Aneuploidy is frequent in heterozygous diploid and triploid hydatidiform moles.

Hydatidiform moles are abnormal conceptuses. Many hydatidiform moles are diploid androgenetic, and of these, most are homozygous in all loci. Additionally, most hydatidiform moles are euploid. Using Single Nucleotide Polymorphism (SNP) array analysis, in two studies a higher frequency of aneuploidy was observed in diploid androgenetic heterozygous conceptuses, than in their homozygous counterparts. In the Danish Mole Project, we analyze conceptuses suspected to be hydatidiform moles due to the clinical presentation, using karyotyping and Short Tandem Repeat (STR) analysis. Among 278 diploid androgenetic conceptuses, 226 were homozygous in all loci and 52 (18.7%) were heterozygous in several loci. Among 142 triploid diandric conceptuses, 141 were heterozygous for paternally inherited alleles in several loci. Here we show that the frequencies of aneuploidy in diploid androgenetic heterozygous and triploid diandric heterozygous conceptuses were significantly higher than the frequency of aneuploidy in diploid androgenetic homozygous conceptuses. In diploid androgenetic and triploid diandric conceptuses that are heterozygous for paternally inherited alleles, the two paternally inherited sets of genomes originate in two spermatozoa. Each spermatozoon provides one pair of centrioles to the zygote. The presence of two pairs of centrioles may cause an increased risk of aneuploidy.

Targeted next-generation sequencing of adult gliomas for retrospective prognostic evaluation and up-front diagnostics.

AIMS: We aimed to reclassify a population-based cohort of 529 adult glioma patients to evaluate the prognostic impact of the 2016 World Health Organization (WHO) central nervous system tumour classification. Moreover, we evaluated the feasibility of gene panel next-generation sequencing (NGS) in daily diagnostics of 225 prospective glioma patients. METHODS: The retrospective cohort was reclassified according to WHO 2016 criteria by immunohistochemistry for IDH-R132H, fluorescence in situ hybridization for 1p/19q-codeletion and gene panel NGS. All tumours of the prospective cohort were subjected to NGS analysis up-front. RESULTS: The entire population-based cohort was successfully reclassified according to WHO 2016 criteria. NGS results were obtained for 98% of the prospective patients. Survival analyses in the population-based cohort confirmed three major prognostic subgroups, that is, isocitrate dehydrogenase (IDH)-mutant and 1p/19q-codeleted oligodendrogliomas, IDH-mutant astrocytomas and IDH-wildtype glioblastomas. The distinction between WHO grade II and III was prognostic in patients with IDH-mutant astrocytoma. The survival of patients with IDH-wildtype diffuse astrocytomas carrying TERT promoter mutation and/or EGFR amplification overlapped with the poor survival of IDH-wildtype glioblastoma patients. CONCLUSIONS: Gene panel NGS proved feasible in daily diagnostics. In addition, our study confirms the prognostic role of glioma classification according to WHO 2016 in a large population-based cohort. Molecular features of glioblastoma in IDH-wildtype diffuse glioma were linked to poor survival corresponding to IDH-wildtype glioblastoma patients. The distinction between WHO grade II and III retained prognostic significance in patients with IDH-mutant diffuse astrocytic gliomas.

Detection of growth-restricted fetuses during pregnancy is associated with fewer intrauterine deaths but increased adverse childhood outcomes: an observational study.

OBJECTIVE: Exploring associations between antenatal detection of fetal growth restriction (FGR) and adverse outcome. DESIGN: Retrospective, observational, register-based study. SETTING: Zealand, Denmark. POPULATION OR SAMPLE: Children born from 1 September 2012 to 31 August 2015. METHODS: Diagnoses from birth until 1 January 2018 were retrieved from The National Patient Registry. Detection was defined as estimated fetal weight less than the 2.3rd centile. Cox regression was used to associate detection status with the hazard rate of adverse outcome, adjusted for fetal weight deviation, maternal age, ethnicity, body mass index and smoking. MAIN OUTCOME MEASURES: Adverse neonatal outcome, adverse neuropsychiatric outcome, respiratory disorders, endocrine disorders, gastrointestinal/urogenital disorders. RESULTS: A total of 2425 FGR children were included. An association was found for gastrointestinal/urogenital disorders (hazard ratio [HR] 1.68, 95% CI 1.26-2.23, P < 0.001) and respiratory disorders (HR 1.22, 95% CI 1.02-1.46, P = 0.03) in detected versus undetected infants. For adverse neuropsychiatric outcome, HR was 1.32 (95% CI 1.00-1.75, P = 0.05). There was no evidence of an association between detection and adverse neonatal outcome (HR 1.00, 95% CI 0.62-1.61, P = 0.99) and endocrine disorders (HR 1.39, 95% CI 0.88-2.19, P = 0.16). Detected infants were smaller (median -28% versus -25%, P < 0.0001), more often born preterm (odds ratio [OR] 4.15, 3.12-5.52, P < 0.0001) and more often born after induction or caesarean section (OR 5.19, 95% CI 4.13-6.51, P < 0.0001). Stillbirth risk was increased in undetected FGR fetuses (OR 2.63, 95% CI 1.37-5.04, P = 0.004). CONCLUSIONS: We found an association between detection of FGR and risk of adverse childhood conditions, possibly caused by prematurity. Iatrogenic prematurity may be inevitable in stillbirth prevention, but is accompanied by a risk of long-term childhood conditions. TWEETABLE ABSTRACT: Antenatal detection of growth-restricted fetuses is associated with adverse childhood outcomes but fewer intrauterine deaths.

Multicenter randomized trial exploring effects of simulation-based ultrasound training on obstetricians' diagnostic accuracy: value for experienced operators.

OBJECTIVE: To explore the effects of simulation-based ultrasound training on the accuracy of fetal weight estimation in the third trimester among obstetricians with different levels of clinical experience. METHODS: This was a multicenter, randomized pre-post-test practical trial conducted between March 2016 and January 2018. Obstetricians with different levels of clinical experience were randomized to either simulation-based ultrasound training focusing on fetal weight scans or no intervention. Participants completed two scans in pregnant women at term to establish baseline accuracy of fetal weight estimation. Another two scans were performed at follow-up. Accuracy was defined by the percentage difference between estimated fetal weight and actual birth weight. Ultrasound image quality was rated by two expert raters. RESULTS: Seventy participants with different levels of clinical experience completed the study. Adjusting for clinical experience, the intervention group demonstrated an improvement in measurement accuracy of 31.9% (95% CI, 6.9-50.1%) (P = 0.02), whereas the control group did not improve (relative difference, 13.1% (95% CI, -17.9 to 55.9%); P = 0.45). The change in accuracy was significantly different between the groups (P = 0.02) and independent of clinical experience (P = 0.54). Image-quality scores improved by a mean of 1.2 (95% CI, 0.4-2.1) (P < 0.01) in the intervention group, with no change in the control group (mean difference, 0.1 (95% CI, -0.8 to 1.0); P = 0.78). There was a strong negative correlation between time spent using the simulator and clinical experience (r = -0.70, P = 0.0001). CONCLUSION: Simulation-based ultrasound training improved accuracy and image quality when performing fetal weight estimation in women at term, independent of obstetricians' clinical experience. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd.

Simulation-based point-of-care ultrasound training: a matter of competency rather than volume.

BACKGROUND: Point-of-care ultrasonography plays an increasingly important role in the initial resuscitation of critically ill patients but acquisition of the skill is associated with long learning curves. The skills required to perform ultrasound examinations can be practiced in a simulated setting before being performed on actual patients. The aim of this study was to investigate the learning curves for novices training the FAST protocol on a virtual-reality simulator. METHODS: Ultrasound novices (N = 25) were instructed to complete a FAST training program on a virtual-reality ultrasound simulator. Participants were instructed to continue training until they reached a previously established mastery learning level, which corresponds to the performance level of a group of ultrasound experts. Performance scores and time used during each FAST examination were used to determine participants' learning curves. RESULTS: The participants attained the mastery learning level within a median of three (range two to four) attempts corresponding to a median of 1 h 46 min (range 1 h 2 min to 3 h 37 min) of simulation training. The ultrasound novices' examination speed improved significantly with training, and continued to improve even after they attained the mastery learning level (P = 0.011). Twenty-three participants attained the mastery learning level. CONCLUSION: Novices can attain mastery learning levels using simulation-based ultrasound training with less than, on average, 2 h of practice. However, we found large variations in the amount of training needed, which raises questions about the adequacy of current volume-based models for determining ultrasound competency.

The total pregnancy potential per oocyte aspiration after assisted reproduction-in how many cycles are biologically competent oocytes available?

PURPOSE: While stimulation of women prior to assisted reproduction is associated with increased success rates, the total biological pregnancy potential per stimulation cycle is rarely assessed. METHODS: Retrospective sequential cohort study of the cumulative live birth rate in 1148 first IVF/ICSI-cycles and 5-year follow up of frozen embryo replacement (FER) cycles were used. Oocyte number, number of embryos transferred, and cryopreserved/thawed and transferred embryos in a FER cycle were registered for all patients. Children per oocyte and per transferred embryo and percentage of cycles with births were calculated. RESULTS: We obtained 9529 oocytes. Embryos (2507) were transferred in either fresh or FER cycles, resulting in 422 births and 474 live born children. Median age of the women was 32.5 years (range 20-41.5 years). In total, 34.3 % of all cycles ended with a live birth while in 65.7 % of the cycles, no oocytes were capable of developing into a child. The average number of oocytes needed per live born child after transfer of fresh and thawed embryos was 20 as only 5.0 % of oocytes aspirated in the first IVF/ICSI cycle had the competence to develop into a child. CONCLUSIONS: In our setting, overall 5.0 % of the oocytes in a first cycle were biologically competent and in around 2/3 of all cycles, none of the oocytes had the potential to result in the birth of a child.

NLRP7 or KHDC3L genes and the etiology of molar pregnancies and recurrent miscarriage.

Women with mutation in both alleles of the NLRP7 or C6orf221/KHDC3L genes are predisposed to diploid biparental moles, but it has also been suggested that mutation in these genes can predispose to diploid androgenetic or triploid moles and to other kinds of reproductive wastage. We have investigated the association between molar pregnancy and recurrent miscarriages regarding changes in the NLRP7 and C6orf221/KHDC3L genes. Our study group can be divided into three sub-cohorts: (i) women having had at least one molar pregnancy and at least two non-mole miscarriages, (ii) women having had recurrent androgenetic hydatidiform moles and (iii) women having had one diploid androgenetic hydatidiform mole and a relative having had a hydatidiform mole (familial hydatidiform moles). We observed a statistically non-significant tendency of non-synonymous variants in NLRP7 to be more frequent in women with familial hydatidiform mole and in women with female family members with hydatidiform mole or non-mole miscarriage compared with women with no family history of mole or miscarriage. However, we did not find any unequivocal pathogenic mutations (the term 'unequivocal pathogenic mutations' refers to mutations that indubitably have a pathogenic effect on the affected woman) in NLRP7 or C6orf221/KHDC3L in any of the women in the study group. This indicates that recurrent miscarriages plus hydatidiform mole, recurrent androgenetic hydatidiform moles and familial androgenetic hydatidiform moles in general do not have the same monogenetic etiology as familiar diploid biparental moles.

Mosaic moles and non-familial biparental moles are not caused by mutations in NLRP7, NLRP2 or C6orf221.

Hydatidiform moles (HMs) most often occur sporadically and are either diploid androgenetic or triploid. The very rare familial recurrent HMs (FRHMs) have been related to NLRP7 and C6orf221 mutations in the mother. FRHMs are most often diploid with both maternal and paternal origin of the molar genome. We have screened a cohort of 11 women with diploid HMs with biparental contributions to the molar genome with regard to mutations in NLRP7, NLRP2, the NLRP gene most homologous to NLRP7, and C6orf221. This was done in order to reveal if mutations in the mentioned genes play a major role in development of non-recurrent biparental moles. Recently, we have shown that eight of these diploid moles consist of two different cell lines. Only one woman had a mutation in the coding DNA sequence of NLRP7, which most likely contributed to HM development. This woman had non-mosaic repeated moles, and she was the only woman in our cohort with FRHM. We found no unequivocal pathogenic mutations in NLRP2 or C6orf221. Our observations suggest that although NLRP7 and C6orf221 mutations are related to diploid biparental FRHMs, neither of these genes, nor NLRP2, are related to diploid HMs with biparental contributions to the molar genome, in general.

Measurement of the performance of nerve cuff electrodes for recording.

New designs of cuff electrodes for the recording of signals from peripheral nerves are typically tested in acute animal experiments before long-term evaluation takes place. A reproducible, cost-effective and fast method is presented for evaluating cuff electrodes with respect to signal amplitude, noise rejection, and, in some cases, selectivity, as an alternative to acute in vivo experiments. Comparisons with a computer model and with signals obtained from rabbit tibial nerve give good agreement with the new method. It is shown that an imperfect closure of the cuff around the nerve can easily lead to more than 50% loss of the signal amplitude. Noise from sources external to the cuff is not significantly affected by the closing mechanism, but is strongly reduced by a tripolar cuff configuration as compared with a monopolar one (reduction factor 2.8 to 58, mean = 6.5, n = 6). In dual-channel cuffs, cross-talk is below 1.2% indicating a very high selectivity.

[Screening against cervical cancer. Experiences, attitudes and knowledge of women in the county of Funen]. / Screening mod livmoderhalskraeft. Fynske kvinders erfaringer, holdninger og viden.

Questionnaires on women's attitudes and knowledge of cervical screening in the County of Funen were mailed to a sample of 1505 attenders aged 23-59 years, stratified on age and residence. A high proportion of attenders (80.4%) answered the questionnaire. Two-thirds of the women had been informed about the screening program before they received the invitation to participate. Generally the women were satisfied with the introduction to the screening program. However, the majority of the attenders were not satisfied with the way they received the result. They wanted the result no later than two weeks after the test was done, and they wanted the result from their GP. Independent of age and education the majority (about 90%) had a good general knowledge (knew that the smear is taken from the cervix; that early diagnosis is important; that cervix cancer can be treated). However the specific knowledge was lower and significantly associated with education (e.g. 43% versus 63% could identify the cervix on a drawing of the uterus among women with respectively low and high levels of school education). Furthermore, the majority were of the opinion that the test should be done more frequently than every three years as recommended by the National Health Board.

Causal probabilistic network and power spectral estimation used in sleep stage classification.

A new method for sleep-stage classification using a causal probabilistic network as automatic classifier has been implemented and validated. The system uses features from the primary sleep signals from the brain (EEG) and the eyes (AOG) as input. From the EEG, features are derived containing spectral information which is used to classify power in the classical spectral bands, sleep spindles and K-complexes. From AOG, information on rapid eye movements is derived. Features are extracted every 2 seconds. The CPN-based sleep classifier was implemented using the HUGIN system, an application tool to handle causal probabilistic networks. The results obtained using different training approaches show agreements ranging from 68.7 to 70.7% between the system and the two experts when a pooled agreement is computed over the six subjects. As a comparison, the interrater agreement between the two experts was found to be 71.4%, measured also over the six subjects.

Combined plasma-desorption mass spectrometry and Edman degradation applied to simultaneous sequence determination of isoforms of structural proteins from the cuticle of Locusta migratoria.

The primary structures of two basic low-molecular-mass proteins, Lm-67 and Lm-70 from the pharate cuticle of the migratory locust, Locusta migratoria, were determined. The sequencing strategy was based on combined use of plasma--desorption mass spectrometry (PDMS) and automatic Edman degradation of the proteins and their enzymically derived peptides. The mass-spectral data showed the presence of two proteins in each preparation. For protein preparation Lm-67, this was indicated by the mass spectrum of the intact protein. For protein preparation Lm-70, the presence of two variants only became evident by mass-spectrometric analysis of the enzymically derived peptides. Both proteins show strong similarity to other exocuticular proteins from L. migratoria.