Biomedical subjects

H Hirano

Publications and source records attributed to H Hirano.

At least 325 records · Page 18Linked to original sources

PubMed1991

Unusual biochemical development of genetically seizure-susceptible El mice.

Looking for the 'epilepsy gene', we used ddY derived, genetically seizure-susceptible El mice. To find biochemical abnormalities, we examined the amino acid metabolism and gene activity, including poly(A)+ RNA and sodium channel mRNA expressions, in the developmental growth of El mice. At the early postnatal stage, abnormalities in amino acid metabolism were aberrant free amino acid fluctuations. Almost all free amino acids in the liver of newborn El mice showed considerably lower levels than did ddY mice. Among those amino acids, Asp, Glu and Tyr were extremely low, but rapidly recovered to the ddY level within a week. During the successive growth period, we observed no significant difference in hepatic amino acid levels between El and ddY mice. No such drastic changes were noted in the amino acid levels in the brains of ddY and El mice; only the Gly level was greater in El mice than in ddY mice on the day of birth. Rotatory stimulation which evokes convulsions in El mice but not in ddY mice was applied to adult mice and changes in the amino acid level were assessed. The level of Glu and Tyr in seizure-induced El mice was approximately twice that noted in the liver and brain of El mice, which did not experience seizures. It was also somewhat increased in ddY mice subjected to rotational stress which did not induce seizures in that strain. Gene activity that expresses poly(A)+ RNAs, including sodium channel mRNA, was determined by Northern blot analysis, which reveals unscheduled mRNA synthesis by the appearance of an extra band approximately 3 kb in size.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

PubMed1991

Plant basic 7 S globulin-like proteins have insulin and insulin-like growth factor binding activity.

Basic 7 S globulin (Bg) is a cysteine-rich glycoprotein present in soybean seeds. Mature Bg is composed of high- and low-kDa subunits linked by disulfide bonding. A ligand blotting experiment using [125I]insulin and [125I]insulin-like growth factor-I and -II showed that Bg subunits are able to bind not only to insulin but to insulin-like growth factors-I and -II. Bg-like proteins from other legume species cross-reacted with anti-Bg antibody also bind to insulin and insulin-like growth factors. Bg-like protein in carrot cells was found to have insulin binding activity. Bg-like proteins may be involved in an insulin-like regulatory mechanism in many plant species.

Glycoproteins↗

PubMed1991

Adhesion of immature thymocytes to thymic stromal cells through fibronectin molecules and its significance for the induction of thymocyte differentiation.

Only 10-15% of unseparated thymocytes adhered to culture plates precoated with fibronectin (FN), but 60-70% of the CD4-8- (double-negative) thymocyte population did. This population bound to FN but not to collagen, laminin, or vitronectin. Its binding to FN was inhibited by anti-FN antibody or a mixture of synthetic peptides corresponding to two different sites of FN, termed the V10 sequence and the RGDS (Arg-Gly-Asp-Ser) sequence, which interact, respectively, with the VLA-4 and VLA-5 FN receptors expressed on T-lineage cells. CD4-8- thymocytes also adhered to a monolayer of a thymic stromal cell clone, MRL104.8a, that induces growth-maintenance and differentiation of such thymocytes. The involvement of FN-FN receptor interaction in this adhesion was demonstrated by the following lines of evidence: (i) the MRL104.8a cells expressed FN molecules on their surface and (ii) the adhesion of CD4-8- thymocytes to MRL104.8a monolayers was almost completely inhibited by simultaneous addition of anti-FN antibody and a mixture of peptides (V10 plus RGDS) capable of binding to anti-FN receptors (VLA-4 and -5). Most important, blocking the adhesion of CD4-8- thymocytes to the thymic stromal cell monolayer resulted in potent inhibition of the differentiation of these thymocytes, which was otherwise induced toward the expression of CD4 and/or CD8 molecules. These results indicate that immature CD4-8- thymocytes adhere to thymic stromal cells preferentially through FN-FN receptor interaction and that such adhesion has a critical role in inducing and/or supporting the differentiation of these thymocytes.

Animals↗

PubMed1991

Tetrahymena 14-nm filament-forming protein has citrate synthase activity.

The Tetrahymena 14-nm filament-forming protein (49K protein) is a structural protein involved in oral morphogenesis and in pronuclear behavior during conjugation. Cloning the 49K protein gene from a Tetrahymena thermophila cDNA library, we found that its primary structure exhibits a high sequence identity (51.5%) with porcine heart citrate synthase and retains functional domains. The 49K protein actually possesses citrate synthase activity, and is detected in mitochondria. These results suggest that the 49K protein has dual functions as both a respiratory enzyme and a structural protein in the cytoskeleton.

Amino Acid Sequence↗

PubMed1991

Effect of tunicamycin, an inhibitor of protein glycosylation, on testicular cord organization in fetal mouse gonadal explants in vitro.

The effect of tunicamycin (TM) on testicular cord organization in the fetal mouse was examined in vitro at light and electron microscopic levels, with special reference to the glycoprotein functions during Sertoli cell differentiation. In testicular explants treated with TM, testicular cord organization was inhibited. TM treatment affected basal lamina formation by Sertoli cells, resulting in a discontinuous basal lamina or none at all in certain areas. The disorganized Sertoli cells were amorphous in shape, exhibited poor epithelial polarity, and were irregularly arranged in the testicular parenchyma. Extracellular matrix and collagen fibers were often observed in the intercellular spaces between the disorganized Sertoli cells. Lectin histochemical observation revealed that the number of wheat germ agglutinin binding sites on the plasma membrane and basal lamina of disorganized Sertoli cells was significantly decreased by TM treatment. However, junctions were normally observed in the plasma membrane between disorganized Sertoli cells. Leydig cells showed a normal differentiation in the testicular parenchyma in the presence of TM. These observations suggest that basal lamina formation of Sertoli cells and/or the expression of their cell surface glycoconjugates may be crucial for the establishment of Sertoli cell polarity and/or the Sertoli-Sertoli cell interactions required for proper testicular cord formation. Sertoli cell organization into testicular cords and Leydig cell differentiation may be controlled by different regulatory mechanisms.

Animals↗

PubMed1991

Reconstruction of basement membrane in recombinants of epidermis and dermis of chick embryonic skin in vitro: an electron microscopic study.

The tarsometatarsal skin from 13-day-old chick embryos was treated with EDTA and/or Dispase to separate it into epidermis and dermis, and the basal lamina was removed. The isolated epidermis and dermis were then recombined and cultured on Millipore filters in a chemically defined medium (BGJb). Beginning at 3-4 days after recombination, short fragments of new basal lamina and subbasal dense plaque were formed along the epidermal basal cell outer surface immediately subjacent to hemidesmosomes. After 6-8 days of culture, fragments of the basal lamina started to fuse together and the lamina became progressively continuous. At the same time, anchoring fibrils were formed to attach to the basal lamina. The hemidesmosome formation preceded the basement membrane formation. When normal embryonic epidermis was recombined with retinol-pretreated dermis and cultured for 7 days in BGJb, short fragments of the basal lamina, the subbasal dense plaque, and anchoring fibrils were formed, but the basement membrane remained discontinuous with many interruptions in the interspace between hemidesmosomes. These results demonstrate that pretreatment of dermis with retinol causes the changes noted in the basement membrane.

Animals↗

PubMed1991

A protein in the oxygen-evolving complex in the chloroplast is associated with symptom expression on tobacco leaves infected with cucumber mosaic virus strain Y.

To elucidate the molecular basis of symptom expression in virus-infected plants, the changes in proteins between tobacco, Nicotiana tabacum cv. Ky57, leaves inoculated with cucumber mosaic virus strain Y [CMV(Y)] and strain O [CMV(O)], were compared by 2-dimensional (2-D) gel electrophoresis. The appearance of chlorotic spots in CMV(Y)-inoculated tobacco leaves accompanied an increase of 3 polypeptides and a decrease in 6 polypeptides, as compared with those in the CMV(O)-inoculated tobacco which showed no clear symptoms. The decrease in the amounts of two polypeptides of 22 and 23 kDa was particularly significant: these two polypeptides were compared with a 24 kDa polypeptide, which co-migrated with them in 2-D gel electrophoresis but did not clearly decrease at an early stage of infection, as well as major other proteins of CMV(Y)-inoculated tobacco leaves. However, the 22, 23 and 24 kDa polypeptides showed the same peptide mapping pattern. Furthermore, the 12 amino acid residues at N-termini of the three polypeptides match those of the extrinsic 23 kDa polypeptide of an oxygen-evolving complex from spinach. A comparative analysis of the 22, 23 and 24 kDa polypeptides in N. tabacum and its ancestral parents, N. sylvestris and N. tomentosiformis, revealed that the 22 kDa polypeptide derives from N. sylvestris and the 23 kDa polypeptide from N. tomentosiformis; the 24 kDa polypeptide derives from both ancestral Nicotiana species. The results indicate that the polypeptides whose amounts differentially decrease with the progress of symptom expression in N. tabacum inoculated with CMV(Y) are one component of the oxygen-evolving complex in photosystem II.

Amino Acid Sequence↗

PubMed1991

Stimulation by Bt2cAMP of epidermal mucous metaplasia in retinol-pretreated chick embryonic cultured skin, and its inhibition by herbimycin A, an inhibitor for protein-tyrosine kinase.

Epidermal mucous metaplasia of cultured 13-day-old chick embryonic tarsometatarsal skin can be induced by culture in medium containing retinol (20 microM) for only 8-24 h and then in a chemically defined medium without vitamins or serum for 6 days. In the induction of mucous metaplasia, retinol primarily affects the dermal cells and a signal(s) induced in the dermis by excess retinol alters epidermal differentiation toward secretory epithelium. In this work we found that Bt2cAMP (2 mM) stimulated mucous metaplasia severalfold when added to retinol-pretreated skin but inhibited epidermal mucous metaplasia when added together with retinol. Forskolin (100 microM), an activator of adenylate cyclase, also stimulated mucous metaplasia when added to retinol-pretreated skin. On the other hand, transduction in the epidermal cells of a signal(s) induced in dermal cells by excess retinol was inhibited by herbimycin A (500 ng/ml), an inhibitor of protein-tyrosine kinases, and TPA (0.1 microM), an activator of protein kinase C. Hence these findings indicated that cAMP stimulated signal-induced mucous metaplasia, and that transduction of the signal(s) in the epidermal cells required protein-tyrosine kinase and was inhibited by protein kinase C.

Adenylyl Cyclases↗

PubMed1991

Demonstration of beta amyloid protein-containing neurofibrillary tangles in parkinsonism-dementia complex on Guam.

This report concerns the immunohistochemical demonstration of beta amyloid protein-containing neurofibrillary tangles (beta-NFTs) from patients with parkinsonism-dementia complex on Guam (PDC). Formalin-fixed paraffin-embedded tissue sections from the temporal lobe of nine PDC cases and the spinal cord of three were examined. An affinity-purified antibody to the N-terminal peptide sequence of beta amyloid protein was used. beta-NFTs were demonstrated in the temporal lobes of the three oldest individuals. Some senile plaques, probably a manifestation of ageing, were also seen. The beta-NFTs were most frequent in the entorhinal cortex. Senile plaques were invariably present in the vicinity of the beta-NFTs. Conversely, in the regions without senile plaques, even though numerous neurofibrillary tangles were identifiable with the modified Bielschowsky method, no beta-NFTs were seen. The density of beta-NFTs appeared to be proportional to that of senile plaques, none was seen in cases lacking senile plaques. No beta-NFTs were detected in the spinal cords. Our data suggest that beta-NFTs result from secondary beta amyloid deposition on certain neurofibrillary tangles.

Amyloid beta-Peptides↗

PubMed1991

Plasma thrombosis-inducing activity in 120 patients with primary lung cancer.

One hundred and twenty patients with primary lung cancer were examined for the presence of thrombosis-inducing activity (TIA) in their plasma. TIA was identified in plasma from 16 of 38 patients with stage 3 (42%) and 31 of 65 patients with stage 4 (48%) disease. On the other hand, only 1 of 17 patients with stages 1 and 2 (6%) showed TIA in their plasma. Cell type did not seem to correlate with the presence of plasma TIA, since TIA was identified in plasma from patients with all cell types. Survival of 32 patients with inoperable non-small cell lung cancer, all stage 4, was studied. The mean survival time was 7.2 months in the TIA-positive group and 10.3 months in the TIA-negative group. This difference was statistically significant.

Adult↗

PubMed1991

Relationship between the duration of fetal breathing movements and gestational age and the development of the central nervous system at 25-32 weeks of gestation in normal pregnancy.

In 15 pregnant women with normal pregnancy at 25-32 weeks of gestation, fetal breathing movements (FBM), fetal body movements and fetal heart rate were continuously and simultaneously recorded over a total period of 180 min. The frequency of successive FBM with a duration less than 10 s (apnea period greater than 3 s) showed a significant decrease (p less than 0.01), while the frequency of successive FBM with a duration greater than or equal to 30 s showed a significant increase from 25 to 32 weeks of gestation (p less than 0.001). Strong positive correlations were also demonstrated between the frequency of FBM with a duration of greater than or equal to 30 s and the number of fetal heart rate accelerations per hour, acceleration/fetal body movement ratio, and the value of fetal heart rate long-term variability. These results suggest that the length of the duration of successive FBM represents a useful parameter for the analysis of fetal respiratory patterns, and that a prolongation of the duration of successive FBM of an individual fetus is related to the functional development of the central nervous system.

Central Nervous System↗

PubMed1991

Relationship of fetal breathing movement pattern to surfactant phospholipid levels in amniotic fluid and postnatal respiratory complications.

Fetal breathing patterns in 102 patients delivering before 37 weeks of gestation were divided into three types by the duration of successive respiratory activity using an ultrasonic scanner. Approximately 80% of the fetuses with fetal breathing absent or less than 9 s developed neonatal respiratory complications, including 11 cases of severe RDS and 8 cases of apnea of prematurity. However, no infant showing breathing activity of 30 s or more in utero experienced severe respiratory complications after birth. Furthermore, the surfactant phospholipid concentration in amniotic fluid was significantly increased with prolongation of successive fetal breathing activity. These results suggest that the duration of successive fetal breathing movements closely relates with fetal lung development including biochemical and neuromuscular systems in the humans, i.e. functional maturation of the fetal respiratory system, and also that prenatal analysis of breathing patterns may permit evaluation of pulmonary functional capacity after birth.

Amniotic Fluid↗

PubMed1991

Localization of Na(+)-dependent active type and erythrocyte/HepG2-type glucose transporters in rat kidney: immunofluorescence and immunogold study.

Glucose is actively taken up from the glomerular filtrate into the tubule cells by the Na(+)-dependent active glucose transporter (GT), and passively crosses the basolateral membrane via facilitated diffusion GT. With the use of antibodies directed against two types of GTs, we show the immunocytochemical localization of the Na(+)-dependent active GT (SGLT1) and the erythrocyte/HepG2-type facilitated diffusion GT (GLUT1). For light microscopic observation, frozen sections were stained by the rhodamine labeling method. Counterstaining with fluorescein-phalloidin and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) was employed to facilitate cell type identification. Immunogold staining was carried out on ultra-thin frozen sections for electron microscopy. The antibody to SGLT1 reacted with a 77 KD protein in immunoblotting of a kidney lysate. By immunocytochemistry, SGLT1 was localized in the microvillous plasma membrane in the apical brush borders of the cells of all three proximal tubule segments (S1, S2, and S3). The antibodies to GLUT1, a member of the facilitated diffusion GT family, were raised against human erythrocyte GT or synthetic oligopeptides derived from HepG2 GT, which reacted with a 48 KD protein in immunoblotting of the kidney lysate. GLUT1 was found at the basolateral plasma membranes of S3 proximal tubule cells, cells of the thick limb of Henle's loop, and collecting duct cells. Combined with known physiological data, our findings suggest that SGLT1 in the apical plasma membrane of the proximal tubule cells is responsible for the Na(+)-dependent active reabsorption of glucose from the glomerular filtrate. GLUT1 in the basolateral plasma membrane of S3 cells may transport reabsorbed glucose to the blood vessels. GLUT1 in the basolateral plasma membranes of cells of the thick limb of Henle's loop and of the collecting duct, on the other hand, may nourish these metabolically active cells by facilitating the diffusion of extracellular glucose provided from blood through the basolateral side of the cells.

Animals↗

PubMed1991

Cloning and sequencing of viral integration site in human fibroblasts immortalized by simian virus 40.

We have analyzed cellular DNA sequences at the viral genome integration site in a human fibroblast cell line VA13 immortalized by simian virus 40 (SV40). The computer analysis of the junctional cellular DNA sequences did not show any homology to the DNA sequences previously reported. This suggests that immortalization by SV40 was not induced by the destruction of any known oncogene or anti-oncogene at the integration site. We did not find the precise substantial sequence homology at the junctional site between the cellular DNA and SV40 DNA, indicating that the recombination mechanism involved does not require precise sequence homology and therefore, SV40 genome was probably not integrated by homologous recombination. Short direct and inverted repeats of 5 to 29 nucleotides were found in the junctional cellular and SV40 DNA. Cellular DNA abutting SV40 DNA was found by the Northern blot analysis to be expressed in diploid human fibroblasts and SV40-transformed cells. The nature of this RNA is now under study.

Base Sequence↗

PubMed1991

Step section preparation of transbronchial lung biopsy. Significance in the diagnosis of diffuse lung disease.

We retrospectively prepared step sections of nondiagnostic TBLB materials obtained from patients with diffuse or multiple lung disease and evaluated the diagnostic significance of the step section method. Among 112 patients with nondiagnostic TBLB findings, the preparation of step sections resulted in specific findings in seven cases. Step sections were especially useful for the detection of epithelioid granulomas and tumor tissue in patients with sarcoidosis and lymphangiosis carcinomatosa, respectively, but their contribution to the diagnosis of hypersensitivity pneumonitis, collagen-vascular disease, Mycoplasma pneumonia and pneumoconiosis was relatively small. In addition, step sections were useful for the detection of bronchiolitis obliterans affecting respiratory bronchioles. Overall, the examination of step sections was considered to be clinically useful in 30 cases (26.8 percent). Accordingly, the examination of step sections can be recommended before a further diagnostic procedure is chosen, if a TBLB performed in patients with diffuse or multifocal lung disease is nondiagnostic.

Biopsy↗

PubMed1991

The amino-acid sequences of the Bacillus stearothermophilus ribosomal proteins S17 and S21 and their comparison to homologous proteins of other ribosomes.

Ribosomal proteins S17 and S21 from the moderate thermophile Bacillus stearothermophilus were purified by one-step high-performance liquid chromatography from the 30S-subunit protein mixture employing a semi-preparative reversed-phase C4 column. The complete amino-acid sequences of these proteins were determined by a combination of N-terminal sequencing in picomole quantities of the protein and of appropriate peptide fragments. Proteins S17 and S21 consist of 86 and 55 amino-acid residues, corresponding to molecular masses of 10074 and 6593 Da, respectively. They are homologous to proteins S17 and S21 from the Escherichia coli ribosome, showing 50 and 55% identities in the corresponding regions, respectively. The C-terminal region of protein S21 from B. stearothermophilus has a deletion of 15 residues as compared to the E. coli S21 protein. The evolutionary relationships of the Bacillus proteins to various other members of the S17 and S21 ribosomal protein families are discussed.

Amino Acid Sequence↗

PubMed1991

[Neurosurgical management of dialyzed patients].

Cerebrovascular disease is a lethal complication for patients with renal failure because of the hemostatic disturbance and equivocality about management of the central nervous system. Appropriate management of the renal failure in neurosurgical patients was considered on the basis of experience. Hemodialysis (HD), which is the most common dialysis method, has a serious disadvantage: the elevation of intracranial pressure during dialysis due to the "disequilibrium syndrome." It is important to stabilize the serum osmolarity during dialysis in order to prevent the disequilibrium syndrome. From this point of view, continuous ambulatory peritoneal dialysis (CAPD) has great advantages; serum osmolarity is not rapidly changed and no anticoagulants are required during dialysis. CAPD is recommended as the first method of choice in neurosurgical management of renal failure patients. However, if a patient has to be maintained with HD because of a history of laparotomy or peritonitis, it is essential to keep the serum osmolarity as stable as possible using the extracorporeal ultrafiltration method, hypernatremic HD, bicarbonate HD, and intravenous administration of glycerol.

Adult↗

PubMed1991

An improved fixation method for transmission electron microscopy for the histological study of the lens.

An improved fixation technique for transmission electron microscopic observation that enables good fixation of all areas of the rat lens was devised. Immersion fixation with 0.1 M phosphate-buffered 1.25% glutaraldehyde at 4 degrees C for 12 hours followed by postosmication produced good results for all areas of the lens--anterior, equatorial, and posterior zones. The technique was particularly suitable for maintenance of the shape of the lens since practically no irregularity, vacuolar degeneration, or expansion of the intercellular spaces was noted. This technique, which requires only a relatively short time, was also useful for the detection of early ultrastructural changes associated with cataract in spontaneously diabetic WBN/Kob rats. We anticipate that our procedure will be widely applied.

Animals↗