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Biomedical subjects

H Hirano

Publications and source records attributed to H Hirano.

At least 343 records · Page 19Linked to original sources

[Distribution of glycoconjugates in gerbil hippocampal neurons--histochemical study with lectins].

Paraffin-embedded sections of gerbil hippocampus were made and stained by use of lectins with different sugar specificities: Concanavalin A (Con A), wheat germ agglutinin (WGA), Ricinus communis agglutinin (RCA), Phaseolus vulgaris agglutinin (PHA-E), peanut agglutinin (PNA), soybean agglutinin (SBA), and Vicia villosa agglutinin (VVA) to investigate the distribution of glycoconjugates in this brain region. Nuclear membranes of all the neurons in the hippocampus were positively stained with Con A and PHA-E, whereas WGA revealed definite staining of cell membranes. Endothelial cells of blood vessels were intensely positive to RCA, suggesting its usefulness as a marker of endothelial cells. With SBA and VVA, a few neurons in Ammon's horn were positively stained, while no positive cells were observed in the dentate gyrus. CA2 and the medial part of the CA1 were also positive bilaterally with SBA and VVA. Some neurons in Ammon's horn and dentate gyrus were selectively stained on the surface of their cell bodies with PNA. The present results show that lectins used distinguish different subpopulations of hippocampal neurons, indicating a new possible classification of hippocampal neurons based on their differences in glycoconjugates.

Animals↗

[A case report on tuberculosis with remittent fever after tonsillectomy in a patient under CAPD].

This report pertains to a case of chronic renal failure with remittent fever after tonsillectomy. The patient was 45-year-old female who had been undergoing continuous ambulatory peritoneal dialysis (CAPD) for five years. She was admitted to our hospital after being diagnosed as having pyrexia with tonsillitis. A tonsillectomy was performed. Although several symptoms and signs, such as fever, positive CRP and accelerated ESR, improved transiently by the seventh postoperative day, remittent fever and cervical lymph node swelling suddenly recurred after the eighth postoperative day. In spite of the antibiotic therapy, the fever continued for two weeks thereafter. A culture to check for acid-fast bacilli was negative, but on epithelioid cell granuloma with a small central abscess was found in the biopsy specimen of the lymph node. INH was prescribed to her. After three days of INH administration, the patient became afebrile. Patients with long-term dialysis are known to be very susceptible to tuberculous diseases. However, to date, there has been no report of tuberculosis being accompanied by a fever after a tonsillectomy. As it might be difficult to make a tubercular diagnosis on such a febrile patient, early antituberculous chemotherapy is recommended for patients with antibiotic-refractory fever.

Female↗

The influence of experimentally produced oligohydramnios on lung growth and pulmonary surfactant content in fetal rabbits.

To study the effect of oligohydramnios on lung growth and biochemical lung development in fetal rabbits, amniotic fluid was drained through a tube inserted into the maternal peritoneal cavity on the 23 day of gestation. Littermate fetuses without an amniotic shunt were used as controls. The fetuses were delivered abdominally on the 28 day of gestation. In a total of 8 pregnant does, 17 fetuses underwent amniotic shunting and 22 fetuses were used as controls. The amniotic shunt produced a significant reduction in the amniotic fluid volume. There were no differences in the wet weights of the fetal body, liver or brain between the two groups. However, the amniotic shunt significantly decreased the wet weight of the fetal lung, fetal lung wet weight/body weight ratio, and protein concentration per lung as compared to the control fetuses. In the fetal liver and brain tissues, no changes were found in the concentrations of total phospholipids, phosphatidylcholine (PC) or disaturated phosphatidylcholine (DSPC, the main component of lung surfactant) per g of wet tissue and per mg of protein. However, the lungs of the fetuses with amniotic shunts contained significantly more PC and DSPC, and the L/S ratio was higher than in the control fetuses. These results suggest that the oligohydramnios produced by an amniotic shunt causes pulmonary hypoplasia, but raises the pulmonary surfactant content of fetal rabbit lung.

Animals↗

Induced bone development in transplants of fresh human pseudomalignant heterotopic ossification tissue in athymic nude mice.

Biopsy specimens of mature trabecular bone from tumors of two typical cases of pseudomalignant heterotopic ossification (PHO; myositis ossificans circumscripta) were transplanted into athymic nude mice. Specimens of normal metaphyseal bone in adjacent areas were also transplanted in the contralateral hindquarter muscles for controls. By seven days, control bone transplants were necrotic and enveloped in granulation tissues whereas PHO transplants were surrounded by proliferating connective tissue derived from the host-bed muscle tissues. In the intervals from 14 and 21 days, the PHO transplants showed vascular and hypertrophied connective-tissue proliferation with appositional deposits of new bone. The microscopic features of the new bone suggested that it was of mouse host-bed origin because the new bone from Case 1, a female, showed nuclei without sex chromatin. The quantities of new bone were measured by correlated histomorphometric and computer image analysis of microradiographs. Little or no cartilage development was noted at any stage. The normal control trabecular bone tissue was slowly resorbed by macrophages and mononuclear phagocytes; multinucleated giant cells were few and relatively small. Except in one questionably small area of one transplant, nontumorous undemineralized normal human bone failed to induce bone formation within the 28-day period of observation. The quantity of new bone formed in athymic mice in response to implants of PHO, estimated from equivalent quantities of bone developed from implants of lyophilized matrix-free bone morphogenetic protein (BMP) was about 1 microgram/g of wet PHO tumor weight. Although circumstantial evidence implicates BMP in the pathogenesis of heterotopic ossification, more research should be directed toward regional and systemic factors inhibiting bone development in normal and abnormal conditions, including malignant bone tumors.

Adult↗

[Step section of the TBLB specimens in the diagnosis of sarcoidosis].

The authors analyzed a total of 132 cases clinically diagnosed as sarcoidosis from 1979 to 1990. Routine TBLB sections revealed sarcoid granulomas in 57 cases (Stage 0; 5%, Stage 1; 40%, Stage 2; 57%). In 75 cases in whom routine sections did not show granuloma, we prepared additional sections from paraffin-embedded blocks as step sections. When examined with step sections in addition to routine specimens, the detection rate of sarcoid granuloma increased to 47% in Stage 1 and 82% in Stage 2. We concluded that making step sections of TBLB specimens is a useful diagnostic method in cases suspected of sarcoidosis, especially in Stage 2 cases.

Adolescent↗

Ultracytochemical localization of the erythrocyte/HepG2-type glucose transporter (GLUT1) in the ciliary body and iris of the rat eye.

Aqueous humor, with its unique low concentration of proteins, is produced by the ciliary body and isolated by the blood-aqueous barrier from the body fluid. Glucose in aqueous humor is a major source of nutrients for lens and corneal cells, and is maintained near the plasma level, suggesting a specific glucose transport mechanism in the blood-aqueous barrier. Using antibodies against erythrocyte/HepG2-type glucose transporter (GLUT1), one isoform of the facilitated diffusion glucose transporters, the authors found immunocytochemically that GLUT1 localizes in the epithelial cells of ciliary body and iris. GLUT1 is also found in the endothelial cells of blood vessels in the iris, whereas no labeling is seen in the blood vessels in the ciliary body. In the ciliary body epithelium, the plasma membranes of both the pigmented epithelial (PE) and nonpigmented epithelial (NPE) cells are positive for GLUT1. By the colloidal gold particle counting, the basal infoldings of PE cells show approximately two-fold denser labeling than those of NPE cells. Since PE and NPE cells make up a functional syncytium with numerous gap junctions, the authors suggest that glucose transport in the ciliary body occurs in this manner: glucose diffuses out from blood vessels through the pores of fenestrated endothelial cells, is transported into PE cells by GLUT1 in their plasma membrane, enters NPE cells through gap junctions connecting PE and NPE cells, and is finally transported into the aqueous humor by GLUT1 of NPE cells. The higher density of GLUT1 in PE cells may account for the consumption of glucose by PE and NPE cells in addition to the transepithelial transport.

Animals↗

Erythrocyte/HepG2-type glucose transporter is concentrated in cells of blood-tissue barriers.

In search of possible diverse roles of glucose transporters (GT's), we examined whether any GT's are present in blood-tissue barriers where selective flow of glucose from blood to tissue cells is critically important. We found in rat that the erythrocyte/HepG2-type GT is localized in all the limiting plasma membranes known to serve as blood-tissue barriers, whether the barriers are endothelial type (brain, iris, inner retina, peripheral nerve) or epithelial type (choroid plexus, ciliary body, outer retina, peripheral nerve, placenta), except for plasma membranes in testis and thymus where no appreciable amount of the GT was found. The erythrocyte/HepG2-type GT may play a vital role for the entry of glucose into these firmly guarded tissues.

Actins↗

Translocation of protein kinase C to membranes induced by TNF does not cause the inhibition of EGF binding to human wish cells.

Tumor necrosis factor (TNF) caused an inhibition of 125I-labeled epidermal growth factor [( 125I]EGF) binding to its receptors of human amniotic (WISH) cells at 5 min after addition of TNF, which reached a maximal level (60-70% reduction) after 15-30 min and declined thereafter. TNF also induced a translocation of protein kinase C activity from the cytosol to the membrane, which peaked at 45-60 min after addition of TNF and almost returned to basal level at 120 min. Furthermore, prolonged incubation of WISH cells with 12-O-tetradecanoylphorbol 13 acetate (TPA) diminished the TPA effect on the inhibition of EGF binding to the cells due to the desensitization of protein kinase C; however, TNF still reduced the EGF binding to the cells pretreated with TPA for a long time. These results indicate that although TNF causes the translocation of protein kinase C to the membrane, activation of protein kinase C is not required for TNF to induce a decrease in EGF binding to the cells.

Amnion↗

Microsequencing of proteins electrotransferred onto immobilizing matrices from polyacrylamide gel electrophoresis: application to an insoluble protein.

Semidry electroblotting is convenient and allows a rapid and efficient protein transfer from one- or two-dimensional polyacrylamide gels onto sequencer stable supports for protein microsequence analysis in a gas-phase sequencer. Using this technique, we determined the amino acid sequences of the basic 7S globulin (Bg), an insoluble protein present in soybean seeds. Based on sequence determination, the cDNA-encoding Bg could be easily cloned and characterized.

Amino Acid Sequence↗

Localization of Forssman glycolipid and GM1 ganglioside intracellularly and on the surface of germ cells during fetal testicular and ovarian development of mice.

Changes in the expression pattern and intracellular localization of Forssman glycolipid (FA) and GM1 ganglioside (GM1) in fetal mouse gonads were examined during germ cell differentiation by immunofluorescence microscopy and immunoelectron microscopy. In male germ cells from the 12th to 14th day p.c., anti-FA binding was localized in granular structures aggregated on one side of the cytoplasm and/or in the plasma membrane. On day 16 p.c., some germ cells still showed patch-like positive reactions in the plasma membrane, but by day 18 p.c., positive reactions for FA had completely disappeared. The female germ cells showed granular bindings of anti-FA scattered throughout their cytoplasm during the 13th to 16th day p.c., although the positive reactions in female germ cells on day 12 p.c. tended to be found in one side of cytoplasm and/or plasma membrane similar to those in male germ cells from 12th to 14th day p.c. On day 18 p.c., positive reactions remained in the plasma membrane of some germ cells, but these positive reactions disappeared before birth. Immunoelectron microscopic observation showed that the sites of anti-FA bindings were equivalent to the "small dense bodies" (SDB) and the Golgi lamellae both in male and female germ cells. On the other hand, GM1 was not detected in male germ cells at any time during fetal testicular development, whereas an anti-GM1 reaction was detected in the plasma membrane of female germ cells from the 16th to 18th day p.c. (oocytes in the first meiotic prophase).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Simultaneous activation of heat shock protein (hsp 70) and nucleolin genes during in vivo and in vitro prereplicative stages of rat hepatocytes.

Rapidly growing cells usually have high levels of ribosome biogenesis. The sequential expression of protooncogenes during the transition of quiescent hepatocytes to the replicative stage was assumed to be followed by activation of cellular genes related to cell growth such as ribosome biosynthesis. First, the expression of major nucleolar protein (nucleolin or C23) and major heat-shock protein (hsp 70) genes was examined during rat liver regeneration. hsp 70 may function in cell growth and has a characteristic nucleolar location after heat shock. Both nucleolin and hsp 70 mRNA began to increase simultaneously after peaks of c-fos and c-myc, showed a peak 6 h after partial hepatectomy, and declined to the control levels around 20 h. That is, the peaks of nucleolin and hsp 70 mRNA precede the peak of ribosome formation (12-20 h) and DNA replication (24 h). Second, the behavior of nucleolin and hsp 70 mRNA was examined in primary cultured hepatocytes during their G0-G1 transition. Although the amounts of c-myc mRNA reached a plateau around 20 h after the initiation of culture and remained at these levels, DNA synthesis has never been found to start without the addition of EGF and insulin to this system. Both nucleolin and hsp 70 mRNA began to increase at around 20 h (prereplicative stage) and simultaneously decreased in inverse proportion to DNA synthesis induced by these growth factors. Thus, it is possible that the simultaneous enhancement of nucleolin and hsp 70 genes as described above is not merely coincidental, but is important biologically during the transition of quiescent hepatocytes to proliferative cells.

Animals↗

Calcium-induced aggregation of the urea-deaggregated human lens insoluble protein.

Calcium-induced aggregation of the urea-deaggregated insoluble protein (US-fraction) derived from human lens was investigated by use of a simple light scattering method. The initial rate of aggregation was shown to increase with progress of senescence of the lens, and to be much higher in the nucleus as compared to the cortex. The aggregation was found to be more active in the US-fraction from cataractous lens than in that of normal lens at same age. A calcium sensitive polypeptide (CSP) was isolated from the US-fraction from cataractous human lenses by gel filtration followed by DEAE-cellulose column chromatography in the presence of 7 M urea. The molecular size of the CSP in the dissociated state was assessed to be 4.2 kDa by urea-SDS-PAGE, and the size in the associated state to be 450 kDa by thin layer gel filtration. With addition of calcium ion to the latter, it aggregates rapidly to form much larger aggregates and eventually precipitates out. The aggregation rate of the CSP is shown to increase exponentially with an increase in the concentration of both CSP and calcium ion, indicating the involvement of complicated mechanisms in the aggregation process. Effects of divalent ions other than calcium ion on the aggregation process were also examined. It is revealed that the effect of manganese ion is little less than that of calcium ion, whereas magnesium and strontium ions are much less effective as compared to the former two ions.

Calcium Chloride↗

Differences in lectin binding patterns of normal endometrium and endometrial adenocarcinoma, with special reference to staining with Ulex europeus agglutinin 1 and peanut agglutinin.

Differences in glycoconjugate composition between proliferative endometrium and endometrial adenocarcinoma were investigated by histochemical techniques using seven different lectins as probes. For light microscopy, the avidin-biotin-peroxidase complex (ABC) method was used. Concanavalin A (Con A), wheat germ agglutinin (WGA), and Ricinus communis agglutinin (RCA) stained almost all glandular cells in both proliferative (normal) and malignant endometria. Ulex europeus agglutinin 1 (UEA-1) strongly stained cancer cells, especially well-differentiated adenocarcinoma, but it scarcely stained normal endometrium. Peanut agglutinin (PNA) binding sites were observed only along the apical surface of normal endometrial glands, while the cytoplasm of endometrial adenocarcinoma cells was often positive for PNA. Soybean agglutinin (SBA) faintly reacted with proliferative endometrium and occasionally with malignant cells. Dolichos biflorus agglutinin (DBA) slightly stained proliferative and malignant endometria. By electron microscopic examinations using horseradish peroxidase (HRP)-labeled lectins, we observed that UEA-1, PNA, and Con A stained the Golgi membranes and plasma membrane of carcinoma cells. In addition, the Con A reaction was positive in the endoplasmic reticulum and nuclear envelope. These results revealed the differences in oligosaccharide chains between normal and malignant endometria, suggesting that UEA-1 and PNA, in particular, may be useful indicators of malignancy of the endometrium.

Adenocarcinoma↗

Isolation and structure of cecropins, inducible antibacterial peptides, from the silkworm, Bombyx mori.

1. Cecropins, inducible antibacterial peptides, were purified by simple two step chromatography from immunized larval hemolymph of the silkworm, Bombyx mori. 2. The cecropins were further separated into four major and two minor forms by reverse-phase HPLC. 3. The four major cecropins were sequenced and divided into two types, A and B, whose structures were quite similar to cecropins A and B of Hyalophora cecropia. 4. Three of them contained an unusual amino acid, delta-hydroxylysine. 5. No remarkable difference in antibacterial activity against E. coli was detected among these cecropins.

Amino Acid Sequence↗

Ca2+ entry blockers, force staircase and the onset of the positive inotropic action of cardiotonic steroids in isolated cardiac muscle.

1. Effects of alterations of Ca2+ fluxes on the force staircase phenomenon and on the positive inotropic action of strophanthidin or ouabain were examined in left atrial muscle preparations isolated from rabbit or rat heart. 2. The organic Ca2+ entry blockers converted the positive force staircase observed in rabbit heart to the negative staircase; however, Ni2+, Co2+ or a reduction of the extracellular Ca2+ concentration which reduces Ca2+ influx via the Na+/Ca2+ exchange mechanism as well as via the Ca2+ channels, failed to convert the force staircase. 3. All of these interventions or ryanodine, which inhibits Ca2+ release from the sarcoplasmic reticulum, delayed the onset of the positive inotropic effect of strophanthidin or ouabain without reducing the peak inotropic effect. 4. These results indicate that either Ca2+ antagonists reduce the Na+ influx in addition to reducing Ca2+ influx, or Ca2+ influx per se and not the Na+ influx is important for the staircase phenomenon, and that intracellular Ca2+ accumulation plays an important role in the early phase of the positive inotropic effect of the cardiotonic steroids.

Animals↗

Cloning and characterization of a gene encoding a new thermostable hemolysin from Vibrio parahaemolyticus.

A new thermostable hemolysin (delta-VPH) gene was cloned from a Kanagawa-negative Vibrio parahaemolyticus strain into vector pBR322 in Escherichia coli K12. The nucleotide and amino acid sequences had no homology with those of the thermostable direct hemolysin (TDH) which causes the Kanagawa phenomenon, and of the thermolabile hemolysin (TLH) of V. parahaemolyticus. The gene was present in all V. parahaemolyticus strains tested and also in one strain of V. damsela.

Amino Acid Sequence↗