JBRA Assist. Reprod. 2017;21(2):137-141
ORAL PRESENTATIONS
doi: 10.5935/1518-0557.20170030
Abstracts of the 13th Red Lara Taller General, Buenos Aires, Argentina, 26-28 April 2017
E. Borges Jr.1,2, D.P.A.F. Braga1,3, B.F. Zanetti1, A.S. Setti2, R.R. Provenza1, A. Iaconelli Jr.1
1Fertility - Medical Group, São Paulo, SP - Brazil
2Instituto Sapientiae - Centro de Estudos e Pesquisa em Reprodução Humana Assistida, São Paulo, SP - Brazil
3Disciplina de Urologia, Área de Reprodução Humana, Departamento de Cirurgia, Universidade Federal de São Paulo. - UNIFESP
Objective: To discuss the requirement of the National Health Surveillance Agency (ANVISA), to assisted reproduction treatment patients undergo laboratory tests for ZIKV detection, and also if it's prudent the public health authorities and government leaders recommendation to women simply avoid pregnancy.
Methods: This study was performed in a university-affiliated In vitro fertilization center in Brazil. A critical discussion on the risk of microcephaly due to ZIKV infection and prevalence of others harmful pathogens to vulnerable pregnant women and infants was conducted. Moreover, 954 patients undergoing intracytoplasmic sperm injection cycles (ICSI), between April and November of 2016, were evaluated concerning the results of ZIKV test, according to the region of Brazil.
Results: Patients undergoing ICSI cycles were split into groups according with region origin: 28 (3.0%) were from the North, 27 (2.8%) were from the Northeast, 40 (4.2%) were from the Central West, 830 (87.2%) were from Southeast, and 29 (3.0%) were from the South. Concerning the diagnosis, 112 samples had a positive or inconclusive result for ZIKV by chromatography immunoassay. These samples were re-analyzed by ELISA and none result was positive. All positive results were from the Southeast region and none from the Northeast or Central West regions, which are considered endemic regions.
Conclusion: ZIKV test before the beginning of assisted reproduction treatments does not exclude the risk of the infection during pregnancy. In addition, although ZIKV infection risk is extremely high, the microcephaly risk due to ZIKV is not higher than the risk of miscarriage and birth defects due to other recognized pathogens.
E. Borges Jr.1,2, B.F. Zanetti2, A.S. Setti2, D.P.A.F. Braga1,3, R.C.S. Figueira1, A. Iaconelli Jr.1
1Fertility - Medical Group, São Paulo, SP - Brazil
2Instituto Sapientiae - Centro de Estudos e Pesquisa em Reprodução Humana Assistida, São Paulo, SP - Brazil
3Disciplina de Urologia, Área de Reprodução Humana, Departamento de Cirurgia, Universidade Federal de São Paulo. - UNIFESP
Objective: To determine the effect of FSH doses on intracytoplasmic sperm injection (ICSI) outcomes according to the age of the patient.
Methods: Patients undergoing controlled ovarian stimulation (COS) for ICSI cycles in a university-affiliated in vitro fertilization center were split into age groups: ≤35 y.o. (n = 1523); >35 and ≤38 y.o. (n=652); >38 and ≤40 y.o. (n = 332); and >40 y.o. (n = 370). The effect of FSH dose on COS, laboratorial and clinical outomes was determined by linear regression models.
Results: The FSH dose didn't affect the ovarian response in terms of total number of follicles, retrieved oocytes and mature oocytes within the age groups, but it was observed that the lower the age, the lower the FSH dose is needed per oocyte retrieved. In the group of patients ≤35 y.o., it was also noted a positive effect of FSH dose on oocyte yield. Despite that, for patients ≤38 y.o. it was verified a negative effect of FSH dose on embryo quality and blastocyst formation rate, and also an increase in cycle's cancelation rate. In patients ≥ 39 y.o., there were no effects of the FSH doses on the analysed variables.
Conclusion: The ovarian stimulation with high doses of FSH is not recommended in younger women (≤38 y.o.), once it was observed a decrease in embryo quality and an increase in cycle's cancelation rate. Mild ovarian stimulation protocols may be more appropriate; however it may not be applicable for women in advanced age, since its necessary a higher FSH dose for oocyte retrieval in these patients.
A.V. Sampo1, C. Palena1, L. Ganzer1, V. Maccari1, G. Estofán1, M. Hernández1
1CIGOR - Centro Integral de Ginecología, Obstetricia y Reproducción. Córdoba, Argentina
Objective: To assess the effect of Body Mass Index (BMI) on the results obtained in ICSI cycles.
Methods: We studied 266 ICSI cycles realized between January 2014 and December 2016. Patients were grouped according to their BMI in: Normal (18.5-24.9), Overweight (25.0-29.9) and Obese (>30). We compared between the groups: antral follicles number, ovarian stimulation lenght, gonadotropin dose used, maximum oestradiol value, follicles developed /antral follicles, retrieved oocytes/developed follicles and mature/retrieved oocytes, normal fertilization rate, embryo achieved/normal fertilized oocytes, clinical pregnancy and implantation rate. Kruskal-Wallis and Chi square test were used. P<0.05 was considered significant.
Results: Normal, Overweight and Obese patients presented comparables values for antral follicles number (11.6±5.4, 12.5±5.5, 12.2±5.7), ovarian stimulation length (7.5±1.4, 7.6±1.1, 7.8±1.3) and gonadotropin dose used (2043±489, 1940±536, 2109±605). Obese patients had lower values of oestradiol (1560±610, 1511±635, 1190±466; P=0.018), developed follicles (81%, 76%, 70%; P<0.0001); and retrieved oocytes (91%, 90%, 84%; P=0.0017); and not significantly lower values of mature oocytes (82%, 82%, 77%; P=0.26). Groups had comparable fertilization rate (72%, 73%, 69%) and embryo achieved rate (67%, 63%, 72%). Normal group showed higher but not significant pregnancy and implantation rates (43%, 40%, 38%, P=0.53; and 33%, 26%, 23%; P=0.11), and significantly higher ongoing pregnancy rate (37%, 33%, 33%, P=0.042)
Conclusion: Increased BMI patient had impaired ovarian response and pregnancy rate in ICSI cycles.
M.C.A. Cardoso1, A. Evangelista1,2, C. Sartório1, G. Vaz1,2, C.L.V. Werneck1, F.M. Guimarães1, P.G. de Sá1,2, M.C. Erthal1
1Vida - Centro de Fertilidade, Rio de Janeiro, Brazil
2Department of Gynecology of the State University of Rio de Janeiro (UERJ)
Objetive: Evaluate the efficacy of the double-stimulation protocol over conventional ovarian stimulation in order to recover a more adequate number of oocytes to increase the number of embryos to be transferred or to be genetically analyzed.
Methods: A retrospective and comparative study with 13 patients who underwent unsuccessful IVF cycles with a conventional antagonist ovarian stimulation protocol and repeated the attempt with a double stimulation protocol. The following variables were analyzed: number of oocytes collected, mature oocytes collected, fertilization rate, blastocyst rate, biopsied blastocyst rate and euploidy rate.
Results: The double stimulation protocol had a significant higher number of oocytes collected (P=0.007) and mature oocytes to be injected (P=0.01). There was no statistical difference in fertilization (P=0.78) and blastocyst (P=0.59) rates.
Conclusion: Double stimulation favors patients who are at risk of incurring several attempts of IVF in order to achieve pregnancy.
J.G. Franco Jr1,2, C.G. Petersen1,2, A.L. Mauri1,2, L.D. Vagnini2, A. Renzi2, B. Petersen2, M.C. Mattila1, V.A. Comar1, J. Ricci1, F. Dieamant1,2, J.B.A. Oliveira1,2, R.L.R. Baruffi1,2
1Center for Human Reproduction Prof. Franco Jr., Ribeirao Preto, Brazil
2Paulista Center for Diagnosis, Research and Training, Ribeirao Preto, Brazil
Objective: KPIs have been employed for internal quality control (IQC) in ART. However, clinical KPIs (C-KPIs) such as age, AMH and number of oocytes collected are never added to laboratory KPIs (L-KPIs), such as fertilization rate and morphological quality of the embryos for analysis, even though the final endpoint is the evaluation of clinical pregnancy rates. This paper analyzed if a KPIs-score strategy with clinical and laboratorial parameters could be used to establish benchmarks for IQC in ART cycles.
Methods: In this prospective cohort study, 280 patients (36.4±4.3years) underwent ART. The total KPIs-score was obtained by the analysis of age, AMH (AMH Gen II ELISA/pre-mixing modified, Beckman Coulter Inc.), number of metaphase-II oocytes, fertilization rates and morphological quality of the embryonic lot.
Results: The total KPIs-score (C-KPIs + L-KPIs) was correlated with the presence or absence of clinical pregnancy. The relationship between the C-KPIs and L-KPIs scores was analyzed to establish quality standards, to increase the performance of clinical and laboratorial processes in ART. The logistic regression model (LRM), with respect to pregnancy and total KPIs-score (280 patients/102 clinical pregnancies), yielded an odds ratio of 1.24 (95%CI = 1.16-1.32). There was also a significant difference (P<0.0001) with respect to the total KPIs-score mean value between the group of patients with clinical pregnancies (total KPIs-score=20.4±3.7) and the group without clinical pregnancies (total KPIs-score=15.9±5). Clinical pregnancy probabilities (CPP) can be obtained using the LRM (prediction key) with the total KPIs-score as a predictor variable. The mean C-KPIs and L-KPIs scores obtained in the pregnancy group were 11.9±2.9 and 8.5±1.7, respectively. Routinely, in all cases where the C-KPIs score was ≥9, after the procedure, the L-KPIs score obtained was ≤6, a revision of the laboratory procedure was performed to assess quality standards.
Conclusion: This total KPIs-score could set up benchmarks for clinical pregnancy. Moreover, IQC can use C-KPIs and L-KPIs scores to detect problems in the clinical-laboratorial interface.
F.P. Déniz1, C. Encinas1, J. La Fuente1
1Embriovid, La Paz, Bolivia
Objective: To describe a patient selection methodology for elective single embryo transfer (eSET) eligibility, emphasizing the inclusion criteria and their results.
Methods: Retrospective analysis. All cases carried out in a private clinic between June 2011 and December 2016, in La Paz, Bolivia (3600 meters above sea level). In 34 IVF/ICSI cycles, elective single embryo transfer was performed, all of them in blastocyst stage. Gardner's blastocyst classification criteria were used. Between the two stages of the study (July 2015), each embryo grade implantation rate was recalculated, as a consequence, the inclusion criteria were expanded.
Results: The clinical pregnancy rate of the 34 cases in the first transfer was 55.9% (19/34). Neither twin nor multiple pregnancies occurred. To date, the cumulative pregnancy rate is 64% [(19+3)/34]. The first stage comprised 2.56% (12/468) of patients who accessed the elective single embryo transfer program; the implantation rate was 58.3% (7/12). Whereas in the second stage, 14.29% (22/154) of patients were eligible, the implantation rate was 54.55 (12/22) respectively.
Conclusion: The implementation of an eSET programme based on an in-depth morphological embryo assessment combined with the calculation of the implantation potential of each embryo grade are a suitable method to achieve acceptable clinical outcomes and to reduce multiple pregnancies in patients to whom two embryos are transferred. It is mandatory that every clinic must be aware of the implantation rate of each embryo grade in its own setting.
T. Puga-Torres1, X. Blum-Rojas1, M. Blum-Narváez1
1Centro Nacional de Reproducciónn Asistida INNAIFEST - Guayaquil - Ecuador
Objetive: Identify different blastocyst classification systems that are used by embryologists in Latin American countries and evaluate the possibility of establishing a consensus among these countries.
Methods: An email survey was conducted through the Latin American Network of Assisted Reproduction (REDLARA) aimed at embryologists of assisted reproduction centers in Latin countries.
Results: Sixty surveys were collected from 12 Latin American countries, of which 66.7% had >10 years of professional practice as embryologists. Seven different blastocyst classification systems were reported, of which 5 have previously been described in the literature.
Conclusion: Although the group of embryologists surveyed use different blastocyst classification systems, the majority of this group considers that the embryo score system should be unified in their countries as well as in the region.
V.A. Comar1, C.G. Petersen1,2, A.L. Mauri1,2, M.Mattila1, L.D. Vagnini2, A. Renzi2, B. Petersen2, A. Nicoletti1, F. Dieamant1,2, J.B.A. Oliveira1,2, R.L. R. Baruffi1,2, J.G. Franco Jr.1,2
1Center for Human Reproduction Prof. Franco Jr., Ribeirao Preto, SP, Brazil
2Paulista Center for Diagnosis Research and Training, Ribeirao Preto, SP, Brazil
Objective: The aim of this study was to evaluate the influence of different periods of abstinence on conventional seminal parameters as well as functional parameters in human semen, including mitochondrial function, chromatin packing and sperm DNA fragmentation.
Methods: A cohort of 2,458 men undergoing infertility investigation was recruited. Semen analyses were performed according to WHO guidelines/morphology-motile sperm organelle morphology examination/MSOME. For DNA integrity analysis, the percentages of DNA fragmentation (TUNEL), abnormal chromatin packaging/underprotamination (chromomycin A3/CMA3), abnormal mitochondrial membrane potential (MMP/MitoTracker Green), and apoptosis (annexin-V) were recorded. Associations between the sexual abstinence period and sperm parameters were assessed using Spearman correlation. For group comparisons, the subjects were categorized according to the sexual abstinence period (SAP) into three groups: SAP <2 days, SAP 2-5 days, and SAP >5 days.
Results: The duration of abstinence had a statistically significant positive influence on sperm concentration, volume, and the number of leukocytes and a statistically significant negative influence on sperm motility and vitality. The percentages of DNA fragmentation and MMP(mitochondrial damage) worsened with the increased duration of abstinence. The percentage of sperm protamination was statistically significantly increased with abstinence.
Conclusion: Increase in the sexual abstinence period influences sperm quality. This study reinforces the importance of the duration of ejaculatory abstinence on semen parameter variation. It highlights the deleterious effect of increased abstinence on DNA damage, which is most likely associated with ROS (mitochondrial damage/number of leukocytes). The increase in chromatin packaging can represent a protective feature for DNA.
F. Dieamant1,2, C.G. Petersen1,2, A.L. Mauri1,2, V.A. Conmar1, M.C. Matilla1, L.D. Vagnini2, A. Renzi2, B. Petersen2, C. Zamara1, J.B. A. Oliveira1,2, R.L.R. Baruffi1,2, J.G. Franco Jr.1,2
1Centre for Human Reproduction Prof Franco Jr, Research, Ribeirao Preto, Brazil
2Paulista Center for Diagnosis Research and Training, Research, Ribeirao Preto, Brazil
Objective: To evaluate the effects of varicocele on the conventional semen parameters: sperm DNA fragmentation, chromatin packaging, mitochondrial membrane potential (MMP) and sperm apoptosis.
Methods: A cross-sectional study was conducted with semen samples from 2,399 men from couples who attended an infertility clinic. A total of 16.3% (391/2399) of the men were diagnosed with varicocele by a urologist.
Results: Regression analysis revealed that the percentages of sperm presenting with DNA fragmentation, abnormal chromatin packaging, and abnormal MMP were significantly increased in individuals with varicocele compared to men without varicocele. Apoptosis was not influenced by varicocele. Conventional semen parameters were significantly worse in individuals with the pathology. On the other hand, in men with varicocele, Spearman's correlation demonstrated that early apoptosis and abnormal MMP showed a positive and significant correlation with sperm DNA fragmentation.
Conclusion: Men with varicocele had worse conventional semen parameters, including increased levels of sperm DNA fragmentation, inactive mitochondria, and abnormal chromatin packaging. These alterations are possible causes of infertility in individuals with varicocele.
C. Alvarez Sedó1, M. Bilinski1, D.Lorenzi1, H. Uriondo1, F, Noblía1, V. Longobucco1, E. Ventimiglia Lagar1, F. Nodar1
1Centro de Estudios en Genética y Reproducción (CEGYR), Buenos Aires, Argentina
Objective: The aim of this study was to investigate the effect of sperm DNA fragmentation on fertilization rate, embryo development (blastulation rate) and pregnancy outcomes for ICSI cycles in a cohort of couples that performed egg donation. Assess embryo quality (remaining embryos, not transferred or not frozen) for apoptotic markers.
Methods: For the present study, eighty-two (82) women who underwent to ART procedures (egg recipients) (2016) were selected. The recipient's average age was 41.8±5.1 y/o (36 - 49) and the oocyte donor's average age was 30.8±2.1 y/o (27-33). In our IVF center, egg donation cycles with frozen sperm samples are performed regularly, however 35 cycles were done using fresh sperm samples. The average male age was 40.1±5.2 y/o. Fertilization, blastulation and pregnancy rates were assessed. For the analysis two groups were considered, TUNEL <15% and ≥15%. In arrested embryos, ICC was performed to detect cleaved caspase 3, Survivin, TUNEL and DNA. The Student's t-test for between-group comparisons was used. Mann-Whitney U-test to assess homogeneity was used. Pearson correlation was also conducted. Differences were considered significant when P<0.05
Results: The present results demonstrated that exist a negative correlation (R=-0.5) between DNA fragmentation and the blastulation rate. These results evidenced that a high level of DNA fragmentation is associated with a low blastulation and pregnancy rate (per transfer), however the fertilization rate was not affected. We observed that samples with higher levels of DNA fragmentation induced higher levels of DNA fragmentation in blastomeres without activating the apoptosis pathway (9.1% vs. 15.9%) (P<0.05). Blastomeres from samples with high DNA fragmentation activated the apoptotic pathway in higher levels than TUNEL <15% (16.4% vs. 21.9%) (P<0.05).
Conclusion: Sperm DNA fragmentation has a negative correlation with blastulation and pregnancy rates even with good oocyte quality. High levels of DNA damage promotes embryo arrest inducing the apoptotic machinery.
C. Duarte1, V. Núñez1, Y. Wong1,2, C. Vivar1, E. Benites1, U. Rodriguez1, C. Vergara1, J. Ponce1
1NIU VIDA Specialized Center for Assisted Reproduction, Lima, Peru
2Laboratory of Animal Biotechnology, Ricardo Palma University, Lima, Peru
Soledad Sepúlveda Award
Best Oral Presentation.
13° REDLARA General Congress - Buenos Aires - Argentina 2017
Objective: In assisted reproduction procedures we need to develop and enhance new protocols to optimize the sperm selection in assisted reproduction procedures. The aim of this work is to evaluate the ability of the Z potential technique to select sperms with intact DNA in non-normospermic patients and evaluate the impact of this selection on embryonic development.
Methods: We analyzed a total of 174 human seminal samples with at least one altered parameter. We measured basal, post density gradients, and post density gradients + Z potential DNA fragmentation index. To evaluate the impact of this technique on embryo development, 54 cases were selected. The embryo development parameters evaluated were fertilization rate, cleavage rate, top quality embryos at the third day and blastocysts rate.
Results: We found significant differences in the studied groups when we compare the sperm fragmentation index by adding the Z potential technique to density gradient selection vs. density gradients solely. Furthermore, there is no significant difference in the embryo development parameters between low sperm fragmentation index group vs. moderate and high sperm fragmentation index groups when selecting sperms with this new technique.
Conclusions: Z potential technique is a very useful tool for sperm selection; it significantly reduces the DNA fragmentation index and improves the parameters of embryo development. This technique could be considered routine for its simplicity and low cost.
A. Delgado-Elías1,2, G. Llerena1, R. López3, J. Portella1, N. Inoue1, L. Noriega-Hoces1,2, L. Guzman1,3
1PRANOR Laboratorio. Grupo de Reproducción Asistida. San Isidro. Lima. Peru
2Clinica Concebir. San Isidro. Lima. Peru
3Reprogenetics Latinoamérica, Lima-Peru
Introduction: PGD for HLA typing is a procedure that can be performed when an affected child requires a transplant to treat a non-hereditary disorder related to the hematopoietic and/or immune system. Hematopoietic stem cell transplantation from an identical HLA-donor provides the best treatment option.
Case report: In Clínica Concebir San Isidro - Lima-Perú, three conventional ovarian stimulation procedures for IVF were performed in a couple with a 10-years-old child; which has a diagnosis of acute lymphatic T-cell leukemia of high risk. Trofoectoderm biopsy and aCGH examination were performed on 15 blastocysts, 3 on the first IVF procedure, 4 on the second cycle and 8 on the third. Three euploid blastocysts, HLA-compatible with the affected child genome were identified. A euploid blastocyst, HLA-compatible with the affected child was warmed and transferred; resulting in a live birth HLA match.
Comments: In conclusion, the combination of aCGH for aneuploidy screening and Karyomapping procedure could be performed in a single biopsy procedure.
M.T. Urbina1, I. Benjamin1, R. Medina1, J.Jiménez1, L. Trías1, J. Lerner1
1Unifertes Fertility Unit, Caracas, Venezuela
Objective: To discuss the implications of Expanded genetic carrier screening for preconception purposes based on our own practice.
Methods: One hundred and forty-three potential gamete donors aged 20-32 years old (x- = 24, 127 females and 16 males), signed informed consent and were selected according to REDLARA guidelines. Blood or saliva sample was examined by one of these genetic carrier screening: Genzyme screening for Cystic Fibrosis (CF), Fragile X and Spinal Muscular Atrophy (SMA); Counsyl Universal panel or Recombine Carrier Map.
Results: Genotyping results for all donors were analyzed; 41% (58/143) of donors were identified as carriers for at least one condition. We found a carrier frequency of (1/24) for CF, (1/72) for SMA and 0/120 for Fragile X syndrome. Among the high-impact most prevalent conditions in our study (Carrier Map group) were: 21-Hydroxilase-Deficient Congenital Nonclassical Adrenal Hyperplasia (1/8), Factor V deficiency (1/12), Hemochromatosis: Type 1: HFE Related (1/12), Short Chain Acyl-CoA (1/14) and MTHFR deficiency (1/3) (39%).
Conclusion: The rate of gamete donors identified as carriers for at least one condition was 41%, which supports the offering of Expanded carrier screening to our population. Studies in Latin American populations could help to customize screening panels.
The ART patient population has a unique opportunity to be offered Expanded carrier screening and appropriate counseling, to make its best-informed decisions.
Patient, health providers, psychologists and legal professional's education based on current scientific evidence; appropriate counseling about benefits, limitations, residual risk; generic informed consent and universal access are aspects to be considered.