Follicle Depletion Research Articles

Linkage analysis coupled with exome sequencing identified defects in gene ‘X’ causing premature ovarian insufficiency

Background Premature ovarian insufficiency (POI) is defined as a primary ovarian defect characterized by absent menarche (primary amenorrhea) or premature depletion of ovarian follicles before the age of 40 (secondary amenorrhea) with hypergonadotropism and hypoestrogenism. POI results in infertility and lifelong steroid deficiency, and is potentially associated with accelerated health risks such as cardiovascular and neurodegenerative disorders and osteoporosis.

Involvement of a volatile metabolite during phosphoramide mustard-induced ovotoxicity

The finite ovarian follicle reserve can be negatively impacted by exposure to chemicals including the anti-neoplastic agent, cyclophosphamide (CPA). CPA requires bioactivation to phosphoramide mustard (PM) to elicit its therapeutic effects however; in addition to being the tumor-targeting metabolite, PM is also ovotoxic. In addition, PM can break down to a cytotoxic, volatile metabolite, chloroethylaziridine (CEZ). The aim of this study was initially to characterize PM-induced ovotoxicity in growing follicles. Using PND4 Fisher 344 rats, ovaries were cultured for 4days before being exposed once to PM (10 or 30μM). Following eight additional days in culture, relative to control (1% DMSO), PM had no impact on primordial, small primary or large primary follicle number, but both PM concentrations induced secondary follicle depletion (P<0.05). Interestingly, a reduction in follicle number in the control-treated ovaries was observed. Thus, the involvement of a volatile, cytotoxic PM metabolite (VC) in PM-induced ovotoxicity was explored in cultured rat ovaries, with control ovaries physically separated from PM-treated ovaries during culture. Direct PM (60μM) exposure destroyed all stage follicles after 4days (P<0.05). VC from nearby wells depleted primordial follicles after 4days (P<0.05), temporarily reduced secondary follicle number after 2days, and did not impact other stage follicles at any other time point. VC was determined to spontaneously liberate from PM, which could contribute to degradation of PM during storage. Taken together, this study demonstrates that PM and VC are ovotoxicants, with different follicular targets, and that the VC may be a major player during PM-induced ovotoxicity observed in cancer survivors.

Serologic autoimmunologic parameters in women with primary ovarian insufficiency

BackgroundPrimary ovarian insufficiency (POI) is heterogeneous disease defined by amenorrhea or premature depletion of ovarian follicles before the age of 40 years. The etiology of POI is still unclear. The purpose of this study is to evaluate whether women with POI have an elevated serum levels of autoimmunologic parameters.MethodsThe serum from peripheral blood samples which come from 96 POI patients and 100 age-matched health women were analyzed for a series of autoimmune antibodies using protein microarray. The antibodies to double-stranded DNA (ds-DNA), histone (HIS), nuclear ribonucleoprotein (RNP), Sjogren’s syndrome A (SSA/Ro), Sjogren’s syndrome B (SSB/La) and Smith antigen, Jo-1, scleroderma-associated antigen (Scl-70) and centromere (CEN), zona pellucid (ZP), adrenocortical antibodies (ACA),Rheumatoid factor (RF), glomerular basement membrane (GBM), proliferating cell nuclear antigen (PCNA), myeloperoxidase (MPO), proteinase 3 (PR3), thyroid microsomal antibody and antinuclear antibody (ANA)were analyzed.ResultsAmong the 96 women with POI and 100 age-matched health controls, women with POI had significantly elevated circulation levels of Jo-1 and PR3 (p = 0.010 and p = 0.001) whereas circulation levels of ANAs, dsDNA, histone, RNP, Sm, Scl-70, SSA, SSB, CEN, ZP, ACA, RF, GBM, PCNA, MPO and TM antibodies were similar between the two groups.ConclusionsThis study shows that the autoimmune antibodies JO-1 and PR3 were significantly higher in POI women group which suggested that these antibodies may have played special role in POI, but the evaluation of the exact pathways of them remains to be determined.

Acute 7,12-dimethylbenz[a]anthracene exposure causes differential concentration-dependent follicle depletion and gene expression in neonatal rat ovaries

Chronic exposure to the polycyclic aromatic hydrocarbon 7,12-dimethylbenz[a]anthracene (DMBA), generated during combustion of organic matter including cigarette smoke, depletes all ovarian follicle types in the mouse and rat, and in vitro models mimic this effect. To investigate the mechanisms involved in follicular depletion during acute DMBA exposure, two concentrations of DMBA at which follicle depletion has (75nM) and has not (12.5nM) been observed were investigated. Postnatal day four F344 rat ovaries were maintained in culture for four days before a single exposure to vehicle control (1% DMSO; CT) or DMBA (12nM; low-concentration or 75nM; high-concentration). After four or eight additional days of culture, DMBA-induced follicle depletion was evaluated via follicle enumeration. Relative to control, DMBA did not affect follicle numbers after 4days of exposure, but induced large primary follicle loss at both concentrations after 8days; while, the low-concentration DMBA also caused secondary follicle depletion. Neither concentration affected primordial or small primary follicle number. RNA was isolated and quantitative RT-PCR performed prior to follicle loss to measure mRNA levels of genes involved in xenobiotic metabolism (Cyp2e1, Gstmu, Gstpi, Ephx1), autophagy (Atg7, Becn1), oxidative stress response (Sod1, Sod2) and the phosphatidylinositol 3-kinase (PI3K) pathway (Kitlg, cKit, Akt1) 1, 2 and 4days after exposure. With the exception of Atg7 and cKit, DMBA increased (P<0.05) expression of all genes investigated. Also, BECN1 and pAKTThr308 protein levels were increased while cKIT was decreased by DMBA exposure. Taken together, these results suggest an increase in DMBA bioactivation, add to the mechanistic understanding of DMBA-induced ovotoxicity and raise concern regarding female low concentration DMBA exposures.

Validation of the power model of ovarian nongrowing follicle depletion associated with aging in women

To validate recently proposed models of ovarian nongrowing follicle (NGF) decay associated with aging within the context of an independent data set. Prospective investigation. Academic medical center. Normal appearing ovaries collected from 52 women (age 28-51 years) undergoing oophorectomy for benign gynecologic indications. Determining ovarian NGF counts with systematic random sampling rules and a validated fractionator/optical disector technique. The goodness-of-fit of predicted NGF counts based on the power and double Gaussian models and those observed in the validation set was assessed with the calculation of the Akaike information criterion and R(2) values. The goodness-of-fit between observed and expected ovarian NGF counts. The power model was an excellent fit to the observed data. The average difference between the observed and expected NGF count was 0.161 (95% CI, -0.058, 0.327). In the present study population, the power model was a superior fit to the observed data compared with the double Gaussian model. This prospective investigation with an independent set of ovarian NGF counts validates the power model as an excellent characterization of the ovarian NGF decline associated with aging.

Coenzyme Q10 Supplementation and Oocyte Aneuploidy in Women Undergoing IVF-ICSI Treatment.

BACKGROUNDThe age-related reduction in live-birth rate is attributed to a high rate of aneuploidy and follicle depletion. We showed in an animal model that treatment with Coenzyme Q10 (CoQ10) markedly improved reproductive outcome. The aim of this study was to compare the post-meiotic oocyte aneuploidy rate in in vitro fertilization (IVF) and intra cytoplasmic sperm injection (ICSI) patients treated with CoQ10 or placebo.METHODSWe conducted a double blind placebo controlled randomized trial that included IVF–ICSI patients 35–43 years of age. The patients were treated with either 600 mg of CoQ10 or an equivalent number of placebo caps. We compared the post-meiotic aneuploidy rate using polar body biopsy (PBBX) and comparative genomic hybridization (CGH). According to the power calculation, 27 patients were needed for each arm.RESULTSOwing to safety concerns regarding the effects of polar body biopsy on embryo quality and implantation, the study was terminated before reaching the target number of participants. A total of 39 patients were evaluated and randomized (17 CoQ10, 22 placebo), 27 were given the study medication (12 CoQ10, 15 placebo), and 24 completed an IVF–ICSI cycle including PBBX and embryo transfer (10 CoQ10, 14 placebo). Average age, base line follicle stimulating hormone (FSH), peak estradiol and progesterone serum level, as well as the total number of human menopausal gonadotropin (hMG) units—did not differ between the groups. The rate of aneuploidy was 46.5% in the CoQ10 group compared to 62.8% in the control. Clinical pregnancy rate was 33% for the CoQ10 group and 26.7% for the control group.CONCLUSIONNo significant differences in outcome were detected between the CoQ10 and placebo groups. However, the final study was underpowered to detect a difference in the rate of aneuploidy.

Liquid Fertility Management Bait Uptake by Urban Rats within New York City Subway Refuse Rooms

Traditional rodent management tools, such as traps and lethal rodenticides, are acute measures to reduce commensal rodent populations. Given the growing concerns regarding effectiveness, environmental safety, and animal welfare related to these tools, it has become imperative to find new methods. Fertility control is an emerging potential alternative to these tools as a safe, humane and effective method of long-term population management. SenesTech, Inc. has developed a liquid fertility management bait that causes follicle depletion in the ovaries of female Sprague Dawley rats and compromises sperm production in male Sprague Dawley rats in laboratory settings. These studies have shown significant decreases in litter sizes following bait consumption, but acceptance of this bait by wild rodents needed to be confirmed. Bait acceptance was tested within refuse rooms of the New York City subway system. Bait was provided ad libitum for 90 days. Uptake was evaluated by examining, via fluorescence microscopy, the presence of the bait marker rhodamine B, which manifests as fluorescent bands in whiskers. Presence of these bands indicated that 51% of the captured population had consumed the bait, and of these, 58% had taken the bait more than once. These results demonstrate that wild rats will consume a liquid fertility management bait, even in the presence of highly palatable and abundant food within the refuse rooms. This study establishes the successful acceptance of a bait by wild rats. Further investigation is needed to evaluate the effectiveness of this product for the management of urban Norway rat populations.

Effects of Oral Uptake of the Chemosterilant 4-Vinylcyclohexene Diepoxide in Wild House Mice, Mus domesticus

The chemical 4-vinylcylcohexene diepoxide (VCD) induces depletion of primordial follicles in the ovaries of laboratory mice after intraperitoneal treatment and leads to infertility. We assessed the oral uptake of a range of doses of VCD by wild house mice, Mus domesticus, in captivity. In Experiment 1, female mice (n = 8 per group) were presented with a daily dose of liquid emulsion containing different concentrations of VCD in a volume equivalent to 10% body weight for 5 days in the presence of ad libitum mouse chow and water. Body weight, body condition, and general health status of the mice were assessed daily. Mice were killed 17 days after cessation of dosing. Internal organs were examined for normality and ovaries were fixed for analysis of primordial follicle populations. Over the 5-day treatment period, 95-100% of mice consumed all their daily dose of VCD emulsion. There were no effects on body weight, general activity, or alertness of the mice. Primordial follicle populations showed dose dependent but variable levels of depletion. In Experiment 2, the highest, most palatable VCD dose (300 mg VCD/kg) or a control dose was presented to mice (n = 24-30 per group), daily for 5 days. At 10 days post treatment, males were introduced for 3 breeding rounds. The numbers of pups/litter and proportions of females producing litters for each breeding round was not different between groups. In Experiment 3, male mice were presented with this same VCD dose; assessment of their reproductive status at 7 and 15 days post treatment indicated no treatment effect on testis and epididymis weights. These experiments indicate that while some follicle depletion was achieved in females with the orally delivered VCD dose, this was insufficient to affect their fertility. Further studies will require presentation of higher doses of VCD and/or extended presentation of this chemosterilant.

Development of a Mouse Model of Menopausal Ovarian Cancer

Despite significant understanding of the genetic mutations involved in ovarian epithelial cancer and advances in genomic approaches for expression and mutation profiling of tumor tissues, several key questions in ovarian cancer biology remain enigmatic: the mechanism for the well-established impact of reproductive factors on ovarian cancer risk remains obscure; cell of origin of ovarian cancer continue to be debated; and the precursor lesion, sequence, or events in progression remain to be defined. Suitable mouse models should complement the analysis of human tumor tissues and may provide clues to these questions currently perplexing ovarian cancer biology. A potentially useful model is the germ cell-deficient Wv (white spotting variant) mutant mouse line, which may be used to study the impact of menopausal physiology on the increased risk of ovarian cancer. The Wv mice harbor a point mutation in c-Kit that reduces the receptor tyrosine kinase activity to about 1–5% (it is not a null mutation). Homozygous Wv mutant females have a reduced ovarian germ cell reservoir at birth and the follicles are rapidly depleted upon reaching reproductive maturity, but other biological phenotypes are minimal and the mice have a normal life span. The loss of ovarian function precipitates changes in hormonal and metabolic activity that model features of menopause in humans. As a consequence of follicle depletion, the Wv ovaries develop ovarian tubular adenomas, a benign epithelial tumor corresponding to surface epithelial invaginations and papillomatosis that mark human ovarian aging. Ongoing work will test the possibility of converting the benign epithelial tubular adenomas into neoplastic tumors by addition of an oncogenic mutation, such as of Tp53, to model the genotype and biology of serous ovarian cancer. Model based on the Wv mice may have the potential to gain biological and etiological insights into ovarian cancer development and prevention.

112 OVARIAN FOLLICLE RESERVE IN NELLORE AND ANGUS HEIFERS WITH LOW AND HIGH FOLLICLE COUNT

Cattle from Bos indicus breeds recruit and develop more follicles per oestrous cycle compared with cattle from Bos taurus breeds. The number of antral follicles in the ovaries is a decisive factor for the success of reproductive technologies such as ovum pick-up and embryo transfer. The reasons why indicine cattle recruit and develop more follicles are not known. Our objectives were to evaluate the ovarian reserve of Nellore and Angus heifers and determine at which stage of follicle development Nellor ovarian reserve differs from Angus ovarian reserve. To determine if animals were in the low or high follicle count group (LFC or HFC), antral follicles (≥3 mm) were scanned with an ultrasound device (US; Mindray Vet DPS 2200, Sao Paulo, Brazil) equipped with a 7.5-MHz probe Day 1 after ovulation (Day 0) for 3 consecutive oestrous cycles. Eighteen Nellore heifers (11 LFC and 7 HFC) and 25 Angus heifers (12 LFC and 13 HFC) were used in the experiment. Oestrous cycles were synchronized with 2 doses of PGF2α ± 11 days apart. Animals were slaughtered ~24 h after ovulation of the third oestrous cycle. Ovulation was determined by oestrous observation, absence of the dominant follicle on the ultrasound, and presence of the corpus luteum after slaughter. One ovary from each animal was used for histology. The ovary was sliced longitudinally in 6 parts, fixed in 10% paraformaldehyde for 24 h, and stored in 70% ethanol until processing. Two longitudinal slices were blocked in paraffin. One section (8 μm thick) was stained after every 50 sections, totaling 10 sections per longitudinal slice of ovarian tissue. Sections were stained using eosin and hematoxylin. Follicles were counted and classified under a stereo microscope with a 40× objective; only follicles with a visible nucleus were counted. Follicles were classified as primordial, primary, and secondary. The area of each section was measured using the hand tool on the ImageJ software. Area measurements were expressed as pixels squared. The density of each follicle classification was calculated as follicle/log10 [pixel2]. Data were analysed using PROC GLM procedure of SAS 9.2 considering the effects of breed and group (LFC or HFC). Angus heifers had more primordial follicles (P &lt; 0.02) when compared with Nellore heifers, but there was no difference within groups. There was no difference in the number of primary follicles within breeds or groups. Secondary follicle numbers tended to be higher (P = 0.09) in Nellore heifers when compared with Angus heifers and was higher within groups (P &lt; 0.02), with HFC Angus and Nellore heifers having more follicles than LFC heifers. It is possible that HFC heifers have a mechanism that is developing more follicles beginning at the secondary stage, especially Nellore heifers. The development of more secondary follicles may contribute to the depletion of primordial follicles observed in Nellore heifers. This research is funded by Sao Paulo Research Foundation (FAPESP) grant #2011/50964-0. Scholarships for Favoureto, Queiroz, Loureiro, Ereno, and Pupulin are also from FAPESP.

ACCELERATED FOLLICLE DEPLETION IN VITRO AND IN VIVO IN SPRAGUE-DAWLEY RATS USING THE COMBINATION OF 4-VINYLCYCLOHEXENE DIEPOXIDE AND TRIPTOLIDE

Rodent pests cause major damage to the world's agricultural crops and food stores. Rodenticides used since World War II did not lead to sustained reduction of rodent populations, and so fertility control is becoming attractive because rats reproduce with great efficiency. Chemical acceleration of ovarian failure via oral dosing also would improve management of rat pest populations. The chemical 4-vinylcyclohexene diepoxide (VCD) is orally efficacious, causing depletion of nonregenerating primordial ovarian follicles of Sprague-Dawley rats. However, to cause rapid reduction in pups in the first breeding cycle after dosing, all stages of ovarian follicle development must be targeted. To achieve this goal, the Chinese herb triptolide was tested because it can precipitate apoptosis and deplete growing follicles. The impact of triptolide was tested in cultured postnatal day 4 Sprague-Dawley rat pup ovaries. Triptolide at 5 nM caused 100% primordial, primary, and secondary follicle depletion after 8 days of culture, compared to 38% follicle depletion caused by VCD at 30 microM. Next, a palatable rat bait was developed, containing 1% VCD with increasing concentrations of triptolide at 25, 50, and 100 microg/kg body weight. Rats ate an average 3-6% of their body weight/day over 15 feeding days. Two days after the end of baiting, rats were euthanized to conduct necropsies and collect ovaries to count all follicular stages and corpora lutea. At 50 microg triptolide/kg body weight, there was significant reduction of all follicular stages; primordial follicles were 50% lower, secondary follicles were 64% lower, antral follicles were 80% lower, and there were no corpora lutea. These results suggest that combining VCD and triptolide in an oral bait leads to significantly compromised rat ovarian function and reduced ovulations, and is likely to reduce pup production.

Cyclooxygenase-1 Inhibition Prolongs Postnatal Ovarian Follicle Lifespan in Mice1

Menopause is the permanent cessation of menstruation that results from depletion of ovarian germ cells and follicles. Although most animals experience reproductive senescence, the mechanisms differ from that in women, who may live more than one-third of their lives after menopause and consequently face the risk of a number of menopause-associated health problems. Understanding factors that influence ovarian aging may provide strategies to delay or alleviate physiological alterations that take place in postmenopausal women. The germ cell-deficient Wv mice recapitulate follicle loss, prolong postreproductive lifespan, and model many physiological changes that take place in postmenopausal women. Here, using genetic and pharmacological approaches, we found that inhibition of cyclooxygenase-1 but not cyclooxygenase-2 in Wv mice delays germ cell depletion and preserves ovarian follicles. Cyclooxygenase-1 inhibition slows down follicle maturation at the conversion of primary to secondary follicles and prolongs postnatal ovarian follicle lifespan. The current study suggests that inhibition of cyclooxygenase-1 may be able to delay ovarian aging and modulate menopausal timing.

Validation of the power model of ovarian non-growing follicle depletion associated with aging

The primary determinant of reproductive aging in women is the progressive decline in ovarian non-growing follicles (NGFs). This pool of NGFs, inclusive of the ovarian primordial, intermediate and primary follicles, constitutes the ovarian reserve. Ultimately the depletion of the NGF pool results in menopause. We have previously characterized the decline in NGFs associated with aging, determining that a power model taking the form y = axb + c, where a, b, and c are constants and y = NGF count at age x, best characterizes the decline in the log-transformed NGF count associated with increasing age. This investigation was aimed at validating the power model of ovarian NGF decay within the context of an independent data set. Prospective cross-sectional study, university setting. Normal ovaries were collected from 55 women (age 28-52 years) undergoing oophorectomy for benign indications. Using systematic random sampling rules and a validated fractionator/optical disector technique, ovarian NGF counts were determined. The goodness-of-fit of predicted NGF counts based on the power model {Log (NGF count) = (-0.00019)*(age in years)2.452 + 5.717} and those observed in the validation set was assessed with a chi-square goodness of fit test, where a p-value > 0.05 is considered a good fit for the model. The power model was an excellent fit to the observed data (p = 0.97). The average difference between observed and expected NGF count was 0.17 (95% CI -0.02,0.345). This difference would translate into a discrepancy of ∼506 NGFs at the predicted menopause threshold of ∼758 NGFs. This investigation, with an independent set of ovarian NGF counts, validates the power model of ovarian NGF depletion associated with aging. These findings confirm the utility of the power model as the best available characterization of the decline in ovarian NGFs from birth to menopause.

Mutations ineIF4ENIF1Are Associated With Primary Ovarian Insufficiency

Primary ovarian insufficiency (POI), or premature ovarian failure, results from ovarian follicle depletion with a consequent elevation of FSH levels before age 40 years. We identified a family in which 9 women in 3 consecutive generations developed menopause at approximately age 30 years. We hypothesized a genetic cause with a dominant mode of inheritance. This was a family-based genetic study and a replicate group of women with POI. The study was conducted at an academic medical center. Seven affected women and an obligate carrier and 7 unaffected family members were genotyped. The genes of interest were also sequenced in 38 unrelated women with POI. The DNA from 7 family members was subjected to whole-exome sequencing. The genotypes of interest were confirmed and genotypes of additional family members and unrelated women with POI were determined using Sanger sequencing. A high-impact, deleterious variant that segregated appropriately with POI in the family was required. A heterozygous stop codon (Ser429X) was identified in the eukaryotic translation initiation factor 4E nuclear import factor 1 (eIF4ENIF1) in the proband and all affected women but not in the unaffected family members. The chance that such a high-impact, deleterious variant would segregate appropriately among the affected and unaffected relatives by chance is very low (P < .05). There were no additional mutations identified in eIF4ENIF1 or eIF4E in 38 unrelated women with POI. Data demonstrate a new gene associated with dominantly inherited POI. These results highlight the importance of translation initiation factors and their regulators in ovarian function.

Cyclophosphamide Exposure in Pediatric Systemic Lupus Erythematosus Is Associated with Reduced Serum Anti-Müllerian Hormone Levels

To the Editor: The reproductive risk to girls with pediatric systemic lupus erythematosus (pSLE) of varying disease severity and medication exposure is not well established. Because 15%–20% of patients with SLE are diagnosed before age 19 years1, many young women with pSLE will be affected by disease complications throughout their reproductive years. Although infertility in SLE has been attributed mainly to use of cyclophosphamide (CYC)2,3, further studies are needed to clarify the relationship between disease severity, medication use, and ovarian dysfunction. Because the incidence of premature primary ovarian insufficiency (POI) after CYC exposure in patients with pSLE < 21 years of age is estimated at 0–11%3, biomarkers for morbidities, such as infertility, are important because they may change current treatment strategies. Anti-Mullerian hormone (AMH) is produced by granulosa cells of preantral and small antral follicles and has been identified as a sensitive biomarker of ovarian reserve. AMH concentrations are relatively stable throughout the menstrual cycle, are unaffected by hormonal contraceptives, and decline with advancing age, as does ovarian reserve/function4,5. Low to undetectable levels of AMH are found in women ages 25–46 years within 5 years of their final menstrual period, in cancer survivors exposed to chemotherapy and/or radiation therapy-induced follicle depletion, and in women with POI … Address correspondence to Dr. N.C. Douglas, Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, New York Presbyterian Hospital–Columbia University Medical Center, 622 West 168th Street, PH 16-64, New York, NY 10032, USA. E-mail: nd2058{at}columbia.edu

Scrambled and fried: Cigarette smoke exposure causes antral follicle destruction and oocyte dysfunction through oxidative stress

Cigarette smoke is a reproductive hazard associated with pre-mature reproductive senescence and reduced clinical pregnancy rates in female smokers. Despite an increased awareness of the adverse effects of cigarette smoke exposure on systemic health, many women remain unaware of the adverse effects of cigarette smoke on female fertility. This issue is compounded by our limited understanding of the molecular mechanisms behind cigarette smoke induced infertility. In this study we used a direct nasal exposure mouse model of cigarette smoke-induced chronic obstructive pulmonary disease to characterise mechanisms of cigarette-smoke induced ovotoxicity. Cigarette smoke exposure caused increased levels of primordial follicle depletion, antral follicle oocyte apoptosis and oxidative stress in exposed ovaries, resulting in fewer follicles available for ovulation. Evidence of oxidative stress also persisted in ovulated oocytes which escaped destruction, with increased levels of mitochondrial ROS and lipid peroxidation resulting in reduced fertilisation potential. Microarray analysis of ovarian tissue correlated these insults with a complex mechanism of ovotoxicity involving genes associated with detoxification, inflammation, follicular activation, immune cell mediated apoptosis and membrane organisation. In particular, the phase I detoxifying enzyme cyp2e1 was found to be significantly up-regulated in developing oocytes; an enzyme known to cause molecular bioactivation resulting in oxidative stress. Our results provide a preliminary model of cigarette smoke induced sub-fertility through cyp2e1 bioactivation and oxidative stress, resulting in developing follicle depletion and oocyte dysfunction.

The Relationship Between Anti-Müllerian Hormone in Women Receiving Fertility Assessments and Age at Menopause in Subfertile Women: Evidence From Large Population Studies

Context: Anti-Mullerian hormone (AMH) concentration reflects ovarian aging and is argued to be a useful predictor of age at menopause (AMP). It is hypothesized that AMH falling below a critical threshold corresponds to follicle depletion, which results in menopause. With this threshold, theoretical predictions of AMP can be made. Comparisons of such predictions with observed AMP from population studies support the role for AMH as a forecaster of menopause. Objective: The objective of the study was to investigate whether previous relationships between AMH and AMP are valid using a much larger data set. Setting: AMH was measured in 27 563 women attending fertility clinics. Study Design: From these data a model of age-related AMH change was constructed using a robust regression analysis. Data on AMP from subfertile women were obtained from the population-based Prospect-European Prospective Investigation into Cancer and Nutrition (Prospect- EPIC) cohort (n � 2249). By constructing a probability distribution of age at which AMH falls below a critical threshold and fitting this to Prospect-EPIC menopausal age data using maximum likelihood, such a threshold was estimated. Main Outcome: The main outcome was conformity between observed and predicted AMP. Results: To get a distribution of AMH-predicted AMP that fit the Prospect-EPIC data, we found the critical AMH threshold should vary among women in such a way that women with low age-specific AMH would have lower thresholds, whereas women with high age-specific AMH would have higher thresholds (mean 0.075 ng/mL; interquartile range 0.038–0.15 ng/mL). Such a varying AMH threshold for menopause is a novel and biologically plausible finding. AMH became undetectable (�0.2 ng/mL) approximately 5 years before the occurrence of menopause, in line with a previous report. Conclusions: The conformity of the observed and predicted distributions of AMP supports the hypothesis that declining population averages of AMH are associated with menopause, making AMH an excellent candidate biomarker for AMP prediction. Further research will help establish the accuracy of AMH levels to predict AMP within individuals.

Differentiation of rat iPS cells and ES cells into granulosa cell-like cells &amp;lt;italic&amp;gt;in vitro&amp;lt;/italic&amp;gt;

Premature ovarian failure (POF) is an ovarian defect characterized by the premature depletion of ovarian follicles before 40 years of age, representing one major cause of female infertility. Stem cells provide the possibility of a potential treatment for POF. In this study, rat embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) were co-cultured with granulosa cells (GCs) to differentiate to GC-like cells. The level of estradiol (E2) analyzed by radioimmunoassay showed that the E2 concentration of the culture supernatant of co-cultured rat iPSCs and ESCs increased in a time-dependent manner, compared with the GCs group that has an opposite trend. The expression of follicle-stimulating hormone receptor (FSHR) was confirmed by immunostaining. These results indicated that rat iPSCs and ESCs were effectively induced to GC-like cells through indirect cell-to-cell contact. Real-time polymerase chain reaction was performed to analyze the expression level of marker genes in POF, including BMP15, FMR1, FSHR, INHA, AMH, NOBOX, FOXO3, EIF2B, FIGLA, and GDF9. The BMP15, FSHR, INHA, AMH, NOBOX, and GDF9 genes were significantly up-regulated in iPSCs and ESCs co-cultured with GCs in comparison with cells that were not co-cultured. Thus, here we demonstrated an available method to differentiate rat iPSCs and ESCs into GC-like cells in vitro for the possible cell therapy of POF.

Morpho-physiological features associated with menopause: recent knowledge and areas for future work.

Menopause is the permanent cessation of menstruation resulting from loss of ovarian follicular activity which happens as a result of depletion of primary follicles which is basically an aging effect. Depletion of ovarian follicles is reflected as declined production of oestradiol which is currently known to be central to the morphologic and physiological changes that happen during the climacteric, menopause and post menopause periods. The cessation of oestradiol production is much more pronounced in tissues with oestrogen receptors such as bones, brain, blood vessels, central nervous system and the skin. But generally little is known on the subject and in particular the bioactive substances involved in the process such that there are some symptoms that menopause women experience which not only defy clinicians but also challenge the management of the condition. This article is presented to shade light to what is currently known, what is not known and stimulate future research which may reveal more understanding and advance our knowledge on management of women throughout the climacteric and menopausal periods.

Peripheral WBC Telomere Length as a Marker of Reproductive Age in Women

Reproductive aging is a continuous process that begins many years prior to menopause. The primary mechanism of reproductive aging is through the process of ovarian primordial follicle depletion, a process that exhibits considerable variation between women. The relationship between somatic and reproductive aging in women has been controversial, with some investigations suggesting a correlation between the two processes, whereas others have suggested no relationship. Peripheral lymphocyte telomere length is widely considered to be a marker of somatic aging.