Maternal body mass index is not associated with assisted reproductive technology outcomes.

The effects of body mass index (BMI) on assisted reproductive technology (ART) outcomes such as ovarian dysfunction, poor ovum quality, and endometrial dysfunction have been studied; however, many aspects remain controversial. Therefore, we retrospectively investigated the relationship between BMI and ART outcomes. For 14,605 oocyte retrieval cycles at our hospital between January 2016 and December 2020, BMI was divided into five groups (< 18.5, 18.5-20.0, 20.0-22.5, 22.5-25.0, ≥ 25 kg/m2) and measured before oocyte retrieval. The normal fertilization and high-grade blastocyst rates were compared. In addition, in the 7,122 frozen-thawed embryo transfers (FET) with highest-grade embryos, the clinical pregnancy, miscarriage, and live birth rates were investigated in the five BMI groups. Multiple regression analysis on normal fertilization and high-grade blastocyst rates revealed no statistically significant differences. Furthermore, after propensity score matching on FET, there was no significant difference in clinical pregnancy, miscarriage, and live birth rates in the BMI groups. BMI is a risk factor for complications during pregnancy; however, it does not affect ART outcomes. Therefore, we believe weight guidance should be provided to women with obesity at the start of infertility treatment, but treatment should not be delayed.

Exome-based genome-wide screening of rare variants associated with the risk of polycystic ovary syndrome.

Purpose: Genetic factors associated with the risk of polycystic ovary syndrome (PCOS) remain largely unknown. Here, we conducted an optimal sequence kernel association test (SKAT-O), an exome-based rare variant association study, to clarify whether rare variants in specific genes contribute to the development of PCOS. Methods: SKAT-O was performed using exome data of 44 Japanese patients with PCOS and 301 control women. We analyzed frequencies of rare probably damaging variants in the genome. Results: Rare variants of GSTO2 were more commonly identified in the patient group than in the control group (6/44 vs. 1/301; Bonferroni-corrected p-value, 0.028), while the frequencies of variants in other genes were comparable between the two groups. The identified GSTO2 variants were predicted to affect the function, structure, stability, hydrophobicity, and/or the formation of intrinsically disordered regions of the protein. GSTO2 encodes a glutathione transferase that mediates the oxidative stress response and arsenic metabolism. Previously, common variants in GSTO2 and its paralog GSTO1 were associated with the risk of PCOS. Conclusions: The results indicate that there are no genes whose rare variants account for a large fraction of the etiology of PCOS, although rare damaging variants in GSTO2 may constitute a risk factor in some cases.

Impact of paternal age on IVF and pregnancy outcomes with only normal sperm parameters.

OBJECTIVE: Previous reports on advanced paternal age effects on assisted reproductive technology (ART) vary considerably and those on frozen-thawed embryo transfer (FET) are rare. We investigated whether paternal age affects in vitro fertilisation (IVF) and FET pregnancy outcomes. MATERIALS AND METHODS: 1657 IVF cycles performed from January 2014 to May 2018 were retrospectively investigated excluding cases of poor semen parameters. Paternal and maternal ages were categorised into groups, namely, <35, 35-39 and ≥ 40 years, to compare normal fertilisation (2 PN (pronuclei)) and high-quality blastocyst rates. Furthermore, 741 FET cycles were investigated and pregnancy, live birth and miscarriage rates were compared. RESULTS: For the maternal age group (35-39), the 2 PN rate was significantly higher with paternal age group of <35 than groups of 35-39 and ≥ 40 (median%, <35 vs. 35-39 vs. ≥40 = 100.0 vs. 71.4 vs. 77.7; P = 0.005). The miscarriage rate was significantly higher with paternal age group of ≥40 than that of <35 and 35-39 when maternal age was <35 (median %, <35 vs. 35-39 vs. ≥40 = 13.1 vs. 7.8 vs. 33.3; P = 0.038). CONCLUSION: Our findings show that when maternal age was <35, advanced paternal age reduces the normal fertilisation rate and increases the FET miscarriage rate when maternal age was 35-39.

Aging increases oxidative stress in semen.

PURPOSE: As age increases, oxidative stress increases, sperm motility decreases, and DNA fragmentation increases. To date, reports of age-related effects on semen have focused on reactive oxygen species (ROS) or total antioxidant capacity (TAC) as indicators of oxidative stress. However, assessments of ROS and TAC must be considered within a more comprehensive context in order to correctly evaluate oxidative stress and interpret findings. In this regard, the purpose of this study was to investigate the relationship between the static oxidation reduction potential (sORP) and paternal age with the goal of using the sORP as an indicator of semen oxidative stress. MATERIALS AND METHODS: Semen samples from 173 men were analyzed for the following parameters: volume, motility, and beat cross frequency (BCF). The sORP was measured by using the MiOXSYS™ system. The correlation between semen parameters and the sORP level was analyzed as a function of age. The rate of sORP positivity was compared between men <34 and ≥34 years of age, with a positive sORP defined as a level ≥1.38. RESULTS: Volume, motility, and BCF were negatively correlated with age (p<0.001). The semen sORP level was positively correlated with age (p<0.05). The rate of sORP positivity was significantly increased in men ≥34 years of age compared with that in men <34 years of age (33% compared with 12%, respectively; p<0.01). CONCLUSIONS: The sORP may play a role in age-related decreases in semen parameters (volume, motility, and BCF). The rate of sORP positivity increased significantly after 34 years of age.

Circulating steroids and mood disorders in patients with polycystic ovary syndrome.

Aberrant androgen metabolism is a characteristic feature of polycystic ovary syndrome (PCOS). Various androgens as well as their precursors and metabolites can accumulate in the blood of PCOS patients. Although these steroids include neuroactive steroids, such as allopregnanolone and androstenedione (Δ4A), it remains unknown whether altered blood steroid levels contribute to the high risk of mood disorders in PCOS. In this study, we measured blood levels of 11 steroids in 25 PCOS patients using liquid chromatography-tandem mass spectrometry and chemiluminescent enzyme immunoassay, and assessed the psychological status of these patients using the Hospital Anxiety and Depression Scale (HADS) questionnaire. We also examined age and the degree of metabolic abnormalities of each patient. Steroid values of the patients were compared to our previous data from 31 eumenorrheic women. As a result, 20 patients exhibited aberrant blood levels of one or more of the 11 tested steroids. In most cases, Δ4A and allopregnanolone levels were within or close to the reference ranges. Levels of four steroids were negatively correlated with patients' age, while no correlation was observed between steroid values and metabolic conditions. Seven patients showed high HADS scores. HADS scores were correlated with blood Δ4A levels even after stratifying by body mass indexes, but not with the levels of other steroids or clinical data. These results indicate that the high frequency of anxiety and depression in PCOS patients cannot be ascribed to altered blood levels of a specific steroid, although there may be a weak association between circulating Δ4A levels and psychological conditions of the patients.

Clinical outcomes of microdissection testicular sperm extraction and intracytoplasmic sperm injection in Japanese men with Y chromosome microdeletions.

PURPOSE: We investigated the clinical results of Japanese men with Y chromosome microdeletions. METHODS: This study retrospectively examined 2163 azoospermic or severe oligozoospermic patients. We investigated the frequency of azoospermia factor (AZF) deletions and sperm retrieval rate (SRR) by microTESE in patients with these deletions, then analyzed the ICSI outcomes. RESULTS: Azoospermia factor deletions were found in 201 patients. SRR was significantly higher than that of the control group (74.0% vs 20.4%, P < .001). Thirty-three couples underwent ICSI using testicular spermatozoa retrieved by microTESE, and eight couples underwent ICSI using ejaculatory spermatozoa. The fertilization rate and clinical pregnancy rate per embryo transfer cycle were significantly higher in the ejaculatory group than that of the testicular group (66.4% vs 43.7%, P < .001, 53.3% vs 24.7%, P = .03, respectively). When compared with the control group, the fertilization rate was significantly lower in the testicular group with AZFc microdeletions (43.7% vs 53.6%, P < .001). CONCLUSIONS: Our study highlights that although microTESE in azoospermic men with AZFc microdeletions led to a higher SRR, ICSI outcomes of these men were worse than that of men without AZF deletions, even if testicular spermatozoa were retrieved.

Analysis of 122 triplet and one quadruplet pregnancies after single embryo transfer in Japan.

RESEARCH QUESTION: What is the prevalence of triplet and quadruplet pregnancies after single embryo transfer (SET) in Japan. DESIGN: A retrospective observational study was conducted on 274,605 pregnancies after 937,848 SET cycles in registered assisted reproductive technology (ART) data from the Japanese ART national registry database between 2007 and 2014. A questionnaire survey of ART centres was also conducted. Data on pregnancies with embryo division into three or more after SET were analysed. RESULTS: According to the Japanese ART national registry database, SET resulted in 109 triplet pregnancies (0.04% of pregnancies), and the questionnaire reports from 31 centres revealed 33 triplet and one quadruplet pregnancies. After exclusion of 20 duplicated cases, 122 triplet and one quadruplet pregnancies included 46 monochorionic (one gestational sac [37.4%]), 18 dichorionic (two gestational sacs [14.6%]) and 59 trichorionic pregnancies (three gestational sacs [48.0%]). Compared with singleton pregnancies, patients with monozygotic triplet or quadruplet pregnancies were less frequently diagnosed with unexplained infertility (P = 0.004), more often received gonadotrophin injections for ovarian stimulation in 39 cases with information available (P = 0.021) and underwent more blastocyst transfers and assisted hatching (P = 0.002 and P < 0.001, respectively). The proportion of live birth, defined as at least one baby born, excluding induced abortion, was 64.6% (73/116 pregnancies) of monozygotic triplet or quadruplet pregnancies. CONCLUSIONS: Combined Japanese ART national registry and survey data revealed 122 triplet and one quadruplet pregnancies, the majority after cryopreserved embryo transfer. Most were conceived after blastocyst transfer and often after assisted hatching, which are potential risk factors for zygotic splitting.

Clinical outcomes of transfer of frozen and thawed single blastocysts derived from nonpronuclear and monopronuclear zygotes.

PURPOSE: In assisted reproductive technology, normal zygotes are bipronuclear (2PN) during fertilization confirmation; however, sometimes, nonpronuclear zygotes (0PN) and monopronuclear zygotes (1PN) are found during routine observations. METHODS: To elucidate the clinical usefulness of in vitro-fertilized embryos, we investigated the rates of clinical pregnancy, live birth, miscarriage, and congenital abnormality after transfer of frozen-thawed 1PN- and 0PN-derived single blastocysts at Denentoshi Ladies Clinic, Kanagawa, Japan. RESULTS: The rates of pregnancy and live birth for 1PN-derived blastocysts obtained by conventional in vitro fertilization were 37.5% and 27.1%, respectively, which was not significantly different from those for 2PN-derived blastocysts; however, the rates for 0PN-derived blastocysts were significantly lower. The pregnancy and live birth rates for 0PN-derived embryos obtained by intracytoplasmic sperm injection (ICSI) were 45.7% and 34.8%, respectively, which was not significantly different from those for 2PN-derived blastocysts; however, the rates for 1PN-derived blastocysts were significantly lower (4.0% for both) than those for 2PN- and 0PN-derived blastocysts. No congenital abnormalities were found in infants resulting from transfer of 0PN- or 1PN-derived blastocysts. CONCLUSIONS: Both 1PN- and 0PN-derived blastocysts can be used for embryo transfer; however, care should be taken in making decisions about 1PN-derived blastocysts, especially if they are obtained by ICSI.

Distinctive subpopulations of the intestinal microbiota are present in women with unexplained chronic anovulation.

RESEARCH QUESTION: Do gut microbiota associate with the ovulatory cycle in women showing normogonadotrophic anovulation? In humans, the gut microbiota affects diverse physiological functions and dysbiosis (microbial imbalance) may lead to pathological syndromes. However, there is comparatively little information on the relevance of gut microbiota to reproductive functions in women. Here, a group of women with idiopathic chronic anovulation were examined, who do not exhibit any apparent endocrinological disorder, as they are suitable for investigating the relationship between intestinal bacteria and ovulatory disorders. DESIGN: A prospective observational cohort study was performed on two groups of women who did not exhibit apparent endocrinological disorders but showed either irregular menstrual cycles (IMC group) or normal menstrual cycles (controls). The bacterial composition of faeces from rectal swabs from the women was analysed using next-generation sequencing based on bacterial 16SrRNA genes. RESULTS: A metagenomic analysis indicated that the two groups of women had significant differences in 28 bacterial taxa in their faeces. Prevotella-enriched microbiomes were more abundant in the IMC group, whereas Clostridiales, Ruminococcus and Lachnospiraceae (butyrate-producing bacteria) were present at lower levels in the IMC group. CONCLUSIONS: Distinctive subpopulations of intestinal microbiota were identified in women with unexplained chronic anovulation. The results indicate that gut microbiota could be associated with ovarian functions.

Parental age and gene expression profiles in individual human blastocysts.

The epigenetic status of the genome changes dynamically from fertilization to implantation. In addition, the physiological environment during the process of gametogenesis, including parental age, may affect the epigenome of the embryo after fertilization. It is important to clarify the influence of parental age on gene expression in the embryo in terms of transgenerational epigenetics to improve the techniques currently used in assisted reproductive medicine. Here, we performed single-embryo RNA-seq analysis on human blastocysts fertilized by intracytoplasmic sperm injection, including from relatively elderly mothers, to elucidate the effects of parental age on the embryonic gene expression profile. We identified a number of genes in which the expression levels were decreased with increasing maternal age. Among these genes, several are considered to be important for meiotic chromosomal segregation, such as PTTG1, AURKC, SMC1B and MEIKIN. Furthermore, the expression levels of certain genes critical for autophagy and embryonic growth, specifically GABARAPL1 and GABARAPL3, were negatively correlated with advanced paternal age. In addition, levels of transcripts derived from major satellite repeats also decreased as the maternal age increased. These results suggest that epigenetic modifications of the oocyte genome may change with parental age and be transmitted to the next generation.

The significance of serum anti-Müllerian hormone (AMH) levels in patients over age 40 in first IVF treatment.

PURPOSE: Although studies of serum anti-Müllerian hormone (AMH) in predicting ovarian reserve are numerous, many studies utilized patients under age 40. However, the assessment of ovarian reserve is especially critical in older infertile women. This study evaluates the significance of AMH level in patients over age 40 at the time of their first in vitro fertilization (IVF) treatment. METHODS: Forty-nine women over age 40 were studied. Although serum samples were taken prior to their IVF treatments, the data of serum AMH of patients were not taken into consideration to determine the therapy strategy, including follicle induction in which clomiphene citrate and human menopausal gonadotropin were used. RESULT(S): Twelve out of 49 patients achieved a clinical pregnancy (24.4 %). There was a positive correlation between serum AMH levels and the number of oocytes retrieved (P < 0.0001). The ROC curve analysis for prediction of poor ovarian response, ≤3 retrieved oocytes, showed that the optimum cut-off level was < 1.0 ng/mL for AMH. The lower AMH group (AMH < 1.0 ng/ml) showed less chance of undergoing embryo transfer than the higher AMH group (AMH ≥1.0 ng/ml). There was no difference in pregnancy rate between the two groups. Five out of 12 pregnant women exhibited AMH levels of less than 0.4 ng/ml. CONCLUSION(S): Assessment of serum AMH concentration in older patients is useful for the prediction of oocytes numbers which may be obtained in IVF. A cut-off level of 1.0 ng/ml AMH can be used to predict poor ovarian response. This cut-off level of AMH of 1.0 ng/ml might be useful to predict whether patients could have an embryo transfer, but had no power to predict achieving pregnancy. On the other hand, our data also showed that patients over age 40 with extreme low levels of AMH still had a chance of pregnancy.

Clinical outcome of frozen blastocyst transfer; single vs. double transfer.

PURPOSE: Frozen embryo transfer has been established as an indispensable ART procedure for both the effective use of surplus embryos and the prevention of ovarian hyperstimulation syndrome. The frequency of frozen embryo transfer is increasing in our clinic, and we report that frozen embryo transfer is effective for patients with repeat failures. We present our clinical outcome of frozen blastocyst transfer (FBT). METHODS: In 2006, 470 patients received FBT (562 cycles (IVF: 354 cycles; ICSI: 208 cycles)). One frozen blastocyst was transferred in 412 cycles (335 patients) and two blastocysts were transferred in 150 cycles (135 patients). Assisted hatching was performed in all cases. RESULTS: In 412 cycles (average age: 34.6 years) who received a single FBT, the rate of clinical pregnancy per cycle was 40.7%, the live birth rate was 29.1%, the abortion rate was 21.6%, the ectopic pregnancy rate was 1.2%, the frequency of monochorionic twins was 2.3%, and the cesarean section rate was 38.3%. In 150 cycles (average patient age 34.8 years) who received two FBTs, the clinical pregnancy rate was 46%, the live birth rate was 35.3%, the abortion rate was 16.3%, the ectopic pregnancy rate was 4.4%, the frequency of twins was 15.9% and the cesarean section rate was 39.6%. A significant difference in the ectopic pregnancy rate and the twinning rate was found between single transfers and double transfers (P < 0.05). When IVF and ICSI were compared, there was no statistically significant difference in the abortion rate, the ectopic pregnancy rate, and the cesarean section rate. CONCLUSIONS: The clinical pregnancy rate was similar for the transfer of one and two blastocysts. Single FBT decreases obstetrical risk without reducing the pregnancy rate.

The decrease of serum luteinizing hormone level by a gonadotropin-releasing hormone antagonist following the mild IVF stimulation protocol for IVF and its clinical outcome.

PURPOSE: While performing the mild ovarian stimulation protocol with a GnRH antagonist, the pregnancy rate was compared between the groups, which were divided by the degree that the luteinizing hormone (LH) level decreased. MATERIALS AND METHODS: Patients aged 27 to 42years (36.1 +/- 3.79) underwent 308 IVF cycles who opted for IVF via the mild ovarian stimulation protocol began clomiphene citrate on day 3 and recombinant FSH on day 5. A GnRH antagonist was administered when the dominant follicle reached 14mm. Serum LH was measured at the time of GnRH antagonist administration and at the time of hCG injection. The pregnancy rate and implantation rate were compared between 50 cycles in which the LH level dropped less than one-third and the control (LH level within 1/3). RESULT(S): The pregnancy rate for the group in which the LH level fell less than one third was 18%. Conversely, the pregnancy rate for the control group was 39%. The implantation rate was 18% for the less than one-third group and 26% for the control group. Both the pregnancy rate and the implantation rate for the group in which the LH level fell less than one-third were significantly lower than that of control (p < 0.02). CONCLUSION(S): When performing the mild ovarian stimulation protocol, serum LH should be followed. If the serum LH level is less than one-third at the time of hCG injection, both the pregnancy rate and implantation rate are significantly lower.

Mild stimulation with clomiphene citrate in combination with recombinant follicle-stimulating hormone and gonadotropin-releasing hormone antagonist and its influence on serum estradiol level and pregnancy rate.

Aim: The mild ovarian stimulation protocol for in vitro fertilization (IVF) is carried out to minimize adverse side-effects as well as cost. While performing mild ovarian stimulation with a gonadotropin-releasing hormone (GnRH) antagonist, the pregnancy rate was examined in cases that exhibited a serum estradiol (E2) drop down. Methods: In this study, 174 patients who requested mild ovarian stimulation for IVF began clomiphene citrate on day 3 and recombinant follicle-stimulating hormone (FSH) on day 5 of their menstrual cycles. A GnRH antagonist was administered when the dominant follicle reached a diameter of 14 mm. Serum luteinizing hormone and estradiol were measured at the time of GnRH antagonist administration and at the time of human chorionic gonadotropin (hCG) injection. Pregnancy rates and implantation rates were compared between 24 cycles in which the E2 level fell at the time of hCG injection and 150 cycles in which it did not fall. Results: The pregnancy rate in the cases in which the E2 level fell (25% decrease) at the time of hCG injection was significantly lower than it was in the cases in which it did not fall (16.7 vs 41.0%). The implantation rate for the cases in which the E2 level fell was also lower than that of the control group (7.0 vs 31.0%). There was no significant difference in the number of good-quality embryos between the two groups. Conclusion: When performing the mild ovarian stimulation protocol, serum E2 should be followed. It is prudent to avoid embryo transfer in the same cycle in cases that exhibit E2 drop down. (Reprod Med Biol 2008; 7: 85-89).

Location of endometrial polyp and pregnancy rate in infertility patients.

OBJECTIVE: To assess endometrial polyp location and abnormal endometrial findings and their impact on pregnancy rate. DESIGN: Retrospective study. SETTING: Reproductive clinic in Kanagawa, Japan. PATIENT(S): A retrospective study was conducted on 230 infertility patients who had an endometrial polyp, as suspected on the basis of ultrasound and as diagnosed by hysteroscopy. INTERVENTION(S): Polyps were excised by either polypectomy or curettage. All samples were examined for pathology. The endometrial cavity was subdivided into five area categories: uterotubal junction, anterior uterine wall, posterior uterine wall, lateral uterine wall, and multiple. The patients received < or =6 months of follow-up; pregnancy rates were compared between the five subdivisions. MAIN OUTCOME MEASURE(S): Pregnancy rates. RESULT(S): The incidence of endometrial polyps was as follows, by location: uterotubal junction, 8.0%; posterior uterine wall, 32.0%; anterior uterine wall, 15.4%; lateral uterine wall, 9.2%; and multiple, 35.4%. The pregnancy rate after surgery was as follows, by location: uterotubal junction, 57.4%; posterior uterine wall, 28.5%; anterior uterine wall, 14.8%; lateral uterine wall, 18.8%; and multiple, 40.3%. Endometrial hyperplasia was found in 6.9% of the cases. The pregnancy rate after surgery at the uterotubal junction was significantly higher than that of other locations. CONCLUSION(S): Endometrial polyps are commonly found on the posterior wall of the uterus; however, excision of polyps that were located at the uterotubal junction significantly improved the pregnancy rate. Endometrial polyps should be categorized by both size and location.

Monozygotic multiple gestation following in vitro fertilization: analysis of seven cases from Japan.

We present a series of monozygous multiple gestations achieved following in vitro fertilization (IVF): one case of monochorionic triplet pregnancy and six cases of dizygotic triplet pregnancy. From September 2000 to December 2006, all patients achieving clinical pregnancy by ART were reviewed (n = 2433). A 37 year-old woman who delivered a healthy singleton after IVF returned two years later for FET, and a single blastocyst was transferred. This also resulted in pregnancy, but TV-USG revealed a single gestational sac with three distinct amniotic sacs, each containing a distinct fetal pole with cardiac activity. This pregnancy was electively terminated at nine weeks' gestation. An additional six cases of dizygotic triplets established after fresh embryo transfer (no ICSI or assisted hatching) are also described. Of these, one resulted in a miscarriage at eight weeks' gestation and five patients have an ongoing pregnancy. This case series suggests the incidence of dizygotic/monochorionic triplets following IVF is approximately 10 times higher than the expected rate in unassisted conceptions, and underscores the importance of a conservative approach to lower the number of embryos at transfer. The role of embryo transfer technique and in vitro culture media in the twinning process requires further study.

High concentrations of lactoferrin in the follicular fluid correlate with embryo quality during in vitro fertilization cycles.

OBJECTIVE: To investigate the level of lactoferrin (LF) in follicular fluid (FF), as well as the correlation between the LF concentration of FF, fertilization rate, and embryo quality. DESIGN: Retrospective study. SETTING: In vitro fertilization program at the Showa University School of Medicine, Tokyo, Japan. PATIENT(S): Follicular fluid was obtained from 35 IVF patients, aged 25-34 years. INTERVENTION(S): In total, 70 FF and serum samples were collected at the time of oocyte harvesting (44 of the oocytes were fertilized, and 26 were not). The LF concentration in FF was assayed via ELISA. RESULT(S): The mean LF concentration in FF of the fertilized group (500.2 +/- 35.5 ng/mL) was significantly higher than that in the unfertilized group (379.7 +/- 45.1 ng/mL). A positive correlation between follicular size and LF concentration was not found (r = 0.214). However, a high LF concentration in FF was indicative of good embryo quality. CONCLUSION(S): High LF concentrations in FF correlate with both a high fertilization rate and good embryo quality.

Clinical outcomes of two different endometrial preparation methods for cryopreserved-thawed embryo transfer in patients with a normal menstrual cycle.

Aim: To compare the clinical outcomes of cryopreserved-thawed embryo transfer among patients with a normal menstrual cycle who had natural or hormone-replacement cycles. Methods: From January 2004 to June 2006, cryopreserved embryos following conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) were thawed and transferred in a total of 720 natural cycles and 136 hormone-replacement cycles. Results: Cryopreserved-thawed embryo transfer in patients who had a natural or hormone-replacement cycle resulted in clinical pregnancy in 43.1% and 40.4%, respectively; a rate of miscarriage of 14.5% and 23.6%, respectively; and a rate of ongoing pregnancy and delivery of 36.5% and 30.9%, respectively. None of these differences were statistically significant. Conclusions: Patients with a normal menstrual cycle who have natural or hormone-replacement cycles can be expected to have comparable clinical outcomes with cryopreserved-thawed embryo transfer. (Reprod Med Biol 2007; 6: 53-57).

Serum steroids concentration might be used for monitoring the growth of follicles in friendly in vitro fertilization.

Background: Steroid levels have been used as the predictive parameters for oocyte maturation and embryo development. In the present study, estradiol and progesterone concentrations in the follicular fluid and serum were evaluated in conventional in vitro fertilization (IVF; follicle stimulating hormone [FSH] and/or human menopausal gonadotropin [hMG] after pituitary desensitization) and friendly IVF (no stimulation, clomiphene citrate, small dose of FSH or hMG without pituitary desensitization). The purpose of the present study was to evaluate the differences in steroid distribution between conventional and friendly IVF. Methods: Concentrations of estradiol, progesterone, FSH, and luteinizing hormone (LH) in serum and follicular fluid were determined in conventional and friendly IVF protocols by an enzyme-linked immunosorbent assay kit. Correlations between follicular fluid and serum steroid concentrations in these different protocols, and between pregnant cycles and steroid concentrations were evaluated. Results: Two hundred and thirty-four samples of follicular fluid from 74 IVF patients were analyzed. In conventional IVF, there was no relationship in steroid levels in between follicular fluid and serum steroids, whereas serum steroid concentrations correlated with the number of developing follicles. There was a relationship between the serum and follicular fluid estradiol levels (r = 0.467, P < 0.0001) as well as progesterone levels (r = 0.227, P = 0.0488) from friendly IVF patients. Conclusions: Serum steroid concentrations were mainly associated with the number of developing follicles. In the cases of friendly IVF, which had a small number of developing follicles, serum steroids might be used to monitor follicular fluid steroid concentrations. (Reprod Med Biol 2006; 5: 277-282).

Five cases of dizygotic triplet pregnancy following assisted reproductive techniques.

Of 816 patients who became clinically pregnant by assisted reproductive techniques between September 2000 and August 2004, we experienced 10 cases (1.2%) of monozygotic twinning, and in five of these 10 cases, implantation of another embryo resulted in dizygotic triplets. Here, we report these five cases of dizygotic triplets. Fresh embryo transfer was performed in all five cases. Intracytoplasmic sperm injection or assisted hatching was not carried out in these cases. Blastocyst transfer was performed in three cases. Three embryos were transferred in case 1 (40-year-old female). While only two embryos were transferred in the other four cases so as to avoid triplet pregnancy, triplet pregnancies were confirmed. Triplet pregnancy was maintained in three cases, but in the other two cases, monochorionic twinning resulted in miscarriage during the first trimester. For the three patients who delivered the triplets, while the postnatal growth has been normal for all nine babies, the mothers were hospitalized for a long period of time, and an emergency cesarean section was performed on two patients. Because triplet pregnancy could not be completely prevented even when only two embryos were transferred, physicians should be sure to obtain informed consent in similar cases. (Reprod Med Biol 2005; 4: 59-64).