Reproductive BioMedicine Online

2012-02-01

The Edwards and Steptoe Research Trust: commemorating the two great pioneers of our field and supporting their aspirations

Jacques Cohen, Gedis Grudzinskas, Martin H. Johnson — 2011-12-06

Trustees: Professor Martin Johnson FRCOG (Chief Executive Officer), Professor Peter Braude FRCOG, FMedSci, Ruth Edwards, Professor Sir Richard Gardner FRS, David Martin FCA (Treasurer) and Professor Andrew Steptoe FMedSci.

Useful lessons from China?

Jacques Cohen, Gedis Grudzinskas, Martin Johnson — 2012-02-01

Whilst Europe struggles with ethical, legal and commercial aspects of providing for the growing number of ageing women who seek fertility treatment using oocytes other than their own, our Chinese colleagues have developed a novel strategy within their own ethical, social and legal boundaries. In this issue, describe a clinical regime in which all patients are potential donors: surplus oocytes are vitrified at the time of oocyte collection and the question of their donation considered only if a satisfactory outcome is obtained. This approach must be considered a positive development.

Professor Edwin Carlyle (Carl) Wood AC, CBE, FRCS, FRCOG, FANZCOG

Alan Trounson — 2011-11-28

Carl Wood died on 23 September 2011 in Melbourne, Australia after a long battle with Alzheimer’s disease. He was 82years old. Carl was the foundation Chair of Obstetrics and Gynaecology, Monash University, Queen Victoria Hospital, Melbourne. He was a distinguished clinical scholar and researcher in the disciplines of both obstetrics and gynaecology. He was an outstanding surgeon with an innovative approach that set him aside from most academic leaders in reproductive medicine. He was always willing to try the most creative methods and collaborated with others to enable the rapid evolution of new science. He was recognized by elite medical practitioners and researchers in reproductive medicine as a leader likely to change the world’s approach to patient care. Indeed he did.

The luteal phase after GnRH-agonist triggering of ovulation: present and future perspectives

2011-11-10

Abstract: In stimulated IVF/intracytoplasmic sperm injection cycles, the luteal phase is disrupted, necessitating luteal-phase supplementation. The most plausible reason behind this is the ovarian multifollicular development obtained after ovarian stimulation, resulting in supraphysiological steroid concentrations and consecutive inhibition of LH secretion by the pituitary via negative feedback at the level of the hypothalamic–pituitary axis. With the introduction of the gonadotrophin-releasing hormone-(GnRH) antagonist, an alternative to human chorionic gonadotrophin triggering of final oocyte maturation is the use of GnRH agonist (GnRHa) which reduces or even prevents ovarian hyperstimulation syndrome (OHSS). Interestingly, the current regimens of luteal support after HCG triggering are not sufficient to secure the early implanting embryo after GnRHa triggering. This review discusses the luteal-phase insufficiency seen after GnRHa triggering and the various trials that have been performed to assess the most optimal luteal support in relation to GnRHa triggering. Although more research is needed, GnRHa triggering is now an alternative to HCG triggering, combining a significant reduction in OHSS with high ongoing pregnancy rates.Stimulation of the ovaries for IVF treatment induces the growth of multiple follicles, resulting in the aspiration of several oocytes. During the early years of IVF treatment it became obvious that the stimulation per se induced an unphysiological hormonal level during the luteal phase – the phase after egg transfer – necessitating hormonal support with vaginally applied progesterone to obtain ongoing pregnancies. With the introduction of the gonadotrophin-releasing hormone (GnRH) antagonist protocol (short protocol) it became possible to perform final oocyte maturation with a GnRH agonist instead of human chorionic gonadotrophin (HCG). The first studies applying this concept, however, showed a very poor pregnancy rate, despite standard luteal-phase support with progesterone. This review discusses the reason for the poor results and the newest studies, using GnRH agonist instead of HCG, now showing a normalization of pregnancy rates due to modifications of the luteal-phase support and with the benefit of a protocol which has a reduced risk of ovarian hyperstimulation syndrome.

Microdrop preparation factors influence culture-media osmolality, which can impair mouse embryo preimplantation development

J.E. Swain, L. Cabrera, X. Xu, G.D. Smith — 2011-10-24

Abstract: Because media osmolality can impact embryo development, the effect of conditions during microdrop preparation on osmolality was examined. Various sizes of microdrops were prepared under different laboratory conditions. Drops were pipetted directly onto a dish and covered by oil (standard method) or pipetted on the dish, overlaid with oil before removing the underlying media and replaced with fresh media (wash-drop method). Drops were made at 23°C or on a heated stage (37°C) and with or without airflow. Osmolality was assessed at 5min and 24h. The biological impact of osmolality change was demonstrated by culturing 1-cell mouse embryos in media with varying osmolality. Reduced drop volume, increased temperature and standard method were associated with a significant increase in osmolality at both 5min and 24h (P-values <0.001, <0.0001 and <0.0001, respectively). There was a significant interaction between airflow, decreased volume, increased temperature and standard method that caused a significant increase in osmolality (40mOsm/kg) compared with controls (P<0.04). There was no significant change in osmolality over time. Mouse embryo development was significantly reduced in media with elevated osmolality (>310mOsm/kg; P<0.05). Procedures in the IVF laboratory can alter osmolality and impact embryo development.Common IVF laboratory procedures can unexpectedly alter growth conditions and impact subsequent embryo development. A media characteristic that may be altered is osmolality. The effect of different microdrop preparation conditions on media osmolality was examined. Microdrops of various volumes were prepared under different laboratory conditions. Drops were pipetted directly onto a dish and covered by oil (standard method) or placed in the dish, overlaid with oil before removing the underlying media and replaced with fresh media (wash method). Drops were made at 23°C or 37°C and were prepared in a hood with or without airflow. Media osmolality was assessed at 5min and 24h. The biological significance of the osmolality increase was demonstrated by culturing 1-cell mouse embryos in media of various osmolalities. Reduced drop volume, increased temperature and standard method were all associated with a significant increase in media osmolality at both 5min and 24h. Airflow alone had no significant impact. Additionally, 10 and 20μl volume, 37°C and standard method were associated with significant increases in media osmolality, compared with controls. There was a significant interaction effect between airflow, decreased volume, increased temperature and standard method, which caused an increase in osmolality (∼40mOsm) compared with controls. There was no significant change in media osmolality over time. Mouse embryo development was significantly reduced in media with osmolality 310 and 330mOsm. Environmental conditions in which microdrops are prepared can result in harmful media osmolality shifts. Therefore, laboratory protocols should be adjusted.

Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles

2011-10-31

Abstract: In conventional IVF cycles with total fertilization failure, rescue intracytoplasmic sperm injection (ICSI) performed 24h after insemination has yielded poor results. However, when ICSI is used, total fertilization failure is a rare event. The aim of the present study is to investigate the degree of sperm contribution to fertilization failures using the egg-sharing model in oocyte donor cycles. The study included only the oocyte donor cycles of sibling oocytes with total fertilization failure in at least one of the matched recipients. Oocytes from 49 oocyte donor cycles were equally shared among 98 recipients undergoing conventional IVF. Due to total fertilization failure in half of the recipients, rescue ICSI was carried out. Compared with the conventional IVF only group, the rescue ICSI group had a lower pregnancy rate (30.61% versus 71.43%), clinical pregnancy rate (28.57% versus 67.35%) and ongoing pregnancy rate (28.57% versus 63.27%) (all P<0.01). Cryptic sperm defects in apparently normal spermatozoa may be the cause of total fertilization failure, indicating the need for simple routine tests to detect them.Conventional IVF is an insemination method for couples with a normal semen analysis and no history of low fertilization, antisperm antibodies or poor sperm survival. This technique involves the addition of processed spermatozoa to the eggs in a culture dish, allowing the natural process of sperm selection to take place. Failures of fertilization in IVF and the attempts to rescue by intracytoplasmic sperm injection (ICSI) the oocytes that remained unfertilized 24h after conventional IVF have, in general, poor results. However, when the ICSI technique is used, total fertilization failure is a rare event. The aim of the present study was to investigate the degree of sperm contribution to fertilization failures using the egg-sharing model in oocyte donor cycles, in which the oocytes from one woman are shared between two recipients. Oocytes from 49 oocyte donor cycles were equally shared among 98 recipients undergoing conventional IVF. Due to total fertilization failure in half of the recipients, rescue ICSI was carried out. Compared with the conventional IVF only group, the rescue ICSI group had lower pregnancy rate (30.6% versus 71.4%), clinical pregnancy rate (28.6% versus 67.3%) and ongoing pregnancy rate (28.6% versus 63.3%) (all P<0.01). Cryptic sperm defects in apparently normal spermatozoa may be the cause of total fertilization failure, indicating the need for simple routine tests to detect them.

Follicular and endocrine profiles associated with different GnRH-antagonist regimens: a randomized controlled trial

2011-11-07

Abstract: This trial assessed the impact of early initiation of gonadotrophin-releasing hormone (GnRH) antagonist on follicular and endocrine profiles compared with the fixed GnRH-antagonist protocol. Eighty-five oocyte donors were randomized to GnRH antagonist starting in the mid-luteal phase of the prestimulation cycle (degarelix-ML group), on stimulation day 1 (early follicular phase, degarelix-EF group) or day 6 (fixed protocol) (mid-follicular phase, ganirelix-MF group). Subjects in the degarelix-EF and ganirelix-MF groups received placebo in the prestimulation cycle. At start of stimulation, serum concentrations of FSH (4.6±2.3 versus 6.0±1.8IU/l), LH (2.7±1.4 versus 4.7±1.9IU/l) and oestradiol (87±35 versus 129±50pmol/l) were markedly lower (P<0.001) in the degarelix-ML group than in the placebo group. The coefficients of variation of follicle size (36.7±5.5% versus 39.2±9.4%) were not significantly different. No differences in endometrial histology, embryo quality and pregnancy rates in recipient cycles were observed between the regimens. In conclusion, early administration of GnRH antagonist altered the endocrine profile without modifying the follicular synchrony for the majority of subjects. Whether patients with a more heterogeneous follicle size at start of stimulation may benefit from an earlier intervention remains to be proven.The use of gonadotrophin-releasing hormone (GnRH) antagonist protocols in patients undergoing IVF, in which the antagonist is administered from stimulation day 5 or 6 (fixed protocol), has been associated with more heterogeneous follicle growth. Also, the hormone environment at start of stimulation in the GnRH-antagonist cycle may influence clinical response and outcome. The present trial assessed the impact of earlier start of GnRH-antagonist treatment on follicular and hormone profiles compared with the fixed protocol. Eighty-five oocyte donors were randomized to GnRH antagonist starting in the mid-luteal phase of the menstrual cycle prior to stimulation, on stimulation day 1 or on stimulation day 6 (fixed protocol). Subjects starting GnRH antagonist treatment on stimulation days 1 or 6 received placebo in the prestimulation cycle. At start of stimulation, serum concentrations of the hormones FSH, LH and oestradiol were markedly lower in the GnRH-antagonist group than in the placebo group. The follicle size heterogeneity was not different between the two groups. No differences in uterine histology, embryo quality and pregnancy rates in recipient cycles were observed between the regimens. The results indicate that the general IVF population will not benefit from an early start of GnRH-antagonist treatment. Whether patients with a poor response in a previous cycle, possibly due to a more heterogeneous follicle size at the start of stimulation, may benefit from an earlier start remains to be proven.

Standardization of catheter load speed during embryo transfer: comparison of manual and pump-regulated embryo transfer

2011-11-11

Abstract: The position of transfer air bubbles after embryo transfer is related to the pregnancy rate. With the conventional manual embryo-transfer technique it is not possible to predict the final position of the air bubbles. This position mainly depends on the catheter load speed at transfer (injection speed), a parameter that remains uncontrollable with the conventional technique even after standardization of the protocol. Therefore, the development of an automated device that generates a standardized injection speed is desirable. This study aimed to examine the variation in injection speeds in manual embryo transfer and pump-regulated embryo transfer (PRET). Seven laboratory technicians were asked to perform simulated transfers using the conventional embryo-transfer technique. Their injection speeds were compared with that of a PRET device. The results indicate that in manually performed transfers, even after standardization of the protocol, there is still a large variation in injection speed, while a PRET device generates a reliable and reproducible injection speed and therefore brings new possibilities for further standardization of the embryo-transfer procedure. Future research should reveal whether these experiments mimic real clinical circumstances and if a standardized injection speed results in more exact positioning of the transferred embryos and therefore higher pregnancy rates.The position of transfer air bubbles after embryo transfer is related to the pregnancy rate. With the currently used embryo-transfer technique, in which embryos are transferred manually with a syringe, we are not able to predict the final position of the air bubbles. This position depends on the injection speed of the syringe, which remains uncontrollable in embryo transfers that are performed manually even after standardization of the protocol. Therefore we developed an automated device that generates a reliable and reproducible injection speed, a pump-regulated embryo transfer (PRET) device. This study aimed to examine the variation in injection speeds in transfers performed manually and with the PRET device. Our results indicate that in manually performed transfers, even after standardization by protocol, there is still a large variation in injection speed and that the PRET device generates a reliable and reproducible injection speed and therefore brings new possibilities for further standardization of the embryo-transfer procedure. Future research should reveal whether our experiments mimic real clinical circumstances and if a standardized injection speed results in more exact positioning of the transferred embryos and therefore higher pregnancy rates.

Ovarian stimulation and intrauterine insemination in women aged 40years or more

2011-11-21

Abstract: Fertility decreases with advancing age. This study retrospectively reviewed the results of ovarian stimulation and intrauterine insemination (IUI) in women ⩾40years old with diminished ovarian reserve or unexplained infertility who underwent treatment with ovarian stimulation/IUI with clomiphene citrate or gonadotrophin and compared them with the results of IVF and in-vitro maturation (IVM) treatments. The main outcome measures were pregnancy and live-birth rates. The profiles of the patients in ovarian stimulation, IVM and IVF groups were comparable. There were no clinical pregnancies in the clomiphene citrate and IVM groups. The clinical-pregnancy rates in the gonadotrophin and IVF groups were 2.6% and 16.9% and the live-birth rates were 2.6% and 13.7%, respectively. Compared with ovarian stimulation, IVF is most effective for women aged 40years or more. Attempting success with ovarian stimulation or IVM will delay conception unnecessarily.Fertility decreases with advancing age. The purpose of our study was to retrospectively review the results of ovarian stimulation and intrauterine insemination (IUI) in women aged ⩾40years with diminished ovarian reserve or unexplained infertility who underwent treatment with ovarian stimulation/IUI with clomiphene citrate or gonadotrophin and to compare them with those of IVF and in-vitro maturation (IVM) treatments. The main outcome measures were pregnancy and live-birth rates. The profiles of the patients in the ovarian stimulation, IVM and IVF were comparable. There were no clinical pregnancies in the clomiphene citrate and IVM groups. The clinical pregnancy in the gonadotrophin and IVF groups were 2.6% and 16.9% and the live-birth rates were 2.6% and 13.7%, respectively. Compared with ovarian stimulation, IVF is the most effective treatment for women aged ⩾40years. Attempting success with ovarian stimulation or IVM will delay conception unnecessarily.

Neurotrophins (BDNF and NGF) in follicular fluid of women with different infertility diagnoses

J.C. Sadeu, A.M.C.M. Doedée, M.S. Neal, E.G. Hughes, W.G. Foster — 2011-11-28

Abstract: Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are intra-ovarian signalling peptides that are important in follicle development and oocyte maturation. In the ovary, neurotrophin expression is regulated by gonadotrophins. Therefore, this study postulates that aetiology of infertility will affect follicular-fluid BDNF and NGF concentrations. Follicular fluid from the first follicle aspirated from 190 infertile women attending a university-affiliated fertility programme (McMaster University and ONE Fertility, Burlington, Ontario) was collected between February 2004 and November 2010. The relationship between follicular-fluid BDNF and NGF concentration and age, day-3 FSH and peak serum oestradiol concentrations and antral follicle count was determined. Participants were aged between 24 and 44years (mean±SEM, 35.2±0.3years) of age. The median concentrations of BDNF and NGF in the follicular fluid was 19.4pg/ml and 344.6ng/ml, respectively. The concentrations of BDNF and NGF were significantly related (P=0.028) but only the BDNF concentration was significantly higher (P<0.05) in women with unexplained infertility compared with other causes of infertility. It is concluded that, apart from unexplained infertility, the underlying cause of infertility did not affect ovarian output of BDNF and NGF in response to ovulation induction.Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are proteins that are produced in the brain, where they play a role in the development and survival of brain cells. Emerging evidence has shown that these proteins are also made in the ovary, where they are thought to be important in the recruitment and development of a woman’s oocytes (eggs). In the ovary, the hormones responsible for controlling the production of female hormones, such as oestradiol, are believed to also regulate the production of BDNF and NGF. Therefore, we proposed that the concentrations of these proteins in the fluid surrounding the developing egg (follicular fluid) could be affected by the underlying causes of infertility. Therefore we measured the concentration of these proteins in the follicular fluid collected from 190 infertile women attending a university-affiliated fertility programme (McMaster University and ONE Fertility, Burlington, Ontario). In this study, the concentrations of BDNF and NGF were significantly related but only the BDNF concentration was significantly higher in women with unexplained infertility compared with other causes of infertility. We conclude that, apart from unexplained infertility, the underlying cause of infertility did not affect ovarian output of these proteins.

Clinical validation of a closed vitrification system in an oocyte-donation programme

2011-11-04

Abstract: Controversy exists about the risk of microbiological contamination from direct contact with unsterile liquid nitrogen during oocyte vitrification. The aim of this observational study was to evaluate the effectiveness of oocyte vitrification using a high-security closed vitrification system in a donation programme. Oocyte vitrification was performed using CBS High Security closed straws (Cryo Bio System) with DMSO/ethylene glycol/sucrose as the cryoprotectant (Irvine Scientific freeze kit). A total of 123 vitrified metaphase-II oocytes were warmed in 20 recipient cycles (6.2 warmed oocytes per recipient); of these, 111 oocytes (90.2%) survived vitrification and warming. All surviving oocytes were microinjected and 86 (77.5%) were normally fertilized, of which 53 (61.6%) developed up to good-quality day 3. Ten embryo transfers resulted in a clinical pregnancy (50.0%) and an ongoing clinical pregnancy rate of 45%. Five revitrified embryos were warmed in three warming cycles (survival rate 100%). These transfers resulted in an additional ongoing twin pregnancy, leading to a cumulative ongoing pregnancy rate per patient of 50% (10/20). The ongoing implantation rate per warmed oocyte and per injected oocyte was 10.6% (13/123) and 11.7% (13/111). The present data demonstrate that oocyte vitrification using a closed vitrification device yields excellent oocyte survival, fertilization and embryo development.Progress in oocyte vitrification has led to successful implementation of the technique for different clinical applications. These excellent results have been achieved with open carriers, allowing direct contact of the oocyte with the liquid nitrogen. However, controversy exists about the risks of microbiological contamination in case of direct contact with the unsterile liquid nitrogen. This validation trial aimed to evaluate oocyte vitrification using a high-security closed vitrification system in a donation programme. In this study, a total of 123 vitrified metaphase-II oocytes were warmed in 20 recipient cycles resulting in 111 surviving oocytes (90.2%). Intracytoplasmic sperm injection resulted in 86 (77.5%) 2-pronuclear embryos, of which 53 (61.6%) developed to good quality by day 3. Ten embryo transfers resulted in a clinical pregnancy (50.0%) and an ongoing clinical pregnancy rate of 45%. The ongoing implantation rate including the outcome of revitrified embryos per warmed oocyte, and per injected oocyte is 10.6% (13/123) and 11.7% (13/111). The present data demonstrate that oocyte vitrification using a closed vitrification device yields excellent oocyte survival, fertilization and embryo development comparable to open vitrification. The evaluated closed vitrification device guarantees aseptic vitrification without requiring sterilized liquid nitrogen or the addition of a sealed container for storage.

Oocyte vitrification technology has made egg-sharing donation easier in China

2011-11-14

Abstract: When infertile women undergoing IVF or intracytoplasmic sperm injection (ICSI) have more than 20 mature oocytes retrieved, at least 15 oocytes are inseminated by their husband’s spermatozoa. The extra oocytes are cryopreserved by vitrification. If the patients became pregnant and have healthy live births, the patients are encouraged to donate their remaining cryopreserved oocytes. Forty-seven egg-sharing donors were recruited after having normal deliveries and they donated their remaining oocytes, totalling 395 cryopreserved oocytes, to 75 recipients. The survival rate of vitrified–warmed oocytes was 83.0%. Following insemination by ICSI, the fertilization and cleavage rates were 83.8% and 89.8%, respectively. Out of 75 recipients, 71 recipients completed the treatment cycles and 30 of them became pregnant with clinical pregnancy and implantation rates of 42.3% and 25.5%, respectively. The birthweight of the new-born infants (22 from singleton and two from one set of twins) were 3344.5±669.1g and 2425.0±742.5g, respectively. No birth defects were observed for the live births. These results indicate that oocyte vitrification is an effective methodology for an egg-sharing donation programme, with acceptable pregnancy and implantation rates.This study demonstrated that the strategy of egg-sharing donation with oocyte vitrification is a practical and efficient programme in China, especially with the strict regulations controlling egg-sharing donation. When infertile women undergoing IVF or intracytoplasmic sperm injection (ICSI) have more than 20 mature oocytes retrieved, at least 15 oocytes are inseminated by their husband’s spermatozoa. The extra oocytes are cryopreserved by vitrification. If the patients become pregnant and have healthy live births, the patients are encouraged to donate their remaining cryopreserved oocytes. Forty-seven egg-sharing donors were recruited after having healthy deliveries and they donated their remaining oocytes, in total 395 cryopreserved oocytes to 75 recipients. The survival rate of vitrified–warmed oocytes was 83.0%. Following insemination by ICSI, the fertilization and cleavage rates were 83.8% and 89.8% respectively. Out of 75 recipients, 71 recipients completed their treatment cycle and 30 of them became pregnant (42.3% clinical pregnancy rate and 25.5% implantation rate). The birthweight of new-born infants for 22 singletons and two babies from one set of twins were in the normal range. No birth defects were observed for those live births. These results indicate that oocyte vitrification is an effective methodology for an egg-sharing donation programme, and gives acceptable pregnancy and implantation rates.

Spontaneous LH surges prior to HCG administration in unstimulated-cycle frozen–thawed embryo transfer do not influence pregnancy rates

E.R. Groenewoud, B.J. Kollen, N.S. Macklon, B.J. Cohlen — 2011-11-11

Abstract: LH surges are the start of a period of optimal endometrial receptivity. Missing these surges in an unstimulated-cycle frozen–thawed embryo transfer (FET) based on ultrasound alone might lead to incorrect timing of embryo transfer. This prospective, non-randomized trial established the incidence and effect of spontaneous LH surges on ongoing pregnancy rates and assessed the use of ultrasound without LH monitoring in planning FET. All patients undergoing unstimulated-cycle FET in the study centre over a 2-year period were included in this analysis (n=233). All patients had regular menstrual cycles. Serum LH analysis took place before human chorionic gonadotrophin administration. The main outcome measure was ongoing pregnancy. LH surges occurred in over half of patients. Overall pregnancy rate was 34.3%. Relative risks for ongoing pregnancy for cycles with or without a spontaneous LH surge were not significantly different (ongoing pregnancy rate 33.4% versus 34.8%; RR 1.02, 95% CI 0.7–1.5). Based on these results, it was concluded that LH surges ⩾10 IU/l occurred in over 50% of patients, but LH surges demonstrated no significant effect on pregnancy rates. Single LH determination prior to ovulation induction in unstimulated-cycle FET does not seem to have added clinical value.LH surges are the start of a period of fertility. Missing these surges in natural-cycle frozen–thawed embryo transfer (FET) based on ultrasound alone might lead to incorrect timing of thawing and transferring. This can subsequently lead to diminished pregnancy rates. In this trial we established both the incidence and effect of these LH surges on ongoing pregnancy rates and assessed the use of ultrasound without LH monitoring in planning natural-cycle FET. Over a 2-year period, all patients undergoing natural-cycle FET in our centre were included in this analysis (n=233). All patients had a regular menstrual cycle. Analyses of the LH concentration took place before ovulation induction with human chorionic gonadotrophin in natural-cycle FET. The main outcome measure was ongoing pregnancy. LH surges occurred in over half of all patients. The overall pregnancy rate was 34.3%. No difference was found in pregnancy rates between patients with and without an LH surge. Based on these results, we concluded that LH surges occurred in over 50% of all patients, but these surges demonstrated no significant effect on pregnancy rates. Regular ultrasound evaluation of the dominant follicle alone seems to be an accurate method to plan natural-cycle FET.

Apolipoprotein E-containing HDL-associated platelet-activating factor acetylhydrolase activities and malondialdehyde concentrations in patients with PCOS

Ping Fan, Hongwei Liu, Ying Wang, Feng Zhang, Huai Bai — 2011-10-31

Abstract: PAF and PAF-like oxidized phospholipids hydrolysed by platelet-activating factor (PAF) acetylhydrolase (AH) are potent lipid mediators involved in inflammation and atherosclerosis. Apolipoprotein (apo) E-containing high-density lipoprotein (HDL) has antioxidant, anti-inflammatory and anti-atherogenic properties. The study investigated apoE-containing HDL-associated PAF-AH (HDL-PAF-AH) and total (apoE-containing+apoE-poor) HDL-PAF-AH activities as well as malondialdehyde (MDA) concentration in 291 patients with polycystic ovary syndrome (PCOS) using the Rotterdam consensus criteria and 281 control women. Compared with the control women, patients with hyperandrogenism+oligo/anovulation+polycystic ovaries (PCO) or hyperandrogenism+PCO had lower total, apoE-containing and apoE-poor HDL-PAF-AH activities, while those with oligo/anovulation+PCO showed decreased total and apoE-poor HDL-PAF-AH activities. Other factors including insulin resistance and obesity in PCOS had the adverse effects associated with the HDL-PAF-AH activities. Serum MDA concentration was associated with PCOS, hyperandrogenism, insulin resistance and hypertriglyceridaemia in patients with PCOS. Decreased total and apoE-containing HDL-PAF-AH activities and increased serum MDA concentration may contribute to the pathogenesis of PCOS and potentially link to related complications responsible for oxidative stress and inflammation such as an increased risk for type 2 diabetes mellitus and/or future cardiovascular diseases in PCOS patients.Platelet-activating factor (PAF) acetylhydrolase (PAF-AH) hydrolyses and inactivates PAF and PAF-like oxidized phospholipids that are potent lipid mediators involved in inflammation and atherosclerosis. Apolipoprotein (apo) E-containing high-density lipoprotein (HDL) has antioxidant, anti-inflammatory and anti-atherogenic properties. The study investigated apoE-containing HDL-associated PAF-AH (HDL-PAF-AH) activities, total (apoE-containing+apoE-poor) HDL-PAF-AH activities as well as malondialdehyde (MDA) concentration in 291 patients with polycystic ovary syndrome (PCOS) using the Rotterdam consensus criteria and 281 control women. Our results reported for the first time, as far as is known, that the apoE-containing HDL has a relatively high PAF-AH activity and accounts for about 50% of total HDL-PAF-AH activity in both the patient and control groups. Compared with the control women, the patients with hyperandrogenism+oligo/anovulation+polycystic ovaries (PCO) or hyperandrogenism+PCO had lower total, apoE-containing and apoE-poor HDL-PAF-AH activities, while those with oligo/anovulation+PCO showed decreased total and apoE-poor HDL-PAF-AH activities. Other factors, including insulin resistance and obesity in PCOS, had adverse effects associated with the HDL-PAF-AH activities. Serum MDA concentration was associated with PCOS, hyperandrogenism, IR and hypertriglyceridaemia in patients with PCOS. Since HDL-PAF-AH activity has been consistently shown to play an anti-inflammatory and anti-atherogenic role, the decreased total and apoE-containing HDL-PAF-AH activities and increased serum MDA concentration may contribute to the pathogenesis of PCOS and potentially link to related complications responsible for oxidative stress and inflammation, such as an increased risk for type 2 diabetes mellitus and/or future cardiovascular diseases in PCOS patients.

Sperm parameters and male fertility after bariatric surgery: three case series

2011-11-04

Abstract: Recent studies have underlined the impact of obesity on sperm parameters, but very few data are available on the effect of weight loss on male fertility. This article reports the case series of three male patients who underwent rapid and major weight loss following bariatric surgery and the consequences of this surgery on semen parameters and fertility. A severe worsening of semen parameters was observed during the months after bariatric surgery, including extreme oligoasthenoteratozoospermia, but azoospermia was not observed. This effect may hypothetically be the result of two opposite mechanisms: (i) the suppression of the deleterious effects of obesity; and (ii) the negative impact of both nutritional deficiencies and the release of toxic substances. Information about potential reproductive consequences of bariatric surgery should be given to patients and sperm cryopreservation before surgery proposed. However, for one case, the alterations of spermatogenesis were reversible 2years after the surgical procedure. Finally, intracytoplasmic sperm injection with fresh spermatozoa after male bariatric surgery can be successful, as demonstrated here, where clinical pregnancies were obtained for two out of the three couples.Recent studies have underlined the impact of obesity on sperm parameters, but very few data are available on the effect of weight loss on male fertility. We here report the case series of three male patients who underwent rapid and major weight loss following obesity surgery and the consequences of this on semen parameters and fertility. A severe worsening of semen parameters was observed during the months after surgery, but an absence of spermatozoa was not observed. This effect may hypothetically be the result of two opposite mechanisms, firstly the suppression of the deleterious effects of obesity and secondly the negative impact of both nutritional deficiencies and the release of toxic substances. Information about the potential reproductive consequences of obesity surgery should be given to patients and sperm cryopreservation before surgery proposed. However, for one case, the alterations of spermatogenesis were reversible 2years after the surgical procedure. Finally, IVF with fresh spermatozoa after male bariatric surgery can be successful, as demonstrated in our case series, where clinical pregnancies were obtained for two of the three couples.

Correlation between DNA defect and sperm-head morphology

2011-10-24

Abstract: The utility of sperm DNA testing remains controversial. However, it may be helpful in couples with unexplained failures of multiple assisted reproductive techniques and/or recurrent abortions. This study analysed 10,400 spermatozoa of 26 patients for sperm-head morphology with high-magnification microscopy, DNA fragmentation and sperm chromatin decondensation. A significant negative correlation was demonstrated between sperm-parameters and abnormal sperm-head morphology as assessed by high magnification (score 0 according to this study’s classification): concentration (r=−0.41; P=0.03), motility (r=−0.42; P=0.03), morphology (r=−0.63; P=0.0008). No correlation was found with DNA fragmentation. However, the sperm chromatin-decondensation rate of score-0 spermatozoa was twice as high as the controls (19.5% versus 10.1%; P<0.0001). This observation suggests that score-0 spermatozoa should not be selected for intracytoplasmic sperm injection.We analysed 10,400 spermatozoa of 26 patients for sperm head morphology at high magnification, DNA fragmentation and sperm chromatin decondensation. We demonstrated a significant negative correlation between sperm parameters: concentration (r=–0.41; P=0.03), motility (r=−0.42; P=0.03), morphology (r=–0.63; P=0.0008) and abnormal sperm head morphology as assessed by high magnification (score 0 according our classification). No correlation was found with DNA fragmentation. The sperm chromatin decondensation rate of score-0 spermatozoa was twice as high as than in controls (19.5% versus 10.1%; P<0.0001). No significant relationship with sperm DNA fragmentation was observed. We suggest that high-magnification sperm selection could be an important step and a new tool for the clinician, who could decide to discard score-0 spermatozoa with a high risk of abnormal chromatin and select the best sperm cells for intracytoplasmic sperm injection.

Sperm FISH analysis of a 46,XY,t(3;6)(p24;p21.2),inv (8)(p11;2q21.2) double chromosomal rearrangement

2011-10-28

Abstract: A complex chromosome rearrangement (CCR) can be defined as a structural chromosomal aberration that involves at least three breakpoints located on two or more chromosomes. Highly unbalanced gametes may lead to infertility or congenital malformations. Here is reported a double rearrangement considered as the simplest possible CCR and, in a sense, not a true CCR, meiotic segregation for a 46,XY,t(3;6)(p24;p21.2),inv(8)(p11;2q21.2) male patient referred after his partner had undergone three early miscarriages. Sperm fluorescence in-situ hybridization was used to screen for translocation and inversion segregation and an interchromosomal effect (ICE) for 13 chromosomes not involved in CCR. The malsegregation rates for the reciprocal translocation and pericentric inversion were 61.2% and 1.7%, respectively. ICE analysis revealed that the observed chromosome aneuploidy rates of between 0.1% and 0.8% did not differ significantly from control values. A slight increase in cumulative ICE (P=0.049) was observed in the patient, relative to control spermatozoa (with rates of 4.6% and 3.1%). The sperm DNA fragmentation rate differed significantly from control values (5.0%; P=0.001). Reciprocal translocation had no impact on meiotic segregation of the pericentric inversion in this double rearrangement. No conclusion could be drawn regarding the impact of pericentric inversion on translocation.A complex chromosome rearrangement (CCR) can be defined as a structural chromosomal aberration that involves at least three breakpoints located on two or more chromosomes, with the exchange of genetic material. Highly unbalanced gametes may lead to infertility or congenital malformations. We report here the first known double rearrangement CCR meiotic segregation for a 46,XY,t(3;6)(p24;p21.2),inv(8)(p11;2q21.2) male patient referred to our hospital after his partner had undergone three early miscarriages. Sperm fluorescence in-situ hybridization was used to screen for translocation and inversion segregation and an interchromosomal effect (ICE) for 13 chromosomes not involved in CCR (chromosomes 7, 9, 11, 12, 13, 15, 16, 17, 18, 20, 21, X and Y). The malsegregation rates for the reciprocal translocation and pericentric inversion were 61.2% and 1.7%, respectively. An ICE analysis revealed that the observed chromosome aneuploidy rates between 0.1% and 0.8% did not differ significantly from control values (between 0.1% and 0.5%). A slight increase in cumulative ICE for the 13 chromosomes (P=0.049) was observed in the CCR patient, relative to control spermatozoa (with rates of 4.6% and 3.1%). The sperm DNA fragmentation rate was slightly higher than our laboratory’s normal cut-off value (13%) and differed significantly from controls values (5.0%; P=0.001). In agreement with previous results for isolated inversions, we conclude that reciprocal translocation had no impact on meiotic segregation of the pericentric inversion in this double rearrangement CCR. No conclusion could be drawn regarding the impact of pericentric inversion on translocation.

Sphingosine–sphingosine-1-phosphate pathway regulates trophoblast differentiation and syncytialization

Ambika T. Singh, Arunasalam Dharmarajan, Irving L.M.H. Aye, Jeffrey A. Keelan — 2011-11-04

Abstract: Sphingosine and sphingosine-1-phosphate (S1P) are involved in regulating cell differentiation. This study postulated that changes in sphingolipid biosynthesis and metabolism are important in trophoblast syncytialization and therefore examined the production, metabolism and actions of sphingosine and S1P during spontaneous trophoblast differentiation and fusion in vitro. Significant declines in intracellular sphingosine concentration (P⩽0.05) and sphingosine kinase 1 (SPHK1) expression (P⩽0.01) were observed during trophoblast syncytialization. Secreted S1P concentrations dropped steeply after 72h, before rising to basal concentrations with syncytialization. Intracellular S1P concentrations were undetectable throughout. Treating cells with exogenous sphingosine (P⩽0.01), S1P (P⩽0.001) or a specific SPHK1 inhibitor (P⩽0.05) for up to 72h in culture significantly inhibited trophoblast differentiation (measured as reduced human chorionic gonadotrophin production); effects on other biochemical and morphological markers of differentiation were absent or inconsistent. Phosphorylation of Akt, an established down-stream target of S1P that spontaneously declines with trophoblast differentiation, was markedly reduced by S1P (P⩽0.05). In conclusion, changes in the sphingosine–S1P pathway are involved in the regulation of trophoblast differentiation in term human placenta. Dysregulation of sphingolipid homeostasis could, therefore, disrupt placental formation and function with deleterious consequences for pregnancy outcome.Sphingolipids are known to be key lipid mediators, signalling molecules and regulators of cellular differentiation. We postulated that changes in sphingolipid biosynthesis and metabolism might play a role in the fusion and differentiation of placental trophoblast cells, and therefore examined the production, metabolism and actions of sphingosine and sphingosine-1-phosphate (S1P) and during spontaneous trophoblast differentiation in vitro. Significant declines in intracellular sphingosine concentrations and sphingosine kinase 1 (SPHK1) expression were observed during trophoblast differentiation and fusion. Secreted S1P concentrations dropped steeply at the peak of differentiation, before rising back to basal concentrations with syncytialization. Intracellular S1P concentrations were undetectable. Treating cells with sphingosine, S1P or a specific SPHK1 inhibitor for up to 72h in culture significantly inhibited trophoblast differentiation as assessed by some markers, but not others. Phosphorylation of Akt, a common downstream target of S1P, was markedly reduced by S1P treatment. We conclude that changes in cellular sphingosine concentration and phosphorylation are involved in the regulation of trophoblast differentiation in term human placenta. Disturbances in sphingolipid homeostasis could, therefore, disrupt placental formation and function and compromise pregnancy outcome and fetal wellbeing.

Extra-embryonic human Wharton’s jelly stem cells do not induce tumorigenesis, unlike human embryonic stem cells

2011-10-24

Abstract: Tumorigenesis is the major obstacle of tissues derived from human embryonic stem cells (ESC) and human induced pluripotent stem cell (IPSC) for transplantation therapy. This prompted a search for other sources of ESC. This study isolated and characterized stem cells from the extra-embryonic human umbilical cord Wharton’s jelly (WJSC). These cells are non-controversial, available in abundance, proliferative, multipotent and hypoimmunogenic. However, their tumorigenic potential has not been properly addressed. Their tumour-producing capabilities were compared with human ESC using the immunodeficient mouse model. Unlabelled human ESC+matrigel (2×106cells/site), labelled human WJSC (red fluorescent protein; 5×106cells/site) and unlabelled human WJSC+matrigel (5×106cells/site) were injected via three routes (s.c., i.m. and i.p.). Animals that received human ESC+matrigel developed teratomas in 6weeks (s.c. 85%; i.m. 75%; i.p. 100%) that contained tissues of ectoderm, mesoderm and endoderm. No animal that received human WJSC developed tumours or inflammatory reactions at the injection sites when maintained for a prolonged period (20weeks). Human WJSC produced increases in anti-inflammatory cytokines in contrast to human ESC, which increased pro-inflammatory cytokines. Human WJSC, being hypoimmunogenic and non-tumorigenic, have the potential for safe cell-based therapies.Human embryonic stem cells (human ESC) and human induced pluripotent stem cells (IPSC) are versatile as they can be converted to all the tissue types of the human body (pluripotency). However, a major clinical hurdle in using hESC- and IPSC-derived tissues for treating disease is their ability to produce tumours. This disadvantage prompted our search for a novel stem cell in the human Wharton’s jelly, a gelatinous substance in the umbilical cord. Human Wharton’s jelly stem cells (WJSC) are available in large numbers, can be grown easily in culture, have prolonged stem cell-like properties, can be converted into many desirable tissues and, when transplanted into diseased animal models, correct disease. However, since their ability to produce tumours has not been properly addressed, we compared their tumour-forming capabilities with human ESC in immunodeficient mice. Human ESC (2×106cells/site) and human WJSC (5×106cells/site) were injected via three routes: under the skin (subcutaneous), into muscle (intramuscular) and into the abdominal cavity (intraperitoneal). All animals that received human ESC developed tumours in 6weeks and there were no tumours in the animals receiving human WJSC, even when monitored for 20weeks. Also, the animals receiving human ESC showed immunological responses while those receiving human WJSC did not. We conclude that human WJSC have enormous potential for cell-based therapies as they do not induce tumours and are not rejected when transplanted.

Triggering with GnRH agonist in oocyte-donation cycles: oestradiol monitoring is not necessary during ovarian stimulation

2011-11-21

Abstract: This prospective observational study evaluated the efficacy and safety of oocyte-donation cycles triggered with a gonadotrophin-releasing hormone (GnRH) agonist without monitoring oestradiol concentrations during ovarian stimulation. A total of 97 oocyte donors received recombinant FSH (150–225/day) and GnRH antagonists (0.25mg/day). Oocyte maturation was triggered with 0.2mg triptorelin s.c. Donors aged 25.4±4.1years were stimulated for 8.8±0.9days and underwent 2.9±0.5 (2–4) ultrasound assessments. Total FSH dose was 1703.4±304.7IU, antagonists were administered for 4.3±1.0days, 14.7±8.8 oocytes were retrieved and there were no cases of ovarian hyperstimulation syndrome. Recipients (n=123) aged 40.3±3.4years received 10.9±4.3 oocytes, 88.7% of which were metaphase II. Intracytoplasmic sperm injection fertilization rate was 79% and 2.18±0.6 (1–3) embryos were transferred. The pregnancy, clinical pregnancy and twin pregnancy rates were 64.2%, 57.7% and 19.7%, respectively. In conclusion, given the high efficacy and safety of the GnRH-antagonist protocol triggered with a GnRH agonist, the monitoring of oestradiol concentrations is not necessary. Ultrasound monitoring is enough for an adequate follow up of the stimulation cycle in oocyte donors.

Absence of SYCP3 mutations in women with recurrent miscarriage with at least one trisomic miscarriage

Courtney W. Hanna, John D. Blair, Mary D. Stephenson, Wendy P. Robinson — 2011-11-04

Abstract: Mutations within the coding regions of the synaptonemal complex gene SYCP3 have previously been reported in women with recurrent miscarriage. The present study found no mutations in any of the coding exons or the intron/exon boundaries among 50 recurrent miscarriage patients with at least one documented trisomic miscarriage, suggesting that mutations in SYCP3 do not contribute significantly to risk for recurrent miscarriage through maternal meiotic nondisjunction.

Response: Low-intensity IVF: real progress?

John Zhang, Sherman Silber — 2011-11-28

We are pleased to have been invited to respond to the Commentary ‘Low-intensity IVF: real progress?’ by so as to highlight our differing positions. The results of most IVF series, including yearly Society for Assisted Reproductive Technologies reports and European Society of Human Reproduction and Embryology reports, are not randomized control trials. We attempted to make it clear in our report that our study was not a randomized controlled trial. However, it is totally true that results with mini-IVF, if performed to high standards as described in our report, are indeed comparable to those achieved with conventional ovarian stimulation. To us, it also seems obvious that it is safer, cheaper and easier on patients. What also seems to us obvious is that the authors of the Commentary feel uncomfortable about the inevitability of greater adoption of the practice of elective single embryo transfer, even though we all know that this is the future. Our separate anecdotal experience in St Louis also shows results with mini-IVF that are comparable to conventional IVF, and even superior in some types of patients, such as older women or women with low ovarian reserve. Finally, we are indeed conducting a randomized control trial, with outside monitoring, comparing ‘mild’ stimulation to ‘conventional’ stimulation, as we promised in our article.

Reproductive BioMedicine Online

Contents

The Edwards and Steptoe Research Trust: commemorating the two great pioneers of our field and supporting their aspirations

Useful lessons from China?

Professor Edwin Carlyle (Carl) Wood AC, CBE, FRCS, FRCOG, FANZCOG

The luteal phase after GnRH-agonist triggering of ovulation: present and future perspectives

Microdrop preparation factors influence culture-media osmolality, which can impair mouse embryo preimplantation development

Cryptic sperm defects may be the cause for total fertilization failure in oocyte donor cycles

Follicular and endocrine profiles associated with different GnRH-antagonist regimens: a randomized controlled trial

Standardization of catheter load speed during embryo transfer: comparison of manual and pump-regulated embryo transfer

Ovarian stimulation and intrauterine insemination in women aged 40years or more

Neurotrophins (BDNF and NGF) in follicular fluid of women with different infertility diagnoses

Clinical validation of a closed vitrification system in an oocyte-donation programme

Oocyte vitrification technology has made egg-sharing donation easier in China

Spontaneous LH surges prior to HCG administration in unstimulated-cycle frozen–thawed embryo transfer do not influence pregnancy rates

Apolipoprotein E-containing HDL-associated platelet-activating factor acetylhydrolase activities and malondialdehyde concentrations in patients with PCOS

Sperm parameters and male fertility after bariatric surgery: three case series

Correlation between DNA defect and sperm-head morphology

Sperm FISH analysis of a 46,XY,t(3;6)(p24;p21.2),inv (8)(p11;2q21.2) double chromosomal rearrangement

Sphingosine–sphingosine-1-phosphate pathway regulates trophoblast differentiation and syncytialization

Extra-embryonic human Wharton’s jelly stem cells do not induce tumorigenesis, unlike human embryonic stem cells

Triggering with GnRH agonist in oocyte-donation cycles: oestradiol monitoring is not necessary during ovarian stimulation

Absence of SYCP3 mutations in women with recurrent miscarriage with at least one trisomic miscarriage

Response: Low-intensity IVF: real progress?