Biomedical subjects

Y Honda

Publications and source records attributed to Y Honda.

At least 163 records · Page 9Linked to original sources

PubMed1999

Expression and activity of 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4-isomerase in different regions of the avian brain.

Recently, we have demonstrated, using biochemical and immunochemical methods, that the quail brain possesses the cholesterol side-chain cleavage enzyme (cytochrome P450scc) and produces pregnenolone and its sulfate ester. To clarify progesterone biosynthesis in the avian brain, therefore, we examined the expression of messenger RNA (mRNA) encoding for the enzyme 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4-isomerase (3beta-HSD) and its enzymatic activity using the quail. RT-PCR analysis together with Southern hybridization indicated the expression of 3beta-HSD mRNA in the brain of sexually mature birds but with no clear-cut sex difference. Employing biochemical techniques combined with HPLC analysis, the conversion of pregnenolone to progesterone was found in brain slices of mature males. Progesterone biosynthesis was increased in a time dependent manner and completely abolished by trilostane, a specific inhibitor of 3beta-HSD. The enzymatic activity of 3beta-HSD was greatest in the cerebrum and lowest in the mesencephalon. A specific RIA indicated that progesterone concentrations in the different brain regions closely followed the level of 3beta-HSD activity. High levels of progesterone concentration were observed in the diencephalon and cerebrum with lowest values in the mesencephalon. Progesterone levels in the brain regions were significantly higher than those in the plasma. These results suggest that the avian brain possesses not only cytochrome P450scc but also 3beta-HSD and produces progesterone. It is also indicated that progesterone biosynthesis in the avian brain may be region-dependent.

3-Hydroxysteroid Dehydrogenases↗

PubMed1999

Thermal unfolding of chitosanase from Streptomyces sp. N174: role of tryptophan residues in the protein structure stabilization.

Tryptophan residues in chitosanase from Streptomyces sp. N174 (Trp28, Trp101, and Trp227) were mutated to phenylalanine, and thermal unfolding experiments of the proteins were done in order to investigate the role of tryptophan residues in thermal stability. Four types of mutants (W28F, W101F, W227F and W28F/W101F) were produced in sufficient quantity in our expression system using Streptomyces lividans TK24. Each unfolding curve obtained by CD at 222 nm did not exhibit a two-state transition profile, but exhibited a biphasic profile: a first cooperative phase and a second phase that is less cooperative. The single tryptophan mutation decreased the midpoint temperature (Tm) of the first transition phase by about 7 degrees C, and the double mutation by about 11 degrees C. The second transition phase in each mutant chitosanase was more distinct and extended than that in the wild-type. On the other hand, each unfolding curve obtained by tryptophan fluorescence exhibited a typical two-state profile and agreed with the first phase of transition curves obtained by CD. Differential scanning calorimetry profiles of the proteins were consistent with the data obtained by CD. These data suggested that the mutation of individual tryptophan residues would partly collapse the side chain interactions, consequently decreasing Tm and enhancing the formation of a molten globule-like intermediate in the thermal unfolding process. The tryptophan side chains are most likely to play important roles in cooperative stabilization of the protein.

Calorimetry, Differential Scanning↗

PubMed1999

Apoptotic DNA fragmentation and upregulation of Bax induced by transient ischemia of the rat retina.

This study was performed to examine the involvement of apoptosis and the expression of bcl-2 family genes in ischemia-induced retinal injury. Retinal ischemia was induced in adult rats by raising the intraocular pressure to 130 mmHg for 45 min. Selective damage to the inner retina was observed 7 days after ischemia. No terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) positive cells were observed in the normal retina, but there was a significant number of TUNEL positive cells 6-48 h after transient ischemia followed by a decrease at 96 and 168 h. The number of TUNEL positive cells reached a maximum at 24 h after ischemia. DNA laddering was observed on agarose gel electrophoresis with the retinas 24 and 48 h after ischemia but not in the normal retina. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) revealed that bax gene expression did not change immediately after cessation of ischemia, but gradually increased as early as 6 h, reached a peak at 24 h, then decreased to near baseline levels at 168 h. On the other hand, bcl-2 gene expression showed no obvious changes at any time after transient ischemia. Moreover, intense Bax protein immunoreactivity was detected in the retinal sections at 24 h after ischemia although little immunoreactivity was present in the normal sections. These results suggest that apoptosis associated with the expression of Bax is involved in retinal cell loss after ischemic insult.

Animals↗

PubMed1999

Impact of residual plaque burden on clinical outcomes of coronary interventions.

In this study, we summarize the role of residual plaque burden, as determined by intravascular ultrasound, on the development of restenosis following percutaneus coronary interventions. Several clinical trials have shown that the amount of residual plaque is a consistent and independent predictor of subsequent restenosis. The impact of residual plaque burden on late lumen loss is particularly augmented by negative vessel remodeling that is commonly seen after balloon angioplasty and atherectomy. However, early evidence suggests that the importance of plaque burden also applies in the context of stenting. The cotreatment of debulking may further improve the long-term outcome of stenting by maximizing an acute lumen gain with less vessel stretching, preventing stent edge problems and possibly reducing the cell source involved in the intimal hyperplastic process. Evaluation of residual plaque burden with on-line intravascular ultrasound could lead to definitive therapies via risk stratification of the treated segments.

Angioplasty, Balloon, Coronary↗

PubMed1999

Temporal profile of visual evoked responses to pattern-reversal stimulation analyzed with a whole-head magnetometer.

Magnetoencephalography (MEG) has become accepted as a useful method for non-invasively studying brain functions, including visual perception. The present study used MEG to elucidate information processing following pattern-reversal stimulation by analyzing the origins and properties of visual evoked magnetic fields (VEFs). The VEFs of ten healthy adults were recorded in a magnetically shielded room using a 122-channel whole-head magnetometer. The visual stimulation of checkerboard-pattern reversal at 1.7 Hz was presented to the subject's right hemifield. Visual evoked potentials (VEPs) were recorded simultaneously, and 150 responses were each averaged for VEFs and VEPs. For the contrast profile study, pattern-reversal stimuli at five different contrast levels from 96% to 8% were used. In all subjects, the VEFs showed three components with latencies of approximately 95, 120, and 160 ms. The equivalent current dipoles for the first and the third components were located and were oriented close to each other in the left occipital lobe, but these dipoles were separated from that of the second component, which showed an opposite direction. Stimuli at a moderate contrast level markedly reduced the first component, but not the third. These findings indicate that the first and the third components of VEFs appear to originate from anatomically closely situated, almost identical, sources, but that their physiological properties differ.

Adult↗

PubMed1999

Neuroendoscopic surgery for specific forms of hydrocephalus.

Neuroendoscopic surgery was used to treat patients with various forms of hydrocephalus with specific pathophysiology, including long-standing overt ventriculomegaly in adulthood (LOVA), isolated unilateral hydrocephalus (IUH), isolated IV ventricle (IFV), disproportionately large IV ventricle (DLFV), isolated rhombencephalic ventricle (IRV), isolated quarto-ventriculomegaly (IQV), dorsal sac in holoprosencephaly (DS), and loculated ventricle (LV). A total of 26 operative procedures were performed, with neuroendoscopic surgery in 22 patients, 12 with unique forms of noncommunicating hydrocephalus and 10 with various types of postshunt isolated compartment. These procedures included III ventriculostomy, aqueductal plasty by both rostral and caudal approaches, foraminal plasty in the foramen of Monro/foramen of Magendie, septostomy, IV ventriculostomy, fenestration of septation in the loculated ventricle, fenestration of arachnoid cyst or cystic tumor obstructing a ventricle with or without tumor removal, and dorsal sac ventriculostomy. The characteristics of the cerebrospinal fluid (CSF) dynamics in the individual specific pathophysiologies were delineated by cardiac-gated cine-mode magnetic resonance imaging (MRI) before and after the endoscopic procedure. The consequent success rate (success = restoration of communication of the CSF pathway in the individual patients) was 19/22 (86.4%). The progression of ventricular dilatation was stopped in 17 of 19 patients (89.5%) in whom the endoscopic procedure was successful (radiologically arrested hydrocephalus). Improvement in the clinical symptoms and signs (clinically arrested hydrocephalus) was obtained in 15 of the patients (68.2% of all patients: 5 with LOVA, 3 with IQV, 5 with IUH and 2 with LV). Seven patients (2 LOVA, 2 IFV, 1 DS, 1 DFLV and 1 IRV) underwent a shunt procedure after the neuroendoscopic procedure(s). The postoperative changes of ventriculomegaly were complicated, reflecting the differences in the brain parenchymal compliance and postoperatively corrected CSF flow dynamics in the major CSF pathway.

Adult↗

PubMed1999

Sequence analysis of RNA-2 of different isolates of broad bean wilt virus confirms the existence of two distinct species.

The complete nucleotide sequence of RNA-2 from a Japanese isolate IP of broad bean wilt virus (BBWV) was determined. The sequence encodes a single large polyprotein, which contains a putative movement protein and two coat proteins (CPs). The 3'-terminal sequences of RNA-2 were also determined for three other Japanese isolates and two ATCC isolates (PV132 and PV176) of BBWV. The CPs of the four Japanese isolates share 86.8-98.0% amino acid sequences homology with one another and 88.3-96.5% with those reported for the isolate PV131 (BBWV-2). However, they have only 57.9-66.2% homology with those of PV132 and PV176 (BBWV-1).

3' Untranslated Regions↗

PubMed1999

Expression of thrombospondin-1 in ischemia-induced retinal neovascularization.

Thrombospondin-1 is an extracellular matrix protein that inhibits endothelial cell proliferation, migration, and angiogenesis. This study was performed to investigate the role of thrombospondin-1 in ischemic retinal neovascularization. In a murine model of retinal neovascularization, thrombospondin-1 mRNA was increased from postnatal day 13 (P13), with a threefold peak response observed on P15, corresponding to the time of development of retinal neovascularization. Prominent expression of thrombospondin-1 was observed in neovascular cells, specifically, cells adjacent to the area of nonperfusion. It has been suggested that vascular endothelial growth factor (VEGF) plays a major role in ischemia-induced retinal neovascularization of this model, so we studied the effects of VEGF on thrombospondin-1 expression. In bovine retinal microcapillary endothelial cells, VEGF induced a biphasic response of thrombospondin-1 expression; VEGF decreased thrombospondin-1 mRNA 0.41-fold after 4 hours, whereas it increased, with a threefold peak response, after 24 hours. VEGF-induced endothelial cell proliferation was completely inhibited by exogenous thrombospondin-1 and increased by 37.5% with anti-thrombospondin-1 antibody. The present findings suggest that, in the ischemic retina, retinal neovascular cells increase thrombospondin-1 expression, and VEGF may stimulate endogenous thrombospondin-1 induction, which inhibits endothelial cell growth. VEGF-mediated thrombospondin-1 induction in ischemia-induced angiogenesis may be a negative feedback mechanism.

Animals↗

PubMed1999

Unsuspected extra-adrenal pheochromocytoma simulating ovarian tumor.

We report on a case of an extraadrenal pheochromocytoma simulating an ovarian tumor. Before intervention, the patient exhibited no symptoms suggestive of pheochromocytoma. Nevertheless, during surgery she experienced marked blood pressure fluctuations, and an unsuspected extraadrenal pheochromocytoma was diagnosed. Thus, although rare, when preparing to remove a pelvic mass, the gynecologist should consider the possibility of an extraadrenal pheochromocytoma.

Blood Pressure↗

PubMed1999

Significant association of a single nucleotide polymorphism in the tumor necrosis factor-alpha (TNF-alpha) gene promoter with human narcolepsy.

Narcolepsy is a sleep disorder in which multiple factors, including environmental and genetic factors, are involved. A genetic factor strongly associated with the disorder has been found in the human leukocyte antigen (HLA) class II region: the haplotype, DRB1*1501-DQB1*0602, predisposes to narcolepsy. No susceptibility genes other than the HLA-haplotype have been found. In this paper, we performed an association study of the tumor necrosis factor-alpha (TNF-alpha) gene located in the HLA class III region with human narcolepsy, in which we examined the known single-nucleotide polymorphisms (SNPs) in the promoter region in 49 narcoleptic patients, who were all positive for DRBI*1501, and 111 healthy control individuals. The results indicated that the frequency of the genotype at position -857 (-857SNP) was significantly different between the patients and controls, and the allele frequencies of 857SNP revealed that the frequency of -857T was significantly increased in the patients as compared with that in the controls (P=0.0068). In addition, haplotypes presumed from HLA-DRB1, -857SNP and HLA-B loci suggested that -857T was mainly associated with DRB1 alleles other than DRB1*1501: the significant increase in frequency of -857T in the patients was not caused by allelic association with DRB1*1501. Therefore, it is conceivable that the TNF-alpha with 857T was associated with narcolepsy independently of the strong association of DRB1*1501 with the disorder. Altogether, the data presented here lead us to propose that TNF-alpha could be a new susceptibility gene in human narcolepsy.

Alleles↗

PubMed1999

Tranilast inhibits protein kinase C-dependent signalling pathway linked to angiogenic activities and gene expression of retinal microcapillary endothelial cells.

1. Tranilast, first developed as an anti-allergic drug, has been reported to inhibit vascular endothelial growth factor (VEGF)-induced angiogenesis and vasopermeability. To further clarify the inhibitory mechanism, we investigated the effects of tranilast on VEGF binding and subsequent intracellular signalling pathway linked to angiogenic activities and gene expression of bovine retinal microcapillary endothelial cells. 2. Tranilast significantly (P<0.01) inhibited VEGF, basic fibroblast growth factor (bFGF), and hypoxia conditioned media-induced BREC proliferation in a dose dependent manner with IC50's of 22, 82 and 10 microM, respectively. 3. VEGF-induced migration was also inhibited by tranilast in a dose dependent manner, with IC50 of 18 microM, and complete inhibition was observed at 300 microM (P<0.01). Tranilast suppressed VEGF-induced tube formation in a dose dependent manner with maximum (46%) inhibition observed at 300 microM (P<0.05). 4. Tranilast inhibited phorbol myristate acetate (PMA)-dependent stimulation of [3H]-thymidine incorporation and VEGF- and PMA-induced gene expression of integrin alpha v and c-fos in BREC. 5. Tranilast suppressed VEGF- and PMA-stimulated PKC activity in BREC. 6. Tranilast did not affect VEGF binding or VEGF-induced phosphorylation of tyrosine residues of VEGF receptor- and phospholipase Cgamma and their associated proteins. 7. These data suggest that tranilast might prove an effective inhibitor to prevent retinal neovascularization in ischaemic retinal diseases, and that its inhibitory effect might be through suppression of PKC-dependent signal transduction in BREC.

Animals↗

PubMed1999

Colony-stimulating factors in rapid eye movement sleep and non-rapid eye movement sleep regulation.

Although several cytokines are known to be somnogenic, no study has been conducted to examine whether colony-stimulating factors (CSF) affect sleep. Therefore, we studied the effects of granulocyte-macrophage CSF (GM-CSF) and macrophage CSF (M-CSF) on sleep in rats and their possible mechanism of action. At the dose of 10 pmol, GM-CSF or M-CSF significantly increased both non-rapid eye movement and rapid eye movement (REM) sleep or REM sleep only when infused intracerebroventricularly during the dark period. When injected locally in the hypothalamus, GM-CSF and M-CSF increased nitric oxide (NO) production. Thus, NOergic neural signals in the hypothalamus may take part in the somnogenic action of CSF.

Animals↗

PubMed1999

Acetylcholine and glutamate release during sleep-wakefulness in the pedunculopontine tegmental nucleus and norepinephrine changes regulated by nitric oxide.

Cholinergic neurons in the pons appear to play a major role in generating rapid eye movement (REM) sleep. In the present study, acetylcholine and glutamate release in the pedunculopontine tegmental nucleus (PPT) during the sleep-waking cycle were investigated by in vivo microdialysis. Acetylcholine release during slow wave sleep (SWS) was significantly lower (P<0.05) than during REM sleep and wakefulness. On the other hand, glutamate release during wakefulness was higher (P<0.05) than during REM sleep and SWS. Furthermore, the application of N-methyl-D-aspartate (1 mM) induced a significant increase of nitric oxides (NOx) for 20 min (P<0.05) and a decrease of norepinephrine for the first 15 min (P=0.01), indicating NOx regulation on norepinephrine release in PPT.

Acetylcholine↗

PubMed1999

A search for a mutation in the tumour necrosis factor-alpha gene in narcolepsy.

The discovery of almost 100% association of narcolepsy with human leukocyte antigens (HLA) DR2 antigen prompted molecular biological research of this disorder. In the HLA class II gene cluster, the gene for tumour necrosis factor-alpha (TNF-alpha), which plays a role in the regulation of normal human sleep, is located. The present study searched for a mutation in the TNF-alpha gene by single-strand conformation polymorphism analysis (SSCP) in patients with narcolepsy. No mutation was detected in exons and introns of the TNF-alpha gene by SSCP and sequencing.

DNA Mutational Analysis↗

PubMed1999

Possible association of human leucocyte antigen DR1 with delayed sleep phase syndrome.

The study investigated the human leucocyte antigen (HLA), types A, B and DR, of 42 patients with delayed sleep phase syndrome (DSPS) and compared the frequencies of the antigens with those in 117 healthy controls. The comparison revealed that the gene frequencies and positivities of HLA-A, -B and -DR, except for DR1, had no significant differences between the patients and controls. The frequency of HLA-DR1 was increased in the DSPS patients as compared with that in the healthy controls (P = 0.0069 in positivity). Although the corrected P-value (0.069) for multiple comparisons almost reached the significance level, the results indicated a possible association of the HLA-DR1 antigen with DSPS. This study suggests that there are genetic predispositions to DSPS.

Adolescent↗

PubMed1999

Mutant herpes simplex virus-mediated suppression of retinoblastoma.

PURPOSE: To test the ability of a mutant herpes simplex virus (HSV) hrR3 to inhibit growth of Y79 human retinoblastoma in vitro and in vivo. METHODS: Cultured Y79 cells were infected with multiplicities of infection (MOI) ranging from 0.004 to 0.1 of hrR3. Surviving cells were counted using trypan blue dye exclusion. Using X-gal staining, expression of the lacZ gene was examined in vitro on day 3 postinfection to evaluate viral replication. Nude mice harboring Y79 tumors subcutaneously received an intraneoplasmic injection of 5 x 10(7) plaque-forming units of hrR3. The tumor sizes were measured weekly. Expression of the lacZ gene was also examined on one week postinfection. RESULTS: There are 31% and 13% cells surviving in cultured Y79 cells infected by hrR3 at an MOI of 0.1 on days 3 and 5 postinfection respectively compared to those of mock-infected cells. Also more than 70% of Y79 cells were stained with X-gal at an MOI of 0.1 which demonstrated active viral replication in vitro. Virus-treated subcutaneous tumors were smaller than control tumors (p<<0.05, Student's t-test) on days 14, 21, and 28 postinfection. Positive X-gal staining was also observed in the tumor nodule which was challenged with this viral vector. CONCLUSIONS: We have demonstrated that hrR3 is capable of inhibiting Y79 tumor growth both in cell culture and in nude mice. These data suggest that gene therapy using this mutant HSV vector can be a new supplementary therapeutic modality for retinoblastoma.

Animals↗

PubMed1999

Suppression of experimental corneal angiogenesis by focal X-ray irradiation.

PURPOSE: To investigate the effect of focal X-ray irradiation on experimental corneal angiogenesis in the rabbit. METHODS: A gelatin hydrogel sheet impregnated with basic fibroblast growth factor was implanted into the corneal stroma of rabbits; this induced corneal angiogenesis. After the first sign of corneal angiogenesis was noted, the corneal region was irradiated with a dose of 10 Gy or 20 Gy. The control rabbits received no irradiation. The eyes were examined by slitlamp biomicroscopy and photographed, over a period of 28 days. The maximum length and total surface area of corneal angiogenesis were quantified by computerized image analysis. RESULTS: Corneal angiogenesis was noted on day 3 following implantation of the hydrogel sheet. In the rabbits irradiated with 10 Gy, the maximum length and total surface area of corneal angiogenesis were both significantly lower on day 4 and 7 following irradiation, compared to the respective measurement in the controls. In the rabbits irradiated with 20 Gy, the maximum length and total surface area of corneal angiogenesis were significantly lower between days 4 and 21, and between days 4 and 14, respectively, compared to the respective measurement in the controls. CONCLUSIONS: Focal X-ray irradiation to the corneal region suppressed corneal angiogenesis in a dose-dependent manner. Focal X-ray irradiation may be beneficial in treating ocular angiogenesis.

Animals↗

PubMed1999

Relationship between acetazolamide blood concentration and its side effects in glaucomatous patients.

Although acetazolamide, a carbonic anhydrase inhibitor, has an effect of lowering intraocular pressure, a number of side effects have been reported. Therefore, we investigated the relationship between the concentration of acetazolamide and its side effects, including plasma electrolyte imbalance. This study was conducted on 23 glaucomatous patients who received repeated doses of oral acetazolamide for one week or more. The concentrations of total and unbound plasma acetazolamide, as well as in the whole blood from the patients, were measured by high-performance liquid chromatography. The serum creatinine concentration, electrolyte concentrations, and adverse reactions were monitored. We found that plasma concentrations of chloride ion after repeated doses became higher than the normal range. This chloride ion concentration significantly correlated with the acetazolamide concentration in the erythrocytes, but not with the plasma concentration. The patients with erythrocyte acetazolamide concentration more than 20 microg/ml had higher incidences of the side effects. Periodical monitoring of erythrocyte acetazolamide concentration and plasma chloride ion can be easily and safely applied to elderly glaucomatous patients treated with acetazolamide for long periods to prevent overdosage and side effects.

Acetazolamide↗