DNA methylation differences at birth after conception through ART.

STUDY QUESTION: Is there a relation between ART and DNA methylation (DNAm) patterns in cord blood, including any differences between IVF and ICSI? SUMMARY ANSWER: DNAm at 19 CpGs was associated with conception via ART, with no difference found between IVF and ICSI. WHAT IS KNOWN ALREADY: Prior studies on either IVF or ICSI show conflicting outcomes, as both widespread effects on DNAm and highly localized associations have been reported. No study on both IVF and ICSI and genome-wide neonatal DNAm has been performed. STUDY DESIGN, SIZE, DURATION: This was a cross-sectional study comprising 87 infants conceived with IVF or ICSI and 70 conceived following medically unassisted conception. The requirement for inclusion in the study was an understanding of the Swedish language and exclusion was the use of donor gametes. PARTICIPANTS/MATERIALS, SETTING, METHODS: Participants were from the UppstART study, which was recruited from fertility and reproductive health clinics, and the Born into Life cohort, which is recruited from the larger LifeGene study. We measured DNAm from DNA extracted from cord blood collected at birth using a micro-array (450k array). Group differences in DNAm at individual CpG dinucleotides (CpGs) were determined using robust linear models and post-hoc Tukey's tests. MAIN RESULTS AND THE ROLE OF CHANCE: We found no association of ART conception with global methylation levels, imprinted loci and meta-stable epialleles. In contrast, we identify 19 CpGs at which DNAm was associated with being conceived via ART (effect estimates: 0.5-4.9%, PFDR < 0.05), but no difference was found between IVF and ICSI. The associated CpGs map to genes related to brain function/development or genes connected to the plethora of conditions linked to subfertility, but functional annotation did not point to any likely functional consequences. LIMITATIONS, REASONS FOR CAUTION: We measured DNAm in cord blood and not at later ages or in other tissues. Given the number of tests performed, our study power is limited and the findings need to be replicated in an independent study. WIDER IMPLICATIONS OF THE FINDINGS: We find that ART is associated with DNAm differences in cord blood when compared to non-ART samples, but these differences are limited in number and effect size and have unknown functional consequences in adult blood. We did not find indications of differences between IVF and ICSI. STUDY FUNDING/COMPETING INTEREST(S): E.W.T. was supported by a VENI grant from the Netherlands Organization for Scientific Research (91617128) and JPI-H2020 Joint Programming Initiative a Healthy Diet for a Healthy Life (JPI HDHL) under proposal number 655 (PREcisE Project) through ZonMw (529051023). Financial support was provided from the European Union's Seventh Framework Program IDEAL (259679), the Swedish Research Council (K2011-69X-21871-01-6, 2011-3060, 2015-02434 and 2018-02640) and the Strategic Research Program in Epidemiology Young Scholar Awards, Karolinska Institute (to A.N.I.) and through the Swedish Initiative for Research on Microdata in the Social And Medical Sciences (SIMSAM) framework grant no 340-2013-5867, grants provided by the Stockholm County Council (ALF-projects), the Strategic Research Program in Epidemiology at Karolinska Institutet and the Swedish Heart-Lung Foundation and Danderyd University Hospital (Stockholm, Sweden). The funders had no role in study design, data collection, analysis, decision to publish or preparation of the manuscript. The authors declare no competing interests. TRIAL REGISTRATION NUMBER: N/A.

A prospective investigation of perceived stress, infertility-related stress, and cortisol levels in women undergoing in vitro fertilization: influence on embryo quality and clinical pregnancy rate.

INTRODUCTION: Women undergoing fertility treatment experience high levels of stress. However, it remains uncertain if and how stress influences in vitro fertilization (IVF) cycle outcome. This study aimed to investigate whether self-reported perceived and infertility-related stress and cortisol levels were associated with IVF cycle outcomes. MATERIAL AND METHODS: A prospective cohort of 485 women receiving fertility treatment was recruited from September 2011 to December 2013 and followed until December 2014. Data were collected by online questionnaire prior to IVF start and from clinical charts. Salivary cortisol levels were measured. Associations between stress and cycle outcomes (clinical pregnancy and indicators of oocyte and embryo quality) were measured by logistic or linear regression, adjusted for age, body mass index, education, smoking, alcohol and caffeine consumption, shiftwork and night work. RESULTS: Ultrasound verified pregnancy rate was 26.6% overall per cycle started and 32.9% per embryo transfer. Stress measures were not associated with clinical pregnancy: when compared with the lowest categories, the adjusted odds ratio (OR) and 95% confidence interval (CI) for the highest categories of the perceived stress score was 1.04 (95% CI 0.58-1.87), infertility-related stress score was OR = 1.18 (95% CI 0.56-2.47), morning and evening cortisol was OR = 1.18 (95% CI 0.60-2.29) and OR = 0.66 (95% CI 0.34-1.30), respectively. CONCLUSIONS: Perceived stress, infertility-related stress, and cortisol levels were not associated with IVF cycle outcomes. These findings are potentially reassuring to women undergoing fertility treatment with concerns about the influence of stress on their treatment outcome.

Radionuclide hysterosalpingography is not a reliable tool for investigation of fallopian tube transport--a controlled randomized study using particles of two sizes during three different parts of the menstrual cycle.

BACKGROUND/AIMS: Earlier studies with radionuclide hysterosalpingography (RN-HSG) using either small or large particles have shown different results. Some studies have questioned the clinical relevance of RN-HSG. The aim of this study was to evaluate the RN-HSG method using two different sizes of radio-labelled particles during different parts of the menstrual cycle. METHODS: Ten healthy women, 28-40 years of age (mean 34.5), with proven fertility, regular menstrual cycle, not using oral contraceptives and due to undergo laparoscopic sterilization, were randomized to RN-HSG after deposition of a solution with either small Albures or large MAASOL, (99m)Tc-radio-labelled particles into the cervix uteri. Investigations were, for every patient, performed during the follicular, peri-ovulatory and mid-luteal parts of the menstrual cycle. Transport of radioactivity from cervix to uterus and Fallopian tubes was evaluated. RESULTS: The RN-HSG investigations could not show any difference between the two different sizes of radio-labelled particles. Transport of radioactivity to the Fallopian tube was seen only in some cases and most frequently during the peri-ovulatory period. CONCLUSION: We found the RN-HSG investigations inconclusive. The RN-HSG method does not seem to be a reliable tool for the investigation of Fallopian tube transport.

The Uppsala-Stockholm Assisted Reproductive Techniques (UppStART) study.

PURPOSE: The Uppsala-Stockholm Assisted Reproductive Techniques (UppStART) study is a prospectively recruited sample of couples undergoing assisted reproduction in Stockholm and Uppsala county in Sweden. The study was initiated to (1) investigate possible changes in the epigenetic profile of infants inferred through the ART procedures and their consequence and (2) to assess the impact of lifestyle and health exposures on treatment outcome. PARTICIPANTS: Recruitment took place between September 2011 and December 2013, and in vitro fertilisation (IVF) cycles initiated and pregnancies conceived during this time were followed until December 2014. The cohort includes 971 participants (n= 514 women; n= 457 men), and 129 pregnancies were achieved from the first IVF cycle included in the study. FINDINGS TO DATE: Self-reported demographic, health and lifestyle data were collected from a baseline questionnaire, and to assess changes to lifestyle, a follow-up questionnaire was issued at the time of oocyte retrieval, and at subsequent IVF cycles. Questionnaire data were linked to data extracted from medical records. Biological samples were collected at baseline: blood for extraction of serum, plasma and DNA, morning and evening saliva samples for cortisol measurement and at delivery including samples of maternal blood, placenta and amniotic fluid, and cord blood for epigenetic analysis. FUTURE PLANS: Through the unique identification number assigned to each Swedish citizen at birth or immigration, UppStART study participants will be linked to the Swedish population-based national and quality registers to provide data from prenatal, obstetrical, neonatal and infant care, and subsequent updates will provide data on childhood health and educational outcomes. Collaboration and use of UppStART data is encouraged, and more information about access can be found at www.ki.se/meb/uppstart.

Detection of Tc-99m labelled albumin particles in the Fallopian tubes after intraperitoneal deposition.

OBJECTIVE: To study the Fallopian tube uptake of intra-abdominal albumin particles. STUDY DESIGN: Six healthy women volunteered for ultrasound guided deposition of Tc-99m labelled albumin particles into the pouch of Douglas, before undergoing sterilisation by salpingectomy. RESULTS: On gamma camera images radioactivity spread evenly over the parietal peritoneum. Autoradiography of the Fallopian tubes showed radioactive "hot spots" on the inner tubal surface. A rapid increase in blood radioactivity stabilised after 2-3 h while radioactivity remained high in the peritoneal cavity. Cervical mucus showed a significant radioactivity. CONCLUSION: Autoradiography strongly indicated that Tc-99m labelled particles deposited into the pouch of Douglas were transported to the Fallopian tubes. The small uptake of radioactivity identified in blood was most likely due to a rapid diffusion of free Tc-99m pertechnetate from the peritoneal cavity. The radioactivity identified in cervical mucus could be pertechnetate or labelled albumin particles or a combination of both. Future development of the present technique may be used for fertility investigation.

Emotional adaptation following successful in vitro fertilization.

OBJECTIVE: To assess the emotional impact of infertility after successful IVF and to compare parents who have undergone IVF (IVF parents) and parents who have not undergone IVF (non-IVF parents) regarding parental stress and the marital relationship during the transition to parenthood. DESIGN: A study with qualitative and longitudinal quantitative assessments. SETTING: University IVF clinics and antenatal clinics in Stockholm. PATIENT(S): Fifty-five IVF mothers, 53 IVF fathers, 40 non-IVF mothers, and 36 non-IVF fathers. INTERVENTION(S): IVF parents were interviewed. All subjects completed self-rating scales in early pregnancy and at 2 and 6 months postpartum. MAIN OUTCOME MEASURE(S): Interviews about perception of infertility and scalar measurement of parental stress and the marital relationship. RESULT(S): Negative feelings related to infertility were not easily overcome among the IVF parents. Their levels of stress related to parenthood were similar to those of non-IVF parents, and both groups reported decreased satisfaction with the marital relationship during the transition to parenthood. CONCLUSION(S): The inability to conceive naturally continues to affect the current lives of a proportion of IVF parents. The results suggest that IVF parents may benefit from counseling with regard to the potential long-term impacts of infertility, disclosure issues, and decisions regarding future children. However, levels of parental stress and patterns of partner satisfaction are similar to those of parents with children conceived "naturally."

Severe jaundice in early IVF pregnancy.

Jaundice in early pregnancy after in vitro fertilization (IVF) is extremely rare. We report a case of severe jaundice in an IVF treated patient, with a clinical picture similar to intrahepatic cholestasis of pregnancy (ICP). We suggest strategies to prevent similar cases in the future.

Radioactivity in cervical mucus after intraperitoneal deposition of Tc-99m labelled albumin particles before and after sterilisation.

OBJECTIVE: To study the transport of radiolabelled albumin particles from the pouch of Douglas to cervical mucus. STUDY DESIGN: A prospective randomised study, including 10 healthy women, undergoing laparoscopic sterilization. A solution of human serum albumin particles, labelled with Technetium 99(m) (Tc-99(m)) was injected into the pouch of Douglas, either before or after sterilisation. RESULTS: Gamma camera images showed gradual spread to the entire peritoneal cavity. Blood samples, showed increasing levels of radioactivity. Measurement of radioactivity in cervical mucus showed significant activity both in women sterilized before and after the deposition. CONCLUSION: The study showed evidence of transport of radioactivity from the pouch of Douglas to cervical mucus. The transport seemed to consist of free activity since no radionuclide labelled particles could be detected in cervical mucus. The transport was probably haematogenous or lymphatic and not intraluminal through the fallopian tube. Further studies utilizing particles comparable in size to fertilized ova are required to design methods for evaluating the transport from the fallopian tube to the uterine cavity.

Personality factors and emotional responses to pregnancy among IVF couples in early pregnancy: a comparative study.

BACKGROUND: Little is known as to whether negative experiences associated with infertility and IVF treatment persist after successful treatment. The aim of the study was to compare couples who have conceived after IVF and couples who have conceived naturally regarding personality factors and emotional responses to pregnancy. METHODS: Fifty-seven women pregnant after IVF and 55 male partners and 43 women who had conceived naturally and 39 male partners were recruited from university IVF clinics and antenatal clinics in Stockholm. The subjects were interviewed about their socio-demographic background. They completed scales of personality traits, anxiety, emotional responses to pregnancy, marital adjustment and reactions to recalled infertility while in pregnancy week 13 (range 11-17). RESULTS: The results showed that the IVF women had more muscular tension and were more anxious about loosing the pregnancy than the control women. The IVF women with high infertility distress were more anxious about loosing the pregnancy and less ambivalent than the women with lower distress. The IVF men had more somatic anxiety, indirect aggression, guilt, and were more detached and more anxious about loosing the pregnancy and less ambivalent than the control men. The IVF men with high infertility distress were more anxious about the baby not being normal than the men with lower infertility distress. CONCLUSIONS: The women and men who had conceived after IVF differed on a number of personality dimensions and emotional responses to the pregnancy from that of the women and the men who had conceived naturally. The results suggest that IVF couples may need additional emotional support in early pregnancy.