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Biomedical subjects

T Fukaya

Publications and source records attributed to T Fukaya.

At least 19 recordsLinked to original sources
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

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

[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↗
PubMed1994

Substance P induces inositol 1,4,5-trisphosphate and intracellular free calcium increase in cultured normal human epidermal keratinocytes.

Substance P is a neuropeptide which is present in peripheral C nerve endings and released from them. Free nerve endings of C nerve are present in human epidermis. The effects of substance P on the transmembrane signaling system of pig epidermal sheets were previously reported. In these studies, a small amount of cells other than keratinocytes contaminated the epidermal sheets and the species difference from human was also noticed. Therefore we investigated the effects of substance P on cultured normal human epidermal keratinocytes. Alteration of intracellular free calcium (Ca2+) in single living keratinocytes was studied using an inverted fluorescence microscope and Ca(2+)-sensitive dye, Fura 2-AM. Treatment of normal human epidermal keratinocytes with substance P resulted in an increase in inositol 1,4,5-trisphosphate and in intracellular Ca2+. Substance P inhibited DNA synthesis of the keratinocytes in a dose-dependent manner. These results are consistent with the view that substance P stimulates phosphatidylinositol-4,5-bisphosphate hydrolysis of human keratinocytes, resulting in inositol 1,4,5-trisphosphate-Ca2+ signal.

Calcium↗
PubMed1994

Quantitation of P450 aromatase immunoreactivity in human ovary during the menstrual cycle: relationship between the enzyme activity and immunointensity.

To understand changes associated with the menstrual cycle in the human ovary, it is very important to examine chronological changes in P450 aromatase (P450arom) enzymatic activity in the normal cycling ovary. Therefore, we initially examined the correlation between intensity of P450arom immunoreactivity and its biochemical enzymatic activity in five estrogen-producing human cancer cell lines (HHUA, Ishikawa, HEC-59, OMC-2, and MCF-7). P450arom immunointensity per cell was evaluated by the CAS 200 computed image analysis system, and its catalytic activity per 10(6) culture cells was analyzed by the tritiated water method. A significant correlation (r = 0.959) was demonstrated between P450arom immunoreactivity and enzymatic activity under optimal conditions of tissue fixation and immunohistochemical procedures. We then investigated P450arom immunointensity in 31 specimens of normal cycling human ovaries to examine chronological changes in P450arom activity per cell throughout the menstrual cycle. In the follicular phase, P450arom was observed in the granulosa cells of one selected antral follicle per case during the mid- to late proliferative period, and its immunointensity per granulosa cell in the follicle was not significantly different between mid- and late proliferative periods, although serum estradiol level was markedly elevated in the late proliferative period. In the luteal phase, both P450arom immunointensity per luteinized granulosa cell in a corpus luteum and serum estradiol level reached a peak in the mid-secretory period. These findings indicate that different factors may influence ovarian P450arom activity during the follicular and luteal phases, i.e., an increased number of granulosa cells in the selected follicle during the follicular phase but changes in P450arom activity per luteinized granulosa cell in the corpus luteum during the luteal phase.

3,3'-Diaminobenzidine↗
PubMed1994

Influence of HLA antigens on reproduction among Japanese population: study of haplotypes in 247 families.

Although an increased incidence of shared HLA antigens among couples experiencing recurrent spontaneous abortion or infertility has been observed, antigenic effects remain to be clarified. These effects, which are suspected to exist among fertile couples, were examined in this study. A group of 247 healthy Japanese families and their 682 children, whose HLA antigen haplotypes could be determined, were studied. Haplotypes of the fathers were compared with those of the mothers, and the combination of HLA antigens in the A, B, C and DR loci was determined as antigenic patterns against the mother. Patterns inherited by children were examined and deviations of the patterns were studied according to the natural rule that either of the haplotypes should be inherited at equal rates. Lower rates of children with antigens common with those of the mother, which may show a result of reproductive selection, were observed among only the first born children in the A (p < 0.05) and DR loci (p < 0.01), but rates similar to those expected were observed among the second born and later children. Lower rates were also observed in the B and C loci among all children with no significant difference between groups of the children. The necessity of differing antigenic factors in order to achieve pregnancy in the population was shown. Important effects of HLA antigenicity in reproductive failure are suggested.

Adult↗
PubMed1994

Is treatment of long-term and consecutive use of clomiphene citrate effective in anovulatory patients? Results of multi-centric retrospective studies.

The efficacy of long term consecutive treatment of clomiphene citrate (CC) was investigated in 608 infertile women associated with anovulatory disorders (classified as WHO group II amenorrhea). This study was performed in multi-centric facilities of obstetrics and gynecology by a retrospective randomized assessment. Infertile women associated with anovulation were treated by 50-150 mg of CC for five consecutive days in each treatment cycle. In this study, 200 pregnancies were observed and the total pregnancy rate was 32.9%. The number of abortions were 33 out of the 200 pregnancies (16.5%). The cumulative pregnancy rate within the pregnant subjects reached 90% in initial 10 treatment cycles and there was no difference between 50 mg and 100 mg of CC treatment. There were remarkable differences among the pregnancy rates achieved during consecutive treatments at the primary (clinics and hospital), secondary (regional center hospitals) and advanced facilities. Consecutive treatment by CC for infertile women associated with anovulation may be effective during initial 10 cycles. Although ovulation can be confirmed during the following cycles, the stimulation protocol should be reconsidered after 12 consecutive cycles therapy with CC, because cycle fecundity decreases. Hence, to improve pregnancy rates, it is important to check the previous administration of CC which reflects adversely on the subsequent conception rate especially in advanced facilities where these patients are referred to.

Anovulation↗
PubMed1994

The role of macrophage colony stimulating factor in the peritoneal fluid in infertile patients with endometriosis.

Macrophage colony stimulating factor (M-CSF) concentrations in the peritoneal fluid of 44 infertile patients were measured. The concentration of M-CSF was significantly higher in the patients with endometriosis than in those without endometriosis. There was no significant difference between the M-CSF concentration and stage of endometriosis classified by revised American Fertility Society (rAFS) classification in the 44 patients. However, in primary infertility patients with endometriosis there was a significant correlation between peritoneal M-CSF levels and rAFS points of endometriosis. We speculate that there is some relationship between the increased levels of M-CSF in the peritoneal cavity and the primary infertility associated with endometriosis especially progression of endometriosis.

Ascitic Fluid↗
PubMed1993

Laparoscopic evaluation of the onset and progression of endometriosis.

OBJECTIVE: To clarify the pathogenesis of endometriosis on the basis of analysis of primary lesion sites, age at onset, rate of progression, and response to drug treatment. STUDY DESIGN: The clinical records of 690 women with laparoscopically confirmed endometriosis were retrospectively analyzed based on the revised American Fertility Society point system. RESULTS: The primary site of endometriosis was the uterosacral ligament and pelvic peritoneum/pouch of Douglas in 73% of patients with stage I disease, whereas only 16% had ovarian lesions. However, disease progression was associated with an increasing frequency of ovarian lesions. In terms of the revised American Fertility Society score, endometriosis progressed at a mean rate of 0.3 point per month. Thus the earliest onset of endometriosis was estimated at 3 to 4 years after menarche. Drug therapy improved the revised American Fertility Society score by about 50%. Patients with a low response to an initial cycle of therapy generally showed further improvement after an additional treatment cycle. CONCLUSIONS: Because endometriosis may occur as early as 3 to 4 years after menarche and gradually progresses, drug therapy, including long-term treatment, should be carried out in women with definitive evidence of endometriosis who must maintain their reproductive potential.

Adolescent↗
PubMed1993

Is pelvic endometriosis always associated with chronic pain? A retrospective study of 618 cases diagnosed by laparoscopy.

OBJECTIVE: To establish a relationship between pelvic pain and the stage and severity of pelvic endometriosis determined according to the revised American Fertility Society classification. STUDY DESIGN: Retrospective study of 618 cases diagnosed by laparoscopy as endometriosis at the Tohoku University Hospital from 1984 to 1991. RESULTS: Pelvic pain did not appropriately reflect the stage and severity of pelvic endometriosis in 40% of patients. Although the pain in some patients in stage IV showed some correlation with the stage and severity of the disease, others had either no symptoms or paradoxically complained of less pain than those patients with stage I or II endometriosis. CONCLUSIONS: With the present revised American Fertility Society classification of endometriosis we cannot evaluate the severity of pelvic pain.

Chronic Disease↗