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Biomedical subjects

T Fukaya

Publications and source records attributed to T Fukaya.

At least 55 records · Page 3Linked to original sources
PubMed1998

The furosemide test and vestibular status in Meniere's disease.

A total of 62 ears of patients with typical Meniere's disease was examined by the furosemide test to detect endolymphatic hydrops. In 95% of the normal control group, the per cent change in the maximum velocity of the slow phase of caloric nystagmus (MVS) after injection of furosemide was under 10%. Therefore, a positive furosemide test was defined as a change in MVS of more than 10%. Thirty-five (56%) of the 62 ears with typical Meniere's disease showed a positive furosemide test. When the affected ears were divided into two groups according to vestibular symptoms, only 11 (38%) of 29 inactive ears were positive while 24 (73%) of 33 active ears were positive. There was a significant difference in the positive rate of the furosemide test between the ears with clinically inactive and active vestibular disease. The per cent canal paresis (CP%) was determined to assess canal excitability and a CP%> 25% was defined as canal paresis. There was no significant difference in the furosemide test positive rate between ears with canal paresis and ears with a normal CP%, although the former tended to show a greater MVS change. The response to the furosemide test showed no relationship to the results of pure tone audiometry. In conclusion, the furosemide test appears to indicate the vestibular status in various stages of Meniere's disease.

Ear Canal↗
PubMed1998

Cyclic changes of vasculature and vascular phenotypes in normal human ovaries.

In order to study alterations of angiogenesis and blood vessel regression through ovarian cycle in human ovaries we quantitatively examined vascularity in various stages in 24 normal human ovaries. Vascular density (VD; vessel numbers/10(-7) m2) and endothelial area of each vessel (EA; 10(-12) m2/vessel) were evaluated using immunohistochemistry of CD34 and CAS 200 image analysis system. Small-sized vessels were sporadically observed in stroma adjacent to primordial or primary follicles (6.73 +/- 1.83 for VD and 113.58 +/- 21.80 for EA). Formation of capillary network was observed in the theca layer of preantral follicles (PA; 15.28 +/- 2.77 for VD and 113.58 +/- 21.80 for EA), and higher density of the capillary network was detected in non-dominant follicles in follicular phase (ND-F) and dominant follicles (DF; 29.33 +/- 3.84 for VD and 179.69 +/- 41.25 for EA). Dense capillary network was still present in non-dominant follicles in luteal phase (ND-L) and atretic follicles (AF; 26.88 +/- 3.36 for VD and 110.88 +/- 50.53 for EA). After ovulation, developing capillaries were also observed in the luteinized granulosa layers in early corpus luteum (21.95 +/- 2.06 for VD and 167.08 +/- 29.59 for EA). Vessel density markedly increased in mid corpus luteum, reached plateau in late corpus luteum (60.85 +/- 5.92 for VD and 70.99 +/- 15.57 for EA) and remained constant during degenerating corpora lutea. Vascular endothelial growth factor was immunohistochemically observed in the theca cells in PA, ND-F, DF and ND-L in follicular stages, and functioning corpora lutea. Immunoreactivity of intercellular adhesion molecule-1 was detected only in post-capillary venules in early degenerating corpora lutea. These findings suggest that ovarian angiogenesis is a requirement for the early stages of folliculogenesis and luteal growth, and also plays an important role in the process of follicular atresia and luteal regression.

Actins↗
PubMed1998

Leukocytes in normal-cycling human ovaries: immunohistochemical distribution and characterization.

We investigated, using an image analysis system, the immunohistochemical localization of leukocyte subpopulations and human leukocyte antigen (HLA)-DR in 30 normal-cycling human ovaries in order to better understand local immunological events in human ovaries. All subtypes of T lymphocytes examined (CD3+, CD4+ and CD8+ cells) were sporadically observed in the stroma and theca layers of follicles throughout the menstrual cycle (ranging from 4.32 to 6.25 cells/10(-7) m2, 1.67 to 3.33 cells/10(-7) m2 and 2.33 to 3.44 cells/10(-7) m2, respectively for the three subtypes), and subsequently, increased in number in atretic follicles (78.70 +/- 6.90, 31.13 +/- 2.54 and 43.31 +/- 3.35). After ovulation, the number of T lymphocytes was markedly low in the early and mid corpus luteum (13.88 +/- 1.62, 4.18 +/- 0.50 and 6.53 +/- 0.45). The number increased in the late corpus luteum, and was highest in the late degenerating corpus luteum (255.67 +/- 27.10, 102.12 +/- 7.80 and 137.34 +/- 12.50). HLA-DR was sporadically positive in fibroblasts in the stroma and theca layers of follicles (means ranged from 1.25 to 1.82 cells/10(-7) m2), and increased in atretic follicles (24.68 +/- 2.25). HLA-DR+ cells were markedly low in the early and mid corpus luteum (2.16 +/- 0.88), increased in the late corpus luteum, and reached a plateau in the late degenerating corpus luteum (121.84 +/- 17.73). The great majority of these increased HLA-DR+ cells were macrophages. Results of our study suggest that T lymphocytes and/or macrophages play important roles in luteal regression and follicular atresia in normal-cycling human ovaries.

Adult↗
PubMed1998

Increased mast cell density in peritoneal endometriosis compared with eutopic endometrium with endometriosis.

PROBLEM: Factors regulating fibrosis in peritoneal endometriosis are poorly understood. We hypothesized that mast cell proteases may play a role in the process of fibrosis in peritoneal endometriosis. As the first step in examining the possible contribution of mast cells to fibrosis, we investigated their distribution in peritoneal endometriosis lesions according to their macroscopic appearance. METHOD OF STUDY: Mast cells were identified by immunohistochemistry using monoclonal antibody against mast cell tryptase on formalin-fixed, paraffin-embedded sections. Mast cell density (mean mast cell count per 0.13 mm2 of stroma) was measured using a computerized image analysis system. RESULTS: Mast cell density was significantly increased in black peritoneal lesions compared with red peritoneal lesions in endometriosis. CONCLUSIONS: Mast cells may be involved in the pathogenesis of peritoneal endometriosis.

Chymases↗
PubMed1998

Growth hormone improves mouse embryo development in vitro, and the effect is neutralized by growth hormone receptor antibody.

We investigated whether growth hormone(GH) may influence the development of mouse embryo in vitro using recombinant GH and anti-growth hormone receptor(GHR) antibody. When mouse two-cell-stage embryos were cultured with GH, 5, 10 mIU/ml, the rates of formation of both blastocysts and of hatched blastocysts from two-cell-stage embryos were significantly higher than in the medium without GH. The number of blastomeres from blastocyst treated with GH, 2, 5 and 10 mIU/ml, was significantly higher than those cultured without GH. When cultured blastocysts were implanted in recipient uteri, a significantly higher implantation rate was found for the GH-treated(10 mIU/ml) mouse embryos than for controls. To confirm the GH effect, anti-GHR monoclonal antibody MAb5 was employed to neutralize the action of GH through GHR. Two-cell-stage mouse embryos were cultured with GH, 10 mIU/ml, and anti-GHR antibody, and as a control, embryos were cultured with GH alone. Both the rate of formation of blastocysts from two-cell-stage embryos and the number of blastomeres were significantly decreased in groups exposed to anti-GHR antibody. Results indicate that GH may be important to early embryonic development, acting through GHR. GH may help to obtain good quality embryos and improve the implantation rate in IVF programs.

Animals↗
PubMed1998

Analysis of clonality in human endometriotic cysts based on evaluation of X chromosome inactivation in archival formalin-fixed, paraffin-embedded tissue.

The clonality of epithelial cells in ovarian endometriotic cysts was evaluated on the basis of a polymorphism of the X-linked phosphoglycerate kinase gene (PGK). The problems associated with clonality analysis of DNA samples extracted from archival formalin-fixed, paraffin-embedded tissue were addressed with the use of newly designed primers and a modified stepdown program for PCR amplification. The method relies on digestion of DNA with the methylation-sensitive restriction enzyme SnaBI, PCR amplification of PGK, and detection of a BstXI polymorphism. With this approach, only the inactive (methylated) PGK allele is selectively amplified by PCR. A total of 25 epithelial cells in endometriotic cysts and 25 matched normal ovarian stromal tissue specimens were analyzed. All of the control tissue samples yielded PCR products, with 11 of the 25 samples found to be heterozygous for the BstXI polymorphism. Ten of the 25 epithelial cells in endometriosis specimens were informative at this locus, and all of these 10 were shown to be monoclonal on the basis of PGK methylation. The results indicate that this new method of clonal analysis with archival formalin-fixed tissue is reliable and confirm that endometriotic cysts are monoclonal in origin.

Base Sequence↗
PubMed1997

Increased secretion of hepatocyte growth factor by eutopic endometrial stromal cells in women with endometriosis.

OBJECTIVE: We evaluated the secretion of hepatocyte growth factor by eutopic endometrial stromal cells in vitro. DESIGN: Eutopic endometrial stromal cells were isolated and the culture supernatants were collected after 48 hours of incubation. The secretion of hepatocyte growth factor was analyzed by ELISA. SETTING: Department of Obstetrics and Gynecology of Tohoku University Hospital. PATIENT(S): Specimens of endometrium in the proliferative phase were excised from 11 patients who were undergoing laparoscopy for infertility. Six of the patients had endometriosis and five did not. None of the patients had received hormonal treatment before surgery. RESULT(S): Hepatocyte growth factor secretion was found to be increased significantly in cultured endometrial stromal cells from the infertile patients with endometriosis compared with those without endometriosis. CONCLUSION(S): The secretion of hepatocyte growth factor was up-regulated in eutopic endometrial stromal cells from patients with endometriosis. An increase in hepatocyte growth factor production may be characteristic of endometriosis and may be involved in the pathophysiology of this disease.

Adult↗
PubMed1997

Extracranial meningioma presenting as a tumour of the external auditory meatus: a case report.

A rare case of extracranial meningioma presenting as a tumour of the external auditory meatus is reported. Biopsy indicated a diagnosis of meningioma, but the radiological appearance was unusual. For example, computed tomography (CT) scans showed an unenhanced tumour mainly located in the squamous part of the temporal bone which expanded into the external meatus destroying the temporal bone. Magnetic resonance imaging (MRI) revealed that the tumour did not extend into the intradural space. This meningioma, had an obvious tendency for extracranial development. According to the operative findings, the tumour arose from the middle cranial fossa dura and extended through the air cells of the temporal bone into the external meatus, instead of growing intracranially. Secondary extracranial meningiomas of the temporal bone usually have a large intracranial component and cause neurological symptoms. However, this was a very rare case of a small meningioma causing no symptoms except for conductive hearing loss.

Adult↗
PubMed1997

Macrophages in normal cycling human ovaries; immunohistochemical localization and characterization.

We evaluated the immunolocalization and characterization of macrophages in 28 normal cycling human ovaries. Two primary antibodies were used to detect the macrophages: PGM1, a general marker for macrophages, and 25F9 which is specific for phagocytosing macrophages. Spindle-shaped cells positive for PGM1 but negative for 25F9 were observed in the stroma (123.6 +/- 1.05 cells/10(-6) m2) and theca layer of the follicle (mean ranged from 22.61 to 53.79) and the number of these cells did not change throughout the cycle. After ovulation, PGM1 positive cells with ballooning bodies began to appear in the early corpus luteum (111.8 +/- 0.83). The number of these macrophages increased in the mid and late corpora lutea, and reached maximum in the early degenerating corpus luteum (1231.0 +/- 3.29). A lower number of PGM1 positive ballooning macrophages were observed in the atretic follicle (177.9 +/- 1.42). 25F9 positive cells were also observed among the PGM1 positive balloon-shaped cells. The number of cells double positive for 25F9 and PGM1 was observed in the mid corpus luteum (44.6 +/- 0.46), increased in the late corpus luteum and early degenerating corpus luteum, and reached plateau in the late degenerating corpus luteum (549.0 +/- 5.82). A lower number of these double positive macrophages were also observed in the atretic follicle (64.8 +/- 0.36). The ratio of 25F9 to PGM1 positive cells increased in parallel with ageing of the corpus luteum (0.19 in the mid corpus luteum, 0.39 in the late corpus luteum, and 0.37 in the early degenerating corpus luteum), and the great majority of PGM1 positive cells were also immunopositive for 25F9 in the late degenerating corpus luteum (0.81). These results suggest that in normal cycling human ovaries, macrophages are mainly involved in luteal regression as scavengers.

Antibodies, Monoclonal↗
PubMed1997

Hepatocyte growth factor stimulated proliferation, migration, and lumen formation of human endometrial epithelial cells in vitro.

Human endometrial regeneration has been considered to be modulated by several growth factors. However, little is known about the detailed mechanisms involved in the repair of the endometrium during the menstrual period. Hepatocyte growth factor (HGF) is a pleiotropic growth factor that reportedly acts on various epithelial cells. In the present study, we observed HGF and mRNA expression in human endometrium and mRNA expression in cultured endometrial epithelial cells of the c-met gene by reverse transcription-polymerase chain reaction and Southern blot hybridization. We also examined the biological role of HGF on the regeneration of the endometrium by using cultured endometrial epithelial cells in their proliferative phase. With the proliferation assay, the addition of 10-50 ng/ml of HGF showed that HGF was mitogenic in a dose-dependent manner. With Boyden's chamber technique, 50 ng/ml of HGF significantly stimulated cell migration. In a three-dimensional cell-culture system, the endometrial epithelial cells formed cell clusters and gradually formed epithelial lumens, both of which were stimulated by 50 ng/ml of HGF. Results suggest the HGF stimulates the proliferation and migration of, and morphogenic changes in, endometrial epithelial cells. HGF may modulate the regeneration of the endometrium during menstruation.

Adult↗
PubMed1997

Does apoptosis contribute follicular atresia and luteal regression in human ovary?

We investigated apoptosis in 37 human ovaries from normally cycling women to determine the mechanism of the various stages of follicular atresia and the regression of corpus luteum in human. Apoptosis was examined by 3'-hydroxy nick end-labeling, by immunostaining of an apoptosis-related antigen, Le(y) and by cell death-related genes, bcl-2. Immunoreactivity for Le(y) and nick end-labeling reactive cells were not observed in the follicular and luteal phases, except for scattered cells in the degenerating corpus luteum. On the other hand, immunoreactivity for bcl-2 was observed in the granulosa cells of antral follicles. These results indicated that apoptosis is not obvious through follicular maturation and luteal regression. This may be due to the relatively long process of human follicular growth and atresia.

Adult↗
PubMed1997

Keratinocyte growth factor and receptor mRNA expression in cholesteatoma of the middle ear.

Keratinocyte growth factor (KGF) is synthesized and secreted exclusively by stromal cells in epithelialized organs, and specifically promotes proliferation of cells of epithelial origin, including keratinocytes. Cholesteatoma is a proliferative form of keratinocyte dysregulation with aggressive growth that often leads to destruction of adjacent bone. We examined the expression of KGF, closely related peptides and their receptors in the development of cholesteatoma by comparing cholesteatoma with normal ear skin using the reverse transcription-polymerase chain reaction (RT-PCR) followed by Southern blot analysis. All surgical specimens of cholesteatoma and most normal ear skin samples expressed detectable levels of KGF and KGF receptor (KGFR). Semiquantitative RT-PCR using beta-actin mRNA as an internal standard revealed significantly higher expression of KGF mRNA in cholesteatoma than in normal skin, while no significant difference in KGFR mRNA expression was noted between cholesteatoma tissue and normal skin. These results suggest that excessive KGF synthesis may contribute to the hyperproliferative state in cholesteatoma.

Blotting, Southern↗
PubMed1997

[The impact of conjugal bereavement and the buffering effect of social support on the health of elderly people].

This study examined the impact of the spouse's death on the mental and physical health of the elderly, sixty years and older, and the buffering effect of social support against the impact. A three-year study was conducted of 1,087 people whose spouses were alive at the time of the initial survey. Changes over the three-year period were compared among the following three groups: (1) the spouse died within a year prior to the second survey (N = 21); (2) the spouse died more than a year before the survey (N = 47); and (3) the spouse was still alive (N = 901: the comparison group). Results were as follows: (1) Mental and physical health declined more rapidly in the first group than the comparison group, while no significant change was found for the second group. (2) Social support after the spouse loss significantly helped buffer the negative effect on the mental health, but support prior to the loss had no such effect. Social support had no moderating effect on the physical health.

Aged↗
PubMed1996

Expression of growth hormone receptor in mouse preimplantation embryos.

Growth hormone (GH) is believed to be involved in reproductive function. Recent reports demonstrate the presence of the growth hormone receptors (GHR) in reproductive organs and fetal tissues suggesting that the contribution of GH to reproductive phenomena may be mediated through GHR. In this study we assessed the expression of GHR in mouse preimplantation embryos using immunofluorescent staining with a monoclonal antibody directed against GHR. The messenger RNA (mRNA) for GHR was investigated in mouse preimplantation embryo by reverse transcription-polymerase chain reaction (RT-PCR). The anti-GHR monoclonal antibody Mab 263 was detected by immunofluorescent staining after the compaction of the morula stage. GHR mRNA transcript was identified in 8-cell stage embryos. Our findings indicate that GHR is localized in mouse preimplantation embryos, and suggests that GH has a direct effect on preimplantation embryos mediated by GHR.

Animals↗
PubMed1996

Immunohistochemical study of steroidogenesis and cell proliferation in polycystic ovarian syndrome.

We evaluated the immunolocalization of the steroidogenic enzymes involved in the production of ovarian steroids, including the cholesterol side-chain cleavage enzyme (P450scc), 3beta-hydroxysteroid dehydrogenase (HSD), 17alpha-hydroxylase (P450c17) and aromatase (P450arom), oestrogen receptor (ER) and androgen receptor (AR), a steroidogenic transcription factor. Ad4-binding protein (Ad4BP) and a cell cycle-related nuclear antigen, Ki67, in five patients with polycystic ovarian syndrome (PCOS). Results were compared with those from normal cycling human ovaries to study in situ ovarian steroidogenesis and cell proliferation in polycystic ovaries (PCO). We classifed the follicles morphologically according to the development of granulosa types: type A, more than four layers (n = 7); type B, one to three layers (n = 11); and type C, theca interna cells only (n = 21). ER and P450arom were not observed in any of the follicles examined. In type A follicles, P450scc, 3beta-HSD, P450c17, AR and Ad4BP were observed in theca cells in all seven follicles examined, but the granulosa cells were positive only for Ad4BP (4/7) and AR (7/7). These immunohistolocalization patterns resembled those in non-selected antral follicles of normally cycling human ovaries. In theca cells from types B and C follicles, follicles positive for the steroidogenic enzymes, AR and Ad4BP were decreased in number. There were no significant differences between types A and B PCO follicles in the Ki67 labelling index of granulosa or theca cells, and between PCO and antral follicles from normally cycling human ovaries. Data demonstrate that the follicles of PCO are by no means atretic and are actively involved in both steroidogenesis and cell proliferation. The absence of ER and aromatase expression in the granulosa cells of PCO may be important in abnormal follicular development in patients with PCOS.

3-Hydroxysteroid Dehydrogenases↗
PubMed1996

Trisomy 18 mosaicism associated with secondary amenorrhea: ratios of mosaicism in different samples and complications.

A 28-year-old woman who complained of irregular menstruation was diagnosed as suffering from trisomy 18 mosaicism. She was karyotyped because of her characteristic face, mild mental retardation and aberrant hyperpigmentation of the skin. Her motor function was within normal range. Physical and laboratory examinations, however, revealed obesity, short stature, minor anomalies of the fingers, many areas of hyperpigmentation on the trunk and the hips, hypergonadotropinemia, diabetes mellitus, liver dysfunction, and hyperlipidemia. The ratios of normal/trisomy 18 were 4:135 in blood lymphocytes, 3:11 in a hyperpigmented area of the skin, 20:0 in a normally pigmented area of the skin, and 14:6 in ascitic cells. Laparoscopy revealed that her ovaries contained neither follicles nor germ cells.

Abnormalities, Multiple↗
PubMed1996

Mutation of cytochrome P-45017 alpha gene (CYP17) in a Japanese patient previously reported as having glucocorticoid-responsive hyperaldosteronism: with a review of Japanese patients with mutations of CYP17.

A 17-yr-old female Japanese patient, who was reported in 1968 as having glucocorticoid-responsive hyperaldosteronism but was presumed to have a defect of 17 alpha-hydroxylation mainly in the adrenal glands as the etiology of her illness, was followed. The relationship between clinical manifestations and molecular abnormalities in cytochrome P-45017 alpha gene (CYP17) was also reviewed based on the literature on Japanese patients with 17 alpha-hydroxylase deficiency. She has been treated with dexamethasone, resulting in normal blood pressure and normokalemia for 28 yr. She had almost normal gonadal function with regular menstruation on her first admission. Because of sustained genital bleeding, however, she underwent total hysterectomy with an ovarian biopsy at the age of 42 yr. No follicles or corpus luteum were detected in the ovarian specimen. At the age of 45 yr, the basal levels of sex steroids were decreased, while those of gonadotropins were increased. A genetic study on CYP17 revealed a homozygous deletion of phenylalanine (Phe) codon (TTC) at either amino acid position 53 or 54 in exon 1. A review of the literature revealed 4 patients with this type of CYP17 mutation, including the present patient, out of a total of 11 young adult Japanese patients. The clinical manifestations caused by congenitally deficient gonadal function were not marked in any of these 4 patients, but were marked in 5 of the 7 patients with different mutations of CYP17. The remaining 2 female patients had irregular menstruation. The pretreatment urine/plasma values of aldosterone were variable, normal to high, in individual patients, regardless of the structural abnormalities of CYP17. The following conclusions were suggested: 1) this type of CYP17 mutation is associated with well preserved gonadal function in young adult patients, but it likely causes early reduction of gonadal function with increasing age in these patients; 2) the prevalence of this type of CYP17 mutation is quite high in Japanese patients; and 3) the pretreatment hyperaldosteronism observed in the present patient seems not to be related to the mutation of CYP17.

Adolescent↗