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Biomedical subjects

H Hirano

Publications and source records attributed to H Hirano.

At least 217 records · Page 12Linked to original sources
PubMed1995

Production of waxy (amylose-free) wheats.

The Waxy (Wx) protein has been identified as granule-bound starch synthase (GBSS; EC 24.1.21), which is involved in amylose synthesis in plants. Although common wheat (Triticum aestivum L.) has three Wx proteins, "partial waxy mutants" lacking one or two of the three proteins have been found. Using such partial waxy mutants, tetra- and hexaploid waxy mutants with endosperms that are stained red-brown by iodine were produced. Both mutants showed loss of Wx protein and amylose. This is the first demonstration of genetic modification of wheat starch.

Electrophoresis, Polyacrylamide Gel↗
PubMed1995

Cloning, sequencing and expression of a novel cDNA encoding human vacuolar ATPase (14-kDa subunit).

A cDNA encoding the 14-kDa subunit of vacuolar ATPase was cloned from human fetal brain. The sequence was composed of 680 nucleotides containing an open reading frame of 357 nucleotides. The deduced peptide sequence consisted of 119 amino acid residues with a calculated molecular weight of 13,369 Da and a pI of 5.19. Overall, this amino-acid sequence was respectively 69% and 70% identical to those of Manduca sexta and Drosophila melanogaster 14-kDa subunits, although the two representatives of Class Insecta were remarkably similar to one another (91% identity). Three regions in particular (the N-terminal, amino acids 5-36; the middle, residues 58-68; and the C-terminal, residues 88-118) were highly conserved. Hence, we think that the 14-kDa subunits have evolved from the same ancestral gene, and that the three conserved regions are important for the structure and function of vacuolar ATPase. A single 0.8-kb band was detected in various human tissues by Northern blot analysis. Since the human 14-kDa subunit is expressed ubiquitously, it might be a housekeeping protein. A separate transcript found in the cDNA library lacked a 6-bp segment in the 5' non-coding region (nucleotides -40 to -35) and also carried a 23C to T (8Thr to Ile) point mutation in the coding region; these minor differences likely reflected normal polymorphism.

Amino Acid Sequence↗
PubMed1995

Purification and characterization of Rab GDI beta from rat brain.

A new Rab GDI was purified from the synaptic soluble fraction of rat brain by several column chromatographies as a protein that inhibited the dissociation of [3H]GDP from Rab3A but was not recognized by an anti-Rab GDI alpha antibody. The partial amino acid sequence analysis revealed that it was identical with rat Rab GDI beta. Purified Rab GDI beta showed the kinetic properties similar to those of Rab GDI alpha, including the inhibitory effect on the dissociation of GDP from Rab3A, the substrate specificity, the requirement of the post-translational lipid modifications of Rab3A, the stoichiometric interaction with the GDP-bound form of Rab3A, the inhibitory effect on the binding of Rab3A to erythrocyte ghosts, and the stimulatory effect on the dissociation of Rab3A from the membrane.

Amino Acid Sequence↗
PubMed1995

Immunocytochemical localization of the protein reactive to human beta-1, 4-galactosyltransferase antibodies during chick embryonic skin differentiation.

BACKGROUND: beta-1, 4-Galactosyltransferase (GalTase) transfers galactose from UDP-galactose to terminal N-acetylglucosamine in glycoconjugates and is located both in the Golgi apparatus and in the plasma membrane. The cell surface GalTase is thought to be involved in cell-to-cell recognition and cell-to-extracellular matrix interaction. METHODS: By the use of specific monoclonal antibodies against human GalTase, changes in cell surface localization of the protein reactive to the antibodies in chick embryonic skin during its differentiation in vivo and in vitro were detected immunohistochemically at both light- and electron microscopic levels. The distribution of glycoconjugates having terminal N-acetylglucosamine residues was detected by staining with succinylated wheat germ agglutinin (s-WGA). RESULTS: Under the light microscope, intense immunostaining was observed in the keratinized epidermis, particularly in the intermediate layer. Marked changes in the localization of the staining were observed in vitamin A-induced mucus-secreting skin, in which keratinization was suppressed. The localization of the immunostaining was in parallel with that of glycoconjugates having terminal N-acetylglucosamine residues. Immunoelectron microscopically the immunostaining was located on the cell surface and in the intercellular space of the desmosomes in the intermediate cells of the keratinized epidermis. However, the staining was not present on the cell surface but was detected on the limiting membrane of the mucous granules, in the mucous metaplastic epidermis. In contrast, the staining was always found in the Golgi apparatus in all of the cells. CONCLUSIONS: These results suggest that the protein reactive to human GalTase antibody may be involved in chick epidermal differentiation.

Animals↗
PubMed1995

EDA-containing fibronectin is synthesized from rheumatoid synovial fibroblast-like cells.

OBJECTIVE: To identify the cells that synthesize EDA-containing fibronectin (FN) and examine the role of EDA+FN in the pathogenesis of rheumatoid joint lesions. METHODS: Localization of EDA+FN and c-Fos protein in rheumatoid joints was studied immunohistochemically by utilizing antibodies for EDA+FN and c-Fos. Expression of EDA+FN was studied by immunoelectron microscopy and in situ hybridization. The amount of EDA+FN was measured by enzyme-linked immunosorbent assay. RESULTS: EDA+FN was specifically localized in the synovial lining layer of synovium with active rheumatoid arthritis (RA) (n = 17), but not in that with osteoarthritis (n = 4) or with inactive fibrous RA (n = 2). EDA+FN messenger RNA was localized in the synovial lining layer. EDA+FN was immunoelectron microscopically localized in the synovial lining fibroblast-like (type B) cells. EDA+FN was also detected at the cartilage-pannus junction and on the surface of RA cartilage. Double staining showed that EDA+FN colocalized with c-Fos protein in the rheumatoid synovial lining layer. Quantification of EDA+FN showed that it was highly concentrated in rheumatoid synovial fluids. CONCLUSION: EDA+FN is synthesized by the synovial lining fibroblast-like (type B) cells in situ in rheumatoid synovium, and appears to be expressed in association with activated or transformed states of synovium.

Animals↗
PubMed1995

Thioredoxin h is one of the major proteins in rice phloem sap.

Sieve tubes play important roles in the transfer of nutrients as well as signals. Hundreds of proteins were found in pure phloem sap collected from rice (Oryza sativa L. cv. Kantou) plants through the cut ends of insect stylets. These proteins may be involved in nutrient transfer and signal transduction. To characterize the nature of these proteins, the partial amino-acid sequence of a 13-kDa protein, named RPP13-1, that was abundant in the pure phleom sap was determined. A cDNA clone of 687 bp, containing an open reading frame of 122 amino acids, was isolated using corresponding oligonucleotides as a probe. The deduced amino-acid sequence was very similar to that of the ubiquitous thiol redox protein, thioredoxin. The consensus sequences of thioredoxins are highly conserved. No putative signal peptide was identified. Antiserum against wheat thioredoxin h cross-reacted with RPP13-1 in the phloem sap of rice plants. RPP13-1 produced in Escherichia coli was reactive to antiserum against wheat thioredoxin h. Both E. coli-produced RPP13-1 and the phloem sap proteins catalyzed the reduction of the disulfide bonds of insulin in the presence of dithiothreitol. These results indicate that an active thioredoxin is a major protein translocating in rice sieve tubes.

Amino Acid Sequence↗
PubMed1995

Changes in expression of two endogenous beta-galactoside-binding isolectins in the dermis of chick embryonic skin during development in ovo and in vitro.

In order to elucidate the roles of metal-independent animal lectins, we systematically investigated changes in expression of 2 kinds of beta-galactoside-binding isolectins (MW 14 and 16 kDa) in the dermis of chick embryonic tarsometatarsal skin during the course of development. These lectins were immunohistochemically located at different stages of development both in ovo and in vitro by light and electron microscopy. Light-microscopic observation showed that while positive staining for the 14-kDa lectin was weak at days 8 and 10 it became intense after day 13. In contrast, staining for the 16-kDa lectin was intense at days 8, 10, and 13, but it became weak after day 17 when keratinization of the epidermis was completed. Immuno-electron-microscopic observation revealed that both the 14 and 16-kDa lectins were located on the basement membrane, in the extracellular matrix, and in both the cytoplasm and the nucleus of dermal fibroblasts. Distribution of the 2 isolectins was also examined in cultured skin explants in vitro. The results were almost the same as those obtained in ovo when the skin explant was keratinized in the presence of hydrocortisone. However, in the skin explant where keratinization was prevented and mucous metaplasia was induced by the addition of vitamin A, the distribution of the 14-kDa lectin in the epidermis was significantly affected. These results indicate that (1) the expression of the 2 isolectins is differently regulated in both the dermis and epidermis, (2) the 16-kDa lectin is involved in the early stage of the formation of the dermis and the basement membrane and is replaced by the 14-kDa lectin as keratinization of the epidermis occurs, and (3) the expression of the 2 isolectins in the dermis is not significantly affected by the induction of mucous metaplasia, in contrast to their drastic changes in the epidermis.

Animals↗
PubMed1995

Expression of the endogenous 14-kDa beta-galactoside-binding lectin galectin in normal human skin.

The localization of an endogenous 14-kDa beta-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the beta-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.

Base Sequence↗
PubMed1995

Peanut agglutinin binding to human prolactin-producing pituitary adenomas.

Peanut agglutinin (PNA)-binding sites in human prolactin (PRL)-producing pituitary adenomas were examined by light and electron microscopy together with immunoblot analysis. At the light microscopic level, the majority of the PRL-producing adenoma cells stained positively for PNA in 15 of 20 cases. PNA binding observed in the cytoplasm had a granular appearance. PRL-producing cells adjacent to the adenoma tissue showed negative PNA staining. In normal pituitary glands, the PRL-positive glandular cells were negative for PNA staining. By electron microscopy, reaction products showing PNA-binding sites were detected in some of the secretory granules. Immunoblotting analysis revealed that the PRL bands corresponded to PNA-stained ones with the exception of the main 23-kDa band. PNA-binding sites have some relation to the secretory granules containing glycosylated forms of PRL. These observations suggest that PNA staining can be used as a valuable method to analyze human PRL-producing pituitary adenomas.

Adenoma↗
PubMed1995

Normal variations of the temporal bone on high-resolution CT: their incidence and clinical significance.

We evaluated normal variations of the temporal bone on high-resolution computed tomograms (HR-CTs) and investigated their incidence. HR-CTs of the temporal bones of 325 patients were retrospectively reviewed. Six groups of variants, which were considered important for presurgical planning, were evaluated on HR-CTs. These included: (1) an incomplete bony covering of a high-positioned jugular bulb; (2) severe asymmetry of the jugular foramen; (3) an anteriorly located sigmoid sinus; (4) a deep sinus tympani; (5) a large internal auditory canal; and (6) a large cochlear aqueduct. The frequency of the variations were as follows: (1) 2.4%; (2) 4.0%; (3) 1.6%; (4) 5.9%; (5) 2.3%; and (6) 3.0%. Bilateral involvement with variation (4), (5) and (6) was frequently seen. Normal anatomical variations of the temporal bone are therefore not rare and awareness of the possible variants is necessary before surgery of the inner ear, middle ear and posterior fossa.

Adolescent↗
PubMed1995

Serotonergic regulation of acetylcholine release in rat frontal cortex.

The extent to which serotonin regulates the activity of cortically projecting cholinergic neurons was studied using in vivo microdialysis to monitor interstitial concentrations of acetylcholine in the frontal cortex of freely moving rats. Systemic administration of the serotonin release-inducing agent fenfluramine (3 or 10 mg/kg, i.p.) increased acetylcholine release by 110-130%. The fenfluramine-induced increase in acetylcholine release was significantly attenuated by pretreatment with the selective serotonin uptake inhibitor fluoxetine (10 mg/kg, i.p.). Pretreatment with the selective dopamine D1 receptor antagonist SCH-23390 (0.3 mg/kg, s.c.) failed to prevent the fenfluramine-induced increase in acetylcholine release. In contrast, the serotonin 5-HT2A receptor antagonist ketanserin (5 mg/kg, i.p.) blocked fenfluramine-induced increases in acetylcholine release. In contrast to previous studies that have concluded that serotonin has inhibitory actions on cortical acetylcholine release, the present results indicate that fenfluramine increases cortical acetylcholine release in vivo by its ability to enhance serotonin transmission and that serotonin produces these effects at least in part via actions at serotonin 5-HT2A receptors.

Acetylcholine↗
PubMed1995

Cloning and nucleotide sequence of the gene encoding dimethyl sulfoxide reductase from Rhodobacter sphaeroides f. sp. denitrificans.

The gene encoding dimethyl sulfoxide (DMSO) reductase, which contains a molybdenum cofactor, of the phototrophic bacterium Rhodobacter sphaeroides f. sp. denitrificans was isolated using an oligonucleotide probe, which was synthesized based on a internal amino acid sequence of the purified enzyme. The DMSO reductase gene coded for 822 amino acids (2466 base pairs, M(r) = 89,206) as a precursor form having a signal peptide of 42 amino acids. The deduced amino acid sequence had high homology with those of some enzymes containing a molybdenum cofactor: trimethyl amine N-oxide reductase (48%), biotin sulfoxide reductase (44%), and DMSO reductase (29%) of Escherichia coli.

Amino Acid Sequence↗
PubMed1995

[A case of renovascular hypertension with nephrotic syndrome].

A 68-year-old male patient with renovascular hypertension (RVHT) and nephrotic syndrome (NS) is described. He was admitted to our hospital for detailed investigation of severe hypertension and massive proteinuria. After admission, a diagnosis of RVHT with a right non-functional kidney and NS was made. Nephrectomy and contralateral renal biopsy were performed for refractory hypertension and detailed investigation of the NS, respectively. The renal biopsy showed focal segmental glomerulosclerosis (FGS) in the left kidney, whereas the nephrectomised kidney exhibited only ischemic change. After the operation, his blood pressure became stable without anti-hypertensive agents, but proteinuria remained in the nephrotic range. Six months later, proteinuria had disappeared and his renal function was stable. These findings suggest that NS and FGS might have resulted from an activated renin-angiotensin-axis and that the prolonged NS was due to severe glomerular injury. Although there have been many reports describing the relationship between RVHT and FGS in an experimental environment, this relationship is very rare in clinical cases. Therefore we present this case to increase understanding of the cause of FGS.

Aged↗
PubMed1995

[Clinical application of digital TV tomosynthesis to disorders of the wrist and their post-operative evaluation].

The clinical utility of digital TV tomosynthesis was assessed in 13 patients with chronic disorders of the wrist and the first carpometacarpal joints, and a post-operative evaluation was carried out. The disorders included nonunion of the scaphoid in five patients, Kienböck's disease in four, subluxation of the distal radioulnar joint in two, and osteoarthrosis of the first carpometacarpal joint in two. The total time needed to obtain two sets of tomographs of a wrist by digital TV tomosynthesis was about 10 minutes, or approximately one third of the time required with conventional methods. The total radiation exposure in obtaining all tomographs of one direction with digital TV tomosynthesis was less than that in obtaining only one tomograph with conventional methods. Digital TV tomosynthetic images were judged superior to conventional tomographs by four orthopedic surgeons and four radiologists, because tomographic blur was reduced in part by a spatial frequency filter processing. Clinically, digital TV tomosynthesis was very helpful in evaluating nonunion of the scaphoid and Kienböck's disease, especially in their post-operative conditions with metallic fixative devices. Digital TV tomosynthesis was found to be a fast, feasible and inexpensive method with low radiation exposure for evaluating disorders of the wrist.

Adult↗
PubMed1995

Modulation of B-cell abnormalities in lupus-prone (NZB x NZW)F1 mice by normal bone marrow-derived B-lineage cells.

(NZB x NZW)F1(NZB/WF1) mice spontaneously develop an autoimmune disease characterized by abnormality of haemopoietic stem cells. The present study examined a possible regulatory cell interaction between NZB/WF1 and normal bone marrow cells using radiation-induced chimeras. We demonstrated that the ability of NZB/WF1 bone marrow cells to transfer the typical disease with hypergammaglobulinemia including autoantibodies into lethally irradiated normal recipients was prevented by cotransfer of bone marrow from normal CBA/J mice but not from xid CBA/N mice carrying a selective defect in B-cell function. Flow cytometric analysis revealed that the generation of NZB/WF1 cells was reduced in the mixed chimeras given CBA/J but not CBA/N bone marrow cells. Interestingly, radiation chimeras reconstituted with a mixture of NZB/WF1 bone marrow and CBA/J splenic B cells did not show elevation of serum immunoglobulin levels, although most of the spleen cells were dominated by NZB/WF1 cells. On the other hand, NZB/WF1 B cells maturated in vivo in the presence of CBA/J bone marrow or splenic B cells lost the hyper-responsiveness to lipopolysaccharide (LPS) in the autoantibody production in vitro. These results suggest that radiosensitive normal B-lineage cells have the regulatory activity to ameliorate the hypergammaglobulinemia of NZB/WF1 mice by reducing the generation of NZB/WF1 B cells and/or by correcting their hyper-responsiveness, and that NZB/WF1 mice may have a defect(s) in the regulatory cell function. In addition, CBA/J splenic B cells were shown to modulate the B-cell abnormality even when injected into non-irradiated NZB/WF1 mice manifesting autoimmunity.

Animals↗
PubMed1995

Immunohistochemical localization of Na(+)-dependent glucose transporter in the rat digestive tract.

Glucose is actively absorbed in the intestine by the action of the Na(+)-dependent glucose transporter. Using an antibody against the rabbit intestinal Na(+)-dependent glucose transporter (SGLT1), we examined the localization of SGLT1 immunohistochemically along the rat digestive tract (oesophagus, stomach, duodenum, jejunum, ileum, colon and rectum). SGLT1 was detected in the small intestine (duodenum, jejunum and ileum), but not in the oesophagus, stomach, colon or rectum. SGLT1 was localized at the brush border of the absorptive epithelium cells in the small intestine. Electron microscopical examination showed that SGLT1 was localized at the apical plasma membrane of the absorptive epithelial cells. SGLT1 was not detected at the basolateral plasma membrane. Along the crypt-villus axis, all the absorptive epithelial cells in the villus were positive for SGLT1, whose amount increased from the bottom of the villus to its tip. On the other hand, cells in the crypts exhibited little or no staining for SGLT1. Goblet cells scattered throughout the intestinal epithelium were negative for SGLT1. These observations show that SGLT1 is specific to the apical plasma membrane of differentiated absorptive epithelial cells in the small intestine, and suggest that active uptake of glucose occur mainly in the absorptive epithelial cells in the small intestine.

Animals↗
PubMed1995

[Local chemotherapy for malignant brain tumors using methotrexate-containing fibrin glue].

Fibrin glue (FG) is an agent developed for achieving hemostasis and the adhesion of living tissue during surgical operations. Incorporation of a drug into FG may be expected to have a sustained local release. In the present study, methotrexate (MTX) included in FG (FG-MTX) was used. The release of MTX into human plasma and cerebrospinal fluid was studied by in vitro study to confirm the sustained release effect of this preparation, by in vivo study, in which the antitumor effect of FG-MTX was assessed in rats bearing 9L-gliosarcoma subcutaneously; and clinically, FG-MTX therapy was attempted in patients with malignant brain tumors. The in vitro study showed that MTX levels rapidly decreased over 1 to 3 days, but was still detected on days 7 and 14. The results showed the sustained release effect of MTX. The in vivo study showed that in the FG-MTX group, all tumors began to decrease soon after administration and disappeared in four out of five animals (80%) on about day 10. In the clinical study, sustained release for more than one week was found, and tumor decrease occurred in the case of a malignant brain tumor. Thus, FG-MTX appears to provide an effective local chemotherapy.

Animals↗