METHIONINE RESTRICTION INHIBITS NON-SMALL CELL LUNG CANCER GROWTH BY TARGETING THE BETA-CATENIN PATHWAY

Background: Lung cancer is the leading cause of cancer related death for both men and women. Non-small cell lung cancer (NSCLC) accounts for 80% of lung cancer with a 15% five-year survival rate. Current treatment options have serious side effects creating the need for alternative treatments. Methionine restriction (MR) has shown anti-tumor effects on various cancer cells, but the mechanisms involved in NSLCLC is unclear. The purpose of this study is to determine the anti-tumor effects of MR on NSCLC cells through the beta-catenin pathway. Objective: The purpose of this study is to determine the anti-tumor effects of MR on NSCLC cells through the beta-catenin pathway. Methods: Human NSCLC cell lines, A549 and H520 were obtained from ATCC and treated in the presence of normal or MR media (95% methionine restriction). After 48 hours of incubation, cell viability was determined by the alamar blue assay and a clonogenic assay was performed separately. A549 and H520 were treated for 24, 48, and 72 hours and cultured for harvest. Cell cycle was analyzed by measuring the DNA content of each cells determined using flow cytometry and western blot was performed using the antibodies β-actin, β-catenin, phospho β-catenin, and PARP. In order to investigate the potential molecular mechanism of MR on NSCLC cell, a human phospho-kinase array was performed. Results: MR significantly inhibits the cell proliferation in A549 and H520 cells after 48 hours. MR induces cell cycle arrest in G1 compared with the control after 24 hours of treatment. Protein expressions of PARP and phospho β-catenin are reduced in response to MR. The protein kinase array indicates MR exerts its anti-cancer effects by reducing phosphorylation of beta-catenin. Conclusion: Our results show MR has an inhibitory effect on the cell proliferation and colony formation of A549 and H520 cancer cell lines. Cell cycle arrest and reduced phosphorylated β-catenin provides insight into how methionine metabolism inhibits lung cancer development and progression. Further in vivo studies are needed in order to testify the efficacy of MR as a cancer prevention approach for NSCLC

‘Grounding a PIE in the sky’:Laying empirical foundations for a psychologically informed environment (PIE) to enhance well-being and practice in a homeless organisation

While psychologically informed environments (PIEs) are gaining in prominence in efforts to improve well-being and practice in the homeless sector, their empirical foundations remain tenuous. We present a unique scoping needs analysis of staff and client well-being, staff attitudes and the social–therapeutic climate in a UK-based homeless prevention organisation (prior to PIE implementation). Our aims were: (a) to apply a robust framework to pinpoint need and target forthcoming PIE initiatives and (b) to establish a validated needs baseline that informs and measures efficacy of PIE for its future development. Four established personal and practice well-being measures were administered to 134 (predominantly ‘frontline’) staff and 50 clients. Staff completed the: Warwick-Edinburgh Mental Wellbeing Scale (WEMWBS), Professional Quality of Life Scale (measuring compassion satisfaction [CS], burnout [BO] and secondary traumatic stress [STS]), Attitudes related to Trauma-informed Care Scale (ARTIC-10; measuring practice attitudes towards trauma-informed values) and the Essen Climate Evaluation Schema (EssenCES; measuring perceptions of client cohesion, safety and practitioner relationships in housing projects). Clients completed the WEMWBS and EssenCES. Vulnerability to STS was evident in nearly two-thirds of frontline staff and it was a statistically significant predictor of BO. It was not, however, associated with lesser levels of CS. We discuss this complex dynamic in relation to highlighted strategic recommendations for the PIE framework, and the identified potential challenges in implementing trauma-informed and reflective practice in the organisation. We conclude with a critique of the value and the lessons learnt from our efforts to integrate stronger empirical substance into the PIE approach

Refining the Primrose syndrome phenotype: A study of five patients with ZBTB20 de novo variants and a review of the literature

Primrose syndrome is a rare autosomal dominant condition caused by heterozygous missense variants within ZBTB20. Through an exome sequencing approach (as part of the Deciphering Developmental Disorders [DDD] study) we have identified five unrelated individuals with previously unreported, de novo ZBTB20 pathogenic missense variants. All five missense variants targeted the C2H2 zinc finger domains. This genotype‐up approach has allowed further refinement of the Primrose syndrome phenotype. Major characteristics (>90% individuals) include an intellectual disability (most frequently in the moderate range), a recognizable facial appearance and brain MRI abnormalities, particularly abnormalities of the corpus callosum. Other frequent clinical associations (in 50–90% individuals) include sensorineural hearing loss (83%), hypotonia (78%), cryptorchidism in males (75%), macrocephaly (72%), behavioral issues (56%), and dysplastic/hypoplastic nails (57%). Based upon these clinical data we discuss our current management of patients with Primrose syndrom

Understanding de novo onset of anxiety during COVID-19: Pre-pandemic socio-emotional functioning in vulnerable children

Background There is a need to understand and mitigate the psychological impacts of the COVID-19 pandemic for children known to be vulnerable. Data from prior to the pandemic are required to provide robust assessments of the socio-emotional impacts of COVID-19 and identify those who are more vulnerable. Method This study capitalises on an ongoing UK study of primary school children (4–8 years) identified prior to the pandemic as “at risk” for mental health problems by teachers. We collected mental health and social-emotional functioning data prior to the pandemic (Time 1) and re-assessed this cohort (N = 143) via researcher-led videocalls during lockdown (Time 2, summer 2020) and post-lockdown, 12 months later (Time 3; summer 2021). Results Mental health problems, particularly clinically significant anxiety, increased from 34% to 43% during lockdown and to 48% post-lockdown. Parental mental health difficulties (anxiety and depression) were prevalent during lockdown (40%) but had decreased 1 year later (20%). Children who developed clinically significant anxiety during the pandemic had impaired socio-emotional functioning at Time 1 (i.e., impaired emotion recognition, low self-esteem and social problems) and a high proportion (44%) had no contact with any peers during lockdown, which may have contributed to their anxiety, especially their school anxiety. Conclusion The pandemic appears to have exacerbated anxiety in already vulnerable children. A profile of socio-emotional problems identified a group of children who developed significant anxieties during the pandemic. These socio-emotional processes can be targeted for intervention to mitigate the negative mental health consequences of the pandemic and contribute to resilience in children

Family cluster of three cases of monkeypox imported from Nigeria to the United Kingdom, May 2021

Monkeypox is a rare viral zoonotic disease. The causing virus belongs to the Orthopoxvirus genus that includes variola virus (the cause of smallpox), vaccinia virus (used in the smallpox vaccine), and cowpox virus. There are two distinct clades of monkeypox virus—Central African and West African. The first human cases were identified in the Democratic Republic of Congo in 1970 [1]. Outside of Africa, cases of human monkeypox infections have been documented in four countries: four cases the United Kingdom (UK) in 2018/2019, one case in Israel in 2018 and one case in Singapore in 2019 [2], 47 cases in the United States (US) in 2003 and one in 2021 [3]. We report on a family cluster of three recent cases of monkeypox in the UK associated with travel from Nigeria

Expanding the genotypic spectrum of TXNL4A variants in Burn‐McKeown syndrome

From Wiley via Jisc Publications RouterHistory: received 2021-09-06, rev-recd 2021-10-21, accepted 2021-10-23, pub-electronic 2021-11-05Article version: VoRPublication status: PublishedFunder: Biotechnology and Biological Sciences Research Council; Id: http://dx.doi.org/10.13039/501100000268; Grant(s): BB/N000358/1Funder: Health Education England Genomics Education ProgrammeFunder: Medical Research Council; Id: http://dx.doi.org/10.13039/501100007155; Grant(s): 1916606Funder: National Institute for Health Manchester Biomedical Research Centre; Grant(s): IS‐BRC‐1215‐20007Abstract: The developmental disorder Burn‐McKeown Syndrome (BMKS) is characterised by choanal atresia and specific craniofacial features. BMKS is caused by biallelic variants in the pre‐messenger RNA splicing factor TXNL4A. Most patients have a loss‐of‐function variant in trans with a 34‐base pair (bp) deletion (type 1 Δ34) in the promoter region. Here, we identified two patients with BMKS. One individual has a TXNL4A c.93_94delCC, p.His32Argfs *21 variant combined with a type 1 Δ34 promoter deletion. The other has an intronic TXNL4A splice site variant (c.258‐3C>G) and a type 1 Δ34 promoter deletion. We show the c.258‐3C>G variant and a previously reported c.258‐2A>G variant, cause skipping of the final exon of TXNL4A in a minigene splicing assay. Furthermore, we identify putative transcription factor binding sites within the 56 bp of the TXNL4A promoter affected by the type 1 and type 2 Δ34 and use dual luciferase assays to identify a 22 bp repeated motif essential for TXNL4A expression within this promoter region. We propose that additional variants affecting critical transcription factor binding nucleotides within the 22 bp repeated motif could be relevant to BMKS aetiology. Finally, our data emphasises the need to analyse the non‐coding sequence in individuals where a single likely pathogenic coding variant is identified in an autosomal recessive disorder consistent with the clinical presentation

Prevalence and architecture of de novo mutations in developmental disorders.

The genomes of individuals with severe, undiagnosed developmental disorders are enriched in damaging de novo mutations (DNMs) in developmentally important genes. Here we have sequenced the exomes of 4,293 families containing individuals with developmental disorders, and meta-analysed these data with data from another 3,287 individuals with similar disorders. We show that the most important factors influencing the diagnostic yield of DNMs are the sex of the affected individual, the relatedness of their parents, whether close relatives are affected and the parental ages. We identified 94 genes enriched in damaging DNMs, including 14 that previously lacked compelling evidence of involvement in developmental disorders. We have also characterized the phenotypic diversity among these disorders. We estimate that 42% of our cohort carry pathogenic DNMs in coding sequences; approximately half of these DNMs disrupt gene function and the remainder result in altered protein function. We estimate that developmental disorders caused by DNMs have an average prevalence of 1 in 213 to 1 in 448 births, depending on parental age. Given current global demographics, this equates to almost 400,000 children born per year

Finding Diagnostically Useful Patterns in Quantitative Phenotypic Data.

Trio-based whole-exome sequence (WES) data have established confident genetic diagnoses in ∼40% of previously undiagnosed individuals recruited to the Deciphering Developmental Disorders (DDD) study. Here we aim to use the breadth of phenotypic information recorded in DDD to augment diagnosis and disease variant discovery in probands. Median Euclidean distances (mEuD) were employed as a simple measure of similarity of quantitative phenotypic data within sets of ≥10 individuals with plausibly causative de novo mutations (DNM) in 28 different developmental disorder genes. 13/28 (46.4%) showed significant similarity for growth or developmental milestone metrics, 10/28 (35.7%) showed similarity in HPO term usage, and 12/28 (43%) showed no phenotypic similarity. Pairwise comparisons of individuals with high-impact inherited variants to the 32 individuals with causative DNM in ANKRD11 using only growth z-scores highlighted 5 likely causative inherited variants and two unrecognized DNM resulting in an 18% diagnostic uplift for this gene. Using an independent approach, naive Bayes classification of growth and developmental data produced reasonably discriminative models for the 24 DNM genes with sufficiently complete data. An unsupervised naive Bayes classification of 6,993 probands with WES data and sufficient phenotypic information defined 23 in silico syndromes (ISSs) and was used to test a "phenotype first" approach to the discovery of causative genotypes using WES variants strictly filtered on allele frequency, mutation consequence, and evidence of constraint in humans. This highlighted heterozygous de novo nonsynonymous variants in SPTBN2 as causative in three DDD probands

International genome-wide meta-analysis identifies new primary biliary cirrhosis risk loci and targetable pathogenic pathways.

Primary biliary cirrhosis (PBC) is a classical autoimmune liver disease for which effective immunomodulatory therapy is lacking. Here we perform meta-analyses of discovery data sets from genome-wide association studies of European subjects (n=2,764 cases and 10,475 controls) followed by validation genotyping in an independent cohort (n=3,716 cases and 4,261 controls). We discover and validate six previously unknown risk loci for PBC (Pcombined<5 × 10(-8)) and used pathway analysis to identify JAK-STAT/IL12/IL27 signalling and cytokine-cytokine pathways, for which relevant therapies exist