Psychosocial support interventions for children and adolescents experiencing gender dysphoria or incongruence: a systematic review.

BACKGROUND: National and international guidelines recommend that psychosocial support should be a key component of the care offered to children and adolescents experiencing gender dysphoria/incongruence. However, specific approaches or interventions are not recommended. AIM: To identify and summarise evidence on the outcomes of psychosocial support interventions for children and adolescents (age 0-18) experiencing gender dysphoria/incongruence. METHODS: Systematic review and narrative synthesis. Database searches (MEDLINE; EMBASE; CINAHL; PsycINFO; Web of Science) were performed in April 2022, with results assessed independently by two reviewers. Peer-reviewed articles reporting the results of studies measuring outcomes of psychosocial support interventions were included. Quality was assessed using the Mixed Methods Appraisal Tool. RESULTS: Ten studies were included. Half were conducted in the US, with others from Australia, Canada, New Zealand and the UK. Six were pre-post analyses or cohort studies, three were mixed methods, and one was a secondary analysis of intervention data from four trials. Most studies were of low quality. Most analyses of mental health and psychosocial outcomes showed either benefit or no change, with none indicating negative or adverse effects. CONCLUSIONS: The small number of low-quality studies limits conclusions about the effectiveness of psychosocial interventions for children/adolescents experiencing gender dysphoria/incongruence. Clarity on the intervention approach as well as the core outcomes would support the future aggregation of evidence. More robust methodology and reporting is required. PROSPERO REGISTRATION NUMBER: CRD42021289659.

Impact of social transition in relation to gender for children and adolescents: a systematic review.

BACKGROUND: Increasing numbers of children and adolescents experiencing gender dysphoria or incongruence are being referred to specialist gender services. Historically, social transitioning prior to assessment was rare but it is becoming more common. AIM: To identify and synthesise studies assessing the outcomes of social transition for children and adolescents (under 18) experiencing gender dysphoria/incongruence. METHODS: A systematic review and narrative sythesis. Database searches (Medline, Embase, CINAHL, PsycINFO, Web of Science) were perfomed in April 2022. Studies reporting any outcome of social transition (full or partial) for children and adolescents experiencing gender dysphoria/incongruence were included. An adapted version of the Newcastle-Ottawa Scale for cohort studies was used to appraise study quality. RESULTS: Eleven studies were included (children (n=8) and adolescents (n=3)) and most were of low quality. The majority were from the US, featured community samples and cross-sectional analyses. Different comparator groups were used, and outcomes related to mental health and gender identity reported. Overall studies consistently reported no difference in mental health outcomes for children who socially transitioned across all comparators. Studies found mixed evidence for adolescents who socially transitioned. CONCLUSIONS: It is difficult to assess the impact of social transition on children/adolescents due to the small volume and low quality of research in this area. Importantly, there are no prospective longitudinal studies with appropriate comparator groups assessing the impact of social transition on mental health or gender-related outcomes for children/adolescents. Professionals working in the area of gender identity and those seeking support should be aware of the absence of robust evidence of the benefits or harms of social transition for children and adolescents. PROSPERO REGISTRATION NUMBER: CRD42021289659.

FOXA2 Cooperates with Mutant KRAS to Drive Invasive Mucinous Adenocarcinoma of the Lung.

The endoderm-lineage transcription factor FOXA2 has been shown to inhibit lung tumorigenesis in in vitro and xenograft studies using lung cancer cell lines. However, FOXA2 expression in primary lung tumors does not correlate with an improved patient survival rate, and the functional role of FOXA2 in primary lung tumors remains elusive. To understand the role of FOXA2 in primary lung tumors in vivo, here, we conditionally induced the expression of FOXA2 along with either of the two major lung cancer oncogenes, EGFRL858R or KRASG12D, in the lung epithelium of transgenic mice. Notably, FOXA2 suppressed autochthonous lung tumor development driven by EGFRL858R, whereas FOXA2 promoted tumor growth driven by KRASG12D. Importantly, FOXA2 expression along with KRASG12D produced invasive mucinous adenocarcinoma (IMA) of the lung, a fatal mucus-producing lung cancer comprising approximately 5% of human lung cancer cases. In the mouse model in vivo and human lung cancer cells in vitro, FOXA2 activated a gene regulatory network involved in the key mucous transcription factor SPDEF and upregulated MUC5AC, whose expression is critical for inducing IMA. Coexpression of FOXA2 with mutant KRAS synergistically induced MUC5AC expression compared with that induced by FOXA2 alone. ChIP-seq combined with CRISPR interference indicated that FOXA2 bound directly to the enhancer region of MUC5AC and induced the H3K27ac enhancer mark. Furthermore, FOXA2 was found to be highly expressed in primary tumors of human IMA. Collectively, this study reveals that FOXA2 is not only a biomarker but also a driver for IMA in the presence of a KRAS mutation. SIGNIFICANCE: FOXA2 expression combined with mutant KRAS drives invasive mucinous adenocarcinoma of the lung by synergistically promoting a mucous transcriptional program, suggesting strategies for targeting this lung cancer type that lacks effective therapies.

CRISPRi links COVID-19 GWAS loci to LZTFL1 and RAVER1.

BACKGROUND: To identify host genetic variants (SNPs) associated with COVID-19 disease severity, a number of genome-wide association studies (GWAS) have been conducted. Since most of the identified variants are located at non-coding regions, such variants are presumed to affect the expression of neighbouring genes, thereby influencing COVID-19 disease severity. However, it remains largely unknown which genes are influenced by such COVID-19 GWAS loci. METHODS: CRISPRi (interference)-mediated gene expression analysis was performed to identify genes functionally regulated by COVID-19 GWAS loci by targeting regions near the loci (SNPs) in lung epithelial cell lines. The expression of CRISPRi-identified genes was investigated using COVID-19-contracted human and monkey lung single-nucleus/cell (sn/sc) RNA-seq datasets. FINDINGS: CRISPRi analysis indicated that a region near rs11385942 at chromosome 3p21.31 (locus of highest significance with COVID-19 disease severity at intron 5 of LZTFL1) significantly affected the expression of LZTFL1 (P<0.05), an airway cilia regulator. A region near rs74956615 at chromosome 19p13.2 (locus located at the 3' untranslated exonic region of RAVER1), which is associated with critical illness in COVID-19, affected the expression of RAVER1 (P<0.05), a coactivator of MDA5 (IFIH1), which induces antiviral response genes, including ICAM1. The sn/scRNA-seq datasets indicated that the MDA5/RAVER1-ICAM1 pathway was activated in lung epithelial cells of COVID-19-resistant monkeys but not those of COVID-19-succumbed humans. INTERPRETATION: Patients with risk alleles of rs11385942 and rs74956615 may be susceptible to critical illness in COVID-19 in part through weakened airway viral clearance via LZTFL1-mediated ciliogenesis and diminished antiviral immune response via the MDA5/RAVER1 pathway, respectively. FUNDING: NIH.

CRISPRi-mediated functional analysis of NKX2-1-binding sites in the lung.

The transcription factor NKX2-1/TTF-1 is involved in lung pathophysiology, including breathing, innate defense and tumorigenesis. To understand the mechanism by which NKX2-1 regulates genes involved in such pathophysiology, we have previously performed ChIP-seq and identified genome-wide NKX2-1-binding sites, which revealed that NKX2-1 binds to not only proximal promoter regions but also multiple intra- and inter-genic regions of the genes regulated by NKX2-1. However, the roles of such regions, especially non-proximal ones, bound by NKX2-1 have not yet been determined. Here, using CRISPRi (CRISPR/dCas9-KRAB), we scrutinize the functional roles of 19 regions/sites bound by NKX2-1, which are located in genes involved in breathing and innate defense (SFTPB, LAMP3, SFTPA1, SFTPA2) and lung tumorigenesis (MYBPH, LMO3, CD274/PD-L1). Notably, the CRISPRi approach reveals that a portion of NKX2-1-binding sites are functionally indispensable while the rest are dispensable for the expression of the genes, indicating that functional roles of NKX2-1-binding sites are unequally yoked.

Place of death of children and young adults with a life-limiting condition in England: a retrospective cohort study.

OBJECTIVE: To assess trends in place of death for children with a life-limiting condition and the factors associated with death at home or hospice rather than hospital. DESIGN: Observational cohort study using linked routinely collected data. SETTING: England. PATIENTS: Children aged 0-25 years who died between 2003 and 2017. MAIN OUTCOME MEASURES: Place of death: hospital, hospice, home. Multivariable multinomial logistic regression models. RESULTS: 39 349 children died: 73% occurred in hospital, 6% in hospice and 16% at home. In the multivariable models compared with dying in a hospital: neonates were less likely, and those aged 1-10 years more likely, than those aged 28 days to <1 year to die in hospice. Children from all ethnic minority groups were significantly less likely to die in hospice, as were those in the most deprived group (RR 0.8, 95% CI 0.7 to 0.9). Those who died from 2008 were more likely than those who died earlier to die in a hospice.Children with cancer (RR 4.4, 95% CI 3.8 to 5.1), neurological (RR 2.0, 95% CI 1.7 to 2.3) or metabolic (RR 3.7, 95% CI 3.0 to 4.6) diagnoses were more likely than those with a congenital diagnosis to die in a hospice.Similar patterns were seen for clinical/demographic factors associated with home versus hospital deaths. CONCLUSIONS: Most children with a life-limiting condition continue to die in the hospital setting. Further research on preferences for place of death is needed especially in children with conditions other than cancer. Paediatric palliative care services should be funded adequately to enable equal access across all settings, diagnostic groups and geographical regions.

CRISPRi-mediated functional analysis of lung disease-associated loci at non-coding regions.

Genome-wide association studies have identified lung disease-associated loci; however, the functions of such loci are not well understood in part because the majority of such loci are located at non-coding regions. Hi-C, ChIP-seq and eQTL data predict potential roles (e.g. enhancer) of such loci; however, they do not elucidate the molecular function. To determine whether these loci function as gene-regulatory regions, CRISPR interference (CRISPRi; CRISPR/dCas9-KRAB) has been recently used. Here, we applied CRISPRi along with Hi-C, ChIP-seq and eQTL to determine the functional roles of loci established as highly associated with asthma, cystic fibrosis (CF), chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF). Notably, Hi-C, ChIP-seq and eQTL predicted that non-coding regions located at chromosome 19q13 or chromosome 17q21 harboring single-nucleotide polymorphisms (SNPs) linked to asthma/CF/COPD and chromosome 11p15 harboring an SNP linked to IPF interact with nearby genes and function as enhancers; however, CRISPRi indicated that the regions with rs1800469, rs2241712, rs12603332 and rs35705950, but not others, regulate the expression of nearby genes (single or multiple genes). These data indicate that CRISPRi is useful to precisely determine the roles of non-coding regions harboring lung disease-associated loci as to whether they function as gene-regulatory regions at a genomic level.

Hfq CLASH uncovers sRNA-target interaction networks linked to nutrient availability adaptation.

By shaping gene expression profiles, small RNAs (sRNAs) enable bacteria to efficiently adapt to changes in their environment. To better understand how Escherichia coli acclimatizes to nutrient availability, we performed UV cross-linking, ligation and sequencing of hybrids (CLASH) to uncover Hfq-associated RNA-RNA interactions at specific growth stages. We demonstrate that Hfq CLASH robustly captures bona fide RNA-RNA interactions. We identified hundreds of novel sRNA base-pairing interactions, including many sRNA-sRNA interactions and involving 3'UTR-derived sRNAs. We rediscovered known and identified novel sRNA seed sequences. The sRNA-mRNA interactions identified by CLASH have strong base-pairing potential and are highly enriched for complementary sequence motifs, even those supported by only a few reads. Yet, steady state levels of most mRNA targets were not significantly affected upon over-expression of the sRNA regulator. Our results reinforce the idea that the reproducibility of the interaction, not base-pairing potential, is a stronger predictor for a regulatory outcome.

Spontaneous psoas haematoma secondary to antiplatelet therapy: a rare cause of atraumatic hip pain.

A 92-year-old woman with previous total hip replacement presented with sudden onset of atraumatic hip pain and inability to weight bear. In the absence of radiographic signs of fracture, loosening or biochemical evidence of infection a CT scan of the pelvis and hips was performed, which showed psoas thickening. MRI identified two separate collections related to the psoas and greater trochanteric regions. Ultrasound-guided aspiration was performed to rule out infection and demonstrated a haematoma. In contrast to previously reported cases caused by anticoagulant therapy or inherited coagulopathy, this case was secondary to single antiplatelet agent treatment alone. In the increasingly co-morbid ageing population with associated polypharmacy, aspirin is a common primary and secondary prevention treatment. In patients with atraumatic hip pain, spontaneous psoas haematoma due to antiplatelet therapy should be considered and investigated appropriately.

Go ta catch 'em al or not enough time: Users motivations for playing Pokémon Go™ and non-users' reasons for not installing.

Urban exergames are played in the real-world environment using built-in mobile phone sensors. The influence of Pokémon Go on physical activity and sitting time has been examined previously, however limited research has explored motivations for using the application. Thus, the aim of this study was to explore Pokémon Go users' motivations for using the application, exusers' reasons for abandoning the game and non-users' reasons for not installing. After institutional ethical approval, the 'Physical Activity and Pokémon Go' questionnaire was developed using QualtricsTM and distributed using social media soon after launch in the United Kingdom (baseline). At baseline a total of 461 participants (n=193 male, n=265 female, n=3 transgender) who were predominantly white (n=420) and did not self-report a disability (n=443) completed the questionnaire. Users' (n=236) were questioned on their motivations for using Pokémon Go and non-users' provided reasons for not installing. At 3 months a total of 127 participants (n=23 users) completed the questionnaire again and all qualitative data was thematically analyzed. The most commonly reported reason for using Pokémon Go was 'to have fun' which was 86% and 83% at baseline and 3 months respectively. The second most frequent reason at baseline was 'friends were using it' (58%) and at 3 months was 'to be outside' (48%). The least common motivation for using Pokémon Go at both baseline and 3 months was 'to meet new people' (8% and 0% respectively). At baseline, social motives and competition were two general themes which encapsulated Pokémon Go users' motivation for using the application. There were two general themes reported by Pokémon Go users' as to why they did not think they would use the application in the future. These were application related factors and factors unrelated to Pokémon Go. Non-users reported a range of reasons for not using Pokémon Go, including lack of interest and a lack of time. Safety concerns and risk of adverse events were reported by both users and non-users. This is the first study to thematically analyze motives for using Pokémon Go in which the findings are: 1) future smartphone applications aiming to increase physical activity must ensure that objectives evolve to maintain initial interest and motivation to engage with applications; 2) game developers must consider the required phone storage and capability which could be a barrier to downloading; and 3) concerns of using the application including the safety of users and those around them.

Adeno-associated virus 2 bound to its cellular receptor AAVR.

Adeno-associated virus (AAV) is a leading vector for virus-based gene therapy. The receptor for AAV (AAVR; also named KIAA0319L) was recently identified, and the precise characterization of AAV-AAVR recognition is in immediate demand. Taking advantage of a particle-filtering algorithm, we report here the cryo-electron microscopy structure of the AAV2-AAVR complex at 2.8 Å resolution. This structure reveals that of the five Ig-like polycystic kidney disease (PKD) domains in AAVR, PKD2 binds directly to the spike region of the AAV2 capsid adjacent to the icosahedral three-fold axis. Residues in strands B and E, and the BC loop of AAVR PKD2 interact directly with the AAV2 capsid. The interacting residues in the AAV2 capsid are mainly in AAV-featured variable regions. Mutagenesis of the amino acids at the AAV2-AAVR interface reduces binding activity and viral infectivity. Our findings provide insights into the biology of AAV entry with high-resolution details, providing opportunities for the development of new AAV vectors for gene therapy.

The hit-and-return system enables efficient time-resolved serial synchrotron crystallography.

We present a 'hit-and-return' (HARE) method for time-resolved serial synchrotron crystallography with time resolution from milliseconds to seconds or longer. Timing delays are set mechanically, using the regular pattern in fixed-target crystallography chips and a translation stage system. Optical pump-probe experiments to capture intermediate structures of fluoroacetate dehalogenase binding to its ligand demonstrated that data can be collected at short (30 ms), medium (752 ms) and long (2,052 ms) intervals.

An EGFR ligand promotes EGFR-mutant but not KRAS-mutant lung cancer in vivo.

EGFR ligands (e.g., EGF and TGFA) have been shown to be clinically associated with poor survival in lung cancer. Since TGFA itself initiates autochthonous tumors in liver, breast, and pancreas but not in the lung in transgenic mice in vivo, it would appear that an EGFR ligand may not initiate but rather promote lung cancer. However, it has not been proven in vivo whether lung cancer is promoted by an EGFR ligand. Using transgenic mouse models conditionally expressing EGFRL858R or KrasG12D with TGFA (an EGFR ligand) in lung epithelium, we determined that TGFA promoted the growth of EGFRL858R-lung tumors in airway regions but not that of KrasG12D-lung tumors. Analysis of TCGA datasets identified ΔNp63 and AGR2 as potential key tumor-promoting regulators, which were highly induced in the TGFA-induced EGFRL858R-lung tumors. The expression of AGR2 was positively correlated with the expression of TGFA in human EGFR-mutant lung adenocarcinomas. The expression of TGFA in human EGFR-mutant lung adenocarcinomas but not in the EGFR wild-type lung adenocarcinoma was associated with poor survival. These results suggest that targeting EGFR ligands may benefit patients who carry EGFR-mutant lung tumors but will not benefit patients with KRAS-mutant lung tumors.

Validation of Three Postoperative Risk Prediction Models for Intensive Care Unit Mortality after Cardiac Surgery.

BACKGROUND: Several cardiac surgery risk prediction models based on postoperative data have been developed. However, unlike preoperative cardiac surgery risk prediction models, postoperative models are rarely externally validated or utilized by clinicians. The objective of this study was to externally validate three postoperative risk prediction models for intensive care unit (ICU) mortality after cardiac surgery. METHODS: The logistic Cardiac Surgery Scores (logCASUS), Rapid Clinical Evaluation (RACE), and Sequential Organ Failure Assessment (SOFA) scores were calculated over the first 7 postoperative days for consecutive adult cardiac surgery patients between January 2013 and May 2015. Model discrimination was assessed using receiver operating characteristic curve analyses. Calibration was assessed using the Hosmer-Lemeshow (HL) test, calibration plots, and observed to expected ratios. Recalibration of the models was performed. RESULTS: A total of 2255 patients were included with an ICU mortality rate of 1.8%. Discrimination for all three models on each postoperative day was good with areas under the receiver operating characteristic curve of >0.8. Generally, RACE and logCASUS had better discrimination than SOFA. Calibration of the RACE score was better than logCASUS, but ratios of observed to expected mortality for both were generally <0.65. Locally recalibrated SOFA, logCASUS and RACE models all performed well. CONCLUSION: All three models demonstrated good discrimination for the first 7 days after cardiac surgery. After recalibration, logCASUS and RACE scores appear to be most useful for daily risk prediction after cardiac surgery. If appropriately calibrated, postoperative cardiac surgery risk prediction models have the potential to be useful tools after cardiac surgery.

The impact of neurological disorders on hospital admissions for children and young people: a routine health data study.

INTRODUCTION: Neurological conditions are a major and increasing cause of hospitalisation among children and young people, but little is known about the impact of neurological conditions on hospital services in England, nor the factors that influence length of stay and bed days per year. OBJECTIVES: To quantify the hospital usage in children and young people related to neurological conditions, trends over time and variation by ethnicity and deprivation status. METHODS: An ICD10 coding framework identified a cohort of individuals aged 0-19 years with neurological conditions from linked routinely collected healthcare data from England (The Hospital Episode Statistics Admitted Patient Care dataset), from 1 April 2003 to 31 March 2015. Linked outpatient and accident and emergency data were used to supplement missing demographic data. Length of stay and bed days per year per person were calculated. These were separately modelled using random intercept multivariable negative binomial regressions with gender, age, ethnic group, diagnostic group, region of residence and deprivation category as predictors. RESULTS: 524,442 individuals were identified over the study period, increasing from 49,928 in 2003/04 to 102,840 in 2014/15. Neurological conditions account for 8.8% of inpatient bed days in the 0-14 year old age group. Length of stay and bed days per year vary primarily by age group - e.g. Under 1 year olds had 1.85 times (95%CI 1.83-1.86%) longer stays and over double (2.36 times, 95%CI 2.34-2.37 times) the number of bed days per person per year compared to 5 to 9 year olds - and main diagnostic group, with smaller variations by ethnic group, deprivation and region. CONCLUSIONS: Neurological conditions in children and young people have a significant and increasing impact on the NHS in England. Falls in length of stay and bed days per person are more than offset by increasing numbers of children and young people with neurological diagnoses. Variations in length of stay and bed days per year by diagnostic group, ethnic group, age group, deprivation category and region should be taken into account in resource planning.

Millimeter-wave emissivity as a metric for the non-contact diagnosis of human skin conditions.

A half-space electromagnetic model of human skin over the band 30-300 GHz was constructed and used to model radiometric emissivity. The model showed that the radiometric emissivity rose from 0.4 to 0.8 over this band, with emission being localized to a layer approximately one millimeter deep in the skin. Simulations of skin with differing water contents associated with psoriasis, eczema, malignancy, and thermal burn wounds indicated radiometry could be used as a non-contact technique to detect and monitor these conditions. The skin emissivity of a sample of 30 healthy volunteers, measured using a 95 GHz radiometer, was found to range from 0.2 to 0.7, and the experimental measurement uncertainty was ±0.002. Men on average were found to have an emissivity 0.046 higher than those of women, a measurement consistent with men having thicker skin than women. The regions of outer wrist and dorsal forearm, where skin is thicker, had emissivities 0.06-0.08 higher than the inner wrist and volar forearms where skin is generally thinner. Recommendations are made to develop a more sophisticated model of the skin and to collect larger data sets to obtain a deeper understanding of the signatures of human skin in the millimeter wave band. Bioelectromagnetics. 38:559-569, 2017. © 2017 The Authors. Bioelectromagnetics published by Wiley Periodicals, Inc.

Animal Models for Tendon Repair Experiments: A Comparison of Pig, Sheep and Human Deep Flexor Tendons in Zone II.

BACKGROUND: This laboratory study compared pig, sheep and human deep flexor tendons in regards to their biomechanical comparability. METHODS: To investigate the relevant biomechanical properties for tendon repair experiments, the tendons resistance to cheese-wiring (suture drag/splitting) was assessed. Cheese-wiring of a suture through a tendon is an essential factor for repair gapping and failure in a tendon repair. RESULTS: Biomechanical testing showed that forces required to pulling a uniform suture loop through sheep or pig tendons in Zone II were higher than in human tendons. At time point zero of testing these differences did not reach statistical significance, but differences became more pronounced when forces were measured beyond initial cheese-wiring (2 mm, 5 mm and 10 mm). The stronger resistance to cheese-wiring was more pronounced in the pig tendons. Also regarding size and histology, sheep tendons were more comparable to human tendons than pig tendons. CONCLUSIONS: Differences in tendon bio-properties should be kept in mind when comparing and interpreting the results of laboratory tendon experiments.