P.N. Barri

Screening for abnormalities of chromosomes X, Y, and 18 and for diploidy in spermatozoa from infertile men participating in an in vitro fertilization-intracytoplasmic sperm injection program

OBJECTIVE To evaluate the frequency of disomy (for chromosomes X, Y, and 18) and of diploidy in the spermatozoa of infertile men undergoing intracytoplasmic sperm injection (ICSI). DESIGN Prospective analysis of sperm nuclei by fluorescence in situ hybridization (FISH). SETTING University-affiliated IVF-ICSI program. PATIENT(S) Semen samples from 19 patients participating in an IVF-ICSI program. INTERVENTION(S) Semen samples were analyzed and prepared for FISH. MAIN OUTCOME MEASURE(S) Semen parameters were evaluated. The frequency of disomy for chromosomes X, Y, and 18 and the frequency of diploidy were analyzed by FISH. RESULT(S) A total of 9,373 spermatozoa from 19 infertile patients were analyzed and compared with spermatozoa from a control group of 5 healthy men. No differences in the frequency of disomy 18 were found, but statistically significant differences in the incidence of sex chromosome disomy and of diploidy were observed. CONCLUSION(S) The study of sperm nuclei by FISH is useful to improve genetic counseling in infertile patients selected for ICSI.

Laser blastocyst biopsy for preimplantation diagnosis in the human

A new methodology for blastocyst biopsy that uses a 1.48 microm diode laser is described. Trophectoderm cells are biopsied after laster zona drilling and culture, fixed and processed for fluorescent in situ hybridisation (FISH) analysis. Preliminary results on the efficiency of the procedure and blastocyst recovery rate are promising. Blastocyst laser biopsy is a useful tool in preimplantation genetic diagnosis (PGD) as it allows a more reliable diagnosis and widens the diagnostic possibilities on account of the higher number of cells obtained in the biopsy.

Confirmation of diagnosis in preimplantation genetic diagnosis (PGD) through blastocyst culture: preliminary experience.

Three cases of preimplantation genetic diagnosis (PGD) (two for sexing and one for aneuploidy screening) are presented. Embryo biopsy was performed at day 3 and diagnosis was established with fluorescent in situ hybridization (FISH). Embryos not used for replacement were cultured in sequential media for blastocyst development. Blastocyst rate was 39.3 per cent. Confirmations of diagnosis were established with FISH in blastocysts and arrested embryos. Mosaicism was observed in 7/8 blastocysts (mean number of cells analysed: 55.5) and 5/8 arrested embryos. The percentage of abnormal cells was 17.1 per cent for blastocysts and 54 per cent for arrested embryos. Polyploid cells were observed in 4/8 blastocysts. Confirmation of diagnosis at the blastocyst stage is a useful tool in PGD. Copyright

Increased Incidence of Meiotic Anomalies in Oligoasthenozoospermic Males Preselected for Intracytoplasmic Sperm Injection

AbstractPurpose: Based on data from the literature, to detect thepossible presence of an increased frequency of meiotic anomaliesin oligoasthenozoospermic (OA) patients preselectedfor intracytoplasmic sperm injection. Methods: Meiotic studies in as many successive patientswith a clinical indication for a diagnostic testicular biopsyas needed to complete at least 100 cases with a severe OA(motile sperm concentration ≤1.5 × 106/ml). Results: An increased incidence of meiotic anomalies wasfound in 102 patients with a severe OA (17.6%) comparedto the mean for 105 patients with other etiologies in theseries (5.7%) or the mean for patients reviewed in the literature(6.5%). Conclusions: Patients with a severe OA have a higher incidenceof synaptic anomalies. This may result in themalsegregation of chromosomes at meiosis I, producingabnormal sperm, and could explain the high incidence ofsterility and some cases of abortion (in two thirds of thecouples with abortions the husband had meiotic anomalies)in this group.

Human chorionic gonadotropin and intravaginal natural progesterone are equally effective for luteal phase support in IVF

This prospective randomized study compared human chorionic gonadotropin (hCG) and micronized transvaginal progesterone for luteal support in 310 in vitro fertilization (IVF) patients treated with leuprolide acetate and gonadotropins in a long protocol, and showing normal ovarian response. Both treatment groups were homogeneous for age, BMI, stimulation treatment and ovarian response. Pregnancy rates per embryo transfer were not significantly different (33.1% for the hCG group versus 38.7% for the progesterone group). For IVF patients with a normal response to stimulation under pituitary supression, the use of hCG or progesterone for luteal support does not seem to have any effect on pregnancy rate. The choice of luteal treatment must balance medical hazard and patient convenience, as both therapeutic regimens seem equally effective.

Analysis of factors associated with multiple pregnancy in an oocyte donation programme

The aim of this study is to identify the factors associated with multiple pregnancy in an oocyte donation programme. A retrospective study (2000-2007) of 945 synchronous cycles was performed. Two embryos were transferred in all cycles on day 2 after oocyte retrieval. All variables (egg donor and recipient age, number of inseminated oocytes, fertilized oocytes, cleaved embryos, good-quality embryos available, good-quality embryos transferred and frozen embryos) were analysed in relation to the clinical pregnancy rate per transfer (PR) and the multiple pregnancy rate (MPR). The donor age was 26.8±4.5 years and recipient age was 41.0±5.4. The number of good-quality embryos per recipient was 3.1±2.5. The PR was 55.1% and the MPR 36.5%. The number of good-quality embryos transferred (2 versus 0) was significantly associated (P<0.05) with the PR (60.6% versus 43.5%). The relationship between the MPR and the number of good-quality embryos transferred was adjusted by donor and recipients age. For those patients who received 2 versus 0 good-quality embryos, the odds ratio of a multiple pregnancy was 2.1 (95% CI 1.121-3.876). The only predictive factor for multiple pregnancies in an oocyte donation programme is the quality of the transferred embryos. Since the development of assisted reproduction techniques, most countries have witnessed increased rates of multiple pregnancy. In IVF/intracytoplasmic sperm injection, effective strategies must be established to prevent multiple pregnancy without reducing overall pregnancy rates. In oocyte donation programmes, there is less awareness about the related risks. Actually, a minimum of two embryos are still transferred in most centres. The patients age is higher and this fact implies more obstetric and perinatal complications. The aim of this study is to identify the factors associated with multiple pregnancy to contribute to establish future guidelines to avoid those risks.

Birth after transfer of frozen-thawed vitrified biopsied blastocysts

Purpose: To present a case describing the birth of a healthy female after the replacement of vitrified biopsied embryos after Preimplantation Genetic Diagnosis.Method: A descriptive case report of a single patient.Results: Our patient carrier of an X-linked disease became pregnant and as a result a healthy girl was born.Conclusions: This report shows that blastocysts obtained from biopsied embryos can be successfully cryopreserved by a simple, secure and low-cost vitrification method using a Hemi-straw support.

Preimplantation genetic diagnosis in patients with male meiotic abnormalities.

Indications and candidates for preimplantation genetic diagnosis (PGD) have increased in recent years. This study evaluates whether IVF-intracytoplasmic sperm injection (ICSI) results could be improved by selecting embryos through PGD-AS (aneuploidy screening) in couples in whom the male partner presents meiotic abnormalities. Two hundred and fifty-six embryos were biopsied and 183 were suitable for analysis (73.2%). Ninety-two embryos showed normal chromosomal analysis (50.3% of the analysed embryos and 57.5% of the diagnosed embryos). Pregnancy, abortion and implantation rates were compared with 66 IVF-ICSI cycles performed in 44 patients with meiotic abnormalities without PGD (control group). No statistically significant differences in the pregnancy rate (52 versus 43.9%), implantation rate (32.1 versus 23.5%) and miscarriage rate (15.4 versus 10.3%) were observed between the groups. Although the embryos obtained from men with meiotic abnormalities showed a high frequency of chromosome abnormalities, no improvements in pregnancy and implantation rates were obtained after PGD-AS in the series analysed.

Correlation between embryological factors and pregnancy rate: development of an embryo score in a cryopreservation programme

PurposeTo establish which embryo parameters, in frozen thawed embryo transfers, have the highest prognosis value in the establishment of pregnancy. The relative importance of different embryo parameters is used to develop an embryo score.MethodsRetrospective analysis of the implantation rate in 356 frozen/thawed single embryo transfers. A logistic regression model is used to establish an embryo score.ResultsA direct correlation is established between the implantation rate and fresh embryo development (number of blastomeres and their symmetry), survival rate after thawing and mitosis resumption after overnight culture.ConclusionsAn embryo score is developed to determine the implantation potential of frozen/thawed embryos.

Combined strategy for fertility preservation in an oncologic patient: vitrification of in vitro matured oocytes and ovarian tissue freezing

Breast cancer is the most common malignancy in women and nearly 7% of cases are diagnosed before age 40 [1]. Diagnosis at early ages is associated with a worse prognosis and therefore, these cases are usually managed with adjuvant chemotherapy and/or hormonal therapy. These treatments are effective in reducing recurrence rates and improving survival rates; however, they have a negative impact on ovarian function and fertility [2]. Fertility preservation in these cases may be of interest also due to the current trend towards delaying motherhood, which leads to a situation where many women may have not fulfilled their motherhood desire at the time of breast cancer diagnosis. There are several strategies to preserve fertility. Choosing one or the other will depend on patient’s age, tumour biology and, above all, time availability until the start of gonadotoxic treatment.