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Biomedical subjects

T Fukaya

Publications and source records attributed to T Fukaya.

At least 73 records · Page 4Linked to original sources
PubMed1996

Cell turnover in normal cycling human ovary.

We investigated cell proliferation and apoptosis in 37 normal cycling human ovaries to determine cell turnover in the various stages of follicular and luteal development. We examined cell proliferation by immunostaining for both Ki67 and proliferative cell nuclear antigen (PCNA) and by silver staining of nucleolar organizer regions (AgNORs). Apoptosis was examined by 3'-hydroxy nick-end labeling and by immunostaining of an apoptosis-related antigen, Ley. The labeling indexes of Ki67, PCNA, and AgNORs were significantly increased in antral follicles. However, there were no significant differences in the labeling index of Ki67, PCNA, and AgNORs between dominant and nondominant follicles, including nonovulated follicles in the luteal phase. These results indicate that the transformation of granulosa cells from quiescence to active growth is important in early folliculogenesis. Immunoreactivity for Ki67 and PCNA were observed predominantly in the functioning corpus luteum, but not in the degenerating corpus luteum, indicating proliferation only during the luteal phase. Immunoreactivity for Ley and nick end-labeling reactive cells were not observed in the follicular and luteal phases, except for scattered cells in the degenerating corpus luteum. This may be because of the relatively long process of human follicular growth and atresia.

Apoptosis↗
PubMed1996

Indications for laparoscopic surgery of ovarian tumors.

To investigate the criteria for laparoscopic surgery in ovarian tumors, a retrospective review of indications, preoperative assessments using transvaginal ultrasonography, serum CA 125, surgical methods and diagnosis and post-operative complications were analyzed. Forty-three patients underwent surgical treatment for ovarian tumors, from November 1992 to October 1993. Of the 43 patients with ovarian tumors, 19 (44%) underwent laparoscopic surgery, and 24 (56%) underwent laparotomy. Laparoscopic surgery was performed in 18 of 34 (53%) patients with benign ovarian cysts. Eight patients with malignant ovarian tumors were treated by laparotomy. Coincidence of preoperative assessment and surgical diagnosis was 95%. No major complications were observed in the laparoscopic surgery. We suggest that malignant ovarian tumors and inextricable adhesions are contraindications to laparoscopic surgery. We propose the following criteria for laparoscopic surgery by classification into three groups: A) good indication: benign cystic tumors with good mobility, B) deliberation indication: benign tumors with poor mobility, including some dermoid cysts and endometrial cysts, C) contraindication: malignant tumors and dense adhesions. We conclude that indications of laparoscopic surgery for ovarian tumors should be determined by careful preoperative assessment using transvaginal ultrasonography and tumor markers.

CA-125 Antigen↗
PubMed1996

Juvenile adenomyotic cyst of the corpus uteri with dysmenorrhea.

The clinical and pathological features of an apparently unique case of an endometrial cyst of the uterus are reported. The cyst was located within the myometrium of a 16-year-old woman suffering from dysmenorrhea. After excision of the cyst, patient's symptoms improved. On histological examination, the cyst most closely resembled an adenomyotic cyst.

Adolescent↗
PubMed1996

Is direct cell-to-cell contact needed to improve embryonic development in co-culture?

In vitro co-culture of embryos and somatic cells is used to obtain well-developed embryos of humans and other species. However, it is not known whether direct cell-to-cell contact is essential to embryonic development. To elucidate this mechanism, we cultured mouse embryos using rabbit oviductal cell as a somatic cell. To avoid the direct contact we used a microporous membrane cell-culture insert. This cell-culture insert permits only the liquid portion of the culture medium to pass through it and was interposed between mouse embryos and cultured rabbit oviductal cells. When mouse embryos were placed on the cultured oviductal cells directly, 52.2% of two-cell-stage embryos developed to the blastocyst stage. In contrast, when the cell-culture insert was interposed between the embryos and the cultured oviductal cells, 50.0% of two-cell-stage embryos developed to that stage. There was no difference between the rate of blastocyst development in co-culture system with and without cell culture insert. Results indicate that oviductal factor (s), rather than a direct contact with oviductal cells, is essential to the enhancement of embryonic development in vitro.

Animals↗
PubMed1996

[Case report of one pediatric patient surviving 6 years after heart transplantation in the U.K].

In October 1988, a six year old Japanese girl with restrictive cardiomyopathy underwent heart transplantation (HTx) at Harefield Hospital in the U.K. Six years 9 months after her HTx, she is doing well and is the longest survivor among the Japanese HTx recipients. Cardiac catheterization 5 years after her HTx showed good cardiac function with LVEF of 54.7%. Coronary angiogram at that time showed no abnormal findings. Cardiac biopsy revealed no sign of rejection or myocyte degeneration. LVDd (Left Ventricular Diastolic Dimension) grew from 35.5 mm at the time of transplant to 42.6 mm 5 years later by echocardiogram, suggesting the growth of the transplanted heart. Her good progress, including catch-up growth and good QOL, reflects the excellent benefit of HTx today. The authors deeply appreciate the courtesy of Harefield Hospital in accepting to perform her HTx.

Adolescent↗
PubMed1995

Localization of microfilaments during oocyte maturation of golden hamster.

The localization and changes in microfilaments (MF) during golden hamster oocyte maturation were examined by an immunofluorescein method and confocal laser scanning microscopy (CLSM). We also studied the relationship between the changes in MF and oocyte nuclear and cytoplasmic maturation. During in vivo maturation, generalized submembranous MF were found initially which gradually became more prominent at the site of the first polar body extrusion. However, 43.7% of the in vitro matured metaphase 2 stage oocytes lacked the submembranous MF structure. This fact may partly account for the low fertilization rate of in vitro matured oocytes. MF were not found in the folicular oocytes cultured in cytochalasin D-containing medium, and metaphase-like chromosomes were located at the center of the oocyte and first polar body extrusion did not occur. Twenty-five percent of the oocytes, which were arrested at meiosis by hypoxanthine, synthesized submembranous MF structure although the nuclear stage of these oocytes was germinal vesicle. These facts suggest that MF plays a role in nuclear behavior but there are some differences in the changes taking place within the nucleus and MF. MF may play a role in oocyte cytoplasmic maturation although the details of this have yet to be established.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1995

Laser vaporization of the ovarian surface in polycystic ovary disease results in reduced ovarian hyperstimulation and improved pregnancy rates.

OBJECTIVE: Our purpose was to examine the efficacy of laser vaporization of the ovarian surface in polycystic ovary disease to reduce repeated ovarian hyperstimulation syndrome and thereby improve pregnancy outcome. STUDY DESIGN: Twenty-six infertile patients with polycystic ovary disease who previously had ovarian hyperstimulation syndrome after stimulation with human menopausal gonadotropin and who failed to conceive were studied. All patients were treated by potassium titanyl phosphate and neodymium-yttrium-aluminum-garnet laser and evaluated. Patients not ovulating spontaneously after vaporization were treated with either clomiphene citrate or human menopausal gonadotropin. RESULTS: After vaporization spontaneous ovulation was confirmed in six patients. For ovulation induction three patients received clomiphene citrate and 17 received human menopausal gonadotropin. Of the patients treated with human menopausal gonadotropin, mild ovarian hyperstimulation syndrome was found in three patients, and the incidence of ovarian hyperstimulation syndrome decreased significantly. Pregnancy was confirmed in 19 of 26 patients. CONCLUSION: Laser vaporization is promising for the prevention of ovarian hyperstimulation syndrome and improving pregnancy outcome in patients with polycystic ovary disease who have previously had ovarian hyperstimulation syndrome.

Adult↗
PubMed1995

Transcription factor adrenal 4 binding protein as a marker of adrenocortical malignancy.

Adrenal 4 binding protein (Ad4BP) is a transcription factor that regulates the expression of the steroidogenic enzymes and is expressed primarily in steroidogenic cells. We immunolocalized Ad4BP in adrenocortical carcinoma (eight cases) and various malignancies that histologically simulate an adrenocortical carcinoma to evaluate the value of Ad4BP as an immunohistochemical marker of adrenocortical carcinoma. These malignancies examined were renal cell carcinoma (20 cases), hepatocellular carcinoma (10 cases), malignant melanoma (eight cases), ovarian (six cases) and uterine (three cases) clear cell carcinoma, large cell carcinoma of the lung (five cases), and pheochromocytoma (three cases). Nuclear Ad4BP immunoreactivity was observed only in adrenocortical carcinoma cases but not in other tumors examined. Almost all of the adrenocortical carcinoma cells were immunohistochemically positive for Ad4BP including cells associated with bizarre nuclei. These results show that application of Ad4BP immunostain can contribute greatly to the differential diagnosis of adrenocortical carcinoma.

Adrenal Cortex Neoplasms↗
PubMed1995

Expression of epidermal growth factors and epidermal growth factor receptor in normal cycling human ovaries.

Immunolocalization of transforming growth factor-alpha (TGF alpha), epidermal growth factor (EGF), cripto-1, amphiregulin and epidermal growth factor receptor (EGFR) was studied in 51 premenopausal human ovaries at various phases of the menstrual cycle. Localization of mRNA for TGF alpha and EGF was also studied by in-situ hybridization. Immunoreactive TGF alpha was observed predominantly in theca cells in 12 of 33 antral follicles in the follicular phase (6/14 dominant follicles, and 6/19 non-dominant) but not in any of the 18 follicles in the luteal phase or in primordial and pre-antral follicles. TGF alpha immunoreactivity was present predominantly in the luteinized granulosa cells in 13 of 15 corpora lutea in the luteal phase, which are considered to be active in steroidogenesis, but not in any of the regressed corpora lutea. Accumulation of TGF alpha mRNA hybridization signal was observed only in the theca cells in the follicles and luteinized theca cells in the ovaries that were immunohistochemically positive for TGF alpha. EGFR immunoreactivity was detected in 24 of 33 antral follicles in the follicular phase and in two of 18 follicles in the luteal phase but not in any of the corpora lutea. Immunoreactive EGF, cripto-1 and amphiregulin or EGF mRNA was not detected in any follicles, corpora lutea, or the stroma cells examined. These results indicate that, of the epidermal growth factors examined in this study, TGF alpha is locally synthesized in normal cycling human ovaries and TGF alpha may be synthesized in theca cells and act on the granulosa cells in a paracrine fashion through the EGFR in ovarian follicles.

Adult↗
PubMed1995

Ad4BP in the human adrenal cortex and its disorders.

Ad4BP, a zinc finger DNA-binding protein, is a transcription factor that regulates the expression of the steroidogenic P450 genes. We performed immunoblotting and immunohistochemistry of Ad4BP in 34 human adrenal cortex specimens, which included adrenocortical adenomas and carcinomas. Immunoblotting revealed a single band of 53K, corresponding to the mol wt of Ad4BP. The immunohistochemical studies demonstrated that Ad4BP immunoreactivity was present exclusively in the nuclei of nearly all of the adrenocortical parenchymal cells in both the normal and the pathological human adrenal specimens. Ad4BP was immunostained with equal intensity and frequency among the different cell types. Ad4BP immunoreactivity was also observed in areas of marked degenerative changes, such as lipomyelomatous lesions, and in poorly differentiated carcinoma cells. These results suggest a close association of Ad4BP expression with the biological phenotype of adrenocortical parenchymal cells. Ad4BP therefore seems to play important roles in the induction and maintenance of the transcription of all steroidogenic P450 genes in human adrenocortical cells, even after malignant transformation.

Adenoma↗
PubMed1995

Immunohistochemical localization of Ad4-binding protein with correlation to steroidogenic enzyme expression in cycling human ovaries and sex cord stromal tumors.

Ad4-binding protein (Ad4BP) has been demonstrated recently as a transcription factor that serves as a general regulator of all steroidogenic P450 genes. We examined the expression of Ad4BP in 32 normal cycling human ovaries and 22 human ovarian sex cord stromal tumors by immunoblotting and immunohistochemistry. Immunoblotting of normal cycling human ovaries revealed a single band of 53 kilodaltons, corresponding to the mol wt of Ad4BP. We also correlated Ad4BP expression with the immunolocalization of the steroidogenic enzymes (side-chain cleavage cytochrome P450, cytochrome P450 17 alpha-hydroxylase, and cytochrome P450 aromatase). Ad4BP immunoreactivity, which was present only in the nuclei, was observed sporadically in the granulosa cells and adjacent stromal cells in the preantral follicles. In the dominant antral follicles, Ad4BP was detected in both granulosa and theca interna cells. However, in the nondominant antral follicles, Ad4BP was observed only in theca interna cells. In the corpus luteum, Ad4BP was present in both luteinized granulosa and thecal cells. Ad4BP was also expressed in some atretic follicles and degenerating corpora lutea. The spatial and temporal localization of Ad4BP in the normal cycling human ovary generally correlated well with that of steroidogenic enzymes. However, expression of the steroidogenic enzymes followed that of Ad4BP during the developing stages of the preantral follicle and vice versa during the process of follicular atresia. In ovarian sex cord stromal tumors, Ad4BP expression was observed in tumor cells that were positive for steroidogenic enzymes, but not in nonsteroidogenic tumor cells. These results, especially the in situ colocalization of Ad4BP and the steroidogenic enzymes, suggest that Ad4BP has the potential to control steroidogenic P450 expression in both normal and pathological human ovaries.

Adult↗
PubMed1995

Sperm motility characteristics and pregnancy outcome of artificial insemination with husband's semen for male infertility.

What parameter of semen has influence on the pregnancy outcome of artificial insemination with the husband's semen (AIH) with washed sperm for male infertility was investigated. Two hundred fifteen cycles in 62 patients of AIH with washed sperm for male infertility were prospectively studied. We compared six parameters (total sperm count, motility rate, sperm motile efficiency (SME) both before and after washing) in pregnant and non-pregnant cycles. SME of washed sperm was significantly higher in the pregnant than in the non-pregnant cycles (241.0 +/- 68.0 vs. 169.5 +/- 80.0). There were no differences in other five parameters between pregnant and non pregnant cycles. SME, the parameter of sperm motility characteristics, of washed sperm is closely related to the outcome of AIH for male infertility. Sperm motility characteristics after washing may be one of the important indexes of the result of AIH for male infertility.

Adult↗
PubMed1995

[Evaluation of the furosemide test using the air caloric stimulator].

Both the furosemide test, and the glycerol test have been reported as being effective for detection of endolymphatic hydrops. In the furosemide test, the caloric test is performed first, and then repeated 40 minutes after i.v. injection of furosemide, 20 mg. The maximum velocities of each caloric nystagmus are compared, so the caloric stimulus conditions must be constant. In the original method, the caloric test is performed by irrigation with 50 ml of water at 30 degrees C or 44 degrees C for 20 seconds. Since it is difficult to accurately maintain the water temperature at a constant level, however, we use the air caloric stimulator NCA-105 (ICS) for the furosemide test. By using this stimulator, we can always perform the caloric test under the same conditions. The conditions of air irrigation in 28 normal subjects were set at 37 +/- 11 degrees C, 61/min, and 60 seconds. These conditions were confirmed in other normal subjects to fairly well correspond to water irrigation (30 degrees C or 44 degrees C, 50 ml, 20 seconds). Next we reassessed the furosemide test by using the air caloric stimulator in normal subjects. As a result, positive effects were observed in 7.4% of the normal subjects. This finding was similar to that reported in the literature (Futaki et al., 1971). The air caloric test is considered to be a more useful examination than the water caloric test because it is less unpleasant for the subject and easily provides the same conditions before and after furosemide administration.

Adolescent↗
PubMed1995

[Human folliculogenesis and local regulation].

It is well known that gonadotropin controls a major part of follicular development. However, the mechanism of local regulation under the control of gonadotropins in still unclear. In this study, we focused on the local regulation of steroidogenesis, growth factors and cell proliferation to evaluate the human follicular development. To assess steroidogenesis, it is important to detect the expression of steroidogenic enzymes in the granulosa and theca cells during folliculogenesis. We initially tried to find out the transcription factor Ad4BP that binds the Ad4 site and regulates the function of steroidogenic enzyme. By immunohistochemistry, the expression of Ad4BP was confirmed sporaf1p4lly in preantral granulosa cells. In the antral follicles, the expression of Ad4BP was observed both in the granulosa and theca cell. According steroidogenic enzyme, we evaluated temporal and spatial localization of cholesterol side chain cleavage (scc), 3 beta hydroxysteroid dehydrogenase (3 beta HSD), 17 alpha hydroxylase (17 alpha) and aromatase, and steroid receptors. In briefly, the localizations of scc, 3 beta HSD and 17 alpha were observed in preantral follicles and the mRNA expressions of these enzymes were confirmed in the theca cell by in situ hybridization method. Expression of aromatase was generally observed in only one follicle (antral or mature follicle) per case in mid proliferative to premenstrual phase. The localization of androgen and estrogen receptor was observed in the antral follicle granulosa cells, and estrogen receptor was detected only in aromatase positive follicles. These results suggested that Ad4BP initially controls the function of steroidogenic enzymes and steroidogenic enzymes gradually express from primary follicles to mature follicles. At antral follicle stage, steroid metabolism completes to produce testosterone. When aromatase and estrogen receptor express in antral follicle, this antral follicle develops as the dominant follicle and produces estradiol to promote follicle maturation. We therefore speculate that the expression of aromatase and estrogen receptor have an important role for the selection of dominant follicle in human. According growth factors for follicular development, it has been demonstrated to be important in the biological activity in the ovary. In this study, we examined the localization of EGF, TGF alpha and their receptor (EGFR). The localization of EGF was not confirmed both mRNA and protein level through follicular development. On the other hand, the localization and expression of TGF alpha was confirmed in theca cells and EGFR in granulosa cells at antral stage.(ABSTRACT TRUNCATED AT 400 WORDS)

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

Bilirubin helps to overcome the two-cell block in mouse oocyte cultures.

PURPOSE: In vitro fertilization and culture of mouse oocytes, under normal atmospheric oxygen tension, subjects them to severe oxidative stress. Oocytes from some strains of mice lack the natural protective mechanism that guards them against this oxidative stress and fail to develop beyond the two-cell stage. METHODS: We could overcome the toxic effects of oxygen metabolites by adding 0.2-0.4 mg/dl bilirubin in a lactate-pyruvate culture medium defined by Whitten (1971). Six- to 8-week-old ICR (Institute of Cancer Research) female mice were super ovulated by intra peritoneal injection of 5 IU PMSG (pregnant mare serum gonadotropin) followed by 10 IU hCG 48 h later. The oocytes were collected from the distended fallopian tubes and inseminated with 1-2 million sperm from 3-4-month-old ICR male mice. The eggs were scored at 24, 48, and 72 h after the hCG injection. CONCLUSIONS: With 0.4 mg/dl bilirubin supplement, by the end of 72 h, 82% of the eggs progressed from the two-cell stage to the four-cell stage. Routine inclusion of bilirubin can improve embryo development in vitro.

Animals↗
PubMed1994

Immunohistochemical localization of growth hormone receptor in cyclic human ovaries.

It has been demonstrated that co-administration of growth hormone (GH) augments ovarian response to human menopausal gonadotrophin, and GH itself increases steroidogenesis in cultured ovarian cells. It is not clear, however, whether the effects of GH on the ovary are direct or not. We performed immunohistochemistry using specific monoclonal antibodies against human GH receptor on 51 specimens of pre-menopausal human ovaries from various phases of the menstrual cycle to detect and localize GH receptors. Immunohistochemical localization of P450 aromatase and 3 beta-hydroxysteroid dehydrogenase was performed using these enzymes as immunohistochemical markers for steroidogenesis in these ovaries. GH receptor immunoreactivity was observed in luteinized granulosa cells in corpora lutea in the luteal phase, which are considered to be active in steroid production. In the follicular phase, GH receptor immunoreactivity was detected in the granulosa layer of only three out of 35 antral follicles. These results demonstrate that immunoreactivity of GH receptor is present in human ovaries, suggesting a direct action of GH on human ovarian functions, especially during luteal phase.

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

Immunohistochemical distribution of progesterone, androgen and oestrogen receptors in the human ovary during the menstrual cycle: relationship to expression of steroidogenic enzymes.

In order to characterize immunohistochemically the possible in-situ effects of gonadal steroid hormones in the human ovary during the menstrual cycle, we immunolocalized progesterone (PR), androgen (AR) and oestrogen (ER) receptors in 50 normal cycling human ovaries, and examined the relationship between these findings and the cellular localization of steroidogenic enzymes including cytochrome P-450 cholesterol side-chain cleavage (P-450scc) enzyme, 3 beta-hydroxysteroid dehydrogenase (3 beta HSD), cytochrome P-450 17 alpha-hydroxylase (P-450c17) and cytochrome P-450 aromatase (P-450arom). A large number of stromal cells were positive for AR, regardless of the distance from a follicle. No steroidogenic enzymes were observed in the stromal cells. In the pre-antral follicle, AR was observed in the theca cells. P-450scc, 3 beta HSD and P-450c17 were sporadically expressed in the theca cells in relatively large-sized pre-antral follicles. ER was positive in the granulosa cells only in the P-450arom-positive antral or pre-ovulatory follicle, which is likely to be a selected follicle. In the corpus luteum, in the period from ovulation to the mid-secretory phase, PR immunoreactivity was observed in a large number of both the luteinized granulosa and the theca cells. All steroidogenic enzymes were observed in all corpora lutea, but ER was negative in any corpus luteum. In the atretic follicle, AR was present in the theca interna cells. P-450scc, 3 beta HSD and P-450c17 were observed in the theca interna cells in some atretic follicles.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxysteroid Dehydrogenases↗