LINE-1 retrotransposons contribute to mouse PV interneuron development.

Retrotransposons are mobile DNA sequences duplicated via transcription and reverse transcription of an RNA intermediate. Cis-regulatory elements encoded by retrotransposons can also promote the transcription of adjacent genes. Somatic LINE-1 (L1) retrotransposon insertions have been detected in mammalian neurons. It is, however, unclear whether L1 sequences are mobile in only some neuronal lineages or therein promote neurodevelopmental gene expression. Here we report programmed L1 activation by SOX6, a transcription factor critical for parvalbumin (PV) interneuron development. Mouse PV interneurons permit L1 mobilization in vitro and in vivo, harbor unmethylated L1 promoters and express full-length L1 mRNAs and proteins. Using nanopore long-read sequencing, we identify unmethylated L1s proximal to PV interneuron genes, including a novel L1 promoter-driven Caps2 transcript isoform that enhances neuron morphological complexity in vitro. These data highlight the contribution made by L1 cis-regulatory elements to PV interneuron development and transcriptome diversity, uncovered due to L1 mobility in this milieu.

Every repeat is unique: Exploring the genomic impact of human L1 retrotransposons at locus-specific resolution.

Fully understanding the impact of the human retrotransposon L1 requires that each of ∼500,000 L1 copies be evaluated as a potentially unique genomic entity. In this issue of Cell Genomics, Lanciano et al.1 strive toward this goal, illuminating the reciprocal regulatory influence between individual L1s and their genomic integration sites.

Locus-resolution analysis of L1 regulation and retrotransposition potential in mouse embryonic development.

Mice harbor ∼2800 intact copies of the retrotransposon Long Interspersed Element 1 (L1). The in vivo retrotransposition capacity of an L1 copy is defined by both its sequence integrity and epigenetic status, including DNA methylation of the monomeric units constituting young mouse L1 promoters. Locus-specific L1 methylation dynamics during development may therefore elucidate and explain spatiotemporal niches of endogenous retrotransposition but remain unresolved. Here, we interrogate the retrotransposition efficiency and epigenetic fate of source (donor) L1s, identified as mobile in vivo. We show that promoter monomer loss consistently attenuates the relative retrotransposition potential of their offspring (daughter) L1 insertions. We also observe that most donor/daughter L1 pairs are efficiently methylated upon differentiation in vivo and in vitro. We use Oxford Nanopore Technologies (ONT) long-read sequencing to resolve L1 methylation genome-wide and at individual L1 loci, revealing a distinctive "smile" pattern in methylation levels across the L1 promoter region. Using Pacific Biosciences (PacBio) SMRT sequencing of L1 5' RACE products, we then examine DNA methylation dynamics at the mouse L1 promoter in parallel with transcription start site (TSS) distribution at locus-specific resolution. Together, our results offer a novel perspective on the interplay between epigenetic repression, L1 evolution, and genome stability.

An international expansion in voluntary euthanasia/assisted dying: The implications for nursing.

AIM: To provide an update on the international position of assisted dying legislation and its implications for nursing policy. BACKGROUND: Assisted dying legislation has been introduced in health systems in Europe, North America and Australasia (Australia and New Zealand). Despite contributions in research literature, this remains focussed on medical practitioners, with limited acknowledgement of the need for policy development in nursing. DISCUSSION: There is a need for critical evaluation of this contemporary issue and the significance for nursing practice of the lack of unified nursing perspective and robust policy guidance is identified. An overview of the existing role of voluntary euthanasia/assisted dying with recent developments is provided and the resulting concerns for nurses regarding the scope of practice, role confusion and conflicting professional values is considered. CONCLUSION: Despite a long history of assisted dying and the continued expansion of these practices, limited and highly variable nursing policy highlights the lack of clear guidance available to nurses. The growing inclusion of nurse practitioners and recognition of registered nurse involvement in the care of individuals on an assisted dying pathway merits clearer support from regulatory authorities and professional organisations. IMPLICATIONS FOR NURSING PRACTICE: Nurses are the most likely health professionals to be approached with questions regarding assisted dying; they are intimately involved in patient experiences and need to understand what is possible, expected and legal in terms of their scope of practice regarding assisted dying. IMPLICATIONS FOR NURSING POLICY: Nursing regulators and professional representatives need to provide clear policy statements and guidance identifying the nursing role and recognising where protections and support are necessary.

Therapeutic inefficacy of protocol driven intravenous unfractionated heparin infusion in the current era.

Unfractionated heparin (UFH) is commonly used for several life-threatening conditions requiring anticoagulant therapy but failure to reach therapeutic levels in 24 h can be associated with adverse outcomes. Use of low molecular weight heparin (LMWH) may provide an alternative while providing superior outcomes as compared to UFH. We studied 100 patients who underwent UFH therapy for >24 h and found that theoretically 80 % were eligible for LMWH therapy. Only 29 % and 40 % of the total aPTT draws were in the therapeutic window within the first 24 h and at 25-48 h respectively. This study reports that a vast majority of patients remain outside of therapeutic aPTT within first 24-48 h when anticoagulated with UFH. With high eligibility for LMWH therapy, its substitution can potentially lead to better patient outcomes, higher levels of therapeutic efficacy, and decrease in hospital resources.

Retrotransposon instability dominates the acquired mutation landscape of mouse induced pluripotent stem cells.

Induced pluripotent stem cells (iPSCs) can in principle differentiate into any cell of the body, and have revolutionized biomedical research and regenerative medicine. Unlike their human counterparts, mouse iPSCs (miPSCs) are reported to silence transposable elements and prevent transposable element-mediated mutagenesis. Here we apply short-read or Oxford Nanopore Technologies long-read genome sequencing to 38 bulk miPSC lines reprogrammed from 10 parental cell types, and 18 single-cell miPSC clones. While single nucleotide variants and structural variants restricted to miPSCs are rare, we find 83 de novo transposable element insertions, including examples intronic to Brca1 and Dmd. LINE-1 retrotransposons are profoundly hypomethylated in miPSCs, beyond other transposable elements and the genome overall, and harbor alternative protein-coding gene promoters. We show that treatment with the LINE-1 inhibitor lamivudine does not hinder reprogramming and efficiently blocks endogenous retrotransposition, as detected by long-read genome sequencing. These experiments reveal the complete spectrum and potential significance of mutations acquired by miPSCs.

Challenging the culture of Emergency Department violence and aggression.

AIMS: To examine reported levels of violence and aggression within a tertiary level emergency department in New Zealand, and to compare incident reporting within a dedicated yearly audit period to standard organisational reporting procedures. METHOD: A prospective, longitudinal cohort study involving repeated yearly audits of violence and aggression reported by emergency department staff from 2014-2020. RESULTS: Episodes of violence and aggression were reported at high levels during audit months compared to standard reporting, suggesting current systems do not accurately reflect the presence of violence and aggression. Levels of reported violence and aggression remained relatively static over a seven-year period, despite increasing emergency department attendances. Most events reported involved verbal abuse from patients, and occurred on weekend and night shifts. A number of potentially contributing factors were identified. CONCLUSIONS: Persistently higher levels of violence and aggression were reported during the targeted audit months, while reporting via the organisation's formal system during the intervening months remained at low levels. Further research is essential to monitor trends, assess the effectiveness of interventions to improve reporting, modify factors contributing to violence and aggression, and to address the impact on staff and bystanders affected in emergency departments.

Somatic retrotransposition in the developing rhesus macaque brain.

The retrotransposon LINE-1 (L1) is central to the recent evolutionary history of the human genome and continues to drive genetic diversity and germline pathogenesis. However, the spatiotemporal extent and biological significance of somatic L1 activity are poorly defined and are virtually unexplored in other primates. From a single L1 lineage active at the divergence of apes and Old World monkeys, successive L1 subfamilies have emerged in each descendant primate germline. As revealed by case studies, the presently active human L1 subfamily can also mobilize during embryonic and brain development in vivo. It is unknown whether nonhuman primate L1s can similarly generate somatic insertions in the brain. Here we applied approximately 40× single-cell whole-genome sequencing (scWGS), as well as retrotransposon capture sequencing (RC-seq), to 20 hippocampal neurons from two rhesus macaques (Macaca mulatta). In one animal, we detected and PCR-validated a somatic L1 insertion that generated target site duplications, carried a short 5' transduction, and was present in ∼7% of hippocampal neurons but absent from cerebellum and nonbrain tissues. The corresponding donor L1 allele was exceptionally mobile in vitro and was embedded in PRDM4, a gene expressed throughout development and in neural stem cells. Nanopore long-read methylome and RNA-seq transcriptome analyses indicated young retrotransposon subfamily activation in the early embryo, followed by repression in adult tissues. These data highlight endogenous macaque L1 retrotransposition potential, provide prototypical evidence of L1-mediated somatic mosaicism in a nonhuman primate, and allude to L1 mobility in the brain over the past 30 million years of human evolution.

District nursing and family/whanau assessment practices: A New Zealand study.

AIM: District Nurses apply specialized nursing knowledge and assessment skills to provide care in New Zealand communities. This study aimed to identify whether District Nurse's (both Registered and supervised Enrolled Nurse's) had knowledge of, and used the 15-Minute Interview tool, including Ecomaps/Genograms, and if not, what they saw as enablers or barriers to doing so. DESIGN: Participatory action research was used, following the phases of look, think and act. METHODS: Two pre-intervention focus groups occurred, two education sessions which introduced the 15-Minute Interview and four postintervention interviews which explored the use of the tools and their potential use in the future. RESULTS: District Nurses demonstrated working with families, and the selection of when and where to apply the 15-Minute Interview.

Intensive care nurse-family engagement from a global perspective: A qualitative multi-site exploration.

BACKGROUND: Critical illness is distressing for families, and often results in negative effects on family health that influence a family's ability to support their critically ill family member. Although recent attention has been directed at improving care and outcomes for families of critically ill patients, the manner in which nurses engage with families is not fully understood. OBJECTIVES: To describe nurses' perceptions and practices of family engagement in adult intensive care units from a global perspective. DESIGN: A qualitative-descriptive multi-site design using content analysis. SETTINGS: The study was conducted in 26 intensive care units of 12 urban, metropolitan, academic medical centers in ten countries, spanning five continents. PARTICIPANTS: A total of 65 registered nurses (77% women, age of M = 39.5, SD = 11.4 years) participated. Most held intensive care certification (72%) and had worked on average 10 (SD = 9.6) years in the ICU. METHODS: Semi-structured, individual interviews (M = 38.4 min, SD = 12.0) were held with ICU nurses at the hospital (94%) or their home using an interview guide. Qualitative interview data were analysed using inductive content analysis. RESULTS: We found that nurse-family engagement was an ebb and flow of relational power that needed to be carefully negotiated and balanced, with nurses holding and often exerting more power than families. Constant fluctuations in nurses' practices of engagement occurred in day-to-day practice from shift-to-shift and from nurse-to-nurse. Family engagement was dependent on individual nurses' attitudes and perceptions of family, the patient's condition, and workload. Lastly, family engagement was shaped by the ICU context, with team culture, collaborative relationships, unit structures and organizational resources either enabling or limiting nurses' ability to engage with families. CONCLUSIONS: This global study provides an in-depth understanding of the way nurses engage with families in ICU and reflects many different cultures and health systems. We found that nurse-family engagement was marked by a shifting, yet often unequal power distribution in the nurse-family relationship, inconsistent nurse engagement practices, both of which resulted in variable family engagement in intensive care. Our research contributes a detailed description of engagement as practiced in the everyday delivery of health care. A more concentrated team effort, based on a shared culture and defined framework of family care is needed to ensure that families of critically ill persons are fully engaged in all aspects of intensive care.

Mental health presentations to Christchurch Hospital Emergency Department during COVID-19 lockdown.

OBJECTIVE: To describe mental health presentations to a tertiary ED in New Zealand during a national COVID-19 lockdown. METHODS: A retrospective, comparative cohort study in Christchurch Hospital, New Zealand. RESULTS: There was a 3510 (37%)-patient decrease in all presentations to Christchurch Hospital ED during the 5-week COVID-19 lockdown period from 26 March 2020 to 28 April 2020, compared to a 111 (1.2%)-patient decrease in the same time period in the previous year (P < 0.00001). There is usually a seasonal reduction in mental health attendances at this time of year compared to the weeks before. In 2019, there was a 49 (9.8%)-patient reduction in mental health presentations, whereas in 2020 there was a 193 (34%)-patient reduction (P < 0.001). In 2020, the proportion of mental health attendances compared to all ED attendances during the 5-week lockdown period was similar to the 5-week pre-lockdown period (564/9460 vs 371/5950, P = 0.48). The proportion of mental health patients presenting due to overdose increased by 6.5% (158/564 vs 128/371, P = 0.035); those due to self-harm increased by 3.5% (35/564 vs 36/371, P = 0.049). The proportion of mental health presentations due to anxiety, depression and other non-self-harm/overdose complaints decreased by 10% (371/564 vs 207/371, P = 0.002). The proportion of overdoses of paracetamol and ibuprofen increased by 13.4% during lockdown (22/158 vs 35/128, P = 0.005). CONCLUSIONS: During the COVID-19 lockdown, both overall ED presentations as well as mental health-related presentations decreased. There was a relative increase in overdoses and self-harm, particularly involving paracetamol and ibuprofen.

Nanopore Sequencing Enables Comprehensive Transposable Element Epigenomic Profiling.

Transposable elements (TEs) drive genome evolution and are a notable source of pathogenesis, including cancer. While CpG methylation regulates TE activity, the locus-specific methylation landscape of mobile human TEs has to date proven largely inaccessible. Here, we apply new computational tools and long-read nanopore sequencing to directly infer CpG methylation of novel and extant TE insertions in hippocampus, heart, and liver, as well as paired tumor and non-tumor liver. As opposed to an indiscriminate stochastic process, we find pronounced demethylation of young long interspersed element 1 (LINE-1) retrotransposons in cancer, often distinct to the adjacent genome and other TEs. SINE-VNTR-Alu (SVA) retrotransposons, including their internal tandem repeat-associated CpG island, are near-universally methylated. We encounter allele-specific TE methylation and demethylation of aberrantly expressed young LINE-1s in normal tissues. Finally, we recover the complete sequences of tumor-specific LINE-1 insertions and their retrotransposition hallmarks, demonstrating how long-read sequencing can simultaneously survey the epigenome and detect somatic TE mobilization.

The Family Meal Model: Influences on Family Mealtime Participation.

The family meal affords benefits such as positive nutritional habits, trust building, connecting, parent modeling, and teaching. During the school-aged years, families can support children's development of health behaviors and family routines. This interdisciplinary study examined families' experiences of mealtimes and the factors that support or hinder mealtime participation. Grounded theory methods guided data collection and analysis. Semi-structured interviews were conducted with 21 families with at least one school-aged child (n = 68). Interviews were audio recorded and transcribed. Researchers coded and categorized data, identified themes, and generated a model. Families defined family mealtime as all family members being together at the kitchen or dining room table eating a meal. Families enjoyed being together, conversing, and connecting through family mealtime participation. The Family Meal Model proposes relationships between factors that support or hinder a family's mealtime participation. Discussion includes support for a broadened role of occupational therapy in promoting family occupation through family meals.

Management of the aggressive emergency department patient: non-pharmacological perspectives and evidence base.

INTRODUCTION: Aggression in the Emergency Department (ED) remains an ongoing issue, described as reaching epidemic proportions, with an impact on staff recruitment, retention, and ability to provide quality care. Most literature has focused on the definition (or lack of) core concepts, efforts to quantify the phenomenon or provide an epidemiological profile. Relatively little offers evidence-based interventions or evaluations of the same. AIM: To identify the range of suggested practices and the evidence base for currently recommended actions relating to the management of the aggressive Emergency Department patient. METHODS: A meta-synthesis of existing reviews of violence and aggression in the acute health-care setting, including management of the aggressive patient, was undertaken. This provided the context for critical consideration of the management of this patient group in the ED and implications for clinical practice. RESULTS: An initial outline of issues was followed by a systematic search and 15 reviews were further assessed. Commonly identified interventions are grouped around educational, interpersonal, environmental, and physical responses. These actions can be focused in terms of overall responses to the wider issues of violence and aggression, targeted at the pre-event, event, or post-event phase in terms of strategies; however, there is a very limited evidence base to show the effectiveness of strategies suggested. CLINICAL IMPLICATIONS: The lack of evidence-based intervention strategies leaves clinicians in a difficult situation, often enacting practices based on anecdote rather than evidence. Local solutions to local problems are occurring in a pragmatic manner, but there needs to be clarification and integration of workable processes for evaluating and disseminating best practice. CONCLUSION: There is limited evidence reporting on interventional studies, in addition to identification of the need for high quality longitudinal and evaluation studies to determine the efficacy of those responses that have been identified.

LINE-1 Evasion of Epigenetic Repression in Humans.

Epigenetic silencing defends against LINE-1 (L1) retrotransposition in mammalian cells. However, the mechanisms that repress young L1 families and how L1 escapes to cause somatic genome mosaicism in the brain remain unclear. Here we report that a conserved Yin Yang 1 (YY1) transcription factor binding site mediates L1 promoter DNA methylation in pluripotent and differentiated cells. By analyzing 24 hippocampal neurons with three distinct single-cell genomic approaches, we characterized and validated a somatic L1 insertion bearing a 3' transduction. The source (donor) L1 for this insertion was slightly 5' truncated, lacked the YY1 binding site, and was highly mobile when tested in vitro. Locus-specific bisulfite sequencing revealed that the donor L1 and other young L1s with mutated YY1 binding sites were hypomethylated in embryonic stem cells, during neurodifferentiation, and in liver and brain tissue. These results explain how L1 can evade repression and retrotranspose in the human body.

Dynamic Methylation of an L1 Transduction Family during Reprogramming and Neurodifferentiation.

The retrotransposon LINE-1 (L1) is a significant source of endogenous mutagenesis in humans. In each individual genome, a few retrotransposition-competent L1s (RC-L1s) can generate new heritable L1 insertions in the early embryo, primordial germ line, and germ cells. L1 retrotransposition can also occur in the neuronal lineage and cause somatic mosaicism. Although DNA methylation mediates L1 promoter repression, the temporal pattern of methylation applied to individual RC-L1s during neurogenesis is unclear. Here, we identified a de novo L1 insertion in a human induced pluripotent stem cell (hiPSC) line via retrotransposon capture sequencing (RC-seq). The L1 insertion was full-length and carried 5' and 3' transductions. The corresponding donor RC-L1 was part of a large and recently active L1 transduction family and was highly mobile in a cultured-cell L1 retrotransposition reporter assay. Notably, we observed distinct and dynamic DNA methylation profiles for the de novo L1 and members of its extended transduction family during neuronal differentiation. These experiments reveal how a de novo L1 insertion in a pluripotent stem cell is rapidly recognized and repressed, albeit incompletely, by the host genome during neurodifferentiation, while retaining potential for further retrotransposition.

Psychological health and resilience: the impact of significant earthquake events on tertiary level professional students. A cross-sectional study.

BACKGROUND: The effect of natural disasters internationally is linked to intensity and duration and the impact of these events for tertiary level professional students is not clearly understood. Following a 7.1 magnitude earthquake in New Zealand in 2010 (with aftershocks lasting 27 months) a number of tertiary nursing students experienced significant disruption to their studies. AIM: To compare the psychological health, resilience and the impact on learning for three cohorts of students engaged in tertiary nursing education during this time. METHOD: A cross-sectional survey design and convenience sampling was used for three cohorts of learners. An online survey was completed (n = 290) and included: Depression Anxiety and Stress Scale; PTSD Checklist; Work and Social Adjustment Scale; Connor-Davidson Resilience Scale. RESULTS: Statistically significant differences were found across the psychometric scales with regard to relationship status. Whilst an increase in self-reported physical and mental health issues prior to and following the earthquakes were noted, mitigating factors were also identified. CONCLUSIONS: In order to support psychological health amongst nursing students, tertiary education systems need to plan for sustainable learning. The importance of facilitating future orientation within organisations is necessary to develop resilience amongst staff and students, which, in turn, will enable on-going education during significant disaster events.

L1 Retrotransposon Heterogeneity in Ovarian Tumor Cell Evolution.

LINE-1 (L1) retrotransposons are a source of insertional mutagenesis in tumor cells. However, the clinical significance of L1 mobilization during tumorigenesis remains unclear. Here, we applied retrotransposon capture sequencing (RC-seq) to multiple single-cell clones isolated from five ovarian cancer cell lines and HeLa cells and detected endogenous L1 retrotransposition in vitro. We then applied RC-seq to ovarian tumor and matched blood samples from 19 patients and identified 88 tumor-specific L1 insertions. In one tumor, an intronic de novo L1 insertion supplied a novel cis-enhancer to the putative chemoresistance gene STC1. Notably, the tumor subclone carrying the STC1 L1 mutation increased in prevalence after chemotherapy, further increasing STC1 expression. We also identified hypomethylated donor L1s responsible for new L1 insertions in tumors and cultivated cancer cells. These congruent in vitro and in vivo results highlight L1 insertional mutagenesis as a common component of ovarian tumorigenesis and cancer genome heterogeneity.