Preimplantation genetic diagnosis (PGD) for monogenic disorders currently involves polymerase chain reaction (PCR)-based methods, which must be robust, sensitive and highly accurate, precluding misdiagnosis. Twelve adverse misdiagnoses reported to the ESHRE PGD-Consortium are likely an underestimate. This retrospective study, involving six PGD centres, assessed the validity of PCR-based PGD through reanalysis of untransferred embryos from monogenic-PGD cycles. Data were collected on the genotype concordance at PGD and follow-up from 940 untransferred embryos, including details on the parameters of PGD cycles: category of monogenic disease, embryo morphology, embryo biopsy and genotype assay strategy. To determine the validity of PCR-based PGD, the sensitivity (Se), specificity (Sp) and diagnostic accuracy were calculated. Stratified analyses were also conducted to assess the influence of the parameters above on the validity of PCR-based PGD. The analysis of overall data showed that 93.7% of embryos had been correctly classified at the time of PGD, with Se of 99.2% and Sp of 80.9%. The stratified analyses found that diagnostic accuracy is statistically significantly higher when PGD is performed on two cells versus one cell (P ¼ 0.001). Se was significantly higher when multiplex protocols versus singleplex protocols were applied (P ¼ 0.005), as well as for PGD applied on cells from good compared with poor morphology embryos (P ¼ 0.032). Morphology, however, did not affect diagnostic accuracy. Multiplex PCR-based methods on one cell, are as robust as those on two cells regarding false negative rate, which is the most important criteria for clinical PGD applications. Overall, this study demonstrates the validity, robustness and high diagnostic value of PCR-based PGD.

IntroductionOverweight and obese women may require higher doses of gonadotrophin when undergoing In Vitro Fertilization Treatment (IVF). Consequently, one may expect a sub-optimal oocyte retrieval in the first treatment cycle and thus a larger compensation in gonadotrophin-dose in the following treatment-cycles and a more favorable outcome. The main objective was to explore if treatment cycle number modifies the outcome when investigating the effect of female Body Mass Index (BMI) on oocyte quantity in IVF.Material and MethodsA historical cohort study was conducted on 5,342 treatment-cycles during the period 1999–2009. Exclusion criteria were missing information on BMI or treatment type. Further, women were excluded if they had ovulated before oocyte retrieval. According to baseline BMI, women were divided into four categories following the World Health Organization standards. Multiple linear regressions analyses were performed accounting for the non-independence of ≥2 cycles in a woman.ResultsStratification according to cycle number revealed a more suboptimal outcome in the first treatment- cycles than in the following cycles, suggesting a possible interaction or effect modification from cycle number or a factor related to cycle number. The median dose of total follicular stimulating hormone given to the four BMI groups could not straight forwardly explain the less optimal oocyte outcome observed in first treatment cycles. No statistically significant differences were observed in oocyte yield for underweight, overweight and obesity compared to normal weight women when analyzing all treatment-cycles. Overweight women had significantly fewer mature (MII) oocytes (p = 0.009) than normal weight women, whereas no differences was observed for underweight and obese women.ConclusionOur study suggests a possible interaction or effect modification related to treatment cycle number. Investigating the effects of BMI on IVF-results in first treatment-cycles alone should be carried out cautiously.

STUDY QUESTION Do young women with early ovarian ageing (EOA), defined as unexplained, and repeatedly few oocytes harvested in ART have an increased risk of age-related events? SUMMARY ANSWER At follow-up, women with idiopathic EOA had an increased risk of age-related events compared to women with normal ovarian ageing (NOA). WHAT IS KNOWN ALREADY Early and premature menopause is associated with an increased risk of cardiovascular diseases (CVDs), osteoporosis and death. In young women, repeated harvest of few oocytes in well-stimulated ART cycles is a likely predictor of advanced menopausal age and may thus serve as an early marker of accelerated general ageing. STUDY DESIGN, SIZE, DURATION A register-based national, historical cohort study. Young women (≤37 years) having their first ART treatment in a public or private fertility clinic during the period 1995–2014 were divided into two groups depending on ovarian reserve status: EOA (n = 1222) and NOA (n = 16 385). Several national registers were applied to assess morbidity and mortality. PARTICIPANTS/MATERIALS, SETTING, METHODS EOA was defined as ≤5 oocytes harvested in a minimum of two FSH-stimulated cycles and NOA as ≥8 oocytes in at least one cycle. Cases with known causes influencing the ovarian reserve (endometriosis, ovarian surgery, polycystic ovary syndrome, chemotherapy etc.) were excluded. To investigate for early signs of ageing, primary outcome was an overall risk of ageing-related events, defined as a diagnosis of either CVD, osteoporosis, type 2 diabetes, cancer, cataract, Alzheimer’s or Parkinson’s disease, by death of any-cause as well as a Charlson comorbidity index score of ≥1 or by registration of early retirement benefit. Cox regression models were used to assess the risk of these events. Exposure status was defined 1 year after the first ART cycle to assure reliable classification, and time-to-event was measured from that time point. MAIN RESULTS AND THE ROLE OF CHANCE Median follow-up time from baseline to first event was 4.9 years (10/90 percentile 0.7/11.8) and 6.4 years (1.1/13.3) in the EOA and NOA group, respectively. Women with EOA had an increased risk of ageing-related events when compared to women with a normal oocyte yield (adjusted hazard ratio 1.24, 95% CI 1.08 to 1.43). Stratifying on categories, the EOA group had a significantly increased risk for CVD (1.44, 1.19 to 1.75) and osteoporosis (2.45, 1.59 to 3.90). Charlson comorbidity index (1.15, 0.93 to 1.41) and early retirement benefit (1.21, 0.80 to 1.83) was also increased, although not reaching statistical significance. LIMITATIONS, REASONS FOR CAUTION Cycles never reaching oocyte aspiration were left out of account in the inclusion process and we may therefore have missed women with the most severe forms of EOA. We had no information on the total doses of gonadotrophin administered in each cycle. WIDER IMPLICATIONS OF THE FINDINGS These findings indicate that oocyte yield may serve as marker of later accelerated ageing when, unexpectedly, repeatedly few oocytes are harvested in young women. Counselling on life-style factors as a prophylactic effort against cardiovascular and other age-related diseases may be essential for this group of women. STUDY FUNDING/COMPETING INTEREST(S) No external funding was received for this study. All authors declare no conflict of interest. TRIAL REGISTRATION NUMBER N/A

During the second wave of the COVID-19 pandemic, despite a strong focus on preventive measures, healthcare workers were still at increased risk of SARS-CoV-2 infection following close contact with patients and co-workers as well as persons outside work with COVID-19. Intensified preventive actions to secure healthcare workers' health during ongoing and future waves of SARS-CoV-2 and other infections are thus needed.

Bone marrow transplantation may be life saving in cases of hematopoietic disease, severe congenital immunodeficiency or malignancy. An HLA-matching sibling often gives the best success, but this may not be an option, nor may an HLA-matching unrelated donor be found. Preimplantation genetic diagnosis with HLA-matching embryos may then be a solution. We report the first Danish child born after such diagnosis with identification of healthy embryos and HLA matching for a sibling with chronic granulomatous disease. Our patient had 13 in vitro fertilization (IVF) cycles; 286 oocytes were collected and 74 embryos analysed. Sixteen of these (22%) were either healthy or carriers. Five embryos were transferred in four stimulated fresh IVF cycles. Four embryos were frozen, and two were later transferred. Two successive clinical pregnancies ensued. In the first, prenatal diagnosis revealed trisomy 21, and the fetus was aborted. In the second pregnancy, chorionic villus sampling revealed a normal karyotype, the diagnosis was confirmed, and the pregnancy was normal. At delivery, 82 mL of cord blood was collected for later transplantation to the diseased sibling.

ObjectivesTo assess if healthcare workers during the second wave of the COVID-19 pandemic had increased SARS-CoV-2 infection rates following close contact with patients, co-workers and persons outside work with COVID-19.DesignProspective cohort study.SettingPublic hospital employees in Denmark.Participants5985 healthcare workers (88.6% women) who daily on a smartphone reported COVID-19 contact.Main outcome measuresSARS-CoV-2 infection rates defined by the first positive polymerase chain reaction (PCR) test recorded in a register with complete test coverage.Results159 positive and 35 996 negative PCR tests for SARS-CoV-2 were recorded during 514 165 person-days of follow-up November 25, 2020 - April 30, 2021. The SARS-CoV-2 infection rate for healthcare workers who during the previous 3-7 days had close contact with COVID-19 patients was 153.7 per 100 000 person-days (0.15% per day) corresponding with an incidence rate ratio of 3.17 (40 cases, 95% CI 2.15 to 4.66) when compared with no close contact with COVID-19 patients. SARS-CoV-2 incidence rate ratios following close contact with co-workers and persons outside work with COVID-19 were 2.54 (10 cases, 95% CI 1.30 to 4.96) and 17.79 (35 cases, 95% CI 12.05 to 26.28). These estimates were mutually adjusted and further adjusted for age, sex, month and number of previous PCR tests.ConclusionsDespite strong focus on preventive actions during the second wave of the pandemic, healthcare workers were still at increased risk of SARS-CoV-2 infection when in close contact with patients with COVID-19. The numbers affected were comparable to the numbers affected following COVID-19 contact outside work.Close contact with co-workers was also a risk factor. This stresses the need for increased focus on preventive actions to secure healthcare workers’ health during ongoing and future waves of SARS-CoV-2 and other infections.Summary boxWhat is already known on this topic?During the first wave of the COVID-19 pandemic, preventive measures were inadequate and healthcare workers were at increased risk of SARS-CoV-2 infection.What this study adds?During the second wave of the pandemic, despite a strong focus on preventive actions, healthcare workers were still at increased risk of SARS-CoV-2 infection following close contact with COVID-19 patients.The numbers affected were comparable to the numbers affected following COVID-19 contact outside work.Healthcare works were also at increased risk following close contact with co-workers.

Purpose To evaluate whether young women with idiopathic early ovarian aging, as defined by producing fewer oocytes than expected for a given age over multiple in vitro fertilization (IVF) cycles, have changes in telomere length and epigenetic age indicating accelerated biological aging (i.e., increased risk of morbidity and mortality). Methods A prospective cohort study was conducted at two Danish public fertility clinics. A total of 55 young women (≤ 37 years) with at least two IVF cycles with ≤ 5 harvested oocytes despite sufficient stimulation with follicle-stimulating hormone (FSH) were included in the early ovarian aging group. As controls, 52 young women (≤ 37 years) with normal ovarian function, defined by at least eight harvested oocytes, were included. Relative telomere length (rTL) and epigenetic age acceleration (AgeAccel) were measured in white blood cells as markers of premenopausal accelerated biological aging. Results rTL was comparable with a mean of 0.46 (± SD 0.12) in the early ovarian aging group and 0.47 (0.14) in the normal ovarian aging group. The AgeAccel of the early ovarian aging group was, insignificantly, 0.5 years older, but this difference disappeared when adjusting for chronological age. Sub-analysis using Anti-Müllerian hormone (AMH) as selection criterion for the two groups did not change the results. ConclusionWe did not find any indications of accelerated aging in whole blood from young women with idiopathic early ovarian aging. Further investigations in a similar cohort of premenopausal women or other tissues are needed to fully elucidate the potential relationship between premenopausal accelerated biological aging and early ovarian aging.

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