Human embryo live imaging reveals nuclear DNA shedding during blastocyst expansion and biopsy.

Proper preimplantation development is essential to assemble a blastocyst capable of implantation. Live imaging has uncovered major events driving early development in mouse embryos; yet, studies in humans have been limited by restrictions on genetic manipulation and lack of imaging approaches. We have overcome this barrier by combining fluorescent dyes with live imaging to reveal the dynamics of chromosome segregation, compaction, polarization, blastocyst formation, and hatching in the human embryo. We also show that blastocyst expansion mechanically constrains trophectoderm cells, causing nuclear budding and DNA shedding into the cytoplasm. Furthermore, cells with lower perinuclear keratin levels are more prone to undergo DNA loss. Moreover, applying trophectoderm biopsy, a mechanical procedure performed clinically for genetic testing, increases DNA shedding. Thus, our work reveals distinct processes underlying human development compared with mouse and suggests that aneuploidies in human embryos may not only originate from chromosome segregation errors during mitosis but also from nuclear DNA shedding.

Does the birth plan match what is relevant to women? Preferences of Spanish women when giving birth.

BACKGROUND: To support women in making shared decisions, it is important to know what is relevant to them. The aim is to explore which of the options included in birth plans (BP) are of most interest to women, and which are more controversial. In addition, the possible association of this variability with personal characteristics. METHODS: The data are part of a cross-sectional descriptive study, carried out in xxx, on the clinimetric characteristics of two instruments to measure women's needs in labour and postpartum. Women were recruited consecutively by their midwives during pregnancy check-ups, receive a link to a digital questionnaire and were allowed to provide links to the questionnaires to other pregnant women. Women were asked to determine their level of agreement with statements about the birth environment, accompaniment, pain relief, medical intervention and neonatal care. The relationship between agreement with each statement, socio-demographic variables and fear of childbirth (W-DEQ-A) was analysed using a combination of descriptive statistics to analyse frequencies, and regression models to test the effect of socio-demographic variables and fear of childbirth on those items with the greatest variability. RESULTS: Two hundred forty-seven women responded. More than 90% preferred a hospital delivery, with information about and control over medical intervention, accompanied by their partner and continuous skin-to-skin contact with the newborn. There are other questions to which women attach less importance or which show greater variability, related to more clinical aspects, like foetal monitoring, placenta delivery, or cord clamping… Various factors are related to this variability; parity, nationality, educational level, risk factor or fear of childbirth are the most important. CONCLUSIONS: Some items referring to the need for information and participation are practically unanimous among women, while other items on technical interventions generate greater variability. That should make us think about which ones require a decision after information and which ones should be included directly. The choice of more interventional deliveries is strongly associated with fear of childbirth.

Development and evaluation of the psychometric properties of a digital questionnaire for the self-management of health and well-being in the postpartum period.

BACKGROUND: Despite the fact that the Global Strategy for Women's, Children's and Adolescents' Health (2016-2030) recognises the special importance of care for women during the postpartum period, thus highlighting the need to identify and measure any condition that may affect the welfare of pregnant women in any way, this is one of the most neglected stages in the health system. Given the absence in our area of global, efficient instruments, the objective of this study was to design a complete, specific measurement tool with good metric qualities in digital format for the evaluation of self-reported health and well-being during the puerperium, to conform to what was proposed by the ICHOM. METHODS: A cross-sectional study was carried out to evaluate the psychometric characteristics of a digital measurement tool. The development of the tool was carried out in 4 steps, following the recommendations of the International Test Commission. It was tested on 280 puerperas attending primary healthcare appointments in the Basque Healthcare System (Osakidetza), and they did the newly created survey, answering all the questions that had been selected as the gold standard. The average age of the women was 34.93 (SD = 4.80). The analysis of the psychometric characteristics was based on mixed procedures of expert judgment (a focus group of healthcare professionals, an item evaluation questionnaire and interviews with users) and quantitative evaluations (EFA, CFA, and correlation with gold standard, ordinal alpha and McDonald's omega). RESULTS: The final version of the tool comprised 99 items that evaluate functional state, incontinence, sexuality, breastfeeding, adaptation to the role of mother and mental health, and all of these questions can be used globally or partially. It was found that the scores were valid and reliable, which gives metric guarantees for using the tool in our area. CONCLUSIONS: The use of this comprehensive concise tool with good psychometric properties will allow women to take stock of their situation, assess if they have the necessary resources, in psychological and social terms, and work together with midwives and other healthcare professionals on the most deficient areas.

Development and evaluation of the psychometric properties of a digital questionnaire for the evaluation of perinatal psychosocial needs.

BACKGROUND: If the purpose of maternal education is for women to take control of their own health and that of their family in the process, it is essential to have a simple instrument that allows them to self-assess, globally, how prepared they are to face future childbirth and maternity. As there is nothing similar in our area, the objective of this study was to design a complete, specific measurement questionnaire, with good metric quality and in digital format, for the assessment of perinatal psychosocial needs. METHODS: A cross-sectional study was carried out, to evaluate the psychometric properties of a digital measurement questionnaire. The questionnaire was developed in 4 steps following the recommendations of the International Test Commission. The participants were 263 pregnant women who were recruited in primary health care appointments in the Basque Healthcare Service (Osakidetza); they completed the newly created questionnaire and all the test selected as gold standard. Their mean age was 33.55 (SD = 4.73). The analysis of the psychometric characteristics was based on mixed expert judgment procedures (focus group of healthcare professionals, item assessment questionnaire and interviews with users) and quantitative procedures (EFA, CFA, association with the gold standard and classification agreement index, ordinal alpha and McDonald's omega). RESULTS: The final version of the questionnaire was made up of 55 items that evaluate 8 aspects related to perinatal psychosocial well-being (anxious-depressive symptoms, pregnancy acceptance, partner support, coping, internal locus of control, childbirth self-efficacy, perception of childbirth as a medicalized event, and fear of childbirth). Various tests were made of the validity and reliability of the scores, providing metric guarantees for their use in our context. CONCLUSIONS: The use of this complete, quick-to-use tool with good psychometric properties will allow pregnant women to take stock of their situation, assess whether they have the necessary resources in the psychological and social sphere, and work together with midwives and other health professionals in the areas that are lacking.

Coping strategies during pregnancy and their relationship with anxiety and depression.

How individuals perceive and cope with stressful situations may determine their level of anxiety or depression. The identification of coping strategies (CS) in pregnancy could help prevent depression and anxiety (D&A), and their consequent effects on the health of the mother and the baby. A cross-sectional descriptive correlational study was conducted to identify the CS most commonly used by pregnant women in a Spanish population and to evaluate the association of these CS with D&A. A consecutive sample of 282 pregnant women over 18 years of age were recruited when attended midwife consultations and through snowball sampling between December 2019 and January 2021 in the Basque public health system. CS were measured using the RevisedPrenatal Coping Inventory (NuPCI) questionnaire, assigning the score to an avoidant, preparatory or spiritual scale. Cutoff points were established to categorize anxiety and depressive symptomatology, using the STAI-S and EPDS scales. Multivariate logistic regression models were constructed to analyze the association between CS and D&A. The results show that the higher the score on the avoidance subscale, the higher the likelihood of having an anxiety disorder (OR: 8.88  (95 percent Confidence Interval [CI] 4.26-20.1), and depressive symptoms (OR: 8.29 (95 percent CI 4.24-17.4). Multiparous women are more likely to have anxiety (OR: 3.41  (95 percent CI 1.58-7.5) or depressive symptomatology (OR: 4.1 (95 percent CI 2.04-8.53) during pregnancy. These results highlight the need to consider the evaluation of CS used during pregnancy to tailor the care provided, but further studies on the implementation and effectiveness of interventions are needed.

Non-invasive preimplantation genetic testing for aneuploidies: an update.

Aneuploidy is common among preimplantation human embryos used in assisted reproductive technology. Because abnormal chromosome number can negatively affect reproductive outcome, in-vitro-fertilized embryos routinely undergo aneuploidy testing before transfer into the uterus. This testing typically involves an invasive trophectoderm biopsy of a blastocyst-stage embryo. However, emerging evidence indicates that, during in-vitro development, embryos secrete cell-free DNA into their culture medium; this phenomenon suggests the potential for an alternative, non-invasive assay for aneuploidy. Embryonic cell-free DNA-based assays exhibit high concordance with trophectoderm biopsies, inner cell mass and the whole blastocyst. Yet informativity and concordance rates may be influenced by several factors: the culture day when the medium is collected, contamination with external and/or cumulus cell DNA, and previous manipulation of the embryos. This review discusses non-invasive embryonic cell-free DNA analysis as a biomarker to prioritize blastocysts for transfer to help increase implantation rates and reduce miscarriage rates and time to achieve pregnancy. Ongoing research on the mechanisms underlying embryonic cell-free DNA secretion and how this impacts its role as a biomarker of aneuploidy are also discussed.

Attitudes toward medicalization in childbirth and their relationship with locus of control and coping in a Spanish population.

The dominant model of childbirth in most Western countries is medicalized childbirth. Women's beliefs about whether childbirth should be a medicalized process to a greater or lesser degree may be related, in addition to contextual factors, to internal factors. The objective of the study is to find out if women's locus of control (LC) and stress coping strategies (CS) are related to having a more favourable or less favourable attitude towards medicalization (ATMC). A cross-sectional study was carried out with the participation of 248 women recruited in primary care centres by their midwives. All the women filled in answers on a mobile phone app with various different measurement instruments: the questionnaire created by Benyamini to evaluate their ATMC; the Spanish version of the Wallston MLC to evaluate their LC; and the Spanish adaptation of the "Revised Prenatal Coping Inventory (NuPCI)" scale for the assessment of their CS. The women presented a favourable attitude towards medicalization, with a mean ATMC score of 3.42. Both the LC and the CS of women during pregnancy are related to this attitude. Specifically, having an internal LC and using preparative CS both lower the probability of presenting a favourable attitude towards medicalization, while the lack of a paid job raises the probability. For each point in internal locus and preparatory coping, the ATMC score decreased by 0.02 and 0.23 points, respectively, while it increased by 0.18 for not having a paid job. The influence of these psychological factors must be taken into account in the development of content and interventions that promote a more natural birth.

[Effectiveness of the vagino-rectal exudate self-sampling for prenatal screening of Streptococcus agalactiae infection. GALL study]. / Efectividad de la autotoma del exudado vaginorrectal para el cribado prenatal de la infección por Streptococcus agalactiae. Estudio GALL.

OBJETIVE: To evaluate the effectiveness of vagino-rectal swab autotomy for prenatal screening for GBS infection and to identify the barriers and facilitators encountered by the pregnant woman for this intervention. DESIGN: Cross-sectional study of diagnostic tests. PARTICIPANTS AND SITE: A total of 213 pregnant women who attended the primary care midwife's office in 6 health centers of the Basque Health Service/Osakidetza in Bizkaia, who met the inclusion criteria and agreed to participate in the study, participated in the study. MAIN MEASUREMENTS: The result of the vagino-rectal culture obtained by the pregnant woman was compared with the result of the vagino-rectal culture taken by the midwife in consultation on the same day, and the barriers and facilitators encountered by the women in the self-test were collected. RESULTS: Self-testing as a test for GBS was found to have a sensitivity of 93.3% (95% CI 78.7-98.2), a specificity of 99.4% (95% CI 96.5-99.9), a positive predictive value of 96% (95% CI 82.8-99.4) and a negative predictive value of 98.8% (95% CI 95.6-99.7). 27.3% of respondents encountered some difficulty in the collection, only 4.8% did not feel qualified, 84.2% felt comfortable, 99.5% considered the information provided to be adequate and complete, 94.7% did not find the steps to follow complicated, and 96% were satisfied with the study. CONCLUSIONS: Self-collection of vagino-rectal exudate for GBS detection has proved to be valid and reliable, which would make it possible to offer this option to pregnant women in the systematic screening for GBS infection.

Multicenter prospective study of concordance between embryonic cell-free DNA and trophectoderm biopsies from 1301 human blastocysts.

BACKGROUND: The recent identification of embryonic cell-free DNA in spent blastocyst media has opened a new era of possibilities for noninvasive embryo aneuploidy testing in assisted reproductive technologies. Yet, previous studies assessing a limited number of embryos reported variable concordance between embryonic cell-free DNA and trophectoderm biopsies, thus questioning the validity of this approach. OBJECTIVE: This study aimed to evaluate the concordance and reproducibility of testing embryonic cell-free DNA vs trophectoderm DNA obtained from the same embryo in a large sample of human blastocysts and to assess the contribution of the inner cell mass and trophectoderm to embryonic cell-free DNA released to the culture media. STUDY DESIGN: This is an interim analysis of a prospective, observational study among 8 in vitro fertilization centers in 4 continents to assess consistency between noninvasive embryo aneuploidy testing of embryonic cell-free DNA and conventional trophectoderm biopsy. The analysis included 1301 day-6/7 blastocysts obtained in 406 in vitro fertilization cycles from 371 patients aged 20-44 years undergoing preimplantation genetic testing for aneuploidy. Fresh oocytes underwent intracytoplasmic sperm injection or in vitro fertilization. No previous assisted hatching or vitrification was allowed before media collection. Individual spent blastocyst medium was collected from embryos cultured at least 40 hours from day 4. After media collection, conventional preimplantation genetic testing for aneuploidy, comprising trophectoderm biopsy and blastocyst vitrification, was performed. Embryonic cell-free DNA was analyzed blindly after embryo transfer. Inner cell mass and trophectoderm biopsies were also performed in a subset of 81 aneuploid blastocysts donated for research. RESULTS: Embryonic cell-free DNA analyses were 78.2% (866/1108) concordant with the corresponding trophectoderm biopsies. No significant differences were detected among centers ranging from 72.5% to 86.3%. Concordance rates exceeded 86% when all defined steps in the culture laboratory were controlled to minimize the impact of maternal and operator contamination. Sensitivity per center ranged from 76.5% to 91.3% and specificity from 64.7% to 93.3%. The false-negative rate was 8.3% (92/1108), and false-positive rate was 12.4% (137/1108). The 2 fertilization techniques provided similar sensitivity (80.9% vs 87.9%) and specificity (78.6% vs 69.9%). Multivariate analysis did not reveal any bias from patient clinical background, ovarian stimulation protocols, culture conditions, or embryo quality on testing accuracy of concordance. Moreover, concordances of embryonic cell-free DNA with trophectoderm and inner cell mass suggest that the embryonic cell-free DNA originates from both compartments of the human embryo. CONCLUSION: Noninvasive analysis of embryonic cell-free DNA in spent blastocyst culture media demonstrates high concordance with trophectoderm biopsy results in this large multicenter series. A noninvasive approach for prioritizing embryo euploidy offers important advantages such as avoiding invasive embryo biopsy and decreased cost, potentially increasing accessibility for a wider patient population.

Development of two instruments for assessing maternity health needs: protocol of a clinimetric study.

BACKGROUND: There is an unquestionable need to adapt health care to the needs of each woman, to foster her self-confidence and provide her with the autonomy to manage her own maternity. This involves empowering her to choose and face her model of childbirth and childcare responsibly. The range of self-management health needs tests offered by the scientific community at this stage of life is practically non-existent. In this project, we intend to develop and evaluate the validity, reliability and ease of use of two self-administered analysis instruments for: 1.- Needs of women preparing for childbirth and 2.- Identification of alarm symptoms in the puerperium. METHODS: This is a descriptive study of the clinimetric characteristics and usability of two developed self-applied digital instruments for measuring needs in childbirth and postpartum based on the recommendations made in the consensus-based standards for the selection of health measurement instruments (COSMIN) and by the International Test Commission (ITC). The study consists of two phases: 1 - Evaluation of the clinimetric properties of the two instruments, which were developed and then altered, based on their comprehensibility and global usability estimated from a pilot study and 2 - Pre-implementation study. DISCUSSION: The final product will be two valid, reliable, usable instruments for self-assessment of health needs that are highly acceptable to young couples and the professionals who serve them. They will be a valuable resource for meeting the needs of the population more efficiently and guiding decision-making, and they will contribute to the greater sustainability of the health system.

Clinical application of embryo aneuploidy testing by next-generation sequencing.

We review here the evolution in the field of embryo aneuploidy testing over the last 20 years, from the analysis of a subset of chromosomes by fluorescence in situ hybridisation to the transition toward a more comprehensive analysis of all 24 chromosomes. This current comprehensive aneuploidy testing most commonly employs next-generation sequencing (NGS). We present our experience in over 130 000 embryo biopsies using this technology. The incidence of aneuploidy was lower in trophectoderm biopsies compared to cleavage-stage biopsies. We also confirmed by NGS that embryo aneuploidy rates increased with increasing maternal age, mostly attributable to an increase in complex aneuploid embryos. In contrast, the number of MII oocytes retrieved or the use of oocyte vitrification did not affect aneuploidy rates. Similarly, neither maternal age, oocyte number, nor oocyte vitrification affected the incidence of mosaicism. Analysis of clinical outcomes, indications, and potential benefits of embryo aneuploidy testing revealed advanced maternal age as the most favored group, with some evidence of improved delivery rate per transfer as well as decreased miscarriage rates and time to pregnancy. Other indications are: recurrent miscarriage, repetitive implantation failure, severe male factor, previous trisomic pregnancy, and good prognosis patients mainly undergoing single embryo transfer, with the latter indication used to reduce the occurrence of multiple pregnancies without compromising cycle outcome. In conclusion, NGS has become the most appropriate technology for aneuploidy testing in trophectoderm biopsies, with accurate results, high throughput, and cost efficiency. This technology can be also applied to the analysis of the embryonic cell free DNA released to the culture media at blastocyst stage. This is a promising approach towards a non-invasive preimplantation genetic testing of aneuploidy.

Consensus on priorities in maternal education: results of Delphi and nominal group technique approaches.

BACKGROUND: Maternal education is wide-ranging and covers many areas from pregnancy to the immediate postpartum period and childrearing. However, for it to be effective, more resources need to be assigned to key topics. The goal of this study was to identify and prioritize the most important issues in maternal education, so that specific objectives could subsequently be set and learning outcomes evaluated. METHODS: We drew up a comprehensive list of topics addressed in existing maternal education programs, based on a systematic review of information obtained from the Internet and the experience of the research team. The topics were presented to a multidisciplinary panel whose members were asked to rate them from 1 to 9, and consensus of opinion was reached using a two-round Delphi survey, with consensus defined beforehand as 80% agreement among panelists in awarding a score of 7, 8 or 9. The most highly-rated topics were then discussed and again prioritized by a multidisciplinary team of healthcare and non-healthcare experts, using a nominal group technique. RESULTS: Initially, 650 topics were identified and grouped into 80 categories which were then prioritized by 54 healthcare and non-healthcare experts using a Delphi survey with a study participation rate of around 20%. 63 topics were considered very important, so criteria were restricted and only the 24 highest-scoring selected (95% of agreement on scores ≥7 or 80% of agreement on scores ≥8). Using the nominal group technique, a group of 12 experts identified the following priorities: initiation and establishment of breastfeeding, development of a birth plan, identification of problems and self-care postpartum, nutrition and a healthy lifestyle, options for pain management in labor and birth and characteristics of a normal newborn/looking after a newborn baby. CONCLUSION: This study, with a Delphi study and the Consensus among Experts: the nominal group technique, has succeeded in identifying priority topics in maternal education. We need to assess women's needs in relation to these topics, design an intervention to respond to these needs and evaluate its effectiveness.

Framework for the establishment of a feasible, tailored and effective perinatal education programme.

BACKGROUND: Antenatal education needs to be renewed and adapted to the needs of women. OBJECTIVES: to assess women needs, identify factors that influence the desired outcomes, and propose a framework for developing new perinatal education based on the guidance published by the UK Medical Research Council for the development and evaluation of complex interventions in primary care. METHODS: For this study: (a) four focus group sessions were held from October to November 2010 in Bizkaia (Spain) with 30 women exploring their needs during pregnancy and postpartum; (b) two literature reviews were conducted on women's needs at these times and theoretical models of healthcare education; and (c) seven discussion and consensus sessions were run with a group of experts composed of midwifes, gynaecologists, paediatricians, and paediatric and postpartum nurses. RESULTS: Various areas for improvement were identified: needs assessment of each woman/family, consideration of pregnancy and childbirth as normal physiological processes, participation of fathers, establishment of social networks, continuity of postpartum care, better access to and training for midwives, and more flexible format and contents for the programme. We propose a woman-focused framework that includes three exploratory interviews during pregnancy, personalized interventions coordinated between professionals, empowerment to choose the type of birth, and postpartum activities. CONCLUSION: New perinatal education should be on-going and focused on each woman. It is necessary to assess the feasibility of implementing this type of programme, depending on the context, professionals' readiness for change and characteristics of the proposed interventions. Then, its effectiveness and sustainability must be assessed.

[Perceived needs of women regarding maternity. Qualitative study to redesign maternal education]. / Necesidades percibidas por las mujeres respecto a su maternidad. Estudio cualitativo para el rediseño de la educación maternal.

OBJECTIVES: To assess women's perceptions of their needs during the process of becoming a mother and identify what they want from maternal education. DESIGN: Qualitative study with focus groups. SETTING: Bizkaia health region, Basque Health Service (Osakidetza), Spain. PARTICIPANTS: Thirty one women were recruited consecutively by midwives at six Osakidetza health centres. METHODS: Four sessions were held from September to November 2010 in Bizkaia (Spain), the four groups being stratified by socioeconomic status and stage of the process (pregnancy vs. postnatal period). To collate the information related to the various topics discussed, we used manifest content analysis that was facilitated by use of ATLAS.ti software. RESULTS: The focus of the women worries changes over time. In early pregnancy, women's main concern was for "everything to go well". As the pregnancy progressed, they needed more emotional support and wanted to feel confident and be self-reliant to face their fears of the birth and care for their child. They needed greater accompaniment in the puerperium and less pressure concerning breastfeeding. They also wanted an extended programme of perinatal rather than just antenatal education, which was more participatory and flexible and greater participation of their partner. CONCLUSION: Women have the same social and family networks needs, regardless of cultural differences between Anglo-Saxon and Southern European countries. We recommend an perinatal education to empower women to manage their own health and that of their family and link the health system with other networks of personal and social support for women.

Dopamine agonist inhibits vascular endothelial growth factor protein production and secretion in granulosa cells.

BACKGROUND: Dopamine receptor 2 agonists (D2-ags) inhibit vascular endothelial growth factor (VEGF) secretion in luteinized granulosa cells (LGCs) both in vitro and in vivo. However, the mechanism of D2 regulation of the VEGF/VEGF Receptor 2 (VEGFR-2) pathway remains to be elucidated. We sought to determine the effects of D2 signaling on VEGF transcription and translation in LGCs, with the expectation of identifying potential D2-ag-based therapies for ovarian hyperstimulation syndrome (OHSS). FINDINGS: LGCs from egg donors were cultured with chorionic gonadotropin (hCG) in the presence of Actinomycin-D (ActD) or Brefeldin-A (BFA) to evaluate the effects of a D2-ag, cabergoline (Cb2), on VEGF secretion. The contribution of the conventional Gi/Go, Gz and AKT/ß-Arrestin pathways in the VEGF regulation was assessed by adding pertussis toxin (PTX), phorbol 12-myristate 13-acetate (PMA), or wortmannin (WT). While Cb2 inhibited VEGF secretion by interfering with VEGF peptide translation and secretion, inhibition of conventional D2 transduction pathways did not reverse Cb2-mediated inhibition of VEGF secretion. CONCLUSIONS: The effects of D2-ag on VEGF translation and secretion are mediated by D2 signaling pathways that have yet to be described. We found that D2-ag inhibits VEGF secretion at the post-transcriptional level, suggesting that D2-ag treatment should be combined with therapies that inhibit VEGF transcription, such as the employment of LH or GnRH for triggering ovulation, to improve the efficacy of OHSS prevention.

Anxiety and depression in pregnancy: associated variables during the COVID-19 pandemic period.

OBJECTIVE: To assess the association between age, parity, nationality, educational level, employment and partner support with depression and anxiety during pregnancy. To determine the impact of the evolution of the COVID-19 pandemic on the anxiety and depression levels of the participants. METHOD: Cross-sectional descriptive correlational study carried out in the Osakidetza/Basque Health Service, Primary Care, Bizkaia (Spain). A total of 295 pregnant women between 8 and 41 weeks participated. The association between age, parity, nationality, educational level, employment, partner support and COVID-19 incidence rates during the study period and depression (measured with the Edinburgh Postnatal Depression Scale) and anxiety (measured with the State-Anxiety Inventory/STAI-S subscale) scores during pregnancy was measured. A logistic regression model was constructed and odds ratios and their 95% confidence intervals were estimated. RESULTS: The mean score was 18.75 points for anxiety (SD=8.69) and 6.45 points for depression (SD=4.32). Women expecting their second or later child had higher scores for depression (OR 2.51[95%IC: 1.26-5.01)) and anxiety (OR 1.98 [95%IC: 1.01-3.89]). Having completed university studies was associated with lower scores in depression (OR 0.28 [95%IC: 0.11-0.77]) and in anxiety (OR 0,2 [95%IC: 0.08-0.54]). A good relationship with the partner was associated with lower levels of anxiety (OR 0.45 [95%IC: 0.24-0.81]). The mean anxiety and depression scores are related to the incidence of COVID-19 cases; the anxiety score was significantly higher in periods of higher incidence. CONCLUSIONS: Greater emotional care for pregnant women with low educational level, multiparous and with weak support from their partner, would reduce anxiety and depression in pregnancy. Health emergency situations affect mental health during gestation.

Culture time to optimize embryo cell-free DNA analysis for frozen-thawed blastocysts undergoing noninvasive preimplantation genetic testing for aneuploidy.

OBJECTIVE: To investigate the ideal time in culture to optimize embryo cell-free deoxyribonucleic acid (cfDNA) analysis in frozen-thawed blastocysts undergoing noninvasive preimplantation genetic testing for aneuploidy (PGT-A). Cell-free DNA is released into the spent blastocyst media (spent media) by the embryo. However, the optimal timing to determine maximal cfDNA in the case of frozen-thawed blastocysts undergoing noninvasive PGT-A remains to be elucidated. DESIGN: In this prospective observational study, 135 spent media and corresponding whole blastocysts were collected from January 2021 through March 2022. SETTING: Private fertility clinics. PATIENTS: Day-5 frozen-thawed blastocysts were cultured for 8 hours (Day-5 Short) or 24 hours (Day-5 Long), whereas day-6 frozen-thawed blastocysts were cultured for 8 hours (Day-6 Short). The spent media and whole blastocysts were then collected for further analysis. Spent media and whole blastocysts were amplified using whole genome amplification and sequenced using next-generation sequencing. MAIN OUTCOME MEASURES: Informativity and concordance rates between cfDNA in spent media and whole blastocyst DNA were compared according to the different times in culture. RESULTS: When comparing time in culture, informativity rates for spent media were significantly higher for Day-5 Long and Day-6 Short (>91%) compared with the Day-5 Short group (<60%). A similar trend was observed for cases with and without a previous PGT-A. Regarding blastocyst expansion grade, informativity rates were lower on Day-5 Short compared with Day-5 Long and Day-6 Short, regardless of expansion degree. This decrease was significant for Gardner-grade expansion grades 3, 4, and 5-6. In addition, for a similar time in culture, the grade of expansion did not have an impact on the informativity rates. For concordance rates, no significant differences were observed among the 3 groups. In all cases, concordance rates were 90.5% for Day-5 Short, 93.6% for Day-5 Long, and 92.3% for Day-6 Short. No impact of the expansion grade was observed on concordance rates. CONCLUSION: Noninvasive PGT-A in frozen-thawed blastocysts yields very high concordance rates with whole blastocysts, possibly limiting the need for invasive PGT-A and making it available for a wider range of patients.

Antenatal education and breastfeeding in a cohort of primiparas.

AIMS: To assess the association between attendance at antenatal education sessions and breastfeeding during the first year of life. BACKGROUND: Although there is evidence that antenatal education encourages breastfeeding, the size and duration of its effect remain unclear. DESIGN: A prospective cohort study. METHODS: The study was conducted in Bizkaia (North of Spain) between May 2005-June 2007 with a consecutive sample of 614 primiparas. Women were classified into three groups according to whether they had received antenatal education and, if so, how many classes (0, 1-4, or 5 or more). Telephone interviews at 1·5, 3, 6 and 12 months were used to estimate the risk of cessation of any breastfeeding and to compare the groups with Cox proportional hazards regression models adjusted for potential confounders. FINDINGS: Initially, 90% of women breastfed their infants, with no differences between the groups. During the first month, the risk of cessation of any breastfeeding was three times as high among non-attendees and twice as high among women who attended 1-4 classes compared with those who attended 5 or more classes. The risk was, however, similar in the three groups from the end of first month onwards. CONCLUSION: The results suggest that though antenatal education may be associated with higher rates of breastfeeding in our setting this is only the case for the first month after the birth. Further research is necessary to optimize this beneficial effect to achieve long-term continuation of breastfeeding.

Cell-free deoxyribonucleic acid analysis in preimplantation genetic testing.

Detection of chromosomal aneuploidies and monogenic disorders in preimplantation embryos is essential for selecting the best embryo for transfer during in vitro fertilization to achieve a healthy pregnancy. Preimplantation genetic testing (PGT) is typically performed on preimplantation embryos to select a genetically normal embryo for transfer. A trophectoderm biopsy is necessary for PGT; this is an invasive procedure to the embryo that requires specialized equipment and highly trained embryologists, resulting in high costs associated with in vitro fertilization treatment. Moreover, the biopsy procedure may increase the likelihood of developing pregnancy complications, such as preeclampsia and hypertensive disorders. Therefore, there is a need for noninvasive embryo screening strategies. The presence of cell-free deoxyribonucleic acid in the embryo culture medium presents an opportunity to screen for genetic abnormalities. Cell-free deoxyribonucleic acid is released by embryos in the latter stages of preimplantation development, and its analysis has been proposed as a noninvasive approach for PGT. Here, we review studies reporting the concordance rates between cell-free deoxyribonucleic acid and trophectoderm biopsies, or whole blastocysts, in couples undergoing PGT. Noninvasive PGT results are promising for aneuploidy detection, with some early evidence of successful clinical application. Further research is required to explore its application for the detection of structural rearrangements and monogenic disorders.

Efficiency and purity provided by the existing methods for the isolation of luteinized granulosa cells: a comparative study.

BACKGROUND: Several protocols for the isolation of luteinized granulosa cells (LGCs) contained in follicular fluid have been described but no previously published study has compared the relative efficiency of these protocols. Our objective is to obtain conclusive scientific evidence for the superiority of one method over another. METHODS: Different purification methods for LGCs based on the recognition of specific cell markers, aggregates, differential adhesion and LGC size were evaluated. We compared the levels of CD45 cell contamination and the percentage of total cell viability in paired aliquots of cells (before and after purification) derived from the follicular fluid obtained from women who were donating oocytes (n = 72). Each of the six purification methods was performed six times using pooled follicular fluids from two women. RESULTS: Samples processed by means of recognition of specific cell markers were characterized by their greater purity (0.1-1.33% CD45+) but low rate of LGC recovery (17.13-25.4%) when compared with the other methods (3.29-12% CD45+, P < 0.05 and 51.67-73.20% LGC, P < 0.05). It is noteworthy that the filter method, which is based on the LGC size, combined one of the highest rates of LGC recovery (∼70%) with acceptable low levels of contamination (<5%). CONCLUSIONS: There is currently no gold standard method for the isolation of LGCs, and protocols should be chosen depending on the purpose in question. We conclude that fluorescence-activated cell sorting is the best protocol for isolating LGCs when purity is the principal criterion, and magnetic separation when both purity and viability are essential. However, cell straining (filter) is probably the least laborious and, overall, the most efficient method to isolate LGCs.