Serum miR-181d-5p levels in response to controlled ovarian stimulation: predictive value and biological function.

The response to controlled ovarian stimulation (COS) for in vitro fertilization (IVF) varies dramatically from one patient to another, affecting success rates. A previous large-scale study identified increased serum miR-181d-5p levels in patients with high response to COS prior to stimulation. We aim to evaluate whether the expression of miR-181d-5p differs according to the ovarian response to COS in women undergoing intracytoplasmic sperm injection (ICSI) cycles. Samples collected prior to COS for ICSI were split into three groups depending on the ovarian response to COS: poor response (PR), <4 oocytes retrieved (n=25); normal response (NR), ≥8 and ≤12 oocytes retrieved (n=21); and high response (HR), >25 oocytes retrieved (n=20). miR-181d-5p serum levels were compared among experimental groups. miR-181d-5p levels were increased in the HR group when compared to the PR (p=0.0001) and NR groups (p=0.0079). miR-181d-5p levels correlated with the number of aspirated follicles (p<0.0001), retrieved oocytes (p<0.0001), and mature oocytes (p=0.0002). Increased miR-181d-5p levels independently predict a high response (p=0.006), with Positive and Negative Predictive Values of 66.7% and 69.4%, respectively. miR-181d-5p was also detected in the ovarian tissue in a mouse model. Moreover, computational analysis of miR-181d-5p predicted targets and promoter region suggested that this miRNA might be involved in the regulation of key signaling pathways and biological processes for female reproductive biology. In conclusion, miR-181d-5p is a promising circulating predictor of high stimulation and potential mediator of the hypothalamus-pituitary-gonad axis, providing opportunities for the individualization of COS protocols.

Serum metabolites as predictive molecular markers of ovarian response to controlled stimulation: a pilot study.

OBJECTIVE: This study aimed to look into the use of serum metabolites as potential biomarkers of response to controlled ovarian stimulation (COS) in patients undergoing intracytoplasmic sperm injection (ICSI) cycles. METHODS: This case-control study analyzed serum samples from 30 patients aged <36 years undergoing COS for ICSI in a university-affiliated assisted reproduction center from January 2017 to August 2017. The samples were split into three groups based on response to COS as follows: poor responders: <4 retrieved oocytes (PR group, n=10); normal responders: ≥ 8 and ≤ 12 retrieved oocytes (NR group, n=10); and hyper-responders: >25 retrieved oocytes (HR, n=10). The metabolic profiles of the serum samples were compared between the groups through Principal Component Analysis (PCA). Receiver Operating Characteristic (ROC) curves were built to assess the power of the model at predicting response to COS. RESULTS: PCA clearly distinguished between PR, NR and HR, and 10 ions were chosen as potential biomarkers of response to COS. These ions were more specific for PR than for NR. The ROC curve considering PR and NR had an area under the curve of 99.6% (95% CI: 88.9 - 100%). CONCLUSION: The preliminary evidence discussed in this study suggests that serum metabolites may be used as predictive molecular markers of ovarian response to controlled stimulation. The integration of clinical and "omics" findings may allow the migration toward an era of personalized treatment in reproductive medicine.

Lipidomic profile as a noninvasive tool to predict endometrial receptivity.

For the present study we asked whether the endometrial fluid lipidomic may be a useful approach to predict endometrial receptivity in freeze-all cycles. For this case-control study, endometrial fluid samples were collected from 41 patients undergoing freeze-all cycles. Samples were split depending on the pregnancy outcome: positive group (n = 24) and negative group (n = 17). Data were acquired by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were applied. A list of potential biomarker ion ratios was obtained and the values were used to build a receiver operating characteristic (ROC) curve to predict pregnancy success. The lipid categories were attributed by LIPID MAPS database. Ion ratios were established according to their correlations and used for the analysis. The PCA showed a tendency of separation between the studied groups, whereas the PLS-DA was able to clearly distinguish them. Fifteen ratios (13 hyper-represented in the negative and two hyper-represented in the positive group) were selected according to their importance for model prediction. These ratios were used to build the ROC curve, which presented an area under curve of 84.0% (95%CI: 69.2-97.4%; p = 0.009). These findings suggest that lipidomic profiling of endometrial fluid may be a valuable tool for identifying the time interval comprising the window of implantation.

Role of religion, spirituality, and faith in assisted reproduction.

Purpose: The purpose of this study is to evaluate the impact of the patient's faith, religion, and spirituality on the outcomes of intracytoplasmic sperm injection (ICSI) cycles. Materials and methods: Eight hundred and seventy-seven patients received a questionnaire containing information on faith, religiosity, and spirituality and the results of the questionnaires were correlated with ICSI outcomes. Patients stated to be Catholic (n = 476), spiritists (n = 93), Evangelical (n = 118), and other religion (n = 32), and 78 did not identify with any religious group. Results: A significant increase in fertilization, high-quality embryos, and pregnancy rate was found among Spiritists and Evangelicals. Patients who included the infertility diagnosis and treatment in their prayers showed an increased pregnancy rate, and those who reported their faith to be affected by the infertility diagnosis presented a decreased high-quality embryos rate. The high-quality embryos rate was increased among patients who answered that their faith contributed to their decision to undergo infertility treatment. The cycle's cancelation was negatively correlated with the frequency of religious meetings, and the frequency of prayers was positively correlated with the response to ovarian stimulation. Finally, belief in treatment success positively influenced the embryo quality. Conclusion: The findings suggest that spirituality plays a role in adjusting the psychological aspects of an infertile patient.

Paternal lifestyle factors in relation to semen quality and in vitro reproductive outcomes.

This prospective-cohort study aimed at investigating the influence of paternal lifestyle factors on semen parameters and intracytoplasmic sperm injection (ICSI) outcomes. The influence of paternal lifestyle factors on seminal quality and ICSI outcomes was investigated in male patients undergoing conventional semen analysis. Cigarette smoking negatively influenced semen volume (B: -0.417, slope: 1.570, p = 0.047), sperm count/ml (B: -7.363, slope: 52.298, p = 0.014), total sperm count (B: -4.43, slope: 178.165, p = 0.023), total motile sperm count (B: -1.38, slope: 100.276, p = 0.045) and SDF (B: 0.014, slope: 9.767, p = 0.033). Alcohol consumption negatively influenced sperm count/ml (B: -12.527, slope: 42.255, p = 0.040) and sperm DNA fragmentation (B: 5.833, slope: 9.680, p = 0.002). There were no significant influences of other paternal lifestyle factors. Cigarette smoking negatively influenced the fertilisation rate (B: -1.349, slope: 21.950, p = 0.039) and the blastocyst formation rate (B: -14.244, slope: 28.851, p = 0.025). Alcohol consumption negatively influenced fertilisation rate (B: -3.617, slope: 20.138, p = 0.041) and blastocyst formation rate (B: -34.801, slope: 30.044, p = 0.042). Cigarette smoking and alcohol consumption appear to reduce semen quality, fertilisation and blastocyst formation rates; thus, it would be wise to recommend that male partners reconsider their lifestyle during in vitro reproduction treatment.

Is there an association between artificial sweetener consumption and assisted reproduction outcomes?

Previous studies have suggested an association between high intake of sweetened beverages and a number of adverse health outcomes. In this cross-sectional study, we investigated the association between daily consumption of sweetened soft drinks or coffee and the outcome of intracytoplasmic sperm injection (ICSI) treatment. Patients (n = 524) were interviewed by a nutritionist before ICSI treatment, using a food frequency questionnaire. Regression analysis showed that consumption of ≥3 servings of regular soft drinks or any amount of diet soft drinks was associated with oocyte dysmorphism, diminished embryo quality on days 2 and 3 of culture, and a mild effect on blastocyst formation, implantation and pregnancy rate. Consumption of artificially sweetened coffee was negatively associated with embryo quality on days 2 and 3. However, consumption of coffee or soft drinks was not associated with the odds of live birth. Even so, patients should be advised about the potential negative effects of sugar and artificial sweeteners before attempting infertility treatment. This study is limited by the use of a non-validated food frequency questionnaire, lack of information on quantity of sweeteners consumed, and lack of data on glucose levels in blood serum or follicular fluid. Further investigation is warranted.

FSH dose to stimulate different patient' ages: when less is more.

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 we found that the lower the age, the lower the FSH dose needed per oocyte retrieved. In the group of patients ≤35 y.o., we also found a positive effect of the FSH dose on oocyte yield. Despite that, for patients ≤38 y.o. there was a negative effect of the FSH dose on embryo quality and blastocyst formation rate, and an increase in the cycle's cancelation rate. In patients ≥39 y.o., there were no effects of the FSH doses on the analysed variables. CONCLUSIONS: Ovarian stimulation with high doses of FSH is not recommended in younger women (≤38 y.o.), once we found 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 a higher FSH dose is needed for oocyte retrieval in these patients.

Overcoming male factor infertility with intracytoplasmic sperm injection.

OBJECTIVE: To evaluate the effect of male factor infertility on intracytoplasmic sperm injection (ICSI) outcomes compared with a control group presenting isolated tubal factor. METHOD: This retrospective study included 743 couples undergoing ICSI as a result of isolated male factor and a control group consisting of 179 couples undergoing ICSI as a result of isolated tubal factor, performed in a private university- -affiliated in vitro fertilization center, between January/2010 and December/2016. Patients were divided into two groups according to maternal age: women ≤35 years old and >35 years old. The effects of infertility causes on laboratorial and clinical ICSI outcomes were evaluated using Student's t-test and (2 test. RESULTS: No differences in controlled ovarian stimulation outcomes were observed between male factor cycles and tubal factor cycles in the two age groups. Implantation (male factor 35.5% vs. tubal factor 32.0%, p=0.340), pregnancy (male factor 46.9% vs. tubal factor 40.9%, p=0.184) and miscarriage (male factor 10.3% vs. tubal factor 10.6%, p=0.572) rates were similar between the infertility groups, irrespective of female age. Considering maternal age, the cancelation rate was higher in older women (>35 years old) undergoing ICSI as a result of male factor infertility (17.4% vs. 8.9%, p=0.013). CONCLUSION: Our results showed that there is no difference in the outcomes of pregnancy between couples with male or tubal factor infertility, which indicates that ICSI surpasses the worse specific outcomes associated with male factor.

The predictive value of serum concentrations of anti-Müllerian hormone for oocyte quality, fertilization, and implantation.

OBJECTIVE: This study aimed to identify a possible correlation between serum levels of anti-Müllerian hormone (AMH) and oocyte quality, embryo developmental competence, and implantation potential. METHODS: 4488 oocytes obtained from 408 patients undergoing ICSI cycles were evaluated. Oocyte dimorphisms, embryo quality on days two and three, blastocyst formation competence, fertilization rates, implantation rates, and pregnancy rates were correlated with serum levels of AMH using Pearson's correlation coefficient and regression analysis. RESULTS: A positive correlation was observed between serum levels of AMH and number of retrieved oocytes (CC: 0.600, p<0.001), fertilization rate (CC:0.595, p=0.048), and number of obtained embryos (CC:0.495, p<0.001). AMH did not affect the quality of cleavage stage embryos or the chance of blastocyst formation. However, AMH levels affected oocyte quality (OR:0.75, CI 0.44-0.96, p<0.001), and implantation (CC:0,116, p=0.031) and pregnancy (OR:1.22, CI:1.03-1.53, p<0.001) rates. CONCLUSION: Serum levels of AMH are a useful predictor of ovarian response to COS, oocyte quality, and fertilization. However, AMH levels may also compromise clinical outcomes; lower AMH levels did not impair embryo development.

Overcoming male factor infertility with intracytoplasmic sperm injection / Superando o fator masculino de infertilidade com injeção intracitoplasmática de espermatozoides

Summary Objective: To evaluate the effect of male factor infertility on intracytoplasmic sperm injection (ICSI) outcomes compared with a control group presenting isolated tubal factor. Method: This retrospective study included 743 couples undergoing ICSI as a result of isolated male factor and a control group consisting of 179 couples undergoing ICSI as a result of isolated tubal factor, performed in a private university- -affiliated in vitro fertilization center, between January/2010 and December/2016. Patients were divided into two groups according to maternal age: women ≤35 years old and >35 years old. The effects of infertility causes on laboratorial and clinical ICSI outcomes were evaluated using Student's t-test and (2 test. Results: No differences in controlled ovarian stimulation outcomes were observed between male factor cycles and tubal factor cycles in the two age groups. Implantation (male factor 35.5% vs. tubal factor 32.0%, p=0.340), pregnancy (male factor 46.9% vs. tubal factor 40.9%, p=0.184) and miscarriage (male factor 10.3% vs. tubal factor 10.6%, p=0.572) rates were similar between the infertility groups, irrespective of female age. Considering maternal age, the cancelation rate was higher in older women (>35 years old) undergoing ICSI as a result of male factor infertility (17.4% vs. 8.9%, p=0.013). Conclusion: Our results showed that there is no difference in the outcomes of pregnancy between couples with male or tubal factor infertility, which indicates that ICSI surpasses the worse specific outcomes associated with male factor.

Resumo Objetivo: Avaliar o efeito do fator masculino de infertilidade em resultados de injeção intracitoplasmática de espermatozoides (ICSI) em comparação com um grupo controle que apresenta o fator tubário isolado. Método: Este estudo retrospectivo incluiu 743 casais submetidos a ICSI por fator masculino e 179 casais por fator tubário, realizada em um centro privado de fertilização in vitro associado à universidade, entre janeiro de 2010 e dezembro de 2016. Os pacientes foram divididos em dois grupos de acordo com a idade materna: mulheres ≤ 35 e > 35 anos de idade. Os efeitos das causas de infertilidade nos resultados laboratoriais e clínicos da ICSI foram avaliados pelos testes T de Student e Qui-quadrado. Resultados: Não foram observadas diferenças nos parâmetros de estimulação ovariana entre os ciclos com fatores masculinos e com fatores tubários. A taxa de implantação (fator masculino 35,5% vs. fator tubário 32,0%, p=0,340), de gravidez (fator masculino 46,9% vs. fator tubário 40,9%, p=0,184) e de aborto (fator masculino 10,3% vs. fator tubário 10,6%, p=0.572) foram semelhantes entre os grupos de infertilidade, independentemente da idade feminina. Considerando a idade materna, a taxa de cancelamento foi maior em mulheres > 35 anos cuja causa de infertilidade era o fator masculino (17,4% vs. 8,9%, p=0,013). Conclusão: Não há diferenças nos resultados de gravidez entre casais com infertilidade dos fatores masculino ou tubário isolados, o que indica que ICSI supera os piores resultados associados ao fator masculino.

miR-142-3p as a biomarker of blastocyst implantation failure - A pilot study.

OBJECTIVE: This study aims to find whether microRNAs (miRNAs) detected in the culture medium of embryos produced in vitro could be potential biomarkers of embryo implantation. METHODS: Culture media samples from 36 embryos, derived from patients undergoing intracytoplasmic sperm injection (ICSI) in a private university-affiliated IVF center, were collected between January/2015 and November/2015. Samples were collected on day three and embryo transfers were performed on day five and all embryos reached the blastocyst stage. Samples were split into groups according to the embryo implantation result: Positive-Implantation-Group (n=18) or Negative-Implantation-Group (n=18). For the first analysis, samples were pooled in three sets for each group (6-7 spent media per pool). MicroRNAs were extracted from spent media and cDNA was synthesized. C. elegans miR-39 was used as RNA spike-in to normalize the gene expression analysis. The expression of microRNAs into the spent media from the Positive-Implantation-Group was compared with those from the Negative-Implantation-Group. A set of seven miRNAs (miR-21, miR-142-3p, miR-19b, miR-92a, miR-20b, miR-125a and miR148a) selected according with the literature, was tested. To check whether miRNAs could be detected in individual samples of culture media, in a second analysis, ten more samples were tested for miR-21 and miR-142-3p. RESULTS: From the sevens tested miRNAs, a significant increased expression of miR-142-3p could be noted in the Negative-Implantation-Group (P<0.001). For other three miRNAs (miR-21, miR-19b and miR-92a) a difference in expression was observed, however it did not reach a statistical significance. In addition, when ten non-redundant samples were tested to check if miRNAs could be detected in individual samples of culture media, the highly specific amplification of mature miRNAs, including miR-142-3p, could be noted. CONCLUSION: Our findings suggest that miR-142-3p, previously described as a tumor suppressor and cell cycle inhibitor, may be a potential biomarker of blastocyst implantation failure. The identification of miRNAs on individual culture medium samples offers unique opportunities for non-invasive early diagnosis of blastocyst implantation.

[Limitations and controversies in determining the predictive value of oocyte and embryo morphology criteria]. / Limitações e controvérsias na determinação do valor preditivo de critérios de morfologia oocitária e embrionária.

In order to increase the success rate of in vitro fertilization cycles, several studies have focused on the identification of the embryo with higher implantation potential. Despite recent advances in the reproductive medicine, based on the OMICs technology, routinely applicable methodologies are still needed. Thus, in most fertilization centers embryo selection for transfer is still based on morphological parameters evaluated under light microscopy. Several morphological parameters may be evaluated, ranging from the pronuclear to blastocyst stage. In general, despite the day of transfer, some criteria are suggested to present a predictive value for embryo viability when analyzed independently or combined. However, the subjectivity of morphological evaluation, as well as the wide diversity of embryo classification systems used by different fertilization centers shows contrasting results, making the implementation of a consensus regarding different morphological criteria and their predictive value a difficult task. The optimization of embryo selection represents a large potential to increase treatment success rates, allowing the transfer of a reduced number of embryos and minimizing the risks of multiple pregnancy.

Limitações e controvérsias na determinação do valor preditivo de critérios de morfologia oocitária e embrionária / Limitations and controversies in determining the predictive value of oocyte and embryo morphology criteria

Com o objetivo de aumentar as taxas de sucesso das pacientes que são submetidas a técnicas de reprodução humana assistida (RHA), numerosos estudos apresentam como foco a identificação do embrião com maior potencial de implantação. Apesar dos avanços tecnológicos significativos da Medicina Reprodutiva baseados no advento da era da genômica, proteômica e metabolômica (OMICS), técnicas rotineiramente aplicáveis ainda não estão disponíveis. Dessa forma, laboratórios de fertilização in vitro(FIV) de todo o mundo selecionam para transferência embriões humanos cultivados in vitrobaseados em parâmetros morfológicos avaliados em microscopia de luz. Diversos parâmetros morfológicos podem ser avaliados desde o estágio pronuclear até o estágio de blastocisto para embriões humanos cultivados in vitro. De modo geral, independentemente do dia da transferência, tais critérios parecem apresentar valor preditivo de viabilidade embrionária quando avaliados individualmente ou coletivamente. No entanto, a subjetividade da avaliação morfológica, bem como a ampla diversidade de sistemas de classificação embrionária aplicados por diferentes clínicas, implica em resultados contraditórios, tornando extremamente difícil a implementação de um consenso do valor preditivo dos diferentes parâmetros morfológicos avaliados. A otimização da seleção embrionária representa um grande potencial de aumento das taxas de sucesso do tratamento, além de possibilitar a realização da transferência de um número reduzido de embriões, minimizando os riscos derivados de estações múltiplas.

In order to increase the success rate of in vitrofertilization cycles, several studies have focused on the identification of the embryo with higher implantation potential. Despite recent advances in the reproductive medicine, based on the OMICs technology, routinely applicable methodologies are still needed. Thus, in most fertilization centers embryo selection for transfer is still based on morphological parameters evaluated under light microscopy. Several morphological parameters may be evaluated, ranging from the pronuclear to blastocyst stage. In general, despite the day of transfer, some criteria are suggested to present a predictive value for embryo viability when analyzed independently or combined. However, the subjectivity of morphological evaluation, as well as the wide diversity of embryo classification systems used by different fertilization centers shows contrasting results, making the implementation of a consensus regarding different morphological criteria and their predictive value a difficult task. The optimization of embryo selection represents a large potential to increase treatment success rates, allowing the transfer of a reduced number of embryos and inimizing the risks of multiple pregnancy.

Decline in semen quality among infertile men in Brazil during the past 10 years.

PURPOSE: To investigate whether the semen quality of men undergoing conventional semen analysis is deteriorating over time. MATERIALS AND METHODS: We analyzed and compared the sperm count, motility and morphology of 2300 semen samples provided by males undergoing conventional seminal analysis, from years 2000 to 2002 and 2010 to 2012. The incidences of severe oligozoospermia and azoospermia over time were also compared. RESULTS: A total of 764 sperm samples were analyzed in 2000-2002 and 1536 in 2010-2012. Over time, the mean sperm concentration/ml decreased significantly from 61.7 million in 2000-2002 to 26.7 million in 2010-2012 (R2 = 11.4%, p < 0.001), the total sperm concentration decreased significantly from 183.0 million to 82.8 million (R2 = 11.3%, p < 0.001), and the percentage of normal forms decreased significantly from 4.6% to 2.7% (R2 = 9.8%, p < 0.001). The incidence of severe oligozoospermia significantly increased from 15.7% to 30.3% (OR: 1.09, p < 0.001) and the incidence of azoospermia increased from 4.9% to 8.5% (OR: 1.06, p = 0.001). CONCLUSIONS: This study demonstrated a significant time-related decline in semen quality of infertile patients. This finding might have implications on fertility and emphasizes the need for further studies addressing subject's life-style in order to find and reduce the causative agents. Future prospective and multicenter studies including representative samples of the general population are needed to confirm whether semen quality is really declining.

Blastocyst Morphology Holds Clues Concerning The Chromosomal Status of The Embryo.

BACKGROUND: Embryo morphology has been proposed as an alternative marker of chro- mosomal status. The objective of this retrospective cohort study was to investigate the association between the chromosomal status on day 3 of embryo development and blas- tocyst morphology. MATERIALS AND METHODS: A total of 596 embryos obtained from 106 cycles of intra- cytoplasmic sperm injection (ICSI) followed by preimplantation genetic aneuploidy screening (PGS) were included in this retrospective study. We evaluated the relation- ship between blastocyst morphological features and embryonic chromosomal altera- tion. RESULTS: Of the 564 embryos with fluorescent in situ hybridization (FISH) results, 200 reached the blastocyst stage on day 5 of development. There was a significantly high- er proportion of euploid embryos in those that achieved the blastocyst stage (59.0%) compared to embryos that did not develop to blastocysts (41.2%) on day 5 (P<0.001). Regarding blastocyst morphology, we observed that all embryos that had an abnormal inner cell mass (ICM) were aneuploid. Embryos with morphologically normal ICM had a significantly higher euploidy rate (62.1%, P<0.001). As regards to the trophectoderm (TE) morphology, an increased rate of euploidy was observed in embryos that had nor- mal TE (65.8%) compared to embryos with abnormal TE (37.5%, P<0.001). Finally, we observed a two-fold increase in the euploidy rate in high-quality blastocysts with both high-quality ICM and TE (70.4%) compared to that found in low-quality blastocysts (31.0%, P<0.001). CONCLUSION: Chromosomal abnormalities do not impair embryo development as ane- uploidy is frequently observed in embryos that reach the blastocyst stage. A high-quality blastocyst does not represent euploidy of chromosomes 13, 14, 15, 16, 18, 21, 22, X and Y. However, aneuploidy is associated with abnormalities in the ICM morphology. Further studies are necessary to confirm whether or not the transfer of blastocysts with low-quality ICM should be avoided.

Decline in semen quality among infertile men in Brazil during the past 10 years

ABSTRACTPurpose:To investigate whether the semen quality of men undergoing conventional semen analysis is deteriorating over time.Materials and Methods:We analyzed and compared the sperm count, motility and morphology of 2300 semen samples provided by males undergoing conventional seminal analysis, from years 2000 to 2002 and 2010 to 2012. The incidences of severe oligozoospermia and azoospermia over time were also compared.Results:A total of 764 sperm samples were analyzed in 2000-2002 and 1536 in 20102012. Over time, the mean sperm concentration/ml decreased significantly from 61.7 million in 2000-2002 to 26.7 million in 2010-2012 (R2=11.4%, p<0.001), the total sperm concentration decreased significantly from 183.0 million to 82.8 million (R2=11.3%, p<0.001), and the percentage of normal forms decreased significantly from 4.6% to 2.7% (R2=9.8%, p<0.001). The incidence of severe oligozoospermia significantly increased from 15.7% to 30.3% (OR: 1.09, p<0.001) and the incidence of azoospermia increased from 4.9% to 8.5% (OR: 1.06, p=0.001).Conclusions:This study demonstrated a significant time-related decline in semen quality of infertile patients. This finding might have implications on fertility and emphasizes the need for further studies addressing subject's life-style in order to find and reduce the causative agents. Future prospective and multicenter studies including representative samples of the general population are needed to confirm whether semen quality is really declining.

The impact of the embryo quality on the risk of multiple pregnancies.

The aim of the present study was to determine the chance of pregnancy and the risk of multiple pregnancies taking into account the number and quality of transferred embryos in patients >36 years old or ≤36 years old. For this study, 1497 patients undergoing intra-cytoplasmic sperm injection (ICSI) cycles in a private assisted reproduction centre were split into groups according to the number and quality of the transferred embryos on the third or fifth day of development. The pregnancy rate and multiple pregnancy rate were compared between the embryo quality groups in patients <36 years old or ≥36 years old. In patients <36 years old, for the day 3 embryo transfer, no significant difference was noted in the pregnancy rate when the groups were compared. However the multiple pregnancy rate was increased by the transfer of an extra low-quality embryo (17.1 versus 28.2%, P = 0.020). For day 5 embryo transfer, the transfer of an extra blastocyst significantly increased the pregnancy rate (36.0 versus 42.4%, P < 0.001) and the multiple pregnancy rate (4.4 versus 16.9%, P < 0.001). In older patients, no significant difference was noted in the pregnancy rate when the groups were compared. However, when an extra low-quality embryo was transferred, a significantly increased rate of multiple pregnancies was observed for day 3 (18.2 versus 26.4%, P = 0.049) and day 5 embryo transfers (5.2 versus 16.1%, P < 0.001). In conclusion, the transfer of an extra low-quality embryo may increase the risk of a multiple pregnancy. In younger patients, the transfer of an extra low-quality blastocyst may also increase the chance of pregnancy.

The efficiency of a donor-recipient program using infertile donors' egg cryo-banking: a Brazilian reality.

PURPOSE: To determine whether Brazilian egg donation treatment outcomes with oocytes donated from infertile couples are equivalent to those obtained worldwide with oocytes donated from fertile egg-donors. METHODS: In this descriptive study, egg-donation cycles from 259 women, performed from January 2009 to July 2013, were evaluated. Oocytes were obtained from patients undergoing ICSI who decided to donate their surplus oocytes. We described the survival, fertilization, blastocyst, implantation and pregnancy rates obtained in our infertile donor-recipient program. In addition, we described the results obtained in previous published studies. RESULTS: In our egg-donation program we obtained a fertilization rate of 72.9 %, a blastocyst formation rate of 53.2 %, an implantation rate of 31.1 % and the estimated clinical pregnancy rate per warmed oocyte was 5.4 %. The analyzed studies, performed between 2008 and 2013, included varying numbers of egg-donors (range: 20-600), warmed oocytes (range: 123-3826) and survival rates (range: 85.6-92.5 %). Fertilization rates ranged from 74.2 to 87.0 %, blastocyst formation rate ranged from 41.3 % to 68.0 %, implantation rates ranged from 24.7 % to 55.3 % and the clinical pregnancy rate per warmed oocyte ranged from 3.9 % to 9.8 %. CONCLUSIONS: New and reassuring information derived from our egg-donation program demonstrates outcomes similar to those reported for other egg donation programs.

Influence of oocyte dysmorphisms on blastocyst formation and quality.

OBJECTIVE: To identify the association between oocyte dysmorphisms and blastocyst developmental competence. DESIGN: Case-control study. SETTING: Private assisted reproduction center. PATIENT(S): This study included 5,516 embryos, which were obtained from 605 patients undergoing intracytoplasmic sperm injection cycles in which ET was performed on day 5 of embryo development. INTERVENTION(S): Intracytoplasmic sperm injection and ET. MAIN OUTCOME MEASURE(S): The morphologic characteristics of the blastocysts, namely the degree of blastocoel expansion and hatching status, the size and compactness of the inner cell mass (ICM), and the cohesiveness and number of trophectoderm cells, were correlated with oocyte dysmorphisms. RESULT(S): Of the 5,516 embryos evaluated, 2,920 (52.9%) reached the blastocyst stage. The blastocysts' degree of expansion and hatching status were decreased by the presence of aggregates of smooth endoplasmic reticulum clusters, large perivitelline space, and shape abnormalities. The presence of a large perivitelline space also decreased the ICM and trophectoderm cell quality. Aggregates of smooth endoplasmic reticulum clusters also decreased the ICM quality, pregnancy rate, and the miscarriage rate. CONCLUSION(S): The results presented here suggest that the individual identification of oocyte dymorphisms may be a prognostic tool for blastocyst development and quality. However, the results do not diminish the importance of cleavage-stage embryo morphology assessment to predict blastocyst development competence.

Intracytoplasmic morphologically selected sperm injection outcomes: the role of sperm preparation techniques.

PURPOSE: To compare the results of intracytoplasmic morphologically selected sperm injection (IMSI) between cycles in which the swim-up (SUP) or the density gradient centrifugation (DGC) techniques were used for sperm preparation. METHODS: We evaluated 70 IMSI cycles performed in women with age ≤ 37 years, undergoing IMSI as result of male factor. The couples were divided into two groups: DGC group (n = 26) and SUP group (n = 44). The groups were compared with regard to IMSI outcomes. RESULTS: There were no significant differences between SUP and DGC groups regarding the number of follicles, oocytes, mature oocytes, oocyte yield and mature oocyte rate. Fertilization rate and high-quality embryos rate on day 5 of development were similar between SUP and DGC groups. Implantation, pregnancy and miscarriage rates were not statistically different between SUP and DGC groups (28.8 vs 33.3 %, 46.2 vs 57.1 % and 8.3 vs 4.2 %, respectively). CONCLUSIONS: Both the SUP and the DGC techniques recover improved sperm fractions and result in similar IMSI outcomes. Further randomized trials analyzing both the quality of sperm through MSOME and the IMSI outcomes are needed to elucidate the role of sperm preparation techniques and morphology on IMSI outcomes.