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Open access

Jennifer E Pearson-Farr, Gabrielle Wheway, Maaike S A Jongen, Patricia Goggin, Rohan M Lewis, Ying Cheong, and Jane K Cleal

Endometrial glands are essential for fertility, consisting of ciliated and secretory cells that facilitate a suitable uterine environment for embryo implantation. This study sought to determine whether an endometrial gland-specific transcriptome and splicing profile are altered in women with recurrent pregnancy loss. Our data provide a comprehensive catalogue of cilia and progestagen-associated endometrial protein (PAEP) gene isoforms and relative exon usage in endometrial glands. We report a previously unannotated endometrial gland cilia transcript GALNT11 and its susceptibility to exon skipping. Key endometrial receptivity gene transcripts are also reported to change in endometrial glands of women with recurrent pregnancy loss. The endometrial gland cilia and PAEP targets identified in this study could be used to identify a perturbed endometrium, isolate causes of recurrent pregnancy loss and develop targeted therapies in personalised medicine.

Lay summary

Successful embryo implantation is a trade-off between the lining of the womb which receives an implanting embryo, termed the endometrium, and a good quality embryo. For days 21–24 of the menstrual cycle, the endometrium undergoes changes into a receptive state in which it can receive an implanting embryo. Inappropriate endometrial receptivity is thought to underlie recurrent pregnancy loss. Improving pregnancy success in women with recurrent pregnancy loss requires an increased understanding of the endometrium at the molecular level. Genes contain the instructions for the cell and which genes are turned on or off determine how well it can do its role. We sought to determine a gene expression pattern of human endometrial glands in women with recurrent pregnancy loss (n = 5) vs a control group (n = 5). We identify target genes altered in women with recurrent pregnancy loss. Endometrial gland markers could be used to identify inappropriate endometrial receptivity.

Open access

Madara S B Ralapanawe, Sugandika Lakmali Gajaweera, Nishendra Karunaratne, Malcolm James Price, Pedro Melo, Arri Coomarasamy, and Ioannis Gallos

The use of intracytoplasmic sperm injection (ICSI) has recently increased worldwide. The live birth rate per ICSI cycle is low, and over half of infertile couples remain childless. Chromosomal polymorphisms are up to five times more common in couples with infertility compared to the general population. We aimed to investigate the association between chromosomal polymorphisms and reproductive outcomes in couples undergoing ICSI treatment. We analysed 942 ICSI fresh and frozen embryo transfer cycles in 697 women who underwent karyotyping analysis using Giemsa-Trypsin-Leishman banding prior to assisted conception at the Fertility Centre of Lanka Hospitals, Sri Lanka, between 2016 and 2018. The primary outcomes were pregnancy, miscarriage, and live birth rates. We compared outcomes according to the presence or absence of chromosomal polymorphism in females, males and couples. There were 294 pregnancies (31.2%) recorded in the study; 130 suffered a miscarriage (13.8%), 13 were ectopic pregnancies (1.3%) and 151 resulted in a live birth (16.0%). The evidence from univariable and multivariable analyses (adjusted for age, BMI, ovarian reserve and treatment type) did not confidently identify a difference in pregnancy, miscarriage or live birth rates between couples with no chromosomal polymorphisms compared to couples where the female, male or both partners were carriers of a chromosomal polymorphism. Further, we did not identify a clear association between the presence of chromosomal polymorphisms and reproductive outcomes compared to participants without chromosomal polymorphisms. Wide CIs precluded the identification of clinically meaningful associations.

Lay summary

Infertility affects approximately one in eight couples worldwide. The use of intracytoplasmic sperm injection (ICSI), where the sperm is directly injected into an egg using a micromanipulator outside the body, has become particularly popular in recent years. However, the success rate remains low. In human cells, the genetic material is arranged in structures called chromosomes. Chromosomal polymorphism is a normal variation where the genetic material is arranged differently to the average individual and is more common in infertile couples compared to the general population. We analysed data from 942 ICSI cycles in 697 couples who underwent karyotyping analysis to assess the changes in chromosomes between 2016 and 2018. The pregnancy rate was 31.2%, with 16.0% of participants experiencing a live birth, while 13.8% of pregnancies resulted in a miscarriage and 1.3% were outside the womb cavity (ectopic). The evidence did not identify a clear association between the chromosomal polymorphism and the outcome of treatment.

Open access

Perihan Sezginer, Cigdem Elmas, and Fatma Yıldız

Graphical abstract

Abstract

This study was carried out to investigate whether repeated controlled ovarian hyperstimulation (COH) affects ovarian reserve. For this reason, we aimed to show possible changes in the expression of PTEN and FOXO3, which are involved in preserving the over-reserve, after applying the COH protocol methods. For this purpose, 18 young Wistar albino female rats (8 weeks old) were randomly assigned as group 1 (control), group 2, and group 3 as 6 subjects in each group. Experimental groups were treated with 10 IU/0.1 mL pregnant mare’s serum gonadotropin and a COH protocol consisting of 10 IU/0.1 mL human chorionic gonadotropin injection after 48 h. This procedure was applied three and five times to group 2 and group 3, respectively. For the control groups, the same procedures were performed with 0.1 mL of 0.9% sodium chloride solution. At the end of the experiment, the ovarium tissues were placed in a 10% neutral formaldehyde solution for light microscopic examinations. In histological sections stained with hematoxylin and eosin, the number of ovarian follicles was determined using the physical dissector method. However, the expression of PTEN, FOXO3, and LH-R molecules was evaluated by immunohistochemical methods. As a result of our study, it was concluded that COH administration reduces the expression levels of PTEN and FOXO3 proteins and LH-R, which are among the essential components of the PIK3 intracellular signaling pathway and also increased the levels of hormones such as follicle-stimulating hormone, estradiol, and luteinizing hormone, which are over-reserve markers, and causes adverse effects on the histological structure, oocyte morphology, and number of ovaries.

Lay summary

Today, approximately 10–15% of couples experience fertility problems. However, assisted reproductive techniques help people with fertility problems to get pregnant. The main purpose of these techniques is to put the sperm and egg together outside the woman’s body where the eggs are fertilized and then to return the fertilized eggs (embryos) to the womb. During a woman’s menstrual cycle, several hormones influence the growth of the eggs. This process can be mimicked by using various medications. Medication is given to increase the number of eggs that develop. However, this method is not the same as normal ovulation. Therefore, in our study, we wanted to examine the effect that developing multiple follicles has on the number and quality of eggs remaining for the future.

Open access

Andrea Romano, Sun-Wei Guo, Jan Brosens, Asgerally Fazlebas, Caroline E Gargett, Stefan Giselbrecht, Martin Gotte, Linda Griffith, Hugh S Taylor, Robert N Taylor, Hugo Vankelecom, Charles Chapron, Xiao-Hong Chang, Khaleque N Khan, and Paola Vigano’

In vitro

culturing of endometrial cells obtained from the uterine mucosa or ectopic sites is used to study molecular and cellular signalling relevant to physiologic and pathologic reproductive conditions. However, the lack of consensus on standard operating procedures for deriving, characterising and maintaining primary cells in two- or three-dimensional cultures from eutopic or ectopic endometrium may be hindering progress in this area of research. Guidance for unbiased in vitro research methodologies in the field of reproductive science remains essential to increase confidence in the reliability of in vitro models. We present herein the protocol for a Delphi process to develop a consensus on in vitro methodologies using endometrial cells (ENDOCELL-Seud Project). A steering committee composed of leading scientists will select critical methodologies, topics and items that need to be harmonised and that will be included in a survey. An enlarged panel of experts (ENDOCELL-Seud Working Group) will be invited to participate in the survey and provide their ratings to the items to be harmonised. According to Delphi, an iterative investigation method will be adopted. Recommended measures will be finalised by the steering committee. The study received full ethical approval from the Ethical Committee of the Maastricht University (ref. FHML-REC/2021/103). The study findings will be available in both peer-reviewed articles and will also be disseminated to appropriate audiences at relevant conferences.

Lay summary

Patient-derived cells cultured in the lab are simple and cost-effective methods used to study biological and dysfunctional or disease processes. These tools are frequently used in the field of reproductive medicine. However, the lack of clear recommendations and standardised methodology to guide the laboratory work of researchers can produce results that are not always reproducible and sometimes are incorrect. To remedy this situation, we define here a method to ascertain if researchers who routinely culture cells in the lab agree or disagree on the optimal laboratory techniques. This method will be used to make recommendations for future researchers working in the field of reproductive biology to reproducibly culture endometrial cells in the laboratory.

Open access

Ezekiel O Mecha, Joseph N Njagi, Roselydiah N Makunja, Charles O A Omwandho, Philippa T K Saunders, and Andrew W Horne

Endometriosis has long been wrongly perceived to be rare among women of African descent. The misconception about the prevalence of endometriosis among African women has significantly contributed to long diagnostic delays, limited access to diagnosis and care, and a scarcity of research on the condition among African women. In this commentary, we highlight the prevalence of endometriosis among African women, the state of endometriosis care in Africa, and the gaps in knowledge that need to be addressed. Based on the available data, the prevalence of endometriosis in Africa is likely higher than previously thought, with varying subtypes. There is a long diagnostic delay of endometriosis among African women. Additionally, endometriosis care in Africa from the general population and health practitioners is poor; this can be attributed to the high diagnostic cost, scarcity of trained specialists, as well as patients’ inability to express their symptoms due to societal taboos surrounding menstrual health. Public sensitization on endometriosis may help improve endometriosis diagnosis and care in Africa.

Lay summary

Endometriosis is a condition in which tissue like the uterine lining is found outside the uterus, causing women to experience pain especially before, during, or after menstruation. Although endometriosis affects an estimated 176 million women worldwide, it has been wrongly reported that endometriosis is a rare condition among African women, mainly due to lack of awareness among healthcare providers and historical bias. In the current commentary, we discuss the prevalence of endometriosis, the diagnostic delays, and the care of endometriosis among black African women living in the African continent. Much of the literature has demonstrated (falsely) that endometriosis is rare in Black women compared to White ethnicity. African women experience a long diagnostic delay and do not receive appropriate care. Public awareness of endometriosis may help improve diagnosis delay and endometriosis care in Africa.

Open access

Emmanuel Amabebe, Noopur Bhatnagar, Nitin Kamble, Steven Reynolds, and Dilly O Anumba

Lay summary

The need to develop new treatments to prevent unprompted premature delivery before 37 weeks of pregnancy remains pressing and unmet. Bacteria (Lactobacillus species) that promote vaginal health produce biochemical compounds that prevent the growth of microbes such as Gardnerella vaginalis. Overgrowth of G. vaginalis can cause vaginal infection with smelly discharge and increase a woman’s risk of sexually transmitted infections and premature delivery. In this study, we examined how normal health-promoting (L. crispatus) and potentially harmful (G. vaginalis) vaginal bacteria interact in a laboratory setting. This was in order to observe natural and effective agent(s) from L. crispatus that can hinder the growth of G. vaginalis and accompanying immune response. We observed that L. crispatus clears G. vaginalis by itself and with several biochemical compounds that it produces. Such biochemical compounds can be developed into treatment for vaginal infections and premature delivery due to infection and inappropriate immune response.

Open access

Gisele Montano, Pat Clough, Todd Schmitt, Michelle Davis, Karen Steinman, Justine O’Brien, and Todd Robeck

To gain more knowledge about the influence of hormone regulation on follicle development, ovarian ultrasounds were performed, and urinary hormone profiles were determined in ovulating and non-ovulating female bottlenose dolphins (n  = 15) following estrus synchronization with altrenogest. Ovarian ultrasounds were conducted daily, post-synchronization to describe follicular recruitment in relation to the endocrine profile. Follicle sizes were grouped into very small (VSM), small (SM), medium (MD) and large (LG). In ovulating females, two follicular waves were identified, and follicular deviation toward establishing a dominant follicle only occurred during the second wave. For non-ovulating females, only the first wave was observed. For all urinary hormones, the non-ovulating group presented significantly lower concentrations of follicle-stimulating hormone (uFSH), luteinizing hormone (uLH), estrone conjugates (uE1-C) and estriol (uE3) but similar progestagen and cortisol concentrations compared to the ovulating group. Concentrations of uE1-C and uE3 and the numbers of MD and LG follicles significantly (P < 0.05) increased, while uFSH concentrations significantly (P < 0.05) decreased as ovulation approached. Urinary LH significantly increased concurrently with increasing numbers of LG follicles and decreasing numbers of SM follicles. The characterization of follicular development and its relationship with hormone assessment complements our understanding of follicular recruitment post-synchronization in bottlenose dolphins and provides new information concerning differences between ovulating and non-ovulating females in response to an estrous synchronization protocol.

Lay summary

Many species of fresh and saltwater dolphins and porpoises are facing increasing pressure for food and habitats due to human activities. One of the primary aspects that can be affected by such activities is reproduction. The bottlenose dolphin has been successfully bred in zoos and aquariums for at least 70 years, and they can be trained for several voluntary behaviors that enable medical examinations. Therefore, they represent a unique resource for research and understanding of normal dolphin reproduction. In this research, voluntary ultrasound exams and urine samples from 15 female dolphins were used to describe changes in their ovaries. The resulting descriptions and comparisons provide insight into the unique ovarian physiology of bottlenose dolphins and into the differences between animals after treatment. This information adds to the body of knowledge which one day may be used for developing advanced reproductive techniques to help preserve endangered species of dolphins.

Open access

Nicola Tempest, Madeleine France-Ratcliffe, Hannan Al-Lamee, Evie R Oliver, Emily E Slaine, Andrew J Drakeley, Victoria S Sprung, and Dharani K Hapangama

Optimisation of lifestyle factors such as smoking and alcohol is encouraged to improve fecundability rates in the fertility setting. Currently, routine fertility consultations do not involve counselling or imparting advice regarding habitual physical activity (PA) and/or structured exercise, despite data showing that vigorous PA can be associated with delayed time to pregnancy. Therefore, this study aimed to determine habitual PA in a sample of women attending the one-stop infertility (OSI) clinic. In total, 250 women attending a large tertiary level NHS fertility unit prospectively completed a questionnaire anonymously over a period of 9 months. The participants (mean age: 34 ± 5 years, mean BMI: 29 ± 7 kg/m2) habitual PA levels varied from vigorous exercise on ≥5 days/week (8%, n  = 17) to no moderate or high-intensity activities across the whole week (66%, n  = 29). A majority of women reported no structured exercise (72%, n  = 179). No association was identified between any domain of PA and BMI, age, alcohol units, regular periods, or time spent trying to conceive (P > 0.05). The participants habitual PA levels varied widely and no association between any domain of PA and background of the women was identified. No existing evidence and/or guidelines to explicitly inform women attempting to conceive regarding recommended PA levels are available, despite PA being a modifiable, affordable, and feasible lifestyle choice with the possible potential to improve fertility. A large-scale, clinical trial assessing the effects of PA on fecundability is warranted to gain insight into the potential of this lifestyle factor to improve fertility outcomes and to explore the underlying biological mechanisms involved.

Lay summary

Infertility affects approximately one in seven couples with many and varied causes, including lifestyle factors such as smoking, alcohol, and diet. Lifestyle changes are low-cost unimposing options to implement in routine fertility care. Information on regular physical activity is not currently enquired from women and no agreement regarding the best levels of exercise exists for fertility patients. In this study, we aimed to determine the exercise habits of women attending the OSI clinic. In total. 250 women attending OSI clinic over a period of 9 months completed a questionnaire collecting data on their exercise habits. The levels of physical activity performed varied widely from vigorous exercise on ≥5 days/week to no moderate or high-intensity activities across the whole week. A majority of women did no structured exercise (72%). These novel data highlight the variations in form, type and intensity of exercise women who attend OSI clinics perform. Currently, there is no existing evidence and/or guidelines to explicitly inform women attempting to conceive regarding the recommended physical activity levels. Physical activity is a modifiable, affordable, and feasible lifestyle choice that is not currently acknowledged in the fertility setting and has the potential to improve fertility.

Open access

Mike Armour, Jodie Avery, Mathew Leonardi, Leesa Van Niekerk, Marilla L Druitt, Melissa A Parker, Jane E Girling, Brett McKinnon, Antonina Mikocka-Walus, Cecilia H M Ng, Rebecca O’Hara, Donna Ciccia, Katherine Stanley, and Subhadra Evans

Graphical abstract

Abstract

Endometriosis is a common yet under-recognised chronic disease with one in nine (more than 830,000) women and those assigned female at birth diagnosed with endometriosis by the age of 44 years in Australia. In 2018, Australia was the first country to develop a roadmap and blueprint to tackle endometriosis in a nationwide, coordinated manner. This blueprint is outlined in the National Action Plan for Endometriosis (NAPE), created from a partnership between government, endometriosis experts and advocacy groups. The NAPE aims to improve patient outcomes in the areas of awareness and education, clinical management and care and research. As researchers and clinicians are working to improve the lives of those with endometriosis, we discuss our experiences since the launch of the plan to highlight areas of consideration by other countries when developing research priorities and clinical plans. Historically, major barriers for those with endometriosis have been twofold; first, obtaining a diagnosis and secondly, effective symptom management post-diagnosis. In recent years, there have been calls to move away from the historically accepted ‘gold-standard’ surgical diagnosis and single-provider specialist care. As there are currently no reliable biomarkers for endometriosis diagnosis, specialist endometriosis scans and MRI incorporating artificial intelligence offer a novel method of visualisation and promising affordable non-invasive diagnostic tool incorporating well-established technologies. The recognised challenges of ongoing pain and symptom management, a holistic interdisciplinary care approach and access to a chronic disease management plan, could lead to improved patient outcomes while reducing healthcare costs.

Lay summary

Endometriosis is a chronic disease where tissue like the lining of the uterus is found in other locations around the body. For the 830,000 people living with endometriosis in Australia, this often results in an immense burden on all aspects of daily life. In 2018, Australia was the first country to introduce a roadmap and blueprint to tackle endometriosis in a nationwide coordinated manner with the National Action Plan for Endometriosis. This plan was created as a partnership between government, endometriosis experts and advocacy groups. There are several other countries who are now considering similar plans to address the burden of endometriosis. As researchers and clinicians are working to improve the lives of those with endometriosis, we share our experiences and discuss areas that should be considered when developing these national plans, including diagnostic pathways without the need for surgery, and building new centres of expertise in Endometriosis and Pelvic Pain.

Open access

Vanessa Caroline Fioravante, Alana Rezende Godoi, Victória Mokarzel de Barros Camargo, Patricia Fernanda Felipe Pinheiro, Marcelo Martinez, Carlos Roberto Padovani, and Francisco Eduardo Martinez

Graphical abstract

Abstract

The relationship between adolescent ethanol uses and its impacts throughout life are not conclusive. Thus, we evaluated if the low and high consumption of ethanol at postpuberty interferes with the reproduction and ethanol-naive offspring and if the effects are dose-related. Female and male rats were divided into three groups: low drinker (L), high drinker (H) and control (C). The L and H groups were exposed to ethanol up to 10 % from 65 to 80 days with withdrawal after this period. The ethanol consumed by low drinkers was 1.41 ± 0.21 g/kg/day and by high drinkers 4.59 ± 0.45 g/kg/day. The study was conducted in two phases. The first phase verified the reproductive capacity in adulthood on generations (litter size and sex ratio). Data were collected over 10 years. The second phase analyzed the parent reproductive parameters (body weight, reproductive organ weight, sperm parameters and estrous cycle) and the pup development. We observed a reduced litter size in both drinker groups. Gestational body weight gain and feed consumption were lower in L and H. We observed an alteration in reproductive organs weight in both sexes of H. Females presented a longer estrous cycle duration. Males presented an increase in abnormal sperm, a decrease in sperm count and accelerated transit time. The ethanol-naive offspring development was also impaired. We conclude that low and high postpubertal alcohol use impairs long-term reproductive parameters, even after alcohol withdrawal. There is also impaired ethanol-naive offspring. Besides, the effects are dose-related.

Lay summary

The effects of alcohol use have been reported in several studies. However, better knowledge about early alcohol use and its impact on reproduction in adulthood, after abstinence and on ethanol-naive offspring could help improve preventive measures and mechanisms of action. One of the methods used was retrospective analysis which allows to evaluate the effects of postpubertal ethanol use on the reproductive capacity of rats over generations. Despite our limitations, we verified that the post-adolescent period acts as a susceptibility window, and lifestyle at this age modulates the long-term reproductive parameters. The early ethanol use impairs reproduction function since sperm parameters and the estrous cycle have been altered. The dose of alcohol also contributes to damage on the drinkers’ reproduction and on the physical development of ethanol-naive offspring. Future studies are necessary to identify the mechanism involved in long-term alcohol use effects, even in withdrawal, as well as ethanol-naive offspring outcomes.