Biomedical subjects

N Hashimoto

Publications and source records attributed to N Hashimoto.

At least 415 records · Page 23Linked to original sources

PubMed1993

Differential contribution of M(r) 120 kDa rasGTPase-activating protein and neurofibromatosis type 1 gene product during the transition from growth phase to arrested state in human fibroblasts accompanied by a unique rasGTPase-activating activity.

Using octyl glucoside-solubilized cell extracts from human fibroblasts during growth phase to G0/G1 arrest state, we found that while the number of M(r) 120 kDa rasGTPase-activating protein (p120GAP) molecules per cell decreases to half its original levels, the amount of neurofibromatosis type 1 gene product (NF1, a neurofibromin) remains constant during the transition. The contribution of p120GAP to the total rasGTPase-activating (rasGA) activity in growing cells was found to be larger than that observed in arrested cells (84% vs 53%). On the other hand, NF1 contributes less than 15% of the total rasGA activity in either extract. These results indicate that the qualitative changes occur in the contributors to rasGA activity during transition. They also suggest that a unique rasGA activity exists in the arrested cells, which was obtained separately from both p120GAP and NF1 by heparin-Sepharose column chromatography.

Animals↗

PubMed1993

Common sequence elements are important for transcription and replication of the extrachromosomal rRNA-encoding genes of Tetrahymena.

Three types of conserved sequence elements have previously been identified in the generally non-conserved, 5' non-transcribed spacer region of the rRNA-encoding gene (rDNA) of Tetrahymena species. In vivo and in vitro experiments have identified the start point (tsp) for rDNA transcription. Comparative sequence analysis suggested that the type-I sequence element was likely to function as the promoter for rDNA transcription. We have used in vitro transcription assays to demonstrate that sequences in the 3' half of the type-I repeat located proximal to the tsp are essential for the accurate initiation of rRNA synthesis. These sequences define the 5' boundary of the core or minimal promoter element and include the sequences which function in rDNA replication and maintenance in the homologous regions in two upstream type-I sequence elements. These sequences may bind factors important for both the transcription and replication of rDNA.

Animals↗

PubMed1993

[Usefulness of enhanced MR angiography by nitroglycerin].

We tried to use nitroglycerin to enhance MR angiography (MRA). Nitroglycerin has an effect to dilate vessels. We made a direct comparison between pre and post enhanced MRA (3 D-TOF technique). MRA after administration of nitroglycerin could depict more details of cortical branches without enhancement of background. So we recommend this method to be used in any examination of MR angiography.

Adult↗

PubMed1993

Improved sensitivity in the antibody-forming cell assay for Japanese encephalitis virus in mice by optimal fixation of cells and avidin-biotin complex (ABC) immunocytochemistry.

A sensitive modified method for the detection in vitro of antibody-forming cells (AFCs) in Japanese encephalitis (JE) virus infected mice is described. The procedure involves the selection of the most appropriate incubation period, after the infection of monolayer cells, for expression of viral antigens to be recognized by AFCs. Adequate preservation of the expressed antigens was achieved after fixation with paraformaldehyde-lysine-periodate buffer (PLP). The development of spots which corresponded to AFCs was successfully obtained with the use of the avidin-biotin complex (ABC) amplification reaction. The number of the spots developing after completion of the assay was greater after PLP fixation compared with other fixation solutions such as methanol, formalin and acetone-water. Optimum antigen expression on the infected cells for recognition by lymphoid cells was reached after 24 h of infection. However, it was possible to detect viral antigens on the infected cells 8 h after infection. The ABC reaction was shown to be the best procedure for developing spots compared with other immunocytochemical methods. A linear increment was observed in the number of spots that developed at different spleen cell densities. This assay is simple to perform and could detect virus-specific Ig producing cells from the spleens of mice infected with JE virus. It also makes it possible to enumerate accurately the kinetics of the AFC response in different lymphoid organs during infection or after vaccination.

Animals↗

PubMed1993

A novel murine zinc finger gene mapped within the tw18 deletion region expresses in germ cells and embryonic nervous system.

A novel zinc finger gene, designated NT fin12, belonging to the C2H2-Krüppel-type gene family was isolated from a newborn mouse testis cDNA library by using zinc finger consensus motif probes. Northern blot analyses showed that NT fin12 mRNA was expressed during the meiotic prophase of spermatogenesis and in embryogenesis. Transcripts were localized by in situ hybridization in spermatogonia and in early spermatocytes, and in testis cords in the genital ridge as well as in oocytes and follicle cells in the ovary. In midgestational embryos at 8.5-13.5 days postcoitum, transcripts were present in the neuroectoderm, and they were progressively restricted to peripheral ganglia derived from neural crest cells and neural placodes and to the motor nerve cells in the central nervous system. Taken together these results indicate that NT fin12 functions during germ cell development and also plays a role in the specification of a subpopulation of neuroectodermal cells. Genetic linkage analyses revealed that the NT fin12 locus mapped to the deletion region of the tw18 haplotype on mouse chromosome 17.

Amino Acid Sequence↗

PubMed1993

A comparison of the metabolic changes after the distal splenocaval and portacaval shunts.

The distal splenorenal shunt (DSRS) is the recommended procedure for selective variceal decompression, but its use may be limited in patients who have undergone left nephrectomy, those with an anatomically aberrant relationship between the splenic and left renal veins, and those whose preoperative angiographic findings suggest a risk of developing postoperative renal vein hypertension. For these clinical situations, the selective distal splenocaval shunt (DSCS) is a useful alternative to the DSRS. However, the metabolic consequences of the DSCS have not yet been studied in detail and therefore, using a canine model, the metabolic changes following the DSCS and the portacaval shunt (Eck) were compared. The metabolic changes observed following the Eck were hyperammonia and amino acid imbalance, while those following the DSCS were similar to those of the control dogs. In terms of the hepatic adenosine triphosphate level, which reflects hepatic mitochondrial function, the DSCS dogs were also similar to the control dogs. These data suggest that there was no metabolic disadvantage of the DSCS compared to the control.

Adenosine Triphosphate↗

PubMed1993

Type 2 (non-insulin-dependent) diabetes mellitus associated with a mutation of the glucokinase gene in a Japanese family.

Mutations were screened for in the glucokinase gene of 25 Japanese patients with Type 2 (non-insulin-dependent) diabetes mellitus. Each exon was scanned by electrophoresis of enzymatically amplified DNA segments under non-denaturing conditions and variants were sequenced. A variant pattern was detected in exon 5 of one patient. Direct sequencing of this exon revealed a single nucleotide substitution in codon 188 (GCT-->ACT) of one of two alleles resulting in the mutation of Ala188-->Thr, an invariant residue in the sequence of all mammalian glucokinases and hexokinases. This mutation was not found in 40 normal control subjects. The proband had been diagnosed with Type 2 diabetes at the age of 62 years. Four other members of her family have the same mutation and all have Type 2 diabetes or impaired glucose tolerance. The youngest age at diagnosis of Type 2 diabetes in these other members was 13 years, suggesting that her pedigree was maturity-onset diabetes of the young (MODY). All subjects with the Thr188 mutation show a decreased insulin secretory response during oral glucose tolerance testing. Mutations in the glucokinase gene associated with Type 2 diabetes have been previously identified in Caucasian (French and British) subjects. This study indicates that mutations in this gene are also implicated in the development of Type 2 diabetes in Asians. Further studies are required to determine the frequency of mutations in glucokinase among Japanese patients with Type 2 diabetes.

Adolescent↗

PubMed1993

Degenerative changes in the internal elastic lamina relating to the development of saccular cerebral aneurysms in rats.

In order to investigate the developmental mechanism of saccular cerebral aneurysms, changes in the internal elastic lamina at the junction of the anterior cerebral artery and the olfactory artery were electronmicroscopically studied in 6 control and 6 experimental rats undergoing ligation of the left carotid artery and branches of both renal arteries. In the control group, spontaneous destructive changes occurred on the luminal side of the internal elastic lamina and progressed from the luminal towards the abluminal side as the elastic lamina advanced to the apex. Close to the apex, these changes invaded and disrupted the whole elastic lamina. The elastic lamina was replaced by sparsely lined up lumps of elastic tissue in the walls of early aneurysmal alterations, and was atrophied and disappeared totally in the walls of aneurysmal alterations that had reached an advanced stage. These spontaneous changes were in agreement with reports in the literature and our own previous investigations. From the findings in the experimental rats it becomes likely that the aneurysmal changes in the elastic lamina are exaggerated forms of the normal catabolic metabolism. Therefore its synthesis on the abluminal side no longer balances with the catabolism on the luminal side. It is strongly suggested that aneurysmal alterations progress from the luminal towards the abluminal side of arterial walls and that the lytic process of elastase might play a role in the degenerative changes in aneurysmal development.

Animals↗

PubMed1993

In vivo flow visualization of induced saccular cerebral aneurysms in rats.

A surgical procedure to expose the arterial bifurcation at the base of the rat brain was developed without sacrificing the animal. Using this technique, visualization of flow in and around the induced cerebral aneurysm was achieved by detecting and following fluorescent particles in the blood stream. Cerebral aneurysms were produced by ligating one common carotid artery, inducing experimental hypertension and feeding them with beta-aminopropionitrile. Flow studies of the arterial bifurcation with an early aneurysmal formation showed that there were spiral flows proximal and distal to the bifurcation. This was the first direct visualization of the actual flow in and around cerebral aneurysms in a vital state. This technology can add further information on the development, growth and rupture of cerebral aneurysms.

Animals↗

PubMed1993

Neutralization of Chlamydia psittaci with monoclonal antibodies.

Neutralization of Chlamydia (C.) psittaci avian strain P-1041 was examined in vitro using monoclonal antibodies (MAbs). Of the 10 MAbs used, 6 were found to exhibit neutralizing capability. These include 3 against major outer membrane protein (MOMP), 1 against lipopolysaccharide (LPS) and 2 against other protein molecules [90 kilodalton (kDa) and 90/50 kDa]. Most neutralizing MAbs were dependent on complement for efficient neutralization, while a strain-specific MAb (2B5) against the 90 kDa protein displayed a different requirement for complement and neutralized the infectivity of the P-1041 at high concentrations without complement. By competitive inhibition enzyme-linked immunosorbent assay (competitive inhibition ELISA), all 3 neutralizing anti-MOMP MAbs were demonstrated to recognize different epitopes found in very close proximity to each other on the outer membrane.

Antibodies, Bacterial↗

PubMed1993

Japanese patients with chronic fatigue syndrome are negative for known retrovirus infections.

Although chronic fatigue syndrome (CFS) is known to be the syndrome that begins with an acute flu-like illness that may be due to the exposure to an infectious agent, there has been no convincing evidence on the causative agents. Recently, human T-lymphotropic virus type II (HTLV-II)-like virus has been reported to be associated with the CFS by using HTLV Western blot analysis and polymerase chain reaction. However, some investigators could not detect HTLV-II by indirect immunofluorescence analysis. Lately, CFS patients have been reported in Japan. We detected all 30 tested patients with CFS were seronegative for HTLV-II, HTLV-I and HIV by specific peptide ELISA and Western blot. Further, PCR analysis was negative for HTLV-II and retrovirus was not detected by coculture method with patients' PBMC. Thus, known human retrovirus infections do not cause a CFS in Japan.

Blotting, Western↗

PubMed1993

Moebius syndrome associated with pituitary dwarfism and hypoplastic optic disc.

A 17 year old male patient with Moebius syndrome with pituitary dwarfism and unilateral hypoplastic optic disc is presented. Although there have been several reports of an association of Moebius syndrome and pituitary dysfunction, growth hormone deficiency has not been reported previously. These associations may give some insight into the pathogenesis of Moebius syndrome.

Adolescent↗

PubMed1993

Antimicrobial effects of a new carboxyquinolone drug, Q-35, on five serogroups of Leptospira interrogans.

New carboxyquinolone drugs, including the recently developed Q-35, were evaluated for their in vitro potency against five serogroups of Leptospira interrogans. Q-35, ofloxacin, ciprofloxacin, and tosufloxacin showed MICs (0.05 to 0.20 microgram/ml) comparable to those of tetracycline. However, MBCs of these drugs varied between 10- and 100-fold above the MIC for most strains tested. Q-35 was shown to be active against L. interrogans in vitro as judged by the MICs obtained.

Anti-Infective Agents↗

PubMed1993

A glycine-1008 to valine mutation in the insulin receptor in a woman with type A insulin resistance.

We examined the insulin receptor gene in a Japanese woman with type A insulin resistance. Acanthosis nigricans and polycystic ovary were present. A 75-g oral glucose tolerance test showed a diabetic pattern, and fasting insulinemia was 780 pmol/L. Insulin binding was normal, but autophosphorylation and tyrosine kinase activity were reduced in partially purified insulin receptors from Epstein-Barr virus-transformed lymphocytes. The nucleotide sequence for all 22 exons of the insulin receptor gene was determined by direct sequencing of genomic DNA amplified with the polymerase chain reaction. Substitution of valine for glycine at codon 1008 in the tyrosine kinase domain was identified in one allele. This was the same mutation found in another patient, but there was no relationship between the two families. The father had the same mutation in one allele and impaired glucose tolerance with mild hyperinsulinemia, but the mother and two brothers had normal glucose tolerance. We conclude that a single mutant allele in the tyrosine kinase domain caused the insulin resistance.

Adult↗

PubMed1993

Statistical test of the shortening of latent periods in experimental radiation carcinogenesis under competing risks.

The mean life span of animals killed by a specific type of tumor is influenced by competing risks. The parametric inference method of Noda et al. was applied to a set of mouse experimental data in order to correct for the appearance of observed life span distribution under competing risks and to test statistical significances of difference between corrected mean life spans. Data were from an experiment on tumors induced in mice by X-rays: whole body irradiated with 600 R; head with 800 R; trunk or lower body with 800 R. The statistically significant changes were found: Whole body irradiation shortened the mean latent period of malignant lymphomas whereas the head irradiation shortened the latent periods of none of tumors. Trunk irradiation shortened mean latent periods of lung tumors. Lower body irradiation showed a tendency to shorten the mean latent periods of mammary tumors. Results that differed from the above were obtained when there was no correction for competing risks.

Animals↗

PubMed1993

Seroepidemiological survey of chlamydial infections in light horses in Japan.

To investigate the overall prevalence of chlamydial infections in light (i.e. non-draught) horses in Japan, 599 sera obtained from 12 localities in 1991 were tested for complement fixation antibodies. The mean antibody positive rates of the all sera were 15.2% (91/599) and the regional positive rates were higher in Honshu (19.1%, 48/251) and Kyushu (20.0%, 20/100) than in Hokkaido (9.3%, 23/248). In Honshu, the highest rate (56.0%, 28/50) was observed in Utsunomiya. Analysis of the positive rate in different age groups showed that the 2-5 years age-group had the highest prevalence of chlamydial infections. This indicates that chlamydial infection is prevalent in light horses in Japan.

Animals↗

PubMed1993

Evaluation of diagnostic assays for neonatal and infantile chlamydial infections.

Several diagnostic assays for neonatal and infantile chlamydial infections, isolation with tissue culture, antigen detection by enzyme immunoassay (EIA) (IDEIA Chlamydia test), a nonisotopic DNA probe (Gen-Probe PACE 2 assay), serum IgM antibody detection by EIA (SeroELISA Chlamydia TRUE IgM), and polymerase chain reaction (PCR), were evaluated. Of 210 clinical specimens (170 nasopharyngeal and 40 conjunctival swabs) from 53 neonates and 102 infants with respiratory insufficiency and respiratory tract infections which were suspected to be associated chlamydial infection, chlamydial antigens were detected in 30 by IDEIA Chlamydia. Of these 30 samples, C. trachomatis was isolated from 27 specimens. Samples from 15 neonates and 6 infants were culture-positive and IDEIA Chlamydia-positive. Of 30 samples, 27 were tested with PCR and 8 with DNA probe. Twenty-three of 27 specimens were positive with PCR, while only one specimen was positive with DNA probe. EIA can be used for the diagnosis and screening of neonatal and infantile chlamydial infections.

Antibodies, Bacterial↗

PubMed1993

An immunohistochemical study on the gastrointestinal endocrine cells of three honeyeaters: singing honeyeater (Meliphaga virescens), spiny-cheeked honeyeater (Acanthogenys rufogularis) and brown honeyeater (Lichmera indistincta).

The gastrointestinal endocrine cells of the singing honeyeater (Meliphaga virescens), spiny-cheeked honeyeater (Acanthogenys rufogularis) and brown honeyeater (Lichmera indistincta) were studied immunohistochemically with special reference to their degree of dependency upon nectar. The nine types of immunoreactive endocrine cells were detected in their gastrointestinal mucosa. Coexistence of motilin and serotonin in the same cells was confirmed in the pyloric region. In the duodenum and jejunum, a few peptide tyrosin tyrosin (PYY)-immunoreactive cells were detected. The clear difference in the distribution and frequency of the gastrointestinal endocrine cells among three types of honeyeaters, which differ in the degree of dependency upon the nectar, could not be confirmed. However, some differences were found that serotonin-, somatostatin- and gastrin-releasing peptide (GRP)-immunoreactive cells in the gizzard, gastrin-immunoreactive cells in the duodenum and jejunum, enteroglucagon-immunoreactive cells in the caeca and in the colon, and pancreatic glucagon-immunoreactive cells in the ileum of brown honeyeaters were more numerous (p < 0.05) than other two species.

Animals↗