Vertical transmission of maternal DNA through extracellular vesicles associates with altered embryo bioenergetics during the periconception period.

The transmission of DNA through extracellular vesicles (EVs) represents a novel genetic material transfer mechanism that may impact genome evolution and tumorigenesis. We aimed to investigate the potential for vertical DNA transmission within maternal endometrial EVs to the pre-implantation embryo and describe any effect on embryo bioenergetics. We discovered that the human endometrium secretes all three general subtypes of EV - apoptotic bodies (ABs), microvesicles (MVs), and exosomes (EXOs) - into the human endometrial fluid (EF) within the uterine cavity. EVs become uniformly secreted into the EF during the menstrual cycle, with the proportion of different EV populations remaining constant; however, MVs contain significantly higher levels of mitochondrial (mt)DNA than ABs or EXOs. During the window of implantation, MVs contain an eleven-fold higher level of mtDNA when compared to cells-of-origin within the receptive endometrium, which possesses a lower mtDNA content and displays the upregulated expression of mitophagy-related genes. Furthermore, we demonstrate the internalization of EV-derived nuclear-encoded (n)DNA/mtDNA by trophoblast cells of murine embryos, which associates with a reduction in mitochondrial respiration and ATP production. These findings suggest that the maternal endometrium suffers a reduction in mtDNA content during the preconceptional period, that nDNA/mtDNA become packaged into secreted EVs that the embryo uptakes, and that the transfer of DNA to the embryo within EVs occurs alongside the modulation of bioenergetics during implantation.

Endometrial receptivity tests in reproduction: a SWOT analysis.

Endometrial receptivity and its management in assisted reproduction is now a significant focus of research interest. Endometrial receptivity tests, which analyze different panels of gene expression, are usually offered in fertility clinics to determine the women's individual 'window of implantation', providing a personalized timing for embryo transfer. However, there are still no definite indications on whether its inclusion in the study of the infertile couple or the study of patients with repeated implantation failure is essential.

The step-up protocol increases clinical pregnancy rates compared with the step-down in patients with unexplained infertility. A randomized controlled trial.

OBJECTIVE: Unexplained infertility is a relevant indication for controlled ovarian stimulation associated to intrauterine insemination. The "step-up" and "step-down" gonadotropin-based protocols were designed to reduce multiple pregnancy and ovarian hyperstimulation syndrome in polycystic ovary syndrome patients, but there is no related evidence in normoovulatory women undergoing intrauterine insemination. Our aim was to compare the efficacy and safety of both protocols with intrauterine insemination in unexplained infertility patients. METHODS: Randomized clinical trial including 145 women with unexplained infertility randomly following the step-up (n=73) or step-down (n=72) protocol. In the step-up group, patients started on day 3 of a spontaneous cycle administrating recombinant FSH 75IU sc/day, increasing it to 150IU if no response after 7 days. In the step-down, patients started administrating 150IU sc/day, constantly decreasing it to 75IU after 5 days. Recombinant hCG was administered when a follicle reached ≥18mm diameter. RESULTS: Clinical pregnancy rate was higher in the step-up group than in the step-down (20.5% vs . 8.3%; p =0.037). Significant differences between step-up and step-down protocols were found regarding days of rFSH administration (8.83±4.01% vs . 7.42±2.18%; p =0.001) and cancellation rate due to hyper response (8.21% vs . 25%; p =0.05). No differences were detected in miscarriage rates, multiple pregnancy rates/cycle and hyper stimulation syndrome incidence. CONCLUSIONS: The step-up protocol is longer-lasting but more effective obtaining pregnancies than the step-down in patients with unexplained infertility undergoing intrauterine insemination. This effect could be explained by lower cancellation rates due to ovarian hyper response than the step-down protocol, with no differences in ovarian hyper stimulation syndrome incidence.

The effect of short term exposure to outdoor air pollution on fertility.

BACKGROUND: There is evidence to suggest that long term exposure to air pollution could be associated with decreased levels of fertility, although there is controversy as to how short term exposure may compromise fertility in IVF patients and what windows of exposure during the IVF process patients could be most vulnerable. METHODS: This prospective cohort study aimed to evaluate the impact of acute exposure that air pollution have on reproductive outcomes in different moments of the IVF process. Women undergoing IVF living in Barcelona were recruited. Individual air pollution exposures were modelled at their home address 15 and 3 days before embryo transfer (15D and 3D, respectively), the same day of transfer (D0), and 7 days after (D7). The pollutants modelled were: PM2.5 [particulate matter (PM) ≤2.5 µm], PMcoarse (PM between 2.5 and 10µm), PM10 (PM≤10 µm), PM2.5 abs, and NO2 and NOx. Outcomes were analyzed using multi-level regression models, with adjustment for co-pollutants and confouding factors. Two sensitivity analyses were performed. First, the model was adjusted for subacute exposure (received 15 days before ET). The second analysis was based on the first transfer performed on each patient aiming to exclude patients who failed previous transfers. RESULTS: One hundred ninety-four women were recruited, contributing with data for 486 embryo transfers. Acute and subacute exposure to PMs showed a tendency in increasing miscarriage rate and reducing clinical pregnancy rate, although results were not statistically significant. The first sensitivity analysis, showed a significant risk of miscarriage for PM2.5 exposure on 3D after adjusting for subacute exposure, and an increased risk of achieving no pregnancy for PM2.5, PMcoarse and PM10 on 3D. The second sensitivity analysis showed a significant risk of miscarriage for PM2.5 exposure on 3D, and a significant risk of achieving no pregnancy for PM2.5, PMcoarse and PM10 particularly on 3D. No association was observed for nitrogen dioxides on reproductive outcomes. CONCLUSIONS: Exposure to particulate matter has a negative impact on reproductive outcomes in IVF patients. Subacute exposure seems to increase the harmful effect of the acute exposure on miscarriage and pregnancy rates. Nitrogen dioxides do not modify significantly the reproductive success.

Lower risk of adverse perinatal outcomes in natural versus artificial frozen-thawed embryo transfer cycles: a systematic review and meta-analysis.

This systematic review of literature and meta-analysis of observational studies reports on perinatal outcomes after frozen embryo transfer (FET). The aim was to determine whether natural cycle frozen embryo transfer (NC-FET) in singleton pregnancies conceived after IVF decreased the risk of adverse perinatal outcomes compared with artificial cycle frozen embryo transfer (AC-FET). Thirteen cohort studies, including 93,201 cycles, met the inclusion criteria. NC-FET was associated with a lower risk of hypertensive disorders in pregnancy (HDP) (RR 0.61, 95% CI 0.50 to 0.73), preeclampsia (RR 0.47, 95% CI 0.42 to 0.53), large for gestational age (LGA) (RR 0.93, 95% CI 0.90 to 0.96) and macrosomia (RR 0.82, 95% CI 0.69 to 0.97) compared with AC-FET. No significant difference was found in the risk of gestational hypertension and small for gestational age. Secondary outcomes assessed were the risk of preterm birth (RR 0.83, 95% CI 0.79 to 0.88); post-term birth (RR 0.48, 95% CI 0.29 to 0.80); low birth weight (RR 0.84, 95% CI 0.80 to 0.89); caesarean section (RR 0.84, 95% CI 0.77 to 0.91); postpartum haemorrhage (RR 0.39, 95% CI 0.35 to 0.45); placental abruption (RR 0.61, 95% CI 0.38 to 0.98); and placenta accreta (RR 0.18, 95% CI 0.10 to 0.33). All were significantly lower with NC-FET compared with AC-FET. In assessing safety, NC-FET significantly decreased the risk of HDP, preeclampsia, LGA, macrosomia, preterm birth, post-term birth, low birth weight, caesarean section, postpartum haemorrhage, placental abruption and placenta accreta. Further randomized controlled trials addressing the effect of NC-FET and AC-FET on maternal and perinatal outcomes are warranted. Clinicians should carefully monitor pregnancies achieved by FET in artificial cycles prenatally, during labour and postnatally.

Laparoscopic Roux-en-Y gastric bypass versus laparoscopic sleeve gastrectomy for 5-year hypertension remission in obese patients: a systematic review and meta-analysis.

: Controversial results exist on mid-term effects of Roux-en-Y gastric bypass and sleeve gastrectomy on hypertension remission. The aim of the present systematic review was to study 5-year hypertension remission after both procedures. One-year hypertension remission and SBP and DBP pressure change at 1 and 5 years after both surgical techniques were also evaluated. We searched MEDLINE, EMBASE and The Cochrane Central Register of Controlled Trials (CENTRAL). Thirty-two articles were included (six randomized controlled trials, 18 cohort and eight case-control studies). The proportion of patients with hypertension remission was greater for those treated with gastric bypass compared with sleeve gastrectomy at 5 years (RR = 1.26, 95% CI = 1.07-1.48) and 1 year (RR = 1.14, 95% CI = 1.06-1.21). Gastric bypass and sleeve gastrectomy did not differ in terms of SBP or DBP change. Patients treated with gastric bypass present a higher hypertension remission rate at 1 and 5 years.

Second-generation preimplantation genetic testing for aneuploidy in assisted reproduction: a SWOT analysis.

Second-generation preimplantation genetic testing for aneuploidy (PGT-A 2.0) in patients with an unfavourable reproductive and IVF prognosis is becoming common practice, with the aim of improving reproductive outcomes. However, there is still no clear evidence on the possible advantages and drawbacks with regard to this procedure. In this discussion paper, based on a SWOT (strengths, weaknesses, opportunities, threats) analysis, the different aspects of this strategy are evaluated. Current evidence suggests that PGT-A 2.0 should not at present have an indiscriminate application, but it might be indicated in cases in which the risk of aneuploidy is increased.

Endometriosis does not affect live birth rates of patients submitted to assisted reproduction techniques: analysis of the Latin American Network Registry database from 1995 to 2011.

PURPOSE: The purpose of this study is to compare the results of ART treatment in patients with and without endometriosis in a large cohort of patients from different centers over an extented period of time. METHODS: This retrospective study is using data from patients undergoing 27,294 cycles of IVF/ICSI treatment between 1995 and 2011 that were registered in the database of the Latin American Registry maintained by the Latin America Network of Assisted Reproduction. RESULTS: The mean number of retrieved oocytes was higher in the control group, but the mean number of metaphase II oocytes was similar. Fertilization rate and transfer rate were higher in the control group. We observed higher pregnancy rates, per cycle initiated and per embryo transfer and higher live birth rate in the endometriosis group. In the group of patients with 25-35 years old, the number of oocytes, fertilization rate, and number of transferred embryos were significantly higher in the control group. However, pregnancy rate and live birth rate were higher in the endometriosis group. In the group of patients with 36-40 years old, the number of transferred embryos was higher in the control group, but the pregnancy rate and live birth rate were higher in the endometriosis group. In the group of patients with 41 to 42 years old, the number of transferred embryos and the transfer rate were higher in the control group, but the pregnancy rate was higher in the endometriosis group. CONCLUSION: Our results demonstrate that endometriosis does not affect the outcome of patients subjected to IVF/ICSI and although patients with endometriosis present lower number of oocytes and higher cancelation rate, these shortcomings do not reduce pregnancy and live birth rates.

The impact of endometriosis on the outcome of Assisted Reproductive Technology.

BACKGROUND: Endometriosis has been described to impair fertility through various mechanisms. However, studies evaluating the reproductive outcomes of women undergoing assisted reproductive technologies show controversial results. The aim of this study is to assess whether the reproductive outcome is impaired among women with endometriosis-associated infertility undergoing IVF. METHODS: A retrospective cohort study was performed, including women undergoing IVF reported by the Red Latinoamericana de Reproduccion Asistida (Redlara) registry, between January 2010 and December 2012. The study group included women with endometriosis-associated infertility, and the control group women with tubal factor, endocrine disorders or unexplained infertility. Women above 40 years, severe male factor and premature ovarian failure were excluded. The reproductive outcomes of between both groups were compared. The primary outcome was live birth. Secondary outcomes included clinical pregnancy, miscarriage, number of oocytes retrieved and number of fertilized oocytes. Outcomes were assessed after the first fresh IVF cycle, and were adjusted for age and number of embryos transferred. RESULTS: A total of 22.416 women were included (3.583 with endometriosis and 18.833 in the control group). Mean age of patients in the endometriosis group and control group was 34.86 (3.47) and 34.61 (3.91) respectively, p = 0.000. The mean number of oocytes retrieved were 8.89 (6.23) and 9.86 (7.02) respectively, p = 0.000. No significant differences were observed between groups in terms of live birth (odds ratio (OR) 1.032, p = 0.556), clinical pregnancy (OR 1.044, p = 0.428) and miscarriage rates (OR 1.049, p = 0.623). Women with endometriosis had significantly lower number of oocytes retrieved (incidence risk ratio (IRR) 0.917, 95% CI 0.895-0.940), however, the number of fertilized oocytes did not differ among the two groups when adjusting for the number of oocytes retrieved (IRR 1.003, p = 0.794). An age-stratified analysis was performed, and no differences were observed in the reproductive outcomes between groups for women aged under 35 and 35 to 40. CONCLUSIONS: Reproductive outcomes among women undergoing IVF and diagnosed with endometriosis-associated infertility do not differ significantly from women without the disease. Although women with endometriosis generate fewer oocytes, fertilization rate is not impaired and the likelihood of achieving a live birth is also not affected.

Outdoor air pollution and sperm quality.

UNLABELLED: Exposure to air pollution has been clearly associated with a range of adverse health effects, including reproductive toxicity, but its effects on male semen quality are still unclear. We performed a systematic review (up to June 2016) to assess the impact of air pollutants on sperm quality. We included 17 semi-ecological, panel, and cohort studies, assessing outdoor air pollutants, such as PM2.5, PM10, NOx, SO2, and O3, and their effects on DNA fragmentation, sperm count, sperm motility, and sperm morphology. Thirteen studies assessed air pollution exposure measured environmentally, and six used biomarkers of air pollution exposure (two did both). We rated the studies using the Newcastle-Ottawa Scale and assessed with the exposure method. Taking into account these factors and the number of studies finding significant results (positive or negative), the evidence supporting an effect of air pollution on DNA fragmentation is weak but suggestive, on sperm motility is limited and probably inexistent, on lower sperm count is inconclusive, and on sperm morphology is very suggestive. Because of the diversity of air pollutants and sperm parameters, and the studies' designs, we were unable to perform a meta-analysis. In summary, most studies concluded that outdoor air pollution affects at least one of the four semen quality parameters included in the review. However, results lack consistency, and furthermore, studies were not comparable. Studies using standardized air pollution and semen measures are required to obtain more reliable conclusions. PROSPERO REGISTRATION NUMBER: CRD42015007175.

Ratio of progesterone-to-number of follicles as a prognostic tool for in vitro fertilization cycles.

PURPOSE: To establish a ratio of the P level to the number of follicles (P/F ratio) on the day of human chorionic gonadotropin (hCG) administration and to evaluate whether this ratio is associated with in vitro fertilization (IVF) outcome. METHODS: This study was conducted between January 2012 and June 2013. A total of 337 patients with cleavage-stage day-3 fresh embryo transfer with P levels ≤1.5 ng/mL on the day of hCG administration were included in the study. The main outcome was ongoing pregnancy rate. RESULT(S): The P/F ratio was calculated according to the equation (P[ng/mL]/number of follicles) on the day of final oocyte maturation. Using ROC, we established a cut-off level of 0.075 for the P/F ratio. The area under the curve (AUC) (0.756; 95 % confidence interval [CI]: 0.704-0.807) indicated that it was a good prognostic test. In group 1 (patients under 36 years old), the ongoing pregnancy rates were 57 and 30 % for patients with P/F ratios ≤ .075 and > .075, respectively, (p = 0.003). In group 2 (patients between 36 and 39 years old), the ongoing pregnancy rates were 58 % and 17 % (p = 0.001) for patients with P/F ratios ≤ .075 and > .075, respectively. In group 3 (patients ≥ 40 years old), the ongoing pregnancy rates were 41.7 and 10.9 % (p = 0.001) for patients with P/F ratios ≤ .075 and > .075, respectively. CONCLUSIONS: The P/F ratio is a good prognostic test for predicting IVF outcome that can correlate the P level with ovarian response.

The impact of donor insemination on the risk of preeclampsia: a systematic review and meta-analysis.

A systematic review and meta-analysis were performed to evaluate whether women who conceive with donor sperm have an increased risk of preeclampsia compared with those who use their partner's sperm. Studies that compared women who were impregnated by donor and partner sperm were included. The main outcomes assessed were preeclampsia and gestational hypertension rates. Altogether, 10,898 women (2342 pregnancies by donor sperm versus 8556 by the partner's sperm) were included from seven observational studies. Conception using donor sperm was associated with an increased risk of preeclampsia (odds ratio [OR] 1.63, 95% CI 1.36-1.95) compared with using a partner's sperm. No difference was observed in the risk of gestational hypertension (OR 0.94, 95% CI 0.43-2.03). In conclusion, pregnancies achieved by donor sperm significantly increase the risk of preeclampsia, although the underlying mechanisms remain unclear. Additional studies are required to confirm these findings.

Alcohol consumption and in vitro fertilization: a review of the literature.

The aim of our study is to determine whether alcohol consumption affects the results of in vitro fertilization. A review of the literature was performed to find prospective cohort studies of couples undergoing in vitro fertilization in which alcohol intake was recorded. A primary search returned 389 studies, 2 of which were finally considered eligible. A total of 2908 couples were analyzed in terms of pregnancy outcomes depending on drinking habits. The risk of IVF failure increased 4.14-fold and 2.86-fold with an increased alcohol intake of 12 gr/d in women during the week and month before, respectively. The odds ratio (OR) of live birth rate in women who drank at least four drinks per week compared with women who drank fewer was 0.84; this difference was statistically significant. Paternal alcohol use levels 1 month, 1 week and during the attempts were also associated with worse reproductive effects. Our review, though including a small number of studies that were heterogeneous in design, revealed decreased rates of pregnancy and fertilization outcomes for couples who drank before or during their in vitro fertilization techniques. This suggests that couples undergoing IVF should be advised to abstain from alcohol prior to and during their procedures.

High-grade squamous intraepithelial lesion could be managed conservatively in women up to 25 years: results from a retrospective cohort study.

OBJECTIVE: The aim of this study was to compare histologic findings and clinical outcomes of women up to 25 years with a high-grade squamous intraepithelial lesion (HSIL) compared to women older than 25 years. MATERIALS AND METHODS: Sixty-three women up to 25 years and 245 women older than 25 years with HSIL, diagnosed from June 1991 to September 2008, were examined and treated following the official Spanish guidelines. Colposcopic and histologic findings and needs for treatment were recorded, and patients were followed up for a minimum of 12 months. RESULTS: A total of 308 patients were evaluated; 63.49% of women up to 25 years and 77.10% of women older than 25 years with HSIL had cervical intraepithelial neoplasia (CIN) 2+ histology (p = .04). Also, 74.60% of women up to 25 years and 99.24% of women older than 25 years underwent an excisional procedure (p < .001). No significant or clinical differences were found in the 1-year follow-up outcomes (82.54% vs 78.37% had normal results; p = ns). CONCLUSIONS: Women up to 25 years have less CIN 2+ histologic findings and less need for conization compared to older women. Our findings support the feasibility to design an adequate protocol for these younger women, which would be less aggressive and would, consequently, minimize obstetric long-term secondary effects.

Changes in DNA fragmentation during sperm preparation for intracytoplasmic sperm injection over time.

OBJECTIVE: To compare the DNA fragmentation of semen samples established by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL) after incubation in polyvinylpyrrolidone (PVP) and hyaluronic acid (HA) for different time periods. DESIGN: Comparative prospective study. SETTING: Center for reproductive medicine. PATIENT(S): Twenty-seven semen samples from infertile patients. INTERVENTION(S): None. METHODS: Semen analysis and DNA fragmentation assays (TUNEL) were performed. Two groups were established: A) normal TUNEL (<20%); and B) Abnormal TUNEL (≥ 20%). TUNEL was performed in neat (T0), postgradient (TG), 1-hour postgradient (TG1), and 2-hour postgradient (TG2) samples and in TG2 samples after 0.5, 1.0, and 1.5 hours of incubation in PVP or HA. RESULT(S): TUNEL levels were significantly reduced after gradient separation compared with neat values. In group A, TUNEL levels were significantly higher in the TG2 + 1.5 hours in PVP and HA samples but did not reach abnormal levels. In group B, TUNEL levels were significantly higher in the TG2 + 1 hour in PVP and HA samples. CONCLUSION(S): Sperm DNA fragmentation significantly decreased after centrifugation gradient, regardless of the initial levels of the sample. Samples with TUNEL ≥ 20% were more susceptible to a significant increase in DNA fragmentation over time, with similar increases being observed over time for samples that were incubated in HA or PVP. These data may be relevant for sperm preparation for intracytoplasmic sperm injection.

Sperm selection using magnetic activated cell sorting (MACS) in assisted reproduction: a systematic review and meta-analysis.

PURPOSE: To determine whether the use of Magnetic Activated Cell Sorting (MACS) as a sperm selection technique improves ART success rates in couples undergoing assisted reproduction treatment. METHODS: Systematic review and meta-analysis of prospective randomized trials. Two reviewers conducted study selection and data extraction independently. RESULTS: Five studies (prospective randomized trials) that comprised 499 patients were included. Sperm selection using MACS resulted in statistically significant differences in pregnancy rates when compared with density gradient centrifugation and swim-up techniques (RR=1.50, 95 % CI 1.14-1.98). No differences were found between the groups according to the implantation (RR=1.03, 95 % CI 0.80-1.31) and miscarriage (RR=2.00, 95 % CI 0.19-20.90) rates. CONCLUSIONS: MACS appears to be a safe and efficient method to select functional sperm with consistently good results. This technique may improve pregnancy rates when used to complement standard sperm selection methods in ART.

Fresh embryo transfer versus frozen embryo transfer in in vitro fertilization cycles: a systematic review and meta-analysis.

OBJECTIVE: To examine the available evidence to assess if cryopreservation of all embryos and subsequent frozen embryo transfer (FET) results in better outcomes compared with fresh transfer. DESIGN: Systematic review and meta-analysis. SETTING: Centers for reproductive care. PATIENT(S): Infertility patient(s). INTERVENTION(S): An exhaustive electronic literature search in MEDLINE, EMBASE, and the Cochrane Library was performed through December 2011. We included randomized clinical trials comparing outcomes of IVF cycles between fresh and frozen embryo transfers. MAIN OUTCOME MEASURE(S): The outcomes of interest were ongoing pregnancy rate, clinical pregnancy rate, and miscarriage. RESULT(S): We included three trials accounting for 633 cycles in women aged 27-33 years. Data analysis showed that FET resulted in significantly higher ongoing pregnancy rates and clinical pregnancy rates. CONCLUSION(S): Our results suggest that there is evidence that IVF outcomes may be improved by performing FET compared with fresh embryo transfer. This could be explained by a better embryo-endometrium synchrony achieved with endometrium preparation cycles.

Paraoxonase-1 is only present in traceable amounts in seminal fluid and does not show any relationship with male subfertility.

UNLABELLED: Study Type - Aetiology (case series). LEVEL OF EVIDENCE: 4. What's known on the subject? and What does the study add? Oxidative stress seems to be one of the biochemical causes of defective sperm function. Paraoxonases are antioxidant enzymes that degrade lipid peroxides. There is a paucity of data on the possible role played by these enzymes in the pathophysiology of male sub-fertility. The present study shows that testicular tissue of sub-fertile patients clearly expresses paraoxonases-1, 2, and 3. These findings suggest a role for these enzymes in the protection against lipid peroxidation inside the cell. However, the concentration and activity of paraoxonase-1 in semen are negligible and are probably the result of cellular catabolism, with no significant biological function. OBJECTIVE: To characterise the immunohistochemical sites of paraoxonase (PON) 1, PON2 and PON3 in human testicular tissue, and to analyse PON1 levels in semen, aiming to investigate the role played by these enzymes in the pathophysiology of male subfertility. PATIENTS AND METHODS: The present study was performed in 41 semen samples from normal donors and in 52 semen samples and ten testicle biopsies from patients who were being evaluated for causes of subfertility. RESULTS: Immunohistochemical analyses showed high levels of PON1 and PON3 expression in testicular tissue. PON2 expression was also detected, albeit at weaker levels. Oxidative stress indicators in biopsies were low and localized in some specific areas of the seminiferous tubules. PON1 was detected in seminal fluid at very low levels but with no significant differences between patients and controls. Receiver-operating characteristic analysis showed a low diagnostic power of semen PON1 levels. CONCLUSIONS: The present study shows high protein expression levels of PON1, PON2 and PON3 in testicular cells. The concentrations and activities of PON1 in semen are negligible and are probably the result of cellular catabolism, with no significant biological function in the testes.