Fertility Problems In Women Research Articles

Induction of apoptosis by Eleutherine bulbosa (Mill.) Urb. bulb extracted under optimised extraction condition on human retinoblastoma cancer cells (WERI-Rb-1)

Ethnopharmacological relevanceThe bulb of Eleutherine bulbosa (Mill.) Urb. is an indigenous medicinal plant traditionally used among Dayak people for the management of diabetes, breast cancer, hypertension, stroke, and fertility problems in women. The bulb has been reported with a potent cytotoxic potential but with limited underlying mechanisms. Aim of the studyThis study aimed to investigate the cytotoxic properties of E. bulbosa ethanolic bulb extracted under optimised extraction condition on retinoblastoma cancer cells (WERI-Rb-1) through in vitro cell culture bioassays. The optimised extraction condition has been determined in the previous reports. Materials and methodsCytotoxic assay was analysed through MTT assay. Comparison between non-optimised and optimised extraction condition from E. bulbosa ethanolic bulb extract was evaluated. Morphological assessment of apoptotic cells was conducted through acridine orange propidium iodide (AOPI) staining using fluorescence microscopy. Apoptosis assay was carried out through Annexin V-FITC and cell cycle analysis through PI staining. The effect of varying concentrations (IC25, IC50, IC75) of the optimised E. bulbosa ethanolic bulb extract was observed. The mRNA expression was also conducted to confirm the underlying mechanism. ResultsThe optimised E. bulbosa ethanolic bulb extract markedly suppressed the proliferation of retinoblastoma cancer cells significantly with an IC50 value of 15.7 μg/mL as compared to non-optimised extract (p < 0.01). Fluorescence microscopy revealed that retinoblastoma cancer cells manifested early features of apoptosis-like membrane blebbing, chromatin condensation and formation of apoptotic bodies in a dose-dependent manner. The number of apoptotic cells were greatly observed in early and late apoptosis through Annexin V-FITC and the extract also induced cell arrestment as compared to the untreated group. The apoptosis was confirmed with the upregulation of Bax, Bad, p53, Caspase 3, Caspase 8, and Caspase 9 genes meanwhile, Bcl-2, BcL-xL, Nrf-2, and HO-1 genes were downregulated. ConclusionThe optimised E. bulbosa ethanolic bulb extract induced a significant cell death and cell cycle arrestment on retinoblastoma cancer cells. It could be suggested that the induction of apoptosis in retinoblastoma cancer cells may be due to the synergistic effect of the bioactive compounds extracted under optimised extraction condition. Our findings indicated that E. bulbosa bulb could be promising chemotherapeutic potential to treat retinoblastoma cancer cells.

Assisted Reproductive Technologies for Women with Polycystic Ovarian Syndrome

Polycystic ovarian syndrome (PCOS) is the most common cause of anovulatory fertility problems in women of reproductive age, with a prevalence as high as 40% to 75%. There are different treatment modalities for increasing the chance of pregnancy in PCOS patients. Since most of the PCOS patients suffer from ovulatory problems, therapeutic efforts are focused mainly towards restoring normal ovulation. However, even after pharmacological interventions to ameliorate ovulatory irregularities, women with polycystic ovary are still at high risk of fetal loss. Thus, assisted reproductive techniques (ARTs) have become popular approaches which significantly increase the chances of successful pregnancy in these women, bypassing the conditions of PCOS. The present mini-review ‘pinpoints’ on the suitability of ARTs for PCOS patients who fail to ovulate following the conventional infertility treatment. The article also briefly explains how intracytoplasmic sperm injection (ICSI), the most effectiveART, correlates with better biological parameters, fertilization rate and better quality of embryos in PCOS women.

Subcutaneous grafting of ovaries from newborn rabbits into adult ovariectomized mice does not result in normal folliculogenesis

About 10% (6.1 million) of women in the United States ages 15–44 have difficulty becoming pregnant or staying pregnant. About 1/3 are fertility problems in women. Abnormal ovarian function is a major contributor. A greater understanding of ovarian function and especially follicular development would improve treatment. Follicles form when germ cells arrive in the primitive sex cords, which are derived from the coelomic epithelium. In most species, these cords fragment and the germ cells become covered by precursors of the granulosa cells. Thus primordial follicles form. Pools of these follicles are eventually recruited and undergo maturation; a series of functional and morphological transformations that lead to ovulatory follicles. In species like humans, cattle and pigs, follicles form during the embryonic phase of gestation, while maturation takes place after birth. In contrast, both phases occur postnatally in rabbits and makes them an excellent model to study folliculogenesis. Laboratory studies using non‐rodent species are logistically more difficult and even prohibitive. One alternative is ovarian grafting into immunocompromised mice. When adult ovarian tissue is transplanted from large animals into mice, some primordial follicles survive and undergo maturation. We investigated the rabbit model by transplantation of immature rabbit ovaries under the dorsal skin of adult ovariectomized NCr N ude mice (1 ovary / mouse). At 4 – 5 weeks, growth of the grafted ovaries was evident. Macroscopically, 50% of the grafts (5/10) had one or two follicle‐like structures on their surface. Histologically the structures were cysts lined by cuboidal cells and contained cells with the germ cell phenotype. In conclusion, ovaries grafted subcutaneously from newborn rabbits into ovariectomized adult mice developed cystic primordial follicles. Normal follicle formation and maturation did not occur.Support or Funding InformationMidwestern University intramural funds

Serum Vitamin D Levels in Fertile and Infertile Women with Polycystic Ovary Syndrome.

Introduction In polycystic ovary syndrome, serum vitamin D levels are known to correlate with metabolic conditions such as diabetes mellitus, metabolic syndrome and cardiovascular disease. However, there are not enough studies showing such a relationship with female fertility. We aimed to compare serum vitamin D levels in fertile and infertile women with polycystic ovary syndrome to evaluate whether vitamin D may play a role in the pathogenesis of fertility problems in women with polycystic ovary syndrome. Materials and Methods 274 infertile and 111 fertile women with polycystic ovary syndrome were included in this retrospective study. Infertile and fertile groups were matched by age, body mass index and homeostasis model assessment of insulin resistance. Anthropometric, clinical and laboratory characteristics of the women were recorded. Serum 25(OH)D 3 levels were used to assess serum vitamin D levels. Results No significant differences were detected between groups in terms of anthropometric, clinical and laboratory features except for serum 25(OH)D 3 levels and the incidence of vitamin D deficiency. Vitamin D levels were significantly lower and vitamin D deficiency was more common in the infertile group compared to the fertile group. When the groups were stratified into obese/non-obese or insulin resistance positive/negative, infertile obese and infertile insulin resistance-positive women had the lowest serum 25(OH)D 3 levels. Conclusion Serum vitamin D levels are lower in infertile women with polycystic ovary syndrome compared to fertile women. When insulin resistance or obesity was present, vitamin D levels were reduced further. Thus, in polycystic ovary syndrome, lower vitamin D levels may play a role in the pathogenesis of fertility problems.

Etiology of Central (Hypogonadotropic) Female Hypogonadism: Genetic, Organic and Functional Causes

Central (hypogonadotropic) hypogonadism (CH) can be mentioned among the causes of menstrual and fertility problems in women. This clinical syndrome is characterized by decreased sex steroid secretion due to the impairment of basal and/or impulse secretion of gonadotropins. It can be congenital or acquired, organic or functional (i.e. without visible anatomic damages of the hypothalamo-hypophyseal area), isolated or combined with other pituitary hormone deficits (hypopituitarism). All causes of central female hypogonadism can be conditionally divided into some subgroups: genetic factors (lead to congenital CH), organic factors (lead to acquired CH), and idiopathic (lead functional CH). Congenital CH can be caused by genes that control ontogenesis and migration of GnRH neurons, regulate secretion of GnRH and provide its’ biological effects on gonadotropins. The potential organic causes of CH include pituitary tumors and some other types of tumors with similar localization, consequences of surgical or radiation intervention in connection with these tumors, brain injury as well as autoimmune, infiltrative and other rare diseases. Classical risk factors for functional CH include stress, excessive physical exercises and critically low body mass, however, variety of hormonal changes (including metabolic hormones) was observed and genetic predisposition was also shown for these patients. It is very important to understand complex pathogenetic factors that will help to improve diagnosis of this rare disease.

Effect of Air Pollution on Menstrual Cycle Length-A Prognostic Factor of Women's Reproductive Health.

Air pollution can influence women’s reproductive health, specifically menstrual cycle characteristics, oocyte quality, and risk of miscarriage. The aim of the study was to assess whether air pollution can affect the length of the overall menstrual cycle and the length of its phases (follicular and luteal). Municipal ecological monitoring data was used to assess the air pollution exposure during the monitored menstrual cycle of each of 133 woman of reproductive age. Principal component analyses were used to group pollutants (PM10, SO2, CO, and NOx) to represent a source-related mixture. PM10 and SO2 assessed separately negatively affected the length of the luteal phase after standardization (b = −0.02; p = 0.03; b = −0.06; p = 0.02, respectively). Representing a fossil fuel combustion emission, they were also associated with luteal phase shortening (b = −0.32; p = 0.02). These pollutants did not affect the follicular phase length and overall cycle length, neither in single- nor in multi-pollutant models. CO and NOx assessed either separately or together as a traffic emission were not associated with overall cycle length or the length of cycle phases. Luteal phase shortening, a possible manifestation of luteal phase deficiency, can result from fossil fuel combustion. This suggests that air pollution may contribute to fertility problems in women.

The role of vitamin D in reproductive dysfunction in women - a systematic review.

Vitamin D is essential for the proper functioning of the human body. There is also evidence of its strong association with fertility problems in women. This review aims to evaluate the relationship between vitamin D and diseases affecting women's fertility (polycystic ovarian syndrome (PCOS), uterine leiomyomas and endometriosis), and in vitro fertilization (IVF) outcome. A systematic review of the literature was conducted in Scopus and PubMed for relevant English language publications since 1989. Vitamin D influences the functioning of the reproductive system in women and has been associated with PCOS, uterine leiomyomas, endometriosis and in vitro fertilization (IVF) outcome. However, further studies on larger groups of patients are needed to establish what role vitamin D plays in the treatment of female infertility.

Transforming growth factor β signaling in uterine development and function.

Transforming growth factor β (TGFβ) superfamily is evolutionarily conserved and plays fundamental roles in cell growth and differentiation. Mounting evidence supports its important role in female reproduction and development. TGFBs1-3 are founding members of this growth factor family, however, the in vivo function of TGFβ signaling in the uterus remains poorly defined. By drawing on mouse and human studies as a main source, this review focuses on the recent progress on understanding TGFβ signaling in the uterus. The review also considers the involvement of dysregulated TGFβ signaling in pathological conditions that cause pregnancy loss and fertility problems in women.

Su1430 Risk of Congenital Anomaly in Children Born to Women With Coeliac Disease: A United Kingdom Population-Based Cohort Study

INTRODUCTION Several studies have raised the spectre of infertility being associated with coeliac disease. These have primarily been carried out in small groups of women attending infertility specialist services and subsequently screened for coeliac disease. Although nutritional deficits can contribute to infertility, such study designs are highly prone to produce biased estimates of the risk of fertility problems in women with coeliac disease in the general population. We carried out a large population-based study to compare clinical reporting of fertility problems in women with and without coeliac disease in the United Kingdom (UK). METHODS In the UK, any first contact or treatment from specialist infertility services requires referral from a women's primary care doctor, so this is the first clinical contact for assessment of fertility problems. Using The Health Improvement Network, a large primary care database of patients from across the UK, we estimated age-specific rates of new clinically recorded fertility problems in women with and without coeliac disease (CD) during their childbearing years between 1990 and 2013. These rates were then stratified by whether the fertility problem was recorded before or after the CD diagnosis and were compared to the general population rates using Poisson regression with adjustments for socioeconomic status and calendar time. RESULTS Out of the total population of 2,426,225 women, 6,506 (0.3%) women had a diagnosis of CD. The age specific-rates of new clinically recorded fertility problems in women with CD were very similar to the general population rates (11.6/1,000 person years in women with CD and 12.6/1,000 person years in the general population in the 30-34 year age group) (table 1). Incident fertility problems were recorded a median of 2 months (interquartile range: 4 years before, 2.7 years after). When stratified by the time of CD diagnosis the rates of clinically recorded fertility problems were very similar to the general population in both the time before and after CD diagnosis. However these rates were 50% higher after CD diagnosis in women aged 25-29 years compared to the same age group in the general population (IRR 1.50, 95% CI 1.10-2.04). Approximately 25% of the fertility problems were reported within a year before/after the CD diagnosis (Figure 1). CONCLUSION Women with coeliac disease do not have a substantially greater likelihood of reporting fertility problems than women without the condition. Rates were slightly higher in younger women with diagnosed CD, however as this effect did not hold for women of other age groups, increased ascertainment of the condition rather than a causal association may explain this. Table 1 Age specific rates of incident fertility problem reporting in women with and without coeliac disease

Su1429 Do Women With Coeliac Disease Clinically Present With Fertiliy Problems More Often Than Women in the General Population?

INTRODUCTION Several studies have raised the spectre of infertility being associated with coeliac disease. These have primarily been carried out in small groups of women attending infertility specialist services and subsequently screened for coeliac disease. Although nutritional deficits can contribute to infertility, such study designs are highly prone to produce biased estimates of the risk of fertility problems in women with coeliac disease in the general population. We carried out a large population-based study to compare clinical reporting of fertility problems in women with and without coeliac disease in the United Kingdom (UK). METHODS In the UK, any first contact or treatment from specialist infertility services requires referral from a women's primary care doctor, so this is the first clinical contact for assessment of fertility problems. Using The Health Improvement Network, a large primary care database of patients from across the UK, we estimated age-specific rates of new clinically recorded fertility problems in women with and without coeliac disease (CD) during their childbearing years between 1990 and 2013. These rates were then stratified by whether the fertility problem was recorded before or after the CD diagnosis and were compared to the general population rates using Poisson regression with adjustments for socioeconomic status and calendar time. RESULTS Out of the total population of 2,426,225 women, 6,506 (0.3%) women had a diagnosis of CD. The age specific-rates of new clinically recorded fertility problems in women with CD were very similar to the general population rates (11.6/1,000 person years in women with CD and 12.6/1,000 person years in the general population in the 30-34 year age group) (table 1). Incident fertility problems were recorded a median of 2 months (interquartile range: 4 years before, 2.7 years after). When stratified by the time of CD diagnosis the rates of clinically recorded fertility problems were very similar to the general population in both the time before and after CD diagnosis. However these rates were 50% higher after CD diagnosis in women aged 25-29 years compared to the same age group in the general population (IRR 1.50, 95% CI 1.10-2.04). Approximately 25% of the fertility problems were reported within a year before/after the CD diagnosis (Figure 1). CONCLUSION Women with coeliac disease do not have a substantially greater likelihood of reporting fertility problems than women without the condition. Rates were slightly higher in younger women with diagnosed CD, however as this effect did not hold for women of other age groups, increased ascertainment of the condition rather than a causal association may explain this. Table 1 Age specific rates of incident fertility problem reporting in women with and without coeliac disease

PHP147 - Astrological Signs and Health Problems; Demonstrating the Multiple Comparisons Problem

Astrological signs have been linked to numerous health problems, not only in pseudoscientific but also in high impact, peer-reviewed scientific literature. The aim of this study was to use a large prescription database to assess associations between astrological star signs and health problems. This study may demonstrate important statistical issues that arise when doing multiple comparisons. For this cross-sectional study, patient and drug data were collected from a large prescription database in The Netherlands. Patients were followed for one year, starting from their birthday in 2010. Outcomes were drug prescriptions for one of four health problems: cardiovascular disease, depression, erectile dysfunction in men and fertility problems in women. The “determinant” was patients’ astrological sign (based on Western-culture horoscope). Differences in prevalence rates between star signs were tested using logistic regression, adjusted for age and gender. Significance was set at P=0.05. Almost 300,000 persons were included for analysis. In our analysis, persons born under the star sign Aquarius were less likely to suffer from cardiovascular disease (P=0.001), whereas patients born under Scorpio or Sagittarius were more likely (P=0.011 and P=0.017). There were no effects on depression or erection dysfunctions. Women born under Gemini were 37% less likely to use fertility drugs (P=0.016). This study found several associations between star signs and health problems, none of which had any biologic plausibility. Interestingly, datasets filled with randomly generated star sign data also resulted in significant findings. Multiple comparisons without prespecified hypotheses, biological plausibility and adjustments for familywise error rates, can lead to false-positive findings. Multiple comparisons are common in outcomes research, appropriate use of statistics therefore is important.

Occurrence of fertility problems presenting to primary care: population-level estimates of clinical burden and socioeconomic inequalities across the UK

What are the age-specific incident rates of clinically recorded fertility problems in women aged 15-49 years and how do they vary by socioeconomic group and geographic area. The incident rate of recorded fertility problems was highest in women age 30-34 years: about 1% of women per annum. Overall rates did not vary by socioeconomic group; however, age-specific rates varied substantially by socioeconomic deprivation quintile; among younger women, deprivation was associated with higher infertility rates. The rates of infertility in the UK range from 2 to 26%. Infertility definitions and denominators vary widely, and most current evidence is based on questionnaire studies that are subject to recall, reporting and selection bias. The current paper presents population-based estimates of clinically recorded fertility problems in women of reproductive age and the variation by age and socioeconomic deprivation quintile across different regions of the UK, using a nationally representative cohort of women that is larger than any previous study. Although infertility overall does not vary by socioeconomic status, consultation for fertility problems is closely related to socioeconomic patterns of women's age at first conception, demonstrating that many couples have pre-existing, rather than specifically age-related, infertility. This cohort study used data from The Health Improvement Network, a computerized primary care database of anonymized patient records from general practices across the UK, with prospective health records on over 1.7 million women between 1990 and 2010. Our cohort included 1,776,746 women of reproductive age (age 15-49 years) who contributed one or more years of active general practice registration. We estimated rates of new clinically recorded fertility problems in these women using medical records and medications exclusively used to treat infertility. We assessed variation in age-specific incidence by socioeconomic deprivation quintile and geographic area using Poisson regression. The rate of incident recorded fertility problems was highest in women in the 30-34 year age group (10.9 per 1000 person-years), which equates to approximately 1% of women per annum in this age group. Lowest rates were in women in the 15-19 and 45-49 year age groups (0.7 and 0.4 per 1000 person-years, respectively). Overall rates did not vary by socioeconomic group, measured using quintiles of the Townsend index. Age-specific rates, however, varied substantially with socioeconomic deprivation quintile (P-value for interaction < 0.0001) such that up to age 25, women with more deprivation had more recorded fertility problems [rate ratio (RR) comparing most to least deprived 5.6, 95% confidence interval (CI) 4.4-7.2 at 15-20 years of age]. This reversed from age 25 to 39, when women with more deprivation had fewer recorded fertility problems (RR 0.6 95% CI 0.5-0.6 at age 30-34). After age 40, there was no socioeconomic gradient in absolute rates. This is by far the largest population-based study to estimate clinically recorded fertility problems in women and the first in the UK to assess variation across such a broad age group from 15 to 49 years. Our data, however, did not capture women who experience difficulty in conceiving, but do not consult their general practitioner (GP) regarding fertility problems. Compared with existing estimates, our measures of the extent and distribution of recorded fertility problems in primary care are more useful for GPs, primary care trusts and policy makers for the planning and delivery of fertility services. We have shown a high burden of infertility with little geographic variation; however, the significant burden in young, more deprived women needs recognition in light of age restrictions for treatment availability for infertility in the UK. Not only does treatment access need to be universal and more equitably allocated across socioeconomic groups, but also more resources are required to reduce fertility problems by targeting modifiable risk factors. There was no direct source of funding for this research work. N.N.D. completed the work as part of an M.Sc., which was funded by Developing Solutions Scholarship provided by the International Office, University of Nottingham. J.W. is supported by a University of Nottingham/National Institute for Health Research (NIHR) Senior Clinical Research Fellowship. Not applicable.

PW01-98 - No predictive value of anxiety- and depression symptoms for sub- or infertility

ObjectiveMental distress has been suggested as an etiological factor of fertility problems. The aim of the study was to investigate the predictive value of common mental symptoms for fertility problems in a population sample.MethodsThe predictive value of anxiety and depression symptoms for incident fertility problems over an 11 year span was investigated in a N=5,873 female sub-sample from the Nord-Trøndelag Health Studies (HUNT 1 and HUNT 2). Only women who had not experienced fertility problems at the time of HUNT 1 were included. Fertility problems were reported retrospectively at HUNT 2. Sub-/infertility was defined as having tried to get pregnant for more than one year without success. Symptoms of anxiety and depression at HUNT 1 were measured by the one-dimensional 12-item Anxiety Depression Index (ADI).ResultsMean age at HUNT 2 was 42 years (S.D. 4.96, range 28-49). N=152 reported having tried to get pregnant for more than one year. No predictive value of anxiety and/or depression symptoms for sub-/infertility was found (crude OR=0.97 (95% CI=0.82; 1.15), p=0.736; analyses adjusted for age, level of education, civil status, somatic conditions, parity, and gynaecological surgery: OR=0.97 (95% CI=0.81; 1.15), p=0.687). Results were comparable in a sub-sample who had never been pregnant.ConclusionThere is no prospective effect of common mental health symptoms on incident fertility problems in women. This zero-finding emerges from a large population-based data set with a long follow-up interval, and it provides evidence against the hypothesised causal relationship between mental distress and sub-/infertility.

119 VITRIFICATION OF IMMATURE FELINE OOCYTES: EFFECTS ON ZONA PELLUCIDA ULTRASTRUCTURE AND DEVELOPMENTAL COMPETENCE

Oocyte cryopreservation has been used to restore fertility problems in women undergoing oocyte-toxic chemotherapy and also to salvage the genetic potential of endangered/valuable species. In domestic cats, knowledge regarding the cryopreservation technique of feline oocytes is rather limited. We examined the effect of OPS vitrification on zona pellucida ultrastructure and also the developmental competence of immature cat oocytes. Immature cat oocytes were vitrified using different CPA and CPA exposure techniques: (1) 2-step DMSO, n = 71; (2) 4-step DMSO, n = 78; (3) 2-step EG, n = 73; (4) 4-step EG, n = 117; (5) 2-step EG + DMSO, n = 67 and (6) 4-step EG + DMSO, n = 88. A combination of EG and DMSO or EG alone using 4-step equilibration technique yielded the highest maturation rates (28/88: 31.8% and 44/117: 37.6%, respectively). This was, however, significantly lower than nonvitrified controls (36/59, 61% maturation rate). We further examined the effect of OPS vitrification on ultrastructure of the zona pellucida using scanning electron microscopy and also to test the quality of the zona pellucida using sperm binding assay. Our modified OPS protocols did not significantly affect the appearance of zona pellucida of vitrified oocytes and also the number of tightly bound sperm as compared with controls (80.8 ± 63.8 v. 81.6 ± 70.4, respectively). Vitrification of immature oocytes significantly reduced the number of oocytes reaching cleavage and blastocyst stages compared with nonvitrified controls (24.8 and 7.5% v. 62.5 and 30.8% for cleavage and blastocyst formation rates, respectively). It is concluded that modification of OPS vitrification can be successfully applied for immature cat oocytes, in terms of zona pellucida ultrastructure, ability to bind sperm, and also their developmental competence. This study was financially supported by MRG4980108, CHE-TRF Senior Research Scholars RTA-5080010, The Thailand Research Fund, and The Research Unit of Reproductive and Biological Science, Chulalongkorn University.