Development of macaque secondary follicles exposed to neutral red prior to 3-dimensional culture.

PURPOSE: Neutral red (NR) may assist identification of preantral follicles in pieces of cortical tissue prior to cryopreservation in cancer patients requesting fertility preservation. This study is the first to analyze this effect by follicle growth rate after long-term culture in primates. METHODS: Ovarian cortex was obtained from adult rhesus macaques, was cut into fragments, and was incubated with NR. Secondary follicles were readily visualized following NR staining and then were encapsulated into alginate beads and cultured individually for 4 weeks in αMEM media supplemented with 10 ng/ml FSH at 5% O2. RESULTS: The survival rates of secondary follicles during culture were similar between those derived from control tissue (71 ± 13%) and those treated with NR (68 ± 9%). The proportion of surviving follicles that formed an antrum were also similar in both groups (70 ± 17% control; 48 ± 24% NR-treated). Follicle diameters were not different between control follicles (184 ± 5µm) and those stained with NR (181 ± 7 µm) on the day of isolation. The percentages of surviving follicles within three cohorts based on their diameters at week 4 of culture were similar between the control group and NR-stained tissue group, fast-grow follicles (24 ± 6% vs. 13 ± 10%), slow-grow follicles (66 ± 5% vs. 60 ± 9%), or no-grow (10 ± 9% vs. 27 ± 6%), respectively. There were no differences in follicle diameters between groups during the culture period. Pre-exposure of secondary follicles to NR diminished their capacity to produce both estradiol and androstenedione by week 4 of culture, when follicles are exhibiting an antrum. Inhibitory effects of NR on steroid production by slow-grow follicles was less pronounced. CONCLUSIONS: NR does not affect secondary follicle survival, growth, and antrum formation during long-term culture, but steroid hormone production by fast-grow follicles is compromised. NR can be used as a non-invasive tool for in situ identification of viable secondary follicles in ovarian cortex before tissue cryopreservation without affecting follicle survival and growth in vitro. Whether maturation or developmental competence of oocytes derived from antral follicles in 3D culture that were previously isolated from NR-stained tissue is normal or compromised remains to be determined. Likewise, the functional consequences of pre-exposure to NR prior to ovarian cortical tissue cryopreservation and transplantation are unknown.

Relationship between pelvic floor muscle strength and sexual dysfunction in postmenopausal women: a cross-sectional study.

INTRODUCTION AND HYPOTHESIS: The prevalence of sexual dysfunction in postmenopausal women is high. Theoretically pelvic floor muscle (PFM) strength could influence sexual function, but to date there is scant evidence on this topic. The aim of this study was to evaluate the relationship between PFM strength and sexual function in postmenopausal women. The relationship between reported urinary incontinence (UI) and sexual dysfunction was also investigated. METHODS: This was a cross-sectional study including 113 postmenopausal women. PFM strength was evaluated using vaginal manometry. Sexual function was evaluated using the Female Sexual Function Index (FSFI). A score of ≤26.5 was considered to indicate sexual dysfunction. Urinary incontinence reports were evaluated using the International Consultation on Incontinence Questionnaire-Urinary Incontinence (ICIQ-UI) Short Form. Statistical analysis was performed using Spearman's rank correlation coefficient (ρ), the Mann-Whitney test and 95 % confidence intervals. RESULTS: The median age of the women was 53 years (range 42 - 65 years) and their median body mass index was 27.9 kg/m2 (range 20 - 42 kg/m2). Women without sexual dysfunction showed significantly higher PFM strength (median 41.8, range 11.3 - 94.0 cmH2O) than women with sexual dysfunction (median 30.3, range 3 - 112 cmH2O; p = 0.02). A weak correlation was found between the total FSFI score and the total ICIQ-UI score (ρ = -0.21, p = 0.03). CONCLUSIONS: Postmenopausal women with sexual dysfunction showed lower PFM strength than women without sexual dysfunction. There was a weak correlation between urinary incontinence severity and sexual function.

Sexual function of women practicing sex in nonconventional settings.

The quality of sexual intercourse in the context of conjugal visits by women to their jailed partners is unknown. This study aimed to assess the quality of the sex lives and psychological conditions of women attending conjugal visits with their jailed inmate partners. This controlled study involved 124 women between the ages of 18 to 40 years who engaged in sexual relations with their inmate partners (conjugal visit group) or with their partners at home (control group). Sexual function was assessed using a semi-structured questionnaire and the Female Sexual Function Index, and psychological parameters were evaluated using the Hospital Anxiety and Depression scale. The total Female Sexual Function Index scores was similar in the 2 groups. The percentage of women reporting good quality of the relationship was significantly higher in the conjugal visit group. Also, the Hospital Anxiety and Depression scale scores were higher in the conjugal visit group. Depression was a risk factor for sexual dysfunction and had a negative effect on scores in the desire, excitement, lubrication, and sexual satisfaction domains, whereas anxiety was associated with lower sexual desire scores. A regular + poor quality of the relationship and being religious were factors associated with sexual dysfunction. Sexual practices in jail were not a risk for sexual dysfunction in this sample.

Pelvic floor muscle strength and urinary incontinence in hyperandrogenic women with polycystic ovary syndrome.

INTRODUCTION AND HYPOTHESIS: Hyperandrogenism in women with polycystic ovary syndrome (PCOS) could increase muscle mass and thereby pelvic floor muscle (PFM) strength, reducing the risk of urinary incontinence (UI). The aim of the present study was to assess PFM strength and UI among hyperandrogenic women with PCOS and a control group for comparison. METHODS: This is an observational, cross-sectional, case-control study. Seventy-nine women, aged 18 to 40 years with a mean body mass index (BMI) of 23.4 kg/cm(2) were recruited at the University Hospital: PCOS (n = 36) and control group for comparison (n = 43). All PCOS women had clinical and/or laboratory hyperandrogenism (> 80 ng/dL) and control women had regular menstrual cycles. PFM strength was assessed by vaginal manometry. The International Consultation on Incontinence Questionnaire of Urinary Incontinence Short Form (ICIQ-UI SF) was used to assess UI. Descriptive analysis, analysis of variance (ANOVA) and Fisher's exact test were used for statistical analyses. RESULTS: There was no statistically significant difference in mean PFM strength between the PCOS and the control group: 2.7 cm H2O (95% CI -6.2-11.6) p = 0.55. The prevalence of UI was 18.6% in the control group compared with 0% in the PCOS group p < 0.01. CONCLUSIONS: Women with PCOS showed absence of UI, but PFM strength did not differ from the control group.

Single-cell expression analysis of BMP15 and GDF9 in mature oocytes and BMPR2 in cumulus cells of women with polycystic ovary syndrome undergoing controlled ovarian hyperstimulation.

PURPOSE: To detect expression of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) in oocytes, and their receptor type 2 receptor for BMPs (BMPR2) in cumulus cells in women with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization (IVF), and determine if BMPR2, BMP15, and GDF9 expression correlate with hyperandrogenism in FF of PCOS patients. METHODS: Prospective case-control study. Eighteen MII-oocytes and their respective cumulus cells were obtained from 18 patients with PCOS, and 48 MII-oocytes and cumulus cells (CCs) from 35 controls, both subjected to controlled ovarian hyperstimulation (COH), and follicular fluid (FF) was collected from small (10-14 mm) and large (>18 mm) follicles. RNeasy Micro Kit (Qiagen) was used for RNA extraction and gene expression was quantified in each oocyte individually and in microdissected cumulus cells from cumulus-oocyte complexes retrieved from preovulatory follicles using qRT-PCR. Chemiluminescence and RIA assays were used for hormone assays. RESULTS: BMP15 and GDF9 expression per oocyte was higher among women with PCOS than the control group. A positive correlation was found between BMPR2 transcripts and hyperandrogenism in FF of PCOS patients. Progesterone values in FF were lower in the PCOS group. CONCLUSION: We inferred that BMP15 and GDF9 transcript levels increase in mature PCOS oocytes after COH, and might inhibit the progesterone secretion by follicular cells in PCOS follicles, preventing premature luteinization in cumulus cells. BMPR2 expression in PCOS cumulus cells might be regulated by androgens.

Genital manifestation of graft-vs.-host disease: a series of case reports.

INTRODUCTION: After hematopoietic stem cell transplantation (HSCT), many patients present genital graft-vs.-host disease (GVHD) that can culminate with sexual problems, which are poorly dimensioned. AIM: We hope to draw attention to the need to perform genital biopsy to diagnose genital GVHD, and thus to call attention to the need to incorporate careful attention to sexual health in the treatment of these patients. METHODS: Five allogeneic stem cell transplant recipients complaining of coital pain after HSCT were clinically diagnosed for genital GVHD. Genital biopsies were given for histological analysis, and microphotographs of the corresponding marked field in the slide were taken. Specimens were evaluated by the site pathologist and then sent to a reference pathologist, each blinded to the histological findings. A literature search was performed in PubMed/MEDLINE (1966-2009) for cross-sectional and cohort studies or trials related to genital GVHD. Expert opinions peer reviews and case reports were also considered. MAIN OUTCOME MEASURES: HSCT, genital GVHD, genital biopsy. RESULTS: The biopsy showed evidence of dilated apoptotic cells in the basal layer and detachment of the epithelial lining of the mucosa, hyalinization and thickening of collagen fibers, capillary ectasia, and mononuclear inflammatory infiltrate of the submucosa. Three patients presented vulval lesion such as leucoplasia and ulcer on the large lip. Histological analyses showed evidence of epithelial hyperplasia and influx of inflammatory cells to the epithelial surface, intercellular edema and spongiosis, apoptotic bodies on the basal layer of the epithelium, spongiosis, and nuclear vacuolization. A common treatment based on corticotherapy resulted in complete remission of coetaneous or mucous genital lesions in all five patients. CONCLUSION: Genital biopsy is important to differentially diagnose GVHD and secondary symptoms due to hypoestrogenism. Prevention is the most important step in controlling the evolution GVHD in the vagina to prevent vaginal obstruction and sexual dysfunction.

Sexual dysfunction treatment in a patient with müllerian agenesy submitted to neovaginoplasty: a case report.

Vagina agenesis is a rare entity. Mayer-Rokitansky-Kuster-Hauser syndrome is the most significant cause of vagina agenesis, whereas the second most common cause is complete androgen syndrome. Surgical treatment can propitiate a vaginal reconstruction, but sexual function depends on several factors that affect sexual performance. Many reports focus on the intraoperative and postoperative results and only describe the global approach to these patients, but reports focusing on the management of these patients to enable them to have a normal sexual life are lacking. This case report highlights a multidisciplinary treatment for this kind of morbidity and emphasizes the necessity of incorporating careful attention to sexual health in the treatment of these patients so that they may achieve a good therapeutic response, resulting in a pleasurable sexual life and a good quality of living.

[Concentration of steroid hormones in the follicular fluid of mature and immature ovarian follicles of patients with polycystic ovary syndrome submitted to in vitro fertilization]. / Concentração dos hormônios esteroides no fluido folicular de folículos ovarianos maduros e imaturos de pacientes com síndrome dos ovários policísticos submetidas à fertilização in vitro.

PURPOSE: to evaluate the concentration of steroid hormones in follicular fluid (FF) of small (10-14 mm) and large (> 18 mm) follicles of women with polycystic ovary syndrome (PCOS) submitted to controlled ovarian hyperstimulation (COH) and in vitro fertilization (IVF) cycles. METHODS: a case-control study was conducted on 13 infertile women with PCOS (17 cycles) and 31 infertile women due to male factor - Control Group (31 cycles). FF was aspirated individually and divided into four groups: G1 (FF of small follicles of the Control Group), G2 (FF of small follicles of the PCOS group), G3 (FF of large follicles of the Control Group) and G4 (FF of large follicles of the PCOS group). Estrogen, progesterone and ß-hCG were determined by chemiluminescence, and testosterone and androstenedione by radioimmunoassay. The unpaired t-test was used to compare the hormone determinations in the FF of the PCOS and Control Groups, and the four groups were compared by ANOVA. Fisher's exact test was used to compare the pregnancy rates. RESULTS: the small follicles of the two groups had lower progesterone levels (8,435 ± 3,305 ng/mL) than large follicles (10,280 ± 3,475 ng/mL), p-value <0.01. The progesterone levels of all follicles of group PCOS (8,095 ± 4,151 ng/mL) were lower than Control (9,824 ± 3,128 ng/mL), p-value =0.03. Testosterone differed between G1 (326.6 ± 124.4 ng/dL) and G3 (205.8 ± 98.91 ng/dL), p-value <0.001, and between G3 (205.8 ± 98.91 ng/dL) and G4 (351.10 ± 122.1 ng/dL), p-value <0.001. Small follicles had higher testosterone levels (508.9 ± 266 ng/dL) than large follicles (245.10 ± 123 ng/dL), p-value <0.0001. The pregnancy rates did not differ between the PCOS (5/13, 38.5%) and the Control groups (9/31, 40.9%), p-value =072. CONCLUSIONS: women with PCOS had high testosterone concentrations in the FF, regardless of the stage of follicle development, and reduced progesterone levels, suggesting that paracrine factors may inhibit the secretion of the latter by follicular cells. The pregnancy rates showed that treatment with COH and IVF is a good option for women with infertility secondary to PCOS.

Increased cell proliferation in experimentally induced endometriosis in rabbits.

OBJECTIVE: To characterize the pattern of cell proliferation and apoptosis of eutopic and ectopic endometrium in rabbits after endometrium implantation for the experimental induction of endometriosis. DESIGN: Animal experimental study. SETTING: Sector of experimental surgery. ANIMAL(S): Twenty-female New Zealand rabbits. INTERVENTION(S): All animals underwent laparotomy for endometriosis induction by resection of one uterine horn, isolation of the endometrium, and fixation of tissue segment to the pelvic peritoneum. Two groups of 10 animals were sacrificed 4 and 8 weeks after endometriosis induction. The lesion was excised together with the opposite uterine horn for endometrial gland and stroma determination. MAIN OUTCOME MEASURE(S): Cell proliferation and apoptosis were determined in the eutopic and ectopic endometrium, and the cell proliferation index (CPI) and apoptotic index (AI) were calculated as the number of labeled cells per 1,000 cells. The tissue homeostasis index was the CPI/AI ratio. Glands and stroma were analyzed separately. RESULT(S): The CPI for ectopic tissue was increased compared with eutopic tissue, but there was no difference in the ectopic lesions between 4 and 8 weeks of induction. Considering only the AI, eutopic and ectopic endometrium did not differ after 4 weeks, but differed significantly in glandular tissue after 8 weeks. The tissue homeostasis index revealed cell proliferation in these tissues, with a CPI/AI more than 1. CONCLUSION(S): Ectopic lesions seem to have a higher CPI than eutopic endometrium, with uncontrolled tissue growth occurring in induced endometriotic lesions.

Menopause leading to increased vaginal wall thickness in women with genital prolapse: impact on sexual response.

INTRODUCTION: Hypoestrogenism causes structural changes in the vaginal wall that can lead to sexual dysfunction. A reduction in vaginal wall thickness has been reported to occur after menopause, although without precise morphometry. AIM: To measure vaginal wall thickness in women with genital prolapse in normal and hypoestrogenic conditions and to correlate sexual dysfunction with vaginal wall thickness and estradiol levels. METHODS: Surgical vaginal specimens from 18 normoestrogenic and 13 postmenopausal women submitted to surgery for genital prolapse grades I and II were examined. Patients were evaluated for FSH, estradiol, prolactin, glycemia, and serum TSH levels. For histological analysis, samples were stained with Masson's trichrome and hematoxylin-eosin. Sexual function was assessed by the Golombok-Rust Inventory of Sexual Satisfaction (GRISS). MAIN OUTCOME MEASURES: GRISS questionnaire, histological analysis, morphometric methods, Masson's trichrome. RESULTS: The vaginal wall was thicker in the postmenopausal than premenopausal group (2.72 +/- 0.72 mm and 2.16 +/- 0.43, P = 0.01, and 2.63 +/- 0.71 mm and 2.07 +/- 0.49 mm, P = 0.01, for the anterior and posterior walls, respectively). These thicknesses seem to be due to the muscular layer, which was also thicker in the postmenopausal group (1.54 +/- 0.44 and 1.09 +/- 0.3 mm, P = 0.02, and 1.45 +/- 0.47 and 1.07 +/- 0.44 mm, P = 0.03, for the anterior and posterior wall, respectively). The vaginal epithelium was thinner in the middle segment than in the proximal one in the posterior wall (0.17 +/- 0.07 mm, 0.15 +/- 0.05 mm, 0.24 +/- 0.09 mm, P = 0.02). There was no correlation between coital pain, vaginal wall thickness, and estradiol levels in either group. CONCLUSION: The vaginal wall is thicker after menopause in women with genital prolapse. In this study, vaginal thickness and estrogen levels were not related to sexual dysfunction.

Sexuality during the climacteric period.

BACKGROUND: Cultural, social, physiological and psychological factors may alter the course of sexual function in climacteric women. OBJECTIVE: The objective of the present literature review is to survey the prevalence of sexual dysfunctions in the climacteric and to establish the association between the organic and psychic changes that occur during this phase and sexual dysfunction. We also discuss potential treatments. METHODS: We evaluated the data available in PubMed (1982-2008). For each original article, two reviewers analyzed the data independently and considered a study to be of high quality if it had all three of the following characteristics: prospective design, valid data and adequate sample size. Both reviewers extracted data from each of the 99 studies selected: 34 cross-sectional studies, 25 cohort studies, 9 trials, 31 reviews related to sexuality in pre- and post-menopausal women. RESULTS: Sexual dysfunction among climacteric women is widespread and is associated with bio-psychosocial factors. However, there is not enough evidence to correlate sexual dysfunction with a decrease in estrogen levels and biological aging. A strong association exists between climacteric genital symptoms and coital pain. There is, however, sufficient evidence demonstrating the benefits of local estrogen therapy for patients with genital symptoms. CONCLUSION: A significant decline in sexual function occurs in climacteric women, although it is still unclear whether this is associated with the known decrease in estrogen levels or with aging, or both. Relational factors may interfere with sexual function during this phase. The climacteric genital symptoms improve with estrogen replacement therapy, and positively influence sexual function. Further studies are needed to establish the actual impact of the decrease in estrogen levels and of aging on the sex life of climacteric women.

Uterine ultrasonographic changes during endometriosis treatment: a comparison between levonorgestrel-releasing intrauterine devices and a gonadotropin-releasing hormone agonist.

We compared the effects of levonorgestrel-releasing intrauterine devices (LNG-IUD) and a gonadotropin-releasing hormone agonist (GnRHa) on uterine volume, uterine arteries pulsatility index (PI) and endometrial thickness before and after six months of endometriosis treatment. Sixty women aged 18-40 y were allocated randomly to one of two groups: LNG-IUDs were inserted in 30 women, and GnRHa monthly injections were performed on the other 30. All 60 women were submitted to transvaginal 2-D ultrasound scans on the day that the treatment started and then six months later. Measurements of uterine arteries PI, uterine volume and endometrial thickness were performed at both evaluations. The use of LNG-IUDs significantly decreased endometrial thickness (pre = 6.08 +/- 3.00 mm, post = 2.7 +/- 0.98 mm; mean +/- SD), as did the use of GnRHa (pre = 6.96 +/- 3.82 mm, post = 3.23 +/- 2.32 mm). The uterine volume decreased in the GnRHa group (pre = 86.67 +/- 28.38 cm(3), post = 55.27 +/- 25.52 cm(3)), but not in the LNG-IUD group (pre = 75.77 +/- 20.88 cm(3), post = 75.97 +/- 26.62 cm(3)). Uterine arteries PI increased for both groups; however, the increase was higher in the GnRHa group (0.99 +/- 0.84 vs. 0.38 +/- 0.84, p = 0.007; PI increase in GnRHa and in LNG-IUD groups, respectively). In conclusion, levonorgestrel released directly onto the endometrium by the LNG-IUD induced smaller uterine changes than did the hypoestrogenism induced by GnRHa. Nevertheless, both promoted similar effects on endometrial thickness.

Conservative management of large uterine arteriovenous malformation: case report.

Uterine arteriovenous malformation is a rare condition that may be associated with uterine bleeding. Eventually morphologic aspects of the lesions induce unnecessary aggressive surgery; we report here a case of successful expectant management.

The levonorgestrel-releasing intrauterine device reduces CA-125 serum levels in patients with endometriosis.

This study compared the long-term effects of the levonorgestrel-releasing intrauterine device with those of GnRH agonist administration on serum levels of CA-125 in patients with endometriosis. The levonorgestrel-releasing intrauterine device was found to be as efficient as GnRH agonist in reducing CA-125 serum levels.

Isolated endometrial polypoid sarcoidosis in a post-menopausal patient: case report.

We report here the first case of isolated uterine endometrial sarcoidosis presenting as an asymptomatic polypoid lesion. A 52-year-old patient, who had her last menstrual period 2 years before, was submitted to an office hysteroscopy because of an ultrasonographic finding of endometrial thickening preceding hormonal therapy. A polypoid endometrial lesion removed by surgical hysteroscopy revealed chronic granulomatous endometritis. After excluding other granulomatous diseases, the diagnose of sarcoidosis was corroborated. The finding of polypoid endometrial sarcoidosis expands the differential diagnoses possibilities in case of endometrial polyps.

Use of a levonorgestrel-releasing intrauterine device for the symptomatic treatment of uterine myomas.

OBJECTIVE: To assess the effect of a levonorgestrel-releasing intrauterine device (LNG-IUD) for the treatment of women with uterine myomas with increased bleeding by measuring total uterine volume and by determining patient clinical improvement, uterine artery flow velocity and hemoglobin levels. STUDY DESIGN: Ten patients with a complaint of increased uterine bleeding associated with the presence of uterine myomas were assessed in a descriptive case series and studied before and 6 months after placement of an LNG-IUD. The patients were assessed for intensity of bleeding, uterine volume, Doppler velocimetry of the uterine arteries, hematocrit and hemoglobin. RESULTS: One patient asked to be released from the study 15 days after LNG-IUD insertion, claiming the occurrence of increased bleeding, and 2 patients underwent spontaneous expulsion of the device after 2 and 4 months of follow-up. Of the 7 patients who continued the study, 3 started to present a bleeding pattern of the oligomenorrhea type, and the other 4 experienced amenorrhea. All patients (n = 6) with anemia at the beginning of the study presented normalization of hematocrit and hemoglobin levels after 6 months of treatment (p=0.0003 and p <0.0001, respectively). However, we observed no reduction in uterine volume (p = 0.11) or flow velocity assessed by the uterine artery pulsatility index (right, p=0.17; left, p=0.19) and of the series resistance index (right, p = 0.54; left, p =0.31) using Doppler velocimetry. CONCLUSION: Use of an LNG-IUD may be effective in controlling uterine bleeding secondary to myomatosis even in the absence of reduction of uterine volume.

Insulin resistance in adolescents with menstrual irregularities.

PURPOSE: To assess the presence of insulin resistance as well as the incidence of polycystic ovary syndrome (PCOS) in adolescents with menstrual disorders. METHODS: A case-control study was conducted with 34 adolescents during the period of 2 to 4 years after menarche. The patients were divided into two groups: group I (G I) with 22 patients with menstrual irregularity, and group II (G II) with 12 patients with regular menstrual cycles. Body mass index and Ferriman-Gallway index were calculated for all patients, who also received a pelvic ultrasound. We measured DHEA-S, 17 hydroxyprogesterone, testosterone, TSH, LH, FSH, and prolactin in serum sample and conducted the glucose tolerance test with 75 mg dextrose with measurement of glucose and insulin. RESULTS: Mean +/- SD ovary volume was larger in G I (11.38 +/- 4.06 cm(3)) than in G II (7.72 +/- 5.59 cm(3)); P < 0.05. DHEA-S (G I = 47.23; G II = 38.38 microg/dl) and testosterone (G I = 54.19; G II = 32.53 ng/dl) levels were higher in patients with menstrual irregularity. In G I we detected two patients with diabetes mellitus and one patient with glucose intolerance. Sixteen patients in this group had clinical or hormonal characteristics of PCOS. The mean values of the area under the insulin curve (AUIC) were higher in patients with menstrual irregularities (8,556.52 muIU/mL/2 h) than in controls (5,743.38 microIU/mL/2 h); P < 0.05. CONCLUSIONS: The presence of PCOS was detected in 95% of the adolescents with menstrual irregularity. Patients with menstrual disorders presented higher AUIC values than controls.