JBRA Assist. Reprod. 2015; 19 (2):95-98
ORAL PRESENTATIONS
doi: 10.5935/1518-0557.20150022
Abstracts of the 12th RedLara Taller General, Lima, Peru, 26-29 March 2015
1Centro Nacional de Reproducción Asistida INNAIFEST
2Laboratorio de Biología Molecular de la Facultad de Medicina. Universidad Espíritu Santo
OBJECTIVE: Y chromosome microdeletions at the “Azoospermia Factor” regions (AZFa, AZFb, AZFc) are the second genetic cause of spermatogenic failure in infertile men. Despite its importance for the infertile patient treatment, any investigation about it has been published previously in Ecuador. The purpose of this study is to optimize a molecular technique that allows detection of microdeletions in the AZF region.
MATERIALS AND METHODS: Using a genomic DNA of healthy male with natural conceived offsprings, a multiplex real time polymerase chain reaction (qPCR) was standarized with eigth sequence-tagged site (STS) sY85, G34990, sY133, sY127, sY254, sY255, and using as internal control sex-determine region Y (SRY) and Ameologenin Y (AMELY). With this technique, 35 DNA samples taken from peripheral blood of patients with severe oligozoospermia were analyzed.
RESULTS: A triplex qPCR was standardized using EvaGreen DNA-binding dye to obtain melting temperature (Tm) of the STS previously mentioned. Three of the patients evaluated were detected to have partial microdeletion in the AZFa region, with a frequency of 8.8%; being losses in the G34990 section (one patient) and sY85 section (two patients). No cases of microdeletions in other AZF regions were found.
CONCLUSIONS: The triplex qPCR optimizated allows the identification of microdeletions in AZFa, AZFb and AZFc region in infertile men and a better clinical management of the patient’s treatment decision. This first report for Ecuador reveled a higuer prevalence of microdeletion in the AZFa region in comparison with those previously described in other populations.
1Private setting, International Center for Assisted Reproduction, Mendoza, Argentina
2Medical Director Immunology Laboratory, Central Hospital, Mendoza, Argentina
OBJECTIVE: To determine the role of polyvalent endometrial treatment in patients undergoing IVF-ET who had recurrent implantation failure (RIF) in a program of oocyte donation (OD). The results were expressed in terms of live birth rate (LBR). Secondly analyze changes of endometrial leukocyte population evaluated by flow cytometer (FC) and histopathology.
MATERIALS AND METHODS: Prospective study of a model-based control with analog abductive methodology. Over initial population of 75 patients with RIF in ovodonation, thirty cycles / patient of IVF/ET were selected in this study. A control group of 12 patients was established to variables FC. All patients were transferred to day 5-6 with a maximum of 2 expanded blastocysts with at least one of optimum quality. A versatile treatment was applied in all cases with both assessments in pre and postreatment.
RESULTS: Chronic endometritis was diagnosed in 14/30 (46.7%) with endometrial identifying germs in 12/30 (40%) and 6/30 (20%) was associated with endometrial thinning. A significant increase in endometrial thickness associated with a decrease in abnormal histopathology and Li/PC was observed at postreatment in relation with a pretreatment (P=0.047 and P=0.002) respectively. An increase of uterine killer cells (Nku) was observed in postreatment in absence of pregnancy. CD4/CD3 was established with prognostic value when their values are close to those of the control group.
CONCLUSIONS: Our findings demonstrate the reversibility of endometrial histological changes, both sonographics as immunological in RIF group under a polyvalent therapeutic; which is capable of modifying the immunology and endometrial histopathology and to obtain live birth.
1Clínica EUGIN, Bogotá, Colombia
2Clínica EUGIN, Barcelona 08029, Spain
3Fundació Privada EUGIN, Barcelona 08029, Spain
OBJECTIVE: A high body mass index (BMI) has been shown to associate with negative reproductive outcomes. Women with high BMI have in general lower chances of getting pregnant as well as higher risk of pregnancy complications. Several studies have described in the past the relationship between high BMI and the pregnancy outcome, however, some of them have a small sample size or fail to control for variables associated with a diminished probability of pregnancy. In the present study, we aim to analyze the role of the BMI of all parties involved in oocyte donation cycles (that is: the oocyte donor, the recipient woman, and the male partner) on pregnancy outcomes.
MATERIALS AND METHODS: This study includes 1092 oocyte donation cycles. Inclusion criteria were: fertilization by ICSI, frozen semen, transfer of 2 embryos at day 3 of in vitro development. For statistical analysis, BMI was divided in: low weight (<20 kg/m2), normal (20-24 kg/m2), overweight (25-29 kg/m2) and obesity (≥30 kg/m2). Quantitative and categorical variables were assessed by squared-Chi test and one-way ANOVA. The association between the BMI (recipient, oocyte donor and partner) and pregnancy rate was assessed by multivariate logistic regression.
RESULTS: Laboratory outcomes and pregnancy rates do not differ among the different BMI categories of recipient, oocyte donor or partner. After adjusted analyses (for oocyte donor age, for laboratory outcomes and for age and BMI of all the parties for pregnancy outcomes), no difference was found either.
CONCLUSIONS: In oocyte donation cycles, where donors BMI is by law mandated to be in the 18-30 range, the pregnancy rate of the oocyte recipient does not seem to be affected by the BMI of any of the parties involved.
1Centro de Estudios en Genética y Reproducción (CEGYR) Buenos Aires, Argentina
OBJECTIVE: To establish the relationship between oocyte cytoplasmic maturation and its chromosomal status and determine the effect of this feature over the reproductive outcome in patients with sub-optimal fertilization in ART.
MATERIALS AND METHODS: Fifty couples who underwent ART were selected. From nineteen patients, 22 metaphase II-MII and 18 failed-fertilized oocytes after ICSI were studied. The first polar body was collected for chromosomal analysis by aCGH. Oocytes were processed by immunocytochemistry-ICC to determine oocyte maturation: assessment of inactive MPF status and the conformation-alignment of the metaphase plate.
Other 31 couples presented sub-optimal fertilization (<50%) after ICSI, and failed-fertilized oocytes were studied by ICC. Two groups were conformed according to the main feature observed: A: cytoplasmic immaturity and sperm premature chromosome condensation and B: sperm nuclear decondensation failure with mature cytoplasm.
RESULTS: Regarding MII mature oocytes, 87% had a normal metaphase plate and 84% were chromosomally normal. Contrary, immature oocytes presented abnormal metaphase plate (86%) and just 33% were euploid.
In failed-fertilized oocytes: 100% of mature oocytes had a normal metaphase plate and 71% were euploid. When oocytes were cytoplasmic immature, 37% of them were normal (metaphase plate) and 50% were chromosomally normal. The global rate of aneuploidies and metaphase plate disarrangements in immature oocytes (MII+failed-fertilized) were significantly higher than mature oocytes (P<0.05). In patients with sub-optimal fertilization, the percentage of top quality embryos and pregnancy rate was significantly higher in group B (P<0.05).
CONCLUSIONS: Oocyte cytoplasmic immaturity is related to metaphase plate anomalies and aneuploidies. Fertilized oocytes, from a cohort with sub optimal fertilization with cytoplasmic immaturity, had poorer reproductive outcomes.
1Alciver Hospital – Fertility Unit Ecuador
OBJECTIVE: Determine the number of metaphase II oocytes that are needed for optimum rate blastulation and get a better rate of gestation.
MATERIALS AND METHODS: Retrospective study. Participants: Women diagnosed with infertility. Interventions: There were 110 women in which they are performed IVF-ICSI, excluding 3rd day transference and transfer canceled cycles. After controlled ovarian stimulation, cultivation continues to blastocysts. Main outcome measures: Rate of metaphase II oocytes, pregnancy rate. Average age of patients, IVF-ICSI technique.
RESULTS: Mean age of the patients 33.41y/o. IVF technique: 73 patients, 66.4%; ICSI: 37 patients 33.6%. Pregnancy: Yes: 57.3%; No: 42.7%. Number of oocytes in metaphase II: Average of 7.5.
CONCLUSIONS: The pregnancy rate coincides with the center’s results (50-60%), reach from 7 mature oocytes. The blastulation average rate was 39.2%.
1Centro de Fertilidad y Ginecología del Sur, Cusco, Peru
OBJECTIVE: Determine the effectiveness of in vitro maturation of oocytes in the infertility treatment in high altitude women with polycystic ovaries.
MATERIALS AND METHODS: Design: descriptive and retrospective study. Participants: Women with polycystic ovaries and infertility. Interventions: there were 11 women from locations above 7,546 feet above sea level with polycystic ovaries and infertility in which were performed in vitro maturation of oocytes, followed by intracytoplasmic sperm injection, culture and embryo vitrification. After that, the endometrium was prepared and the embryos were thawed and transferred. Main results mesurements: oocytes maturation, fecundation, clinical pregnancy and implantation rates.
RESULTS: oocytes maturation rate was 86.1%; fecundation rate 90.3%; clinical pregnancy rate 36.4% and implantation rate 17.4%.
CONCLUSIONS: In vitro maturation of oocytes is an effective technique in the infertility treatment of high altitude women with polycystic ovaries.
1CAPSIR- Argentine Center of Psychology and Reproduction
INTRODUCTION: We propose in this paper a brief road of some aspects that affect people with infertility problems and future children.
REVIEW: Making the decision to have a child and accept medical aid represents an important in the life of a couple, or persons, with problems in procreation step. Then when the baby is born, begins a common path to all parents, until the question that every child gets done, “How was I born?” Or “How babies are born?”. In the case of a man and a woman, who resorted to reproductive medicine, the answer requires a particular approach, often revives internal conflicts and so often postponed. All children need to know their history of origin, and at some point begin to investigate how they were born. That question, often disturb most parents, it becomes more difficult to answer in the case of assisted reproduction, because many times parents have not resolved the issue. Still, parents who from the beginning wanted to share with your child the story of her conception, may find it difficult to imagine how. In other cases, moreover, is not so easy to share the fact that medical help is required and sometimes the donation of gametes (eggs or sperm) to reach the desired pregnancy. The point is to think and talk about this with the children, because children ask to the extent that parents give space for questions and dialogue. What is noteworthy is the right of children to receive that knowledge. From psychology, we know that the ‘secrets’ family who do not communicate to children can cause emotional disorders, various symptoms in both children and parents, and cause restlessness and anxiety.
COMMENTS: Our experience indicates that some children with behavioral problems improved significantly after his family spoke about his origin. Children need us to communicate with truths. They, then develop their own theories and fantasies, but with a real base. What we can not do is lie or remain silent.
1Centro de Fertilidad y Reproduccion Asistida (CEFRA), Lima- Peru
REDLARA AWARD Best Oral Presentation
12º REDLARA General Congress - Lima - Peru 2015
OBJECTIVE: To determine the effect of using MACS technology on clinical pregnancy, as a method for separation of damaged sperm in infertile patients.
MATERIALS AND METHODS: 136 infertile men having normal semen parameters in accordance with WHO 2010 criterion, undergoing ICSI cycle were enrolled during the course of the study. The patients were prospectively randomized and enrolled after oocyte retrieval and were assigned to the ICSI group, PICSI group or MACS group. Embryo development and clinical pregnancy were assessed. In 17 randomized MACS patients, sperm DNA fragmentation was tested in the presumptive apoptotic and no apoptotic spermatozoa fractions.
RESULTS: Similar results were obtained between groups for the following parameters: fertilization rates of 78.97% (95% confidence interval [CI], 74.37 - 83.57), 70.15 %(95% CI, 63.98 - 76.33) and 80.28% (95% CI, 73.74 - 86.81) for ICSI, PICSI and MACS group, respectively; Number of Day-3 embryos was 5.04 (95% CI, 4.09 - 5.98), 5.17 (95% CI, 4.24 - 6.10) and 5.59 (95% CI, 4.31 - 6.87) for ICSI, PICSI and MACS group, respectively; number of freezing embryos in blastocyst stage was 0.78 (95% CI, 0.25 - 1.31), 0.70 ( 95% CI, 0.27 - 1.14) and 1 (95% CI, 0.37 - 1.6) for ICSI, PICSI and MACS group, respectively. However, clinical pregnancy rates of 58.1% for MACS group versus 40.4% and 27.3% for PICSI and ICSI group, respectively, were showed statistical difference (P= 0.019). DNA fragmentation index for the two sperm MACS fraction showed statistical differences (P= 0.000); MACS reduced the D.F.I of the sperm sample.
CONCLUSIONS: The use of MACS technology improves the clinical pregnancy on infertile couples and can be applied as a method for sperm separation, discriminating sperm with high DNA fragmentation.
1HISPAREP Fertility Clinic, Hospital Español, México DF
2Multidisciplinary Diabetes Center México City (Hyperbaric Chamber)
OBJECTIVE: To assess whether hyperbaric oxygen sessions elevate serum levels of anti-Müllerian hormone (AMH) in patients diagnosed with infertility with serum levels of less than or equal to 1 ng/dl AMH.
MATERIAL AND METHODS: A study was performed on 4 patients diagnosed with infertility. Serum AMH level was measured at the beginning and end of hyperbaric oxygen sessions, and endometrial thickness was measured on endometrial cycle day 14 before and during the hyperbaric oxygen sessions.
RESULTS: In two of the four patients, the serum AMH level increased by 40% and 116%. In one patient the serum AMH level was not elevated, with a serum AMH level before and after treatment of 0.1 ng/dl. The fourth patient became pregnant during the hyperbaric oxygen sessions. Endometrial thickness was not improved in any of our patients.
CONCLUSIONS: This study showed that hyperbaric oxygen sessions can increase serum AMH levels, with a significant increase of 116% in one case.
Therefore, this therapy can be used as an alternative treatment for patients with serum AMH levels of less than or equal to 1 ng/dl and a limited number of eggs for IVF cycles but not for patients with serum AMH levels of less than or equal to 0.1 ng/dl, as we did not observe an increase in serum AMH level in patients with an initial AMH level of 0.1 ng/dl. This study did not demonstrate improvement in endometrial growth following hyperbaric oxygen sessions.
1Centro de Estudios en Genética y Reproducción (CEGYR)
Buenos Aires, Argentina
OBJECTIVE: The aim of the present study is to assess the correlation between the presence, quantity and size of nuclear vacuoles and DNA damage and chromatin status in sperm samples of men who underwent to assisted reproduction technology.
MATERIAL AND METHODS: Forty six males who underwent to assisted reproductive technology (ART) were considered. According to their latest semen analysis (<3 months), were grouped into: (A) strict morphology index ≤4% (26) and (B) strict morphology index ≥14% (20). Motile sperm were selected by density gradient, and MSOME study was conducted to assess the number and size of nuclear vacuoles. DNA fragmentation (TUNEL) and DNA strand status (acridine orange) were assessed over the selected spermatozoa accordingly to their vacuole pattern.
RESULTS: In group A, sperm without vacuoles (1) have similar levels of DNA fragmentation (TUNEL) in compare to the rest of observed patterns (2°- 6°). Regarding to AO, spermatozoa with large or several vacuoles that cover more than 30-50% of the nuclear surface are AO+, but not necessarily TUNEL positive. The first three patterns of vacuoles patterns had lower levels of AO in compare to grades 4° and 6°. In group B, those sperm with one or more vacuoles greater than 30%-50% (4° and 6°), had a significant increase in TUNEL values, in relation to group 1°- 3°. Considering AO, it was found that the 4° and 6° pattern had a significantly elevated level of this marker, as same of group A (P <0.05).
CONCLUSIONS: There is no relationship between the greater number and size of sperm vacuoles with high levels of DNA fragmentation in patients with severe teratozoospermia (Krueger <4%). Conversely, this relationship is evident in normal semen samples (normal morphology).
Sperm selection by IMSI technique, to select non-fragmented sperm in patients with Krueger <4%, is not necessarily secured when non-vacuolated sperm is selected.
1FERTIPRAXIS – Reproducao Humana, Rio de Janeiro, RJ, Brasil
2Instituto Nacional de Cancer (INCA), Rio de Janeiro, RJ, Brasil
INTRODUCTION: Cancer survivors are increasing and assisted reproductive technologies (ART) developments are also more common. So, fertility preservation of the reproductive-age women with cancer is emerging as a challenging but rewarding application of ART. Ovarian tumors diagnosed in young ages tend to be low-stage low-grade malignancies. Although fertility saving surgery is more often confined to early-stage and low-grade disease, it also has been reported in advanced stages (up to Stage IIIc). Management of borderline ovarian tumors has evolved significantly in the last few decades and in contrast to invasive ovarian cancer, they can be operated on conservatively at all stages.
CASE REPORT: A left borderline serous ovarian tumor stage I (FIGO) was discovered and treated in a 26-year-old black nulliparous woman, by conservative approach (laparotomy, salpingo-oophorectomy). In a six months interval she had a 7.5 x 7.1 x 5.7cm multilocular contralateral tumor with septa and vegetative areas and in a year interval a CT showed a 8.4 x 7.4 x 7.0 lesion that precluded a follicular aspiration.
The authors discuss the multidisciplinary strategy and the approach with the couple: the best option considered would be the resection of the tumor remaining attached to a new study pelvic. If possible uterine conservation should be held for further procedure of oocyte donation. Two years from the first surgery she had the second laparotomy and six months later she had an ICSI with oocyte donation.
She became pregnant and delivered two 34 week-pregnancy boys through C-section due to hypertension plus preeclampsia. There happened a post-operative intestinal obstruction that required a new surgical approach to adhesions lysis. There was no report of tumor lesions then.
Seven months later, mother and children are doing well. Comments are made about borderline ovarian tumors and fertility-sparing approaches.
COMMENTS: Our report serves to confirm that fertility-sparing approaches are well tolerated not only in patients with early-stage Borderline ovarian tumors (BOTs) but also in patients with advanced-stage BOTs with noninvasive extraovarian implants, if these implants can be resected completely.
1Unidad de Fertilidad UNIFERTES, Caracas, Venezuela
OBJECTIVE: 1. To describe the standardization process and protocols of the ET method at our center.
2. To compare the performance of non-echogenic catheters with echogenic catheters during ultrasound-guided ET.
MATERIAL AND METHODS: Retrospective analysis of 2630 ET performed at UNIFERTES during 1997-2014, to describe standardization process and to compare the percentage of difficult ET between echogenic and non-echogenic catheters. We tested 17 non-echogenic and three echogenic catheters.
RESULTS: Many variables were associated with the ease of ET : informed patients, waiting time for the procedure, speculum use, clinical touch, uterine contractions, cervical mucus removal, presence of blood before or after the procedure, full bladder, ultrasound guidance, uterocervical angle, mock transfer, catheter type (soft or hard, echogenic or non-echogenic, with stylet or not), catheter loading technique, duration of embryo loading (time interval since the embryos were removed from the incubator for loading until the catheter is passed to the physician), transfer procedure (time interval from the catheter was handed to the physician until the embryos were discharged in the uterus), catheter tip placement, retained embryos, bed rest after ET, operators proficiency.
The diversity of catheters used and the percentage of difficult transfers decrease as the use of echogenic catheters increases. This process is necessary to minimize variation, ensure high quality, safe and evidence-based practice, and improve outcomes.
CONCLUSIONS: To standardize the ET method allowed a quicker and easier transfer. The use of echogenic catheters simplified ET procedures guided by abdominal ultrasound.