Reproductive BioMedicine Online

2012-05-01

The uses and abuses of bibliometrics

Martin H. Johnson, Jacques Cohen, Gedis Grudzinskas — 2012-05-01

Bibliographic databases provide us with easy access to many more journals than was possible when each of us editors started in science. We well remember the treks around libraries, the frustration of the missing volumes and the broken photocopier, the ‘reprint request’ post cards sent and received, and the bundling up of hard copy papers for posting to our colleagues. Now, at the press of a button, papers can be located, paid for (unless freely accessible), down loaded, filed – and sometimes even read! However, with this easy access to databases and papers come problems: notably the increased risk of deliberate or accidental plagiarism () and the fact of information overload. This latter problem has resulted in what can be seen as an abreaction: the entrenchment of the ‘prestige journal’ into which a young scientist must get their paper come what may – much of the data incomprehensibly compacted, and figures often too small or cropped to be of evidential value. These ‘prestige’ journals build and thrive financially through the increased significance of a development complementary to bibliographic databases: the bibliometric analysis.

Can oestradiol pretreatment be used to reliably avoid weekend oocyte retrievals?

G. Griesinger, E.M. Kolibianakis — 2012-02-06

Abstract: Scheduling the initiation of ovarian stimulation in a gonadotrophin-releasing hormone (GnRH)-antagonist protocol by sex steroid pretreatment has been suggested as a means to reduce the incidence of oocyte retrievals during weekends. The rationale is that by manipulating the initiation of gonadotrophin stimulation, Thursday or Friday will be avoided as days on which triggering of final oocyte maturation will be performed and thus weekend oocyte retrievals will not occur. Apparently, the assumption behind such an approach is that duration of stimulation is homogenous enough to serve this purpose reliably. However, existing data suggest that large inter-individual variation exists in the duration of gonadotrophin stimulation required to reach predefined criteria for triggering final oocyte maturation, regardless of whether stimulation was initiated with spontaneous menstruation or after pretreatment with sex-steroids. Therefore, it is highly unlikely that any type of pretreatment aiming to allow initiation of stimulation on a certain day will result in avoidance of weekend oocyte retrievals, when predefined criteria for triggering final oocyte maturation are used.

Lynette Scott (1954–2012): a force of nature

2012-03-01

Lynette Scott died suddenly on 2 February 2012 in Massachusetts, following pulmonary embolism. Her untimely death will ultimately leave a void with those who were fortunate to know her personally and those who worked with her professionally. Lynette is survived by her husband, Alan and sons Nicholas 22 and Joseph 18.

Ovarian biopsy has no role as a routine diagnostic test of ovarian reserve: a systematic review

Ricky Kwok, Neil P. Johnson — 2012-02-02

Abstract: Ovarian reserve describes a woman’s reproductive potential and there are a variety of tests for this. This reflects the lack of a gold standard and the lack of a single test that provides sufficient accuracy. Ovarian biopsy has been proposed as a potential tool for assessing the ovarian reserve and therefore the ability or inability for a woman to bear a child with or without treatment. The literature assessing the diagnostic accuracy of ovarian biopsy as a test of ovarian reserve for predicting fertility outcomes (live birth rate, ongoing pregnancy, clinical pregnancy, biochemical pregnancy, embryos available, oocytes retrieved or cancelled cycles) was systematically reviewed. There were no studies identified that assessed the diagnostic accuracy of ovarian biopsy for predicting fertility outcomes but a number of studies provided evidence that ovarian follicles are distributed unevenly and randomly throughout the ovarian cortex. This leads to sampling error when ovarian biopsy is used to sample the ovarian reserve. It is concluded that ovarian biopsy should not be used as a test of ovarian reserve.Every woman is born with a limited number of eggs, and as women age, their ability to have children eventually decreases to zero due to the loss of these eggs. Ovarian reserve is a term that describes the remaining pool of eggs. Testing for ovarian reserve enables one to predict the ability or inability of a woman to have a child and how long a woman may be able to defer having a child. This is of importance as social trends in developed countries are leading to increasing numbers of women who have children at later stages of their lives. One test of ovarian reserve is an ovarian biopsy, where a woman undergoes an operation to have a small piece of her ovary removed. The number of eggs in this removed piece is counted, and from this, the number of eggs in the whole ovary can be estimated and calculated. We reviewed the literature for all studies that examined the accuracy of ovarian biopsy to predict fertility outcomes but found no studies that answered this question. A number of studies have found that eggs are distributed unevenly and randomly in an ovary. As such, the number of eggs in an ovarian biopsy sample does not represent the actual number of eggs remaining in the ovary. Because of this and the risks inherent in the operation to perform this test, we believe that ovarian biopsy should not be used as a test of ovarian reserve.

The eutopic endometrium in endometriosis: are the changes of clinical significance?

Ivo Brosens, Jan J. Brosens, Giuseppe Benagiano — 2012-02-02

Abstract: The eutopic endometrium in women suffering from endometriosis is different in many ways from that of healthy controls. Both proliferative and secretory eutopic endometria exhibit changes in endometriosis with heterogeneous responses. In addition, nerve fibres appear in the endometrium and myometrium of these women. The endometrium is a rich source of pro-angiogenic factors and vascular events are often disrupted in endometriosis with an overall increase in angiogenesis. A number of investigations have shown that endometriosis is likely the most common cause of endometrial receptivity defects. Endometriosis is also associated with relative 17β-hydroxysteroid dehydrogenase type II deficiency and these molecular aberrations indicate that local oestrogen production sustains ectopic implants. Recently it has been shown that endometriosis, as a chronic inflammatory disorder, disrupts co-ordinated progesterone response throughout the reproductive tract, including the endometrium, leading to a condition of ‘progesterone resistance’. Investigators have searched for biomarkers of endometriosis, but these investigations are fraught with methodological difficulties. In conclusion, molecular phenotyping of the endometrium is changing the disease paradigm, from being foremost an oestrogen-dependent disease to a disorder characterized primarily by progesterone resistance.In recent years, research on the pathogenesis of endometriosis has been focused on alterations in the uterus and particularly the eutopic endometrium. The eutopic endometrium in women suffering from endometriosis is different in many ways from that of healthy controls. Both proliferative and secretory eutopic endometria exhibit changes in endometriosis with heterogeneous responses. The endometrium is a rich source of pro-angiogenic factors and vascular events are often disrupted in endometriosis with an overall increase in angiogenesis. A number of investigations have shown that endometriosis is likely the most common cause of endometrial receptivity defects. Recently, it has been shown that endometriosis, as a chronic inflammatory disorder, disrupts co-ordinated progesterone response throughout the reproductive tract, including the endometrium, leading to a condition of ‘progesterone resistance’. Investigators have searched for biomarkers of endometriosis, but these investigations are fraught with methodological difficulties. In conclusion, molecular phenotyping of the endometrium is changing the disease paradigm; from being foremost an oestrogen-dependent disease to a disorder characterized primarily by progesterone resistance.

Can the fall in serum FSH during coasting in IVF/ICSI predict clinical outcomes?

2012-02-16

Abstract: This retrospective cohort study determined whether the total falls in serum FSH and oestradiol concentrations from start to end of coasting in IVF/intracytoplasmic sperm injection could predict clinical outcomes. Ninety-nine cycles, with gonadotrophin-releasing hormone-agonist down-regulation where coasting with serial serum oestradiol and FSH monitoring was adopted due to risk of severe ovarian hyperstimulation syndrome, were consecutively included. The primary clinical outcome was live-birth rate (LBR); other outcomes measured were number of oocytes retrieved and fertilization, implantation and clinical pregnancy rates. LBR for FSH fall>10IU/l compared with 5–10 and<5IU/l were 45.4% versus 22.0% and 25.0%, respectively. Mean serum FSH fall was similar with and without live birth (8.4±6.2 versus 7.3±5.0IU/l) as were mean oestradiol and FSH concentrations on HCG administration, oestradiol fall, percentage fall in FSH/oestradiol and duration of coasting. None of the variables efficiently predicted live birth on regression analysis. The AUC of FSH fall was 0.53 at 11.0IU/l. Basal FSH, starting and total gonadotrophin dose and duration of coasting were positively correlated with FSH fall. A potentially clinically important association between live birth and FSH fall during coasting was apparent, which requires further evaluation.The purpose of this retrospective cohort study was to determine whether the magnitude of fall in the serum FSH and oestradiol concentrations from start to end of coasting in IVF/intracytoplasmic sperm injection cycles could predict the clinical outcomes. Gonadotrophin-releasing hormone-agonist down-regulated cycles (n=99), where coasting with serial serum oestradiol and FSH monitoring was adopted due to risk of ovarian hyperstimulation, were consecutively included. Live birth was the primary clinical outcome measured; number of oocytes retrieved and fertilization, implantation and clinical pregnancy rates were the other outcomes examined. Live-birth rate tended to be high when FSH fall was >10IU/l, compared with 5–10IU/l and <5IU/l, although not statistically significantly. Mean serum FSH fall were similar in live-birth and no-live-birth cycles (8.4±6.2 versus 7.3±5.0) as were mean oestradiol and FSH concentrations on hCG administration, oestradiol fall, percentage fall in FSH and oestradiol and duration of coasting. None of the variables efficiently predicted live birth. The area under the curve of FSH fall was 0.53. FSH fall of <11.0IU/l was found to be more likely to predict negative outcome (specificity 84.72%) than predicting positive outcome when FSH fall was >11IU/l (sensitivity 34.48%). Women’s basal FSH, starting and total gonadotrophin dose of ovarian stimulation and duration of coasting had direct positive correlation with the magnitude of FSH fall. A potentially clinically important rise in live birth in association with greater FSH fall during coasting was apparent, which requires further evaluation.

Effect of HCG-day serum progesterone and oestradiol concentrations on pregnancy outcomes in GnRH agonist cycles

2012-02-16

Abstract: This study analysed the relationship between serum progesterone/oestradiol concentrations and IVF pregnancy outcomes in gonadotrophin-releasing hormone agonist protocols. A total of 2921 infertile women undergoing IVF were assigned to four groups according to serum progesterone and oestradiol concentrations on the day of human chorionic gonadotrophin (HCG) administration: group 1 (control) progesterone<3.34nmol/l and oestradiol<19,124pmol/l; group 2 (high oestradiol); group 3 (high progesterone); group 4 (high progesterone and high oestradiol). Compared with group 1, group 4 had lower clinical pregnancy and live birth rates as well as the highest ectopic pregnancy rate (29.15% versus 45.91%; 18.67% versus 34.34%; 18.10% versus 5.82%; P<0.05). Group 3 had lower clinical pregnancy and live birth rates per embryo-transfer cycle (29.78% versus 45.91%; 20.28% versus 34.34%, respectively; P<0.05). Clinical pregnancy rates were similar in frozen–thawed embryo transfers (FET) among the four groups. In conclusion, elevated progesterone was detrimental to live birth rates. High serum oestradiol concentration on HCG day did not affect the IVF pregnancy outcome. In combination with the elevated progesterone, high oestradiol concentrations had a potential negative effect. For these patients, FET should be suggested to improve the pregnancy outcomes.The aim of this study was to analyse the relationship between serum progesterone/oestradiol concentrations and IVF pregnancy outcomes in gonadotrophin-releasing hormone agonist protocols. A total of 2921 infertile women undergoing IVF were assigned to four groups according to their serum progesterone and oestradiol concentrations on the day of human chorionic gonadotrophin (HCG) administration: group 1 (control) progesterone<3.34nmol/l and oestradiol<19,124pmol/l; group 2 (high oestradiol); group 3 (high progesterone); group 4 (high progesterone and high oestradiol). Compared with group 1, patients in group 4 had lower clinical pregnancy (29.15% versus 45.91%) and live birth rates (18.67% versus 34.34%) as well as the highest ectopic pregnancy rate (18.1% versus 5.82%) (all P<0.05). Those in group 3 had lower clinical pregnancy and live birth rates per embryo transfer cycle (29.78% versus 45.91%; 20.28% versus 34.34%, respectively, P<0.05). Embryo quality appeared to be unaffected since similar clinical pregnancy rates in frozen–thawed embryo transfer (FET) cycles among the four groups. In conclusion, elevated progesterone was detrimental to live birth rates. A high serum oestradiol concentration on the day of HCG administration did not affect the IVF pregnancy outcome. In combination with the elevated progesterone and oestradiol concentrations had a potential negative effect. For these patients, FET should be suggested to improve the pregnancy outcomes.

The benefit of artificial oocyte activation is dependent on the fertilization rate in a previous treatment cycle

Markus Montag, Maria Köster, Katrin van der Ven, Ulrike Bohlen, Hans van der Ven — 2012-02-13

Abstract: Following intracytoplasmic sperm injection (ICSI), some patients present low or zero fertilization rates. Artificial oocyte activation has been proposed as a suitable means to overcome this problem. This study applied artificial oocyte activation in patient cohorts with a history of no fertilization (0%, group 1), fertilization between 1 and 29% (group 2) or fertilization between 30 and 50% (group 3) in initial ICSI cycles. In the following treatment cycles, oocytes were activated after ICSI using calcium ionophore. Fertilization, pregnancy and take-home baby rates were compared with the previous cycle without activation. In group 1, fertilization rate was 41.6%, embryos for transfer were available in 82.1% of cycles, giving a clinical pregnancy rate of 18.8% and take-home baby rate of 12.8%. In group 2, despite a lower transfer rate (87.9% versus 100%, P<0.05), there were higher fertilization and clinical pregnancy rates (44.4% versus 19.3% and 31.4% versus 12.8%, respectively, P<0.05) and take-home baby rate was 24.1% versus 12.8%. In group 3, fertilization rates differed (56.1% versus 36.8%; P<0.001) but all other parameters were similar. Artificial oocyte activation has great potential especially in patients showing compromised fertilization rates below 30% after standard ICSI.Following intracytoplasmic sperm injection (ICSI), some patients present very low or even zero fertilization rates after ICSI. Artificial oocyte activation has been proposed as a suitable means to overcome this problem. We applied artificial oocyte activation in patients which presented a history either no fertilization, fertilization between 0 and 30% or fertilization between 30 and 50% in initial ICSI cycles. In the following treatment cycles, oocytes were activated after ICSI using a calcium ionophore. Fertilization, pregnancy and take-home baby rates were compared to the previous cycle without activation. For the groups with previously 0% or 1–29% fertilization, we noted higher fertilization rates and clinical pregnancy rates per embryo transfer. For the group with moderate fertilization, only fertilization rates differed but all other parameters were not significantly different. From these data we conclude that artificial oocyte activation has a great potential especially in patients which show a compromised fertilization rate below 30% in a standard ICSI cycle.

Synchronization between embryo development and endometrium is a contributing factor for rescue ICSI outcome

2012-02-10

Abstract: Recent evidence shows that the outcome of rescue intracytoplasmic sperm injection (ICSI) is unsatisfactory on account of a poor clinical pregnancy rate. These outcomes may be due to either the in-vitro ageing of cultured oocytes before ICSI or the asynchrony between the embryo developmental stage and the endometrial secretory pattern. To address the latter issue, this study performed a retrospective analysis of 534 fresh cycles after rescue ICSI and 64 frozen–thawed cycles in subsequent treatment. Rescue ICSI cycles were divided into three groups: group I included 469 fresh embryo-transfer (FET) cycles; group II included 74 FET cycles in which supernumerary good-quality embryos were also cryopreserved; and group III included 64 frozen–thawed transfer cycles. Group III was considered to have achieved better synchronization than group II. As a result, significantly higher clinical pregnancy (29.69%, 19/64 versus 10.81%, 8/74) and implantation (13.33%, 22/165 versus 5.13%, 8/156) rates were achieved in group III compared with group II (both P<0.05). Therefore, synchronization of embryo development with the endometrium is considered a contributing factor for rescue ICSI outcome. It is recommended that embryos derived from rescue ICSI cycles should be cryopreserved and subsequently used in frozen–thawed cycles.Intracytoplasmic sperm injection (ICSI) of unfertilized 1-day-old oocytes, called rescue ICSI, has frequently been performed in some infertility centres, when fertilization failure sometimes occurs in conventional IVF cycles. Recent studies showed that the outcome of rescue ICSI was unsatisfactory due to poor clinical pregnancy rates. One reason could be asynchrony between the embryo developmental stage and the endometrial secretory pattern. To address this issue, we performed a retrospective analysis of 534 fresh cycles after rescue ICSI (from January 2006 to January 2011) and 64 frozen–thawed transfer cycles in subsequent treatment (from January 2006 to May 2011) in our infertility centre. In this study, rescue ICSI cycles were divided into three groups. As there was no significant difference in women’s age (31.22±3.38 versus 31.11±3.27years) between groups II and III, we principally compared these two groups. Group II included 74 fresh embryo transfer cycles, in which supernumerary good-quality embryos were cryopreserved, and group III included 64 frozen–thawed transfer cycles. Group III was considered to have better synchronization than group II. As a result, significantly higher clinical pregnancy (29.69% versus 10.81%) and implantation (13.33% versus 5.13%) rates were achieved in group III compared with group II. Therefore, endometrial synchronization is considered a contributing factor for rescue ICSI outcome and embryos derived from rescue ICSI cycles should be cryopreserved and subsequently used in frozen–thawed cycles.

Anti-Müllerian hormone for the assessment of ovarian response in GnRH-antagonist-treated oocyte donors

2012-02-06

Abstract: Evidence regarding the role of anti-Müllerian hormone (AMH) among oocyte donors is limited and only involves gonadotrophin-releasing hormone (GnRH)-agonist-treated donors. This trial assessed the predictive ability of AMH for ovarian response among 108 oocyte donors treated with an antagonist protocol. In multivariate linear regression analysis, both AMH and age were independently associated with ovarian response (unstandardized coefficients 0.904 and −0.378, respectively). In receiver operating characteristic curve analysis, AMH performed better than age, but was a modest predictive marker for low (⩽6 oocytes) and excessive (>20 oocytes) ovarian response (area under the curve (AUC) 0.643 and 0.695, respectively). Similarly, a multivariate logistic model including AMH and age was also modest (AUC 0.651 and 0.697 for low and excessive responders, respectively). The predictive ability of AMH did not significantly alter when different thresholds were adopted, such as <4 oocytes for low response and >25 for excessive response (AUC 0.759 and 0.724, respectively). Among oocyte donors treated with a GnRH-antagonist protocol, although AMH was correlated with the number of oocytes retrieved, it demonstrates a modest ability in discriminating women with low or excessive ovarian response.Selection of oocyte donors is of paramount importance for the proper and more cost-efficient functionality of the oocyte donation programme. Despite the extensive literature regarding the efficacy of anti-Müllerian hormone (AMH) for predicting ovarian response among infertile patients, available evidence regarding the role of AMH in oocyte donors is considerably limited and involves only agonist down-regulated cycles. In this trial we assessed whether AMH can be considered a predictive marker for ovarian response among oocyte donors treated with a gonadotrophin-releasing hormone (GnRH)-antagonist protocol. According to our results, among oocyte donors treated with a GnRH-antagonist protocol, although AMH was correlated with the number of oocytes retrieved, the correlation is not strong and it appears that AMH has a modest predictive ability to discriminate women who are likely to demonstrate either impaired or excessive response to ovarian stimulation.

Anti-Müllerian hormone is highly expressed and secreted from cumulus granulosa cells of stimulated preovulatory immature and atretic oocytes

2012-02-02

Abstract: This study investigated anti-Müllerian hormone (AMH) expression and secretion from cumulus granulosa cells (GC) and steroidogenesis in follicular fluids (FF) with relation to oocyte maturational stages and fertilization capacity in large preovulatory follicles. This prospective study included 53 ovulatory women undergoing intracytoplasmic sperm injection. FF and cumulus GC from 140 large preovulatory follicles were individually obtained during oocyte retrieval. Main outcome measures were oocyte maturation, fertilization and embryo quality. FF were assayed for AMH, progesterone, 17β-oestradiol and testosterone. Cumulus GC were assayed for AMH mRNA expression. AMH mRNA expression and secretion in cumulus GC in preovulatory follicles containing germinal-vesicle (GV) and metaphase-I (MI) oocytes were significantly higher than follicles containing MII oocytes (P<0.01 and P<0.0001, respectively). In addition, FF AMH concentrations from atretic oocytes were significantly higher than from MII oocytes. No correlation was found between AMH expression and secretion to fertilization or embryo quality. FF of MI and GV oocytes had higher concentrations of testosterone and lower progesterone/oestradiol ratios than MII oocytes, and FF of atretic oocytes contained higher testosterone concentrations than FF of MII oocytes. AMH is highly expressed in and secreted from cumulus GC of preovulatory follicles containing premature and atretic oocytes.Anti-Müllerian hormone (AMH) is produced in the female exclusively by granulosa cells. AMH has recently been shown to be one of the most important markers of ovarian reserve and it is highly associated with ovarian follicular development. This study investigates AMH expression and secretion from cumulus granulosa cells (GC) and steroidogenesis in the follicular fluids (FF) with relation to oocyte maturational stages, and fertilization capacity in large preovulatory follicles. We conducted a prospective study with 53 ovulatory women undergoing intracytoplasmic sperm injection. FF and cumulus GC from 140 large preovulatory follicles were individually obtained during oocyte retrieval. The main outcome measures were oocyte maturation, fertilization and embryo quality. FF were assayed for AMH, progesterone, 17β-oestradiol and testosterone. Cumulus GC were assayed for AMH mRNA expression. AMH mRNA expression in cumulus GC and AMH concentrations in FF of preovulatory follicles containing premature oocytes (germinal vesicle (GV) and metaphase I (MI)) were significantly higher than preovulatory follicles containing mature oocytes (MII oocytes). In addition, FF AMH concentrations of atretic oocytes were significantly higher than FF AMH of MII oocytes. No correlation was found between AMH expression and secretion for fertilization or embryo quality. FF of preovulatory MI and GV oocytes had higher levels of testosterone and lower progesterone/oestradiol ratios than MII oocytes, and FF of atretic oocytes contained higher testosterone levels than FF of MII oocytes. This study shows that AMH is highly expressed in and secreted from cumulus GC of preovulatory follicles containing premature and atretic oocytes.

Successful pregnancy outcome following gamete intra-Fallopian transfer in a patient with Müllerian dysgenesis

Colin S.S. Lee, Aldrin T.M. Lie — 2012-02-02

Abstract: A 29-year-old lady with Müllerian dysgenesis was keen to have a baby. Clinically, she was medium built with well-developed secondary female sexual characteristics. There was a short and blind vagina. She had undergone surgery for an imperforated hymen. Her FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent gamete intra-Fallopian transfer (GIFT). Oocyte retrieval was carried out laparoscopically and a total of nine oocytes were retrieved. Four of the oocytes were transferred together with motile spermatozoa into the right Fallopian tube and the remaining five oocytes were inseminated with spermatozoa for IVF. Three embryos resulted and were frozen. She subsequently developed moderate ovarian hyperstimulation syndrome. Serum β-human chorionic gonadotrophin concentration 14days after GIFT was 1612IU/l. Her antenatal care was relatively uneventful until 31weeks of gestation when she was diagnosed to have intrauterine growth retardation and oligohydramnios. She then underwent an emergency Caesarean section at 32weeks of pregnancy delivering a normal baby.This case study describes a successful pregnancy outcome following gamete intra-Fallopian transfer (GIFT) in a woman with malformation of the vagina (Müllerian dysgenesis). A 29-year-old lady with Müllerian dysgenesis diagnosed at 16 years of age was keen to become pregnant. Upon examination, a decision was made for a William’s vulvovaginoplasty but as the patient was indecisive the surgery was deferred. Clinically, she is a medium-built lady with well-developed secondary female sexual characteristics. There was a short and blind vagina. Her serum FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent GIFT. Nine oocytes were retrieved through laparoscopy. Four of the oocytes were transferred together with motile sperm into the right Fallopian tube and the remaining five oocytes were inseminated with sperm for IVF. Three embryos resulted and were frozen. Serum β human chorionic gonadotrophin concentration measured 14 days after GIFT was 1612IU/l. An abdominal ultrasonography performed at 5 weeks showed one intrauterine gestational sac. Her antenatal care was uneventful until 31 weeks of gestation when she developed a deficiency of amniotic fluid in the amniotic sac. She then underwent an emergency Caesarean section at 32 weeks of pregnancy. She delivered a healthy, normal 1.24kg baby boy. Her post-natal care was uneventful.

Expression and localization of opioid receptors during the maturation of human oocytes

2012-02-24

Abstract: The endogenous opioid system has been characterized in some female reproductive organs, but little is known about the expression of these receptors in human oocytes. This study investigated the presence and differential distribution of the opioid receptors during the maturation of human oocytes. A total of 821 human oocytes from an intracytoplasmic sperm injection (ICSI) programme were studied including 213 at germinal-vesicle (GV) stage and 164 at metaphase-I (MI) stage and 444 failed fertilization metaphase-II (MII) oocytes. Additionally 31 MII oocytes corresponding to cases where ICSI was not attempted and 50 failed fertilization MII oocytes from the IVF programme were included. Western blot analysis revealed the presence of the delta (OPRD1), kappa (OPRK1) and mu (OPRM1) opioid receptors in human oocytes. The OPRK1 and OPRM1 immunostaining patterns changed during the maturation of the oocyte, while the OPRD1 pattern was the same throughout. In particular, OPRD1 were detected in peripheral tissue from the GV to the MII stage. OPRK1 were found peripherally at the GV stage, more internally at MI and homogeneously at MII. Finally, OPRM1 were located peripherally at the GV stage and homogeneously in MI and MII oocytes. Opioids may have a role in oocyte maturation, acting via receptors.The opioid system has been well characterized in the central nervous system, but it is now known that opioids also act in reproductive organs. However, little is known about the presence and function of this system in human oocytes and its role in their maturation. In this study, we investigated the presence and differential distribution of three (delta, kappa and mu) opioid receptors (proteins which bind the opioids) during the maturation of human oocytes. A total of 821 human oocytes (from 253 patients) not suitable for intracytoplasmic sperm injection (ICSI) or which did not develop into an embryo after ICSI were studied. Thus, we have verified the presence of the delta, kappa and mu opioid receptors in human oocytes. The kappa and mu localization changed during the maturation of the oocyte, while the Delta localization was the same throughout. In particular, the delta receptor was detected in the periphery of the oocyte. On the other hand, the kappa receptor was found peripherally at the beginning, more internally during maturation and homogeneously at the end of maturation. Finally, the Mu receptor was located peripherally at the beginning of maturation and homogeneously in the rest of the maturation stages. This finding suggests a possible role for opioids, acting via receptors, in the maturation of the oocyte.

Taurine attenuates maternal and embryonic oxidative stress in a streptozotocin-diabetic rat model

Mahesh Mysore Shivananjappa, Muralidhara — 2012-01-30

Abstract: Oxidative stress mechanisms have been implicated in congenital anomalies and morbidity/mortality of fetus/newborn in diabetic pregnancy. Numerous antioxidant treatments have shown varied beneficial effects in improving both maternal and fetal outcomes. The present study examined the propensity of taurine to attenuate the degree of embryopathy and oxidative stress among pregnant diabetic rats. Adult rats (CFT-Wistar) were rendered diabetic with an acute dose of streptozotocin (STZ; 45mg/kg bodyweight) on gestation day (GD) 4. Both Diabetic and non-diabetic dams were given oral supplements of taurine (0.5 and 1g/kg bodyweight/day) from GD 5 to GD 12. Maternal diet intake, bodyweight gain and urine output were monitored and dams were killed on GD 13. Markers of oxidative stress were determined in embryos and maternal livers. STZ treatment induced marked embryopathy (32%) and taurine supplements markedly reduced the degree of embryopathy (54% protection). The STZ-induced higher oxidative stress was significantly attenuated in rats given taurine supplements (P<0.05) and a similar effect was seen in embryos (P<0.05). These data suggest that dietary taurine during pregnancy provides significant protection against diabetes-induced oxidative stress in both the mother and the embryos and thus may serve as a therapeutic supplement during diabetic pregnancy.Diabetes during pregnancy affects >5% of all pregnancies, causing reproductive abnormalities that enhance spontaneous abortion – congenital anomalies, morbidity and mortality of both mother and fetus/newborn. One of the major mechanisms is increased oxidative stress caused by hyperglycaemia and the most prominent anti-teratogenic effect was achieved using antioxidative agents. Management of oxidative stress is considered, along with tight glycaemic control, to be beneficial both before conception and during pregnancy. Taurine, a ubiquitous amino acid found in almost all mammalian tissues, constitutes more than 50% of free amino acids. The aim of the study was to determine whether oral taurine supplementation given to pregnant diabetic rats during the post-implantation period could reduce embryo lethality and protect the developing embryos against maternal hyperglycaemia-induced oxidative stress. Adult rats were rendered diabetic with an acute dose of streptozotocin on gestation day (GD) 4. Both diabetic and non-diabetic dams were administered oral taurine for a period of 8days (GD 5–13). Maternal diet intake, bodyweight gain and urine output were monitored and dams were killed on GD 13. Markers of oxidative stress and antioxidant defences were studied in embryos and maternal livers. STZ induced marked embryopathy (32%) and taurine supplementation offered significant protection (54%). Taurine significantly offset diabetes-associated oxidative stress in the embryos of diabetic rats. These data suggest that dietary taurine supplementation during pregnancy provides significant protection against diabetes-induced oxidative stress both in mother and embryos and thus may serve as a therapeutic supplement under diabetic pregnancy.

Matrix metalloproteinases 1, 2, 3 and 9 functional single-nucleotide polymorphisms in idiopathic recurrent spontaneous abortion

Nina Pereza, Saša Ostojić, Marija Volk, Miljenko Kapović, Borut Peterlin — 2012-01-27

Abstract: Idiopathic recurrent spontaneous abortion (IRSA) has been associated with abnormalities in the remodelling of endometrial extracellular matrix, as well as aberrant matrix metalloproteinase (MMP) gene expression in endometrium of IRSA women and chorionic villi of IRSA concepti. This study investigated the association of five functional MMP gene promoter polymorphisms (MMP1 −1607 1G/2G, MMP2 −735 C/T, MMP2 −1306 C/T, MMP3 −1612 5A/6A and MMP9 −1562 C/T) with IRSA. A total of 149 couples with at least three consecutive IRSA and 149 fertile couples were included in a case–control study. Genotype analysis was performed using PCR restriction fragment length polymorphism. Statistically significant differences were found in distributions of MMP2 −735 CT (chi-squared 10.21, P=0.006; OR 2.15, 95% CI 1.34–3.45, P=0.001), and MMP9 −1562 CC (chi-squared 9.06, P=0.010; OR 2.21, 95% CI 1.30–3.80, P=0.004) between IRSA women and controls. Combined analysis of MMP gene polymorphisms did not increase their predictive value. There were no statistically significant differences in genotype and allele frequencies of any polymorphism between IRSA men and controls. MMP2 −735 C/T and MMP9 −1562 C/T functional gene polymorphisms might be associated with an increased risk of IRSA in women.Considering the insufficient knowledge on genetic contribution to pregnancy loss, studies on genetic causes of idiopathic recurrent spontaneous abortion (IRSA) are of great importance. Development of a histologically and functionally normal endometrium is critical for subsequent endometrial decidualization, receptivity and implantation. The proper communication and interaction between maternal decidual cells and the embryo is essential for the establishment of a functional fetal–maternal interface. IRSA has been associated with abnormalities in the remodelling of endometrial extracellular matrix, as well as aberrant matrix metalloproteinase (MMP) gene expression in endometrium of IRSA women and chorionic villi of IRSA concepti. The aim of this study was to investigate the association of five functional MMP gene promoter polymorphisms with IRSA. A total of 149 couples with at least three consecutive IRSA and 149 fertile couples were included in a case–control study. Genotype analysis was performed using polymerase chain reaction and restriction fragment length polymorphism. Statistically significant differences were found in distribution of MMP2 −735 CT and MMP9 −1562 CC genotypes between IRSA and control women. Combined analysis of MMP gene polymorphisms did not increase their predictive value. There were no statistically significant differences in distribution of genotype and allele frequencies of any polymorphism between IRSA men and controls. Our results demonstrate that MMP2 −735 C/T and MMP9 −1562 C/T functional gene polymorphisms might be associated with an increased risk of IRSA in women.

No clinical relevance of the height of fundal indentation in subseptate or arcuate uterus: a prospective study

2012-02-06

Abstract: The American Fertility Society has classified the arcuate uterus as a minor malformation with a benign clinical behaviour. The aim of this prospective study was to verify whether there is any scientific basis for this differentiation. Patients with at least one early miscarriage and a subseptate or arcuate uterus were admitted for hysteroscopic metroplasty. Patients were allocated to a subseptate uterus group, with an indentation of 1.5cm or more, or an arcuate uterus group, with a smaller indentation. The miscarriage rates after metroplasty were similar between the two groups (14.0% in the subseptate uterus group versus 11.1% in the arcuate uterus group). Before metroplasty, the miscarriage rates were significantly higher in subseptate uterus group, as well as in the arcuate uterus group (both P<0.001). According to these results, there is no evidence to support that the arcuate uterus has a different effect on the reproductive outcome in comparison to the subseptate uterus, neither before nor after surgical correction of the anomaly. Since there is no scientific basis for a separate classification of the arcuate uterus, a review of the classifications of uterine congenital anomalies should be considered as necessary.Congenital uterine malformations have been classified by the American Fertility Society (AFS) since 1988. Although the AFS classification received wide acceptance and is still the most broadly used system, it is associated with various limitations in effective categorization of the anomalies. It is interesting that, until now, none of the other available options have been able to effectively replace the AFS system. Numerous papers indicate septate or subseptate (partial septate) uterus (AFS class V) is a possible cause of an unfavourable pregnancy outcome. Arcuate uterus (AFS class VI), a slight malformation similar to septate uterus, should differ from septate or subseptate uterus, because this ‘minor’ malformation should behave benignly with respect to the septate uterus. The aim of this study was to scientifically validate the difference between the arcuate and subseptate uterus in their effect on reproductive outcome through the results of a metroplasty in both groups of patients. A group of 96 patients, who underwent metroplasty after at least one early miscarriage, was divided into two groups according to the severity of the congenital uterine malformation. Our results indicate that there are no differences in pregnancy outcome after metroplasty either in patients with septate or arcuate utera. The poor pregnancy outcome in women with septate uterus seems not to be correlated to the dimension of the septum itself. There are no scientific bases for a separate classification of the arcuate uterus and it is proposed that a review of the classification of uterine congenital anomalies is necessary.

Hellenic National Authority of Medically Assisted Reproduction

Haris E. Cazlaris, Xeni Skorini-Paparrigopoulou, Basil C. Tarlatzis — 2012-02-02

The recent article by contains a serious inaccuracy regarding the Hellenic National Authority of Medically Assisted Reproduction: it is incorrect that it has “not yet functioned”.

Effect of adenomyosis on implantation

J.M. Vila-Vives, J.A. Martínez-Conejero, A. Pellicer — 2012-02-20

We read with great interest the excellent review on adenomyosis and infertility by . Despite its considerable clinical relevance, there are two points that we would like to elaborate, namely the relationship between adenomyosis and infertility, and its possible mechanism of action.

Response: Effect of adenomyosis on implantation

V. Campo, S. Campo, G. Benagiano — 2012-02-29

We thank for highlighting their clinical results, which provide, for the first time, direct evidence that implantation may not be affected in women with adenomyosis. It is unfortunate that their paper became available after we had completed the revision of our review (), because their clinical data contradict the metabolic and molecular information previously available. New information becomes constantly available and it is inevitable that a cut-off date must be applied to any paper in press. Ours was 31 July 2011.

Adenomyosis and infertility

L.S. Louis, S. Saso, J. Chatterjee, E. Barsoum, M. Al-Samarrai — 2012-02-27

We read with interest the review by , which undoubtedly has enhanced the medical knowledge on this subject, particularly with regard to the emerging field of fertility-sparing cytoreductive surgery for large adenomyomata. However, the article fails to discuss the role of IVF following such surgery, although the paper by , which touches on this subject, is mentioned. Previous papers () have reported successful live births following cytoreductive surgery. However, in these reports all the patients conceived spontaneously following surgery.

Response: Adenomyosis and infertility

S. Campo, V. Campo, G. Benagiano — 2012-02-27

The letter from mentions that in our review () we failed to discuss the role of IVF following cytoreductive surgery. The fact of the matter is that data on treatment of adenomyosis-associated infertility are scarce and largely based on single cases or small series. Furthermore, on the specific issue of the role of IVF following conservative surgery, no conclusion can be drawn because of lack of information. The paper by states: “Twenty-six women (25.0%) wished to conceive following the surgical removal of the adenomyosis. … Of these, four women conceived spontaneously and 12 women conceived by IVF/ET. Two women who had IVF/ET experienced a spontaneous abortion (at 5weeks and 16weeks); 14 went to term”. We do not believe that this information sheds any light on whether IVF should be preferred following cytoreductive surgery. Therefore, there is no valid answer to the question raised by Louis et al., namely how many of them attempted IVF beforehand. We therefore agree that, as of today, we cannot “draw a reliable conclusion regarding the success of IVF treatment following cytoreductive surgery and counsel patients accordingly”.