activation of non-selective cation channels in renal arterial smooth muscle cells.

The effects of a lipid component of oxidized low-density lipoproteins (ox-LDL), L-alpha-palmitoyl-lysophosphatidylcholine (LPC), on membrane currents of isolated canine renal artery smooth muscle cells (RASMC) were examined using the whole-cell configuration of the patch-clamp technique. In RASMC exposed to nominally Ca2+-free solutions and dialyzed with 0.1 mM EGTA and 140 mM K+, superfusion with LPC (10 microM) elicited spontaneous transient outward currents (STOCs) and/or spontaneous transient inward currents (STICs), followed by the activation of a large voltage-independent current with a reversal potential (Er) close to 0 mV.

Protective roles of endogenous carbon monoxide in neointimal development elicited by arterial injury.

We reported that carbon monoxide (CO) generated through heme oxygenase (HO) inhibits mitogen-induced proliferation of vascular smooth muscle cells (VSMCs). We report that balloon injury induces HO-1, the stress-inducible isozyme of HO, in VSMCs and inhibits neointimal formation through the action of endogenous CO. Northern blot analysis and immunohistochemistry revealed that HO-1 is markedly induced in the media as early as 1 day after injury, whereas only a little expression was detected in the intact carotid artery.

A 38 kb segment containing the cdc2 gene from the left arm of fission yeast chromosome II: sequence analysis and characterization of the genomic DNA and cDNAs encoded on the segment.

A genomic 38 kbp segment on the c1750 cosmid clone containing the cdc2 gene, located in the left arm of chromosome II from Schizosaccharomyces pombe, was sequenced. The segment was found to have five previously known genes, pht1, cdc2, his3, act1 and mei4. Among 11 coding sequences (CDSs) predicted by the gene finding software INTRON.

Anion transport in heart.

Anion transport proteins in mammalian cells participate in a wide variety of cell and intracellular organelle functions, including regulation of electrical activity, pH, volume, and the transport of osmolites and metabolites, and may even play a role in the control of immunological responses, cell migration, cell proliferation, and differentiation. Although significant progress over the past decade has been achieved in understanding electrogenic and electroneutral anion transport proteins in sarcolemmal and intracellular membranes, information on the molecular nature and physiological significance of many of these proteins, especially in the heart, is incomplete. Functional and molecular studies presently suggest that four primary types of sarcolemmal anion channels are expressed in cardiac cells: channels regulated by protein kinase A (PKA), protein kinase C, and purinergic receptors (I(Cl.

Purinoceptor-coupled Cl- channels in mouse heart: a novel, alternative pathway for CFTR regulation.

1. P2-purinoceptors couple extracellular ATP to the activation of a Cl- current (ICl,ATP) in heart. We studied the molecular mechanism and intracellular signalling pathways of ICl,ATP activation in mouse heart.

Two cases of methotrexate induced lymphomas in rheumatoid arthritis: an association with increased serum IgE.

We describe 2 patients with rheumatoid arthritis (RA) in whom non-Hodgkin's lymphomas developed during low dose pulsed methotrexate (MTX) treatment. The tumors regressed after discontinuation of MTX with no additional treatment. Serum levels of IgE increased concomitantly with the development of lymphoma, and decreased along with the regression of the lymphoma in both patients.

Complete genome sequence of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1.

The complete sequence of the genome of an aerobic hyper-thermophilic crenarchaeon, Aeropyrum pernix K1, which optimally grows at 95 degrees C, has been determined by the whole genome shotgun method with some modifications. The entire length of the genome was 1,669,695 bp. The authenticity of the entire sequence was supported by restriction analysis of long PCR products, which were directly amplified from the genomic DNA.

Determination of concentrations of flecainide in human serum by high-performance liquid chromatography on a fluorocarbon-bonded silica gel column.

An optimized method for the determination of flecainide in serum is presented. Extraction using a solid-phase C18 column and chromatography on a stabilized fluorocarbon-bonded silica gel column effectively separate flecainide from an internal standard (a positional isomer of flecainide). The HPLC apparatus and conditions were as follows: analytical column, Fluofix 120N; sample solvent, 20 microl; column temperature, 40 degrees C; detector, Shimadzu RF-5000 fluorescence spectrophotometer (excitation wavelength = 300 nm, emission wavelength = 370 nm); mobile phase, 0.

Temporal differences in actions of calcium channel blockers on K+ accumulation, cardiac function, and high-energy phosphate levels in ischemic guinea pig hearts.

We investigated temporal differences in the protective action of three types of Ca2+ channel blockers in myocardial ischemia, focusing particularly on the blocking ability under depolarizing conditions. The effects of diltiazem, verapamil, and nifedipine on extracellular potassium concentration ([K+]e), acidosis, and level of metabolic markers were examined during 30-min global ischemia and postischemic left ventricular (LV) function in isolated guinea pig hearts. Diltiazem and verapamil, but not nifedipine, inhibited the late phase (15-30 min) of [K+]e elevation, whereas all three blockers delayed the onset of the early phase (0-8 min) of [K+]e elevation.

Regulation of recombinant cardiac cystic fibrosis transmembrane conductance regulator chloride channels by protein kinase C.

We investigated the regulation of cardiac cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channels by protein kinase C (PKC) in Xenopus oocytes injected with cRNA encoding the cardiac (exon 5-) CFTR Cl- channel isoform. Membrane currents were recorded using a two-electrode voltage clamp technique. Activators of PKC or a cAMP cocktail elicited robust time-independent Cl- currents in cardiac CFTR-injected oocytes, but not in control water-injected oocytes.

Identification of a new glycosylphosphatidylinositol-anchored 42-kDa protein and its C-terminal peptides from bovine erythrocytes by gas chromatography-, time-of-flight-, and electrospray-ionization-mass spectrometry.

By treatment with phosphatidylinositol-specific phospholipase C (PIPLC), we obtained several candidates of glycosylphosphatidylinositol (GPI)-anchored proteins such as 55, 42, 40, and 30 kDa from bovine erythrocyte membrane, in addition to the well-known GPI-anchored protein acetylcholinesterase. In these proteins, the presence of myo-inositol was confirmed by gas chromatography (GC)-mass spectrometry. Among them, the 42-kDa protein was further analyzed by electrospray-ionization (ESI)-mass spectrometry (MS) after hydrolysis by lysyl endoprotease.

Biphasic regulation of the development of murine type II collagen-induced arthritis by interleukin-12: possible involvement of endogenous interleukin-10 and tumor necrosis factor alpha.

Objective: To examine the dose-specific effects of interleukin-12 (IL-12) on the evolution of murine type II collagen-induced arthritis (CIA).

Methods: From day 24 through day 33 following primary immunization, mice received daily intraperitoneal injections of murine recombinant IL-12. Measurements of anticollagen IgG, cytokines, and corticosterone were performed using enzyme-linked immunosorbent assay and radioimmunoassay.

Real time measurement of nitric oxide released from cultured endothelial cells.

Direct detection of nitric oxide (NO) is essential for understanding the precise mechanism of its production from endothelial cells. Previously, we developed an NO detection system based on the chemiluminescence reaction between NO and luminol-H2O2. Here, we have applied this system to cultured endothelial cells for the direct and on-time measurement of NO.

[Use of 99mTc-labeled myocardial perfusion agents in the Kanto-Koshinetsu area].

We sent out a questionnaire to investigate how the 99mTc-labeled myocardial perfusion agents are used in the Kanto-Koshinetsu area. Answers were obtained from 83 laboratories. Examinations using 99mTc-labeled agents were performed more frequently than those using 201T1 only in 10 of 83 laboratories.

[Clinical significance of 123I-BMIPP myocardial SPECT].

123I-BMIPP is a radioiodinated branched chain fatty acid analog, showing high accumulation as well as prolonged retention in the myocardium. Therefore, it is used as a metabolic imaging agent suitable for myocardial SPECT. After 123I-BMIPP is taken in the myocardium, it remains there mainly as a storage type fatty acid of triglyceride, and it shows different behavior by the stage of cardiac disease.

[Pirarubicin-induced myocardial damage in elderly patients with non-Hodgkin's lymphoma].

123I-metaiodobenzylguanidine (MIBG) myocardial single photon emission computed tomography (SPECT), 123I-beta-methyliodophenyl pentadecanoic acid (BMIPP) myocardial SPECT, and holter ECG recording were performed in patients with non-Hodgkin's lymphoma who underwent chemotherapy including pirarubicin (THP), in an attempt at early detection of cardiac toxicity from THP. Twenty-six patients with untreated non-Hodgkin's lymphoma who received THP-COPBLM therapy were studied. For THP-COPBLM therapy.

Direct activation of endothelial NO pathway by Ba2+ in canine coronary artery.

1. We have reported that Ba2+ causes endothelium-dependent relaxation of canine coronary arteries through NO synthesis in Ca2+-free and depolarizing solution. To determine the cellular mechanisms by which the endothelium-dependent relaxation occurs, we used fura-2 fluorometry (F350 and F390; excitation wavelengths, 350 and 390 nm, respectively) and estimated the intracellular Ba2+ concentration in endothelial and vascular smooth muscle cells.

Shortening of action potential duration is not prerequisite for cardiac protection by ischemic preconditioning or a KATP channel opener.

Our purpose was to determine whether amelioration of myocardial contractile dysfunction by an ATP-sensitive potassium (KATP) channel opener or ischemic preconditioning is mediated by shortening of action potential duration during the early period of ischemia. Extracellular potassium concentration ([K+]e), monophasic action potential duration (MAPD) and thickening fraction were measured during ischemia and reperfusion of the left anterior descending coronary artery in anesthetized dogs. Control dogs were subjected to a 10-min occlusion (test occlusion) followed by a 120-min reperfusion.

Complete sequence and gene organization of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3.

The complete sequence of the genome of a hyper-thermophilic archaebacterium, Pyrococcus horikoshii OT3, has been determined by assembling the sequences of the physical map-based contigs of fosmid clones and of long polymerase chain reaction (PCR) products which were used for gap-filling. The entire length of the genome was 1,738,505 bp. The authenticity of the entire genome sequence was supported by restriction analysis of long PCR products, which were directly amplified from the genomic DNA.

5-hydroxydecanoate selectively reduces the initial increase in extracellular K+ in ischemic guinea-pig heart.

The aim of the present study was to determine the effect of 5-hydroxydecanoate (5-HD) on extracellular K+ levels during global ischemia for 30 min employing K+-sensitive electrodes in isolated guinea-pig hearts. 5-HD (100 microM) reduced the K+ accumulation during the early period of ischemia, but did not inhibit the elevation of extracellular K+ in the latter half of the ischemic period which was selectively enhanced by ouabain (3 microM). Thus, 5-HD appears to exert a similar mode of action as glibenclamide on extracellular K+ accumulation in the ischemic guinea-pig hearts.

Functional and molecular expression of volume-regulated chloride channels in canine vascular smooth muscle cells.

1. We examined the possibility of functional and molecular expression of volume-regulated Cl- channels in vascular smooth muscle using the whole-cell patch-clamp technique and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) on cells from canine pulmonary and renal arteries. 2.

Purification and amino acid sequence of halystase from snake venom of Agkistrodon halys blomhoffii, a serine protease that cleaves specifically fibrinogen and kininogen.

We have isolated a serine protease, halystase, from Agkistrodon halys blomhoffii venom by chromatography on DEAE-Sepharose, heparin-Sepharose and Q-Sepharose columns, and have determined the complete amino acid sequence by Edman degradation and by mass spectral analysis of peptides generated by enzymatic and chemical cleavage. The 238-residue sequence of halystase, containing N-linked carbohydrates (about 13%) at two sites showed significant similarity to other thrombin-like snake venom serine proteases (66-72%), mammalian tissue kallikrein (42%) and thrombin (26%). Halystase contained the tentative catalytic triad of His43, Asp88 and Ser184 common to all serine proteases and Asp178 in the primary substrate-binding site.

Shortening of monophasic action potential duration during hyperkalemia and myocardial ischemia in anesthetized dogs.

The elevation of the myocardial extracellular potassium concentration ([K+]o) is known to shorten action potential duration, which may lead to the occurrence of arrhythmias. The aim of this study was to compare the mechanisms responsible for the shortening of monophasic action potential duration (MAPD) in hyperkalemic and myocardial ischemic hearts in anesthetized dogs. During a venous infusion of KCl for 5 min, [K+]o was increased and MAPD was significantly shortened.