Evaluation of the lymphocyte trafficking drug FTY720 in vaginal tissues.

Background: FTY720 is an immunomodulatory agent that reduces lymphocytes in peripheral tissues and circulation. Such agents may be effective as vaginal microbicides for HIV prevention. Systemic or vaginal application of FTY720 may reduce lymphocyte concentrations in genital tissues, reducing HIV target cell numbers.

Establishment and characterization of a human gestational choriocarcinoma cell line (HOCC).

Human gestational choriocarcinoma cell line (HOCC) was established from the mouse graft of choriocarcinoma. The HOCC cells were spindle or polygonal in shape and multi-nucleated giant cells, showing neoplastic and pleomorphic features. The cell proliferated stably, and the population doubling time was about 32 hours.

Ovarian carcinoid with severe constipation due to peptide YY production.

We report a patient with primary trabecular carcinoid of the ovary with severe constipation probably due to peptide YY production by the tumor. A 43-year-old female had complained of severe constipation for several months and was found to have a left ovarian tumor. The surgically resected tumor was diagnosed as trabecular carcinoid by light microscopic examination.

Localization of fibronectin on the surface of human spermatozoa and relation to the sperm-egg interaction.

Objectives: The mechanism of human sperm-oolemma adhesion and penetration as well as localization of fibronectin on the sperm head and its relation to fertilization were investigated.

Design: Sperm-oolemma interaction was examined with an in vitro assay of the human sperm-zona-free hamster egg interaction. Localization of fibronectin on the surface of human spermatozoa was observed by the back scattered electron imaging mode of the scanning electron microscopy (SEM).

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.

The role of inhibin on pituitary FSH secretion and maturation of pituitary-gonadal axis in immature female rats.

The possible physiological significance of endogenous inhibin was evaluated in prepubertal female rats during sexual maturation. Ovarian/serum inhibin, serum FSH levels were measured from 4th day to 35th day of life by RIA. Serum inhibin levels were first detected on 4th day of life, thereafter, progressively increasing up to 27th day.

Tenascin expression in normal and abnormal human endometrium.

Tenascin has been recently characterized as an extracellular matrix glycoprotein involved in tissue interactions during fetal development and oncogenesis. In order to study the possible involvement of tenascin in epithelial growth of the human endometrium, we evaluated the expression of tenascin in 84 cases of normal, hyperplastic, or neoplastic human edometrium. The specimens were obtained by curettage and/or biopsy and analyzed by immunohistochemistry utilizing a newly developed monoclonal antibody against human tenascin.

Fibronectin in reproduction.

The fibronectin (FN) levels in human follicular fluids have been shown to correlate well with follicular size and oocyte maturity, suggesting a role of FN in oocyte maturation. When added to the culture medium, the synthetic peptide Gly-Arg-Gly-Asp-Ser (GRGDS), which specifically inhibits the cell-binding of FN, has been shown to inhibit both spontaneous resumption of meiosis and gonadotropin-releasing hormone-induced meiosis of the oocytes. In another set of experiments, GRGDS has been found to inhibit the in vitro cleavage of mouse embryos by a still unknown mechanism.

Artificially induced menstrual cycle with natural estradiol and progesterone.

In order to study the feasibility and efficacy of using natural 17 beta-estradiol (E2) and progesterone (P) to induce endometrial changes, a group of patients with history of premature ovarian failure and bilateral oophorectomy, also interested in the embryo transfer program with donor ovum, were given transdermal E2 (Estraderm, Ciba Pharmaceutical Co., Summit, NJ), and vaginal progesterone suppositories. Serial serum E2, P, follicle-stimulating hormone, and luteinizing hormone were determined by radioimmunoassays.

Effects of fibronectin and its peptide fragment on preimplantation mouse embryo.

To investigate the involvement of fibronectin in the early development of mouse embryo, embryos at the two-cell stage were cultured in serum-free Ham F-10 medium with or without fibronectin-related substances. When antibody raised against fibronectin was added at concentrations of 0, 10(-3), 10(-2), and 2 X 10(-2) (vol:vol), the percentages of cells in the blastocyst stage at 72 hours were 88.5%, 67.

Fibronectin and glycosaminoglycans in human preovulatory follicular fluid and their correlation to follicular maturation.

The purpose of this study was to investigate the involvement of two extracellular matrix constituents--fibronectin (FN) and glycosaminoglycans (GAG)--in the development of human ovarian follicles. One-hundred-and-one samples of human follicular fluid (HFF) aspirated from patients participating in an IVF-ET programme were assayed for FN, GAG, protein, progesterone (P) and oestradiol (E2). FN/protein and FN/GAG ratios increased significantly with volume of HFF and with follicular P levels.

Effects of androgen on 17 beta-estradiol production by cultured human granulosa cells.

In order to determine whether androgen acts solely as a substrate for aromatization or whether it also influences on the activity of aromatase enzyme, human granulosa cells were incubated in vitro with or without androgen. Although basal production of 17 beta-estradiol (E2) in cultured granulosa cells obtained from follicles about 26 hr after the initiation of the LH surge was restricted in small quantities, a marked increase in E2 production occurred in the presence of testosterone (T) (10(-6) M) as aromatizable substrate. The non-aromatizable androgen, 5 alpha-dihydrotestosterone (DHT) (10(-7) M, 10(-6) M), slightly enhanced E2 production and it did not inhibit T aromatization in these cells.

[Effects of androgen on 17 beta-estradiol and progesterone secretion by cultured human granulosa cells].

Human granulosa-luteal cells obtained from dominant preovulatory follicles at the time of oocyte retrieval for in vitro fertilization (about 10 h after the peak of the endogenous LH surge) produced large quantities of progesterone (P) (8.7 pg/cell/first 2 days) in a monolayer culture. Although basal production of estradiol (E2) in these granulosa-luteal cells was restricted to small quantities, a marked increase in E2 production occurred in the presence of testosterone (T) (10(-6) M) as aromatizable substrate.

Steroid profiles of follicular fluids from a patient with the empty follicle syndrome.

Steroid profiles of follicular fluid obtained from a patient during a cycle in which no oocytes were recovered in 12 follicular aspirates were compared with those of fluid obtained both from cycles of the same patient and from cycles of other patients when oocytes were recovered. Follicles aspirated in the cycle when no oocytes were recovered were shown to be neither atretic follicles, follicular cysts, nor prematurely luteinized follicles. The steroid profile of follicular fluid from the index cycle was characterized by a markedly increased estradiol-to-progesterone ratio and an increased androstenedione level.

[Ultrastructural study of the intercellular junction and lanthanum penetration in the mouse blastocyst].

Cell junctions in mouse blastocyst were ultrastructurally investigated with or without lanthanum tracer. Tight junctions, gap junctions and desmosomes were observed in the trophectoderm. The tight junction was located near the zona pellucida in all trophoblast interspaces, whereas the gap junction and the desmosome, which were infrequently observed, were localized far from the zona pellucida.

Sperm-egg interactions observed by scanning electron microscopy.

Successful in vitro fertilization requires mature oocytes in which the first polar body has been extruded and capacitated sperm capable of penetrating the zona pellucida. In this study we made a time sequential observations on human sperm-egg interactions by SEM in two experimental systems. Human sperm-human zona pellucida interaction: Cytoplasmic processes of corona cell extend around sperm head.

[The evaluation of serum estradiol, progesterone, testosterone concentrations and urinary estrogen excretion level for the monitoring indices of follicular maturation].

We have evaluated serum estradiol, progesterone, testosterone, and urinary estrogen excretion in 24 hour urine samples to monitor indices of follicular maturation. The serum steroid levels were determined with the direct radioimmunoassay kit. The urinary estrogen level was measured with the estrogen micrometering kit using hemagglutination inhibition reaction.

Relationship between follicular fluid steroid concentrations and in vitro fertilization.

Steroid concentrations were measured in follicular fluid obtained from patients fertilized in vitro. Progesterone and estradiol-17 beta concentrations showed positive correlations with the growth of follicles, whereas the testosterone concentration had negative correlations not only with follicle growth but also with progesterone and estradiol-17 beta concentrations. The testosterone concentration was significantly lower in follicles with cleaved oocytes than in those with uncleaved oocytes or those with cleaved but degenerated oocytes.

[The influence of various molecules on the occurrence of the human sperm acrosome reaction].

The acrosome reaction is essential for fertilization, but the mechanism of the acrosome reaction of human spermatozoa is not clear at the present time. We studied the mechanism to analyze the cause of unexplained infertility, the appropriate timing of insemination, and the environment of spermatozoa prior to fertilization. For this study, we examined the effects of Ca++, Mg++, Kallikrein, Phospholipase A2, p-bromophenacyl bromide (Phospholipase A2 specific inhibitor), Lysophosphatidyl choline, Arachidonic acid, and Glyceryl monooleate using in vitro penetration assay employing zona- free hamster eggs.

[Effects of the addition of serum albumin on penetration of human spermatozoa into zona-free hamster eggs].

Since Yanagimachi et al. (1976) suggested that human spermatozoa were capable of penetrating into zona-free hamster eggs, this in vitro assay has been used to analyse the fertilizing ability of human spermatozoa. Serum albumin is an important constituent of the medium used for the assay.

Effects of phospholipase A2, lysophosphatidyl choline, and fatty acid on the acrosome reaction of human spermatozoa.

An in vitro penetration assay employing zona-free hamster eggs was used to study the effects of phospholipase A2, lysophosphatidyl choline, and fatty acid on the acrosome reaction of human spermatozoa. Human spermatozoa were preincubated for 4 hr in modified Biggers, Whitten, and Whittingham's medium (mBWW) containing a specific phospholipase A2 inhibitor, p-bromophenacyl bromide (p-BPB: 1 X 10(-5) - 1 X 10(-3) M), lysophosphatidyl choline (LC: 5-500 micrograms/ml), and arachidonic acid (AA: 5-500 micrograms/ml), prior to the addition of zona-free superovulated hamster eggs. Eggs were examined microscopically 2 or 4 hr later for evidence of swelling or decondensing sperm heads in the cytoplasm.

Induction of ovulation with an estrogen antagonist, tamoxifen.

Tamoxifen (Norvadex), synthesized by the Imperial Chemical Industries Ltd., is a triphenylethylene derivative having a clomiphene-like structure and displays anti-estrogenic activities. In this study we used tamoxifen for the treatment of anovulatory infertility and investigated the clinical results.

Measurement of urinary estrogen level with the urinary estrogen micrometry method in in vitro fertilization and embryo transfer therapy.

We have evaluated a new micrometering kit using hemagglutination inhibition reaction for determination of total estrogens in urine of normal fertile women and women undergoing in vitro fertilization and embryo transfer (IVF-ET). The micrometering kit was sensitive enough to monitor follicular maturation, and its technique was simple and rapid. Results were obtained in 2.

[Prediction of Ovulation and timing of AIH].

The relation among ultrasonographic follicular diameter, serum estradiol (E2), luteinizing hormone (LH), cervical mucus changes, and basal body temperature (BBT) was studied. One hundred and eight cycles were examined in forty-two infertile patients. The last low day of the BBT curve was taken as day zero.