Follicle-stimulating hormone receptor gene polymorphism in chronic anovulatory women, with or without polycystic ovary syndrome: a cross-sectional study

BACKGROUND: Polymorphisms at codons 307 and 680 are the most commonly encountered allelic variants of the follicle-stimulating hormone receptor (FSHR) gene. Studies in Caucasians suggest that certain FSHR variants are more common in women with polycystic ovary syndrome (PCOS) than normal women. The objective of this study was to determine the distribution of FSHR gene polymorphisms at codons 307 and 680 in Thai women with chronic anovulation, without (121 women) and with PCOS (133 women), using 132 known fertile women as controls. METHODS: DNA samples from peripheral blood lymphocytes were extracted and analyzed by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The prevalence of Threonine307Threonine (TT), Threonine307Alanine (TA), and Alanine307Alanine (AA) genotypes at codon 307 was 53.0% (95% CI = 44.2-61.7%), 42.4% (95% CI = 34–51.3%), and 4.5% (95% CI = 1.9-10.1%) in controls; 52.6% (95% CI = 43.8-61.3%), 39.8% (95% CI = 31.6-48.7%), and 7.5% (95% CI = 3.9-13.7%) in PCOS women; and 50.4% (95% CI = 42.8-61.2%), 45.4% (95% CI = 34.9-53.1%), and 4.5% (95% CI = 1.5-9.6%) in anovulatory women without PCOS, respectively. The prevalence of Asparagine680Asparagine (NN), Asparagine680Serine (NS), and Serine680Serine (SS) genotypes at codon 680 was 54.5% (95% CI = 45.7-63.2%), 40.9% (95% CI = 32.5-49.8%), and 4.5% (95% CI = 1.9-10.1%) in controls; 51.9% (95% CI = 43.1-60.6%), 44.4% (95% CI = 35.8-53.2%), and 3.8% (95% CI = 1.4-9.0%) in PCOS women; and 47.9% (95% CI = 40.4-58.8%), 47.1% (95% CI = 36.5-54.7%), and 5.0% (95% CI = 2–10.9%) in anovulatory women without PCOS, respectively. The prevalence of FSHR gene polymorphisms at both codons were not statistically different among the three groups. CONCLUSIONS: In Thai women, there was no association between the FSHR gene polymorphism at codons 307 and 680 and chronic anovulation

Ovarian follicular fluid reflects the clinical condition and oocyte cumulus homeostasis.

Infertility is a worldwide problem that is often overlooked. Although assisted reproductive technology has been developed over decades, many women still suffer from infertility. More knowledge is needed to understand ovarian homeostasis to optimise fertility treatment. This study aimed to explore the relationship of lipids and glucose levels in blood and follicular fluid, and compare these substrates among women with normal or abnormal metabolic condition. It also sought to measure lipid content within human oocytes as well as the expression of endoplasmic reticulum stress marker genes in cumulus cells, and their relationship with metabolic substances, obesity and IVF outcome. The blood, follicular fluid, cumulus cells and unfertilised oocytes from 88 women, who underwent IVF in FertilitySA from February 2011 to August 2011, were collected and analysed for glucose, lipids and endoplasmic reticulum stress markers. Follicular glucose, insulin, high density lipoprotein cholesterol (HDL-C) and majority of polyunsaturated fatty acid (PUFA) and monounsaturated fatty acid (MUFA) levels correlated with the serum levels (r= 0.16-0.23). Insulin was associated with the BMI, waist circumference, metabolic syndrome and many fatty acids, but not follicular glucose. However, the immaturity rate of the retrieved oocytes correlated with the follicular glucose and total fatty acids (r = 0.19-0.26, p <0.04). Variability of the unfertilised oocyte morphology correlated with follicular glucose, and the immaturity rate also differed among the metabolic syndrome group. An increase of follicular 18:3 n-3 (alpha-linolenic) and decrease of 20:3 n-3 (eicosatrienoic acid; ETA) existed in women with a waist circumference of more than 80 cm. The follicular 20:5 n-3 (eicosapentaenoic acid; EPA) percentage correlated with fertilisation and cleavage rate (r = -0.32, p = 0.003 and r = -0.35, p = 0.001). The follicular low density lipoprotein cholesterol (LDL-C) and HDL-C related to follicular fatty acids. Higher levels of serum LDL-C (2.31 ± 0.65 and 1.98 ± 0.61mmol/L, p 0.02) and follicular fatty acid (0.26 ± 0.09 and 0.22 ± 0.05 mmol/L, p = 0.03) were found in non-pregnant women. There was a wide variability of ER stress expression in cumulus cells among women in this study. There was no obvious correlation between ER stress markers and other measurements. The unfertilised oocyte BODIPY fluorescence intensity had high variability among women and individuals. However, the unfertilised oocyte lipid content correlated with the serum LDL-C level. Substances with a good follicular-serum relationship may be transported directly from blood to the follicle. The discorrelation might be affected by intrafollicular metabolism. Insulin may be involved in follicular lipid metabolism because it correlated with many follicular fatty acids and cholesterols. The equilibrium between follicular fatty acids involving insulin modulation may affect oocyte quality. Overall, this study found there were correlations between serum and follicular lipids, follicular insulin and cholesterol with follicular fatty acids and the importance of serum LDL-C and follicular omega-3 fatty acids. Serum insulin and LDL-C screening might be another tool for predicting the follicular lipid dysequilibrium and poor IVF outcome. The unfertilised oocyte may be a useful tool for a study on oocyte quality. A larger study is needed to recruit more women of different ages and BMI for a stronger correlation between follicular insulin, glucose and lipid metabolism, and ER stress markers.Thesis (M.Med.Sc.) -- University of Adelaide, School of Paediatrics and Reproductive Health, 201

The effect of different progestogens on sleep in postmenopausal women: a randomized trial

<p><b>Background:</b> While progesterone affects sleep, different types of it might affect sleep differently.</p> <p><b>Methods:</b> One hundred Thai women, who complained of insomnia, visited the Menopause Clinic at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand from February 2014 to March 2015, and were divided randomly into two groups. Both groups received daily hormonal treatment that included estradiol valerate (progynova) at 1 mg. The first group also received dydrogesterone (duphaston^®) at 10 mg and the second group micronized progesterone (utrogestran^®) at 100 mg. The clinical symptoms and Pittsburgh Sleep Quality Index (PSQI) were recorded for three consecutive months after treatment. This study was registered with clinicaltrial.gov (code number NCT02086032).</p> <p><b>Results:</b> Sleep quality improved in both groups (10.52 ± 4.27 to 4.91 ± 3.15 in the dydrogesterone group and 10.16 ± 3.60 to 6.27 ± 3.04 in the micronized progesterone group, <i>p</i> value 0.08). Women in the micronized progesterone group had fewer overall side effects than those in the dydrogesterone group.</p> <p><b>Conclusion:</b> Sleep quality of peri-postmenopausal women with insomnia improved dramatically after the first month of hormonal treatment. However, more participating patients are necessary to ascertain the differences in sleep quality from dydrogesterone and micronized progesterone treatment.</p

Pre-implantation genetic testing for Marfan syndrome using mini-sequencing

Marfan syndrome (MFS1) is an autosomal dominant condition causing aortopathy including fatal aortic dissection. This study aimed to perform clinical PGT-M in a family with a history of MFS1 for two generations. A family with two members affected by MFS1 approached the hospital for PGT-M. The couple decided to join the project following extensive counselling and informed consent was obtained. The mutation contributory to MFS1 was identified using whole-exome sequencing (WES). A novel PGT-M protocol using multiplex fluorescent PCR and mini-sequencing was developed and tested. Ten blastocysts were subjected to PGT-M in two clinical PGT cycles. Mini-sequencing revealed four normal and six affected embryos. Microsatellite-based linkage analysis confirmed mutation analysis results in all samples. The embryos diagnosed as normal (non-MFS1) were chosen for transfer. A pregnancy was obtained in the third embryo transfer. Invasive prenatal diagnosis confirmed the normal genotype of the baby. This study demonstrated comprehensive management using the application of clinical-based diagnosis, WES for mutation identification within the MFS1 gene, mini-sequencing for embryo selection and microsatellite-based linkage analysis for backup of PGT-M results and contamination detection to assist couples in having a healthy child when there was a family history of Marfan syndrome.Impact Statement What is already known on this subject? Marfan syndrome (MFS1, OMIM#154700) is an autosomal dominant condition causing aortopathy including fatal aortic dissection. Pre-implantation genetic testing (PGT) is an alternative to traditional invasive prenatal diagnosis (PND) giving the couples the chance of starting pregnancy with the confidence that the baby will be unaffected. Most of the previous PGT reports employed microsatellite-based linkage analysis. A few PGT studies used sequencing, mini-sequencing and mutation analysis; however, the details of the techniques were not described. What do the results of this study add? Single-cell PCR protocol using multiplex fluorescent PCR and mini-sequencing was developed and validated. Two clinical PGTs cycles for Marfan syndrome were performed. A healthy baby was resulted. The details of multiplex fluorescent PCR and mini-sequencing protocols are described in this study so that the procedures can be reproduced. What are the implications of these findings for clinical practice and/or further research? Embryo selection can help the family suffering from Marfan syndrome for two generations to start a pregnancy with confidence that their child will be unaffected. This study also shows the use of a mini-sequencing protocol for PGT, which can be a universal protocol for other mutations by changing the PCR primers and mini-sequencing primers

The successful strategy of comprehensive pre-implantation genetic testing for beta-thalassaemia–haemoglobin E disease and chromosome balance using karyomapping

Thalassaemia is the commonest monogenic disease and causes a health and economic burden worldwide. Karyomapping can be used for pre-implantation genetic testing of monogenic disorders (PGT-M). This study applied karyomapping in two PGT-M cycles and made a comparison to polymerase chain reaction (PCR). Two families at risk of having beta-thalassaemia–haemoglobin E disease offspring decided to join the project and informed consent was obtained. Karyomapping results of family A (beta-thalassaemia (c.41_42delTCTT)-Hb E (c.26G>A) disease) revealed four normal, two beta-thalassaemia traits, one Hb E trait and six affected. Three embryos exhibited unbalanced chromosomes. One normal male embryo was transferred. Karyomapping results of family B (beta-thalassaemia (c.17A>T)-Hb E (c.26G>A) disease) revealed six Hb E traits and three affected. Three embryos were chromosomally unbalanced. One Hb E trait embryo was transferred. Two successful karyomapping PGT-M were performed, including deletion and single-base mutations. Karyomapping provides accuracy as regards the protocol and copy number variation which is common in pre-implantation embryos. Impact Statement What is already known on this subject? Thalassaemia syndrome is the commonest monogenic disease and causes a health and economic burden worldwide. Modern haplotyping using SNP array (aSNP) and karyomapping algorithms can be used for pre-implantation genetic testing of monogenic disorders (PGT-M). However, few clinical karyomapping PGT-M cycles have been done and validated so far. What do the results of this study add? Two successful clinical PGT-M cycles for beta-thalassaemia (c.41_42delTCTT and c.17A>T mutations)–haemoglobin E (c.26G>A) disease were performed using karyomapping. The outcome was two healthy babies. Multiplex fluorescent polymerase chain reaction (PCR) with mini-sequencing was also used for confirmation mutation analysis results. PCR confirmed haplotyping results in all embryos. Six embryos from both PGT-M cycles exhibited unbalanced chromosomes evidenced by aSNP. What are the implications of these findings for clinical practice and/or further research? Karyomapping provides accurate information quickly and the outcomes of the study will save time as regards protocol development, provide a usable universal PGT-M protocol and add additional copy number variation (CNV) information, chromosome number variation being a common issue in pre-implantation embryos

Thai Interest Group for Endometriosis (TIGE) consensus statement on endometriosis-associated pain

This consensus statement has been developed by the Thai Interest Group for Endometriosis (TIGE) for use by Thai clinicians in the diagnosis and management of endometriosis. TIGE is a group of clinical and academic gynaecologists with a particular interest in endometriosis. Endometriosis is an oestrogen-dependent inflammatory disease which causes chronic symptoms such as dysmenorrhoea, chronic pelvic pain, dyspareunia and subfertility, and it is common in reproductive-age women. There is limited overall data on its prevalence in different clinical settings in Thailand, but it is clear that the disease causes significant problems for patients in terms of their working lives, fertility, and quality of life, as well as placing a great burden on national healthcare resources. Decisions about selecting the appropriate treatment for women with endometriosis depend on many factors including the age of the patient, the extent and severity of disease, concomitant conditions, economic status, patient preference, access to medication, and fertility need. Several hormonal treatments are available but no consensus has been reached about the best option for long-term prevention of recurrence. Bearing in mind differences in environment, genetics, and access to the healthcare system, this treatment guideline has been tailored to the particular circumstances of Thai women

The effects of being overweight and obese on female reproduction: a review

Obesity is a major international problem related to many reproductive health problems including polycystic ovary syndrome (PCOS). This article reviews the evidence of being overweight and its effect on female reproduction. The fecundity of obese women is lower than normal weight women, but there is no absolute consensus about the effect of obesity on infertility treatment. The obese patient might have oocyte, hormone, metabolic and endometrial dysfunction affecting reproduction. Insulin and leptin may be some of the answers explaining anovulation during obesity leading to infertility. Moreover, the follicular glucose and lipids which are important for oocyte development also increase in the obese patient and these might have an effect on oocyte quality because studies in mice have revealed that the obesity affects follicular cell stress and oocyte lipids. Overall, obesity affects female reproduction by disturbing the general body metabolism, hormone metabolism and the follicular environment.Tawiwan Pantasri and Robert John Norma