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Biomedical subjects

M Hatano

Publications and source records attributed to M Hatano.

At least 217 records · Page 12Linked to original sources

Isolation of apolipoproteins from carotenoid-carrying lipoprotein in the serum of chum salmon, Oncorhynchus keta.

Carotenoid-carrying lipoprotein (CCL) was rapidly isolated from the high density lipoprotein (HDL) fraction of the upstream migrating male chum salmon (Oncorhynchus keta) by a single-step density gradient ultracentrifugation. The two apolipoproteins (Mr = 24,000 and 12,000; designated apo-I and apo-II, respectively) were readily dissociated and separated in 0.1% SDS by gel filtration chromatography. Prominent features of the amino acid composition in the CCL included the relative high levels of glutamic acid, alanine, leucine, and lysine, and the low cysteine content. Apo-I, as well as the CCL, was rich in glutamic acid, alanine, leucine, and lysine. Compared to the amino acid composition of apo-I, apo-II included relatively high levels of glycine and tyrosine, and low threonine, serine, and arginine contents. When the intact CCL particle was treated with trypsin, apo-I was rapidly proteolyzed, while apo-II was resistant. However, both apo-I and apo-II isolated from the CCL particle were readily digested with trypsin. This suggested that a different structural arrangement rather than the amino acid compositions of the apolipoproteins was associated with the limited trypsin digestion of the CCL particle. Apo-II may be sheltered from the aqueous environment and lie partly within the CCL particle. The properties of both the HDL fraction and apolipoproteins from pink salmon (Oncorhynchus gorbuscha) were similar to those of the CCL from chum salmon.

Amino Acids↗

Production of C3 nephritic factor by cultured lymphocytes derived from a patient with partial lipodystrophy.

C3 nephritic factor (C3 NeF) has been found mainly in the sera of patients with membranoproliferative glomerulonephritis (MPGN) and partial lipodystrophy (PLD). We examined whether peripheral blood mononuclear cells (PBMC) from a patient with PLD could produce C3 NeF. We investigated the in vitro immunoglobulin synthesis of PBMC with mitogen. We further studied the C3bBb stabilizing activity and undertook agglutination assays of the IgG obtained from the culture supernatants. The patient IgG was able to agglutinate only EAC4b3bBb cells and none of the other intermediate cells. We this demonstrated that C3 NeF could be produced in vitro by PBMC derived from a patient with PLD.

Child↗

[Four cases of mitral valve prolapse associated with ventricular tachycardia].

Four patients with mitral valve prolapse and ventricular tachycardia or fibrillation were described. Case 1 was a 46-year-old man with syncope due to ventricular fibrillation. Case 2 was a 36-year-old woman with palpitation and dizziness due to ventricular tachycardia. Case 3 was a 47-year-old man with palpitation and dizziness due to ventricular tachycardia. Case 4 was a 38-year-old man with syncope due to torsade de pointes. Left ventriculograms showed mitral valve prolapse in all cases. Coronary arteriograms were also normal, supporting a noncoronary etiology of the arrhythmia. Patients with unexplained ventricular arrhythmias should be screened for mitral valve prolapse.

Adult↗

Sequence of mechanical and electrical changes during myocardial ischemia: assessment by an ambulatory left ventricular function monitor.

To investigate the relationship between left ventricular function and electrical changes during myocardial ischemia, ambulatory left ventricular function monitoring and ECG recording were made during the ergometer exercise test in 14 patients with coronary artery disease. An ambulatory ventricular function monitor consists of a small cadmium telluride (CdTe) radionuclide probe (250 g) affixed to the patient's chest wall, a preamplifer (10 g), and a portable data acquisition unit (600 g). Left ventricular time-activity curves were recorded continuously using this monitor, and the end-systolic count (volume), end-diastolic count (volume) and ejection fraction were calculated after background subtraction. Twenty-eight exercise tests were performed in the supine and upright positions. In 15 tests, left ventricular dysfunction, i.e., an increase in the end-systolic count (greater than or equal to 10%) and a decrease in ejection fraction (greater than or equal to 5%), and ST depression (greater than or equal to 0.1 mV) were observed. In these 15 tests, exercise duration was 362 +/- 27 sec. Left ventricular dysfunction occurred earlier than ST depression and the time difference was 97 +/- 19 sec. Left ventricular function recovered 33 +/- 8.5 sec after discontinuation of exercise, while ST depression continued for the additional 85 +/- 18.5 sec after recovery of left ventricular function. In conclusion, 1) left ventricular dysfunction occurs earlier than electrical changes during exercise-induced ischemia; 2) left ventricular dysfunction improves earlier than electrical changes after exercise; and 3) the same temporal sequence exists in the restoration from myocardial ischemia.

Coronary Disease↗

Properties of the reaction center of the thermophilic purple photosynthetic bacterium Chromatium tepidum.

Reaction centers were purified from the thermophilic purple sulfur photosynthetic bacterium Chromatium tepidum. The reaction center consists of four polypeptides L, M, H and C, whose apparent molecular masses were determined to be 25, 30, 34 and 44 kDa, respectively, by polyacrylamide gel electrophoresis. The heaviest peptide corresponds to tightly bound cytochrome. The tightly bound cytochrome c contains two types of heme, high-potential c-556 and low-potential c-553. The low-potential heme is able to be photooxidized at 77 K. The reaction center exhibits laser-flash-induced absorption changes and circular dichroism spectra similar to those observed in other purple photosynthetic bacteria. Whole cells contain both ubiquinone and menaquinone. Reaction centers contain only a single active quinone; chemical analysis showed this to be menaquinone. Reaction center complexes without the tightly bound cytochrome were also prepared. The near-infrared pigment absorption bands are red-shifted in reaction centers with cytochrome compared to those without cytochrome.

Bacterial Proteins↗

Proton and iodine-127 nuclear magnetic resonance studies on the binding of iodide by lactoperoxidase.

Interaction of an iodide ion with lactoperoxidase was studied by the use of 1H NMR, 127I NMR, and optical difference spectrum techniques. 1H NMR spectra demonstrated that a major broad hyperfine-shifted signal at about 60 ppm, which is ascribed to the heme peripheral methyl protons, was shifted toward high field by adding KI, indicating the binding of iodide to the active site of the enzyme; the dissociation constant was estimated to be 38 mM at pH 6.1. The binding was further detected by 127I NMR, showing no competition with cyanide. Both 1H NMR and 127I NMR revealed that the binding of iodide to the enzyme is facilitated by the protonation of an ionizable group with a pKa value of 6.0-6.8, which is presumably the distal histidyl residue. Optical difference spectra showed that the binding of an aromatic donor molecule to the enzyme is slightly but distinctly affected by adding KI. On the basis of these results, it was suggested that an iodide ion binds to lactoperoxidase outside the heme crevice but at the position close enough to interact with the distal histidyl residue which possibly mediates electron transport in the iodide oxidation reaction.

Horseradish Peroxidase↗

Host-dependent temperature-sensitive growth of HVJ (Sendai virus) wild-type in rat glioma C 6 cells.

At non-permissive temperature viral specific RNA synthesis was not restricted in rat glioma (C 6) cells infected with HVJ (Sendai virus) wild-type. However, as has previously been shown (J Gen Virol [1984] 65: 639-643), the synthesis of M protein was reduced at non-permissive temperature, in contrast to the L, P, HN, Fo and NP proteins which were synthesized in comparable amounts at permissive and non-permissive temperatures. In this report we show additionally that viral nucleocapsids (NC), which consist of L, P and NP proteins, were formed within the infected cells at both temperatures. Hemagglutinin and neuraminidase activities were also detected in samples incubated at non-permissive temperature. By membrane immunofluorescence and cell-surface immunoprecipitation it was shown that migration of HN and Fo proteins to the cell surface occurred normally at non-permissive temperature. Additionally, the L, P and NP proteins, which were associated with the plasma membrane isolated from the infected cells maintained at permissive temperature, were absent from the membrane of cells incubated at non-permissive temperature. These results suggest that NC and glycoproteins synthesized at non-permissive temperature could not assemble effectively at the plasma membrane because of a lack of M protein. Thus, the host-dependent ts lesion of HVJ in C 6 cells was considered to be mainly in M protein synthesis.

Animals↗

Establishment and biological characterization of an in vitro human cytomegalovirus latency model.

In an attempt to develop an in vitro human cytomegalovirus (HCMV) latency model system, the growth characteristics of HCMV in a human thyroid papillary carcinoma cell line (TPC-1) were examined. When TPC-1 cultures preheated at 40.5 degrees for 48 hr were infected with HCMV and incubated at a supraoptimal temperature (40.5 degrees), the cultures could be maintained for at least 65 days without detection of infectious virus. In contrast, when the infected cultures were incubated at 37 degrees, HCMV persistently infected cultures were established. HCMV was reactivated from the latently infected cultures by decreasing the incubation temperature from 40.5 to 37 degrees, and the cultures subsequently entered into virus persistent infection. Although HCMV-specific polypeptides which comigrate with the immediate early virus polypeptides and nuclear antigens were continuously detectable in the majority (more than 95%) of the cells during the latent period, a detectable level of virus-specified DNA polymerase (one of the early virus proteins) was not induced, suggesting that the blockage of HCMV replication in the latently infected cultures occurs at the early stages of the HCMV replication cycle. Infectious center assay revealed that 0.002 to 0.2% of the cells contain an HCMV genome that can be activated during the latent period. The latently infected cells were susceptible to superinfection with homologous and heterologous strains of HCMV. In persistently infected cultures approximately 38% of the cells were lysed by reaction with HCMV immune serum and complement, whereas complement-mediated immune cytolysis could not be detected in the latently infected cultures. The data presented suggest that a temperature-sensitive cellular function(s) that controls the expression of the HCMV early functions plays an important role in maintenance of the HCMV genome in the latent state and reactivation of HCMV by decreasing the incubation temperature.

Antigens, Viral↗

Spin, oxidation, and ligand states of p-nitrothiophenolatoiron(III) complex of protoporphyrin-IX-dimethylester in the presence of 1-MeIm: magnetic circular dichroism and 1H nuclear magnetic resonance spectroscopic studies.

Complex formation of 5-coordinated iron(III) heme containing thiolate anion (p-nitrothiophenol) with imidazole (1-methylimidazole) showed very interesting features depending on the nature of the solvent and the ratio of the ligand to heme. The complexes formed under different conditions were not only low spin iron(III) complexes with a thiolate anion and an imidazole or with two imidazoles, but also reduced (iron(II] complexes with a thiolate and an imidazole or with two imidazoles. Absorption, magnetic circular dichroism, and 1H NMR spectroscopies could identify the complex formed when they were used concurrently. The dependence of polarity of the solvents used on the resultant chemical species was ascribed to the stability of Fe(III) or Fe(II) complex in the different solvents. The iron(III) complex with a thiolate anion and an imidazole was found to be reduced automatically to the iron(II) complex with a thiolate and an imidazole which exchanged ligand to the iron(II) bisimidazoles in the presence of excess imidazole. This study showed that the ligands of heme are easily exchanged and that the heme iron(III) is automatically reduced in several conditions. Possible significance with respect to biological systems containing a sulfur ligand is discussed.

Circular Dichroism↗

Metabolic pathways of carotenoids in chum salmon Oncorhynchus keta during spawning migration.

1. Based on the contents and individual composition of carotenoids in the muscle, serum and ovaries of chum salmon during spawning migration, the reductive metabolism of astaxanthin to zeaxanthin was presumed to take place in the muscle of both male and female. 2. The metabolic rates of zeaxanthin and 4-keto-zeaxanthin in female serum were much faster than those in male serum.

Animals↗

Inhibitory effect of papaverine on HVJ (Sendai virus) replication in rat glioma C6 cells.

The replication of HVJ (Sendai virus) in C6 rat glial cells was found to be inhibited by treatment of the cells with papaverine, an inhibitor of cAMP phosphodiesterase, but not with cAMP or dibutyryl cAMP. In addition, cyclic GMP which often manifests a reciprocal relationship to cAMP did not counteract the inhibition of HVJ yield by papaverine. Both viral genome replication and transcription were suppressed slightly by treatment of the cells with papaverine. In the cells cultured in the presence of papaverine, the synthesis of viral proteins and their phosphorylation occurred at normal rates. Membrane immunofluorescence and cell surface immunoprecipitation showed that the viral glycoproteins HN and F0 were expressed on the cell surface of the papaverine-treated cells. Moreover, all the viral structural proteins were associated with plasma membrane isolated from the treated cells. These results indicate that papaverine treatment suppresses some part of the process of virus budding at the plasma membrane.

Animals↗

Clinical evaluation and characterization of a unique C3 breakdown factor detected in a patient with acute glomerulonephritis.

A unique factor causing breakdown of the third component of complement (C3) was detected in a patient with acute glomerulonephritis (AGN). The activity was detected by conversion of C3 in a mixture with normal human serum using crossed immunoelectrophoresis. This factor capable of C3 breakdown was not blocked in the presence of ethylenediaminetetraacetic acid (EDTA), heat-pretreatment (56 degrees C, 30 min) und dialysis against 0.1 M phenylmethylsulfonyl fluoride (PMSF). The factor was isolated and characterized by immunochemical and physicochemical techniques. It had beta-mobility on electrophoresis and was separated at a lighter sedimentation coefficient than 7S by sucrose density gradient ultracentrifugation. The activity was absorbed on a DEAE-cellulose column and was eluted under definite conditions of relative salt concentration. These data appear to suggest that this factor is distinct from C3 nephritic factor (C3NeF), immunoglobulins, immune complexes, and other C3 convertases previously described. Moreover, it is clear that the factor was mainly responsible for the C3 activation observed in the patient with acute glomerulonephritis.

Acute Disease↗