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BackgroundApolipoprotein E ε4 (APOE4) is the strongest genetic risk factor for late-onset Alzheimer's disease (LOAD). A recent case report identified a rare variant in APOE, APOE3-R136S (Christchurch), proposed to confer resistance to autosomal dominant Alzheimer's Disease (AD). However, it remains unclear whether and how this variant exerts its protective effects.MethodsWe introduced the R136S variant into mouse Apoe (ApoeCh) and investigated its effect on the development of AD-related pathology using the 5xFAD model of amyloidosis and the PS19 model of tauopathy. We used immunohistochemical and biochemical analysis along with single-cell spatial omics and bulk proteomics to explore the impact of the ApoeCh variant on AD pathological development and the brain's response to plaques and tau.ResultsIn 5xFAD mice, ApoeCh enhances a Disease-Associated Microglia (DAM) phenotype in microglia surrounding plaques, and reduces plaque load, dystrophic neurites, and plasma neurofilament light chain. By contrast, in PS19 mice, ApoeCh suppresses the microglial and astrocytic responses to tau-laden neurons and does not reduce tau accumulation or phosphorylation, but partially rescues tau-induced synaptic and myelin loss. We compared how microglia responses differ between the two mouse models to elucidate the distinct DAM signatures induced by ApoeCh. We identified upregulation of antigen presentation-related genes in the DAM response in a PS19 compared to a 5xFAD background, suggesting a differential response to amyloid versus tau pathology that is modulated by the presence of ApoeCh. Bulk proteomics show upregulated mitochondrial protein abundance with ApoeCh in 5xFAD mice, but reductions in mitochondrial and translation associated proteins in PS19 mice.ConclusionsThese findings highlight the ability of the ApoeCh variant to modulate microglial responses based on the type of pathology, enhancing DAM reactivity in amyloid models and dampening neuroinflammation to promote protection in tau models. This suggests that the Christchurch variant's protective effects likely involve multiple mechanisms, including changes in receptor binding and microglial programming.

Abstract Background Apolipoprotein E ε4 (APOE4) is the strongest genetic risk factor for late-onset Alzheimer’s disease (LOAD). A recent case report identified a rare variant in APOE, APOE3-R136S (Christchurch), proposed to confer resistance to autosomal dominant Alzheimer’s Disease (AD). However, it remains unclear whether and how this variant exerts its protective effects. Methods We introduced the R136S variant into mouse Apoe (ApoeCh) and investigated its effect on the development of AD-related pathology using the 5xFAD model of amyloidosis and the PS19 model of tauopathy. We used immunohistochemical and biochemical analysis along with single-cell spatial omics and bulk proteomics to explore the impact of the ApoeCh variant on AD pathological development and the brain’s response to plaques and tau. Results In 5xFAD mice, ApoeCh enhances a Disease-Associated Microglia (DAM) phenotype in microglia surrounding plaques, and reduces plaque load, dystrophic neurites, and plasma neurofilament light chain. By contrast, in PS19 mice, ApoeCh suppresses the microglial and astrocytic responses to tau-laden neurons and does not reduce tau accumulation or phosphorylation, but partially rescues tau-induced synaptic and myelin loss. We compared how microglia responses differ between the two mouse models to elucidate the distinct DAM signatures induced by ApoeCh. We identified upregulation of antigen presentation-related genes in the DAM response in a PS19 compared to a 5xFAD background, suggesting a differential response to amyloid versus tau pathology that is modulated by the presence of ApoeCh. Bulk proteomics show upregulated mitochondrial protein abundance with ApoeCh in 5xFAD mice, but reductions in mitochondrial and translation associated proteins in PS19 mice. Conclusions These findings highlight the ability of the ApoeCh variant to modulate microglial responses based on the type of pathology, enhancing DAM reactivity in amyloid models and dampening neuroinflammation to promote protection in tau models. This suggests that the Christchurch variant's protective effects likely involve multiple mechanisms, including changes in receptor binding and microglial programming. Graphical Abstract

BackgroundLimited information is available for patients with breast cancer (BC) and coronavirus disease 2019 (COVID-19), especially among underrepresented racial/ethnic populations.MethodsThis is a COVID-19 and Cancer Consortium (CCC19) registry-based retrospective cohort study of females with active or history of BC and laboratory-confirmed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection diagnosed between March 2020 and June 2021 in the US. Primary outcome was COVID-19 severity measured on a five-level ordinal scale, including none of the following complications, hospitalization, intensive care unit admission, mechanical ventilation, and all-cause mortality. Multivariable ordinal logistic regression model identified characteristics associated with COVID-19 severity.Results1383 female patient records with BC and COVID-19 were included in the analysis, the median age was 61 years, and median follow-up was 90 days. Multivariable analysis revealed higher odds of COVID-19 severity for older age (aOR per decade, 1.48 [95% CI, 1.32-1.67]); Black patients (aOR 1.74; 95 CI 1.24-2.45), Asian Americans and Pacific Islander patients (aOR 3.40; 95 CI 1.70-6.79) and Other (aOR 2.97; 95 CI 1.71-5.17) racial/ethnic groups; worse ECOG performance status (ECOG PS ≥2: aOR, 7.78 [95% CI, 4.83-12.5]); pre-existing cardiovascular (aOR, 2.26 [95% CI, 1.63-3.15])/pulmonary comorbidities (aOR, 1.65 [95% CI, 1.20-2.29]); diabetes mellitus (aOR, 2.25 [95% CI, 1.66-3.04]); and active and progressing cancer (aOR, 12.5 [95% CI, 6.89-22.6]). Hispanic ethnicity, timing, and type of anti-cancer therapy modalities were not significantly associated with worse COVID-19 outcomes. The total all-cause mortality and hospitalization rate for the entire cohort was 9% and 37%, respectively however, it varied according to the BC disease status.ConclusionsUsing one of the largest registries on cancer and COVID-19, we identified patient and BC-related factors associated with worse COVID-19 outcomes. After adjusting for baseline characteristics, underrepresented racial/ethnic patients experienced worse outcomes compared to non-Hispanic White patients.FundingThis study was partly supported by National Cancer Institute grant number P30 CA068485 to Tianyi Sun, Sanjay Mishra, Benjamin French, Jeremy L Warner; P30-CA046592 to Christopher R Friese; P30 CA023100 for Rana R McKay; P30-CA054174 for Pankil K Shah and Dimpy P Shah; KL2 TR002646 for Pankil Shah and the American Cancer Society and Hope Foundation for Cancer Research (MRSG-16-152-01-CCE) and P30-CA054174 for Dimpy P Shah. REDCap is developed and supported by Vanderbilt Institute for Clinical and Translational Research grant support (UL1 TR000445 from NCATS/NIH). The funding sources had no role in the writing of the manuscript or the decision to submit it for publication.Clinical trial numberCCC19 registry is registered on ClinicalTrials.gov, NCT04354701.

Microglia are the primary resident myeloid cells of the brain responsible for maintaining homeostasis and protecting the central nervous system (CNS) from damage and infection. Monocytes and monocyte-derived macrophages arising from the periphery have also been implicated in CNS pathologies, however, distinguishing between different myeloid cell populations in the CNS has been difficult. Here, we set out to develop a reliable histological marker that can assess distinct myeloid cell heterogeneity and functional contributions, particularly in the context of disease and/or neuroinflammation. scRNAseq from brains of mice infected with the neurotropic JHM strain of the mouse hepatitis virus (JHMV), a mouse coronavirus, revealed that Lgals3 is highly upregulated in monocyte and macrophage populations, but not in microglia. Subsequent immunostaining for galectin-3 (encoded by Lgals3), also referred to as MAC2, highlighted the high expression levels of MAC2 protein in infiltrating myeloid cells in JHMV-infected and bone marrow (BM) chimeric mice, in stark contrast to microglia, which expressed little to no staining in these models. Expression of MAC2 was found even 6-10 months following BM-derived cell infiltration into the CNS. We also demonstrate that MAC2 is not a specific label for plaque-associated microglia in the 5xFAD mouse model, but only appears in a distinct subset of these cells in the presence of JHMV infection or during aging. Our data suggest that MAC2 can serve as a reliable and long-lasting histological marker for monocyte/macrophages in the brain, identifying an accessible approach to distinguishing resident microglia from infiltrating cells in the CNS under certain conditions.

Unlike SARS-CoV-1 and MERS-CoV, infection with SARS-CoV-2, the viral pathogen responsible for COVID-19, is often associated with neurologic symptoms that range from mild to severe, yet increasing evidence argues the virus does not exhibit extensive neuroinvasive properties. We demonstrate SARS-CoV-2 can infect and replicate in human iPSC-derived neurons and that infection shows limited antiviral and inflammatory responses but increased activation of EIF2 signaling following infection as determined by RNA sequencing. Intranasal infection of K18 human ACE2 transgenic mice (K18-hACE2) with SARS-CoV-2 resulted in lung pathology associated with viral replication and immune cell infiltration. In addition, ∼50% of infected mice exhibited CNS infection characterized by wide-spread viral replication in neurons accompanied by increased expression of chemokine (Cxcl9, Cxcl10, Ccl2, Ccl5 and Ccl19) and cytokine (Ifn-λ and Tnf-α) transcripts associated with microgliosis and a neuroinflammatory response consisting primarily of monocytes/macrophages. Microglia depletion via administration of colony-stimulating factor 1 receptor inhibitor, PLX5622, in SARS-CoV-2 infected mice did not affect survival or viral replication but did result in dampened expression of proinflammatory cytokine/chemokine transcripts and a reduction in monocyte/macrophage infiltration. These results argue that microglia are dispensable in terms of controlling SARS-CoV-2 replication in in the K18-hACE2 model but do contribute to an inflammatory response through expression of pro-inflammatory genes. Collectively, these findings contribute to previous work demonstrating the ability of SARS-CoV-2 to infect neurons as well as emphasizing the potential use of the K18-hACE2 model to study immunological and neuropathological aspects related to SARS-CoV-2-induced neurologic disease. IMPORTANCE Understanding the immunological mechanisms contributing to both host defense and disease following viral infection of the CNS is of critical importance given the increasing number of viruses that are capable of infecting and replicating within the nervous system. With this in mind, the present study was undertaken to evaluate the role of microglia in aiding in host defense following experimental infection of the central nervous system (CNS) of K18-hACE2 with SARS-CoV-2, the causative agent of COVID-19. Neurologic symptoms that range in severity are common in COVID-19 patients and understanding immune responses that contribute to restricting neurologic disease can provide important insight into better understanding consequences associated with SARS-CoV-2 infection of the CNS.

Microglia shape the synaptic environment in health and disease, but synapses do not exist in a vacuum. Instead, pre- and postsynaptic terminals are surrounded by extracellular matrix (ECM), which together with glia comprise the four elements of the contemporary tetrapartite synapse model. While research in this area is still just beginning, accumulating evidence points toward a novel role for microglia in regulating the ECM during normal brain homeostasis, and such processes may, in turn, become dysfunctional in disease. As it relates to synapses, microglia are reported to modify the perisynaptic matrix, which is the diffuse matrix that surrounds dendritic and axonal terminals, as well as perineuronal nets (PNNs), specialized reticular formations of compact ECM that enwrap neuronal subsets and stabilize proximal synapses. The interconnected relationship between synapses and the ECM in which they are embedded suggests that alterations in one structure necessarily affect the dynamics of the other, and microglia may need to sculpt the matrix to modify the synapses within. Here, we provide an overview of the microglial regulation of synapses, perisynaptic matrix, and PNNs, propose candidate mechanisms by which these structures may be modified, and present the implications of such modifications in normal brain homeostasis and in disease.

Computational thinking (CT) is considered an essential literacy skill for all children to develop, yet conceptual, practical, and empirical work with preschool-age children is scarce. A particular gap in the research is how CT instruction should be enacted (e.g., free play, guided play, levels of scaffolding, degree of child-initiated activities, and structure of programming tasks). Therefore, we aimed to describe what preschool children's CT experiences are like when button-operated robots are introduced into their guided play. This interpretive phenomenological study applied the Mosaic Approach to explore the emergence of CT skills during guided play with a button-operated robot (Bee-Bot). Participants were 29 preschool-age children from an early childhood education center in the northeastern United States. Data sources included audio-visual recordings, observations, child focus groups, and child-generated artifacts. The findings suggest children constructed meaning across the CT dimensions, connected with others through dialogue and negotiation, and used guidance from adults to extend their learning.

When implemented appropriately, computational thinking (CT) experiences in early childhood settings build essential literacy skills and foster initial explorations of sequencing, engineering design principles, and cause-and-effect relationships. While existing research explores CT in K-12 settings, there is insufficient research documenting the true scope of CT skills for preschool-age children (ages 3-5 years old). Thus, the paucity of research in this emerging area warranted a scoping review approach. This scoping review surveys existing CT studies with preschool-age participants and maps what is known of CT learning experience design, intended educational outcomes, and CT study design. Evidence from the reviewed articles (n = 17) indicate most studies used physical kits, task-oriented activities, and varying experience timeframes and adult scaffolding. Most studies focused on learning sequencing and events with few embedding remixing and reusing skills. Additionally, studies primarily implemented pre-post research design approaches, and few utilized qualitative methods. The analysis of the reviewed articles indicates gaps exist in CT experience designs, scope of CT interventions, and CT tool research and development. We conclude with recommendations for closing the knowledge gaps by providing specific future research directions.

Turnover is particularly worrisome in Early Head Start/Head Start (EHS/HS) where consistency in relational-care systems and stable environments are critical quality indicators. Key adults in EHS/HS settings including teachers, administrators, specialists, and auxiliary personnel impact young children and families by providing care, education, and essential resources. This study addressed risk of turnover (job satisfaction, intentions to remain in position or profession) in a nation-wide sample of EHS/HS professionals (N = 1,076) through the lens of early childhood professional well-being (ECPW). Research Findings: Three ECPW factors were identified: Community Belonging (CB; [alpha] = 0.90); Safety & Security (SS; [alpha] = 0.84); and Professional Identity (PI; [alpha] = 0.76). ANOVA results indicated administrators, and professionals with college and graduate degrees, and those over 20 years of experience in their settings and the field experienced the highest levels of ECPW. SEM confirmed that ECPW was predictive of risk of turnover. Practice and Policy: Results helped refine researchers' earlier conceptualizations of ECPW. Implications of ECPW and risk of turnover in EHS/HS settings are discussed.

Background: Limited information is available for patients with breast cancer (BC) and coronavirus disease 2019 (COVID-19), especially among underrepresented racial/ethnic populations. Methods: This is a COVID-19 and Cancer Consortium (CCC19) registry-based retrospective cohort study of females with active or history of BC and laboratory-confirmed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection diagnosed between March 2020 and June 2021 in the US. Primary outcome was COVID-19 severity measured on a five-level ordinal scale, including none of the following complications, hospitalization, intensive care unit admission, mechanical ventilation, and all-cause mortality. Multivariable ordinal logistic regression model identified characteristics associated with COVID-19 severity. Results: 1383 female patient records with BC and COVID-19 were included in the analysis, the median age was 61 years, and median follow-up was 90 days. Multivariable analysis revealed higher odds of COVID-19 severity for older age (aOR per decade, 1.48 [95% CI, 1.32–1.67]); Black patients (aOR 1.74; 95 CI 1.24–2.45), Asian Americans and Pacific Islander patients (aOR 3.40; 95 CI 1.70–6.79) and Other (aOR 2.97; 95 CI 1.71–5.17) racial/ethnic groups; worse ECOG performance status (ECOG PS ≥2: aOR, 7.78 [95% CI, 4.83–12.5]); pre-existing cardiovascular (aOR, 2.26 [95% CI, 1.63–3.15])/pulmonary comorbidities (aOR, 1.65 [95% CI, 1.20–2.29]); diabetes mellitus (aOR, 2.25 [95% CI, 1.66–3.04]); and active and progressing cancer (aOR, 12.5 [95% CI, 6.89–22.6]). Hispanic ethnicity, timing, and type of anti-cancer therapy modalities were not significantly associated with worse COVID-19 outcomes. The total all-cause mortality and hospitalization rate for the entire cohort was 9% and 37%, respectively however, it varied according to the BC disease status. Conclusions: Using one of the largest registries on cancer and COVID-19, we identified patient and BC-related factors associated with worse COVID-19 outcomes. After adjusting for baseline characteristics, underrepresented racial/ethnic patients experienced worse outcomes compared to non-Hispanic White patients. Funding: This study was partly supported by National Cancer Institute grant number P30 CA068485 to Tianyi Sun, Sanjay Mishra, Benjamin French, Jeremy L Warner; P30-CA046592 to Christopher R Friese; P30 CA023100 for Rana R McKay; P30-CA054174 for Pankil K Shah and Dimpy P Shah; KL2 TR002646 for Pankil Shah and the American Cancer Society and Hope Foundation for Cancer Research (MRSG-16-152-01-CCE) and P30-CA054174 for Dimpy P Shah. REDCap is developed and supported by Vanderbilt Institute for Clinical and Translational Research grant support (UL1 TR000445 from NCATS/NIH). The funding sources had no role in the writing of the manuscript or the decision to submit it for publication. Clinical trial number: CCC19 registry is registered on ClinicalTrials.gov, NCT04354701.

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Abstract Background Urinary and faecal metabolic profiling have been extensively studied in gastrointestinal diseases as potential diagnostic markers, and to enhance our understanding of the intestinal microbiome in the pathogenesis these conditions. The impact of bowel cleansing on the microbiome has been investigated in several studies, but limited to just one study on the faecal metabolome. Aim To compare the effects of bowel cleansing on the composition of the faecal microbiome, and the urine and faecal metabolome. Methods Urine and faecal samples were obtained from eleven patients undergoing colonoscopy at baseline, and then at day 3 and week 6 after colonoscopy. 16S rRNA gene sequencing was used to analyse changes in the microbiome, and metabonomic analysis was performed using proton nuclear magnetic resonance (1H NMR) spectroscopy. Results Microbiomic analysis demonstrated a reduction in alpha diversity (Shannon index) between samples taken at baseline and three days following bowel cleansing (p = 0.002), and there was no significant difference between samples at baseline and six weeks post colonoscopy. Targeted and non-targeted analysis of urinary and faecal bacterial associated metabolites showed no significant impact following bowel cleansing. Conclusions Bowel cleansing causes a temporary disturbance in bacterial alpha diversity measured in faeces, but no significant changes in the faecal and urine metabolic profiles, suggesting that overall the faecal microbiome and its associated metabolome is resistant to the effects of an induced osmotic diarrhoea.

The ca. 355 Ma Fountain Lake Group in the Cobequid Highlands of Nova Scotia, is part of the transtensional basin fill that formed during dextral strike-slip motion between Avalonia and the Meguma terranes following the Acadian Orogeny. Paleomagnetic analysis of the Fountain Lake Group offers a paleolatitude estimate for the Laurentian accretionary margin in the Early Carboniferous as well as locality-specific paleomagnetic directions, which indicate clockwise-sense block rotations during dextral strike-slip motion along the Cobequid Fault zone. Stepwise demagnetization of 142 specimens from 20 sites in three Fountain Lake Group localities across the Cobequid Highlands (Squally Point, West Moose River, and Wentworth exposures) reveals remanence consisting of an easily removed component of probable recent origin, and more persistent components carried by magnetite and hematite, which in petrographic and electron beam analysis appear to be of primary igneous and volcanic oxidation origins, respectively. Sites from all three localities carry stable characteristic remanent magnetization (ChRM) directions that assume similar moderate downward inclinations when tilt-corrected. A Block Rotation Fisher analysis inclination-only fold test demonstrated best agreement at 90% unfolding, showing that remanence acquisition pre-dates Alleghenian deformation in the Late Carboniferous and is most likely of primary 355 Ma age. Paleomagnetic results for the Squally Point, West Moose River, and Wentworth localities show relative rotations between the blocks that are variously clockwise-rotated compared with a Laurentia cratonic reference frame. Inclinations at all three localities imply a subtropics paleolatitude for the margin (at Squally Point, 27.2[degrees] [+ or -] 9.4[degrees]; N = 7 sites), directly supporting the depicted location of Laurentia and its Appalachian accretionary margin in most Devonian to Early Carboniferous reconstructions. Key words: Avalonia, tectonic rotation, Appalachian, Laurentia, paleolatitude, Meguma terrane. Le Groupe de Fountain Lake d'environ 355 Ma des terres hautes de Cobequid, en Nouvelle-Ecosse, fait partie des depots de remplissage du bassin de transtension qui s'est forme durant le coulissage dextre des terranes dAvalonie et de Meguma l 'un par rapport a l'autre apres l'orogenese acadienne. L'analyse paleomagnetique du Groupe du Fountain Lake fournit une estimation de la paleolatitude de la marge accretionnaire laurentienne au Carbonifere precoce et des directions paleomagnetiques pour differentes localites qui indiquent des rotations horaires de blocs durant le coulissage dextre le long de la zone de la faille de Cobequid. La desaimantation par etapes de 142 specimens provenant de 20 sites dans trois localites du Groupe de Fountain Lake dans les terres hautes de Cobequid (affleurements de Squally Point, West Moose River et Wentworth) revele une remanence consistant en une composante facile a enlever probablement d'origine recente et des composantes plus persistantes portees par la magnetite et l'hematite qui, selon les analyses petrographiques et a faisceau electronique, semblent respectivement etre de source ignee primaire et avoir ete formee par oxydation volcanique. Des sites dans les trois localites presentent des directions d'aimantation remanente caracteristique (MRCh) stables qui, apres correction pour l'inclinaison, indiquent des plongees moderees semblables. Un essai de plissement integrant seulement l'inclinaison jumele a l'analyse de rotation de blocs de Fisher demontre une bonne concordance pour un depliage de 90 %, ce qui montre que l 'acquisition de la remanence est anterieure a la deformation alleghanienne au Carbonifere tardif et que son age primaire est fort probablement de 355 Ma. Les resultats paleomagnetiques pour les localites de Squally Point, West Moose River et Wentworth montrent des rotations relatives entre les blocs, qui presentent une rotation horaire variable par rapport au cadre de reference du craton laurentien. Les inclinaisons dans les trois localites indiqueraient une paleolatitude subtropicale pour la marge (a Squally Point, 27,2[degrees] [+ or -] 9,4[degrees]; N = 7 sites), ce qui appuie directement l'emplacement indique de la Laurentie et de sa marge accretionnaire appalachienne dans la plupart des reconstitutions pour l'intervalle du Devonien au Carbonifere precoce. [Traduit par la Redaction] Mots-cles: Avalonie, rotation tectonique, Appalaches, Laurentie, paleolatitude, terrane de Meguma., Introduction The amalgamation of Pangea marks the assembly of the most recent, and best-known supercontinent in Earth history. The final stages of Pangea assembly took place in the late Paleozoic [...]

Background: Coronavirus disease 2019 prompted the rapid adoption of telehealth to provide physical therapy. Patients' perceptions about telehealth physical therapy are mostly unknown. This study describes perceptions of telehealth physical therapy among patients with chronic low back pain (LBP). Methods: This study surveyed participants in an ongoing multisite clinical trial of nonpharmacological LBP treatments. Participants were asked about their willingness to use telehealth for physical therapy and with other providers and completed the PROMIS-29. Results: Surveys were received from 102 participants (mean age?=?48.5 [standard deviation; SD?=?11.6]). Thirty-six (35.3%) expressed willingness to receive telehealth physical therapy, 22 were neutral (21.6%), and 44 were unwilling (43.1%). The percentage expressing willingness for telehealth physical therapy was lower than it was for family medicine (p?0.001) or mental health (p?

Annually, approximately 27% of early childhood care and education professionals turnover, or leave their positions or the field, which may negatively impact quality in two ways. First, continuity is disrupted when teachers leave, severing important relationship bonds among teachers, children, and families. Second, professionals who are considering leaving may be unhappy and distracted, which may impact their capacity to care mindfully and appropriately for young children. This quantitative study examined the relationship of Early Childhood Professional Well-being (ECPW) to "risk of turnover" (perceived job dissatisfaction and consideration of leaving one's position and/or the field) using the Early Childhood Professional Well-being Questionnaire (ECPW-Q) with a sample of 281 caregivers/teachers and administrators of child-care programs. Three factors of well-being were identified through factor analysis: Supportive Structures -- environment and climate; Collegial Relationships -- between/among adults; and Professional Beliefs & Values -- autonomy, choices, participation in decision-making. Regression results show average ECPW predictive of risk of turnover, with Path Analysis showing specific relationships among variables. Implications for further analyses are considered along with practices and policies that decrease the risk of turnover and increase well-being to support professionals' capacity to care optimally and appropriately in their work with young children and families.

Abstract To access resources in commercial laying houses hens must move between levels with agility to avoid injury. This study considered whether providing ramps during rear improved the ability of birds to transition between levels. Twelve commercial flocks (2000 birds/flock) on a multi-age site were examined between 1 and 40 weeks of age. All birds had access to elevated perching structures from 4 days of age. Six treatment flocks were also provided with ramps during rear to facilitate access to these structures. Flocks were visited three times during rear and three times at lay to record transitioning behaviour and use of the elevated structures, together with scores for keel bone and feather damage. Ramp reared flocks used the elevated structures to a greater extent at rear (P = 0.001) and at lay, when all flocks had ramps, showed less hesitancy [i.e. pacing (P = 0.002), crouching (P = 0.001) and wing-flapping (P = 0.001)] in accessing levels. Mean levels of keel bone damage were reduced in ramp reared flocks (52%) compared with control flocks (64.8%) at 40 weeks of age (P = 0.028). The early life experience of the ramp reared flocks enabled specific learning that translated and persisted in later life and resulted in overall welfare benefits.

Abstract Background Chromothripsis is an event of genomic instability leading to complex chromosomal alterations in cancer. Frequent long-range chromatin interactions between transcription factors (TFs) and targets may promote extensive translocations and copy-number alterations in proximal contact regions through inappropriate DNA stitching. Although studies have proposed models to explain the initiation of chromothripsis, few discussed how TFs influence this process for tumor progression. Methods This study focused on genomic alterations in amplification associated regions within chromosome 17. Inter−/intra-chromosomal rearrangements were analyzed using whole genome sequencing data of breast tumors in the Cancer Genome Atlas (TCGA) cohort. Common ERα binding sites were defined based on MCF-7, T47D, and MDA-MB-134 breast cancer cell lines using univariate K-means clustering methods. Nanopore sequencing technology was applied to validate frequent rearrangements detected between ATC loci on 17q23 and an ERα hub on 20q13. The efficacy of pharmacological inhibition of a potentially druggable target gene on 17q23 was evaluated using breast cancer cell lines and patient-derived circulating breast tumor cells. Results There are five adjoining regions from 17q11.1 to 17q24.1 being hotspots of chromothripsis. Inter−/intra-chromosomal rearrangements of these regions occurred more frequently in ERα-positive tumors than in ERα-negative tumors. In addition, the locations of the rearrangements were often mapped within or close to dense ERα binding sites localized on these five 17q regions or other chromosomes. This chromothriptic event was linked to concordant upregulation of 96 loci that predominantly regulate cell-cycle machineries in advanced luminal tumors. Genome-editing analysis confirmed that an ERα hub localized on 20q13 coordinately regulates a subset of these loci localized on 17q23 through long-range chromosome interactions. One of these loci, Tousled Like Kinase 2 (TLK2) known to participate in DNA damage checkpoint control, is an actionable target using phenothiazine antipsychotics (PTZs). The antiproliferative effect of PTZs was prominent in high TLK2-expressing cells, compared to low expressing cells. Conclusion This study demonstrates a new approach for identifying tumorigenic drivers from genomic regions highly susceptible to ERα-related chromothripsis. We found a group of luminal breast tumors displaying 17q-related chromothripsis for which antipsychotics can be repurposed as treatment adjuncts.

Children engage in a multitude of reciprocal relationships of care within Early Childhood Care and Education settings; they act as both care-receivers and care-givers. In order to better understand the ways in which children construct care (i.e. as they receive it and provide it to others), this study investigated how 15 three- to five-year-olds expressed, enacted, and then subsequently described and explained their experiences of care in one preschool classroom in the Midwestern United States. Data were captured using the Mosaic approach and included observations, photographs, videos, child conferences, and book-making. Analysis of the data revealed that children constructed care in five ways: to minimize the discomfort or pain of others, support relationships, promote positive emotion, enhance healthy and safe habits, and ensure the longevity and sustainability of their shared resources. Interpretations and implications of the mosaic of care generated in the site are discussed.