Perinatal outcomes of patients who achieved pregnancy with a morphologically poor embryo via assisted reproductive technology.

PURPOSE: To learn more about the perinatal outcomes of conception via the use of low-grade embryos, we evaluated the relationship between the status of transferred embryos and the resultant perinatal outcomes. METHODS: A total of 340 patients who achieved pregnancy via ART treatment and consequently delivered in our clinic between April 2009 and March 2014 were recruited for this study. Patients were divided into two groups according to a morphological assessment of the transferred embryos, which relegated 79 patients into a poor-embryo group with the remainders (n = 261) placed into a good-embryo group. RESULTS: The mean maternal age for the poor-embryo group was 36.0 years, which was similar to the good-embryo group (36.8). In both groups, the percentages of fresh ET cycles were similar. The values for the mean birth weight and birth height of infants in the poor-embryo group were 3055 g and 50.3 cm, respectively, and there were no significant differences with the good-embryo group. The umbilical blood gas analysis in the poor-quality group was similar to that in the good-embryo group. There were no obvious major anomalies among the infants of either group. CONCLUSIONS: The perinatal outcomes of the poor-embryo group were comparable to those of the good-embryo group. Based on these results, we can provide qualified assurance for a normal perinatal outcome to patients who had no choice but to undergo embryo transfer with a poor-quality embryo.

Clomiphene citrate affects the receptivity of the uterine endometrium.

Purpose: To investigate whether clomiphene citrate (CC) affects uterine receptivity or not, we evaluated pregnancy rates (PR) during the hormone replacement cycle (HRC) according to the period between the last day of CC administration and the day of embryo transfer (ET). Methods: From March 2008 through March 2010, a total of 378 treatment cycles among 378 patients who received CC and had to avoid fresh ET due to a thin uterine endometrium were recruited. All patients underwent thawed ET using HRC. PRs were evaluated according to the period between the last CC treatment and the day of ET. Results: PR for the groups in which the period between the last CC treatment and the day of ET increased to more than 91 days were significantly higher than that for group in which the period was less than 90 days (p < 0.05). Conclusions: A lower PR was shown by the patients who underwent thawed ET in the HRC within 90 days after their last CC treatment, which shows that CC affects the receptivity of the uterine endometrium.

Prostaglandin therapy during the proliferative phase improves pregnancy rates following frozen embryo transfer in a hormone replacement cycle.

AIM: To determine the efficacy of prostaglandin administration during the proliferative phase in order to improve pregnancy rates following frozen embryo transfer during a hormone replacement cycle (HRC). METHODS: From September 2010 through March 2012, patients (n = 135) were recruited who had undergone oocyte retrieval during a stimulation cycle with clomiphene and had deferred fresh embryo transfer (ET) due to a thin uterine endometrium. All patients were less than 40 years of age and underwent thawed ET following all embryo cryopreservation, and were randomly divided into two groups for thawed ET using a conventional hormone replacement cycle with or without prostaglandin derivatives (prostaglandin or conventional group). Prostaglandin derivatives were administrated during the proliferative phase. Pregnancy and implantation rates following frozen ET were compared between the two groups. RESULTS: Although the endometrial thickness on the day of ET was similar for the prostaglandin and conventional groups, the pregnancy and implantation rates for the prostaglandin group were 40.0% and 22.0%, respectively, which was significantly higher than the rates for the conventional group (P < 0.01). CONCLUSION: Among patients who avoided fresh ET due to a thin endometrium, the pregnancy rate following a thawed cycle was low. However, it was improved when prostaglandin derivatives were used during the proliferative phase.

Advanced scheduling for zygote intrafallopian transfer is possible via the use of a hormone replacement cycle for patients who have experienced repeated implantation failures.

PURPOSE: Zygote intrafallopian transfer (ZIFT) is an effective option for patients who have experienced repeated implantation failures (RIF) in assisted reproductive technology (ART) treatment. However, advance planning for the day of the operation can be problematic. Using a hormone replacement cycle (HRC) makes it possible to plan for the day of ZIFT. In the present study, we evaluated whether HRC-ZIFT is useful for RIF patients who have experienced difficulties obtaining morphologically good embryos in vitro. METHODS: A total of 55 patients with a history of five or more unsuccessful transfers received HRC-ZIFT between June 2008 and June 2013. The oocyte pick-ups were performed and the oocytes showing two pronuclei (2PN) were cryopreserved. After receiving more than five 2PN oocytes, the operation day was scheduled in advance, and as a consequence, a HRC was started and ZIFT was performed. The clinical outcomes were evaluated. RESULTS: The average age of the patients was 39.3 years, and the previous OPU and ET attempts numbered 7.5 and 6.9, respectively. The number of previously transferred embryos was 11.8, and the number of morphologically good embryos (MGEs) was only 1.2. The number of transferred 2PN oocytes was 6.7, and the subsequent pregnancy rate was 23.6 %. No ectopic or multiple pregnancies were observed, but there were 6 cases of miscarriage. CONCLUSION: Among RIF patients, in particular those who have difficulty obtaining MGEs in vitro, ZIFT might be a useful option. The HRC allows patients and medical staff to plan for the operation day in advance.

Role of salpingoscopy in assessing the inner fallopian tubes of infertility patients with ovarian endometriomas.

AIM: The aim of this study was to observe the insides of the fallopian tubes of patients with unilateral or bilateral endometriomas by using salpingoscopy and evaluate the inner cavity of the fallopian tubes according to our original scoring system. MATERIAL AND METHODS: From April 2008 through December 2010, patients with unilateral or bilateral endometriomas were recruited (n = 157, endometrioma group). All patients underwent laparoscopic ovarian cystectomy and salpingoscopy. Using salpingoscopy, we observed the tubal lumen and calculated a fallopian tube score (F score) paying attention to the following six results: adhesions, loss of mucosal folds, rounded edges of mucosal folds, debris, foreign bodies, and abnormal vessels. The F scores were compared with those of the unexplained infertility patients who received those same procedures during the same period (n = 235; control group). RESULTS: Slightly more than three-quarters (75.9%) of the patients in the endometrioma group received F scores of 0, and this percentage was significantly higher than that for the control group (139/235 = 59.1%, P < 0.05). The pregnancy rate after conventional treatment for the endometrioma group was 21.7%, and all pregnant patients had achieved an F score of less than 2. CONCLUSION: It is highly possible that infertility patients with ovarian endometriomas are more likely to have intact fallopian tubes, by comparison with infertility patients who do not have ovarian endometriomas.

Using a mild stimulation protocol combined with clomiphene citrate and recombinant follicle-stimulating hormone to determine the optimal number of oocytes needed to achieve pregnancy and reduce the concerns of patients.

PURPOSE: The purpose of this study was to investigate how many oocytes are needed to achieve an adequate pregnancy rate per 1 oocyte retrieval cycle in mild ovarian stimulation. METHODS: This protocol consisted of clomiphene citrate and recombinant-follicle-stimulating hormone injection without a gonadotropin-releasing hormone-antagonist. From January 2009 through December 2010, there were 1,227 women who underwent assisted reproductive technologies treatment with mild stimulation at the Sugiyama Clinic. The overall pregnancy rate per single oocyte retrieval cycle was evaluated using both fresh and cryopreserved-and-thawed embryos according to the retrieved oocyte number. RESULTS: According to the retrieved oocyte number, a total of 1,227 cycles were divided into 4 groups: group A (the oocyte number <4; 433 cycles), group B (the oocyte number = 4, 5; 317 cycles), group C (the oocyte number = 6, 7; 206 cycles), and group D (the oocyte number ≥8; 271 cycles). The overall pregnancy rates for groups A, B, C, and D were 22.2, 42.9, 52.4, and 56.0 %, respectively, the rates for groups C and D were significantly higher than that for group A (p < 0.01). CONCLUSIONS: The optimal number of retrieved oocytes proved to be between 6 and 7 for the patients who received our milder stimulation protocol and experienced no reduction in their overall pregnancy rate.

Hyaluronan-enriched transfer medium improves outcome in patients with multiple embryo transfer failures.

PURPOSE: To ascertain whether the use of hyaluronan-enriched transfer medium (HETM) improves pregnancy and implantation rates among embryo transfer patients with a history of multiple implantation failures. METHODS: Patients (n = 314) under the age of 40 and with a history of multiple unsuccessful embryo transfers were enrolled. There were three groups of patients: those undergoing fresh embryo transfer (fresh ET [n = 111]), those undergoing vitrified-warmed ET in the natural cycle (WET-N [n = 101]) and those undergoing WET in a hormone replacement cycle (WET-H [n = 102]). On the day of ET, patients were randomized to HETM (0.5 mg/ml hyaluronan) or control medium containing no hyaluronan. Only patients with good quality embryos on day 3 were included. RESULTS: For all three patients groups (fresh ET, WET-N and WET-H) pregnancy rates (37.5 %, 31.4 % and 41.2 %, respectively) were significantly higher when using HETM compared with control medium (10.9 %, 10.0 % and 15.7 %, respectively; p < 0.05), and implantation rates when using HETM were also significantly higher compared with control medium (p < 0.05). Miscarriage rates were similar in both groups. CONCLUSION: HETM significantly increased pregnancy and implantation rates among embryo transfer patients with a history of multiple unsuccessful implantations-regardless of method used to prepare the endometrium.

A programmed schedule of oocyte retrieval using mild ovarian stimulation (clomiphene citrate and recombinant follicle-stimulating hormone).

PURPOSE: The purpose was to establish a mild ovarian stimulation protocol that would help assisted reproductive technology (ART) units to avoid scheduling on weekends. METHODS: This protocol directed patients to take 50 mg/day of clomiphene citrate between days 3 and 7 of the menstrual cycle: 225 IU of recombinant follicle-stimulating hormone (rec-FSH) were administered on days 3, 5 and 7; human chorionic gonadotropin (hCG) was administered on day 9; and, oocyte pick-up (OPU) was planned for day 11. From October 2008 through October 2009, 514 women underwent ART treatment with mild stimulation at the Sugiyama Clinic, and we evaluated whether OPU was accomplished on the planned day. RESULTS: Of all the treatment cycles, 419 (81.5%) underwent OPU on day 11 (scheduled group). Additional rec-FSH administration was needed in 83 cycles, in which case OPU was performed on day 12 or later. In 12 cycles, OPU was canceled. The unscheduled group (n = 95) consisted of delayed OPU cycles and canceled cycles. Of all treatment cycles, 332 cycles in the scheduled group and 68 cycles in the unscheduled group underwent embryo transfer, with 81 and 16, respectively, resulting in pregnancies. CONCLUSIONS: Using this protocol, OPU was performed on the scheduled day in about 80% of the cycles. Most weekend scheduling of OPU can be avoided using this mild stimulation.

Elective single cleavage-stage embryo transfer need not result in lower pregnancy rates compared to double cleavage-stage embryo transfer.

AIM: To confirm whether women who choose to have one fresh embryo transferred and one frozen-and-thawed embryo when needed can dramatically reduce the possibility of a multifetal pregnancy while giving themselves a better chance of achieving pregnancy. METHODS: We enrolled 685 patients who were undergoing assisted reproductive technology (ART) treatment at our clinic between January 2005 and December 2008. None of the patients had a history of ART treatment, and they received either a double-embryo transfer (DET) or single-embryo transfer (SET) during this period. The outcomes of the ART and the pregnancy rates per patient were evaluated for both groups and comparisons were made. RESULTS: The mean age was 35.7 +/- 0.2 years (mean +/- standard error of the mean) for all patients (n = 583) who received a fresh embryo cycle of DET. In contrast, the mean age (34.3 +/- 0.4) of all patients (n = 102) who received a fresh- or thawed-embryo transfer cycle of SET was significantly younger than the average age in the DET group (P < 0.05). The per-patient overall pregnancy rate in the SET group was an estimated 35.3%, which was significantly higher than that in the DET group (P = 0.02). However, the multifetal pregnancy rate for the DET group was significantly higher than that for the SET group (P < 0.01). CONCLUSION: We demonstrated that women who choose to have one fresh embryo transferred and one frozen-and-thawed embryo when needed, can dramatically reduce their possibility of a multifetal pregnancy while giving themselves a better chance of achieving pregnancy.

Clinical outcomes resulting from the transfer of vitrified human embryos using a new device for cryopreservation (plastic blade).

PURPOSE: We attempted clinical application of a plastic blade, which is a novel cryopreservation device, for vitrification of human embryos and blastocysts. METHODS: Between February 2003 and December 2007, a total of 4,430 Day 3 embryos from 898 patients (Day 3 group) and 55 blastocysts from 29 patients (blastocyst group) were vitrified and cryopreserved with a plastic device, and subsequently thawed for embryo transfer. Clinical outcomes after thawing and transfer of vitrified embryos and blastocysts were evaluated. RESULTS: In the Day 3 group, all embryos resulting from 1,441 oocyte retrieval cycles were recovered, and the thawed embryo survival rate was 98.4%. In the blastocyst group, the survival rate after thawing was 100%. A total of 3,026 day 3 embryos and 46 blastocysts were transferred. The pregnancy and implantation rates in the Day 3 group were 25.0% and 15.5%, respectively, and in the blastocyst group the rates were 24.2% and 26.1%, respectively. The miscarriage rates in the Day 3 and blastocyst groups were 18.3% and 50.0%, respectively. CONCLUSIONS: A plastic blade is a useful novel device in cryopreservation of vitrified human embryos.

A study of the effect of an extremely low oxygen concentration on the development of human embryos in assisted reproductive technology.

Purpose: To determine whether embryos cultured with a low oxygen level (2%) brought about beneficial effects on the outcome of ART. Methods: This is a sequential case-control embryo-culture study. Embryos were cultured either with a gas mixture containing 2% O2, 5% CO2, and 93% N2 (low-oxygen group) or 5% O2, 5% CO2, and 90% N2 (conventional group). From January 2008 to September 2008, 873 fertilized oocytes were obtained from 250 patients in the low-oxygen group and from October 2008 to March 2009, 730 fertilized oocytes were obtained from 213 patients in the conventional group. The outcomes of ART were compared between two groups. Results: The cleavage rate in the low-oxygen group (94.4%) was similar to that (94.7%) in the conventional group. The mean number of blastomeres on Day 3 in the low-oxygen group (mean ± SE) was 6.5 ± 1.9, and this was significantly lower than in the conventional group (6.8 ± 1.9, p < 0.05). Moreover, the low-oxygen group produced worse quality embryos, on the basis of the significantly higher embryo grade 2.1 ± 0.6 versus 1.9 ± 0.6, p < 0.001, in 5% oxygen. The pregnancy and miscarriage rates in the low-oxygen group were 22.3 and 20.8%, respectively, which were statistically similar to the outcomes in the conventional group. Conclusions: Overall, culture of embryos at the low oxygen level did not significantly improve ART results compared with embryos grown in 5% oxygen. The study suggests that a low oxygen level worsens embryo morphology but does not impair embryo viability.

The dilemma faced by patients who undergo single embryo transfer.

PURPOSE: The aim of this study was to identify the factors that contribute to the decision to choose single embryo transfer (SET). METHODS: Two hundred and nine patients who underwent ART treatment in our clinics between April 2006 and May 2007 were enrolled in this study. All patients had elected to undergo SET before the start of each treatment cycle; a questionnaire was administered to all patients prior to the SET procedure. RESULTS: The mean age of the patients was 34.6 years old (range: 24-45 years). The mean number of redundant embryos was 3.7 (range: 1-17), and the pregnancy rate per embryo transfer was 25.7%. A total of 121 patients (57.9%) who underwent SET returned their questionnaires. Based on the results of questionnaire, 56.2% of patients who received SET waived their right to choose between single and double embryo transfer. Among patients who selected SET, 67.6% believed that the pregnancy rate resulting from double embryo transfer (DET) is significantly greater than that associated with SET, and 25% of patients wanted to have twins. The majority of patients (80.9%) who underwent SET understood that multi-fetal pregnancy increases the risk of complications during gestation and delivery. Among all patients who completed the questionnaire, 72.8% believed that the number of transferred embryos should not be controlled by law. CONCLUSIONS: The results of the present study show that greater than one-half of patients who underwent SET were faced with a dilemma--the difficult choice between their own desires and their clinician's recommendation.

Gasless laparoscopic surgery using a new intra-abdominal fan retractor system: an experience of 500 cases.

AIM: The aim of this study is to report the feasibility of a newly developed intra-abdominal fan retractor system for use in gynecologic laparoscopic surgery. METHODS: Five hundred women undergoing gasless laparoscopic surgery using the abdominal wall lifting device were included in the study. The intraoperative and postoperative courses, and complications were examined. RESULTS: The intra-abdominal retractor system provided adequate exposure in all cases, except for one patient with morbid obesity. Neither the presence of the intra-abdominal retractor blades nor the mechanical arm interfered with the placement of instruments during surgery. No complications related to the use of gasless laparoscopy were encountered in this study period. CONCLUSION: The new intra-abdominal fan retractor system is feasible in gynecologic laparoscopic surgery.

The Effect of a Newly Designed Needle on the Pain and Bleeding of Patients During Oocyte Retrieval of a Single Follicle.

BACKGROUND: The purpose of this study was to evaluate the effect of needle type on pain and bleeding during oocyte pick-up (OPU). METHODS: From May through November 2013, patients undergoing OPU from a single follicle without any analgesic treatment were including this study. Eligible patients (n=75) were randomized 1:1 to undergo the procedure with either a reduced needle (17 gauge body, 20 gauge tip; RN group) or a standard needle (19 gauge; SN group). Overall pain was assessed by patients using a visual analogue scale (VAS), and vaginal bleeding after the procedure was recorded. Fisher exact, t-test or Wilcoxon test were used, and p<0.05 was considered to be statistically significant. RESULTS: The percentage of mature oocytes was 86.5% in the RN group and 91.7% in the SN group. Pain during OPU was significantly lower in the RN group than in the SN group (mean VAS score±SD: 3.2±2.0 cm vs. 4.9±2.2 cm, p<0.01; mean±SD). The frequency of vaginal bleeding was also significantly lower in the SN group (26.3% vs. 48.6%; p<0.05). The frequency of bleeding in the RN group was also significantly lower than that in the SN group (26.3% vs. 48.6%; p<0.05). No significant differences were found between the two groups with regard to fertilization and pregnancy rates. CONCLUSION: The newly designed needle significantly reduced pain and vaginal bleeding associated with single-follicle OPU in patients receiving no analgesic treatment, in comparison with a standard needle. The RN had no adverse effect on the quality of retrieved oocytes.

Change of the mitochondrial distribution in mouse ooplasm during in vitro maturation.

Mitochondria (mt) have been reported to be closely related to the maturation of mammalian oocytes, but their function in oocyte maturation has not been elucidated. In this study, we examined the kinetics of mt and chromatin configuration during in vitro maturation of mouse oocytes to clarify the relationship between oocyte maturation and mitochondrial distribution morphologically. Oocytes were recovered from 6-to 8-wk-old ICR strain female mice. Germinal vesicle (GV) -stage oocytes were divided into 4 groups and cultured: group A, oocytes collected after pregnant mare serum gonadotropin (PMSG) injection; and group B, oocytes collected after PMSG-human chorionic gonadotropin injection. Groups A and B were subdivided into 2 groups: denuded oocytes (DO) and cumulus-enclosed-oocytes (CEO). At 0, 4, 8, 12 and 16 h from the onset of the culture, oocytes were fixed and stained to visualize alpha-tubulin, chromatin and mt using confocal laser scanning microscopy (CLSM). It was observed that mt aggregated around the nucleus from the GV-stage through progression to germinal vesicle breakdown (GVBD). With the movement of the nucleus, mt were concentrated around the nucleus and polarized. The maturation rate (the rate of the first polar body extrusion) and the fertilization rate of CEO were significantly higher than that of DO in both groups A (p<0.01) and B (p<0.05). During the GV-stage to GVBD, the rate of mitochondrial aggregation around the nucleus tended to be high in group A (CEO). The rates of mitochondrial polarization in MI and MII oocytes were 76.1% with in-vitro maturation (IVM) and 86.7% with in-vivo-maturation, respectively; the rate was significantly higher in in-vivo-maturation-oocytes than in IVM-oocytes (p<0.01). From the present results it can be considered that aggregaton of mitochondria around the nucleus was essential for maturation, fertilization and development.

A new evaluation score that uses salpingoscopy to reflect fallopian tube function in infertile women.

OBJECTIVE: To build an evaluation scoring system using the results of salpingoscopy, and to evaluate the relationship between this scoring system and the outcome of pregnancy. DESIGN: Retrospective study. SETTING: Sugiyama Clinic. INTERVENTION(S): Using salpingoscopy, we observed the tubal lumen, paying attention to the following six results: adhesions, loss of mucosal folds, rounded edges of mucosal folds, debris, foreign bodies, and abnormal vessels. PATIENT(S): From April 2008 through June 2009, 104 women in whom unexplained infertility had been diagnosed underwent salpingoscopy. The F scores were evaluated related with various clinical results or pregnancy rates. MAIN OUTCOME MEASURE(S): The F score expressed the sum of the abnormal results, and one abnormal result was given a 1-point F score. RESULTS: Approximately 60% of the patients showed an F score of 0, and the percentages of patients who showed 1, 2, 3, and ≥4 points were 19.2%, 11.5%, 4.5%, and 4.5%, respectively. After evaluation, 23 patients achieved pregnancy within a year. The pregnancy rates for patients with F scores of 0 and 1 point were 30.6% and 20.0%, respectively, and the rate of patients with an F score of 0 was significantly higher than the rate of patients with high F scores (F score ≥2; 9.1%). CONCLUSION(S): The patients showing a lower F score (0 or 1) showed higher fecundity than those showing an F score of ≥2.

Effect of sera on the adhesion of natural killer cells to the endothelium in severe pre-eclampsia.

OBJECTIVE: To investigate the effect of serum on the interaction between natural killer (NK) cells and endothelial cells in pre-eclampsia. METHODS: Seven severely pre-eclamptic patients, five normal pregnant women, and four normal non-pregnant women were included in this study. Freshly isolated NK cells labeled with Chromium-51 were incubated on an endothelial cell monolayer in the presence of patient serum. In regard to the characteristics of adhesive molecules, the endothelial cells were blocked by monoclonal antibodies (mAbs) to intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1); the NK cells were blocked by mAbs to leukocyte function-associated antigen (LFA-1) and very late antigen-4 (VLA-4) before co-incubation. After incubation, the adherent cells were solubilized with 1% Triton X. The lysates were collected and counted in a gamma counter. RESULTS: The adhesion of NK cells to the endothelium in the normal pregnancy group decreased significantly in comparison to the non-pregnant group (7%vs 72%; P < 0.01). Adhesion in the severe pre-eclamptic group was significantly higher in comparison to the normal pregnant group (44%vs 7%; P < 0.01). The blocking percentages of mAbs on NK adhesion in the severe pre-eclampsia group were 49 +/- 4% to LFA-1, 61 +/- 48%, 67 +/- 39% to VLA-4, ICAM-1, and 68 +/- 7% to VCAM-1. CONCLUSION: Sera from normal pregnant women suppress the adhesion between NK cells and endothelial cells, whereas the suppressive effect of sera from pre-eclamptic patients has a diminished affect.