Assessing the impact of delayed blastulation using time lapse morphokinetics and preimplantation genetic testing in an IVF patient population.

PURPOSE: There is clinical evidence that early cleavage timing parameters predictive of blastocyst development also correlate with embryo implantation potential. The aim of this study is to determine the developmental competency of embryos with delayed blastulation. METHODS: Retrospective study performed from 2015 to 2016 at the Division of Reproductive Endocrinology and Infertility at Northwestern University. RESULTS: A total of 2,292 embryos from 524 patients were included. Day 6 blastocysts had statistically significant longer times for every time point analyzed than day 5 blastocysts (p < 0.001). We found no statistically significant difference in euploidy rates between day 5 (44%) and day 6 (41%) embryos (p = 0.573). t7 and t8 time points were independent predictors of euploidy after controlling for day of biopsy (p < 0.015 and p < 0.014, respectively). Intrauterine pregnancy (IUP) and live birth (LB) were less likely to occur after transferring day 6 embryos (p = 0.0033 and p = 0.0359) without previous genetic testing. However, in embryos that undergo preimplantation genetic testing for aneuploidy (PGT-A), there were no significant differences in IUP or LB rates. CONCLUSION: Early time-lapse points can be used to predict embryo development. Day of blastulation may be an independent predictor IUP, with day 6 blastocysts having lower pregnancy and live birth rates. Our data suggests that day 5 and day 6 PGT-A tested embryos show similar rates of euploidy, suggesting that differences in PR seen in the non-PGT-A tested group may be caused by factors other than aneuploidy. Genetic testing technologies in combination with time-lapse microscopy may provide further information to improve IVF outcomes.

Do patients who achieve pregnancy using IVF-PGS do the recommended genetic diagnostic testing in pregnancy?

PURPOSE: Patients undergoing in-vitro fertilization (IVF) with preimplantation genetic screening (PGS) are counseled about the limitations of this technique. As part of the consent process for PGS, physicians recommend diagnostic genetic testing performed in early pregnancy to definitively rule out chromosomal abnormalities. We have noted anecdotally, however, that few patients undergo the recommended diagnostic testing. In this study, we are examining if women who conceived using IVF-PGS did early pregnancy chromosomal testing, and if they did, what type of testing they had. METHODS: This study was performed from 2015 to 2017 in the Division of Reproductive Endocrinology and Infertility at Northwestern University. We included patients who became pregnant after IVF-PGS who were seen by the Division of Reproductive Genetics and non-PGS control group. RESULTS: Sixty-eight patients were included. A total of 50 patients (73.5%) opted for non-invasive prenatal screening; 5 (7.4%) had invasive testing (4 had chorionic villus sampling and 1 had amniocentesis). A total of 13 patients (19%) declined further genetic testing. When comparing demographic data, the mean age was significantly higher in the group of patients who pursued non-invasive testing than in the group who declined further testing (37.15 vs 34.05 years old, p < 0.05). Control group declined invasive diagnostic testing. CONCLUSIONS: Most patients who conceive using IVF-PGS do not pursue diagnostic prenatal chromosomal testing. Future studies focusing on decision making in this patient group are warranted to further elucidate why a small percentage of patients opt for diagnostic testing, even when adequately counseled about the inherent limitations of PGS.

Reproductive Outcomes After Hysteroscopic Resection of Retained Products of Conception.

STUDY OBJECTIVE: To characterize pregnancy outcomes after hysteroscopic resection of retained products of conception (RPOCs), especially as it relates to abnormal placentation. DESIGN: Retrospective cohort study (Canadian Task Force classification II-2). SETTING: Academic medical center. PATIENTS: All women who underwent hysteroscopic resection of RPOCs at Northwestern Prentice Women's Hospital between January 2004 and December 2014. INTERVENTIONS: Hysteroscopic resection of RPOCs. MEASUREMENTS AND MAIN RESULTS: The medical records of all cases of hysteroscopic resection of RPOCs between January 2004 and December 2014 were reviewed. Demographic characteristics, operative findings, surgical procedure, surgical pathology, and pregnancy outcomes for preceding and subsequent pregnancies were obtained. Our primary outcome was abnormal placentation in the pregnancy after the procedure. There were a total of 55 subsequent pregnancies and 38 live births. Among these pregnancies, 54.5% (30/55) were vaginal deliveries, 34.5% (19/55) were cesarean deliveries, and 7.3% (4/55) were early pregnancy losses. Abnormal placentation was present in 18.1% of subsequent pregnancies (10/55). This consisted of 3 patients with placenta previa, 2 with placenta accreta, and 5 with retained placenta. CONCLUSION: Women who undergo hysteroscopic resection of RPOCs have a higher rate of abnormal placentation in subsequent pregnancies when compared with the general population. Although the etiology is likely multifactorial, the underlying pathology leading to the initial diagnosis of RPOCs is believed to play a major role.

First contact: the intersection of demographics, knowledge, and appraisal of treatment at the initial infertility visit.

OBJECTIVE: To determine the impact of the initial infertility visit on treatment-related knowledge, patient anxiety, and appraisals of treatment. DESIGN: Prospective survey. SETTING: Academic medical center. PATIENT(S): Two hundred thirty-four English-speaking women aged 18-50 years attending their first infertility visit. INTERVENTION(S): Participants completed a survey assessing health literacy, knowledge, anxiety, and appraisals of the treatment process before and after their infertility visit. MAIN OUTCOME MEASURE(S): Knowledge of infertility and treatment and anxiety and appraisal scores. RESULT(S): Most participants were white and earned >$100,000/year and had at least a college education. Baseline knowledge of reproductive anatomy, assisted reproductive technology (ART), and fertility factors was modest but improved after the initial visit. Factors associated with higher knowledge included higher education and income, white or Asian ethnicity, and English as a primary language. Patient appraisals of treatment represented by the positive (Challenge) and negative (Threat and Loss) subscale scores on the Appraisal of Life Events (ALE) scale changed over time Negative appraisals of treatment and anxiety scores decreased and positive appraisals of treatment increased after the initial visit. Lower knowledge was associated with higher positive appraisal scores; lower health literacy was associated with higher anxiety and appraisal scores (positive and negative) after the visit. Black women had higher Challenge scores compared with white and Asian women. Hispanic women had higher anxiety scores than non-Hispanic women. CONCLUSION(S): Infertility patients have modest baseline knowledge of fertility and infertility treatment. The initial infertility visit can improve this knowledge and decrease both negative appraisals of treatment and anxiety levels. Differences in knowledge and appraisal were seen across ethnic groups and other demographic variables. Physicians should individualize patient counseling to improve patients' knowledge and provide realistic treatment expectations while also reducing patient anxiety.

Ureteral Trauma During Transvaginal Ultrasound-Guided Oocyte Retrieval: A Case Report.

BACKGROUND: Transvaginal ultrasound-guided oocyte retrieval is a safe and well-tolerated procedure. Complications are uncommon and usually limited to vaginal bleeding and pelvic infection. Ureteral injury following oocyte retrieval is exceedingly rare, with only 8 previously reported cases. CASE: A 34-year-old woman developed gross hematuria 4 hours after transvaginal ultrasound-guided oocyte aspiration. Cystoscopy, laparoscopy, and retrograde pyelography revealed bleeding from the left ureter, no intra-abdominal bleeding, and a patent left urinary collecting system. The ureteral bleeding was successfully managed with placement of a ureteral stent. CONCLUSION: Ureteral trauma during transvaginal-guided oocyte retrieval is a rare complication with a variable clinical presentation. If ureteral injuries are not promptly recognized, significant morbidity may occur. This case demonstrates that early identification of injury and timely intervention result in favorable outcomes.

Randomized controlled trial: effects of acupuncture on pregnancy rates in women undergoing in vitro fertilization.

OBJECTIVE: To evaluate the influence of "true" versus "sham" acupuncture on pregnancy rates (PRs) in women undergoing IVF. DESIGN: Randomized controlled trial, double-blinded with independent observer. SETTING: Academic infertility clinic. PATIENT(S): One hundred sixty patients <38 years old undergoing IVF with or without intracytoplasmic sperm injection. INTERVENTION(S): Subjects were randomly allocated to the true or sham group and underwent acupuncture 25 minutes before and after ET. Subjects completed a McGill Pain Questionnaire regarding their clinical symptoms during ET. MAIN OUTCOME MEASURE(S): Clinical PR and clinical symptoms during ET. RESULT(S): While the overall clinical PR was 51.25%, there was no significant difference between the arms of the study (true = 45.3% vs. sham = 52.7%); 33.1% of the patients had ultrasound-documented singleton pregnancy, and 15% of patients had twin gestations, while one patient in the true arm had a triplet gestation. There were significant differences in the subjective, affective, and total pain experience between both arms. The subjects in the true arm described their acupuncture session as being more "tiring" and "fearful" and experienced more "achiness" compared with their sham counterparts. CONCLUSION(S): There was no statistically significant difference in the clinical or chemical PRs between both groups. Patients undergoing true acupuncture had differing sensory experiences compared with patients in the sham arm. There were no significant adverse effects observed during the study, suggesting that acupuncture is safe for women undergoing ET.

Can Doppler ultrasound-guided oocyte retrieval improve IVF safety?

Transvaginal ultrasound-guided oocyte retrieval has gained universal acceptance with an excellent safety record overall. However, even with contemporary ultrasound resolution, the aspiration needle can injure adjacent pelvic organs and blood vessels and result in external and internal bleeding. Although the idea that Doppler ultrasound might reduce the risk of blood vessel injury during follicular aspiration seems to be plausable, measurement of peritoneal blood loss and the validity of this opinion has never been appropriately tested. Using a proposed classification method in an IVF programme, it was estimated that a significant peritoneal bleeding occurred in 56/898 (6%) of IVF patients. Although Doppler ultrasound was routinely used in all patients, it did not predict 24/53 (45%) of the patients with moderate peritoneal bleeding. In 8/53 cases (15%) with moderate peritoneal bleeding, vaginal bleeding was also detected and correctly predicted during oocyte aspiration using colour Doppler vaginal vessel imaging. Colour Doppler ultrasound guidance is an easily accessible technology with a theoretical promise to improve IVF safety and, with proper usage, has the potential to reduce haemorrhagic complications.

Steroidogenic factor-1 and endometriosis.

Endometriosis is a common and chronic disease characterized by persistent pelvic pain and infertility. Estradiol is essential for growth and inflammation in endometriotic tissue. The complete cascade of steroidogenic proteins/enzymes including aromatase is present in endometriosis leading to de novo estradiol synthesis. PGE(2) induces the expression of the genes that encode these enzymes. Upon PGE(2) treatment, coordinate recruitment of the nuclear receptor SF-1 to the promoters of these steroidogenic genes is the key event for estradiol synthesis. SF-1 is the key factor determining that an endometriotic cell will respond to PGE(2) by increased estradiol formation. The presence of SF-1 in endometriosis and its absence in endometrium is determined primarily by the methylation of its promoter. The key steroidogenic enzyme in endometriosis is aromatase encoded by a single gene because its inhibition blocks all estradiol biosynthesis. Aromatase inhibitors diminish endometriotic implants and associated pain refractory to existing treatments in affected women.

La migración de la burbuja de aire postransferencia embrionaria es un evento al azar / Air bubble migration post embrionary tranferir is a random event

Demostrar que la migración de la burbuja de aire postransferencia embrionaria es un evento al azar que no depende de la posición del paciente. Estudio prospectivo multicéntrico. Se practicó transferencia embrionaria a 69 pacientes bajo guía ultrasonográfica. Se cargó el catéter de transferencia con una o dos burbujas junto con el medio conteniendo los embriones usando catéter de Frydman o catéter de Wallace y luego se les hizo seguimiento ecográfico en posición horizontal, 20-25 minutos más tarde y luego a los 90 minutos postransferencia en posición vertical para verificar movimiento y configuración de las burbujas. Centro Médico Docente La Trinidad, Caracas. Nortwestern University Feimberg School of medicine, Chicago, IL. EE.UU. En 50 (72,46 por ciento) de las pacientes se observó movimiento de las burbujas. En posición de pie se demostró una distribución al azar sin efecto visible de la gravedad. Las burbujas de aire se mueven y dividen frecuentemente posterior a la transferencia con la paciente en posición horizontal y de pie, lo que sugiere actividad contráctil uterina. No fue común observar movimiento de burbujas relacionada con la gravedad, lo que sugiere que el descanso horizontal postransferencia embrionaria puede ser innecesario.

To demonstrate that air bubble migration is a random event after embryo transfer regardless of the position of patient. Multicenter prospective study. Sixty nine embryo transfers were performed under ultrasound guidance. Transfer catheter was loaded with one or two air bubbles and medium containing embryos, either Wallace catheter or Frydman catheter were used, ultrasound tracking of air bubble was performed to verify movement and configuration inmediately, 20-25 minutes in horizontal position and 90 minutes on standing position after embryo transfer. Centro Medico Docente La Trinidad, Caracas. Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA. Movement was observed in 50 (72,46 percent) of the patients. Movement was not observed related to gravity while standing up. Air bubbles move and split frequently after embryo transfer with the patient in the horizontal position and on standing, suggestive of active uterine contractions. Gravity related bubble motion was uncommon, suggesting that horizontal rest post embryo transfer may be unnecessary.

Promoter methylation regulates estrogen receptor 2 in human endometrium and endometriosis.

Steroid receptors in the stromal cells of endometrium and its disease counterpart tissue endometriosis play critical physiologic roles. We found that mRNA and protein levels of estrogen receptor 2 (ESR2) were strikingly higher, whereas levels of estrogen receptor 1 (ESR1), total progesterone receptor (PGR), and progesterone receptor B (PGR B) were significantly lower in endometriotic versus endometrial stromal cells. Because ESR2 displayed the most striking levels of differential expression between endometriotic and endometrial cells, and the mechanisms for this difference are unknown, we tested the hypothesis that alteration in DNA methylation is a mechanism responsible for severely increased ESR2 mRNA levels in endometriotic cells. We identified a CpG island occupying the promoter region (-197/+359) of the ESR2 gene. Bisulfite sequencing of this region showed significantly higher methylation in primary endometrial cells (n = 8 subjects) versus endometriotic cells (n = 8 subjects). The demethylating agent 5-aza-2'-deoxycytidine significantly increased ESR2 mRNA levels in endometrial cells. Mechanistically, we employed serial deletion mutants of the ESR2 promoter fused to the luciferase reporter gene and transiently transfected into both endometriotic and endometrial cells. We demonstrated that the critical region (-197/+372) that confers promoter activity also bears the CpG island, and the activity of the ESR2 promoter was strongly inactivated by in vitro methylation. Taken together, methylation of a CpG island at the ESR2 promoter region is a primary mechanism responsible for differential expression of ESR2 in endometriosis and endometrium. These findings may be applied to a number of areas ranging from diagnosis to the treatment of endometriosis.

Transcriptional activation of steroidogenic factor-1 by hypomethylation of the 5' CpG island in endometriosis.

CONTEXT: Endometriosis is an estrogen-dependent disease. Steroidogenic factor-1 (SF-1), a transcriptional factor essential for activation of multiple steroidogenic genes for estrogen biosynthesis, is undetectable in normal endometrial stromal cells and aberrantly expressed in endometriotic stromal cells. OBJECTIVE: The objective of the study was to unravel the mechanism for differential SF-1 expression in endometrial and endometriotic stromal cells. DESIGN: We identified a CpG island flanking the SF-1 promoter and exon I region and determined its methylation patterns in endometrial and endometriotic cells. SETTING: The study was conducted at Northwestern University. PATIENTS OR OTHER PARTICIPANTS: Eutopic endometrium from disease-free subjects (n = 8) and the walls of cystic endometriosis lesions of the ovaries (n = 8) were investigated. INTERVENTION(S): Stromal cells were isolated from these two types of tissues. MAIN OUTCOME MEASURE(S): Measures are mentioned in Results. RESULTS: SF-1 mRNA and protein levels in endometriotic stromal cells were significantly higher than those in endometrial stromal cells (P < 0.001). Bisulfite sequencing showed strikingly increased methylation in endometrial cells, compared with endometriotic cells (P < 0.001). Demethylation by 5-aza-2'-deoxycytidine increased SF-1 mRNA levels by up to 55.48-fold in endometrial cell (P < 0.05). Luciferase assays showed that the -85/+239 region bearing the CpG island regulated its activity (P < 0.01). Natural or in vitro methylation of this region strikingly reduced SF-1 promoter activity in both cell types (P < 0.01). Chromatin immunoprecipitation assay showed that methyl-CpG-binding domain protein 2 binds to the SF-1 promoter in endometrial but not endometriotic cells. CONCLUSIONS: This is the first demonstration of methylation-dependent regulation of SF-1 in any mammalian tissue. These findings point to a new mechanism for targeting local estrogen biosynthesis in endometriosis.

Abnormal uterine bleeding, a new terminology is needed.

The current terminology describing abnormal uterine bleeding (AUB) is not universally agreed upon. A new AUB terminology must be uniform, unambiguous, and symptom focused. A prototype letters and numbers formula covering the entire spectrum of AUB is proposed that may contain sequences such as B1D2F1A2P1 (i.e., moderate-1 bleeding, lasting for 2 days-duration, with frequent monthly-1 presentation, causing significant-anemia and mild-1 pains). Universal acceptance of a similar standard and reproducible terminology will add semiquantitative information, enable accurate reporting, and facilitate clinical research.

Studies of gene expression in human cumulus cells indicate pentraxin 3 as a possible marker for oocyte quality.

OBJECTIVE: To search for differentially expressed genes in cumulus cells from two groups of oocytes with different developmental outcome in vitro. DESIGN: Analyses of gene expression in human cumulus cells from oocytes that failed to fertilize in vitro (group A) and those that developed into normal-appearing embryos on day 3 (group B). SETTING: University-based facilities for clinical service and research. PATIENT(S): Women undergoing IVF treatment for infertility. INTERVENTION(S): Cumulus cells were collected from oocytes that were aspirated from ovarian follicles for IVF. The oocytes were cultured individually for IVF and embryo development. Total RNA was extracted from the cumulus cells for gene expression analyses. MAIN OUTCOME MEASURE(S): General gene expression profiles and relative abundance of pentraxin 3 (Ptx3) mRNA. RESULT(S): DNA microarray analysis identified 160 genes, including Ptx3, that were differentially expressed between cumulus cells in group A and group B. Quantitative analysis confirmed that the relative abundance of Ptx3 mRNA in cumulus cells was highly associated with oocyte development. CONCLUSION(S): This study demonstrated that changes in the expression levels of 160 genes, including particularly Ptx3, in human cumulus cells may be indicative of the quality of their enclosed oocyte.

Increased endometrial thickness is associated with improved treatment outcome for selected patients undergoing in vitro fertilization-embryo transfer.

OBJECTIVE: To examine possible relationships between endometrial thickness and treatment outcome after IVF and embryo transfer, and to explore the role of potential confounding factors that may influence such relationships. DESIGN: Retrospective study. SETTING: A university-affiliated clinical IVF center. PATIENT(S): Patients undergoing IVF-embryo transfer with their own oocytes. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Endometrial thickness was determined on the day of hCG administration, 2 days before oocyte retrieval. Clinical pregnancy was confirmed by ultrasound observation of fetal heart activity. RESULT(S): The study analyzed 897 IVF-embryo transfer cycles. Treatment outcome (clinical pregnancy) after IVF-embryo transfer was positively associated with increased endometrial thickness and peak E(2) concentrations in serum, and negatively associated with advanced age. Endometrial thickness was dependent on peak E(2) concentrations in serum, but was independent of patient age or duration of ovarian stimulation. Thin endometrium reduced PRs in relatively young patients (<38 years old), in patients who required more than 10 days of gonadotropin stimulation, or in patients whose embryo transfers consisted of poor quality embryos. CONCLUSION(S): Increased endometrial thickness was associated with improved treatment outcome, but this association was dependent on patient age, duration of ovarian stimulation, and embryo quality.

Electrodessication of a cervical heterotopic pregnancy.

A cervical heterotopic assisted reproductive technology (ART) pregnancy was diagnosed sonographically. The cervical pregnancy was electrodesiccated with a Kleppinger device. The intrauterine pregnancy was carried to term.

Delay of embryo transfer to day 5 results in decreased initial serum beta-human chorionic gonadotropin levels.

OBJECTIVE: To investigate the impact of delaying the transfer of in vitro fertilized embryos to day 5 on embryo development by comparing serum beta-hCG levels in pregnancies established after day 3 embryo transfers and those after day 5 blastocyst transfers at equivalent time intervals after fertilization. DESIGN: A retrospective analysis. SETTING: Assisted reproductive technology center in an academic research environment. PATIENT(S): Women who had an embryo transfer procedure performed after in vitro fertilization (IVF) at Northwestern University's IVF Program between January 1999 and December 2001 were included in this study. INTERVENTION(S): Embryo transfer was performed either on day 3 or day 5 after oocyte retrieval for IVF (day 0), depending on the availability of good-quality embryos on day 3. MAIN OUTCOME MEASURE(S): Serum beta-hCG concentrations determined 13 and 15 days after fertilization in pregnancies established by transferring cleavage-stage embryos on day 3 or blastocysts on day 5. RESULT(S): In singleton pregnancies, serum beta-hCG concentrations were 75 +/- 54 (mean +/- SD, n = 203) or 62 +/- 41 (n = 109) IU/mL after day 3 or day 5 transfers, respectively. In twin pregnancies, the beta-hCG concentrations were 162 +/- 105 (n = 52) or 109 +/- 55 (n = 49) after day 3 or day 5 transfers, respectively. The percentage increases in beta-hCG concentrations between the first and second measurements were similar in the two groups (day 3: 144 +/- 109, day 5: 142 +/- 63, not statistically significant). CONCLUSION(S): Initial beta-hCG concentrations in pregnancies resulting from day 5 transfers were lower than those from day 3 transfers when assessed at equivalent intervals from fertilization. This suggests that embryo development or implantation may be impaired by the additional 2 days in culture.

Relationship between peak serum estradiol levels and treatment outcome in in vitro fertilization cycles after embryo transfer on day 3 or day 5.

OBJECTIVE: To examine the relationships between peak serum estradiol (E(2)) levels and treatment outcome in in vitro fertilization (IVF) cycles after embryo transfer (ET) on day 3 or day 5. DESIGN: Retrospective analysis of 697 IVF-ET cycles between January 1999 and December 2001. SETTING: A university-affiliated assisted reproduction program. PATIENT(S): Infertile patients undergoing IVF-ET cycles. INTERVENTION(S): Peak E(2) concentration in serum was determined on the day of human chorionic gonadotropin (hCG) administration. The IVF-generated embryos were cultured for 2 days until transfer on day 3. If more than four 8-cell embryos were present on day 3, embryo culture was continued until day 5 for blastocyst transfer. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates. RESULT(S): High peak E(2) levels did not adversely affect treatment outcome. After the cycles were divided according to the day of ET, high peak E(2) levels were associated with improved pregnancy rates after ET on day 5 but not on day 3. CONCLUSION(S): Increasing peak E(2) levels in IVF cycles are associated with improved pregnancy rates after ET on day 5.