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Biomedical subjects

S F Gould

Publications and source records attributed to S F Gould.

At least 19 recordsLinked to original sources
PubMed2000

Corticotropin-releasing hormone increases the expression of the prostaglandin E(2) receptor subtype EP1 in amnion WISH cells.

The purpose of this study was to investigate the effect of corticotropin-releasing hormone (CRH) on the expression of the prostaglandin (PG) E(2) EP1 receptor subtype and PGE(2) production in amnion WISH cells (AWC). AWC cultures were incubated with CRH. Culture fluid was collected for PGE(2) measurement, and the cells were collected and analyzed for EP1 protein and mRNA. Immunohistochemical localization of the EP1 receptor was also performed. Incubation of AWC with CRH resulted in a dose-dependent increase (r = 0.97) in the level of EP1 receptor protein (P < 0.001). Coincubation of AWC with CRH and indomethacin resulted in the decreased production of PGE(2) while having no effect on EP1 receptor expression. A significant but not dose-dependent increase in EP1 mRNA expression was also observed (P < 0.01). Immunohistochemical evaluation verified cell membrane localization of the receptor in both stimulated and unstimulated cells and confirmed the increased expression of EP1 receptor in response to CRH. Incubation of AWC with CRH also resulted in increased culture fluid PGE(2) levels (P < 0.01). These results suggest that the role CRH plays in the initiation of labor may also involve the promotion of elevated PGE(2) levels and increased expression of the EP1 receptor in amnion.

Amnion↗
PubMed1999

Modulation of the prostaglandin E receptor: a possible mechanism for infection-induced preterm labor.

OBJECTIVE: To evaluate the modulatory effects of interleukin (IL)-1beta and prostaglandin (PG)E2 on the PGE2 receptor subtype EP1 in amnion cell cultures. METHODS: Amnion cell cultures were incubated in increasing concentrations of (IL)-1beta or PGE2. Cultures were also incubated in high concentrations of IL-1beta and PGE2 in combination. Changes in EP1 receptor levels were evaluated by western and northern blot analysis. Culture fluid PGE2 levels were measured by enzyme-linked immunosorbent assay. RESULTS: EP1 receptor protein levels decreased with increasing levels of PGE2 (r = -0.82, P < .05). EP1 receptor protein (r = 0.95, P < .05), EP1 mRNA (r = 0.95, P < .01), and culture fluid PGE2 levels (P < .01) were all increased after IL-1beta administration. EP1 receptor levels also increased approximately fourfold in response to IL-1beta incubation even in the presence of high agonist (PGE2) concentrations (P < .01). CONCLUSION: The results of this study show that IL-1beta might be involved in infection-induced preterm labor by interfering with the normal regulation of EP1 receptor levels and with the promotion of increased PGE2 production in amnion tissue.

Cells, Cultured↗
PubMed1999

Immunohistochemical localization of the prostaglandin E subtype-1 receptor in cytokine-stimulated and unstimulated amnion cells.

OBJECTIVE: To visualize histochemically the prostaglandin EP1 receptor in human amnion cells and to study the effect of inflammatory cytokines, which are known to stimulate the EP1 receptor, on localization. METHODS: Immortalized amnion cells, grown on standard microscope slides and either nonstimulated (control) or stimulated by incubation in culture medium containing interleukin-1beta (25 ng/mL), interleukin-4 (50 ng/mL), or tumor necrosis factor alpha (25 ng/mL), were incubated with rabbit anti-human EP1 antibody and stained by a two-step indirect immunoperoxidase strepavidin-biotin method using horseradish peroxidase and 3,3' diaminobenzidine as the chromogen. The localization was done on ten different flasks of cells. Duplicate slides for each cytokine concentration were prepared. Negative controls for each reagent, prior blocking with 1% bovine serum albumin or 1% milk, or pretreatment with preimmune rabbit immunoglobulin G were run simultaneously. Slides were viewed by standard light microscopy with and without counterstaining with hematoxylin. RESULTS: Amnion cells incubated in medium alone showed receptor localization throughout the cytoplasmic region of the cell membrane. The localization was nonuniform; a discrete unipolar region of perinuclear nonlocalization was observed. Staining occurred in widely dispersed nests. Cytokine stimulation resulted in increased intensity of staining and an increase in the size of the positive nests; however, it did not affect the discrete unipolar perinuclear region of nonlocalization. CONCLUSION: Histochemical localization of the human EP1 receptor confirms a cytoplasmic identity and probable plasma membrane localization. Stimulation by inflammatory cytokines increases staining by recruitment of new amnion cells and appears to increase receptor density per cell.

Amnion↗
PubMed1999

Antepartum and postpartum Papanicolaou smears. Are they both necessary?

OBJECTIVE: To determine if a Papanicolaou smear performed during pregnancy needs to be repeated at the postpartum visit. STUDY DESIGN: A retrospective chart review was performed on women who received prenatal care from our obstetric clinic and returned for a postpartum visit. All pregnant women who had both a Papanicolaou smear in pregnancy and another at the postpartum visit, about six weeks after delivery, were included. Six hundred one patients were studied who had complete records between 1995 and 1996. RESULTS: The average length of time between the two smears was 209 days. With the exception of a lower frequency of the presence of endocervical cells in the antepartum Papanicolaou smears, there were no differences between the distribution of Papanicolaou smear results. In patients without risk factors and a normal antepartum Papanicolaou smear, the prevalence of squamous intraepithelial lesion on the postpartum smear was very low (4 in 1,000). CONCLUSION: In patients without risk factors for cervical intraepithelial neoplasia and a normal Papanicolaou smear in pregnancy, a Papanicolaou smear should be repeated at the recommended yearly interval.

Adult↗
PubMed1998

Tumor necrosis factor-alpha upregulates the prostaglandin E2 EP1 receptor subtype and the cyclooxygenase-2 isoform in cultured amnion WISH cells.

Recent studies have demonstrated a strong correlation between infection and preterm labor. Preterm delivery is also associated with high levels of cytokines and prostaglandins in amniotic fluid. The purpose of this study was to investigate the effect of tumor necrosis factor-alpha (TNF-alpha) on the levels of cyclooxygenase, prostaglandin E2 production (PGE2), and expression of the PGE2 receptor subtype EP1 in amnion WISH cell culture. Amnion WISH cell cultures were incubated in increasing concentrations of TNF-alpha (0-50 ng/ml). Changes in cyclooxygenase and EP1 receptor proteins were evaluated by Western blot analysis. Changes in EP1 mRNA were evaluated by Northern blot, and culture fluid concentrations of PGE2 were estimated by enzyme immunoassay (EIA). EP1 protein (p<0.01), EP1 mRNA (p<0.05), cyclooxygenase-2 (COX-2) protein (p<0.001), and PGE2 concentrations (p<0.01) all increased with increasing concentrations of TNF-alpha. Changes in COX-1 protein were not observed following TNF-alpha-incubation. The results suggest that TNF-alpha may play a role in infection-induced preterm labor by its pleiotropic ability to simultaneously stimulate COX-2 activity, PGE2 concentrations, and PGE2 EP1 receptor levels in human amnion.

Amnion↗
PubMed1985

Effect of estrogen on the affinity of malachite green for staining cardiac lipid inclusions in mice.

The effect of estradiol-17-beta on lipids of the ventricular myocardium of mice has been studied with a cytochemical technique in which malachite green was added to glutaraldehyde. This malachite green-glutaraldehyde fixative enhances the visualization of certain phospholipid-related elements. Estrogen induces an affinity of ventricular cardiac lipid inclusions for the cationic dye malachite green. The staining affinity is evidenced only in the estrous female, not in diestrus. In oophorectomized animals, malachite green staining is seen only following estradiol injection, but this effect is blocked by progesterone. In the male, ventricular lipids do not stain, nor do they develop malachite green affinity with estrogen stimulation. These results imply a blockade of the estradiol-mediated dye affinity by progesterone and testosterone. This reinforces the concept of the heart as a target organ for sex steroids and expands the previously described estrogen effects on myocardium.

Animals↗
PubMed1984

Lack of nuclear estrogen binding in benign mesothelioma of the uterus.

Estrogen binding sites in a benign mesothelioma of the uterus was studied by in vitro steroid autoradiography. Unlike normal myometrium, which shows distinct supanuclear localizations of tritiated estradiol, the cellular components of the benign mesothelioma do not appear to have estrogen receptors. Serial sections through the tumor failed to localize sites of estrogen binding. These findings are congruent with an undifferentiated mesothelial organ for the tumor and suggest a lack of response to endogenous sex steroids.

Adult↗
PubMed1984

Effects of 17 beta-estradiol on a permanent line of rat endometrial cells grown in vitro and in vivo.

A permanent line of epithelioid cells derived from a primary culture of rat endometrium by spontaneous transformation in vitro was studied as a model for the actions of estrogen on target cells. The cells contained typical cytosol and nuclear estrogen receptors. Estradiol added in vitro increased the accumulation of cells as much as 60% over controls. Cells injected into ovariectomized athymic mice grew into solid tumors whose incidence was increased by treatment of the animals with estradiol.

Animals↗
PubMed1983

The autoradiographic demonstration of estrogen binding in normal human cervix and vagina during the menstrual cycle, pregnancy, and the menopause.

Using the technique of in vitro steroid autoradiography, the localization and modulation of nuclear estrogen binding sites has been studied in normal human cervix and vagina during the menstrual cycle, pregnancy, and the menopause. Marked differences occur in nuclear estrogen binding between these two organs. Nuclear estrogen binding varies throughout the menstrual cycle in the vaginal epithelium, whereas vaginal stromal cells consistently exhibit nuclear estrogen binding throughout the cycle. In contrast, the cervical squamous and columnar epithelia show much less cyclic variability in nuclear estrogen binding sites. As in the vagina, the cervical stroma consistently binds estrogen. High levels of nuclear estrogen binding sites are found in the vagina of the postmenopausal patient, and lower levels of binding occur postpartum. The implications of these localizations, with special reference to the role of the cervical and vaginal stroma, are discussed.

Adult↗
PubMed1983

Nuclear estrogen binding sites in human endometriosis.

With a technique of in vitro steroid autoradiography, the localization of nuclear estrogen binding sites has been studied in ovarian endometriotic foci from untreated patients in both the follicular and luteal phases of the cycle. Unlike the uterine endometrium, which displays cyclic changes in estrogen binding sites, the endometriotic foci show no such changes in the localization of estrogen binding sites. Throughout the cycle, a marked degree of estrogen binding is present in the stromal cells of the endometriotic foci, while in the uterine endometrium stromal binding sites are seen only during the proliferative phase and not during the secretory phase of the cycle. The glandular epithelium of the endometriotic foci displays a patchy localization of nuclear estrogen binding sites at all stages, while the glandular epithelium of the uterus is strongly positive during the proliferative phase but displays no estrogen binding sites during the secretory phase of the cycle. Thus, the endometriotic foci appear to respond differently to ovarian hormones, both in terms of the modulation of estrogen binding and in terms of glandular histology.

Autoradiography↗
PubMed1983

A rat model for chemotherapy-induced male infertility.

The effects of chronic administration of cyclophosphamide and procarbazine on testicular function in the rat were examined. Cyclophosphamide produced disruption of the normal spermatogenic architecture that was dose and time dependent. Total ablation of the germinal epithelium was not achieved. Procarbazine produced more specific testicular damage. Multiple weekly injections of 100-200 mg/kg of procarbazine caused complete destruction of the spermatogenic cells, with no effect on Sertoli cells. Treated animals remained infertile for more than 4 months and showed no histologic evidence of recovery. This model may be useful for examining possible mechanisms to prevent spermatogenic damage associated with cancer chemotherapy in man.

Animals↗
PubMed1983

Malignant struma ovarii. A case report and literature review.

A woman developed papillary adenocarcinoma in a struma ovarii. The focus of malignancy was within a benign struma found at laparotomy for a pelvic mass. The patient had a previous history of bilateral cystic teratomas. The tumor was composed of papillae of crowded, pleomorphic cells with minimal cytoplasm but a wide range of chromatin patterns. Immunoperoxidase staining for thyroglobulin revealed positive regions in both the benign and malignant areas of tumor. A 15-year review of cystic teratomas in the state of Colorado revealed a frequency of 0.3% for malignant struma ovarii.

Adult↗
PubMed1982

Autoradiographic localization of steroid binding in human tissue labeled in vitro.

A procedure for the rapid preparation of autoradiograms from tissues incubated in vitro with 3H-estradiol is described. Slices of tissue were incubated in culture medium containing 17 nM 3H-estradiol, washed to remove unbound steroid, then processed for thaw-mount autoradiography. Exposure times were generally 3 to 4 weeks. Simultaneous in vitro competition with unlabeled progesterone, dihydrotestosterone, or hydrocortisone had no effect on the distribution or intensity of exposed silver grains, while competition with unlabeled estradiol or moxestrol abolished nuclear localization of silver grains. The exchange of labeled estradiol for bound endogenous estradiol during in vitro incubation of the tissues was demonstrated by a comparison of the pattern of incorporation of 3H-estradiol in tissues previously treated in vivo with unlabeled estradiol versus those not primed. A similar distribution and intensity of silver grains was observed in both the treated and untreated tissue groups. The rationale for the advantages of in vitro steroid autoradiography versus the in vivo technique is discussed.

Adult↗
PubMed1979

Fine structure of fetal human testis and epididymis.

Observations on the structure of the human fetal testis and epididymis at 16 weeks' gestation have been made with the light and electron microscope. The fetal testis is organized into solid cords surrounded by a well-defined cellular investment. The basement membrane is multilaminated with a highly redundant basal lamina. The germ cells rest on thin processes from adjacent Sertoli cells. Intercellular bridges between centrally located germ cells have been observed. The epididymis is remarkably well developed. The tall pseudostratified epithelium lines a discrete duct with a patent lumen. Stereocilia and cilia are seen on apical surface of the principal cells.

Cytoplasm↗
PubMed1978

Malachite green: applications in electron microscopy.

Incorporation of malachite green into a glutaraldehyde fixative results in enhanced staining of a number of cellular elements. Ribosomes and myofilaments exhibit increased electron density, but cell membranes generally are not stained. In certain tissues, lipid inclusions are uniformly and heavily stained. Other populations of lipid droplets exhibit differential affinity for malachite green, facilitating their division into subclasses. In addition to its function as a dye, malachite green has previously been reported to stabilize lipid elements soluble in aqueous glutaraldehyde. Such a component was observed in the stroma of uterine endometrium. The variety of cell components which exhibit increased contrast after preparation with malachite green suggests that this technique may find widespread application in fine structure studies.

Adrenal Cortex↗