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Background: Home treatment in child and adolescent psychiatry offers an alternative to conventional inpatient treatment by involving the patient's family, school, and peers more directly in therapy. Although several reviews have summarised existing home treatment programmes, evidence of their effectiveness remains limited and data synthesis is lacking., Methods: We conducted a meta-analysis on the effectiveness of home treatment compared with inpatient treatment in child and adolescent psychiatry, based on a systematic search of four databases (PubMed, CINAHL, PsychINFO, Embase). Primary outcomes were psychosocial functioning and psychopathology. Additional outcomes included treatment satisfaction, duration, costs, and readmission rates. Group differences were expressed as standardised mean differences (SMD) in change scores. We used three-level random-effects meta-analysis and meta-regression and conducted both superiority and non-inferiority testing., Results: We included 30 studies from 13 non-overlapping samples, providing data from 1795 individuals (mean age: 11.95 ± 2.33 years; 42.5% female). We found no significant differences between home and inpatient treatment for postline psychosocial functioning (SMD = 0.05 [- 0.18; 0.30], p = 0.68, I 2  = 98.0%) and psychopathology (SMD = 0.10 [- 0.17; 0.37], p = 0.44, I 2  = 98.3%). Similar results were observed from follow-up data and non-inferiority testing. Meta-regression showed better outcomes for patient groups with higher levels of psychopathology at baseline and favoured home treatment over inpatient treatment when only randomised controlled trials were considered., Conclusions: This meta-analysis found no evidence that home treatment is less effective than conventional inpatient treatment, highlighting its potential as an effective alternative in child and adolescent psychiatry. The generalisability of these findings is reduced by limitations in the existing literature, and further research is needed to better understand which patients benefit most from home treatment., Trial Registration: Registered at PROSPERO (CRD42020177558), July 5, 2020., (© 2024. The Author(s).)

Delayed fatherhood results in a higher risk of inheriting a new germline mutation that might result in a congenital disorder in the offspring. In particular, some FGFR3 mutations increase in frequency with age, but there are still a large number of uncharacterized FGFR3 mutations that could be expanding in the male germline with potentially early- or late-onset effects in the offspring. Here, we used digital polymerase chain reaction to assess the frequency and spatial distribution of 10 different FGFR3 missense substitutions in the sexually mature male germline. Our functional assessment of the receptor signaling of the variants with biophysical methods showed that 9 of these variants resulted in a higher activation of the receptor´s downstream signaling, resulting in 2 different expansion behaviors. Variants that form larger subclonal expansions in a dissected postmortem testis also showed a positive correlation of the substitution frequency with the sperm donor's age, and a high and ligand-independent FGFR3 activation. In contrast, variants that measured high FGFR3 signaling and elevated substitution frequencies independent of the donor's age did not result in measurable subclonal expansions in the testis. This suggests that promiscuous signal activation might also result in an accumulation of mutations before the sexual maturation of the male gonad with clones staying relatively constant in size throughout time. Collectively, these results provide novel insights into our understanding of the mutagenesis of driver mutations and their resulting mosaicism in the male germline with important consequences for the transmission and recurrence of associated disorders., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

Advanced paternal age increases the risk of transmitting de novo germline mutations, particularly missense mutations activating the receptor tyrosine kinase (RTK) signalling pathway, as exemplified by the FGFR3 mutation, which is linked to achondroplasia (ACH). This risk is attributed to the expansion of spermatogonial stem cells carrying the mutation, forming sub-clonal clusters in the ageing testis, thereby increasing the frequency of mutant sperm and the number of affected offspring from older fathers. While prior studies proposed a correlation between sub-clonal cluster expansion in the testis and elevated mutant sperm production in older donors, limited data exist on the universality of this phenomenon. Our study addresses this gap by examining the testis-expansion patterns, as well as the increases in mutations in sperm for two FGFR3 variants-c.1138G>A (p.G380R) and c.1948A>G (p.K650E)-which are associated with ACH or thanatophoric dysplasia (TDII), respectively. Unlike the ACH mutation, which showed sub-clonal expansion events in an aged testis and a significant increase in mutant sperm with the donor's age, as also reported in other studies, the TDII mutation showed focal mutation pockets in the testis but exhibited reduced transmission into sperm and no significant age-related increase. The mechanism behind this divergence remains unclear, suggesting potential pleiotropic effects of aberrant RTK signalling in the male germline, possibly hindering differentiation requiring meiosis. This study provides further insights into the transmission risks of micro-mosaics associated with advanced paternal age in the male germline.

Objective: To compare psychiatric emergencies and self-harm at emergency departments (EDs) 1 year into the pandemic, to early pandemic and pre-pandemic, and to examine the changes in the characteristics of self-harm presentations., Method: This retrospective cohort study expanded on the Pandemic-Related Emergency Psychiatric Presentations (PREP-kids) study. Routine record data in March to April of 2019, 2020, and 2021 from 62 EDs in 25 countries were included. ED presentations made by children and adolescents for any mental health reasons were analyzed., Results: Altogether, 8,174 psychiatric presentations were recorded (63.5% female; mean [SD] age, 14.3 [2.6] years), 3,742 of which were self-harm presentations. Rate of psychiatric ED presentations in March to April 2021 was twice as high as in March to April 2020 (incidence rate ratio [IRR], 1.93; 95% CI, 1.60-2.33), and 50% higher than in March to April 2019 (IRR, 1.51; 95% CI, 1.25-1.81). Rate of self-harm presentations doubled between March to April 2020 and March to April 2021 (IRR, 1.98; 95% CI, 1.68-2.34), and was overall 1.7 times higher than in March to April 2019 (IRR, 1.70; 95% CI, 1.44-2.00). Comparing self-harm characteristics in March to April 2021 with March to April 2019, self-harm contributed to a higher proportion of all psychiatric presentations (odds ratio [OR], 1.30; 95% CI, 1.05-1.62), whereas female representation in self-harm presentations doubled (OR, 1.98; 95% CI, 1.45-2.72) and follow-up appointments were offered 4 times as often (OR, 4.46; 95% CI, 2.32-8.58)., Conclusion: Increased pediatric ED visits for both self-harm and psychiatric reasons were observed, suggesting potential deterioration in child mental health. Self-harm in girls possibly increased and needs to be prioritized. Clinical services should continue using follow-up appointments to support discharge from EDs., Diversity & Inclusion Statement: One or more of the authors of this paper self-identifies as a member of one or more historically underrepresented racial and/or ethnic groups in science. We actively worked to promote inclusion of historically underrepresented racial and/or ethnic groups in science in our author group. While citing references scientifically relevant for this work, we also actively worked to promote inclusion of historically underrepresented racial and/or ethnic groups in science in our reference list. The author list of this paper includes contributors from the location and/or community where the research was conducted who participated in the data collection, design, analysis, and/or interpretation of the work., (Copyright © 2023 American Academy of Child and Adolescent Psychiatry. Published by Elsevier Inc. All rights reserved.)

Background: Children and young people's (CYP) mental health is worsening, and an increasing number are seeking psychiatric and mental health care. Whilst many CYPs with low-to-medium levels of psychiatric distress can be treated in outpatient services, CYPs in crisis often require inpatient hospital treatment. Although necessary in many cases, inpatient care can be distressing for CYPs and their families. Amongst other things, inpatient stays often isolate CYPs from their support networks and disrupt their education. In response to such limitations, and in order to effectively support CYPs with complex mental health needs, intensive community-based treatment models, which are known in this paper as intensive community care services (ICCS), have been developed. Although ICCS have been developed in a number of settings, there is, at present, little to no consensus of what ICCS entails., Methods: A group of child and adolescent mental health clinicians, researchers and academics convened in London in January 2023. They met to discuss and agree upon the minimum requirements of ICCS. The discussion was semi-structured and used the Dartmouth Assertive Community Treatment Fidelity Scale as a framework. Following the meeting, the agreed features of ICCS, as described in this paper, were written up., Results: ICCS was defined as a service which provides treatment primarily outside of hospital in community settings such as the school or home. Alongside this, ICCS should provide at least some out-of-hours support, and a minimum of 90% of CYPs should be supported at least twice per week. The maximum caseload should be approximately 5 clients per full time equivalent (FTE), and the minimum number of staff for an ICCS team should be 4 FTE. The group also confirmed the importance of supporting CYPs engagement with their communities and the need to remain flexible in treatment provision. Finally, the importance of robust evaluation utilising tools including the Children's Global Assessment Scale were agreed., Conclusions: This paper presents the agreed minimum requirements of intensive community-based psychiatric care. Using the parameters laid out herein, clinicians, academics, and related colleagues working in ICCS should seek to further develop the evidence base for this treatment model., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Fibroblast growth factor receptors (FGFRs) initiate signal transduction via the RAS/mitogen-activated protein kinase pathway by their tyrosine kinase activation known to determine cell growth, tissue differentiation, and apoptosis. Recently, many missense mutations have been reported for FGFR3, but we only know the functional effect for a handful of them. Some mutations result in aberrant FGFR3 signaling and are associated with various genetic disorders and oncogenic conditions. Here, we employed micropatterned surfaces to specifically enrich fluorophore-tagged FGFR3 (monomeric GFP [mGFP]-FGFR3) in certain areas of the plasma membrane of living cells. We quantified receptor activation via total internal reflection fluorescence microscopy of FGFR3 signaling at the cell membrane that captured the recruitment of the downstream signal transducer growth factor receptor-bound 2 (GRB2) tagged with mScarlet (GRB2-mScarlet) to FGFR3 micropatterns. With this system, we tested the activation of FGFR3 upon ligand addition (fgf1 and fgf2) for WT and four FGFR3 mutants associated with congenital disorders (G380R, Y373C, K650Q, and K650E). Our data showed that ligand addition increased GRB2 recruitment to WT FGFR3, with fgf1 having a stronger effect than fgf2. For all mutants, we found an increased basal receptor activity, and only for two of the four mutants (G380R and K650Q), activity was further increased upon ligand addition. Compared with previous reports, two mutant receptors (K650Q and K650E) had either an unexpectedly high or low activation state, respectively. This can be attributed to the different methodology, since micropatterning specifically captures signaling events at the plasma membrane. Collectively, our results provide further insight into the functional effects of mutations to FGFR3., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Non-suicidal-self-injury (NSSI) in adolescents needing inpatient treatment is a serious health risk behaviour. NSSI-specific treatment programs for inpatients hardly exist. "Cut the Cut" (CTC) is a new treatment program in intervals, addressing this problem. Aim of this pilot-study was to evaluate acceptability and feasibility of CTC. 23 female inpatients (12 CTC, 11 control, aged 15-17; mean = 16.80, SD.70) engaging in NSSI were evaluated for service user satisfaction, frequency, and severity of NSSI at T1 (admission), T2 (discharge after interval 1, CTC-group) and T3 (discharge). A qualitative interview was performed at T3. Significant improvement in NSSI-frequency was given (T1-T3: CTC p = 0.010; control p = 0.038). Severity of NSSI reduced slightly (mild NSSI: CTC p = 0.022, control p = 0.087; severe NSSI: CTC p = 0.111, control p = 0.066). Satisfaction of parents (T3 mean 28.38) and adolescents (T3 mean 26.11) in CTC was rated high. CTC is a feasible treatment option for inpatients engaging in NSSI. Further studies over time are needed.Trial registration Number DRKS00016762, 05.03.2019, retrospectively registered., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The mammary gland undergoes hormonally stimulated cycles of proliferation, lactation, and involution. We hypothesized that these factors increase the mutational burden in glandular tissue and may explain high cancer incidence rate in the general population, and recurrent disease. Hence, we investigated the DNA sequence variants in the normal mammary gland, tumor, and peripheral blood from 52 reportedly sporadic breast cancer patients. Targeted resequencing of 542 cancer-associated genes revealed subclonal somatic pathogenic variants of: PIK3CA, TP53, AKT1, MAP3K1, CDH1, RB1, NCOR1, MED12, CBFB, TBX3, and TSHR in the normal mammary gland at considerable allelic frequencies (9 × 10 -2 - 5.2 × 10 - 1 ), indicating clonal expansion. Further evaluation of the frequently damaged PIK3CA and TP53 genes by ultra-sensitive duplex sequencing demonstrated a diversified picture of multiple low-level subclonal (in 10 -2 -10 -4 alleles) hotspot pathogenic variants. Our results raise a question about the oncogenic potential in non-tumorous mammary gland tissue of breast-conserving surgery patients., (© 2022. The Author(s).)

De novo mutations (DNMs) are important players in heritable diseases and evolution. Of particular interest are highly recurrent DNMs associated with congenital disorders that have been described as selfish mutations expanding in the male germline, thus becoming more frequent with age. Here, we have adapted duplex sequencing (DS), an ultradeep sequencing method that renders sequence information on both DNA strands; thus, one mutation can be reliably called in millions of sequenced bases. With DS, we examined ∼4.5 kb of the FGFR3 coding region in sperm DNA from older and younger donors. We identified sites with variant allele frequencies (VAFs) of 10 -4 to 10 -5 , with an overall mutation frequency of the region of ∼6 × 10 -7 Some of the substitutions are recurrent and are found at a higher VAF in older donors than in younger ones or are found exclusively in older donors. Also, older donors harbor more mutations associated with congenital disorders. Other mutations are present in both age groups, suggesting that these might result from a different mechanism (e.g., postzygotic mosaicism). We also observe that independent of age, the frequency and deleteriousness of the mutational spectra are more similar to COSMIC than to gnomAD variants. Our approach is an important strategy to identify mutations that could be associated with a gain of function of the receptor tyrosine kinase activity, with unexplored consequences in a society with delayed fatherhood., (© 2022 Salazar et al.; Published by Cold Spring Harbor Laboratory Press.)

Background: Admission rates to child and adolescent mental health inpatient units in Germany are substantial (58.831 admissions in 2019). Historically, different treatment pathways have not been available. The evidence suggests that home treatment could be an alternative. The objective of this study was to assess the long-term stability of treatment gains among children and adolescents treated for serious mental health problems with home treatment (HT) as compared with inpatient treatment-as-usual (TAU)., Methods: Hundred patients were enrolled and randomized into an intervention (HT n = 54) and control group (TAU n = 46). Follow-up data were available after 8.4 months (n = 78) (T3) and after 4.3 years (n = 51) (T4). The primary outcome measured was overall level of functioning, for which the Children's Global Assessment Scale (CGAS) was used. Secondary outcomes included severity of patient impairment and parental competency in dealing with their child's symptoms, measured using the Health of the Nation Outcome Scales (HoNOSCA). A qualitative interview with parents was performed at T3 and T4., Results: Treatment effects remained stable for both groups at T3 and T4 (p < .001). After 4.3 years 70% of the parents in the intervention group stated satisfaction with the treatment received, while only 36.8% of the parents within the control group rated themselves as satisfied. 43.7% of parents in the home treatment group reported that the most helpful aspect of this pathway was the prompt and intensive professional help and advice they received as primary caregivers at home., Conclusions: Home treatment may be considered a viable alternative to inpatient care., (© 2021 The Authors. Child and Adolescent Mental Health published by John Wiley & Sons Ltd on behalf of Association for Child and Adolescent Mental Health.)

The SARS-CoV-2 pandemic has required the development of multiple testing systems to monitor and control the viral infection. Here, we developed a PCR test to screen COVID-19 infections that can process up to ~180 samples per day without the requirement of robotics. For this purpose, we implemented the use of multichannel pipettes and plate magnetics for the RNA extraction step and combined the reverse transcription with the qPCR within one step. We tested the performance of two RT-qPCR kits as well as different sampling buffers and showed that samples taken in NaCl or PBS are stable and compatible with different COVID-19 testing systems. Finally, we designed a new internal control based on the human RNase P gene that does not require a DNA digestion step. Our protocol is easy to handle and reaches the sensitivity and accuracy of the standardized diagnostic protocols used in the clinic to detect COVID-19 infections.

Mutations occurring during embryonic development affect only a subset of cells resulting in two or more distinct cell populations that are present at different levels, also known as postzygotic mosaicism (PZM). Although PZM is a common biological phenomenon, it is often overlooked as a source of disease due to the challenges associated with its detection and characterization, especially for very low-frequency variants. Moreover, PZM can cause a different phenotype compared to constitutional mutations. Especially, lethal mutations in receptor tyrosine kinase (RTK) pathway genes, which exist only in a mosaic state, can have completely new clinical manifestations and can look very different from the associated monogenic disorder. However, some key questions are still not addressed, such as the level of mosaicism resulting in a pathogenic phenotype and how the clinical outcome changes with the development and age. Addressing these questions is not trivial as we require methods with the sensitivity to capture some of these variants hidden away in very few cells. Recent ultra-accurate deep-sequencing approaches can now identify these low-level mosaics and will be central to understand systemic and local effects of mosaicism in the RTK pathway. The main focus of this review is to highlight the importance of low-level mosaics and the need to include their detection in studies of genomic variation associated with disease., (© 2020 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Provision of care for children with mental health disorders or behavioural difficulties differs from country to country. Historically, Germany has the highest number of inpatient beds throughout Europe (64 inpatient beds per 100,000 young people). In addition, nearly 146 departments for child and adolescent psychiatry offer day-care beds as well as ambulatory care. A high number of resident child and adolescent psychiatrists/psychotherapists (more than 12,000 professionals) complement provision of care in the different regions of Germany. Yet, only 50% of children and adolescents with mental health problems receive the treatment they need. Barriers to care - for example lack of transportation, fear of stigmatization, long distances in rural regions or fear of long hospital inpatient stays - keep families from presenting their child/adolescent to the relevant institutions. Recently, a new treatment module: StäB, an intensive daily home treatment, delivered by a multiprofessional team, has been added to the portfolio of treatment options in Germany. This closes the gap between highly intensive inpatient care and low-frequency outpatient treatment, allowing a continuum of care in intensity and frequency within the treatment alternatives., (© 2021 The Authors. Child and Adolescent Mental Health published by John Wiley & Sons Ltd on behalf of Association for Child and Adolescent Mental Health.)

The Development of Medication and Psychotherapy from 1952 to 1957 in the Child and Adolescent Psychiatric Clinics Weissenau and Gütersloh Abstract. Objective: As a result of recent discussions concerning the misuse of medication in children in postwar Germany, the paper analyzes the treatment methods, especially psychopharmacotherapy, as well as therapeutic approaches employed in two German child and adolescent psychiatric hospitals (Weissenau and Guetersloh). We investigated the progression of prescription and the administration of psychotropic and experimental drugs between 1952 and 1957. Methods: In total, 966 health records were extensively studied for the time period in question. Results: The use of psychotropic drugs was similar in both hospitals, yet even at that time the use of already established psychotropic drugs differed depending on the concept. Medication-induced fever and shock therapy via insulin were still in use in 1957. Occasionally, initial concepts of a therapeutic treatment approach were observed. Both institutions administered experimental drugs, albeit not in systematic clinical trials. Conclusion: The varying methods of treatment and medication strategies reflect the different setup and concept of the respective hospitals. Indications of dangerous medication trials were not detected. Keywords: child and adolescence psychiatry, psychotropic drugs, neuroleptics, experimental drugs, treatment methods.

Background: Interface management after inpatient care for mentally ill children and adolescents has been proven to be a breaking point in good transition of care between child and adolescent psychiatry, social welfare services, schools, job centre and the judicial system. Criteria for successful discharge management do not exist in child and adolescent psychiatry. Aim of the study ASpeKT was to survey parents on their perception of interface management and to derive recommendations for discharge management., Methods: Data regarding interface management were retrieved from parents (T3, n = 124, T4, n = 81) 6 months (T3) and 12 months (T4) after discharge., Results: The parents stated that accessible help after discharge from inpatient treatment is essential for stability and requires a good coordination. Parents named that they perceived helpful for successful interface management: a case manager, early round table meetings, support in returning to school, seamless access to outpatient follow-up appointments as well as information on further treatment options and contact data., Conclusion: From the perspective of affected families a proactive early individual and reliable care coordination by a constant contact person is essential for a good discharge management.

Duplex sequencing is currently the most reliable method to identify ultra-low frequency DNA variants by grouping sequence reads derived from the same DNA molecule into families with information on the forward and reverse strand. However, only a small proportion of reads are assembled into duplex consensus sequences (DCS), and reads with potentially valuable information are discarded at different steps of the bioinformatics pipeline, especially reads without a family. We developed a bioinformatics toolset that analyses the tag and family composition with the purpose to understand data loss and implement modifications to maximize the data output for the variant calling. Specifically, our tools show that tags contain polymerase chain reaction and sequencing errors that contribute to data loss and lower DCS yields. Our tools also identified chimeras, which likely reflect barcode collisions. Finally, we also developed a tool that re-examines variant calls from raw reads and provides different summary data that categorizes the confidence level of a variant call by a tier-based system. With this tool, we can include reads without a family and check the reliability of the call, that increases substantially the sequencing depth for variant calling, a particular important advantage for low-input samples or low-coverage regions., (© The Author(s) 2021. Published by Oxford University Press on behalf of NAR Genomics and Bioinformatics.)

Although aripiprazole is one of the most used antipsychotics, knowledge about serum concentrations in children and adolescents is scarce and age-specific therapeutic ranges have not been established yet. Data of a routine therapeutic drug monitoring service were analyzed in order to evaluate the relationship between dose and serum concentration of aripiprazole in children and adolescents. The study also aimed to evaluate whether the therapeutic reference range defined for adults with schizophrenia (100-350 ng/ml) is applicable for minors. Data from 130 patients (aged 7-19 years) treated with aripiprazole for different indications in doses of 2-30 mg/day were evaluated. Patient characteristics, doses, serum concentrations and therapeutic outcome were assessed by standardized measures. A positive mean correlation between body weight-corrected daily dose and aripiprazole concentration was found (r p  = 0.59, p < 0.001) with variation in dose explaining 35% of the variability in serum concentrations. Girls had on average 41% higher dose-corrected concentrations than boys (244.9 versus 173.4 mg/l; p = 0.006). Aripiprazole concentrations did not vary with co-medication (p = 0.22). About 70% of all measured serum concentrations were within the recommended therapeutic range for adults. Using a calculation method in all responding patients with an ICD-10 F2 diagnosis for a rough estimation of a preliminary therapeutic window also demonstrated a similar therapeutic range of aripiprazole in minors (105.9-375.3 ng/ml) than for adults. If confirmed in larger samples and more controlled study designs, these data may contribute to the definition of a therapeutic range of aripiprazole concentrations in children and adolescents.

From home treatment to psychiatric ward-equivalent treatment (StäB) - A systematic review of outpatient treatment in Germany Abstract. Objective: In the past, home treatment was not available in Germany for children and adolescents with mental health problems. However, enactment of the PsychVVG law on 1 January 2017 has changed this, so that intensive home treatment (stationsäquivalente Behandlung, StäB) can now be offered as an alternative to inpatient treatment. This literature review a) analyzes existing German home treatment studies for effectiveness, b) determines from international reviews criteria for effective home treatment, and c) aligns these with the standardized StäB criteria. Method: We performed a systematic literature search in PubMed, Scopus, PsycINFO, and Cochrane Library referencing "hometreatment AND children" as well as "hometreatment AND adolesc*. Results: Flexibility, mobility, multiprofessionality, a wide spectrum of possible interventions, the option of inpatient admission, 24/7 availability as well as daily treatment sessions have been found to be factors for effective home treatment. In German studies, home treatment was shown to enhance psychosocial functioning, parental competences, long-term effectiveness, and acceptance by the families. The highest effect for home treatment was demonstrated in children and adolescents with externalizing disorders. Conclusion: Intensive home treatment (StäB) meets the criteria identified in the literature and is a good addition to existing treatment options for children and adolescents with mental health problems in Germany.

Intensive outpatient treatment - a therapy option for all patients in child and adolescent psychiatry or just for a few? Abstract. Objective: "Child and adolescent psychiatric ward-equivalent treatment" (= stationsäquivalente Behandlung, StäB) is an intensive daily home treatment that has now become available in Germany. Conditions for StäB services were negotiated were negotiated, authorized and came into force by German healthcare commissioners and hospitals on 1 January 2018. However, to date few hospitals in Germany offer StäB, and its feasibility has been questioned. This study evaluates the first 58 cases of StäB in the Department of Child and Adolescent Psychiatry, ZfP Südwürttemberg (2018-2019). Method: All patients admitted consecutively to StäB from 1 January 2018 to 15 August 2019 were included and evaluated solely using descriptive analyses in SPSS.25. Results: The average length of stay in StäB was 37.95 days (SD 15.35). 86.2 % of the families completed treatment, with 3 patients discontinuing treatment and 5 patients needing inpatient admission. Unlike in the literature, internalising disorders predominated (70.7 %), whereas externalizing disorders were diagnosed in only 27.8 % of the patients. The cost of treatment in StäB was on average 8779.25 €. Conclusions: StäB is a viable and well-accepted new treatment alternative for patients requiring inpatient admission. The multiprofessionality of treatment as well as daily contacts are required. Feasibility under the conditions set is given. The average length of stay is slightly longer, but the costs are lower than those of inpatient treatment.

Background: Duplex sequencing is the most accurate approach for identification of sequence variants present at very low frequencies. Its power comes from pooling together multiple descendants of both strands of original DNA molecules, which allows distinguishing true nucleotide substitutions from PCR amplification and sequencing artifacts. This strategy comes at a cost-sequencing the same molecule multiple times increases dynamic range but significantly diminishes coverage, making whole genome duplex sequencing prohibitively expensive. Furthermore, every duplex experiment produces a substantial proportion of singleton reads that cannot be used in the analysis and are thrown away., Results: In this paper we demonstrate that a significant fraction of these reads contains PCR or sequencing errors within duplex tags. Correction of such errors allows "reuniting" these reads with their respective families increasing the output of the method and making it more cost effective., Conclusions: We combine an error correction strategy with a number of algorithmic improvements in a new version of the duplex analysis software, Du Novo 2.0. It is written in Python, C, AWK, and Bash. It is open source and readily available through Galaxy, Bioconda, and Github: https://github.com/galaxyproject/dunovo.

PRDM9 is a trans-acting factor directing meiotic recombination to specific DNA-binding sites by its zinc finger (ZnF) array. It was suggested that PRDM9 is a multimer; however, we do not know the stoichiometry or the components inducing PRDM9 multimerization. In this work, we used in vitro binding studies and characterized with electrophoretic mobility shift assays, mass spectrometry, and fluorescence correlation spectroscopy the stoichiometry of the PRDM9 multimer of two different murine PRDM9 alleles carrying different tags and domains produced with different expression systems. Based on the migration distance of the PRDM9-DNA complex, we show that PRDM9 forms a trimer. Moreover, this stoichiometry is adapted already by the free, soluble protein with little exchange between protein monomers. The variable ZnF array of PRDM9 is sufficient for multimerization, and at least five ZnFs form already a functional trimer. Finally, we also show that only one ZnF array within the PRDM9 oligomer binds to the DNA, whereas the remaining two ZnF arrays likely maintain the trimer by ZnF-ZnF interactions., (© 2019 Schwarz et al.)

As recombination plays an important role in evolution, its estimation and the identification of hotspot positions is of considerable interest. We propose a novel approach for estimating population recombination rates based on genotyping or sequence data that involves a sequential multiscale change point estimator. Our method also permits demography to be taken into account. It uses several summary statistics within a regression model fitted on suitable scenarios. Our proposed method is accurate, computationally fast, and provides a parsimonious solution by ensuring a type I error control against too many changes in the recombination rate. An application to human genome data suggests a good congruence between our estimated and experimentally identified hotspots. Our method is implemented in the R-package LDJump, which is freely available at https://github.com/PhHermann/LDJump., (© 2019 The Authors. Molecular Ecology Resources Published by John Wiley & Sons Ltd.)

Meiotic recombination has strong, but poorly understood effects on short tandem repeat (STR) instability. Here, we screened thousands of single recombinant products with sperm typing to characterize the role of polymorphic poly-A repeats at a human recombination hotspot in terms of hotspot activity and STR evolution. We show that the length asymmetry between heterozygous poly-A's strongly influences the recombination outcome: a heterology of 10 A's (9A/19A) reduces the number of crossovers and elevates the frequency of non-crossovers, complex recombination products, and long conversion tracts. Moreover, the length of the heterology also influences the STR transmission during meiotic repair with a strong and significant insertion bias for the short heterology (6A/7A) and a deletion bias for the long heterology (9A/19A). In spite of this opposing insertion-/deletion-biased gene conversion, we find that poly-A's are enriched at human recombination hotspots that could have important consequences in hotspot activation., (© 2019 Heissl et al.)

Background: Most psychiatric disorders in childhood and adolescence cause impairment in academic performance. Early interventions in school are thought to reduce the burden of disorder and prevent chronicity of disorder, while a delay in reachable help may result in more severe symptoms upon first time presentation, often then causing upon first-time presentation immediate need of inpatient care., Methods: The study aims at reducing hospitalization rates and increasing social participation and quality of life among children and adolescents by establishing collaborations between schools, mental health care services and youth welfare services. CCSchool offers children and adolescents, aged six to 18 years, who present with psychiatric problems associated to school problems, a standardized screening and diagnostic procedure as well as treatment in school if necessary. Students can participate in CCSchool in three federal states of Germany if they a) show symptoms vindicating a mental health diagnosis, b) present with confirmed school problems and c) have a level of general functioning below 70 on the children global assessment of Functioning (C-GAF). Intervention takes place in three steps: module A (expected n = 901, according to power calculation) with standardized diagnostic procedures; module B (expected n = 428) implies a school-based assessment followed by a first intervention; module C (expected n = 103) offering school-based interventions with either four to six sessions (basic, 80% of patients) or eight to 12 sessions (intensive, 20% of patients). Primary aim is to evaluate the effectiveness of CCSchool, in reducing the need of hospitalization in children with mental health problems. The analyses will be conducted by an independent institute using mainly data collected from patients and their caregivers during study participation. Additionally, claims data from statutory health insurances will be analysed. Relevant confounders will be controlled in all analyses., Discussion: Evaluation may show if CCSchool can prevent hospitalizations, enhance social participation and improve quality of life of children and adolescents with mental health problems by providing early accessible interventions in the school setting., Trial Registration: Deutsches Register Klinischer Studien, Trial registration number: DRKS00014838 , registered on 6th of June 2018.

Meiosis is initiated by a double-strand break (DSB) introduced in the DNA by a highly controlled process that is repaired by recombination. In many organisms, recombination occurs at specific and narrow regions of the genome, known as recombination hotspots, which overlap with regions enriched for DSBs. In recent years, it has been demonstrated that conversions and mutations resulting from the repair of DSBs lead to a rapid sequence evolution at recombination hotspots eroding target sites for DSBs. We still do not fully understand the effect of this erosion in the recombination activity, but evidence has shown that the binding of trans -acting factors like PRDM9 is affected. PRDM9 is a meiosis-specific, multi-domain protein that recognizes DNA target motifs by its zinc finger domain and directs DSBs to these target sites. Here we discuss the changes in affinity of PRDM9 to eroded recognition sequences, and explain how these changes in affinity of PRDM9 can affect recombination, leading sometimes to sterility in the context of hybrid crosses. We also present experimental data showing that DNA methylation reduces PRDM9 binding in vitro Finally, we discuss PRDM9-independent hotspots, posing the question how these hotspots evolve and change with sequence erosion.This article is part of the themed issue 'Evolutionary causes and consequences of recombination rate variation in sexual organisms'., (© 2017 The Authors.)

PR domain containing protein 9 (PRDM9) is a meiosis-specific, multi-domain protein that regulates the location of recombination hotspots by targeting its DNA recognition sequence for double-strand breaks (DSBs). PRDM9 specifically recognizes DNA via its tandem array of zinc fingers (ZnFs), epigenetically marks the local chromatin by its histone methyltransferase activity, and is an important tether that brings the DNA into contact with the recombination initiation machinery. A strong correlation between PRDM9-ZnF variants and specific DNA motifs at recombination hotspots has been reported; however, the binding specificity and kinetics of the ZnF domain are still obscure. Using two in vitro methods, gel mobility shift assays and switchSENSE, a quantitative biophysical approach that measures binding rates in real time, we determined that the PRDM9-ZnF domain forms a highly stable and long-lived complex with its recognition sequence, with a dissociation halftime of many hours. The ZnF domain exhibits an equilibrium dissociation constant (K D ) in the nanomolar (nM) range, with polymorphisms in the recognition sequence directly affecting the binding affinity. We also determined that alternative sequences (15-16 nucleotides in length) can be specifically bound by different subsets of the ZnF domain, explaining the binding plasticity of PRDM9 for different sequences. Finally, longer binding targets are preferred than predicted from the numbers of ZnFs contacting the DNA. Functionally, a long-lived complex translates into an enzymatically active PRDM9 at specific DNA-binding sites throughout meiotic prophase I that might be relevant in stabilizing the components of the recombination machinery to a specific DNA target until DSBs are initiated by Spo11.

In the intestine water has to be reabsorbed from the chymus across the intestinal epithelium. The osmolarity within the lumen is subjected to high variations meaning that water transport often has to take place against osmotic gradients. It has been hypothesized that LI-cadherin is important in this process by keeping the intercellular cleft narrow facilitating the buildup of an osmotic gradient allowing water reabsorption. LI-cadherin is exceptional among the cadherin superfamily with respect to its localization along the lateral plasma membrane of epithelial cells being excluded from adherens junction. Furthermore it has 7 but not 5 extracellular cadherin repeats (EC1-EC7) and a small cytosolic domain. In this study we identified the peptide VAALD as an inhibitor of LI-cadherin trans-interaction by modeling the structure of LI-cadherin and comparison with the known adhesive interfaces of E-cadherin. This inhibitory peptide was used to measure LI-cadherin dependency of water transport through a monolayer of epithelial CACO2 cells under various osmotic conditions. If LI-cadherin trans-interaction was inhibited by use of the peptide, water transport from the luminal to the basolateral side was impaired and even reversed in the case of hypertonic conditions whereas no effect could be observed at isotonic conditions. These data are in line with a recently published model predicting LI-cadherin to keep the width of the lateral intercellular cleft small. In this narrow cleft a high osmolarity can be achieved due to ion pumps yielding a standing osmotic gradient allowing water absorption from the gut even if the faeces is highly hypertonic.

To study meiotic recombination products, cis- or trans-association of disease polymorphisms, or allele-specific expression patterns, it is necessary to phase heterozygous polymorphisms separated by several kilobases. Haplotyping using long-range polymerase chain reaction (PCR) is a powerful, cost-effective method to directly obtain the phase of multiple heterozygous sites with standard laboratory equipment in a handful of loci for many samples. The method is based on the amplification of large genomic DNA regions (up to ~40 kb) with a reaction mixture that combines a proofreading polymerase with allele-specific primer pairs that preferentially amplify matched templates. The analysis of two heterozygous SNPs requires four reactions, each containing one of the four possible allele-specific primer combinations (two forward and two reverse primers), with the mismatches occurring at the 3' ends of the primers. The two correct primer combinations will more efficiently elongate the matching alleles than the alternative alleles, and the difference in amplification efficiency can be monitored with real-time PCR.

Characterizing polymorphisms on single molecules renders the phase of different alleles, and thus, haplotype information. Here, we describe a high-throughput method to genotype hundreds-of thousands single molecules in parallel using bead-emulsion haplotyping (BEH). Haplotyping via BEH is an emulsion-PCR-based method that was adapted to amplify multiple DNA fragments on paramagnetic, microscopic beads within a compartment formed by an aqueous-oil emulsion. This generates beads covered by thousands of clonal copies from several polymorphic regions of an initial DNA molecule that are then genotyped with fluorescently labeled probes. With BEH, up to three different polymorphisms (or more if several polymorphisms are within an amplicon) can be typed within a fragment of several kilobases in a singleexperiment, rendering haplotype information of a very large number of initial single molecules. The high throughput and digital nature of the method makes it ideal to quantify rare haplotypes or to assess the haplotype diversity in complex samples.

Real-time PCR-based genotyping methods, such as TaqMan allelic discrimination assays and allele-specific genotyping, are particularly useful when screening a handful of single nucleotide polymorphisms in hundreds of samples; either derived from different individuals, tissues, or pre-amplified DNA. Although real-time PCR-based methods such as TaqMan are well-established, alternative methods, like allele-specific genotyping, are powerful alternatives, especially for genotyping short tandem repeat (STR) length polymorphisms. Here, we describe all relevant aspects when developing an assay for a new SNP or STR using either TaqMan or allele-specific genotyping, respectively, such as primer and probe design, optimization of reaction conditions, the experimental procedure for typing hundreds of samples, and finally the data evaluation. Our goal is to provide a guideline for developing genotyping assays using these two approaches that render reliable and reproducible genotype calls involving minimal optimization.

The need in cancer research or evolutionary biology to detect rare mutations or variants present at very low frequencies (<10 -5 ) poses an increasing demand on lowering the detection limits of available methods. Here we demonstrated that amplifiable DNA lesions introduce important error sources in ultrasensitive technologies such as single molecule PCR (smPCR) applications (e.g. droplet-digital PCR), or next-generation sequencing (NGS) based methods. Using templates with known amplifiable lesions (8-oxoguanine, deaminated 5-methylcytosine, uracil, and DNA heteroduplexes), we assessed with smPCR and duplex sequencing that templates with these lesions were amplified very efficiently by proofreading polymerases (except uracil), leading to G->T, and to a lesser extent, to unreported G->C substitutions at 8-oxoguanine lesions, and C->T transitions in amplified uracil containing templates. Long heat incubations common in many DNA extraction protocols significantly increased the number of G->T substitutions. Moreover, in ∼50-80% smPCR reactions we observed the random amplification preference of only one of both DNA strands explaining the known 'PCR jackpot effect', with the result that a lesion became indistinguishable from a true mutation or variant. Finally, we showed that artifactual mutations derived from uracil and 8-oxoguanine could be significantly reduced by DNA repair enzymes., (© The Author 2016. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.)

In many countries hometreatment (HT) offers a cost-effective alternative to hospitalization for children and adolescents with mental health problems requiring intensive mental healthcare. However, the database on HT varies as HT may refer to different models and settings of intensive outpatient treatment. In Germany HT is not used routinely in mental healthcare in child and adolescent psychiatry, therefore the data on HT in Germany, especially in child and adolescent psychiatry, are scarce although funding for studies investigating the effectiveness of HT is available. This review represents a comprehensive search in electronic databases (1980-2014) of literature on HT. It provides as well an overview of the underlying concepts of and the present evidence for HT. In addition, the evidence base on HT for specific child and adolescent mental health disorders is reviewed. Future prospects for the development of HT in Germany facing the upcoming change in health service commissioning (PEPP = «pauschalierendes Entgeltsystem in Psychiatric und Psychosomatik>>) are discussed, as HT in child and adolescent psychiatry, when accurately indicated, can be a valid alternative to inpatient treatment.

Background: Nonsuicidal self-injury (NSSI) in adolescence has been described as comorbid condition in affective or anxiety disorders, as well as borderline personality disorder (BPD) and is a risk factor for later suicide attempts. Prevalence rates of NSSI decline steeply from adolescence to young adulthood. Yet, to the best of our knowledge, the longitudinal development of adolescent psychiatric patients with NSSI into their young adulthood has not been investigated. The aim of this study was to assess current NSSI and psychological impairment of young adults, who had been in treatment for NSSI in their adolescence., Methods: Former patients of the departments of child and adolescent psychiatry and psychotherapy in Ulm and Ravensburg, Germany (N = 52), who presented with NSSI in their adolescence, were recruited (average age: 21.5 years (SD = 2.6)). Data was assessed using questionnaires and structured clinical interviews. Two groups of participants with prevailing NSSI and ceased NSSI were compared concerning their current psychological impairment, history of NSSI, suicide attempts, and BPD diagnosis., Results: Around half of all participants had engaged in NSSI within the last year, and around half met diagnostic criteria for BPD. Although there was no significant association between current NSSI and BPD, an earlier age of onset of NSSI and a longer duration of NSSI during adolescence was significantly predictive of adult BPD. Two thirds of participants still met criteria of an axis 1 psychiatric disorder. Suicide attempts were reported by 53.8 % of all participants. Participants with current NSSI were more likely to meet criteria for a current axis 1 disorder, had engaged in NSSI more often in their lifetime, and reported more suicide attempts., Conclusions: Reduction of NSSI from adolescence to young adulthood was lower than described in previous community samples. This may be due to the initial high psychiatric impairment of this sample in adolescence. Early onset of NSSI seemed to be a risk factor for a longer duration of NSSI during adolescence but not for NSSI prevailing into adulthood. However, it was a risk factor for adult BPD. Furthermore, the occurrence of suicidal thoughts and behaviors and prevailing NSSI was highly associated.

Objective: Indications for home treatment for specific diagnoses in child and adolescent psychiatry have not yet been evaluated., Method: In a recent intervention study (primary outcome: length of stay), 92 patients aged 5 to 17 years were randomized into an intervention group (early discharge followed by home treatment in combination with inpatient treatment, where needed) and a control group (regular length inpatient treatment). The aim of this explorative analysis was to retrieve additional information on “what works for whom.” Outcome parameters were as follows: Children’s Global Assessment Scale (CGAS), Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA), Strengths and Difficulties Questionnaire (SDQ) and Columbia Impairment Scale (CIS) at T1 (within 14 days after intake), T2 (end of treatment) and T3 (∅ 8.4-month follow-up). Multiple regression was used to investigate the association between diagnoses, treatment setting, age, sex, and improvement in both groups., Results: In children externalizing disorders were predominant, whereas in adolescents internalizing disorders were prominent. Patients improved equally under both types of treatment. Home treatment, however, was rated by the patients to be significantly more effective in adolescents (SDQ p = .017), boys (CIS p = .009, SDQ p < .001), and with externalizing disorders (SDQ p = .005)., Conclusions: Home treatment may be considered an alternative to inpatient treatment, especially in boys with externalizing disorders.

Meiosis is a potentially important source of germline mutations, as sites of meiotic recombination experience recurrent double-strand breaks (DSBs). However, evidence for a local mutagenic effect of recombination from population sequence data has been equivocal, likely because mutation is only one of several forces shaping sequence variation. By sequencing large numbers of single crossover molecules obtained from human sperm for two recombination hotspots, we find direct evidence that recombination is mutagenic: Crossovers carry more de novo mutations than nonrecombinant DNA molecules analyzed for the same donors and hotspots. The observed mutations were primarily CG to TA transitions, with a higher frequency of transitions at CpG than non-CpGs sites. This enrichment of mutations at CpG sites at hotspots could predominate in methylated regions involving frequent single-stranded DNA processing as part of DSB repair. In addition, our data set provides evidence that GC alleles are preferentially transmitted during crossing over, opposing mutation, and shows that GC-biased gene conversion (gBGC) predominates over mutation in the sequence evolution of hotspots. These findings are consistent with the idea that gBGC could be an adaptation to counteract the mutational load of recombination.

Several studies have revealed that aquaporins play a role in tumor progression and invasion. In breast carcinomas, high levels of aquaporin 5 (AQP5), a membrane protein involved in water transport, have been linked to increased cell proliferation and migration, thus facilitating tumor progression. Despite the potential role of AQP5 in mammary oncogenesis, the mechanisms controlling mammary AQP5 expression are poorly understood. In other tissues, AQP5 expression has been correlated with its promoter methylation, yet, very little is known about AQP5 promoter methylation in the mammary gland. In this work, we used the mouse mammary gland cell line EpH4, in which we controlled AQP5 expression via the steroid hormone dexamethasone (Dex) to further investigate mechanisms regulating AQP5 expression. In this system, we observed a rapid drop of AQP5 mRNA levels with a delay of several hours in AQP5 protein, suggesting transcriptional control of AQP5 levels. Yet, AQP5 expression was independent of its promoter methylation, or to the presence of negative glucocorticoid receptor elements (nGREs) in its imminent promoter region, but was rather influenced by the cell proliferative state or cell density. We conclude that AQP5 promoter methylation is not a universal mechanism for AQP5 regulation and varies on cell and tissue type.

Background: Assessing youths in acute suicidal crisis is a common jet pivotal task in child and adolescent psychiatry, usually relying primarily on the clinicians skills of assessment. The objective of this pilot-study was to evaluate feasibility and usefulness of questionnaires during assessment of youths in acute suicidal crisis., Method: 31 adolescents, presenting for suicide assessment, and their caregivers, were asked upon emergency presentation to fill in the Suicidal-Ideation-Questionnaire (SIQ) and the Youth Life Status Questionnaire (Y-LSQ) before receiving an assessment by a clinician. The SIQ has 30 items, 8 of which are defined as critical items able to predict suicidality with the highest probability. The Y-LSQ (30 items) measures the overall level of psychological distress. It has one suicidal item, which was used in this study for validation of the SIQ result. Clinical judgment and test results were collected and analyzed by an independent researcher., Results: It was feasible to ask adolescents in acute suicidal crisis to fill in a questionnaire. Clinical assessment of suicidality did not correlate significantly with the overall SIQ-score (p = 0.089), however there was a significant correlation between the SIQ 8 critical item result and clinical judgement of suicidality (p = 0.050)., Conclusion: The 8 critical SIQ items can be used to support clinical judgment of suicidality in acute crisis.