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Background and Purpose Mutations in KCNQ3 have classically been associated with benign familial neonatal and infantile seizures and more recently identified in patients with neurodevelopmental disorders and abnormal electroencephalogram (EEG) findings. We present 4 affected patients from a family with a pathogenic mutation in KCNQ3 with a unique constellation of clinical findings. Methods A family of 3 affected siblings and mother sharing a KCNQ3 pathogenic variant are described, including clinical history, genetic results, and EEG and magnetic resonance imaging (MRI) findings. Results This family shows a variety of clinical manifestations, including neonatal seizures, developmental delays, autism spectrum disorder, and anxiety. One child developed absence epilepsy, 2 children have infrequent convulsive seizures that have persisted into childhood, and their parent developed adult-onset epilepsy. An underlying c.1091G>A (R364H) variant in KCNQ3 was found in all affected individuals. Conclusions The phenotypic variability of KCNQ3 channelopathies continues to expand as more individuals and families are described, and the variant identified in this family adds to the understanding of the manifestations of KCNQ3-related disorders.

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PDF file, 256K, Thalidomide effect on overall and progression-free survival according to glucocorticoid receptor (NR3C1) expression level.

PDF file, 142K, Effect of cumulative thalidomide dosing prior to start of maintence on OS and PFS, landmarked at start of maintence, for all patients.

PDF file, 94K, Post-relapse survival according to NR3C1 levels at baseline and at relapse.

PDF file, 297K, Survival outcomes in Total Therapy 2 according to randomization to control arm (-T) or thalidomide (+T).

Purpose: Because dexamethasone remains a key component of myeloma therapy, we wished to examine the impact of baseline and relapse expression levels of the glucocorticoid receptor gene NR3C1 on survival outcomes in the context of treatment with or without thalidomide.Experimental Design: We investigated the clinical impact of gene expression profiling (GEP)–derived expression levels of NR3C1 in 351 patients with GEP data available at baseline and in 130 with data available at relapse, among 668 subjects accrued to total therapy 2 (TT2).Results: Low NR3C1 expression levels had a negative impact on progression-free survival (PFS; HR, 1.47; P = 0.030) and overall survival (OS; HR, 1.90; P = 0.002) in the no-thalidomide arm. Conversely, there was a significant clinical benefit of thalidomide for patients with low receptor levels (OS: HR, 0.54; P = 0.015; PFS: HR, 0.54; P = 0.004), mediated most likely by thalidomide's upregulation of NR3C1. In the context of both baseline and relapse parameters, post-relapse survival (PRS) was adversely affected by low NR3C1 levels at relapse in a multivariate analysis (HR, 2.61; P = 0.012).Conclusion: These findings justify the inclusion of NR3C1 expression data in the work-up of patients with myeloma as it can significantly influence the choice of therapy and, ultimately, OS. The identification of an interaction term between thalidomide and NR3C1 underscores the importance of pharmacogenomic studies in the systematic study of new drugs. Clin Cancer Res; 18(19); 5499–506. ©2012 AACR.

PDF file, 56K, Salvage Regimen for 255 Patients with available data.

PDF file, 136K, Kaplan-Meier plots for overall survival from start of first salvage therapy.

Government funding supports industry-academia research and innovation projects in Norway, sharing the risk of the research component in innovation. However, funding alone may not be sufficient to overcome potential differences in collaborative agendas and ways of working. As a result, positive research outcomes often get stuck in the valley of death, instead of ending up as successful innovations that create value. To contribute to bridging the valley of death, we investigated the importance of six agile principles for collaborative industry-academia research and innovation projects, abbreviated IPN in Norway. The study was limited to the manufacturing sector. We surveyed 124 IPN project leaders (70 from industry; 54 from academia) to evaluate the importance of the knowledge management practices associated with the six agile principles across the three project stages. The statistical analyses indicate the consistency of the agile principles throughout the project stages. This means that agile principles are relevant for IPN projects and can be used as guidelines for improvement of the knowledge management practices. Moreover, the study identifies the agile principles that are perceived as most important to use in different stages of a project. It also identifies the different perceptions of the importance of agile principles of the project leaders from industry and academia. These findings can support project leaders who are implementing agile principles to industry-academia research and innovation projects. The results from the study can also support national and federal research/innovation councils in decision-making when assessing industrial research applications.

How do women experience breastfeeding multiples? Given the rising rate of multiple births and the global public health target of increasing the number of women exclusively breastfeeding up to the first 6 months, it is imperative that we understand why women who give birth to multiple babies breastfeed for shorter durations compared to those who have one baby. This chapter will explore the qualitative experiences of mothers who breastfeed twins/triplets and the social and physical capital women use to support multiple breastfeeding. Paying close attention to the mothers’ personal accounts this chapter will detail the many resources women draw on to meet the challenges of breastfeeding twins and triplets.

Using a qualitative design this study aimed to (1) explore the experience of people living with osteoarthritis (OA), (2) gain an understanding of their navigation of the health system and, (3) explore their opinions on the role of exercise and joint replacement surgery for the management of OA.Purposive sampling was used to recruit 26 participants with knee OA, aged 45 years and over, from Tasmania, Australia. Semi-structured interviews were audio-recorded, transcribed, coded, and thematically analysed to document participant understanding and experience of OA and their opinions on the role of exercise and surgery in managing OA.Of the 26 participants, 80% (n = 21) were female with a mean age of 66 years. The main theme identified was that individuals with knee OA were navigating a maze of OA treatments. Three related subthemes were that participants: (i) perceived their general practitioner did not have an ongoing role in their OA care, (ii) self-directed their management and, (iii) sampled from a 'smorgasbord' of treatment options, including low-value care options. Two other major themes were: the role of exercise for OA management, and surgery as a last resort.Our findings suggest that OA patients may not be choosing consistent, high-value care for their OA. This highlights the importance of an evidence-based multi-disciplinary approach to guide patients to self-manage their OA and support their navigation of the health system. Reducing emphasis on the pathway to surgery and streamlining access to conservative management strategies may assist people to receive high-value care.

Background Increased inflammation has been well defined in coronavirus disease 2019 (COVID-19), while definitive pathways driving severe forms of this disease remain uncertain. Neutrophils are known to contribute to immunopathology in infections, inflammatory diseases, and acute respiratory distress syndrome, a primary cause of morbidity and mortality in COVID-19. Changes in neutrophil function in COVID-19 may give insight into disease pathogenesis and identify therapeutic targets. Methods Blood was obtained serially from critically ill COVID-19 patients for 11 days. Neutrophil extracellular trap formation (NETosis), oxidative burst, phagocytosis, and cytokine levels were assessed. Lung tissue was obtained immediately postmortem for immunostaining. PubMed searches for neutrophils, lung, and COVID-19 yielded 10 peer-reviewed research articles in English. Results Elevations in neutrophil-associated cytokines interleukin 8 (IL-8) and interleukin 6, and general inflammatory cytokines IFN-inducible protien-19, granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin 1β, interleukin 10, and tumor necrosis factor, were identified both at first measurement and across hospitalization (P Conclusions Literature to date demonstrates compelling evidence of increased neutrophils in the circulation and lungs of COVID-19 patients. Importantly, neutrophil quantity and activation correlates with severity of disease. Similarly, our data show that circulating neutrophils in COVID-19 exhibit an activated phenotype with enhanced NETosis and oxidative burst.

Increased plasma mitochondrial DNA concentrations are associated with poor outcomes in multiple critical illnesses, including COVID-19. However, current methods of cell-free mitochondrial DNA quantification in plasma are time-consuming and lack reproducibility. Here, we used next-generation sequencing to characterize the size and genome location of circulating mitochondrial DNA in critically ill subjects with COVID-19 to develop a facile and optimal method of quantification by droplet digital PCR. Sequencing revealed a large percentage of small mitochondrial DNA fragments in plasma with wide variability in coverage by genome location. We identified probes for the mitochondrial DNA genes, cytochrome B and NADH dehydrogenase 1, in regions of relatively high coverage that target small sequences potentially missed by other methods. Serial assessments of absolute mitochondrial DNA concentrations were then determined in plasma from 20 critically ill subjects with COVID-19 without a DNA isolation step. Mitochondrial DNA concentrations on the day of enrollment were increased significantly in patients with moderate or severe acute respiratory distress syndrome (ARDS) compared with those with no or mild ARDS. Comparisons of mitochondrial DNA concentrations over time between patients with no/mild ARDS who survived, patients with moderate/severe ARDS who survived, and nonsurvivors showed the highest concentrations in patients with more severe disease. Absolute mitochondrial DNA quantification by droplet digital PCR is time-efficient and reproducible; thus, we provide a valuable tool and rationale for future studies evaluating mitochondrial DNA as a real-time biomarker to guide clinical decision-making in critically ill subjects with COVID-19.

School readiness is a construct used by educators and policy makers to describe a range of abilities that are beneficial for children transitioning to school. The association of socioeconomic disadvantage with developmental vulnerability when children start school is well established. Parents play a crucial role in supporting children’s transition to school and are acknowledged as their child’s first and foremost teacher. The aim of this study was to explore how parents experience and support their children’s transition to school. This study presents findings from an ethnographic study with 39 parents living in two Australian communities characterised by high area-level socioeconomic disadvantage. The study reports on how parents experience and support their child for transition to school, specifically through their understanding of the concept of ‘readiness’. The role of early childhood services in supporting parents is also explored. Parents conceptualised school readiness as the child’s capacity to function effectively and independently in the hitherto unfamiliar physical and social world of school. Parents considered early childhood services supportive of their children's transition to school, particularly the development of social skills.

tTasmanian early childhood services (ECS) are attempting to improve their engagement with vulnera-ble families and increase the uptake of universal health and education services in this population. Thispaper presents qualitative findings from the [name] Study, focusing on the scope and role of outreachin supporting family engagement in the Tasmanian ECS. Using an ethnographic study design, over 100naturalistic observations were recorded in health and education ECS settings in Tasmanian communitiesbetween April 2017 and February 2018. In addition, 42 ECS providers and 32 parents/guardians withpre-school aged children participated in semi-structured interviews. Interview transcripts and obser-vation field notes were analysed thematically, focusing on family engagement and the role of outreachacross three key universal ECS. Outreach was undertaken by all services involved in this study, but var-ied in practice and scope. Outreach was not directed at specific population groups, but was instigatedin response to an identified need with more vulnerable families to address issues of inequity in serviceaccess. Policies and strategic frameworks within services provided little or no guidance about outreachbeyond procedures for conducting home visits. Attending specialised services alongside parents, a strat-egy adopted by one service, was particularly effective for facilitating connection to services for vulnerablefamilies. The capacity of services to offer outreach was constrained by structured service systems, indi-vidual providers’ skills and capability, resource limitations and lack of clarity with respect to policiesand procedures. Outreach activities are occurring within the universal ECS system in Tasmania, facilitat-ing engagement with vulnerable Tasmanian families and children. Flexible service systems, building theskills and capacities of service providers and clearer policies and procedures would enable services tomore fully embed outreach practices within existing ECS.

Background In Australia, the health and education sectors provide universal early childhood services for the same population of children. Therefore, there is a strong imperative to view service use and outcomes through a cross-sectoral lens to better understand and address the service needs of young children and their families. Objectives To investigate patterns of health and education service use from birth through Kindergarten (age four years), the associations with cumulative risks, and developmental vulnerability in the first year of full-time school (age five years). Methods A retrospective cohort study that used population-wide linkage of health and education administrative data records for 5,440 children with a Tasmanian 2015 Australian Early Development Census (AEDC) record who were born in Tasmania (2008–2010). Results Four service use patterns were identified: Regular (46% of children), Declining (24%); Low (18%); and Selective service use (12%). Regular service use (aOR 0.8, 95% CI 0.7 to 0.9), adjusted for cumulative risks, was associated with decreased odds of developmental vulnerability, compared to the other service use groups. Low (OR 6.1, 95% CI 4.5 to 8.2) and Declining service use (OR 2.5 95% CI 1.9 to 3.4) were more likely for children with the highest levels of cumulative risks. Low and Declining service use, adjusted for cumulative risks were associated with increased odds of developmental vulnerability, compared to the Regular service use group. Conclusion This study provides a whole population view of the differential use of universal services and the complex risk circumstances that influence service use. The association between patterns of multiple risk and service use points to barriers to service use, and the varying level of developmental vulnerability within each service use group draws attention to children who may benefit from higher sustained participation in core health and education services across the whole of early childhood.

The contribution of transcription factors (TFs) and gene regulatory programs in the immune response to COVID-19 and their relationship to disease outcome is not fully understood. Analysis of genome-wide changes in transcription at both promoter-proximal and distal cis-regulatory DNA elements, collectively termed the ’active cistrome,’ offers an unbiased assessment of TF activity identifying key pathways regulated in homeostasis or disease. Here, we profiled the active cistrome from peripheral leukocytes of critically ill COVID-19 patients to identify major regulatory programs and their dynamics during SARS-CoV-2 associated acute respiratory distress syndrome (ARDS). We identified TF motifs that track the severity of COVID- 19 lung injury, disease resolution, and outcome. We used unbiased clustering to reveal distinct cistrome subsets delineating the regulation of pathways, cell types, and the combinatorial activity of TFs. We found critical roles for regulatory networks driven by stimulus and lineage determining TFs, showing that STAT and E2F/MYB regulatory programs targeting myeloid cells are activated in patients with poor disease outcomes and associated with single nucleotide genetic variants implicated in COVID-19 susceptibility. Integration with single-cell RNA-seq found that STAT and E2F/MYB activation converged in specific neutrophils subset found in patients with severe disease. Collectively we demonstrate that cistrome analysis facilitates insight into disease mechanisms and provides an unbiased approach to evaluate global changes in transcription factor activity and stratify patient disease severity.

Objective Using a qualitative design this study aimed to 1) explore the attitudes towards and understanding of osteoarthritis (OA) held by Tasmanian general practitioners (GPs) and orthopaedic surgeons, 2) gain a deeper understanding of conservative and surgical management and 3) identify key barriers and challenges. Design Purposive sampling was used to recruit 17 ​GPs and 10 surgeons from Tasmania, Australia. Semi-structured interviews were audio-recorded, transcribed, coded, and thematically analysed to document understanding of OA, management and treatment decision making. Results GPs and surgeons had a shared understanding of the cause and management of OA which aligned well with evidence-based best practice. Most GPs acknowledged that severity of disease on an X-Ray does not correlate well with symptoms, although some GPs reported always using imaging to support their diagnosis. Conservative management was highly supported by all interviewees, focussing on exercise and/or physiotherapy. Key treatment barriers included managing poor patient understanding of OA, unrealistic expectations for treatment, lack of patient motivation and scepticism towards exercise, and cost and accessibility of conservative treatment options. Surgery was considered a suitable option when conservative management options had been exhausted. Conclusion This study uniquely interviewed GPs and surgeons from the same population, capturing two crucial areas of OA management. Some key barriers to treatment were identified and options for improving treatment include creating opportunities for increased patient education about OA, enhanced accessibility to OA conservative management programs along with improved reimbursement models supporting conservative management as first-line OA treatment.

Background General practice registrars see fewer older patients with chronic disease than established general practitioners (GPs). Furthermore, registrars have fewer opportunities to be involved in continuity of care for older patients. Objective The aim of this study was to explore the experiences of general practice supervisors and registrars when providing in-practice care for older patients with chronic disease. Methods Three focus groups and 21 interviews (18 supervisors, 17 registrars) were conducted, recorded and transcribed. The main themes were derived using thematic analysis. Results Three main themes were reported: context influences registrars’ exposure to older patients; opportunities for continuity of care need ongoing negotiation and communication; registrars are competent – trust and confidence follows. Discussion Developing tailored models of shared patient care that suit different practices and supervisors will require ongoing negotiation and communication. This study confirms the need to enhance exposure for general practice registrars in ways that build on the competence of registrars and the trust in registrars by older patients and supervisors.

Increased awareness of the health disparities faced by lesbian, gay, bisexual, transgender, intersex and queer (LGBTIQ) people has driven the need for LGBTIQ-inclusive medical practices internationally. However, despite bisexual, pansexual and queer women's increased sexual health risks and reduced engagement with health services, there is little qualitative research examining their healthcare experiences. In addition, healthcare practitioners continue to report lack of awareness and competence in inclusive practice, particularly regarding these groups. To address these gaps in the literature and practice, this study draws on 21 qualitative interviews with women and general practitioners, comparing and contrasting their understandings and experiences of inclusive sexual and reproductive healthcare. Findings reveal that women value practitioners who take a non-judgemental approach, use inclusive language and are knowledgeable or willing to self-educate about LGBTIQ issues. Practitioners describe prioritising visual indicators of inclusivity, using inclusive language and embracing professional development. However, women and doctors both identify knowledge gaps among healthcare providers and the need for additional training opportunities to support effective inclusive practice.

Author(s): Han, Claudia; Li, Rick; Hansen, Emily; Bennett, Hunter; Poirion, Olivier; Buchanan, Justin; Challacombe, Jean; Fixsen, Bethany; Trescott, Samantha; Schlachetzki, Johannes CM; Preissl, Sebastian; Wang, Allen; O’Connor, Carolyn; Warden, Anna; Shriram, Shreya; Kim, Roy; Nguyen, Celina; Schafer, Danielle; Ramirez, Gabriela; Anavim, Samuel; Johnson, Avalon; Sajti, Eniko; Gupta, Mihir; Levy, Michael; Ben-Haim, Sharona; Gonda, David; Laurent, Louise; Glass, Christopher; Coufal, Nicole | Abstract: The fetal period is a critical time for brain development, characterized by neurogenesis, neural migration, and synaptogenesis 1-3 . Microglia, the tissue resident macrophages of the brain, are observed as early as the fourth week of gestation 4 and are thought to engage in a variety of processes essential for brain development and homeostasis 5-11 . Conversely, microglia phenotypes are highly regulated by the brain environment 12-14 . Mechanisms by which human brain development influences the maturation of microglia and microglia potential contribution to neurodevelopmental disorders remain poorly understood. Here, we performed transcriptomic analysis of human fetal and postnatal microglia and corresponding cortical tissue to define age-specific brain environmental factors that may drive microglia phenotypes. Comparative analysis of open chromatin profiles using bulk and single-cell methods in conjunction with a new computational approach that integrates epigenomic and single-cell RNA-seq data allowed decoding of cellular heterogeneity with inference of subtype- and development stage-specific transcriptional regulators. Interrogation of in vivo and in vitro iPSC-derived microglia models provides evidence for roles of putative instructive signals and downstream gene regulatory networks which establish human-specific fetal and postnatal microglia gene expression programs and potentially contribute to neurodevelopmental disorders.

Introduction. The pathophysiology of infection with SARS-CoV-2 involves the lower airways and host-launched aggressive inflammatory responses leading to exacerbated lung damage in these vital tissues. Early clinical studies found that COVID-19 patients have higher levels of neutrophils in the circulation. Neutrophils are the most abundant leukocyte in circulation and are known to be highly proinflammatory due to production of neutrophil extracellular traps (NETosis). NETs are web-like chromatin structures coated with histones and proteases that both capture and kill invading pathogens. However, while being an effective countermeasure towards foreign microbes, this process also causes undesirable damage in host tissues. Therefore, we sought to characterize NETosis in circulating neutrophils from COVID-19 patients to determine whether this immunological response might be exacerbating or driving the disease state in COVID-19, rather than mitigating the virus. Methods.Blood was drawn daily from critically ill COVID-19 patients (n=16) after consent was obtained. Healthy controls (n=13) were screened for COVID-19 and gave blood once a week. Blood was drawn into lithium heparin tubes (BD Vacutainer). Neutrophils were isolated using PolymorphprepTM(PROGEN) per manufacturer's instructions. Cells were resuspended at 2x106 cells/ml for functional assays. Neutrophils were stimulated with increasing concentrations of PMA (Phorbol 12-myristate 13-acetate) of 2.5nM, 25nM and 250nM to stimulate NETosis via the canonical pathway, and nigericin at 15uM for the non-canonical pathway. NETosis was quantified using the Quant-iT™ PicoGreen™ dsDNA Assay Kit (Invitrogen) and by NET visualization via myeloperoxidase and nuclear staining (using Polyclonal Rabbit Anti-Human Myeloperoxidase by Dako and Hoescht stain by Invitrogen). Results.Functional NETosis assays of circulating neutrophils from COVID-19 patients demonstrate overall increased NETosis determined by increased release of dsDNA. This enhanced NETosis occurred at baseline and after stimulation with PMA when compared to healthy controls (Figure 1A, p

BackgroundIncreased inflammation is a hallmark of COVID-19, with pulmonary and systemic inflammation identified in multiple cohorts of patients. Definitive cellular and molecular pathways driving severe forms of this disease remain uncertain. Neutrophils, the most numerous leukocytes in blood circulation, can contribute to immunopathology in infections, inflammatory diseases and acute respiratory distress syndrome (ARDS), a primary cause of morbidity and mortality in COVID-19. Changes in multiple neutrophil functions and circulating cytokine levels over time during COVID-19 may help define disease severity and guide care and decision making.MethodsBlood was obtained serially from critically ill COVID-19 patients for 11 days. Neutrophil oxidative burst, neutrophil extracellular trap formation (NETosis), phagocytosis and cytokine levels were assessed ex vivo. Lung tissue was obtained immediately post-mortem for immunostaining.ResultsElevations in neutrophil-associated cytokines IL-8 and IL-6, and general inflammatory cytokines IP-10, GM-CSF, IL-1b, IL-10 and TNF, were identified in COVID-19 plasma both at the first measurement and at multiple timepoints across hospitalization (p < 0.0001). Neutrophils had exaggerated oxidative burst (p < 0.0001), NETosis (p < 0.0001) and phagocytosis (p < 0.0001) relative to controls. Increased NETosis correlated with both leukocytosis and neutrophilia. Neutrophils and NETs were identified within airways and alveoli in the lung parenchyma of 40% of SARS-CoV-2 infected lungs. While elevations in IL-8 and ANC correlated to COVID-19 disease severity, plasma IL-8 levels alone correlated with death.ConclusionsCirculating neutrophils in COVID-19 exhibit an activated phenotype with increased oxidative burst, NETosis and phagocytosis. Readily accessible and dynamic, plasma IL-8 and circulating neutrophil function may be potential COVID-19 disease biomarkers.

International audience; We present the largest exome sequencing study of autism spectrum disorder (ASD) to date (n = 35,584 total samples, 11,986 with ASD). Using an enhanced analytical framework to integrate de novo and case-control rare variation, we identify 102 risk genes at a false discovery rate of 0.1 or less. Of these genes, 49 show higher frequencies of disruptive de novo variants in individuals ascertained to have severe neurodevelopmental delay, whereas 53 show higher frequencies in individuals ascertained to have ASD; comparing ASD cases with mutations in these groups reveals phenotypic differences. Expressed early in brain development, most risk genes have roles in regulation of gene expression or neuronal communication (i.e., mutations effect neurodevelopmental and neurophysiological changes), and 13 fall within loci recurrently hit by copy number variants. In cells from the human cortex, expression of risk genes is enriched in excitatory and inhibitory neuronal lineages, consistent with multiple paths to an excitatory-inhibitory imbalance underlying ASD.