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Duchenne muscular dystrophy (DMD) is typically caused by mutations that disrupt the

Background Ulcerative proctitis (UP) is an uncommon presentation in paediatric patients with ulcerative colitis, accounting for Methods A retrospective cohort study involving 37 sites affiliated with the IBD Interest group of ESPGHAN. Data were collected at different timepoints from patients diagnosed with UP aged Results Two hundred and fifty patients with UP were included, with a median age at diagnosis of 14.5 (IQR 12.3-15.9) years, and a median follow-up of 2.7 (IQR 1.7-3.9) years. The most common presenting symptoms were bloody stools (94%), abdominal pain (59%) and diarrhea (53%). At diagnosis, the median pediatric ulcerative colitis activity index (PUCAI) score was 25 (IQR 20-35), and only 3 patients (1.3%) presented with ASC. Most children had normal inflammatory markers and albumin levels. the median fecal calprotectin level was 720 mcg/g (IQR 310-1800), while 16 patients (9.9%) had a calprotectin level Conclusion Disease burden is significant in paediatric patients that present with UP, with high rates of proximal disease extension and requirement for treatment escalation.

Instead of combining data from an ensemble of different models after the simulations are already performed, as in a standard multi-model ensemble, we let the models interact with each other during their simulation. This ensemble of interacting models is called a supermodel. By exchanging information, models can compensate for each other's errors before the errors grow and spread to other regions or variables. Effectively, we create a new dynamical system. The exchange between the models is frequent enough such that the models synchronize, in order to prevent loss of variance when the models are combined. In previous work, we experimented successfully with combining atmospheric models of intermediate complexity in the context of parametric error. Here we will show results of combining two different AGCMs, NorESM1-ATM and CESM1-ATM. The models have different horizontal and vertical resolutions. To combine states from the different grids, we convert the individual model states to a ‘common state space’ with interpolation techniques. The weighted superposition of different model states is called a ‘pseudo-observation’. The pseudo-observations are assimilated back into the individual models, after which the models continue their run. We apply recently developed methods to train the weights determining the superposition of the model states, in order to obtain a supermodel that will outperform the individual models and any weighted average of their outputs.

The neural crest is a multipotent, highly migratory cell population that gives rise to diverse cell types, including melanocytes. Factors regulating the development of the neural crest and emigration of its cells are likely to influence melanoma metastasis. The transcription factor FOXD3 plays an essential role in premigratory neural crest development and has been implicated in melanoma cell dormancy and response to therapeutics. FOXD3 is downregulated during the migration of the melanocyte lineage from the neural crest, and our previous work supports a role for FOXD3 in suppressing melanoma cell migration and invasion. Alternatively, TWIST1 is known to have promigratory and proinvasive roles in a number of cancers, including melanoma. Using ChIP-seq analysis, TWIST1 was identified as a potential transcriptional target of FOXD3. Mechanistically, FOXD3 directly binds to regions of the TWIST1 gene locus, leading to transcriptional repression of TWIST1 in human mutant BRAF melanoma cells. In addition, depletion of endogenous FOXD3 promotes upregulation of TWIST1 transcripts and protein. Finally, FOXD3 expression leads to a significant decrease in cell migration that can be efficiently reversed by the overexpression of TWIST1. These findings uncover the novel interplay between FOXD3 and TWIST1, which is likely to be important in the melanoma metastatic cascade.Implications: FOXD3 and TWIST1 define distinct subgroups of cells within a heterogeneous tumor. Mol Cancer Res; 12(9); 1314–23. ©2014 AACR.

Supplementary Figures S1-S2. Supplementary Figure S1. 1205Lu cells were transfected with a control non-targeting siRNA or one of two different FOXD3 targeting siRNAs. Lysates and mRNA were harvested four days after siRNA transfection. In addition, control siRNA treated cells were treated for 24 hours with the RAF inhibitor PLX4720 (1 μM) as a positive control for FOXD3 expression. Western blots and quantitative RT-PCR were performed. Columns, average of three independent experiments; bars, SD; *p

Supplementary Figure 1 from Deletion of PTEN Promotes Tumorigenic Signaling, Resistance to Anoikis, and Altered Response to Chemotherapeutic Agents in Human Mammary Epithelial Cells

Supplementary Figure Legends 1-2 from Deletion of PTEN Promotes Tumorigenic Signaling, Resistance to Anoikis, and Altered Response to Chemotherapeutic Agents in Human Mammary Epithelial Cells

Supplementary Figure 2 from Deletion of PTEN Promotes Tumorigenic Signaling, Resistance to Anoikis, and Altered Response to Chemotherapeutic Agents in Human Mammary Epithelial Cells

PDF file - 3777K, TWIST1 expression is not regulated by RSK, STAT3, AKT, or by protein stability in mutant B-RAF cell lines

PDF file - 1510K, TWIST1 is positively regulated by active ERK1/2 signaling in N-RAS mutant melanoma cells

PDF file - 2984K, Oligonucleotide and primer sequences

PDF file - 1651K, Absence of EMT-like protein changes following altered TWIST1 expression in melanoma cells

PDF file - 3308K, High TWIST1 expression allows for melanoma spheroid outgrowth

Many cancers, including breast cancer, harbor loss-of-function mutations in the catalytic domain of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) or have reduced PTEN expression through loss of heterozygosity and/or epigenetic silencing mechanisms. However, specific phenotypic effects of PTEN inactivation in human cancer cells remain poorly defined without a direct causal connection between the loss of PTEN function and the development or progression of cancer. To evaluate the biological and clinical relevance of reduced or deleted PTEN expression, a novel in vitro model system was generated using human somatic cell knockout technologies. Targeted homologous recombination allowed for a single and double allelic deletion, which resulted in reduced and deleted PTEN expression, respectively. We determined that heterozygous loss of PTEN in the nontumorigenic human mammary epithelial cell line MCF-10A was sufficient for activation of the phosphoinositide 3-kinase/AKT and mitogen-activated protein kinase pathways, whereas the homozygous absence of PTEN expression led to a further increased activation of both pathways. The deletion of PTEN was able to confer growth factor–independent proliferation, which was confirmed by the resistance of the PTEN−/− MCF-10A cells to small-molecule inhibitors of the epidermal growth factor receptor. However, neither heterozygous nor homozygous loss of PTEN expression was sufficient to promote anchorage-independent growth, but the loss of PTEN did confer apoptotic resistance to cell rounding and matrix detachment. Finally, MCF-10A cells with the reduction or loss of PTEN showed increased susceptibility to the chemotherapeutic drug doxorubicin but not paclitaxel. [Cancer Res 2009;69(21):8275–83]

PDF file - 3105K, TWIST1 is required for melanoma cell migration towards and invasion through collagen type I

PDF file - 11564K, MMP-1 is able to rescue defects in spheroid outgrowth resulting from TWIST1 ablation

DMD pathogenic variants for Duchenne and Becker muscular dystrophy are detectable with high sensitivity by standard clinical exome analyses of genomic DNA. However, up to 7% of DMD mutations are deep intronic and analysis of muscle-derived RNA is an important diagnostic step for patients who have negative genomic testing but abnormal dystrophin expression in muscle. In this study, muscle biopsies were evaluated from 19 patients with clinical features of a dystrophinopathy, but negative clinical DMD mutation analysis. Reverse transcription-polymerase chain reactionor high-throughput RNA sequencingmethods identified 19 mutations with one of three pathogenic pseudoexon types: deep intronic point mutations, deletions or insertions, and translocations. In association with point mutations creating intronic splice acceptor sites, we observed the first examples of DMD pseudo 3'-terminal exon mutations causing high efficiency transcription termination within introns. This connection between splicing and premature transcription termination is reminiscent of U1 snRNP-mediating telescripting in sustaining RNA polymerase II elongation across large genes, such as DMD. We propose a novel classification of three distinct types of mutations identifiable by muscle RNA analysis, each of which differ in potential treatment approaches. Recognition and appropriate characterization may lead to therapies directed toward full-length dystrophin expression for some patients.

Fascioscapulohumeral muscular dystrophy (FSHD) is caused by a unique genetic mechanism that relies on contraction and hypomethylation of the D4Z4 macrosatellite array on the chromosome 4q telomere allowing ectopic expression of theDUX4gene in skeletal muscle. Genetic analysis is difficult due to the large size and repetitive nature of the array, a nearly identical array on the 10q telomere, and the presence of divergent D4Z4 arrays scattered throughout the genome. Here, we combine nanopore long-read sequencing with Cas9-targeted enrichment of 4q and 10q D4Z4 arrays for comprehensive genetic analysis including determination of the length of the 4q and 10q D4Z4 arrays with base-pair resolution. In the same assay, we differentiate 4q from 10q telomeric sequences, determine A/B haplotype, identify paralogous D4Z4 sequences elsewhere in the genome, and estimate methylation for all CpGs in the array. Asymmetric, length-dependent methylation gradients were observed in the 4q and 10q D4Z4 arrays that reach a hypermethylation point at approximately 10 D4Z4 repeat units, consistent with the known threshold of pathogenic D4Z4 contractions. High resolution analysis of individual D4Z4 repeat methylation revealed areas of low methylation near the CTCF/insulator region and areas of high methylation immediately preceding theDUX4transcriptional start site. Within theDUX4exons, we observed a waxing/waning methylation pattern with a 180-nucleotide periodicity, consistent with phased nucleosomes. Targeted nanopore sequencing complements recently developed molecular combing and optical mapping approaches to genetic analysis for FSHD by adding precision of the length measurement, base-pair resolution sequencing, and quantitative methylation analysis.

This research addresses the assessment of adipose tissue (AT) and spatial distribution of visceral (VAT) and subcutaneous fat (SAT) in the trunk from standardized magnetic resonance imaging at 3 T, thereby demonstrating the feasibility of deep learning (DL)–based image segmentation in a large population-based cohort in Germany (five sites). Volume and distribution of AT play an essential role in the pathogenesis of insulin resistance, a risk factor of developing metabolic/cardiovascular diseases. Cross-validated training of the DL-segmentation model led to a mean Dice similarity coefficient of >0.94, corresponding to a mean absolute volume deviation of about 22 ml. SAT is significantly increased in women compared to men, whereas VAT is increased in males. Spatial distribution shows age- and body mass index–related displacements. DL-based image segmentation provides robust and fast quantification of AT (≈15 s per dataset versus 3 to 4 hours for manual processing) and assessment of its spatial distribution from magnetic resonance images in large cohort studies.

Background and ObjectivesTo describe the phenotypic spectrum of dystrophinopathy in a large cohort of individuals with DMD exon 2 duplications (Dup2), who may be particularly amenable to therapies directed at restoring expression of either full-length dystrophin or nearly full-length dystrophin through utilization of the DMD exon 5 internal ribosome entry site (IRES).MethodsIn this retrospective observational study, we analyzed data from large genotype–phenotype databases (the United Dystrophinopathy Project [UDP] and the Italian DMD network) and classified participants into Duchenne muscular dystrophy (DMD), intermediate muscular dystrophy (IMD), or Becker muscular dystrophy (BMD) phenotypes. Log-rank tests for time-to-event variables were used to compare age at loss of ambulation (LOA) in participants with Dup2 vs controls without Dup2 in the UDP database and for comparisons between steroid-treated vs steroid-naive participants with Dup2.ResultsAmong 66 participants with Dup2 (UDP = 40, Italy = 26), 61% were classified as DMD, 9% as IMD, and 30% as BMD. Median age at last observation was 15.4 years (interquartile range 8.79–26.0) and 75% had been on corticosteroids for at least 6 months. Age at LOA differed significantly between participants with Dup2 DMD and historical controls without Dup2 DMD (p < 0.001). Valid spirometry was limited but suggested a delay in the typical age-related decline in forced vital capacity and 24 of 55 participants with adequate cardiac data had cardiomyopathy.DiscussionSome patients with Dup2 display a milder disease course than controls without Dup2 DMD, and prolonged ambulation with corticosteroids suggests the potential of IRES activation as a molecular mechanism. As Dup2-targeted therapies reach clinical applications, this information is critical to aid in the interpretation of the efficacy of new treatments.

BACKGROUND: Necrotizing enterocolitis (NEC) remains a major complication in the neonatal population. Standard practices regarding the care of premature infants and attitudes toward NEC prevention strategies vary across neonatal intensive care units (NICUs). Evidence-based best practice dissemination was presented through the NEC Zero bundle. To close gaps between evidence and practice, a telehealth-delivered intervention (ie, NeoECHO) was provided to NICUs. PURPOSE: The purpose of the study was to enable adoption of best practice for NEC prevention using NeoECHO through describing the local volunteer unit leaders’, or internal facilitators (IF), experiences in participating in NeoECHO and identifying the extent to which the facilitation activities within the NeoECHO experience were consistent with constructs from the integrated Promoting Action on Research Implementation in Health Services (iPARHIS) framework. METHODS: The design of the study was qualitative descriptive. Six internal facilitators (IFs) were recruited in the Southwest. After 6 NeoECHO sessions, individual interviews were conducted with each IF and transcribed verbatim. Content analysis was applied. Codes were informed by the iPARIHS framework. RESULTS: Major themes were (1) Innovation themes of quality improvement projects, Bundles of Care, and Huddle; (2) Recipient themes of Reluctant Stakeholders and Technical Modalities; (3) Context themes of Buy-In, Timing, Resources, Leadership, and Blame; (4) Facilitation themes of Betterment, Buddy System, Passionate Care, and Empowerment; and (5) Adoption themes of Continuous Quality Improvement (CQI), Evidence-Based Practice, and Honest Discussions. IMPLICATIONS FOR PRACTICE AND RESEARCH: NeoECHO fostered a learning community to share current practices, policies, and strategies to implement prevention, but the internal facilitators were essential to foster local participation. The long-term impacts of NeoECHO are the focus of current research.

This theoretical article revives a classical bridging construct, canalization, to describe a new model of a general factor of psychopathology. To achieve this, we have distinguished between two types of plasticity, an early one that we call 'TEMP' for 'Temperature or Entropy Mediated Plasticity', and another, we call 'canalization', which is close to Hebbian plasticity. These two forms of plasticity can be most easily distinguished by their relationship to 'precision' or inverse variance; TEMP relates to increased model variance or decreased precision, whereas the opposite is true for canalization. TEMP also subsumes increased learning rate, (Ising) temperature and entropy. Dictionary definitions of 'plasticity' describe it as the property of being easily shaped or molded; TEMP is the better match for this. Importantly, we propose that 'pathological' phenotypes develop via mechanisms of canalization or increased model precision, as a defensive response to adversity and associated distress or dysphoria. Our model states that canalization entrenches in psychopathology, narrowing the phenotypic state-space as the agent develops expertise in their pathology. We suggest that TEMP - combined with gently guiding psychological support - can counter canalization. We address questions of whether and when canalization is adaptive versus maladaptive, furnish our model with references to basic and human neuroscience, and offer concrete experiments and measures to test its main hypotheses and implications.

The U.S. Fish and Wildlife Service oversees the recovery of many species protected by the U.S. Endangered Species Act (ESA). Recent research suggests that a structured approach to allocating conservation resources could increase recovery outcomes for ESA listed species. Quantitative approaches to decision support can efficiently allocate limited financial resources and maximize desired outcomes. Yet, developing quantitative decision support under real-world constraints is challenging. Approaches that pair research teams and end-users are generally the most effective. However, co-development requires overcoming “hurdles” that can arise because of differences in the mental models of the co-development team. These include perceptions that: (1) scarce funds should be spent on action, not decision support; (2) quantitative approaches are only useful for simple decisions; (3) quantitative tools are inflexible and prescriptive black boxes; (4) available data are not good enough to support decisions; and (5) prioritization means admitting defeat. Here, we describe how we addressed these misperceptions during the development of a prototype resource allocation decision support tool for understanding trade-offs in U.S. endangered species recovery. We describe how acknowledging these hurdles and identifying solutions enabled us to progress with development. We believe that our experience can assist other applications of developing quantitative decision support for resource allocation.

Obtaining wellness and enhancing resilience will be increasingly more important for General Surgeons. Although these concepts are not new, the increased complexity of health care delivery has elevated the importance of these essential attributes. Instilling these practices should be emphasized during surgery residency and be modeled by surgical educators and surgeon leaders. The enhanced emphasis of wellness and resiliency is a positive step forward; however, more must be accomplished to ensure the well-being of a particularly group of vulnerable physicians. This chapter discusses the history and scientific theory behind wellness and resiliency, as well as practical suggestions for consideration.

BACKGROUND Caregivers in the neonatal intensive care unit (NICU) often determine care practices in silos, although access to learning communities can improve quality. Project ECHO, a telehealth-delivered mentoring intervention, provides specialists' expertise but not in the NICU until now. Necrotizing enterocolitis (NEC) prevention and timely recognition is one area where specialist support and engaging with a learning community could improve outcomes. NEC-Zero is one care bundle that aims to improve care quality by providing tools to implement NEC prevention in family-engaged ways. AIMS To examine the feasibility and acceptability of NeoECHO to disseminate NEC-Zero education and describe the intentions of internal facilitators (IFs) and clinicians to initiate quality improvement changes. METHODS This was a convergent mixed-methods study. Our team delivered the first neonatal adaptation of Project ECHO called "NeoECHO" to leverage facilitation as an implementation strategy to disseminate NEC-Zero evidence and support practice change. RESULTS Six IFs and seven NICUs participated. All units and IFs that began the series finished it. Of the 261 session attendees, 206 (79%) study evaluations were completed. Of those who completed evaluations, 89 (100%) completed at least one session and 29 (33%) completed three or more. Satisfaction was high. Participants appreciated the engaged and accessible format to learn from experts using real case examples and didactic sessions. Individuals and IFs reported intentions to adopt evidence based on NeoECHO. LINKING EVIDENCE TO ACTION NeoECHO was an acceptable and feasible way to engage under-resourced NICUs and share NEC-Zero evidence and tools. More research is needed to examine the impact of NeoECHO on care processes and patient outcomes.

Institutional anomie theory (IAT) attributes the relatively high levels of violent crime in the United States to a social structure characterized by institutional imbalance towards economic dominance and a cultural ethos known as the American Dream which emphasizes individual material success. One of the greatest challenges in testing IAT lies in how to measure these central structural and cultural constructs. While extant research has largely coalesced around the operationalization of institutional imbalance, the measurement of culture has been markedly inconsistent. Such inconsistency may account for the more equivocal support for its cultural dynamics relative to its institutional dynamics. Using data on 47 countries, this study compares several approaches to operationalizing the cultural ethos known as the American Dream and explores whether these different operationalizations produce consistent findings. Study results provide mixed support for the cultural dynamics of IAT regardless of which approach is used and indicate that the approach taken to the measurement of culture can result in entirely different conclusions. We discuss the implications of these findings for IAT and future research on the relationship between culture and crime.

Background The fecal metabolome is comprised from derivatives of the gut microbiome, the host, the environment, and the diet. Thus, it is a central output and effector that is linked with Crohn disease (CD). We aimed to characterize metabolomics profiles that are associated with CD and link metabolites with gut microbial diversity and composition. Methods Fecal metabolomics was performed using liquid chromatography coupled Mass Spectrometer (LC/MS). Microbiome was defined by 16Sseq. Multivariate analyses in MaAsLin2 identified differentially abundant metabolites and these were used for pathways enrichment analyses using Metacyc (PMID: 31586394). Hierarchical All-against-All association (HAllA; PMID: 35758795) was used to identify significant relationships between fecal metabolites and microbial amplicon sequence variants (ASVs). Results The cohort included 294 samples from 85 subjects (mean age 36, 51% males). 250 samples were from CD patients (n=59) and 44 samples were from controls (n=26). 545 metabolites were included in our predefined library and were subsequently analyzed. Multivariate analyses identified 86 increased and 18 decreased metabolites in CD, after controlling for age and gender (FDR Conclusion CD patients have distinct fecal metabolic signature which is also correlated with disease activity. Furthermore, we identified significant interactions between specific fecal metabolites and gut microbial ASVs enriched with saliva bacteria that were previously linked with CD. More research is needed to elucidate the factors governing this newly discovered link.

Despite a proliferation of virtual partial hospital programs (PHP) during the COVID-19 pandemic, there is a dearth of research on such programs. In the current study, we compared treatment outcomes and patient satisfaction between an in-person and a virtual PHP. Further, we examined patients' qualitative feedback about the virtual PHP.Participants included 282 patients attending a virtual PHP during the COVID-19 pandemic and 470 patients attending an in-person PHP one year prior. Patients completed daily measures of symptom severity, and post-treatment measures of patient satisfaction and treatment outcomes. Patients in the virtual PHP provided feedback about virtual care. Quantitative data were analyzed using multilevel modeling, and qualitative data were analyzed using the principles of inductive analysis.Patients experienced a reduction in depression (Our results suggest that virtual PHPs should be explored as an ongoing model of care that may help to systematically reduce barriers to accessing mental health services.

Background Despite increasing number of therapies available for patients with Crohn’s disease (CD), response rates are generally not high. Therefore, there is a great need to identify novel targets in patients with active disease. Methods Serum samples were obtained from pediatric patients with CD at the time of diagnosis and following induction therapy with exclusive enteral nutrition (EEN) or anti-TNFa therapy, and from control subjects, defined as having normal colonoscopy without past or present history of inflammatory bowel diseases or other immune-mediated disorder. Associated clinical and laboratory data from these subjects was also collected. Serums were subjected to Olink technology (metabolism panel), based on proximity extension assay, enabling analysis of 92 proteins in each sample. Analysis was performed on a Normalized Protein eXpression (NPX) file comparing effect of different factors on the overall biomarker expression by supervised, multivariate, principal component analysis and verification by univariate ANOVA with Benjamini-Hochberg and post-hoc Tukey analysis. Results Eighty-eight serum samples were collected: 30 from control subjects and 58 from 32 patients with CD. The median age of patients in the control and CD groups was 13.9 years (IQR 11.1–16.6) and 14.6 years (IQR 12.2–16.9), respectively (P=0.32). Twenty-four patients were treated with anti-TNFa agents and 8 with EEN. The median pediatric Crohn’s Disease Activity Index decreased from 35 (22.5–42.5) to 5 (0–12.5, P Conclusion Serum levels of ADGRE2 and ADGRG2 are differentially regulated in patients with active CD, and should further be validated in additional Cohorts. The expression patterns and function of these receptors on different immune cells should be defined in future studies.