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Research question Is there an influence of maternal lifestyle factors on the incidence of oocyte dimorphisms and the outcomes of intracytoplasmic sperm injection (ICSI) cycles? Design A total of 752 female patients undergoing ICSI cycle, in a private university-affiliated in vitro fertilization center, from January/2015 to December/2019, were included in this historical cohort study. Prior to start of controlled ovarian stimulation (COS), participants answered a questionnaire regarding cigarette smoking habit, consumption of items such as alcoholic beverages, refined sugar, artificial sweetener, soft drinks, fruits, legumes and vegetables, milk and dairy, and meats, and exercise frequency over the past 6 months. Oocyte morphology was evaluated before ICSI. The influence of maternal lifestyle factors on the incidence of oocyte dimorphisms and ICSI outcomes was evaluated by multivariate general linear models and generalized linear models, adjusted for potential confounders. The main outcome measures were the incidence of oocyte dimorphisms per cycle and clinical outcomes. Results Lifestyle factors and nutritional habits such as cigarette smoking, and the consumptions of alcohol, refined sugar, and artificial sweetener were positively associated with incidence of several oocyte dimorphisms and negatively associated with the response to COS and embryo development. Negative relationships were also observed between those habits and pregnancy outcomes, apart from miscarriage rate, in which positive relationships were observed. Significant negative dose-dependent relationships between those habits and implantation rate were noted. The intake of alcoholic beverages also showed inverse dose-dependent relationships with pregnancy and live-birth rates. Live-birth rate was also negatively associated with cigarette smoking, in a dose-dependent manner. Conclusions Bad maternal habits were associated with reduced oocyte quality and ICSI outcomes. Many of those associations were shown to be dose-dependent.

To investigate whether patients with a previous recombinant follicle stimulating hormone (rFSH)-stimulated cycle would have improved outcomes with rFSH + recombinant luteinizing hormone (rLH) stimulation in the following cycle. For the present retrospective case-control study, 228 cycles performed in 114 patients undergoing intracytoplasmic sperm injection (ICSI) between 2015 and 2018 in an in vitro fertilization (IVF) center were evaluated. Controlled ovarian stimulation (COS) was achieved with rFSH (Gonal-f, Serono, Geneva, Switzerland) in the first ICSI cycle (rFSH group), and with rFSH and rLH (Pergoveris, Merck Serono S.p.A, Bari, Italy) in the second cycle (rFSH + rLH group). The ICSI outcomes were compared among the groups. Higher estradiol levels, oocyte yield, day-3 high-quality embryos rate and implantation rate, and a lower miscarriage rate were observed in the rFSH + rLH group compared with the rFSH group. In patients 35 years old, the implantation rate was higher in the rFSH + rLH group compared with the rFSH group. In patients ≥ 35 years old, higher estradiol levels, oocyte yield, day-3 high-quality embryos rate, and implantation rate were observed in the rFSH + rLH group. In patients with ≤ 4 retrieved oocytes, oocyte yield, mature oocytes rate, normal cleavage speed, implantation rate, and miscarriage rate were improved in the rFSH + rLH group. In patients with ≥ 5 retrieved oocytes, higher estradiol levels, oocyte yield, and implantation rate were observed in the rFSH + rLH group. Ovarian stimulation with luteinizing hormone (LH) supplementation results in higher implantation rates, independent of maternal age and response to COS when compared with previous cycles stimulated with rFSH only. Improvements were also observed for ICSI outcomes and miscarriage after stratification by age and retrieved oocytes. Investigar se há algum efeito da suplementação com hormônio luteinizante (LH, na sigla em inglês) no regime com antagonista do hormônio liberador de gonadotropina (GnRH, na sigla em inglês) sobre os resultados dos ciclos consecutivos de injeção intracitoplasmática de espermatozoides (ICSI, na sigla em inglês). MéTODOS: Para o presente estudo retrospectivo de caso-controle, foram avaliados 228 ciclos de microinjeção intracitoplasmática de espermatozoides (ICSI, na sigla em inglês) realizados em 114 pacientes entre 2015 e 2018 em um centro privado de fertilização in vitro (FIV) afiliado a uma universidade. O estímulo ovariano controlado (EOC) foi feito com hormônio folículo- estimulante recombinante (rFSH, na sigla em inglês) (Gonal-f, Serono, Genebra, Suíça) no primeiro ciclo de ICSI (grupo rFSH), e com rFSH e rLH (Pergoveris, Merck Serono S.p.A, Bari, Itália) no segundo ciclo (grupo rFSH + rLH). Os desfechos dos ciclos de ICSI foram comparados entre os grupos. Níveis mais elevados de estradiol, de recuperação oocitária, taxa de embriões de alta qualidade no 3° dia e taxa de implantação, e menor taxa de aborto foram observados no grupo rFSH + rLH. Em pacientes 35 anos, a taxa de implantação foi maior no grupo rFSH + rLH em comparação com o grupo rFSH. Em pacientes com ≥ 35 anos, maiores níveis de estradiol, recuperação oocitária, a taxa de embriões de alta qualidade no 3° dia e a taxa de implantação foram observados no grupo rFSH + rLH. Em pacientes com baixa resposta ao EOC (≤ 4 oócitos recuperados), a recuperação oocitária, a taxa de oócitos maduros, a taxa de velocidade normal de clivagem, a taxa de implantação e a taxa de aborto foram melhoradas no grupo rFSH + rLH. Em pacientes com resposta normal ao EOC (≥ 5 oócitos recuperados), níveis mais elevados de estradiol, recuperação oocitária e taxa de implantação foram observados no grupo rFSH + rLH. CONCLUSãO: A estimulação ovariana com suplementação de LH resultou em taxas de implantação mais altas, independentemente da idade materna e da resposta ao EOC, em comparação com os ciclos anteriores estimulados apenas com rFSH. Melhorias também foram observadas nos resultados da ICSI e na taxa de aborto quando as pacientes foram estratificadas por idade e número de oócitos recuperados.

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.

Background Coronavirus disease 2019 (COVID‐19), which causes serious respiratory illnesses such as pneumonia and lung failure, was first reported in mid‐December 2019 in China and has spread around the world. In addition to causing serious respiratory illnesses such as pneumonia and lung failure, there have been conflicting reports about the presence of SARS‐CoV‐2 in the semen of patients who were previously diagnosed with COVID‐19 and possible implications for the male reproductive tract. Objective The goal for the present study was to review the current status of the literature concerning COVID‐19 and male reproduction. Material and methods An electronic literature search was done by using PubMed and Google Scholar databases. Relevant papers, concerning SARS‐COV‐2 and COVID‐19 and male reproduction, published between January 2020 and December 2020 were selected, analyzed and eventually included in the present literature review. Results SARS‐CoV‐2 may infect any cell type expressing angiotensin‐converting enzyme 2 (ACE2), including reproductive cells. Besides the presence of the SARS‐CoV‐2 receptor, the expression of host proteases, such as transmembrane serine protease 2 (TMPRSS2), is needed to cleave the viral S protein, allowing permanent fusion of the viral and host cell membranes. Here, we aimed to review the current status of the literature concerning COVID‐19 and male reproduction. The lack of co‐expression of ACE2 and TMPRSS2 in the testis suggests that sperm cells may not be at increased risk of viral entry and spread. However, the presence of orchitis in COVID‐19‐confirmed patients and compromised sex‐related hormonal balance among these patients intrigues reproductive medicine. Discussion SARS‐CoV‐2 may use alternate receptors to enter certain cell types, or the expression of ACE2 and TMPRSS2 may not be detected in healthy individuals. Conclusion COVID‐19 challenges all medical areas, including reproductive medicine. It is not yet clear what effects, if any, the COVID‐19 pandemic will have on male reproduction. Further research is needed to understand the long‐term impact of SARS‐CoV‐2 on male reproductive function.

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

The objective of this cross-sectional study was to investigate the impact of paternal age on the outcomes of intracytoplasmic sperm injection (ICSI) cycles at different values of maternal age. A total of 21,960 injected oocytes deriving from 3837 ICSI cycles performed between January 2014 and October 2020, performed in a private university-affiliated in vitro fertilization centre was included. The main effects of maternal and paternal age, as well as the effect of their product (interaction term) on embryo development and pregnancy outcomes were investigated considering the clustering of data. The coefficients for the interaction term were statistically significant for blastocyst development, top-quality blastocyst, implantation, pregnancy, miscarriage, and live-birth rates. For every 1-year increase in paternal age, the odds ratio of live-birth reduces by 1% in females aged 37 years, 1.6% in those aged 38 years, 2.4% in 39-year-old females, 5% in 42-year-old females and so on. An increase in the interaction term by 1 year decreases the pregnancy rate by 0.4% and live-birth rate by 0.8 and increases the miscarriage rate by 1.2%. The slopes of maternal age on blastulation, blastocyst quality, and implantation, pregnancy, miscarriage, and live-birth rate significantly changed (worsened) for every year increase in paternal age.

SummaryThe objective of this study was to investigate whether, in consecutive intracytoplasmic sperm injection (ICSI) cycles, embryonic development in an incubator with a time-lapse imaging (TLI) system is better than the previous one obtained in a benchtop incubator (G-185) with similar cultivation characteristics. The study was of a retrospective within-subject design, in which each cycle served as its own control. Data were obtained via the chart review of patients undergoing ICSI in a private university-affiliated in vitro fertilization (IVF) centre who fulfilled the following criteria: second ICSI attempt in which embryos were cultured in a TLI incubator system (TLI group, n = 71), preceded by a first ICSI attempt in which embryos were cultured in a benchtop incubator (Control group, n = 71). Embryonic development up to the fifth day of development, oocyte utilization rate (OUR; transferred embryos plus frozen embryos per total number of retrieved oocytes) and embryo utilization rate (EUR; transferred embryos plus frozen embryos per normally fertilized oocyte) were compared between the groups. There were significant differences in the day 2 non-cleavage rate, day 5 embryo rate, blastocyst development rate, frozen blastocyst rate, OUR, and EUR, in favour of the TLI group. Embryonic development, frozen blastocyst rate, OUR and EUR in the second ICSI cycle were significantly improved when the culture was performed in the EmbryoScope, compared with those rates obtained with culture in a G-185 in the first ICSI cycle of the same patients. The results may also lead to higher cumulative pregnancy outcomes following embryo thawing and transfer.

SummaryThe objective of this study was to investigate the effect of oocyte and sperm cryopreservation on donated eggs submitted to intracytoplasmic sperm injection (ICSI) cycles. Medical charts of 122 oocyte recipients undergoing 152 oocyte recipient ICSI cycles, from 2017 to 2018, in a private university-affiliated in vitro fertilization (IVF) centre, were reviewed in this historical cohort study. Cycles were divided into four groups according to the gamete status: the FO/FS Group, recipients in which fresh oocytes were injected with fresh sperm (n = 19); the FO/CrS Group, recipients in which fresh oocytes were injected with cryopreserved sperm (n = 14); the CrO/FS Group, recipients in which cryopreserved oocytes were injected with fresh sperm (n = 85); and the CrO/CrS Group, recipients in which cryopreserved oocytes were injected with cryopreserved sperm (n = 34). Generalized mixed models fit by restricted maximum likelihood, followed by Bonferroni post hoc test for the comparison of means amongst groups were used to investigate the effect of cryopreservation on recipient ICSI outcomes. The results were expressed as mean differences with 95% confidence intervals and P-values. The main outcome measure was the implantation rate. Normal day 3 cleavage speed, blastocyst development and implantation rates were significantly lower in the CrO/CrS Group compared with the FO/FS Group. In conclusion, embryo developmental competence and implantation potential were reduced when vitrified oocytes were injected with frozen sperm in an egg-sharing donation programme.

SummaryThe objective of this study was to investigate any effect of cryopreservation of donated eggs on laboratorial and clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles. This retrospective cohort study included 320 oocyte recipients undergoing 307 vitrified and 119 fresh oocyte recipient ICSI cycles, participating in an egg-sharing donation programme, from 2015 to 2018, in a private university-affiliated in vitro fertilization (IVF) centre. A review of donor and recipient ICSI cycles was charted. A general mixed models fit by restricted maximum likelihood, followed by Bonferroni post hoc test was used to compare the means between fresh and warm oocyte donation groups and investigate the effect of cryopreservation on recipient ICSI outcome. The main outcome measure was blastocyst development rates. Fertilization rate, high-quality embryo rates on days 2 and 3, normal cleavage speed rates on days 2 and 3, and blastocyst development rate were significantly higher for the fresh oocyte donation cycles compared with warmed oocyte donation cycles. In the egg-sharing donation programme, fertilization and embryo developmental competence were reduced when vitrified oocytes from infertile couples were used for ICSI compared with fresh oocytes.

OBJECTIVE The aim of this study was to investigate which factors contribute to the incidence of immature oocytes (germinal vesicle -GV- and metaphase I -MI-) and how they impact the intracytoplasmic sperm injection (ICSI) outcomes of sibling mature oocytes. METHODS Data from 3,920 cycles performed from June/2010 to August/2016 in a private university-affiliated IVF center were evaluated for the influence of controlled ovarian stimulation protocol (COS) on immature oocytes incidence and its effects on ICSI outcomes. RESULTS MI (p=0.004) and GV (p=0.029) number were negatively correlated with gonadotropin dose. Patients stimulated by rFSH had increased GV/oocyte rate in both GnRH agonists (p

To investigate whether the timing of rLH addition to rFSH during controlled ovarian stimulation (COS) impacts ovarian response and the outcomes of intracytoplasmic sperm injection (ICSI) cycles.Data of 1278 patients undergoing ICSI between 2015 and 2018, in a private university-affiliated IVF center were analyzed. Patients were divided into groups according to the timing of LH addition to the COS protocol: Group LH-start (n=323), in which LH was administered since day 1 of ovarian stimulation; and Group LH-mid (n=955), in which LH was administered concomitantly with gonadotropin releasing hormone (GnRH) antagonist. Data were also stratified according to female age and response to COS. The outcomes of COS and ICSI were compared between the groups.For the general group and in patients aged ≥ 35 years, higher blastocyst development rates were in Group LH-mid compared to Group LH-start. In patients with poor response to COS (POR), higher fertilization rate, blastocyst development rate and implantation rate were observed in Group LH-mid.rLH supplementation in POR patients may improve laboratorial and clinical outcomes when started in the mid-follicular phase, in GnRH antagonist ICSI cycles.

Background The paternal role in embryogenesis is much more than providing a haploid genome. The spermatozoa have crucial roles related to embryogenesis' epigenetic regulation, fusion of gametes, and cleavage, which may influence intracytoplasmic sperm injection outcomes. Objectives To study whether paternal age, length of ejaculatory abstinence and semen parameters influence the outcomes of intracytoplasmic sperm injection in the oocyte recipients from egg-sharing donation cycles. Materials and methods This historical cohort study involved 427 oocyte recipient intracytoplasmic sperm injection cycles, from 321 patients undergoing egg-sharing donation cycles from January 2015 to May 2017, in a private in vitro fertilization center. Electronic medical records of donors and recipients intracytoplasmic sperm injection cycles were reviewed. The impact of male age, length of ejaculatory abstinence, and semen parameters on recipients' intracytoplasmic sperm injection outcomes were investigated using general mixed models. Implantation, pregnancy, and live birth rates were the primary outcome measures. Results Paternal age negatively affected fertilization, day 3 top-quality embryos, day 3 normal embryo cleavage speed, blastocyst formation, high-quality blastocysts, and implantation, and was correlated with diminished pregnancy and live birth rates. Length of ejaculatory abstinence negatively influenced the rates of day 3 top-quality embryos, normal embryo development on day 3, blastocyst development, and implantation. Sperm count positively affected the rates of fertilization, blastocyst development, and implantation. The percentage of progressive sperm motility positively affected day 3 normal embryo development rate and implantation rate. Total motile sperm count positively affected blastocyst development rate and implantation rate. Discussion and conclusion Additional tracking of the influence of male characteristics on intracytoplasmic sperm injection outcomes should be stimulated. Paternal age is not manageable, but reducing length of ejaculatory abstinence could be an alternative approach to improve the outcomes of intracytoplasmic sperm injection.

The objective of this study was to investigate the impact of male age, semen quality and days of ejaculatory abstinence on embryo morphokinetics. A total of 1,220 zygotes obtained from 139 couples in a private in vitro fertilisation centre were analysed. The timing of specific events from the point of insemination, such as timings to pronuclei appearance and fading, to two, three, four, five, six, seven and eight cells and to blastulation were recorded. Multivariate linear regression analysis was used to evaluate the influence of paternal factors on embryo morphokinetic events. Paternal age was positively correlated with delayed cell cleavage and blastulation, and negatively associated with implantation rate, and clinical pregnancy and live-birth chances. The ejaculatory abstinence was inversely correlated with the implantation rate. Inverse relationships were observed between semen parameters (sperm count, progressive sperm motility, total motile sperm count and morphology) and the timing of specific events during embryo development. Sperm morphology was also positively associated with implantation rate and pregnancy and live-birth chances. Increased paternal age and ejaculatory abstinence, and poor semen quality correlate with delayed cell cleavage and blastulation and negatively impact intracytoplasmic sperm injection outcomes.

Study question Are the morphological parameters and development of in vitro cultured embryos, and intracytoplasmic sperm injection (ICSI) outcomes influenced by maternal and paternal ageing? Summary answer The slopes of maternal age on blastulation, blastocyst quality, and implantation, pregnancy and miscarriage rates significantly changed (worsened) for every year increase in paternal age. What is known already Due to the vast literature demonstrating that female age interferes with intracytoplasmic sperm injection (ICSI) outcomes, there is an imposition, in numerous countries, regarding maternal age limit for assisted reproduction. Despite several studies have underscored the negative impact of paternal age and lifestyle factors on reproductive health, the influence of paternal age on ICSI outcomes is still a matter of debate. The aim of this study was to investigate if the effect of paternal age on embryo development differs at different values of maternal age, thus creating a rationale for the data to reach physicians, patients, and public health recommendations. Study design, size, duration This historical cohort study included 3837 couples undergoing their first ICSI cycle from January/2014 to October/2020. A total of 21960 oocytes were injected and embryos were evaluated until day 5 of development. The main effects of maternal and paternal ages, as well as the effect of their product (interaction term) on embryo growth and development, and on pregnancy outcomes were investigated taking into account clustering of data (multiple embryos per cycle), using generalized mixed models. Participants/materials, setting, methods The study was performed in a private university-affiliated in vitro fertilization center. Zygotes were morphologically evaluated 17h post ICSI. For days 2 and 3 of development, the number of blastomeres, blastomere symmetry, percentage of fragmentation and presence of multinucleation were recorded. On day 5 of development, successful blastulation, and inner cell mass and trophectoderm qualities were recorded. Pregnancy was calculated per transfer, and miscarriage was defined as pregnancy loss before 20 weeks gestation. Main results and the role of chance The coefficients for the interaction term were statistically significant for blastocyst development (B: - 0.005, OR: 0.995, CI: 0.994 – 0.996, p Limitations, reasons for caution The retrospective and monocentric nature of the study are its major limitations. Wider implications of the findings Our results underscore the importance of both maternal and paternal ages for blastulation and successful pregnancy. Main effects of paternal and maternal ages should no longer be interpreted as the relationship between each independent variable and a given outcome, but rather be conditional on the values of the interaction term. Trial registration number Not applicable

Study question Does the impact of sperm DNA fragmentation (SDF) on Intracytoplasmic sperm injection (ICSI) outcomes depend on the presence of oocyte dimorphisms? Summary answer There is a significant influence of oocyte quality on the impact of SDF on pregnancy outcomes. What is known already Sperm DNA fragmentation has been associated with ICSI outcomes. DNA damage is commonly encountered in human spermatozoa and it has been widely accepted that the oocyte assumes responsibility for the repair and remodelling of both the maternal and paternal genomes during the oocyte-embryo transition. Indeed, spermatozoa with DNA damage can fertilise oocytes and still lead to embryo development due to the oocyte DNA repair capacity. Considering the vital role played by the oocyte in the developmental process, it could be hypostatised that the oocyte quality, translated as oocyte morphology, would influence the machinery responsible for sperm DNA repair after fertilization. Study design, size, duration This cohort study included 3,035 oocytes from 525 patients undergoing ICSI cycles in a university-affiliated IVF-center, between June/2016 and July/2019. Oocytes were split into groups according to the SDF index of the sample used for ICSI: low-fragmentation ( Participants/materials, setting, methods Data was evaluated using generalized linear models (GZLM) followed by Bonferroni post hoc. The results are expressed as mean ± standard error for continuous variables or percentages for dichotomous variables, and p-values. The sample size calculation suggested that a sample of at least 504 subjects had 95% power to detect a 20% effect with a significance level of 5% (α). The study was performed in a private university–affiliated in vitro fertilization (IVF) center. Main results and the role of chance The association of both factors: the presence of oocyte dimorphisms (dark cytoplasm, vacuoles in the ooplasm, and resistant membrane) and high SDF index resulted in the lowest fertilization rate among groups, while oocytes free of these dimorphisms injected with samples with Limitations, reasons for caution The retrospective nature is a limitation. It could be argued that using samples with high SDF index does not necessarily mean that a sperm cell with a fragmented DNA was injected, however, the higher the SDF index, the higher the chance of selecting one with fragmented DNA. Wider implications of the findings The findings presented here highlight the crucial role of male and female factors when facing assisted reproduction. The association of low oocyte quality and high SDF index may lead to impaired results. As the oocyte defect cannot be modified, in vivo upgrading of spermatozoa before the treatment should be encouraged. Trial registration number Not applicable

To study the implications of sperm DNA fragmentation (SDF) in intracytoplasmic sperm injection cycles for non-male factor infertility.Prospective cohort study.Private university-affiliated IVF center.Data from 475 cycles performed from June 2016 to June 2017.Cycles were divided according to SDF rate into two groups:30% SDF (n = 433) and ≥30% SDF (n = 42). Laboratory and clinical outcomes were compared between groups by generalized linear models adjusted for potential confounders.Embryo quality and miscarriage rates.Fertilization rate was similar between groups (≥30% SDF, 85.28% ± 1.06% vs.30% SDF, 90.68% ± 3.61%). Significantly lower rates of normal cleavage speed (≥30% SDF, 61.12% ± 4.21% vs.30% SDF, 72.53% ± 1.24%), high-quality embryos at day 3 (≥30% SDF, 23.07% ± 5.56% vs.30% SDF, 36.41% ± 1.53%), blastocyst formation (≥30% SDF, 39.09% ± 2.73% vs.30% SDF, 58.83% ± 7.59%), blastocyst quality (≥30% SDF, 11.97% ± 1.22% vs.30% SDF, 30.09% ± 2.39%), and implantation (33.24% ± 1.66% vs.30% SDF, 46.40% ± 4.61%) were observed in cycles with higher SDF, despite similar pregnancy rates (≥30% SDF, 30.40% vs.30% SDF, 32.40%). A 2.5-fold miscarriage rate was observed in cycles with an SDF above the established cutoff (≥30% SDF, 42.8% vs.30% SDF, 16.8%).Higher SDF is correlated with poor embryo development, lower implantation rate, and higher miscarriage rate in non-male factor infertility intracytoplasmic sperm injection cycles. Since defects in sperm may be hidden, the SDF test can bring additional information to the sperm quality evaluation of men with unknown infertility history.

Research question Are the outcomes of (i) surgical sperm retrieval (SSR) and (ii) intracytoplasmic sperm injection (ICSI) influenced by the obstructive interval (time elapsed since vasectomy)? Design Medical records from 148 patients (194 cycles) with secondary azoospermia due to vasectomy, who presented for percutaneous epididymal sperm aspiration (PESA) and ICSI in a private university-affiliated IVF centre, from January 2012 to February 2017, were analysed in this historical cohort study. The obstructive interval was recorded for each couple, and its influences on the outcomes of SSR and ICSI treatment were investigated using general mixed models with adjustment for potential confounders. Clinical pregnancy rate was the main outcome measure. Results The obstructive interval was negatively correlated with the presence of spermatozoa (β = –0.032, P = 0.009) and motile spermatozoa (β = –0.031, P = 0.010) during PESA. The need to convert to testicular sperm aspiration was significantly influenced by the obstructive interval (β = 0.012, P = 0.003). The blastocyst development rate on day 5 was inversely correlated with the obstructive interval (β = –0.011, P = 0.014). Implantation and clinical pregnancy rates were negatively influenced by the obstructive interval (β = –1.107, P = 0.039 and β = –0.016, P = 0.031, respectively). The receiver operating characteristic curve analysis demonstrated that the obstructive interval has a predictive value on the achievement of clinical pregnancy (area under the curve = 0.667, P = 0.001, Youden index 0.3385, associated criterion >17 years). Conclusions Men undertaking vasectomy should be made aware of the long-term effects and their implications for future reproductive treatment.

PURPOSE: To study the association between body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR), and response to controlled ovarian stimulation, semen quality, and intracytoplasmic sperm injection (ICSI) outcomes. METHODS: This prospective observational study included data from 402 couples undergoing ICSI between January/2010–September/2016. Participants had their weight, height, waist, and hip measured before ICSI treatment. RESULTS: Gonadotropin dose was positively associated with maternal WHR. Maternal WC and WHR negatively influenced the estradiol level, number of follicles, oocytes, and mature oocytes. Maternal BMI negatively influenced oocyte yield. Semen volume was negatively influenced by male BMI. Sperm concentration was negatively associated with WHR and WHtR. Progressive sperm motility was negatively influenced by all anthropometric measures (AMs). The TMSC was negatively affected by all AM but WC. Significant differences were observed in all ICSI outcomes except pregnancy and miscarriage rates, favoring women with AM below the cut off values compared to women above the cut off. It was observed that men with AM below the cut off values showed significantly higher rates of fertilization, embryo and blastocyst development, implantation, and pregnancy than men above the cut off. When combined maternal and paternal AM were analyzed, significant differences were observed in fertilization, embryo and blastocyst development and implantation rates, favoring couples with normal BMI, WC, and WHR. CONCLUSIONS: Lower AMs have a protective effect on fertility. Awareness and counseling on how to minimize its impact in both partners could improve ICSI outcomes.

OBJECTIVE To evaluate if ovarian response to controlled ovarian stimulation (COS) and intracytoplasmic sperm injection (ICSI) outcomes are improved by the use of dual trigger (gonadotropin-releasing hormone (GnRH) agonists plus recombinant human chorionic gonadotropin (r-hCG)) in patients with previous cycles triggered with r-hCG. METHODS This case-control study included 88 matched cycles performed in 88 patients, which had the first ICSI cycle triggered with r-hCG (n=44), and the following ICSI cycle with dual trigger (n=44). We compared the cycle outcomes between the groups. In a second case-control within-subject analyses, we compared the ICSI outcomes between patients which had the first ICSI cycle triggered with r-hCG only (n=18), and the following ICSI cycle with dual trigger (n=18) or r-hCG only (n=18). RESULTS Upon investigating repeated cycles (r-hCG only vs. dual trigger), we found higher oocyte yield and mature oocyte rates, lower immature oocyte rates, higher fertilization rates, and higher blastocyst development rates; and higher rates of cycles with embryos transferred and implantation in the dual trigger cycle. CONCLUSIONS The dual trigger regimen is a more effective approach than r-hCG trigger in patients with a previous r-hCG triggered ICSI cycle, yielding improved response to COS, and better laboratorial and clinical outcomes.

Objective To study the impact of sperm DNA fragmentation (SDF) on clinical outcomes of assisted reproductive technology in women with different age ranges. Design Historical cohort study. Setting Private university–affiliated in vitro fertilization center. Patient(s) Five hundred forty couples undergoing intracytoplasmic sperm injection cycles. Intervention(s) Cycles were split into three groups according to maternal age: ≤36 years old (n = 285), 37–40 years old (n = 147), and >40 years old (n = 108). Semen samples were evaluated for SDF using the Sperm Chromatin Dispersion test and, for each age group, the cycles were subdivided according to SDF index: low fragmentation index ( Main Outcome Measure(s) Implantation, pregnancy, and miscarriage rates. Result(s) For younger patients (≤36 years old) and those between 37 and 40 years of age, no significant differences were noted in laboratory and clinical outcomes for cycles with 40 years of age, significantly lower high-quality day-3 embryos (54.4% vs. 33.1% and blastocyst development rates (49.6% vs. 30.2%), lower pregnancy (20.0% vs. 7.7%) and implantation rates (19.7% vs. 11.9%), and increased miscarriage rate (12.5% vs. 100.0%) were observed for cycles with ≥30% SDF compared with Conclusion(s) Older oocytes, when injected with sperm derived from samples with high SDF index, develop into embryos of poor quality that lead consequently to lower implantation and pregnancy rates and higher miscarriage rates, in intracytoplasmic sperm injection cycles from women with advanced maternal age.

Objective To investigate the predictive factors for successful pregnancy in oocyte recipient ICSI cycles in an egg-sharing donation program. Methods Analysed data were obtained via chart review of 1505 vitrified oocytes donated from 268 patients to 225 oocyte recipients, undergoing 307 ICSI cycles. Patients were participating in an egg-sharing donation program between January 2015 and May 2017. Adjusted generalised linear models were used to investigate the impact of oocyte donor and recipient characteristics on recipients’ pregnancy achievement. Results Implantation rate in the oocyte donor was highly correlated with pregnancy achievement in the oocyte recipient’s cycles (ExpB: 1.181, CI: 1.138-1.226, p

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.

The objective of this study was to investigate the effects of low-dose hCG supplementation on ICSI outcomes and controlled ovarian stimulation (COS) cost. Three hundred and thirty patients undergoing ICSI were split into groups according to the COS protocol: (i) control group (n = 178), including patients undergoing conventional COS treatment; and (ii) low-dose hCG group (n = 152), including patients undergoing COS with low-dose hCG supplementation. Lower mean total doses of FSH administered and higher mean oestradiol level and mature oocyte rates were observed in the low-dose hCG group. A significantly higher fertilization rate, high-quality embryo rate and blastocyst formation rate were observed in the low-dose hCG group as compared to the control group. The miscarriage rate was significantly higher in the control group compared to the low-dose hCG group. A significantly lower incidence of OHSS was observed in the low-dose hCG group. There was also a significantly lower gonadotropin cost in the low-dose hCG group as compared to the control group ($1235.0 ± 239.0×$1763.0 ± 405.3, p 0.001). The concomitant use of low-dose hCG and FSH results in a lower abortion rate and increased number of mature oocytes retrieved, as well as improved oocyte quality, embryo quality and blastocyst formation and reduced FSH requirements.

The couple from this clinical case consisted of a 55 years old male with an obstructive interval of 25 years post vasectomy, and a 38 years old female partner. Both partners had normal results in infertility workup. Five mature oocytes were injected with motile spermatozoa showing morphological alterations, obtained by percutaneous epididymal sperm aspiration. Four oocytes fertilized, and three embryos were transferred with assisted hatching on day three of development, of which one was a high-quality embryo. A clinical pregnancy was confirmed by the detection of two gestational sacs with foetal heartbeats. Pregnancy was ongoing during the submission of this manuscript. The use of ICSI with PESA/TESA should be considered as a feasible alternative when vasectomy reversal fails in vasectomized men wishing to father again. Case reports like this may inspire the counseling of couples that have suffered from a previous vasectomy reversal failure and support the recommendation of ICSI with PESA treatment, which could allow those couples to have their own children, even in the presence of advanced parental age.

Objective To study whether embryos derived from oocytes presenting a smooth endoplasmic reticulum cluster (SERC) are less likely to develop into blastocysts and implant. Design Transversal study. Setting Private university-affiliated in vitro fertilization (IVF) center. Patient(s) Total of 7,609 oocytes obtained from 743 intracytoplasmic sperm injection (ICSI) cycles. Intervention(s) Oocytes split between the SERC-positive cycles (with at least one SERC-positive oocyte) and the SERC-negative cycles (only oocytes free of SERC). Main Outcome Measure(s) Embryo implantation. Result(s) A statistically significantly higher mean number of follicles (24.0 ± 10.5 vs. 19.6 ± 10.5), retrieved oocytes (17.8 ± 8.3 vs. 14.3 ± 8.0), and mature oocytes (13.5 ± 6.2 vs. 10.6 ± 5.9) were observed in the SERC-positive cycles as compared with SERC-negative cycles. The implantation rate was statistically significantly lower in SERC-positive cycles as compared with SERC-negative cycles (14.8% vs. 25.6%; odds ratio 0.61; 95% confidence interval, 0.44–0.86). When only cycles with in which none (0) or all the blastocysts transferred had implanted (100%) were analyzed, the mean implantation rate per transferred blastocyst in the SERC-negative group was 20.5%; no blastocysts derived from SERC-positive oocytes implanted. Conclusion(s) The occurrence of SERC impairs embryo implantation. Careful oocyte observation that takes into account the presence of SERC should be part of embryo selection strategy before transfer.

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 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.

Objective To identify potential microRNA (miRNA) biomarkers of poor, normal and hyperresponse to controlled ovarian stimulation (COS). Methods In the present study, we assessed 40 serum samples from patients undergoing COS. We used ten samples to standardize miRNAs detection in the serum. The remaining 30 samples were split into three groups depending on the patient's response to COS: poor response (PR group, n=10), normal response (NR group, n=10), and hyperresponse (HR group, n=10). Aberrantly expressed miRNAs were identified using a large-scale expression analysis platform. Gene set enrichment analysis was performed to assess the biological processes potentially modulated by the identified miRNAs. Results Twenty-two miRNAs were detected only in the PR or HR groups when compared with the NR group. From those, 11 presented poor dissociation curves and were excluded from further analysis. A bioinformatics analysis revealed that the selected 11 miRNAs target several genes involved in GnRH, estrogen and prolactin signaling, oocyte maturation, female pregnancy, and meiosis. Conclusion The large-scale analysis of miRNA expression identified distinct miRNA profiles for poor and hyperresponse to COS, which potentially modulate key processes for human assisted reproduction. All evidence suggests that the serum microRNA profiling may discriminate patients who will respond in an exacerbated manner from those who will respond insufficiently to COS. Further studies may validate these miRNAs, enabling the individualization of treatment and more successful outcomes.

Objective To study which factors affect perinatal outcomes in intracytoplasmic sperm injection (ICSI) cycles. Methods Data was obtained from 402 live births born to 307 patients undergoing ICSI cycles in a private university-affiliated IVF center between Jan/2014 and Dec/2015. The influences of the cycles' characteristics on the number of gestational weeks to livebirth (GW), baby birth weight (BW), and baby birth length (BL) were evaluated by linear regression models, adjusted for maternal age and body mass index, number of transferred embryos, number of gestational sacs, and number of born infants. In a subsequent analysis, GW, BW and baby sex were utilized for cycle classification into the groups Appropriate for gestational age (AGA n=256) and Small for gestational age (SGA n=146), which were compared by general linear models adjusted for the same confounder variables. Results The number of follicles (β=-0.069 p=0.018) and retrieved oocytes (β=-0.087 p=0.049) were negatively correlated with BL. The endometrial thickness was positively correlated with GW (β=0.198 p=0.003) and BW (β=28.351 p=0.044). When each baby was classified into AGA and SGA groups, it was observed that SGA babies were derived from cycles with higher estradiol levels at hCG day (SGA: 3897.01±550.35 vs. AGA: 2324.78±101.86 p=0.006) and higher number of retrieved oocytes (SGA: 16.70±1.78 vs. AGA: 12.92±0.42 p=0.042). The endometrial thickness was significantly lower in the SGA group (SGA: 10.2±0.23 vs. AGA: 11.68±0.17 vs. p=0.029). Conclusion Higher ovarian response to stimulation and suboptimal endometrial development are associated with adverse perinatal outcomes in ICSI cycles.

Objective To describe the cases of preimplantation genetic testing for monogenic diseases (PGT-M) in fertile couples who had undergone intracytoplasmic sperm injection (ICSI) cycles in a Brazilian in vitro fertilisation (IVF) centre and determine whether these cases were different from those reported from the European Society of Human Reproduction and Embryology (ESHRE). Methods This retrospective collection included data obtained from ICSI-PGT-M cycles between 2011 and 2016. The disease indication, number of biopsied embryos, biopsy stage, diagnosed and affected embryos, and cycles with embryo to transfer as well as implantation, pregnancy and miscarriage rates were analysed and compared to cycles without genetic diagnosis (PGT) and with ESHRE PGD Consortium collection XIV-XV. Results From 5,070 cycles performed, 72 had indications for PGT-M. The most common time for biopsy was cleavage-stage; 93% of the embryos had a diagnostic result, 59.4% of which were genetically transferable, resulting in 68% of the cycles with transferred embryos, a 22.1% implantation rate, and a 28.6% pregnancy rate. No differences in clinical outcomes of cycles with PGT-M or without PGT were observed. The day of biopsy and diagnostic success as well as implantation, pregnancy and miscarriage rates were similar to ESHRE collection. Conclusions Although the proportion of cases with PGT-M was low, its efficacy was similar to what was reported in the European collection and represents a viable alternative for families at risk of transmitting a genetic disorder to their offspring. The main difference between our and ESHRE collection were the disease indications, which reflected the admixed, multi-ethnic Brazilian population.

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.

Objective: The novel coronavirus (Covid-19) outbreak led to a public health emergency of international concern, putting health organizations on alert World authorities implemented suppression plans to control community spread, including restrictions to non-urgent medical care Assisted reproduction centers had to adapt to these restrictions The infertility diagnosis and reproductive treatments possess an inherent psychological burden This associated with the uncertainty of the consequences of the passage of time in the prognosis of treatments may impact on patient’s psychological health The goal for the present study was to investigate whether women seeking fertility care have different perception concerning the impact of Covid-19 on the motherhood plan than a target population? Design: Prospective randomized study Materials and Methods: From 22/April/2020 to 25/may/2020, a survey through an online-platform was conducted Participants were randomized by age in one of the two groups: ART-GROUP (n=92), including patients seeking for fertility treatment, but still didn’t start their cycles or INTERESTED-GROUP (n=92), including participants interested in the subject, who accessed the website of a university-affiliated IVF-center Participants in the ART-GROUP were invited via e-mail, with a cover-letter outlining the survey and a link to access it Participants in the INTERESTED-GROUP accessed the questionnaire via website Information on demographic data and their perceptions in face of the COVID-19 pandemics and the motherhood plan was collected Women were asked: (i) How do you see the possibility of becoming pregnant after the beginning of the COVID-19 pandemic? (ii) How long do you think that suppression strategies will last? and (iii) Did you postpone your plans to become pregnant? If yes, why? Results: Most patients in the ART-GROUP were married or in a common-law relationship (83 6%), while a half of women in the INTERESTED-GROUP were in the same situation (50 0% p