Association between cigarette smoking and ovarian reserve among women seeking fertility care.

INTRODUCTION: This study examined the association of smoking with ovarian reserve in a cross-sectional study of 207 women enrolled in the Louisville Tobacco Smoke Exposure, Genetic Susceptibility, and Infertility (LOUSSI) Study and assessed effect modification by NAT2 acetylator phenotype. METHODS: Information on current smoking status was collected using a structured questionnaire and confirmed by cotinine assay. Serum anti-Müllerian hormone (AMH) levels were used to assess ovarian reserve. Diminished ovarian reserve (DOR) was defined as AMH <1ng/mL. Single nucleotide polymorphisms in the NAT2 gene, which metabolizes toxins found in cigarette smoke, were analyzed to determine NAT2 acetylator status. Linear and logistic regression were used to determine the effects of smoking on ovarian reserve and evaluate effect modification by NAT2. Regression analyses were stratified by polycystic ovary syndrome (PCOS) status and adjusted for age. RESULTS: Current smoking status, either passive or active as measured by urinary cotinine assay, was not significantly associated with DOR. For dose-response assessed using self-report, the odds of DOR increased significantly for every additional cigarette currently smoked (Odds ratio, OR:1.08; 95% confidence interval, 95%CI:1.01-1.15); additionally, every 1 pack-year increase in lifetime exposure was associated with an increased odds of DOR among women without PCOS (OR: 1.08 95%CI: 0.99-1.18). These trends appear to be driven by the heavy or long-term smokers. Effect modification by NAT2 genotype was not established. CONCLUSION: A history of heavy smoking may indicate increased risk of diminished ovarian reserve.

Effects of cadmium and high-fat diet on essential metal concentration in the mouse testis.

The effects of exposure to the environmental toxicant cadmium, in combination with obesity, on the metal content in mouse testis were evaluated. Starting in utero and continuing through to 10 or 24 weeks post-weaning, male mice were exposed to cadmium (0, 0.5 or 5 ppm), and fed either a low (LFD) or high fat diet (HFD) post-weaning. Testicular levels of cadmium and essential metals were determined 10 and 24 weeks post-weaning by ICP-MS. Similar to what has been previously observed in the liver, kidney, heart and brain, significant levels of cadmium accumulated in the testis under all exposure conditions. Additionally, HFD-fed animals accumulated more cadmium than did their LFD-treated counterparts. Both treatments affected essential metal homeostasis in the testis. These findings suggest that cadmium and obesity may compromise the reproductive potential in the male mouse by disrupting essential metal levels.

Mitochondria in Ovarian Aging and Reproductive Longevity.

Mitochondrial dysfunction is one of the hallmarks of aging. Consistently mitochondrial DNA (mtDNA) copy number and function decline with age in various tissues. There is increasing evidence to support that mitochondrial dysfunction drives ovarian aging. A decreased mtDNA copy number is also reported during ovarian aging. However, the mitochondrial mechanisms contributing to ovarian aging and infertility are not fully understood. Additionally, investigations into mitochondrial therapies to rejuvenate oocyte quality, select viable embryos and improve mitochondrial function may help enhance fertility or extend reproductive longevity in the future. These therapies include the use of mitochondrial replacement techniques, quantification of mtDNA copy number, and various pharmacologic and lifestyle measures. This review aims to describe the key evidence and current knowledge of the role of mitochondria in ovarian aging and identify the emerging potential options for therapy to extend reproductive longevity and improve fertility.

Levels of angiogenic markers in second-trimester maternal serum from in vitro fertilization pregnancies with oocyte donation.

OBJECTIVE: To determine whether differences exist in angiogenic placental growth factor (PlGF) and antiangiogenic soluble vascular endothelial growth factor receptor 1 (sVEGFR-1; both being early markers of placental ischemic disease) in oocyte-donation (OD) pregnancies, compared with autologous in vitro fertilization (aIVF) and spontaneous pregnancies. DESIGN: Case-control study of residual second-trimester serum samples from women undergoing prenatal screening. SETTING: Academic medical center. PATIENT(S): Fifty-seven OD pregnancies were identified. Each OD pregnancy was matched to two spontaneous pregnancies (n = 114) and one aIVF pregnancy (n = 57). INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): Second-trimester serum PlGF and sVEGFR-1 levels. RESULT(S): sVEGFR-1, PlGF, and unconjugated E2 levels were similar among the three study groups. The ratio of sVEGFR-1 to PlGF was significantly higher in the OD group. Consistently with previous studies, alpha-fetoprotein (AFP) in the OD group was significantly elevated compared with spontaneous pregnancy. Both aIVF and OD groups had greater levels of inhibin A than the spontaneous pregnancy group, and the OD group had significantly higher levels of inhibin A than the aIVF group. hCG levels were significantly elevated in aIVF compared with spontaneous pregnancy; however, levels were not different between aIVF and OD. CONCLUSION(S): Second-trimester serum sVEGFR-1 and PlGF levels were not significantly altered in OD pregnancies. Our data support previous findings that OD pregnancies have uniquely increased second-trimester AFP, hCG, and inhibin A levels compared with aIVF. However, the biologic basis of these marker elevations in OD may not be related to placental angiogenesis.

Minichromosome maintenance complex component 8 and 9 gene expression in the menstrual cycle and unexplained primary ovarian insufficiency.

PURPOSE: DNA repair genes Minichromosome maintenance complex component (MCM) 8 and 9 have been linked with gonadal development, primary ovarian insufficiency (POI), and age at menopause. Our objective was to characterize MCM 8 and 9 gene expression in the menstrual cycle, and to compare MCM 8/9 expression in POI vs normo-ovulatory women. METHODS: Normo-ovulatory controls (n = 11) and unexplained POI subjects (n = 6) were recruited. Controls provided three blood samples within one menstrual cycle: (1) early follicular phase, (2) ovulation, and (3) mid-luteal phase. Six of 11 controls only provided a follicular phase sample. Amenorrheic POI subjects provided a single, random blood sample. MCM8/9 expression in peripheral blood was assessed with qRTPCR. Analyses were performed using delta-Ct measurements; group differences were transformed to a fold change (FC) and confidence interval (CI). Differences across menstrual cycle phases were compared using random effects ANOVA. Two-sample t tests were used to compare two groups. RESULTS: MCM8 expression was significantly lower at ovulation and during the luteal phase, when compared to the follicular phase [FC = 0.69 in the luteal vs follicular phase (p = 0.012, CI = 0.53, 0.90); and 0.65 in the ovulatory vs follicular phase (p = 0.0057, CI = 0.50, 0.85)]. No change in MCM9 expression was noted throughout the menstrual cycle. No significant difference was seen in MCM8/9 expression when comparing POI to control subjects. CONCLUSIONS: Our study showed greater MCM8 expression in the follicular phase of the menstrual cycle, compared to the ovulatory and luteal phases. No cyclic changes were seen with MCM9. Significant differences in MCM8/9 expression were not detected between POI and controls; however, we recommend further investigation with a larger sample population.

Reduced levels of miRNAs 449 and 34 in sperm of mice and men exposed to early life stress.

Exposure of male mice to early life stress alters the levels of specific sperm miRNAs that promote stress-associated behaviors in their offspring. To begin to evaluate whether similar phenomena occur in men, we searched for sperm miRNA changes that occur in both mice and men exposed to early life stressors that have long-lasting effects. For men, we used the Adverse Childhood Experience (ACE) questionnaire. It reveals the degree of abusive and/or dysfunctional family experiences when young, which increases risks of developing future psychological and physical disorders. For male mice, we used adolescent chronic social instability (CSI) stress, which not only enhances sociability defects for >1 year, but also anxiety and defective sociability in female offspring for multiple generations through the male lineage. Here we found a statistically significant inverse correlation between levels of multiple miRNAs of the miR-449/34 family and ACE scores of Caucasian males. Remarkably, we found members of the same sperm miRNA family are also reduced in mice exposed to CSI stress. Thus, future studies should be designed to directly test whether reduced levels of these miRNAs could be used as unbiased indicators of current and/or early life exposure to severe stress. Moreover, after mating stressed male mice, these sperm miRNA reductions persist in both early embryos through at least the morula stage and in sperm of males derived from them, suggesting these miRNA changes contribute to transmission of stress phenotypes across generations. Since offspring of men exposed to early life trauma have elevated risks for psychological disorders, these findings raise the possibility that a portion of this risk may be derived from epigenetic regulation of these sperm miRNAs.

The Value of Anti-Müllerian Hormone in Predicting Clinical Pregnancy After Intrauterine Insemination.

OBJECTIVES: To evaluate the utility of anti-Müllerian hormone (AMH) in predicting clinical pregnancy with intrauterine insemination (IUI) and compare it to other markers of quantitative ovarian reserve. METHODS: Retrospective cohort study of women undergoing natural and stimulated IUI cycles. All patients achieved a clinical pregnancy within three IUI cycles or completed three IUI cycles without pregnancy. Receiver operating curves were generated to determine the ability of AMH, antral follicle count, age, BMI, and day 3 FSH to predict clinical pregnancy with IUI. Characteristics of those with and without pregnancy were compared using Mann-Whitney U, chi-square, and Fisher exact tests. RESULTS: Of 209 women included, 49% achieved clinical pregnancy. Pregnant patients were more likely to have a higher AMH (2.76 vs. 1.55 ng/mL, P = 0.0004). The area under the curve was 0.642 in predicting clinical pregnancy within three IUI cycles using AMH (0.608 if excluding polycystic ovarian syndrome patients); 0.639 using antral follicle count; 0.549 using age; 0.599 using day 3 FSH; and 0.639 using BMI. CONCLUSION: Although serum AMH appears significantly higher in women achieving clinical pregnancy, the predictive value of AMH alone was no better than that for other markers of quantitative ovarian reserve in a patient who clinically qualifies for IUI.

Mitotically Active Leiomyoma in a Woman with Mayer-Rokitansky-Küster-Hauser Syndrome: A Case Report.

BACKGROUND: Cases of women with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome developing leiomyomata are rare. A case with mitotically active leiomyomata has not previously been described to our knowledge. CASE: A 43-year-old woman with MRKH syndrome found to have an incidental pelvic mass on imaging studies underwent a diagnostic laparoscopy, followed by resection of leiomyomata and uterine remnant via mini laparotomy. Histopathology revealed focal infarction associated with a mitotically active area in one of the leiomyomata but with no evidence of marked cytologic atypia or hypercellularity. Focal adenomyosis was also noted. CONCLUSION: Studies have shown that mitotically active smooth cell tumors of the uterus having 5-9 mitoses/10 hpf and no cellular atypia have a metastatic rate too low to be regarded as sarcomas. Although the pathology findings in this case are benign with no need for continued surveillance by gynecologic oncology, regular follow-up with a gynecologist annually may be indicated for early diagnosis of recurrence secondary to the uncommon characteristics of this benign tumor, especially in this rare category of patients with Müllerian agenesis. Mitotically active leiomyomata can occur in patients with Müllerian agenesis, but the likelihood that a pelvic mass in a patient with MRKH syndrome is a sarcoma is extremely low.

Nonsurgical management of primary ovarian pregnancy with transvaginal ultrasound-guided local administration of methotrexate.

Primary ovarian pregnancy is a rare form of ectopic gestation and one that is often diagnosed only at the time of surgery. We report the first case to our knowledge of a live primary ovarian pregnancy diagnosed by transvaginal ultrasonography and successfully treated with transvaginal-guided aspiration and injection of methotrexate. Primary ovarian pregnancy can be diagnosed early in gestation with transvaginal sonography, affording the opportunity to successfully be managed with local administration of methotrexate.

Epigallocatechin-3-gallate, a natural polyphenol, inhibits cell proliferation and induces apoptosis in human ovarian cancer cells.

BACKGROUND: Epigallocatechin-3-gallate (EGCG) is a polyphenol constituent present in green tea previously shown to inhibit cancer growth. However, studies on human ovarian cancer are limited. This study evaluated, the effects of EGCG as a potential anti-proliferative and pro-apoptotic agent in the human ovarian cancer line, SKOV-3. MATERIALS AND METHODS: The MTS assay which measures metabolic activity of cells, bromodeoxyuridine (BrdU) incorporation assay, and flow cytometry were used for the cell proliferation studies and cell morphology, DNA fragmentation analysis and the TUNEL assay for apoptotic effects. RESULTS: The EGCG treated SKOV-3 cells showed inhibition of cell viability and proliferation in a dose-dependent manner. Furthermore, EGCG-mediated SKOV-3 cell growth inhibition was associated with apoptotic changes as evident by cell cycle arrest and accumulation of cells in the apoptotic phase, cell morphological changes, DNA fragmentation and TUNEL-positive apoptotic cells. CONCLUSION: In SKOV-3 cells, EGCG inhibits cell proliferation via DNA synthesis reduction and induces apoptotic cell death via DNA damage, thus elucidating a novel, plausible mechanism of EGCG anti-tumorigenic property.

Intercourse compliance, ovulation, and treatment success in the National Institute of Child Health and Human Development-Reproductive Medicine Network's Pregnancy in Polycystic Ovary Syndrome (PPCOS) Trial.

OBJECTIVE: To investigate the relationship among intercourse compliance, ovulation, and the occurrence of pregnancy in the Reproductive Medicine Network's Pregnancy in Polycystic Ovary Syndrome (RMNPPCOS) Trial. DESIGN: Post hoc data analysis of subjects in the Reproductive Medicine Network PPCOS Trial. SETTING: Academic medical centers. INTERVENTION(S): None. PATIENT(S): Six hundred twenty-six infertile women with polycystic ovary syndrome with a mean age of 28.1+/-4 years and mean body mass index of 35.2+/-8.7 kg/m2. MAIN OUTCOME MEASURE(S): Intercourse compliance, ovulation, and pregnancy. RESULT(S): Data on 2925 cycles were included in the analysis, of which 1340 were ovulatory cycles and 1585 were nonovulatory cycles. The rates of intercourse compliance in the PPCOS trial were similar across all treatment groups at all cycles except cycle 4. Among cycles with known ovulation status, 81.2% of patients were compliant with intercourse instructions. Patients were more intercourse compliant in those cycles during which ovulation occurred (83.2% vs. 79.4%). With regard to ovulatory cycles, there was no difference in the occurrence of pregnancy when comparing intercourse compliant versus intercourse noncompliant cycles. CONCLUSION(S): Intercourse compliance was not associated with the occurrence of pregnancy in ovulatory cycles in the PPCOS Trial. The occurrence of ovulation still remains a critical predictor for the occurrence of pregnancy.

Body mass index and intercourse compliance.

OBJECTIVE: To investigate the relationship between body mass index and intercourse compliance in the Reproductive Medicine Network's Pregnancy in Polycystic Ovary Syndrome (RMN PPCOS) Trial. DESIGN: Post hoc data analysis of subjects in the RMN PPCOS Trial. SETTING: Academic medical centers. INTERVENTION(S): None. PATIENT(S): Six hundred twenty-six infertile women with polycystic ovary syndrome (PCOS) with a mean age of 28.1+/-4 years and mean body mass index (BMI) of 35.2+/-8.7 kg/m2. MAIN OUTCOME MEASURE(S): Intercourse compliance and BMI. RESULT(S): Overall, body mass index was not associated with increased intercourse compliance. However, although patients with BMI>or=35 were less likely to ovulate than patients with BMI<35, they tend to be more compliant with intercourse frequency in ovulatory cycles than patients with BMI<35. CONCLUSION(S): BMI was not associated with intercourse compliance or noncompliance. An elevated BMI in infertile women with PCOS is not associated with poor intercourse compliance.

Eosinophilic perifolliculitis presenting as a painful cystic ovarian mass in a woman with fibromyalgia: a case report.

BACKGROUND: Autoimmune oophoritis is characterized by an ovarian lymphocytic infiltrate and is a rare finding in women with premature ovarian failure. Eosinophilic perifolliculitis is a possible variant of autoimmune oophoritis, of which the pathogenesis and natural history are largely unknown. CASE: A 45-year-old woman, gravida 2, para 2, status post total abdominal hysterectomy, presented to her internist complaining of cyclic, throbbing, right lower quadrant pain. Her past medical history was significant forfibromyalgia. Pelvic ultrasound demonstrated a 2.3-cm, physiologic-appearing right ovarian cyst. Follow-up ultrasound showed a 2.2-cm, complex cyst on the right ovary that increased in size to 4.2 x 3.2 x 3.5 cm on repeat ultrasound 12 weeks later. Exploratory laparotomy and bilateral salpingo-oophorectomy were performed. Pathologic evaluation of the ovaries revealed a 3 x 2 cm regressing corpus luteal cyst with numerous eosinophils, lymphocytes, macrophages and plasma cells, infiltrating the cyst zoall. Serum antiovarian antibodies were positive. CONCLUSION: The patient's pathologic findings are consistent with the rare entity of eosinophilic perifolliculitis. The patient's history offibromyalgia is of particular interest given that both of these diseases may have an autoimmune etiology. Eosinophilic perifolliculitis should be considered in the differential diagnosis of premenopausal and perimenopausal women with pelvic pain and persistent cystic ovarian enlargement.

PGD analysis for aneuploidy in a patient heterozygous for a polymorphism of chromosome 16 (16qh-).

OBJECTIVES: Detection and study of a polymorphism of chromosome 16qh in preimplantational embryos as well as in peripheral blood from the carrier. METHODS: A polymorphism of chromosome 16 (16qh-) was detected in PGD analysis for aneuploidy using a probe for the centromeric region of chromosome 16. The lack of pericentromeric heterochromatin in one of the chromosomes 16 could lead to misdiagnosis in PGD. PGD analysis using telomeric probes for this chromosome was performed to confirm the polymorphism as well as to avoid misdiagnosis in embryo blastomeres. FISH in lymphocyte metaphases was used to detect the carrier. RESULTS: Telomeric probes allowed detection of a polymorphism for the centromeric region of chromosome 16. FISH with centromeric and telomeric probes in lymphocyte metaphases of the parents showed that the patient was the carrier of the polymorphism. CONCLUSION: It is highly recommended that telomeric probes for chromosome 16 be used in preimplantation genetic diagnosis when a high frequency of monosomy for chromosome 16 is observed after regular aneuploidy analysis.

A randomized comparison of group cognitive--behavioral therapy, surface electromyographic biofeedback, and vestibulectomy in the treatment of dyspareunia resulting from vulvar vestibulitis.

This study compared group cognitive-behavioral therapy (12-week trial), surface electromyographic biofeedback (12-week trial), and vestibulectomy in the treatment of dyspareunia resulting from vulvar vestibulitis. Subjects were 78 women randomly assigned to one of three treatment conditions and assessed at pretreatment, posttreatment and 6-month follow-up via gynecological examinations, structured interviews and standard questionnaires pertaining to pain (Pain Rating Index and Sensory scale of the McGill Pain Questionnaire, vestibular pain index, pain during intercourse), sexual function (Sexual History Form, frequency of intercourse, Information subscale of the Derogatis Sexual Functioning Inventory), and psychological adjustment (Brief Symptom Inventory). As compared with pretreatment, study completers of all treatment groups reported statistically significant reductions on pain measures at posttreatment and 6-month follow-up, although the vestibulectomy group was significantly more successful than the two other groups. However, the apparent superiority of vestibulectomy needs to be interpreted with caution since seven women who had been assigned to this condition did not go ahead with the intervention. All three groups significantly improved on measures of psychological adjustment and sexual function from pretreatment to 6-month follow-up. Intent-to-treat analysis supported the general pattern of results of analysis by-treatment-received. Findings suggest that women with dyspareunia can benefit from both medical and behavioral interventions.