Sulfated glycosaminoglycans of periurethral tissue in pre- and postmenopausal women.

OBJECTIVE: The objective was to determine the sulfated glycosaminoglycans (GAGs) of the extracellular matrix (ECM) in pre- and postmenopausal women. STUDY DESIGN: Periurethral tissue was obtained from 44 consecutive women who underwent surgery for urinary incontinence, for pelvic organ prolapse, or for other gynecologic benign conditions. Biopsy specimens were assessed by biochemical methods to characterize and quantify sulfated GAG. Measurements were made of total GAG, chondroitin sulfate, dermatan sulfate and of heparan sulfate. Data were compared using the t-test. RESULTS: Patients were divided into two groups (pre- and postmenopausal groups) and dermatan sulfate was the most predominant glycosaminoglycan. Postmenopausal women had significantly less total sulfated glycosaminoglycans (p<0.01), dermatan sulfate (p<0.01) and chondroitin sulfate (p<0.05) than premenopausal women. We did not observe any differences in heparan sulfate. CONCLUSIONS: Postmenopausal women showed quantitative differences in the biochemical characteristics of the ECM in periurethral tissue by analysis of sulfated GAG.

Expression of vascular endothelial growth factor in the lower urinary tract in rats after castration and estrogen administration.

OBJECTIVE: To evaluate quantitatively, by means of immune histochemistry, the expression of the vascular endothelial growth factor (VEGF) in the bladder, vesicourethral junction, and urethra in normal, castrated adult rats and under estrogen administration. DESIGN: Sixty adult virgin rats (Rattus norvergicus albinus, Rodentia, Mammalia) from the CEDEME-UNIFESP Animal House were used. Rats were divided into three groups. Group I comprised noncastrated rats, group II comprised oophorectomized rats, and group III comprised castrated rats administered 17beta-estradiol in the form of subcutaneous implants at the dose of 0.18 mg/implant for 30 days. After performing standard immunohistochemistry procedures, the intensity of the dark-brown color was used as the cytoplasmic protein expression of VEGF. Cells without this coloration or weakly stained were considered negative. percentile of VEGF expression was obtained by counting 1,000 cells per slide and establishing the ratio between positive and total cells. RESULTS: The VEGF expression was uniform and similar along the urinary tract in group I. After castration, protein expression was almost absent in the bladder and was low in the vesicourethral junction and urethra. With estrogen replacement, very little of the expression was recovered in the bladder, and the reaction became evident in the vesicourethral junction and urethral sections. CONCLUSIONS: The present study implies a potential relationship between VEGF and urinary tract physiology. The results suggest that there are quantitative differences in VEGF expression in these tissues depending on steroid hormone status.

Sulfated glycosaminoglycans of the periurethral tissue in women with and without stress urinary incontinence, according to genital prolapse stage.

OBJECTIVE: The objective was to determine sulfated glycosaminoglycans (GAG) of the extracellular matrix (ECM) in women with and without stress urinary incontinence according to genital prolapse stage. STUDY DESIGN: Periurethral tissue was obtained from 30 women who underwent surgery for urinary incontinence, for pelvic organ prolapse, or for other benign gynecologic conditions. Biopsy specimens were assessed by biochemical methods to characterize and quantify sulfated glycosaminoglycans. Measurements were made of total glycosaminoglycans, chondroitin sulfate, dermatan sulfate, and of heparan sulfate. Data were compared using the t-test. RESULTS: In two groups, dermatan sulfate was the most predominant glycosaminoglycan. Women with stress urinary incontinence had significantly more total sulfated glycosaminoglycans (p<0.05) and dermatan sulfate (p<0.05) than women without stress urinary incontinence. We did not observe any differences in chondroitin sulfate and heparan sulfate. CONCLUSIONS: Women with stress urinary incontinence showed quantitative and qualitative differences in the biochemical characteristics of the extracellular matrix in periurethral tissue by analysis of sulfated glycosaminoglycans, according to genital prolapse stage.

Effects of tamoxifen therapy on the expression of p27 protein in the endometrium of women with primary breast cancer.

The cyclin-dependent kinase inhibitor, p27, has been shown to mediate cell growth arrest thereby significantly reducing the percentage of proliferating cells. It seems that p27 expression is essential for the control of normal endometrial proliferation, and reduced or absent p27 expression may be an important step in endometrial carcinogenesis. Our aim was to demonstrate the effects of tamoxifen therapy on the expression of p27 protein in the endometrium of postmenopausal breast cancer patients. Fifty-three pre- and post-tamoxifen treatment endometrium samples were examined immunohistochemically using p27 antibody. Tamoxifen therapy (20 mg/day) for 60 days increased the expression of p27 protein in the endometrium of postmenopausal breast cancer patients. We conclude that tamoxifen therapy does not seem to be directly involved in the carcinogenesis of endometrial carcinoma since the expression of p27 is not decreased.

Cervical biopsy-based comparison of a new liquid-based thin-layer preparation with conventional Pap smears.

The objective of this study is to compare the diagnostic efficacy of universal collection medium (UCM) liquid-based cytology (LBC) (Digene Corp., MD) and the conventional Pap smear in a comparative study, using histologic results as the gold standard. This was a cross-sectional study. Conventional Pap smears and UCM LBC specimens, obtained from women in a low socioeconomic outpatient population referred to a tertiary center for gynecologic care, were compared. For the purpose of this study, when cervical specimens were collected for cytology, all women underwent colposcopy and biopsy was done if a cervical abnormality was observed. Cytologic evaluation of UCM LBC and conventional Pap smears were carried out separately, masked to the results of the other method. Agreement beyond chance between the two cytologic methods was ascertained by means of the unweighted kappa statistic. Sensitivity, specificity, and predictive values with 95% confidence intervals were calculated for both methods. McNemar's test was used to determine the level of association between the two cytology procedures. A total of 800 women were evaluated. Assessment of the overall agreement between the two cytologic methods yielded a kappa of 0.777 (P < 0.0001). After adjustment for histologic diagnosis, the computed kappa in each stratum was as follows: normal = 0.733; CIN 1 = 0.631; CIN 2/3 = 0.735; cancer = 0.652. The sensitivity and specificity of UCM LBC for detection of cervical intraepithelial lesions and cancer were 75.3% and 86.4%, respectively, not statistically different from the 81.8% and 85.2% seen with the conventional method. This study demonstrates that the UCM LBC method is as accurate as the conventional Pap smear cytology in detecting cervical intraepithelial lesions and cancer even so the UCM samples were systematically prepared from a second sampling of the cervix.

Atypical glandular cells of undetermined significance. Cytologic predictive value for glandular involvement in high grade squamous intraepithelial lesions.

OBJECTIVE: To verify the cytologic predictive value of a diagnosis of atypical glandular cells of undetermined significance (AGUS) in high grade squamous intraepithelial lesions (HSIL) cases. STUDY DESIGN: In a retrospective study, 98 cases of HSIL were reviewed. All patients were referred for colposcopy and directed biopsy to confirm the cytologic diagnoses. Loop excision of the transformation zone was performed to treat clinical lesions. Sensitivity, specificity, positive and negative predictive value were evaluated. Kappa statistics and logistic regression analysis were used to evaluate the findings statistically. RESULTS: By logistic regression analysis, we found that the chance of finding squamous intraepithelial lesions involving glands in AGUS smears was 5.32 times higher than in those with no AGUS. It was 5.74 times higher in cervical intraepithelial neoplasia (CIN) 3 lesions than in CIN 2. CONCLUSION: The cytologic predictive value for HSIL involving glands is statistically significant when specific and objective criteria are used for the AGUS diagnosis.

Role of glutathione S-transferase (GSTM1) gene polymorphism in development of uterine fibroids.

Glutathione S-transferase (GSTM1) plays an important role in the excretion of catechol estrogens and is therefore a candidate marker for fibroids. However, this case-control study demonstrated no association between GSTM1 polymorphism and the risk of leiomyoma in Brazilian women.

Effects of estrogen and estrogen-progestogen therapy on homocysteine levels and their correlation with carotid vascular resistance.

OBJECTIVE: To evaluate the correlation between homocysteine levels and carotid vascular resistance in menopausal women submitted to estrogen and estrogen-progestogen therapy. METHODS: Eighty-six women with a mean age of 52 years were enrolled in a prospective, randomized, double-blind, 6-month study. Patients were allocated to use one of three oral therapies: placebo (n = 26), micronized estradiol 2 mg/day (n = 30) or micronized estradiol 2 mg/day plus norethisterone acetate 1 mg/day (n = 30). Evaluation of homocysteine levels and Doppler sonography of the common carotid artery, used to calculate pulsatility index (PI), were carried out prior to initiating therapy and at the end of the study. The correlation between these two parameters was evaluated using Pearson's coefficient of correlation. RESULTS: There was a significant reduction in homocysteine levels in the groups treated with estrogen alone or estrogen combined with norethisterone. PI was significantly lower only in users of estrogen alone; however, no significant correlation was found between homocysteine measurements and PI. CONCLUSION: No significant correlation was found between homocysteine levels and carotid vascular resistance following hormone therapy.

Metoclopramide-induced hyperprolactinemia affects mouse endometrial morphology.

Information about the effects of pituitary hyperprolactinemia on endometrium, especially in levels coexisting with absence of amenorrhea in women, is scarce. The interference of high prolactin levels on endometrial morphology was thus investigated in young post-pubertal and adult mice rendered hyperprolactinemic by long-term treatment with metoclopramide (MC). No remarkable differences have been noticed upon light microscopy examination of the endometria comparing young to adult cycling MC-treated mice, except on the max/min diameter ratio, which in young animals was lower than in adults (ANOVA, p < 0.01). Both young and adult MC-treated mice presented an increased number of endometrial glands than their respective controls (ANOVA, p < 0.01). However, young MC-treated animals showed the highest values of endometrial thickness index compared to other groups (ANOVA, p < 0.01). Our results indicate that MC-induced hyperprolactinemia causes mouse endometrium proliferation, mainly in young animals.

Glycosaminoglycan profile in bladder and urethra of castrated rats treated with estrogen, progestogen, and raloxifene.

OBJECTIVE: This study was undertaken to evaluate the action of conjugated equine estrogens alone, medroxyprogesterone, the combination of these estrogens with progestogens, and of raloxifene on the glycosaminoglycan profile in the bladder and urethra of adult oophorectomized rats in comparison with noncastrated rats. STUDY DESIGN: Sixty adult rats, of which 50 were submitted to bilateral oophorectomy, were studied. After 4 days, the latter were assigned to five groups of 10 animals each. For 30 consecutive days the following treatments were given: group 1, conjugated equine estrogens; group 2, conjugated equine estrogens combined with medroxyprogesterone acetate; group 3, medroxyprogesterone; group 4, raloxifene; and group 5, placebo. Thereafter the bladders and urethras of the animals were removed, processed to yield a dry powder of which the sulfated glycosaminoglycan content was determined by densitometry after agarose gel electrophoresis and that of hyaluronic acid by a fluorimetric assay. RESULTS: Glycosaminoglycans found in the bladder and urethra were dermatan sulfate, heparan sulfate, and hyaluronic acid. In the bladder, hypoestrogenism or replacement with estrogens led to a lower sulfated glycosaminoglycan content. Replacement with estrogens and/or medroxyprogesterone reverted this effect. Hypoestrogenism decreased the dermatan sulfate/heparan sulfate ratio and reduced hyaluronic acid content. Estrogen therapy reverted this alteration and medroxyprogesterone addition annulled the estrogenic effect. In the urethra, castration did not alter hyaluronic acid content and sulfated glycosaminoglycan content, but raloxifene decreased the latter. CONCLUSION: Castrated rats had a decrease in sulfated glycosaminoglycans and hyaluronic acid content in the bladder. Hormonal replacement altered the quantity and quality of glycosaminoglycans. In the urethra, raloxifene reduced sulfated glycosaminoglycans.

Variacion cuantitativa de los lisosomas en el epitelio de la trompa uterina humana en los periodos menstruales y en la postmenopausia / Quantitative variation of lysosomas in the epitelium of the human uterine tube in the menstrual cycle and in postmenopausal period

Los autores cuantificaron los lisosomas del epitelio de la trompa uterina de mujeres en los períodos menstruales y en la postmenopausia tratadas o no, con estrógenos conjugados durante 21 días. Para esto, fragmentos de la ampolla uterina fueron retirados, fijados y sometidos con el método citoquímico para la detección de la fosfatasa ácida. Los resultados mostraron ser, que el número de lisosomas en la fase proliferativa era significativamente mayor que en la fase secretora o en la postmenopausia. El número de lisosomas en la postmenopausia se muestran significativamente mayor en las mujeres tratadas con estrógenos conjugados. Nuestros resultados indican que los estrogenos son los responsables por el aumento del número de lisosomas