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.

Previous infection with SARS-CoV-2 impacts embryo morphokinetics but not clinical outcomes in a time-lapse imaging system.

The goal for the present study was to investigate whether previous infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may compromise embryo morphokinetics and implantation. For that, a historical cohort study was performed in a private university-affiliated in vitro fertilization center. The study included 1628 embryos from 88 patients undergoing intracytoplasmic sperm injection (ICSI) cycles. Patients were age-matched in a 1:3 ratio to either a coronavirus disease (COVID) group, including patients with a positive SARS-CoV-2 immunoglobulin test (n = 22 patients, 386 embryos), or a control group, including patients with a negative SARS-CoV-2 immunoglobulin test (n = 66, 1242 embryos). The effect of previous infection with SARS-CoV-2 on morphokinetic events and ICSI outcomes was evaluated. Embryos derived from patients in the COVID group presented longer time to pronuclei appearance and fading, time to form two, three, four and five cells, and time to blastulation. The durations of the third cell cycle and to time to complete synchronous divisions were also significantly increased in the COVID group compared with the control group, whereas known implantation diagnosis score Day 5 ranked significantly lower in the COVID group. No differences were observed between the COVID and control groups on clinical outcomes. In conclusion, patients planning parenthood, who have recovered from COVID-19 infection, must be aware of a possible effect of the infection on embryo development potential.

Metabolomic profile as a noninvasive adjunct tool for the diagnosis of Grades III and IV endometriosis-related infertility.

The aim of the present case-control study was to develop a noninvasive adjuvant tool for the diagnosis of endometriosis. Serum samples from 100 patients undergoing intracytoplasmic sperm injection were split into two groups according to the cause of infertility: an endometriosis group (n = 50), consisting of samples derived from patients with Grade III and IV endometriosis, and a control group (n = 50), comprising samples derived from patients with isolated male factor infertility. The metabolomic profile of each sample was obtained, through mass spectrometry. Partial least squares discriminant analysis was able to clearly classify the endometriosis and control groups. Ten potential biomarkers were selected based on their importance for model prediction. These ions were used to build the receiver-operating characteristic curve, which presented an area under the curve of 0.904 (95% confidence interval: 0.796-0.985). To validate the model, 30 other samples from infertile women without any evidence of endometriosis were tested. Considering these ions as possible biomarkers, the model was able to correctly classify 84% of the patients. Finally, a similar prediction potential was observed in the model validated set, when samples from the disease-free group were tested. Serum metabolomics may be useful as a noninvasive adjunct tool for the selection of patients who must undergo laparoscopy for definitive endometriosis diagnosis.

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.

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.

Non-invasive prediction of blastocyst implantation, ongoing pregnancy and live birth, by mass spectrometry lipid fingerprinting.

OBJECTIVE: To identify lipid markers of blastocyst implantation and ongoing pregnancy by day three culture medium mass spectrometry (MS) fingerprinting. METHODS: For this study, 33 culture media samples were harvested on day three, from 22 patients undergoing day five embryo transfers. All embryos achieved the blastocyst stage and were split into groups based on their implantation (Negative Implantation, n= 14 and Positive Implantation, n= 19). The positive implantation cycles resulted in successful ongoing pregnancies. The lipid extraction was performed by the Bligh-Dyer protocol and mass spectra were obtained with a direct infusion into a Q-Tof mass spectrometer. The data obtained was analyzed by Principal Component Analysis (PCA) and Partial Least Square Discrimination Analysis (PLS-DA). The statistical analysis was performed using the Metabo-Analyst 2.0. RESULTS: The variable importance in the projection (VIP) plot of the PLS-DA provided a list of four ions, in the positive mode, with an area under the curve (AUC) of 73.5%; and eight ions, in the negative mode, with and AUC of 72.0%. For both positive and negative modes, possible biomarkers for the negative implantation were identified by the lipidmaps: phosphoethanolamine, dicarboxylic acids, glycerophosphoglycerol, glycerophosphocholine, glicerophosphoinositol, phosphoethanolamine and unsaturated fat acids. The other ions were not identified. These lipids are involved in the GPI anchor biosynthesis and synthesis of lycerophospholipids and phosphate inositol. CONCLUSION: MS fingerprinting is useful to identify blastocysts that fail to implant, and therefore this technique could be incorporated into the laboratory routine, adjunct to morphology evaluation to identify embryos that should not be transferred.

Endometriosis Affects Oocyte Morphology in Intracytoplasmic Sperm Injection Cycles?

OBJECTIVE: To identify associations between presence of endometriosis and oocyte defects, embryo developmental potential, and cycle outcomes. METHODS: This study looked into the impact of endometriosis on oocyte and embryo quality, and blastocyst formation probability. Endometriosis was also correlated with cycle characteristics. In order to avoid age-related bias, in the first analysis only patients aged 36 years or younger were included, and the cycles were split into endometriosis infertility cycles (n=431; 3172 oocytes) and other cycles (n=2510; 24480 oocytes). RESULTS: The number of retrieved oocytes (10.6±21.2 vs. 14.6±21.1, P<0.001), oocyte yield (68.1±20.0% vs. 70.6±19.6%, P=0.015), and embryos obtained (6.1±4.43 vs. 7.8±5.12, P<0.001) were lower among patients with endometriosis. Implantation rates (28.1%±38.9% vs. 33.9±42.7, P<0.001) were lower among patients with endometriosis, but fertilization, pregnancy, miscarriage and cycle cancelation rates were not different. There was a significant increase in the incidence of extra-cytoplasmic, but not intra-cytoplasmic, oocyte defects among patients with endometriosis. The quality of embryos (45.3% vs. 47.3%, P=0.037) collected from patients with endometriosis was lower, but blastocyst formation rates were unaltered. CONCLUSION: A possible explanation for the lower implantation rates seen in patients with endometriosis is the poorer quality of the oocytes and embryos observed in this group of patients.

Does the Number of ICSI Cycles Performed per day or the Number of Oocytes Injected per day has an Impact on the Cycles' Outcome?

OBJECTIVE: To evaluate if ICSI outcome are influenced by the number of cycles and oocyte injected per day. METHODS: Data of ICSI cycles performed in an unselected infertile population were included. The influences of the number of cycles and number of injected oocytes on the ICSI outcomes (fertilization rate, high quality embryos rates on day 3 of development, blastocyst formation rate, the proportion of cycles with embryo transfer, pregnancy rate and implantation rate) were investigated. RESULTS: Regression analyses results showed no significant influence of the number of cycles and oocytes injected per day on the ICSI outcomes. CONCLUSION: The outcomes of ICSI are not influenced by the number of cycles or the number of oocytes injected per day. We suggest that an appropriate number of embryologists per cases per year guarantee successful outcomes independently of the daily workload.

Intracytoplasmic morphologically selected sperm injection results in improved clinical outcomes in couples with previous ICSI failures or male factor infertility: a meta-analysis.

The objective of this study was to perform the first meta-analysis to compare conventional intracytoplasmic sperm injection (ICSI) outcomes and intracytoplasmic morphologically selected sperm injection (IMSI) outcomes in couples with previous ICSI failures (IF) or male factor infertility (MF). A systematic review was performed by searching Medline database to identify articles reporting on the comparison between ICSI and IMSI outcomes in couples with IF or MF. The main outcome measures were the implantation, pregnancy and miscarriage rates. Thirteen studies fulfilled our predetermined criteria. The overall results of meta-analysis for implantation (OR: 2.88; CI: 2.13-3.89), pregnancy (OR: 2.07; CI: 1.22-3.50) and miscarriage rates (OR: 0.31; CI: 0.14-0.67) were in favor of IMSI in couples with IF. Additionally, the overall result of meta-analysis for implantation (OR: 1.56; CI: 1.11-2.18) and pregnancy rate (OR: 1.61; CI: 1.17-2.23) were in favor of IMSI in couples with MF. IMSI increases the odds of implantation by 50% and pregnancy by 60% in couples with MF. In light of improved clinical outcomes, we recommend promoting the IMSI method in couples with MF. Moreover, IMSI results in a 3-fold increase in implantation rate, a 2-fold increase in pregnancy rate and a 70% decrease in miscarriage rate as compared to ICSI in couples with IF, however, as no randomized evidence exists, randomized studies are needed to confirm the IMSI benefits in couples with IF.

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.

The importance of the cleavage stage morphology evaluation for blastocyst transfer in patients with good prognosis.

PURPOSE: To evaluate: (i) the influence of morphology at cleavage stage on blastocyst formation and implantation, and (ii) whether the transfer of low-quality embryos on day-three would be a better approach than the transfer at blastocyst stage. METHODS: This study included 8,444 embryos obtained from 1,125 patients undergoing ICSI cycles between January/2011 and September/2013. The influence of the quality of the embryo on days-two and -three on blastocyst formation and implantation success was evaluated. Moreover, the implantation potential of low-quality embryos, at cleavage stage, transferred on day-three was compared with the implantation potential of low-quality embryos, at cleavage stage, transferred on day-five. RESULTS: Low-quality embryos on day-two had an approximate 20 % decreased chance of achieving the blastocyst stage, and blastocysts derived from low-quality embryos on day-two had a nearly 40 % decrease in the implantation chance. Low-quality embryos on day-three had a 30 % decreased chance of achieving the blastocyst stage, and blastocysts derived from low-quality embryos on day-three had an almost 40 % decreased implantation chance. The implantation rate didn't differ when low-quality embryos on the cleavage stage were transferred on day-three or left in culture and transferred on day-five. CONCLUSIONS: The transfer of low-quality embryos on day-three is a better approach than transfer at the blastocyst stage. In addition, the embryo morphology evaluation at the cleavage stage is still needed for the selection of the embryo with the best implantation potential in extended embryo culture programmes.

Pituitary suppression with a GnRHa short protocol in an alternate day schedule associated with rhCG microdoses.

OBJECTIVE: To evaluate a comfortable short protocol with GnRH agonist (GnRHa) in alternate days, with a step down method of gonadotropins administration associated with hCG microdose for young patients undergoing intracytoplasmic sperm injection. METHODS: This study evaluated 89 ICSI cycles performed in female patients aged <36 years. Patients were submitted to a short protocol with GnRHa schedule in the study group (n= 25) and to a long pituitary suppression protocol in the control group (n=64). RESULTS: The total dose of rFSH administered as well as estradiol levels on the day of hCG trigger were significantly lower in the short protocol group. There were no significant differences between the groups regarding the fertilization, high-quality embryos, implantation, pregnancy and miscarriage rates. However, mean ovarian stimulation cost (GnRHa short group: $2,397 ± $870.3 and control group: $3,197 ± $658.9, P <0.001) and mean ovarian stimulation cost per pregnancy (GnRHa short group: ($4,993 ± $1,813 and control group: $9,743 ± $2,008, P <0.001) were significantly lower in the GnRHa short group as compared to the control group. CONCLUSION: In patients with normal ovarian response, pituitary suppression with a GnRHa short protocol in alternate days is less costly, requires lower gonadotropins doses and results in similar implantation and pregnancy rates as compared to a GnRHa long protocol.

The knowledge of the increased risk of complications in multiple pregnancies does not affect the desire to transfer more than one embryo in in vitro fertilisation treatment.

OBJETIVE: One of the main complications in in vitro fertilisation (IVF) is multiple pregnancies. This study was designed to investigate how many embryos subjects participating in an online survey would want to transfer in their IVF cycles. METHODS: This study was conducted in a Brazilian private assisted fertilisation centre. Individuals who accessed the centre's website were asked to participate in the survey. The survey was based on important information concerning multiple gestations, followed by a single multiple choice question, as follows: 'Knowing that the transfer of one embryo reduces the chance of pregnancy, and that the transfer of more than one embryo could result in multiple pregnancies, which comes with risks to the mother and the babies, answer: how many embryos would you transfer in your IVF cycle?'. There were three available answers: one, two or three embryos. RESULTS: A total of 1,049 subjects participated in the survey: 109 males and 940 females. The majority of the participants answered that they would like to have two embryos transferred (53.7%); followed by three embryos (35.0%), and one embryo (11.3%). CONCLUSION: Men and women tend to underestimate the risks of complications associated with multiple embryo transfers and multiple gestations. It is the physician's responsibility to consider single embryo transfer (SET) as the method of choice and perform double or triple embryo transfers only in special circumstances.

Intrafollicular soluble receptor for advanced glycation end products (sRAGE) and embryo quality in assisted reproduction.

The developmental potential of human embryos has important implications in assisted reproduction and depends, among other factors, on oocyte competency. The receptor for advanced glycation end products (RAGE) is a member of the superfamily of immunoglobulin cell-surface molecules that are constitutively expressed during embryonic development. RAGE is down-regulated in homeostasis in adult life. This study measured the concentration of soluble RAGE (sRAGE) in follicular fluid obtained from the leading follicle after ovarian stimulation of 54 women undergoing intracytoplasmic sperm injection. Corresponding embryos and sRAGE concentrations in follicular fluid were evaluated and correlations were investigated by multi-adjusted regression analysis. High intrafollicular sRAGE concentrations predicted poor-quality embryos (n=45, OR=0.986; P=0.026), adjusted for patient age, body mass index and oocyte quality, showing an inverse association between intrafollicular sRAGE concentrations and embryo development. The developmental potential of human embryos has important implications in assisted reproduction, and it depends, among other factors, on oocyte competency. The receptor for advanced glycation end products (RAGE) is a molecule constitutively expressed during embryonic development, but it is down-regulated in adult life. RAGE is frequently associated with pro-inflammatory responses, and it is implicated as an underlying condition in immune disorders, cardiovascular diseases, diabetes, Alzheimer's disease and cancer. In addition to activating the pro-inflammatory responses, RAGE down-regulates cellular defence mechanisms. The present study measured the concentrations of soluble RAGE (sRAGE) in follicular fluid samples obtained from leading follicles of women undergoing intracytoplasmic sperm injection (ICSI). This prospective cohort study included 54 patients undergoing ICSI, and follicular fluid samples were obtained from the leading follicle after ovarian stimulation. The corresponding embryos were evaluated and correlations with intrafollicular sRAGE concentrations were investigated using multi-adjusted regression analysis. We observed that high intrafollicular concentrations of sRAGE predicted poor embryo quality. Our findings suggest an association between high concentrations of intrafollicular sRAGE and poor embryo development following ovarian stimulation for ICSI.

Gender incidence of intracytoplasmic morphologically selected sperm injection-derived embryos: a prospective randomized study.

The aim of this prospective randomized study was to determine if the use of intracytoplasmic morphologically selected sperm injection (IMSI) is associated with gender incidence. Couples who underwent IVF-preimplantation genetic screening (PGS) cycles, as a result of advanced maternal age, were randomly allocated into two groups: intracytoplasmic sperm injection (ICSI; n=80) or intracytoplasmic morphologically selected sperm injection (IMSI; n=80). The incidences of genders were compared between ICSI- and IMSI-derived embryos. Considering all the biopsied embryos were characterized as normal for sex chromosome, the results showed that IMSI results in a significantly higher incidence of female embryos as compared with ICSI (65.1% versus 54.0%, respectively, P=0.0277). After analysing only euploid embryos for the eight selected chromosomes, a significantly higher incidence of XX embryos derived from IMSI was also observed compared with ICSI cycles (66.9% versus 52.5%, respectively, P=0.0322). This result was confirmed by logistic regression, which demonstrated a nearly 2-fold increase in euploid XX embryos derived from spermatozoa selected by high magnification (OR 1.83, 95% CI 1.05-3.35, P=0.032). A higher proportion of morphologically normal spermatozoa analysed under high magnification seem to carry the X chromosome. The aim of this study was to determine if the use of intracytoplasmic morphologically selected sperm injection (IMSI) is associated with gender incidence. Couples who underwent IVF with preimplantation genetic screening, as a result of advanced maternal age, were randomly allocated into two groups: intracytoplasmic sperm injection (ICSI; n=80) or intracytoplasmic morphologically selected sperm injection (IMSI; n=80). The incidences of genders were compared between ICSI- and IMSI-derived embryos. Our results showed that a significantly higher incidence of female embryos derived from IMSI compared with ICSI cycles (66.9% versus 52.5%, respectively, P=0.0322). This result was confirmed by logistic regression, which demonstrated a nearly 2-fold increase in female embryos derived from sperm selected by high magnification (OR 1.83, 95% CI 1.05-3.35, P=0.032). A higher proportion of morphologically normal spermatozoa analysed under high magnification seem to carry the X chromosome.

Varicocelectomy does not impact pregnancy outcomes following intracytoplasmic sperm injection procedures.

There are many studies in the literature suggesting an acquired, apparently progressive infertility due to varicocele. In fact, varicocelectomy has become the most commonly performed male infertility surgery. Assisted reproductive technologies such as intracytoplasmic sperm injection (ICSI) are also important for couples with male factor infertility associated with varicocele. Therefore, the aim of this study was to evaluate the effect of varicocelectomy on sperm quality and pregnancy rate with ICSI. Data were analyzed from 248 patients who had varicocele or underwent a previous varicocelectomy and were treated with ICSI between 2000 and 2008. Patients with varicocele were divided into two groups: men with clinical varicocele (group 1, n = 79) and men who underwent varicocelectomy before ICSI (group 2, n = 169). In all cases, female infertility was not detected. We evaluated and compared the two groups' semen characteristics as defined by the World Health Organization and Tygerberg's strict criteria: the female partner's age; the number of oocytes retrieved; and the fertilization, implantation, pregnancy, and miscarriage rates. We used the Wilcoxon signed rank test or the Mann-Whitney test for these analyses. No differences were detected in the age of the female partners between group 1 (33.0 ± 0.46 years) and group 2 (33.8 ± 0.38 years; P = .1872). Semen volume was higher in group 1 (3.3 ± 0.3 mL) than it was in group 2 (2.5 ± 0.14; P = .0043). No differences were detected between groups 1 and 2 with regard to sperm concentration (30.08 ± 4.01 million/mL and 24.1 ± 2.42 million/mL, respectively; P = .138), sperm motility (38.2% ± 2.69% and 38.7% ± 2.08%, respectively; P = .881), and morphology according to Tygerberg's strict criteria (2.6% ± 0.44% and 2.4% ± 0.37%, respectively; P = .7202). Also, no differences were detected in the number of oocytes retrieved between group 1 (14.8 ± 1.74) and group 2 (14.9 ± 1.04; P = .9515). The fertilization rate was higher in group 1 (73.2%) than it was in group 2 (64.9%; P = .0377); however, no differences were detected in the pregnancy rates (31.1% vs 30.9%; P = .9806), implantation rates (22.1% vs 17.3%; P = .5882), or miscarriage rates (21.7% vs 23.9%; P = .8401) between groups 1 and 2. Although a varicocelectomy should always be performed before assisted reproduction is pursued, this surgery does not increase pregnancy rates or decrease miscarriage rates following ICSI.

Prognostic value of triploid zygotes on intracytoplasmic sperm injection outcomes.

PURPOSE: To evaluate the prognostic significance of triploidy incidence on the outcomes of embryos derived from normally fertilized oocytes from the same cohort. METHODS: This study included 1500 ICSI cycles. Logistic regression models were used to study the influence of abnormal fertilization on the development and clinical outcomes of embryos derived from normally fertilized oocytes from the same cohort RESULTS: We observed a negative influence of the percentages of triploid zygotes on fertilization (75.2% and 56.8%, P < 0.0001), high-quality embryos (58.9% and 48.2%, P = 0.0001), pregnancy (34.1% and 28.2%, P = 0.0540) and implantation rates (20.0% and 13.3%, P = 0.0012). When the 3PN zygote rate was >25%, the percentages of normal fertilization, high-quality embryos and implantation rates were significantly lower than in the control group. CONCLUSIONS: We observed an approximately 50% lower risk of pregnancy and a 3.5-fold higher risk of miscarriage in cycles with a 3PN incidence of >25%.

Relationship between oocyte abnormal morphology and intracytoplasmic sperm injection outcomes: a meta-analysis.

The aim of this study was to perform a meta-analysis of the potential effects of oocyte morphological abnormalities on ICSI outcomes. Relevant original papers reporting on the relation between oocyte morphology and ICSI outcomes were identified by searching MEDLINE, EMBASE and the Cochrane Library. The main outcome measures were fertilisation rate and embryo quality. A meta-analysis was performed and Mantel-Haenszel pooled odd ratios (ORs) with 95% confidence intervals (CIs) were calculated to express the relation between the oocyte morphology and the ICSI outcomes. A total of 14 studies reporting 3688 ICSI cycles were included. Our meta-analysis demonstrates that the probability of an oocyte becoming fertilised is significantly reduced by the presence of large IPB (OR: 0.29, CI: 0.09-0.90), large PVS (OR: 0.86, CI: 0.74-0.99), refractile bodies (OR: 0.66, CI: 0.51-0.84) or vacuoles (OR: 0.59, CI: 0.42-0.83). No other investigated morphological abnormalities demonstrated significant relationships with ICSI outcomes. Our data demonstrate that the presence of large IPB, large PVS, refractile bodies or vacuoles is associated with decreased oocyte fertilisation. Our findings might be of importance for selecting embryos for replacement because the effects of oocyte abnormalities on implantation and pregnancy rates remain unclear.

Secretome of the preimplantation human embryo by bottom-up label-free proteomics.

A bottom-up label-free mass spectrometric proteomic strategy was used to analyse the protein profiles of the human embryonic secretome. Culture media samples used for embryonic culture of patients undergoing intracytoplasmic sperm injection cycles were selected as a test case for this exploratory proof-of-principle study. The media were stored after embryo transfer and then pooled into positive (n = 8) and negative (n = 8) implantation groups. The absolute quantitative bottom-up technique employed a multidimensional protein identification technology based on separation by nano-ultra-high pressure chromatography and identification via tandem nano-electrospray ionization mass spectrometry with data-independent scanning in a hydrid QqTOF mass spectrometer. By applying quantitative bottom-up proteomics, unique proteins were found exclusively in both the positive- and negative-implantation groups, which suggest that competent embryos express and secrete unique biomarker proteins into the surrounding culture medium. The selective monitoring of these possible secretome biomarkers could make viable procedures using single-embryo transfer.