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Case report: pregnancy outcome following ICSI of oocytes with abnormal cytoplasm and zona pellucida.

A couple with unexplained infertility was referred for routine IVF and ICSI treatment. Ovulation was induced by the GnRH analogue protocol combined with HMG and HCG. Preparation of denuded oocytes revealed a major disorder of the zona pellucida and abnormal oocytes. During preparation of ova for ICSI, 15 retrieved oocytes were denuded, 14 of which underwent ICSI treatment. Four of the oocytes collapsed and the remaining 10 appeared to have irregular, fragile zona pellucida. Nevertheless, following ICSI, seven low-quality embryos developed, three of which were transferred into the uterus. Two implantations were achieved, but only one embryo resulted in an uneventful pregnancy and delivery by Caesarean section of a normal female neonate with an Apgar score of 10. It is hypothesized that infertility was due to the abnormal oocyte structure and abnormal zona pellucida, which prevented natural conception. This condition was successfully resolved by the ICSI procedure.

Adult↗

[Use of GnRH antagonists (cetrorelix) in assisted reproduction: first report in Mexican literature].

The use of GnRH antagonists in assisted reproduction techniques (ARTs) has been shown to be effective in blocking the luteinizing hormone (LH), without a repercussion on ovaric response. The present study reports the use of a GnRH antagonist (cetrorelix) 0.25 mg/d s.c. in patients stimulated with recombinant FSH (rFSH) for IVF/ICSI. 44 patients with mean age of 33.3 years were included in this study, four patients were receptors for oocyte donation program and another patients was receptor for frozen embryos. Controlled ovarian stimulation was carried out with rFSH, starting on day 3 of menstrual cycle. The total mean dose used was 3.570 +/- 1.102 UI for each patient. The LH serum concentrations were monitored during ovaric stimulation, in two patients a premature LH surge (> 10 UI/L) was detected. The oocytes were collected 36 hours after hCG administration, the mean preovulatory oestradiol was 4.006 +/- 2.418 pg/mL. The mean total oocytes retrieved was 10.8 +/- 7.4 per patient, the fertilization rate was 58.3%, in only three patients fertilization was failed. No difference was observed in oocyte retrieval, fertilization rate, implantation rate between both groups (GnRH antagonists versus GnRH agonists). The embryo transfer was realized on day 3, and only two patients was on blastocyst stage. In 17 of 41 patients with successful transfer, pregnancy was achieved in the group with antagonists, and 8 of 20 patients in the group with agonists. In conclusion Cetrorelix prevents adequately premature LH surges and allows an effective ovaric response in controlled stimulation in IVF cycles, except in patients in oocyte donation programs who require higher cetrorelix doses.

Adult↗

Elective single day 3 embryo transfer halves the twinning rate without decrease in the ongoing pregnancy rate of an IVF/ICSI programme.

BACKGROUND: Data on the effect of elective single embryo transfer (eSET) on the total and multiple pregnancy rates of an IVF/ICSI programme are reported. METHODS AND RESULTS: A retrospective cohort analysis of eSET was carried out over a 4 year period. A total of 1559 cycles resulted in 1464 transfers; 299 transfers of one top quality embryo (20.4%) and 86 of one non-top quality embryo (5.9%) yielded 149 conceptions (49.8%) with 105 ongoing pregnancies (35.1%) and 26 conceptions (30.2%) with 19 ongoing implantations (22.1%) respectively; 1079 transfers of two (n = 853; 58.3%) or more than two (n = 226; 15.4%) embryos yielded 366 ongoing pregnancies (33.9%). The ongoing pregnancy rates for the years between 1998 and 2001 were 35.9, 27.9, 31.9 and 31.0% per oocyte retrieval and 38.5, 29.4, 34.1 and 33.2% per transfer. There were no differences in pregnancy rates between any of the years. The average ongoing pregnancy rate (>12 weeks) over the 4 years was 31.5% per started cycle and 33.5% per transfer; the average number of embryos transferred decreased from 2.26 (1998) to 1.79 (2001); the multiple pregnancy and twinning rates dropped from 33.6 and 29.5% (1998) to 18.6 and 16.3% (2001) respectively. CONCLUSIONS: Judicious application of eSET can halve the twinning rate while maintaining the overall pregnancy rate.

Adult↗

Serum PAPP-A levels at 10-14 weeks of gestation are altered in women after assisted conception.

OBJECTIVES: Increased screen-positive rate was observed in the first-trimester screening for trisomy 21 (T21) among women after assisted conception. The aim of this study was to identify the trends of changes in serum marker levels after different modes of assisted conception. METHODS: Retrospectively, we analyzed the levels of free beta human chorionic gonadotrophin (f beta HCG), pregnancy-associated plasma protein-A (PAPP-A) and inhibin A in maternal serum, and fetal nuchal translucency (NT) thickness at 10-14 gestational weeks in 1098 women with singleton pregnancies: 130 after IVF-ET, 54 after ICSI and 914 after spontaneous conception. RESULTS: In women after IVF-ET and ICSI, PAPP-A was decreased (0.94 MoM and 0.82 MoM, respectively) and inhibin A increased (1.11 MoM and 1.48 MoM, respectively) in comparison to the women after spontaneous conception. With increasing number of oocytes retrieved PAPP-A decreased and inhibin A increased. In women pregnant after assisted conception without ovarian stimulation (transfer of frozen-thawed embryo or in spontaneous cycle) the marker levels were not significantly different from those in women after spontaneous conception. CONCLUSIONS: The presence of multiple corpora lutea may be responsible for the changes in marker levels. In spite of the differences in marker levels, the screen-positive rate for T21 is not significantly increased in women after assisted conception.

Adult↗

Testicular biopty gun needle biopsy in collecting spermatozoa for intracytoplasmic injection, cryopreservation and histology.

Using testicular spermatozoa from either open biopsy (29 cycles) or biopty gun needle biopsy (49 cycles), a total of 81 intracytoplasmic sperm injection (ICSI) cycles among 57 couples were carried out from January, 1994 to September, 1997. In six cycles, no spermatozoa were obtained, and in three cycles spermatozoa from both needle and open biopsies were used. The fertilization (37% after open and 41% after needle biopsy) and pregnancy rates (29% per embryo transfer compared with 16% per embryo transfer) were similar after both open and needle biopsies. Five pregnancies were achieved among the 14 couples with non-obstructive azoospermia of the male partner, four of these after needle biopsy. It was possible to use cryopreserved testicular spermatozoa after both needle and open biopsies, and one pregnancy started after using cryopreserved testicular spermatozoa in both groups. Histological needle biopsy was carried out in 62 cases, and they were all diagnostic, giving 15-20 cross-sections of seminiferous tubuli per biopsy. Testicular needle biopsy using a 14 gauge biopsy needle gave a sufficient amount of tissue and spermatozoa for ICSI, cryopreservation and histology, even in non-obstructive azoospermia. This technique is simpler and cheaper than open biopsy and, hence, it can be regarded as the optimal method for the retrieval of testicular spermatozoa.

Adult↗

Evaluation of 225 patients undergoing subzonal insemination for the procurement of fertilization in vitro.

OBJECTIVE: To evaluate subzonal insemination for the procurement of fertilization in infertile males. DESIGN: The spermatozoa of infertile males was used, where possible, for in vitro fertilization (IVF) and subzonal insemination with the same cohort of oocytes obtained from their partner. SETTING: An individual unit in a general hospital, Clinical Nomentana, Rome, Italy. PATIENTS: Males who had repeatedly failed to achieve conception in vitro by IVF or in whom seminal parameters were unacceptable for IVF were treated. INTERVENTIONS: Two hundred twenty-five transvaginal oocyte retrievals were performed. MAIN OUTCOME MEASURES: The incidence of fertilization, cleavage, and pregnancy was evaluated in relation to the numbers of spermatozoa injected and the individual spermatozoa parameters. RESULTS: Fertilization occurred in 39% of patients and 16% of eggs, and 12 clinical pregnancies were established. Fertilization increased with increasing numbers of spermatozoa injected. Fertilization with subzonal insemination was significantly greater (39%) than after in vitro insemination (6%) (P less than 0.0001). CONCLUSIONS: Subzonal insemination improved the incidence of fertilization in this series of patients, and a new classification of spermatozoa based on total motile count aided prognosis for fertilization.

Adult↗

Intracytoplasmic sperm injection: correlation of oocyte grade based on polar body, perivitelline space and cytoplasmic inclusions with fertilization rate and embryo quality.

The extent to which the morphology of the oocyte at the light microscopy level is related to the results of intracytoplasmic sperm injection (ICSI) is controversial. In this study, after cumulus removal, oocytes were graded into four groups according to the status of the first polar body, size of the perivitelline space and the presence of cytoplasmic inclusions. Oocyte data from 65 consecutive patients were reviewed. The results showed that, for oocytes without cytoplasmic inclusions, the fertilization rate and embryo development beyond 2-cell stage were significantly lower (P < 0.01) in the oocytes at grade 1-2 (poor) than those in oocytes at grade 3-4 (good). Grade 4 oocytes without inclusions gave the highest proportion (66.7%) of good embryos with grading 1-2 (grade 1 best; P < 0.01). A higher proportion of grade 1-2 oocytes (44.7%; P < 0.05) was obtained from patients older than 35 years. More oocytes containing cytoplasmic inclusions were seen in patients diagnosed as having female factor infertility (24.9%; P < 0.01) and older than 35 years (26.5%; P < 0.05) compared to patients with male factor infertility and younger than 35 years. The fertilization rate and embryo development were not associated with the oestradiol concentration on the day of human chorionic gonadotrophin administration or the total number of oocytes retrieved. The results suggest that human oocyte grading based on the triple factors first polar body, size of perivitelline space and cytoplasmic inclusions is related significantly to fertilization rate and embryo quality after ICSI.

Adult↗

Reproductive performance after ultrasound-guided transvaginal ethanol sclerotherapy for ovarian endometriotic cysts.

OBJECTIVE: Ultrasound-guided transvaginal ethanol sclerotherapy (TV-EST) has been widely practised in Japan for ovarian endometriotic cysts. We investigated the possible adverse effects of TV-EST for ovarian endometriotic cysts on reproductive performance. PATIENTS AND METHODS: We reviewed retrospectively medical records and compared clinical outcomes of 45 subfertile women who underwent TV-EST for ovarian endometriotic cysts (Study group) with those of 65 subfertile women without ovarian endometriotic cysts (Comparison group). Patients were followed up monthly for 24 months. Serum levels of LH, FSH, CA125 and CA19-9 were determined before and after TV-EST. RESULTS: No complications associated with TV-EST were observed. There were no differences in the numbers of pregnancies (47% (21/45) versus 39% (25/65)), term deliveries (76% (16/21) versus 76% (19/25)), abortions (19% (4/21) versus 24% (6/25)), retrieved oocytes, or quality of embryos between the Study and Comparison groups, respectively. The serum levels of LH and FSH did not increase after TV-EST. The serum levels of CA125 and CA19-9 did not significantly decrease after TV-EST. Ovarian cysts recurred in six (13.3%) of the 45 women 5.2+/-3.9 months after TV-EST. CONCLUSION: Although only a small number of women were studied, our observational study suggested that TV-EST appeared not to adversely affect reproductive performance in subfertile women with ovarian endometriotic cysts.

Adult↗

Coarse granulation in the perivitelline space and IVF-ICSI outcome.

PURPOSE: To study the effect of the presence of coarse granules in the perivitelline space (PVS) of oocytes on embryonic development, and on implantation and pregnancy rates in IVF. METHODS: The study population included 24 patients treated during the period 1995-2000. The majority or all of their oocytes exhibited repeatedly coarse granulation in the PVS. Clinical and laboratory cycle characteristics of their 65 IVF-ICSI cycles and the resulting implantation and pregnancy rates were compared to a matched control group of 65 IVF- ICSI cycles without granulation in the PVS. RESULTS: A total of 623 oocytes were retrieved, 418 oocytes fertilized, and 246 embryos were transferred in the study group. No difference was detected between the study and control group with regard to patients' clinical data, IVF cycle characteristics, mean number of oocytes retrieved and fertilized, and mean number of embryos transferred. Only seven pregnancies were achieved in the study group, leading to pregnancy and implantation rates of 10.7 and 5.7%, respectively. Pregnancy and implantation rates were significantly higher in the control group of matched IVF-ICSI cycles without granulation in the PVS (32.5 and 11.5%, respectively). CONCLUSIONS: The presence of coarse granules in the PVS correlates with low implantation and pregnancy rates in IVF-ICSI cycles and might be regarded as a distinct entity, part of the yet poorly defined condition of "egg factor infertility."

Adult↗

Oocyte morphology predicts outcome of intracytoplasmic sperm injection.

To examine the influence of cytoplasmic morphology on the success rate of intracytoplasmic sperm injection (ICSI), the morphology of 837 metaphase II oocytes was assessed after cumulus stripping. The main abnormalities detected were excessive granularity, cytoplasmic inclusions such as vacuoles, smooth endoplasmic reticulum clustering and refractile bodies. Microinjection was performed in 538 oocytes with normal cytoplasm, 142 out of 161 with excessive granularity and 112 out of 138 with cytoplasmic inclusions. Very poor oocytes were not injected. No difference was found in fertilization rate. The embryos achieved cleaved normally and a similar number of good quality embryos among the three groups was noted. The outcome of transfer of embryos derived solely from normal oocytes (group A: 72 patients, 183 embryos) was compared with those from oocytes with cytoplasmic abnormalities (group B: 34 patients, 85 embryos). In group A, 17 clinical pregnancies (24% per patient, implantation rate 10%) were established. In group B, only one clinical pregnancy (3% per patient, implantation rate 1%) was established, from the transfer of embryos derived from oocytes with homogeneous granularity of the cytoplasm. No pregnancy resulted following the transfer of embryos from eggs with cytoplasmic inclusions. The difference was statistically significant. The outcome of ICSI is dependent on the quality of the oocytes retrieved. Normal fertilization and early embryo development were achieved in oocytes with abnormal cytoplasm morphology, but the resulting embryos failed to demonstrate the same implantation potential as those derived from oocytes with normal cytoplasm.

Adult↗

Assessment of the ovarian volume, number and volume of follicles and ovarian vascularity by three-dimensional ultrasonography and power Doppler angiography on the HCG day to predict the outcome in IVF/ICSI cycles.

OBJECTIVE: The aim of this prospective study was to investigate whether ovarian blood flow is related to embryological parameters and whether it could be a predictor of outcomes of IVF/ICSI. METHODS: Eighty infertile women underwent ovarian stimulation with gonadotrophins after a long protocol with GnRH agonists. The ovarian volume (OV), number of follicles (NF) and follicular volume (FV) of all follicles >10 mm and vascularization index (VI), flow index (FI) and vascularization-flow index (VFI) were obtained by three-dimensional (3D) ultrasonography and power Doppler angiography (PDA) on the day of HCG administration. These parameters were tested for their relation with IVF laboratory parameters. RESULTS: The OV, FV, VI, FI and VFI were significantly greater in the pregnant group. The NF and FV were the only independent predictors of the number of oocytes retrieved, mature and fertilized, and the number of embryos developed and their cumulative embryo score. Nevertheless, the number of grade 1 embryos depends on the NF and the VI. The ovarian FI and the number of transferred grade 1 embryos can predict gestation in 76% of IVF patients. A low FI and non-grade 1 embryo transferred are also associated with an increased pregnancy loss. CONCLUSION: 3D ultrasonography and PDA allow for an easier ovarian assessment in IVF cycles. The predictive value of IVF outcome suggests a high clinical usefulness of this new technique.

Adult↗

Shortened exposure of oocytes to spermatozoa improves in-vitro fertilization outcome: a prospective, randomized, controlled study.

A prospective, randomized study of 158 patients undergoing in-vitro fertilization (IVF) and embryo transfer was conducted to evaluate whether a shortened exposure of oocytes to spermatozoa enhances oocyte development, and subsequently influences the IVF outcome. A comparison was made between conventional treatment time and shorter exposure of retrieved oocytes to spermatozoa. Fertilization and cleavage rates, embryo quality, implantation and pregnancy rates in the study group (short exposure) versus controls (standard IVF procedure) were evaluated. Fertilization (56 versus 61%) and cleavage rates (96 versus 92%) were similar in the two groups respectively. However, embryo quality was significantly higher in the study group (P < 0.05). Moreover, the pregnancy and implantation rates were significantly increased (42.4 versus 26% per embryo transfer, and 16 versus 10% respectively; P < 0.05). Our results demonstrated that shorter exposure of oocytes to spermatozoa is superior to the standard time in IVF and may have a favourable effect on implantation rates by improving embryo quality.

Adult↗

Comparison of pregnancy outcomes of progesterone or progesterone + estradiol for luteal phase support in ICSI-ET cycles.

OBJECTIVE: To find out the effect of estradiol with progesterone for luteal phase support in IVF-ICSI cycles. MATERIALS AND METHODS: Patients were accepted for treatment in the ART unit of Selcuk University, Meram Faculty of Medicine, between January 2001 and March 2003. The study was done in a prospective manner. The age range of 252 women was 19-41 years and the total number of cycles was 310. All patients were treated with a long ovulation induction protocol. Patients were treated and divided into two groups in a randomized manner: group I used only 600 mg/day divided into three equal doses of micronized progesterone vaginally, and group II used transdermal estradiol 100 microg/day + 600 mg/day vaginal micronized progesterone. RESULTS: 310 ICSI cycles were carried out in 252 infertile couples between January 2001 and March 2003. From 22 of these cycles, oocytes were retrieved but no embryos were developed. In the remaining 288 cycles there were embryo transfers. All embryo development was achieved by ICSI treatment. In 148 out of 288 cycles, the luteal phase was supported only by vaginal micronized progesterone (group I). On the other hand, the remaining 140 cycles received vaginal micronized progesterone plus transdermal estradiol 100 microg/day (group II). The number of beta-hCG-positive results in group I and group II were 20 (13.5 %) and 54 (38.5%) respectively. CONCLUSION: Adding estradiol to progesterone for luteal phase support in ICSI-ET cycles may increase implantation and pregnancy rates.

Adult↗

Maturation arrest of human oocytes as a cause of infertility: case report.

Maturation arrest of human oocytes may occur at various stages of the cell cycle. A total failure of human oocytes to complete meiosis is rarely observed during assisted conception cycles. We describe here a case series of infertile couples for whom all oocytes repeatedly failed to mature during IVF/ICSI. Eight couples, all presenting with unexplained infertility, underwent controlled ovarian stimulation followed by oocyte retrieval and IVF/ICSI. The oocytes were stripped of cumulus cells prior to the ICSI procedure and their maturity status was defined. In each couple, oocyte maturation was repeatedly arrested at the germinal vesicle (GV) (n = 1), metaphase I (MI) (n = 4) and metaphase II (MII) (n = 3) stage. Oocyte maturation arrest may be the cause of infertility in some couples previously classified as having unexplained infertility. The recognition of oocyte maturation arrest as a specific medical condition may contribute to the characterization of the yet poorly defined entity currently known as 'oocyte factor'. The cellular and genetic mechanisms causing oocyte maturation arrest should be the subject of further investigation.

Adult↗

In vitro fertilization. The Jamaican experience.

In June 2000, twenty-eight infertile couples were treated by in vitro fertilization and embryo transfer at our initial assisted reproduction programme carried out in conjunction with Midland Fertility Services, Aldridge, Birmingham, England. A pre-requisite for treatment was that on day 3 of the menstrual cycle the levels of follicle stimulating hormone (FSH) and oestradiol (E2) should be < 10 i mu/l and < 100 pg/ml respectively in the female partner. The ages of the women ranged from 26 to 42 years with a mean age of 35.5 years. Down regulation was carried out by using buserelin acetate 0.5 microgram subcutaneously from day 21 of the cycle for 21 days. This process was completed when the ovaries and pituitary gland were quiescent and the endometrial thickness < 4 mm in diameter. On completion of down regulation the gonadotrophin hormone, pergonal (dosage of 150-450 units) was used for ovarian hyperstimulation. A total of 294 oocytes (mean of 10.5, range 2-45) were retrieved of which 138 were fertilized (mean of 4.9, range of 0-28). Twenty-four patients each received a mean of two embryos. Five patients (20.8%) had positive pregnancy tests. Three patients (0.1%) developed ovarian hyperstimulation syndrome (OHSS), one had the severe, and two, the mild form of the syndrome. All three cases were treated successfully. The success at the initial IVF controlled ovarian hyperstimulation augers well for the future of infertile couples seeking treatment at the Fertility Management Unit, The University of the West Indies, Jamaica.

Adult↗

Optimizing hCG cut-off values: a single determination on day 14 or 15 is sufficient for a reliable prediction of pregnancy outcome.

OBJECTIVE: Optimizing the cut-off level for a single serum hCG determination around day 15 after oocyte retrieval or ovulation. STUDY DESIGN: Retrospective data analysis. RESULTS: 204 hCG samples >5 IU/L between March and October 1999 taken on day 14, 15 or 16 after oocyte retrieval in 204 patients undergoing IVF or ICSI were analyzed. ROC-curves and optimal cut-off levels to discriminate between viable and non-viable pregnancies were calculated for each day separately. Cut-off levels were found at 76, 142 and 223 IU/L for day 14, 15 and 16, respectively, and were verified by 487 hCG samples >5 IU/L taken between January 2000 and June 2004. CONCLUSIONS: A single serum hCG determination on day 14 or 15 is sufficient to discriminate viable pregnancies accurately from non-viable pregnancies.

Adult↗

Updated meta-analysis of recombinant follicle-stimulating hormone (FSH) versus urinary FSH for ovarian stimulation in assisted reproduction.

OBJECTIVE: To determine the relative efficacy of recombinant FSH (rFSH) and urinary FSH (uFSH) for ovarian stimulation in assisted reproductive techniques (ART). DESIGN: Systematic review and meta-analysis of randomized, controlled trials comparing rFSH and uFSH. SETTING: Infertility centers providing treatment with ART. PATIENT(S): Patients undergoing IVF with or without ICSI. INTERVENTION(S): Controlled ovarian stimulation using uFSH or rFSH (follitropin alpha or follitropin beta). MAIN OUTCOME MEASURE(S): Primary: rate of clinical pregnancy per cycle. Secondary: rates of spontaneous abortion, multiple pregnancy, and severe ovarian hyperstimulation syndrome (OHSS); total gonadotropin dose; serum E(2) level and number of follicles at hCG administration; number of oocytes retrieved. RESULT(S): Eighteen trials were included in the meta-analysis. Subgroup analyses demonstrated higher pregnancy rates with both follitropins compared with uFSH. However, statistical significance was reached only in the follitropin alpha versus uFSH comparison in IVF cycles, with an additional pregnancy for every 19 patients treated. Fewer units of rFSH than uFSH achieved the same E(2) level and oocyte yield. No differences were found between treatments in rates of spontaneous abortion, OHSS, and multiple gestation. CONCLUSION(S): rFSH produced higher pregnancy rates per cycle than uFSH when follitropin alpha was used in IVF, and the total gonadotropin dose required was lower.

Abortion, Spontaneous↗

The influences of weather on patients with different ovarian responses in the treatment of assisted reproductive technology.

PURPOSE: The purpose of this study was to evaluate the influences of tropical weather on patients with different ovarian responses in the treatment of assisted reproductive technology. METHODS: Six-hundred fourty-seven women underwent their first treatment cycles were retrospectively analyzed. Patients received embryo transfer either 3 days or 5 days after oocyte retrieval, depending on the number and quality of embryos on day-2. RESULTS: Significant correlations were demonstrated in the top quality embryo rates of day-3 and day-5 embryo transfers with temperature, humidity, and atmosphere pressure. The cumulative light hours negatively correlated with the implantation and pregnancy rates of day-3 embryo transfer (-.282 and -.282, respectively), while they positively correlated with those of day-5 embryo transfer (.225 and .224, respectively). CONCLUSIONS: These results clearly suggest that weather may exert influences on the outcome of assisted reproductive technology. Patients with different ovarian responses or blastocyst culture and transfer may modify weather influences.

Adult↗