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Biomedical subjects

H Suginami

Publications and source records attributed to H Suginami.

At least 19 recordsLinked to original sources

Familial risk among Japanese patients with endometriosis.

OBJECTIVES: This study was designed to examine the prevalence of endometriosis among female siblings of patients with endometriosis in Japan. METHODS: A total of 339 patients with endometriosis were questioned about endometriosis in their sisters. The control group consisted of 284 Japanese healthy fertile women with no history of endometriosis. Similarly, the controls were interviewed about their sisters. RESULTS: We detected sisters with endometriosis in 8.8% of cases and 1.5% of the control population. The relative risk of endometriosis in female siblings was 5.7. However, a significant difference was not seen in age at diagnosis and clinical stage between patients with or without a family history of endometriosis. CONCLUSIONS: These data demonstrate a familial tendency for endometriosis and suggest that endometriosis has a genetic factor in the pathogenesis.

Adolescent↗

Laparoscopic surgery for endometriosis: a long-term follow-up.

OBJECTIVE: To investigate if complete resolution of endometriosis by laparoscopic surgery is beneficial to postoperative fecundity, dysmenorrhea and dyspareunia. DESIGN: An observational comparative study on the outcome of laparoscopic surgery. PATIENTS: Laparoscopically-treated symptomatic women with endometriosis (total n = 236); complete (n = 185) and incomplete (n = 51) surgery groups. MEASUREMENTS: Postoperative fecundity and symptom reduction. RESULTS: With whole populations, no surgical completeness-related difference was observed in cumulative pregnancy rates during the postoperative days 0-400 (cycle fecundity rate = 0.0319). Further accumulation of pregnant cases was followed in the complete surgery group (final cumulative pregnancy rate = 80%), but not in the counterpart group (p = 0.003). The similar result was obtained when only r-AFS classification stages III and IV were compared (p = 0.007). No r-AFS stage-related difference was observed in cumulative pregnancy rates when only patients of complete surgery were selected for comparison. The surgery reduced dysmenorrhea (84.7%) and dyspareunia (80.0%). CONCLUSIONS: Laparoscopic conservative surgery for endometriosis, especially when it is complete, increases fecundity and reduces disease-related symptoms, such as dysmenorrhea and dyspareunia.

Adult↗

Cell adhesion and reproduction. An overview.

Cell adhesion is essential for the regulation of many cellular functions. The adhesion molecule plays a critical role as a fundamental substance in various processes of reproduction such as trophoblast-endometrial interaction. Our understanding of the physiology of adhesion molecules will be helpful for clinical application of these molecules in the treatment and assessment of disorders of many processes of reproduction. We will briefly review integrins, cadherins, and laminin, which were the principal subjects of discussion in this conference.

Cadherins↗

Physiological roles of integrin alpha 6 beta 1 in ovarian functions.

We previously reported that human ovarian cells express integrin beta 1 families. The physiological role(s) of integrins was investigated using in vitro human granulosa cell culture and in vivo mouse ovulation model. In human luteinizing granulosa cell culture obtained from the patients undergoing in vitro fertilization treatment, laminin, which is a ligand for integrin alpha 6 beta 1, suppressed the production of progesterone by granulosa cells. On the other hand, the anti-alpha 6 monoclonal antibody (mAb) GoH3, which partially inhibits the interaction between integrin alpha 6 beta 1 and laminin, enhanced production of progesterone by 2-fold of the control under the culture with laminin, indicating that integrin alpha 6 beta 1 regulates the luteinization of human granulosa cell during the periovulatory phase. In an immature superovulated 13-day-old ICR (CD-1) mice model, intraperitoneal administration of GoH3 induced successful ovulation, whereas no ovulation was observed in the GoH3-nontreated groups, showing that integrin alpha 6 beta 1 is related to gonadotropin-induced follicular growth. These findings suggest that the interaction between integrin alpha 6 beta 1 and laminin plays an important role in the corpus luteum formation and follicular growth.

Animals↗

Bestatin, an aminopeptidase inhibitor, promotes follicular growth and ovulation suppressed by stress in mice.

We previously reported that the intraperitoneal and intrabursal administration of bestatin, an aminopeptidase inhibitor, enhanced follicular growth and ovulation in immature mice. In this study, we examined the effect of bestatin on follicular growth and ovulation under the conditions in which the ovarian response to gonadotropins was suppressed. Immature female mice were exposed to 72-h continuous lighting (on days 20-22 of life), or to 24-h starvation (on day 20). On days 20-22, bestatin (4 mg/ml, 100 microl) or PBS (100 microl) was administered ip. 4 times and pregnant mare serum gonadotropin (PMSG, 5 IU) and hCG (5 IU) were given on days 21 and 23, respectively. The numbers of ovulated oocytes per mice were significantly attenuated by both stresses. Bestatin significantly enhanced the numbers of ovulated oocytes under both lighting (49.5+/-7.0 vs. 28.0+/-5.5) and starvation (28.6+/-2.6 vs. 20.2+/-2.0) stresses (with vs. without bestatin, P<0.05). When follicular development was stimulated by PMSG (20 IU) on day 21, the serum estradiol concentration on day 23 was also suppressed by both stresses, but enhanced by bestatin under lighting (231.3+/-36.0 vs. 111.0+/-23.0 pg/ml) and starvation (151.9+/-8.8 vs. 90.7+/-15.4 pg/ml) stresses (with vs. without bestatin, P<0.01). Northern blot analysis revealed that the expression of P450arom-mRNA in the ovaries induced by PMSG (20 IU) was increased on day 23 by treatment with bestatin under starvation stress. These results indicated that stress inhibited the ovarian response to gonadotropins and that bestatin restored the response suppressed by stress.

Aminopeptidases↗

An aminopeptidase inhibitor, bestatin, enhances progesterone and oestradiol secretion by porcine granulosa cells stimulated with follicle stimulating hormone in vitro.

We examined the presence of cell surface aminopeptidase on cultured porcine granulosa cells by employing the aminopeptidase assay using alanine-p-nitroanilide and histochemical staining using L-leucyl-beta-naphthylamide. Porcine granulosa cells obtained from follicles 4-5 mm in diameter were cultured for 7 days. The aminopeptidase assay showed that the porcine granulosa cell culture had aminopeptidase activity and that this activity was inhibited in a dose-dependent manner by bestatin which binds to cell surfaces and inhibits cell surface aminopeptidases. Histochemical staining also indicated that cultured granulosa cells had aminopeptidase activity. Porcine granulosa cells were cultured in the presence or absence of porcine follicle stimulating hormone (FSH, 3.125 nmol/l) and/or bestatin (0.4, 4.0 and 40.0 micrograms/ml) for 7 days, and the production of progesterone and oestradiol was measured. In the presence of porcine FSH, the production of progesterone and oestradiol by granulosa cells was increased significantly by approximately 5- and 2-fold respectively. These increases were enhanced further by bestatin (40.0 micrograms/ml). In the absence of porcine FSH, progesterone production was enhanced by bestatin (40.0 micrograms/ml), whereas no significant effect of bestatin on oestradiol secretion was observed. These findings indicate that the inhibition of membrane-bound aminopeptidase(s) on the cell surfaces affects the steroidogenesis of granulosa cells, and that these aminopeptidase(s) are important regulators of granulosa cell differentiation.

Aminopeptidases↗

An aminopeptidase inhibitor, bestatin, enhances gonadotrophin-stimulated ovulation in mice.

The role of aminopeptidases in follicular growth and/or ovulation in vivo was examined. We injected an inhibitor of cell-surface aminopeptidases, bestatin (4 mg/ml, 100 microliters), i.p. four times during 2 days into 20 day old female ICR mice, in which follicular growth and ovulation were stimulated by pregnant mare's serum gonadotrophin (PMSG, 5 IU) and human chorionic gonadotrophin (HCG, 5 IU). The number of ovulated oocytes was estimated by counting the number of oocytes in the oviduct 19 h after HCG injection. The mean +/- SD number of ovulated oocytes in bestatin-treated mice was significantly higher than that in control mice [47.00 +/- 18.13 (n = 26) versus 35.90 +/- 10.14 (n = 28), P < 0.01]. To confirm the direct effect of bestatin on the ovary, bestatin (2 mg/ml, 3 microliters) or its stereoisomer (2 mg/ml, 3 microliters) with very weak inhibitory activity was unilaterally injected into the ovarian bursa 24 h before the administration of PMSG. As a control, buffer (3 microliters) was injected into the contralateral bursa. In some experiments, bestatin (2 mg/ml, 3 microliters) was injected just before HCG administration. The administration of bestatin via the ovarian bursa prior to PMSG administration significantly increased the number of ovulated oocytes per oviduct from the treated compared with the contralateral ovary [23.70 +/- 9.61 versus 17.10 +/- 5.83 (n = 25), P < 0.01], whereas its stereoisomer elicited no significant effects. The administration of bestatin just before HCG administration also had no effect. These findings indicate that membrane-bound peptidase(s) present on murine ovarian cells is an important regulating factor(s) of follicular growth and/or ovulation.

Aminopeptidases↗

Granulosa cells express integrin alpha 6: possible involvement of integrin alpha 6 in folliculogenesis.

To identify differentiation antigens of granulosa cells, we have raised two monoclonal antibodies against porcine granulosa cells (POG-2 antibody) and human granulosa cells (OG-1 antibody). Analysis of N-terminal amino acid sequences of purified OG-1 and POG-2 antigens revealed that they were identical to human integrin alpha 6 and its porcine homolog, respectively. An immunohistochemical study showed that integrin alpha 6 was highly expressed on granulosa cells in preovulatory follicles in the human ovary, whereas it was highly expressed on granulosa cells in small follicles in the porcine ovary. As a common characteristic of integrin alpha 6 expression in both ovaries, granulosa cells located in inner layers, which are not in contact with the basal lamina, expressed integrin alpha 6 on the cell surface. This expression profile suggests that the interaction between integrin and extracellular matrices occurs in the granulosa cells located in inner layers. This cell surface interaction is reported to modulate soluble signals to the cells. Since the expression of integrin alpha 6 on inner-layer granulosa cells is related to stages of differentiation, this interaction may be involved in folliculogenesis.

Animals↗

Micro-bead embolization of uterine spiral arteries and changes in uterine arterial flow velocity waveforms in the pregnant ewe.

The aim of the present study was to simulate the pathological uteroplacental circulation observed in complicated human pregnancies in the pregnant ewe and to analyze its velocimetric changes. Four pregnant ewes at 16-17 weeks of pregnancy were used in the study. Micro-beads (Gelfoam) were administered stepwise into the uterine artery and changes in the uterine circulation were assessed by Doppler velocimetry. Gelfoam administration successfully embolized the uterine spiral arteries located in the decidual segment. The Gelfoam embolization decreased the uterine blood flow dose-dependently from 550 +/- 48 ml/min (mean +/- SD) to 142 +/- 12 ml/min and reciprocally increased the uterine vascular resistance from 139 +/- 12 mmHg min l-1 to 540 +/- 46 mmHg min l-1 at 0 mg and 30 mg Gelfoam, respectively. It dose-dependently attenuated the pregnancy-related physiological elevation in diastolic flow velocity, while the systolic flow velocity was unaffected, resulting ina dose-dependent increase in the pulsatility index from 0.5 +/- 0.2 to 3.2 +/- 0.7 at 0 mg and 30 mg Gelfoam, respectively. The pulsatility index linearly correlated with the uterine vascular resistance, giving a high correlation coefficient of r = 0.947. It could be concluded that the uterine arterial pulsatility index is an indicator of uterine vascular resistance.

Animals↗

The effect of follicular fluid on intracellular free calcium levels in human spermatozoa.

In this study, we demonstrate the effect of follicular fluid (FF) on free cytosolic calcium ion ([Ca2+]i) in human spermatozoa, using a fluorescent indicator "fura 2," and we evaluate the factors that increase [Ca2+]i in FF. A significant and immediate [Ca2+]i increase (656.3 +/- 148.6 nM) was elicited by intact FF. The increase in [Ca2+]i elicited by lipid-stripped FF was only 50% (322 +/- 68.7 nM) of that elicited by intact FF. However, 82% of the [Ca2+]i increase elicited by intact FF was observed when lipid-stripped FF was supplemented with progesterone, but not when it was supplemented with estradiol. Also, progesterone at the concentrations of 1 ng/ml to 1 microgram/ml caused a significant increase in [Ca2+]i by itself, but estradiol produced a small effect, ranging from 3% to 9% of the effect produced by progesterone. These results indicate that progesterone plays a major role in the lipid fraction in FF to elicit the entry of calcium into human spermatozoa.

Calcium↗

A case of laparoscopic complication: injury of the left common iliac vessels and subsequent acute compartment syndrome of the left leg.

A 45-year-old Japanese woman underwent a laparoscopy-assisted vaginal hysterectomy. Insertion of a trocar injured the left common iliac artery and vein, which were repaired within 2.5 hours. Postoperatively the patient presented an acute compartment syndrome of the left leg. Fasciotomy and rehabilitation rescued her from functional disturbances.

Acute Disease↗

Expression and localization of aminopeptidase N, neutral endopeptidase, and dipeptidyl peptidase IV in the human placenta and fetal membranes.

OBJECTIVE: Our purpose was to determine the distribution of membrane-bound cell surface peptidases, namely aminopeptidase N, neutral endopeptidase, and dipeptidyl peptidase IV in the human placenta and fetal membranes. STUDY DESIGN: Frozen tissue sections of the first-trimester chorionic villi, term placentas, and term fetal membranes were stained by indirect immunofluorescence with specific monoclonal antibodies. RESULTS: In the first trimester chorionic villi cytotrophoblasts expressed both neutral endopeptidase and dipeptidyl peptidase IV, but syncytiotrophoblasts expressed only neutral endopeptidase. Stromal cells in the chorionic villi expressed the three peptidases at various intensities. In the term placentas villous syncytiotrophoblasts expressed neutral endopeptidase weakly, and the villous stromal cells expressed large amounts of both aminopeptidase N and dipeptidyl peptidase IV but neutral endopeptidase weakly or faintly. In the term fetal membranes amniotic epithelial cells and chorion laeve expressed both neutral endopeptidase and dipeptidyl peptidase IV. Decidual cells in the decidua parietalis moderately or highly expressed aminopeptidase N. CONCLUSION: Three peptidases, aminopeptidase N, neutral endopeptidase, and dipeptidyl peptidase IV, are expressed by different cell populations in the human placenta and fetal membranes, suggesting their respective and important roles at the maternofetal interface.

Aminopeptidases↗

A new monoclonal antibody (POG-1) detects a differentiation antigen of porcine granulosa and thecal cells and indicates heterogeneity of thecal-stromal cells.

To identify the differentiation antigen of ovarian cells, we raised a murine monoclonal antibody (POG-1 antibody) reactive to porcine granulosa and thecal cells in the ovary. Immunofluorescence staining showed that expression of the POG-1 antigen on granulosa and theca interna cells increased gradually in accordance with follicular development. The thecal cells just outside the basal lamina surrounding the follicles did not express the antigen, whereas some stromal cells around the theca externa layer in the large follicles did express it. These expression profiles indicated the heterogeneity of thecal-stromal cells and that the POG-1 antigen was a differentiation-related antigen of granulosa and thecal cells. Luteal cells also expressed the antigen. In organs other than the ovary, some endocrine and exocrine cells, such as Leydig cells and secretory cells of the breast, expressed the antigen. The POG-1 antigen was purified from granulosa cells by immunoaffinity chromatography. Polyacrylamide gel electrophoresis profiles showed that the antigen consisted of two specific proteins; the major one had a molecular mass of 77, and the other had a molecular mass of 81 kilodaltons. Analysis of the purified POG-1 antigen may contribute to understanding the differentiation mechanism of granulosa and thecal cells.

Animals↗

Cytokines stimulate dipeptidyl peptidase-IV expression on human luteinizing granulosa cells.

We have previously reported that dipeptidyl peptidase-IV (DPPIV) is a differentiation antigen for human granulosa cells that is initially expressed during corpus luteum formation. To investigate the involvement of cytokines in luteal cell differentiation, we examined the expression and activity of DPPIV in human luteinizing granulosa cells cultured in vitro. Human granulosa cells obtained from patients who had undergone in vitro fertilization were cultured for 7 days in the absence (controls) or presence of hCG (1 U/mL), tumor necrosis factor-alpha (TNF alpha; 10 ng/mL), or interleukin 1-alpha (IL-1 alpha; 10 ng/mL). Flow cytometry showed that the percentage of cultured granulosa cells treated with TNF alpha and IL-1 alpha that was positive for DPPIV expression was significantly higher than that in controls (43.7 +/- 5.4% and 43.4 +/- 5.6%, respectively, vs. 21.7 +/- 3.5%; P < 0.01), whereas hCG treatment produced no remarkable difference in DPPIV expression (24.0 +/- 5.2%). The DPPIV activity of cells treated with TNF alpha and IL-1 alpha was also significantly higher than that of controls, whereas hCG treatment produced no significant difference from control values. These findings indicate that TNF alpha and IL-1 alpha stimulate DPPIV expression and activity in human luteinizing granulosa cells in vitro and suggest the involvement of cytokines in the differentiation of granulosa cells during corpus luteum formation.

Adult↗

A clomiphene citrate and tamoxifen citrate combination therapy: a novel therapy for ovulation induction.

OBJECTIVE: To assess the efficacy of a clomiphene citrate (CC) and tamoxifen citrate (CC/tamoxifen) combination therapy in ovulation induction by comparing with a CC alone therapy. DESIGN: A randomized cross-over study with CC alone and CC/tamoxifen combination therapies for 20 normoprolactinemic anovulatory women. Randomly selected 10 of the 20 women (group A) underwent a CC alone therapy (100 mg CC during cycle days 5 to 9) for the initial three consecutive treatment cycles and a CC/tamoxifen combination therapy (50 mg CC and 20 mg tamoxifen during cycle days 5 to 9) for the subsequent three consecutive treatment cycles. The remaining 10 (group B) were treated similarly but with the inverse sequence. Ovulation was documented when a high BBT phase persisted for 11 days or more, when midluteal serum P levels exceeded 7 ng/mL (22.3 nmol/L), or when pregnancy ensued. RESULTS: The rates of ovulatory/treated, pregnant/treated, and pregnant/ovulatory cycles were 42 of 56 (75.0%), 3 of 47 (6.4%), and 3 of 35 (8.6%) for CC/tamoxifen, respectively. Those were 25 of 57 (43.9%), 1 of 48 (2.1%), and 1 of 21 (4.8%) for CC, respectively. Clomiphene citrate/tamoxifen was more effective in ovulation induction than CC. All the pregnancies were normal and single. None of the treatments was accompanied by any remarkable side effects. CONCLUSION: Clomiphene citrate/tamoxifen is a novel treatment modality of ovulation induction with high efficacy.

Adult↗