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Rhabdomyosarcoma, although rare, is the most frequent soft tissue sarcoma in children and adolescents. It can present as a mass at nearly any site in the body, with most common presentations in the head and neck, genitourinary tract and extremities. The optimal diagnostic approach and management of rhabdomyosarcoma require a multidisciplinary team with multimodal treatment, including chemotherapy and local therapy. Survival has improved over the last decades; however, further improvement in management is essential with current 5-year overall survival ranging from 35% to 100%, depending on disease and patient characteristics. In the full patient journey, from diagnosis, staging, management to follow-up after therapy, the paediatric radiologist and nuclear physician are essential members of the multidisciplinary team. Recently, guidelines of the European paediatric Soft tissue sarcoma Study Group, the Cooperative Weichteilsarkom Studiengruppe and the Oncology Task Force of the European Society of Paediatric Radiology (ESPR), in an ongoing collaboration with the International Soft-Tissue Sarcoma Database Consortium, provided guidance for high-quality imaging. In this educational paper, given as a lecture during the 2022 postgraduate ESPR course, the multi-disciplinary team of our national paediatric oncology centre presents the journey of two patients with rhabdomyosarcoma and discusses the impact on and considerations for the clinical (paediatric) radiologist and nuclear physician. The key learning points of the guidelines and their implementation in clinical practice are highlighted and up-to-date insights provided for all aspects from clinical suspicion of rhabdomyosarcoma and its differential diagnosis, to biopsy, staging, risk stratification, treatment response assessment and follow-up.

RNA sequencing is a powerful tool to analyze gene expression transcriptome wide. However, RNA sequencing in general and especially the recently developed methods of long read RNA sequencing are still low-throughput and cost-intensive. Here, one important design choice is to concentrate the sequencing capacity on specific parts of the transcriptome. Especially, abundant transcripts as ribosomal RNAs may dominate the available sequencing space, if not removed prior to sequencing. Several methods exist to reduce ribosomal RNA read numbers: either based on enrichment of the relevant fraction (polyA+ RNA) or depletion, respectively degradation of ribosomal RNAs. Furthermore, commercial kits are available to deplete globin transcripts from blood samples. However, so far, no solution exists to deal with other tissue-specific highly abundant transcripts. This is especially of interest in the heart and other muscle derived samples, where reads originating from mitochondrial RNAs make up to 30% of reads in polyA+ selected libraries and around 70% in single cell sequencing experiments. We present a simple method to diminish sequencing of mitochondrial RNAs in Oxford Nanopore direct RNA sequencing libraries by RNase H based clipping of the polyA tail. We show that mt-clipping enables enhanced detection of cytoplasmic mRNAs, among them genes involved in heart development and pathogenesis. Mt-clipping may be applied as well to other sequencing protocols that are based on oligo(dT) priming and can be easily adapted to other tissue-specific high-abundant transcripts.

We introduce Single-cell Nanopore Spatial Transcriptomics (scNaST), a software suite to facilitate the analysis of spatial gene expression from second- and third-generation sequencing, allowing to generate a full-length near-single-cell transcriptional landscape of the tissue microenvironment. Taking advantage of the Visium Spatial platform, we adapted a strategy recently developed to assign barcodes to long-read single-cell sequencing data for spatial capture technology. Here, we demonstrate our workflow using four short axis sections of the mouse heart following myocardial infarction. We constructed a de novo transcriptome using long-read data, and successfully assigned 19,794 transcript isoforms in total, including clinically-relevant, but yet uncharacterized modes of transcription, such as intron retention or antisense overlapping transcription. We showed a higher transcriptome complexity in the healthy regions, and identified intron retention as a mode of transcription associated with the infarct area. Our data revealed a clear regional isoform switching among differentially used transcripts for genes involved in cardiac muscle contraction and tissue morphogenesis. Molecular signatures involved in cardiac remodeling integrated with morphological context may support the development of new therapeutics towards the treatment of heart failure and the reduction of cardiac complications.

The catalytically active component of ribosomes, rRNA, is long studied and heavily modified. However, little is known about functional and pathological consequences of changes in human rRNA modification status. Direct RNA-sequencing on the Nanopore platform enables the direct assessment of rRNA modifications. We established a targeted Nanopore direct rRNA sequencing approach and applied it to CRISPR-Cas9 engineered HCT116 cells, lacking specific enzymatic activities required to establish defined rRNA base modifications. We analyzed these sequencing data along with wild type samples and in vitro transcribed reference sequences to specifically detect changes in modification status. We show for the first time that direct RNA-sequencing is feasible on smaller, i.e. Flongle, flow cells. Our targeted approach reduces RNA input requirements, making it accessible to the analysis of limited samples such as patient derived material. The analysis of rRNA modifications during cardiomyocyte differentiation of human induced pluripotent stem cells, and of heart biopsies from cardiomyopathy patients revealed altered modifications of specific sites, among them pseudouridines, 2’-O-methylation of riboses and an acetylation of cytidine that was validated by orthogonal methods and the use of a genetic model. Targeted direct rRNA-sequencing analysis with JACUSA2 opens up the possibility to analyze dynamic changes in rRNA modifications in a wide range of biological and clinical samples.

Repetitive reorganizations cause change fatigue, which creates employee resistance to further organizational change. This paper investigates how such fatigue arises. The results show that uncertain...

ONT long-read sequencing provides real-time monitoring and controlling of individual nanopores. Adaptive sampling enriches or depletes specific sequences in Nanopore DNA sequencing, but was not applicable to direct sequencing of RNA so far.Here, we identify essential parameter settings for direct RNA sequencing (DRS). We demonstrate the superior performance of depletion over enrichment and show that adaptive sampling efficiently depletes specific transcripts in transcriptome-wide sequencing applications. Specifically, we applied our adaptive sampling approach to polyA+ RNA samples from human cardiomyocytes and mouse whole heart tissue. Herein, we show more than 2.5-fold depletion of highly abundant mitochondrial-encoded transcripts that in normal sequencing account for up to 40% of sequenced bases in heart tissue samples.

The coronavirus disease 2019 (COVID-19) pandemic has led to preventive measures worldwide. With the decline of infection rates, less stringent restrictions for sports and exercise are being implemented. COVID-19 is associated with significant cardiovascular complications; however there are limited data on cardiovascular complications and long-term outcomes in both competitive (elite) athletes and highly active individuals. Based on different categories of disease severity (asymptomatic, regional/systemic symptoms, hospitalisation, myocardial damage, and/or myocarditis), in this point-of-view article we offer the (sports) cardiologist or sports physician in the Netherlands a practical guide to pre-participation screening, and diagnostic and management strategies in all athletes >16 years of age after COVID-19 infection.

There is broad consensus on the utility of complex pictures in the assessment of simultanagnosia in patients with acquired brain injury (ABI). To overcome various shortcomings of current instruments, we have developed the Birthday Party Test (BPT); a picture description test that contains a neutral scene, a balanced representation of events, and provides clear instructions and a scoring-aid. We have applied the BPT in a large group of patients with ABI (n = 502) and in an age-matched healthy control group (n = 194). Our results show that performance on the BPT was associated with a range of descriptive, neuropsychological and clinical characteristics and that poor test performance appeared to be more common in patients with etiologies that have an increased risk of bilateral damage. Furthermore, we assume a high correspondence between test performance on the BPT and the assessor’s clinical judgment of likely having simultanagnosia in preliminary analyses. This study shows the potential usefulness of the BPT to support diagnostic decision making in simultanagnosia. The BPT is made freely available to facilitate its broad application in the clinical assessment of patients with visual impairment and to enable a further evaluation of its utility and validity in future studies.

Briefings in bioinformatics 22(6), 6 (2021). doi:10.1093/bib/bbab219, Published by Oxford University Press, Oxford [u.a.]

To conserve water in arid environments, numerous plant lineages have independently evolved Crassulacean Acid Metabolism (CAM). Interestingly, Isoetes, an aquatic lycophyte, can also perform CAM as an adaptation to low CO2 availability underwater. However, little is known about the evolution of CAM in aquatic plants and the lack of genomic data has hindered comparison between aquatic and terrestrial CAM. Here, we investigated the underwater CAM in Isoetes taiwanensis by generating a high-quality genome assembly and RNA-seq time course. Despite broad similarities between CAM in Isoetes and terrestrial angiosperms, we identified several key differences. Notably, for carboxylation of PEP, Isoetes recruited the lesser-known “bacterial-type” PEPC, along with the “plant-type” exclusively used in other terrestrial CAM and C4 plants. Furthermore, we found that circadian control of key CAM pathway genes has diverged considerably in Isoetes relative to flowering plants. This suggests the existence of more evolutionary paths to CAM than previously recognized.

AimsWe aimed to assess and characterize sex differences in adverse drug reactions (ADRs) reported to the national pharmacovigilance centre in the Netherlands while considering differences in drug use.MethodsADRs spontaneously reported by healthcare professionals and patients to the Netherlands pharmacovigilance centre Lareb were used. Drug-ADR combinations reported at least 10 times between 2003-2016 for drugs used by 10,000 persons in that period were included. Data about the number of drug users was obtained from the National Health Care Institute. Sex-specific ADRs, like gynaecological problems, were excluded. Sex differences in specific drug-ADR combinations were tested using bivariate logistic regression analyses in which the number of drug users per sex was taken into account.ResultsIn total, 2483 drug-ADR combinations were analysed. Possibly relevant sex differences were shown in 363 combinations (15%). Most of these drug-ADR combinations were reported more often for women (322 combinations). Drugs with the highest number of ADRs that were more often reported for women included thyroid hormones (32 combinations) and antidepressants (16 combinations for the centrally acting sympathomimetics; 14 combinations for other antidepressants). Some ADRs were predominantly reported for women across a range of drugs such as headache and dizziness whereas other ADRs such as tendon ruptures and aggression were reported more often for men.ConclusionsIdentified sex differences in reported ADRs often referred to women. These differences may have various causes, including pharmacological and behavioural causes, which need to be further assessed. The results may ultimately lead to sex-specific prescribing or monitoring recommendations.

Acute myeloid leukemia is an aggressive disease that arises from clonal expansion of malignant hematopoietic precursor cells of the bone marrow. Deletions on the long arm of chromosome 9 (del(9q)) are observed in 2% of acute myeloid leukemia patients. Our deletion analysis in a cohort of 31 del(9q) acute myeloid leukemia patients further supports the importance of a minimally deleted region composed of seven genes potentially involved in leukemogenesis: GKAP1, KIF27, C9ORF64, HNRNPK, RMI1, SLC28A3 and NTRK2. Importantly, among them HNRNPK, encoding heterogeneous nuclear ribonucleoprotein K is proposed to function in leukemogenesis. We show that expression of HNRNPK and the other genes of the minimally deleted region is significantly reduced in patients with del(9q) compared with normal karyotype acute myeloid leukemia. Also, two mRNAs interacting with heterogeneous nuclear ribonucleoprotein K, namely CDKN1A and CEBPA are significantly downregulated. While the deletion size is not correlated with outcome, associated genetic aberrations are important. Patients with an additional t(8;21) show a good prognosis. RUNX1-RUNX1T1, which emerges from the t(8;21) leads to transcriptional down-regulation of CEBPA. Acute myeloid leukemia patients with mutations in CEBPA have a good prognosis as well. Interestingly, in del(9q) patients with CEBPA mutation mRNA levels of HNRNPK and the other genes located in the minimally deleted region is restored to normal karyotype level. Our data indicate that a link between CEBPA and the genes of the minimally deleted region, among them HNRNPK contributes to leukemogenesis in acute myeloid leukemia with del(9q).

The main aim of this study was to determine the agreement in classification between the modified KörperKoordinations Test für Kinder (KTK3+) and the Athletic Skills Track (AST) for measuring fundamental movement skill levels (FMS) in 6- to 12-year old children. 3,107 Dutch children (of which 1,625 are girls) between 6 and 12 years of age (9.1 ± 1.8 years) were tested with the KTK3+ and the AST. The KTK3+ consists of three items from the KTK and the Faber hand-eye coordination test. Raw scores from each subtest were transformed into percentile scores based on all the data of each grade. The AST is an obstacle course consisting of 5 (grades 3 till 5, 6–9 years) or 7 (grades 6 till 8, 9–12 years) concatenated FMS that should be performed as quickly as possible. The outcome measure is the time needed to complete the track. A significant bivariate Pearson correlation coefficient of 0.51 was found between the percentile sum score of the KTK3+ and the time to complete the AST, indicating that both tests measure a similar construct to some extent. Based on their scores, children were classified into one of five categories: 95%. Cross tabs revealed an agreement of 58.8% with a Kappa value of 0.15 between both tests. Less than 1% of the children were classified more than two categories higher or lower. The moderate correlation between the KTK3+ and the AST and the low classification agreement into five categories of FMS stress the importance to further investigate the test choice and the measurement properties (i.e., validity and reliability) of both tools. PE teachers needs to be aware of the context in which the test will be conducted, know which construct of motor competence they want to measure and know what the purpose of testing is (e.g., screening or monitoring). Based on these considerations, the most appropriate assessment tool can be chosen.

The Implanon NXT is a commonly used contraceptive. Incorrect localization of the implant can cause complications.A 41-year-old woman is seen in the gynaecology outpatient clinic with a request to remove a recently placed Implanon NXT because of worsening mood symptoms. The implant can't be found on physical and ultrasound examination. Duringsurgicalexplorationthe implant is not found at theinsertion site' By means of X-ray scanning the implant becomes visible around the humeral head. The implant appears to be located in the cephalic vein and is subsequently removed.In case of a referral due to because of worsening mood symptoms after an Implanon NXT exchange, it is possible that the implant is localized incorrectly. It is recommended to use additional imaging before performing surgical exploration. Furthermore, it is important to insert the Implanon NXT according to the supplied instructions to prevent this complication.

Sports cardiology is a rapidly evolving subspecialty of cardiology, with a growing demand for expertise. To improve patient care, clinicians, patients, and athletes (recreational to elite) should be able to easily identify specialised care pathways, expertise centres and clinicians with sports cardiology expertise. To this purpose, several international societies and organisations recommend establishing a local and national sports cardiology infrastructure. We therefore aimed to establish The Netherlands Sports Cardiology Map. We conducted a web-based survey, which was published on the Netherlands Society of Cardiology home page (2019–2020) and in which each cardiology department or clinic was asked to provide information on sports cardiology expertise and the current infrastructure. Of the 46 respondent centres, 28 (61%) reported that they had expertise in sports cardiology, of which 22 (79%) had specific expertise in one or more specific types of sports. Integrated multidisciplinary meetings were reported by 43% of the centres (n = 12/28). Only two centres reported ongoing research projects that had been approved by an institutional review board. The Netherlands Sports Cardiology Map is an important step towards improving the existing infrastructure and developing network medicine for sports cardiology.

The complement system is implicated in synapse loss in the MS hippocampus, but the functional consequences of synapse loss remain poorly understood. Here, in post-mortem MS hippocampi with demyelination we find that deposits of the complement component C1q are enriched in the CA2 subfield, are linked to loss of inhibitory synapses and are significantly higher in MS patients with cognitive impairments compared to those with preserved cognitive functions. Using the cuprizone mouse model of demyelination, we corroborated that C1q deposits are highest within the demyelinated dorsal hippocampal CA2 pyramidal layer, and co-localized with inhibitory synapses engulfed by microglia/macrophages. In agreement with the loss of inhibitory perisomatic synapses, we further found that Schaffer collateral feedforward inhibition but not excitation was impaired in CA2 pyramidal neurons and accompanied by a reduced spike output. Ultimately, we show that these electrophysiological changes were associated with an impaired encoding of social memories. Together, our findings identify CA2 as a critical circuit in demyelinated intrahippocampal lesions and memory dysfunctions in MS.

Nucleic acids Research 49(6), 3507-3523 (2021). doi:10.1093/nar/gkab116, Published by Oxford Univ. Press, Oxford

Frontiers in molecular biosciences 8, 625608 (2021). doi:10.3389/fmolb.2021.625608, Published by Frontiers, Lausanne

Impairments in visual perception are common after acquired brain injury (ABI) and adequate assessment is crucial for diagnosis and rehabilitation. However, there is no consensus yet on how to assess these disorders after ABI. The aim of the present study was to explore what measures are considered reasonable to be part of a test battery for the screening of a broad range of mid-level and higher-order visual perceptual disorders. A Delphi method was used to collect the opinions of 28 international multidisciplinary experts in visual perception in order to achieve consensus on the content of the test battery. Seventeen experts evaluated the test battery proposed in the third and final round of the Delphi process. Consensus was achieved (94%) on a battery of 11 distinctive tests with an expected administration time of 30 minutes. The current study provides an essential step in the development of a standardized and time-efficient test battery for the screening of mid-level and higher-order visual perceptual disorders. The composed battery may improve effectiveness of clinical assessment by providing insight into potential visual deficits in little time, thereby initiating further assessment. Future studies should focus on the validation of the suggested test battery and collect normative data.

Massively parallel RNA sequencing (RNA-seq) in combination with metabolic labeling has become the de facto standard approach to study alterations in RNA transcription, processing or decay. Regardless of advances in the experimental protocols and techniques, every experimentalist needs to specify the key aspects of experimental design: For example, which protocol should be used (biochemical separation vs. nucleotide conversion) and what is the optimal labeling time? In this work, we provide approximate answers to these questions using the asymptotic theory of optimal design. Specifically, we investigate, how the variance of degradation rate estimates depends on the time and derive the optimal time for any given degradation rate. Subsequently, we show that an increase in sample numbers should be preferred over an increase in sequencing depth. Lastly, we provide some guidance on use cases when laborious biochemical separation outcompetes recent nucleotide conversion based methods (such as SLAMseq) and show, how inefficient conversion influences the precision of estimates. Code and documentation can be found at https://github.com/dieterich-lab/DesignMetabolicRNAlabeling., Author summary Massively parallel RNA sequencing (RNA-seq) in combination with metabolic labeling has become the de facto standard approach to study alterations in RNA transcription, processing or decay. In our manuscript, we address several key aspects of experimental design: 1) The optimal labeling time, 2) the number of replicate samples over sequencing depth and 3) the choice of experimental protocol. We provide approximate answers to these questions using asymptotic theory of optimal design.

In a cyclical Fitts' task, hand movements transition from continuous to discrete movements when the Index of Difficulty (ID) increases. Moreover, at high ID (small target), the eyes saccade to and subsequently fixate the targets at every movement, while at low ID (large target) intermittent monitoring is used. By hypothesis, the (periodic) gaze shifts are abandoned for movement times shorter than about 0.350s due to systemic constraints (i.e., a refractory period and intrinsic latency). If so, the transition in eye and hand movements is independent. To investigate these issues, the present study examined the effects of changing ID via the targets' width or distance as well as hysteresis in eye-hand coordination. To this aim, 14 participants performed a cyclical Fitts' task while their hand and eye movements were recorded simultaneously. The results show that the transition in eye-hand synchronization (at 2.87 bit; 0.25s) and in hand dynamics (at 4.85 bit; 0.81s) neither co-occurred nor correlated. Some small width vs. distance dissociations and hysteresis effects were found, but they disappeared when eye-hand synchronization was viewed as a function of movement time rather than ID. This confirms that a minimal between-saccade time is the limiting factor in eye-hand synchronization. Additionally, the timing between the start of the hand movement and the saccade appeared to be relatively constant (at 0.15s) and independent of movement time, implying a constant delay that should be implemented in a dynamical model of eye-hand coordination.

Background Aim of the study was to test the 12-month effects of a multicomponent physical activity (PA) intervention at schoolyards on recess PA levels of 6th and 7th grade children in primary schools, combining Global Positioning System (GPS) and accelerometry data. Methods A quasi-experimental study design was used with 20 paired intervention and control schools. GPS confirmatory analyses were applied to validate attendance at schoolyards during recess. Data from 376 children from 7 pairs of schools were included in the final analyses. Pooled intervention effectiveness was tested by multilevel linear regression analyses, whereas effectiveness of intervention components was tested by multivariate linear regression analyses. Results Children exposed to the multicomponent intervention increased their time spent in light PA (+5.9%) during recess. No pooled effects on Moderate-to-Vigorous PA were found. In depth-analyses on intervention components showed that physical schoolyard interventions particularly predicted a decrease in time spent in sedentary behavior during recess at follow-up. Intervention intensity and school's commitment to the project strengthened this effect. Conclusions The multicomponent schoolyard PA intervention was effective in making children spend a larger proportion of recess time in light PA, which was most likely the result of a shift from sedentary behavior to light PA.

Item does not contain fulltext People who grow up under stressful conditions tend to score lower on conventional assessments of cognitive abilities. They might, however, develop enhanced mental skills and abilities for solving problems that are more ecologically relevant to them. We present 2 studies examining whether psychosocial adversity (i.e., exposure to neighborhood violence and to harsh parenting) enhances the ability to detect deception. Our results, based on Signal Detection and Bayesian analyses, are mixed. In Study 1, we find no support for our hypothesis that exposure to psychosocial adversity enhances deception detection ability. In Study 2, we do find that only harsher parenting predicts greater accuracy. This evidence is either weak or strong, depending on whether we compare our hypothesis to the null hypothesis or an impairment model, respectively. In both studies, we find no relationship between neighborhood violence and accuracy. Although the implications of our findings are modest, we hope they will encourage future research focused on the skills and abilities of people who develop in harsh environments. 12 p.

Post-transcriptional regulation is crucial for structural and functional alterations in erythropoiesis. Enucleation of erythroid progenitors precedes reticulocyte release into circulation. In enucleated cells, reticulocyte 15-lipoxygenase (r15-LOX, also known as ALOX15) initiates mitochondria degradation. Regulation of r15-LOX mRNA translation by hnRNP K determines timely r15-LOX synthesis in terminal maturation. K562 cells induced for erythroid maturation recapitulate enucleation and mitochondria degradation. HnRNP K depletion from maturing K562 cells results in enhanced enucleation, which even occurs independently of maturation. We performed RIP-Chip analysis to identify hnRNP K-interacting RNAs comprehensively. Non-muscle myosin heavy chain (NMHC) IIA (also known as MYH9) mRNA co-purified with hnRNP K from non-induced K562 cells, but not from mature cells. NMHC IIA protein increase in erythroid maturation at constant NMHC IIA mRNA levels indicates post-transcriptional regulation. We demonstrate that binding of hnRNP K KH domain 3 to a specific sequence element in the NMHC IIA mRNA 3'UTR mediates translation regulation in vitro Importantly, elevated NMHC IIA expression results in erythroid-maturation-independent enucleation as shown for hnRNP K depletion. Our data provide evidence that hnRNP-K-mediated regulation of NMHC IIA mRNA translation contributes to the control of enucleation in erythropoiesis.

Introduction: Acute myeloid leukemia (AML) is characterized by heterogeneous cytogenetic and molecular aberrations. Deletions on the long arm of chromosome 9 (del(9q)) are observed in 2% of AML patients. In about 24% of the cases, del(9q) is observed as sole karyotypic abnormality, while in the remaining 76%, it is associated with a t(8;21) translocation or other aberrations. Among all del(9q) AML cases, 36%-50% exhibit an additional t(8;21), whereas 7%-14% of AML cases with t(8;21) show del(9q) as an additional aberration. A commonly deleted region (CDR) of del(9q) was defined and further analysis specified a minimally deleted region (MDR) composed of seven annotated genes (GKAP1, KIF27, C9ORF64, HNRNPK, RMI1, SLC28A3 and NTRK2) (Kronke J et al. Blood. 2013). However, the function of these genes and their impact on the pathogenesis of AML remain elusive. A recent study demonstrated that reduced expression of the HNRNPK gene product can contribute to leukemogenesis in AML (Gallardo M, Cancer Cell. 2015). The multifunctional protein hnRNP K interacts with other proteins, DNA and RNA, to modulate gene activity and gene expression on different levels. For example, hnRNP K not only regulates SRC gene transcription, but as well SRC mRNA translation and the activity of c-Src kinase. In the context of AML, hnRNP K was shown to interact with the mRNAs encoding C/EBPa (CEBPA) and p21 (CDKN1A). We analyzed a cohort of 31 del(9q) AML patients in order to further analyze the deleted region and to analyze the impact of HNRNPK deletion on leukemogenesis. Methods: 31 del(9q) patients were used for the characterization of the deleted region. mRNA level (determined by RT-qPCR analysis) and clinical parameters were compared with a cohort of 24 normal karyotype (NK) AML patients. HnRNP K immunoprecipitation was combined with RNA-Seq, a whole transcriptome shotgun sequencing application based on next generation sequencing and validated by RT-qPCR analysis. CRISPR-Cas9 genome editing has been applied to functionally characterize the impact of post-transcriptional control by hnRNP K in pathogenesis of AML. Results: Our analysis confirmed the MDR in a cohort of 31 AML del(9q) patients. Survival of patients and clinical parameters were not correlated with deletion size, further supporting the importance of the MDR, while other deleted genes seem to be less important for leukemogenesis. As demonstrated by qPCR analysis, the mRNA level of HNRNPK and other genes located in the MDR was reduced in patients carrying a del(9q) compared to NK patients. To further dissect a potential function of hnRNP K in AML del(9q), we characterized hnRNP K interacting mRNAs in the AML cell line KG-1a. Therefore, hnRNP K was immunoprecipitated from cytoplasmic extracts of KG-1a cells and interacting RNAs were identified by RNA-Seq analysis. This analysis revealed that 1076 RNAs are potentially associated with hnRNP K, among them the C/EBPa mRNA. Panther Protein Class analysis identified a high number of transcripts encoding nucleic acid binding proteins, mainly transcription factors. KG-1a cell lines harboring either a complete knock out of hnRNP K or a deletion of the RNA-binding KH-domain are currently generated by CRISPR-Cas9 genome editing to functionally analyze the impact of hnRNP K-mediated post-transcriptional control in AML. Conclusion and Outlook: The deletion of seven genes (GKAP1, KIF27, C9ORF64, HNRNPK, RMI1, SLC28A3 and NTRK2) in the MDR is indispensable, indicating a crucial function for the development of AML del(9q). Among them HNRNPK seems to be a particularly important factor in this process. The identification of hnRNP K interacting RNAs provides the basis to further improve our insight in molecular mechanisms, which drive the pathogenesis of AML del(9q). HNRNPK knock out cell lines will be used to analyze the effect of HNRNPK deletion on post-transcriptional control of identified target genes. Disclosures Ehninger: Cellex Gesellschaft fuer Zellgewinnung mbH: Employment, Equity Ownership; GEMoaB Monoclonals GmbH: Employment, Equity Ownership; Bayer: Research Funding. Brümmendorf:Merck: Consultancy; Janssen: Consultancy; Novartis: Consultancy, Research Funding; Pfizer: Consultancy, Research Funding; Takeda: Consultancy. Rollig:Janssen: Research Funding; Bayer: Research Funding. Thiede:Novartis: Honoraria, Research Funding; AgenDix: Other: Ownership.

Black and minority ethnic (BME) populations are disproportionately detained in psychiatric hospitals. To examine the dangerousness criteria for compulsory court ordered admission to a psychiatric hospital in White and BME persons. We examined the psychiatric examinations for court ordered compulsory admissions in 506 White and 299 BME persons from October 2004 until January 2008 in Rotterdam, the Netherlands. The White and BME groups are compared using Chi-square tests and in case of significant differences with logistic regression models adjusted for age, gender, mental disorders and socio-economic background. In BME persons, violence towards others and neglect of relatives were more often reasons to request court order admission as compared with Whites (39.8 vs. 25.3%, P < 0.001, respectively, 6.4 vs. 2.4%, P = 0.01). This remained true after adjustment for age, gender, mental disorders and socio-economic background [OR 1.56 (95% CI 1.12-2.18), P = 0.01, respectively; OR 3.08 (95% CI 1.31-7.26), P = 0.01]. The other reasons for a request of court order admission had a similar prevalence in both groups (suicide or self-harm, social decline, severe self-neglect, arousal of aggression of others, danger to the mental health of others, and the general safety of persons and goods). Violence towards others and neglect of relatives are more often a reason to request court ordered admission in BME than in White persons. BME patients are more often perceived as potentially dangerous to others.

Aim: Because the Dutch population has a growing number of older people, an increasing burden on mental health services is expected. To facilitate policy making for the future, it is important to know what changes there have been in use of mental health services by elderly in the past. This study investigates changes in the use of mental health services by older adults in the period 1990-2004. Methods: Information about the use of mental health services by older adults was retrieved from the Dutch Psychiatric Case Registers. Population size in these register areas and the unit costs of the different mental health services were taken into account. Results: In total there was an increase in the number of older adults that used mental health services in the period mentioned above. The costs, however, showed a decrease, which was caused by the decrease of expensive inpatient care and the increase of less expensive outpatient care. This was mainly the case until 2002. From this year on the ratio between inpatient and outpatient care stabilized. Conclusion: Deinstitutionalization of mental health care for older adults was shown in the period 1990-2002. This means that expensive inpatient care is partly replaced by less expensive outpatient care. As a consequence more older adults can be treated with no rise in costs. Since 2002 deinstitutionalization came to a halt. Because a growing number of older adults will be using mental health services in the future, new forms of outpatient care should be explored.

Post-transcriptional regulation is crucial for structural and functional alterations in erythropoiesis. Enucleation of erythroid progenitors precedes reticulocyte release into circulation. In enucleated cells, reticulocyte 15-lipoxygenase (r15-LOX, also known as ALOX15) initiates mitochondria degradation. Regulation of r15-LOX mRNA translation by hnRNP K determines timely r15-LOX synthesis in terminal maturation. K562 cells induced for erythroid maturation recapitulate enucleation and mitochondria degradation. HnRNP K depletion from maturing K562 cells results in enhanced enucleation, which even occurs independently of maturation. We performed RIP-Chip analysis to identify hnRNP K-interacting RNAs comprehensively. Non-muscle myosin heavy chain (NMHC) IIA (also known as MYH9) mRNA co-purified with hnRNP K from non-induced K562 cells, but not from mature cells. NMHC IIA protein increase in erythroid maturation at constant NMHC IIA mRNA levels indicates post-transcriptional regulation. We demonstrate that binding of hnRNP K KH domain 3 to a specific sequence element in the NMHC IIA mRNA 3'UTR mediates translation regulation in vitro Importantly, elevated NMHC IIA expression results in erythroid-maturation-independent enucleation as shown for hnRNP K depletion. Our data provide evidence that hnRNP-K-mediated regulation of NMHC IIA mRNA translation contributes to the control of enucleation in erythropoiesis.

Nucleic acids symposium series 43(6), 3219-3236 (2015). doi:10.1093/nar/gkv167, Published by Oxford Univ. Press, Oxford

Analysis of arginine methylation, which affects specific protein interactions in eukaryotic cells, requires access to methylated protein for biophysical and biochemical studies. Methylation of heterogeneous nuclear ribonucleoprotein K (hnRNP K) upon co-expression with protein arginine methyltransferase 1 in E. coli was monitored by mass spectrometry and found to be identical to the modification of hnRNP K purified from mammalian cells. Recombinant non-methylated and arginine-methylated hnRNP K (MethnRNP K) were used to characterize self-aggregation and nucleic acid binding. Analytical ultracentrifugation and static light scattering experiments revealed that hnRNP K methylation does not impact reversible self-aggregation, which can be prevented by high ionic strength and organic additives. Filter binding assays were used to compare the binding of non-methylated and MethnRNP K to the pyrimidine repeat-containing differentiation control element (DICE) of reticulocyte 15-lipoxygenase mRNA 3′ UTR. No affinity differences were detected for both hnRNP K variants. A series of oligonucleotides carrying various numbers of C4 motifs at different positions was used in steady state competition assays with fluorescently-labeled functional differentiation control element (2R). Quantitative evaluation indicated that all hnRNP K homology domains of hnRNP K contribute differentially to RNA binding, with KH1–KH2 acting as a tandem domain and KH3 as an individual binding domain.

We have measured the photon number probability distribution of a laser in which the inversion is not slaved to the field. For the experiments, we have used a Nd(3+):YVO(4) laser which has a sufficiently slow inversion to allow measurement of the photon fluctuations at a time scale much shorter than that of the relaxation oscillations. The photon distribution function becomes highly nonstandard (i.e., non-Poissonian) in such a laser; this is consistent with available theoretical work. We point out the relevance of our results for the case of the semiconductor microlaser.

We study the subthreshold growth of cracks in specimens with central crack made of AA5083-H321 aluminum alloy in air and seawater for various positive values of the load ratio. To this end, we use two different methods. In the first case, according to the ASTM E-647 standard, we gradually decrease the value of ΔK for constant R. In the second case, we gradually decrease Kmin for constant Kmax and, hence, increasing R. The results of both studies are in relatively good agreement. The insignificant difference between the thresholds ΔKth in air and seawater is, most likely, explained by the deposits of corrosion products.