Age-related dysregulation of the retinal transcriptome in African turquoise killifish.

Age-related vision loss caused by retinal neurodegenerative pathologies is becoming more prevalent in our ageing society. To understand the physiological and molecular impact of ageing on retinal homeostasis, we used the short-lived African turquoise killifish, a model known to naturally develop central nervous system (CNS) ageing hallmarks and vision loss. Bulk and single-cell RNA-sequencing (scRNAseq) of three age groups (6-, 12-, and 18-week-old) identified transcriptional ageing fingerprints in the killifish retina, unveiling pathways also identified in the aged brain, including oxidative stress, gliosis, and inflammageing. These findings were comparable to observations in the ageing mouse retina. Additionally, transcriptional changes in genes related to retinal diseases, such as glaucoma and age-related macular degeneration, were observed. The cellular heterogeneity in the killifish retina was characterized, confirming the presence of all typical vertebrate retinal cell types. Data integration from age-matched samples between the bulk and scRNAseq experiments revealed a loss of cellular specificity in gene expression upon ageing, suggesting potential disruption in transcriptional homeostasis. Differential expression analysis within the identified cell types highlighted the role of glial/immune cells as important stress regulators during ageing. Our work emphasizes the value of the fast-ageing killifish in elucidating molecular signatures in age-associated retinal disease and vision decline. This study contributes to the understanding of how age-related changes in molecular pathways may impact CNS health, providing insights that may inform future therapeutic strategies for age-related pathologies.

C9ORF72 deficiency results in neurodegeneration in the zebrafish retina.

Hexanucleotide repeat expansions within the gene C9ORF72 are the most common cause of the neurodegenerative diseases Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal dementia (FTD). This disease-causing expansion leads to a reduction in C9ORF72 expression levels in patients, suggesting loss of C9ORF72 function could contribute to disease. To further understand the consequences of C9ORF72 deficiency in vivo, we generated a c9orf72 mutant zebrafish line. Analysis of the adult female spinal cords revealed no appreciable neurodegenerative pathology such as loss of motor neurons, or increased levels of neuroinflammation. However, detailed examination of adult female c9orf72-/- retinas showed prominent neurodegenerative features, including a decrease in retinal thickness, gliosis, and an overall reduction in neurons of all subtypes. Analysis of rod and cone cells within the photoreceptor layer showed a disturbance in their outer segment structure and rhodopsin mis-localisation from rod outer segments to their cell bodies and synaptic terminals. Thus, C9ORF72 may play a previously unappreciated role in retinal homeostasis and suggests C9ORF72 deficiency can induce tissue specific neuronal loss.Significance statement Hexanucleotide expansions in the gene C9ORF72 are the most common cause of the Amyotrophic lateral sclerosis (ALS)/ Frontotemporal dementia (FTD) disease spectrum. The expansion reduces expression of C9ORF72 and so may play a role in neuronal loss. However, C9ORF72 loss of function has been comparatively understudied in vivo. Using the zebrafish as a model of C9ORF72 deficiency, we demonstrate that loss of C9ORF72 results in marked inflammation and neuronal loss in the aged adult zebrafish retina. Development of the retina is unaffected regardless of C9ORF72 status. This demonstrates that C9ORF72 loss of function can cause spontaneous neurodegeneration in vivo and highlights a novel role of C9ORF72 in retinal homeostasis.

Age-related dysregulation of the retinal transcriptome in African turquoise killifish.

Age-related vision loss caused by retinal neurodegenerative pathologies is becoming more prevalent in our ageing society. To understand the physiological and molecular impact of ageing on retinal homeostasis, we used the short-lived African turquoise killifish, a model known to naturally develop central nervous system (CNS) ageing hallmarks and vision loss. Bulk and single-cell RNA-sequencing (scRNA-seq) of three age groups (6-, 12-, and 18-week-old) identified transcriptional ageing fingerprints in the killifish retina, unveiling pathways also identified in the aged brain, including oxidative stress, gliosis, and inflammageing. These findings were comparable to observations in ageing mouse retina. Additionally, transcriptional changes in genes related to retinal diseases, such as glaucoma and age-related macular degeneration, were observed. The cellular heterogeneity in the killifish retina was characterised, confirming the presence of all typical vertebrate retinal cell types. Data integration from age-matched samples between the bulk and scRNA-seq experiments revealed a loss of cellular specificity in gene expression upon ageing, suggesting potential disruption in transcriptional homeostasis. Differential expression analysis within the identified cell types highlighted the role of glial/immune cells as important stress regulators during ageing. Our work emphasises the value of the fast-ageing killifish in elucidating molecular signatures in age-associated retinal disease and vision decline. This study contributes to the understanding of how age-related changes in molecular pathways may impact CNS health, providing insights that may inform future therapeutic strategies for age-related pathologies.

Nursing Professional Development Practitioners' Self-Perceived Influence.

In a quantitative study to determine nursing professional development (NPD) practitioners' self-perceived influence, influence was operationally defined as political skill. A sample of 989 NPD practitioners reported high-average overall levels of political skill. For those without formal leadership positions, networking ability, a subscale of political skill, was predicted by NPD certification and a graduate degree or higher. NPD practitioners can enhance their influence competency through education, certification, professional development, mentoring, and coaching.

Unique activities of two overlapping PAX6 retinal enhancers.

Enhancers play a critical role in development by precisely modulating spatial, temporal, and cell type-specific gene expression. Sequence variants in enhancers have been implicated in diseases; however, establishing the functional consequences of these variants is challenging because of a lack of understanding of precise cell types and developmental stages where the enhancers are normally active. PAX6 is the master regulator of eye development, with a regulatory landscape containing multiple enhancers driving the expression in the eye. Whether these enhancers perform additive, redundant or distinct functions is unknown. Here, we describe the precise cell types and regulatory activity of two PAX6 retinal enhancers, HS5 and NRE. Using a unique combination of live imaging and single-cell RNA sequencing in dual enhancer-reporter zebrafish embryos, we uncover differences in the spatiotemporal activity of these enhancers. Our results show that although overlapping, these enhancers have distinct activities in different cell types and therefore likely nonredundant functions. This work demonstrates that unique cell type-specific activities can be uncovered for apparently similar enhancers when investigated at high resolution in vivo.

Documentation of Assessments and Interventions Using a Pediatric Preoperative Risk Assessment Checklist in the Postanesthesia Care Unit: A Quality Improvement Initiative.

PURPOSE: Describe the impact of the implementation of an evidence-based pediatric preoperative risk assessment (PPRA) checklist on the frequency of postanesthesia care unit (PACU) nursing assessments and interventions in children at risk for respiratory complications during emergence from anesthesia. DESIGN: Prospective pre-/postdesign. METHODS: Pediatric perianesthesia nurses assessed 100 children preintervention according to current standard. After nurses received pediatric preoperative risk factor (PPRF) education, another 100 children were assessed postintervention using the PPRA checklist. Pre-/postpatients were unmatched for statistical purposes due to two different groups. Frequency of PACU nursing respiratory assessments/interventions was evaluated. FINDINGS: Demographic variables, risk factors, frequency of nursing assessments/interventions were summarized in pre-/postinterventions. Significant differences (P < .001) were noted between pre-/postintervention groups with increased frequency of postnursing assessments/interventions that correlated with increased risk factors and weighted risk factors. CONCLUSIONS: By identifying total PPRFs, PACU nurses used their plan of care to frequently assess and pre-emptively intervene with children who had increased risk factors to prevent or mitigate respiratory complications on emergence from anesthesia.

Effectiveness of Pediatric Competency-Based Orientation (PCBO) on Knowledge, Confidence, and Manifestation of Early Recognition of Nursing Expertise Among PACU Nurses.

PURPOSE: To evaluate the effectiveness of the American Society of Perianesthesia Nurses (ASPAN) pediatric competency-based orientation (PCBO) education program on knowledge, confidence, and early recognition of nursing expertise among perianesthesia nurses in an acute care setting. DESIGN: A quasi-experimental pre/post survey-intervention design. METHODS: Sixty perianesthesia nurses with experience ranging from less than 5 years to more than 20 years were included. A chapter review survey was completed to assess knowledge before reviewing the chapters and again after reviewing the ASPAN PCBO materials. A presurvey assessing confidence levels, decision making abilities, and early detection of knowledge regarding pediatric patients' expertise were obtained at the beginning of the study. At the end of the study, a post-study survey was completed to evaluate the effectiveness of the intervention. Random codes were assigned to each participant to ensure that the participants' information was blinded. FINDINGS: Perianesthesia nurses' knowledge increased from pre to post intervention with one of the three sets of chapters (Set 2) statistically significant. Perianesthesia nurses' confidence and recognition of nursing expertise scores increased statistically significant from pre to post intervention. Both confidence (with 33 items [P value ≤ .001]) and recognition of nursing expertise (with 16 items [P value ≤ 0.001]) were statistically significant. CONCLUSIONS: The ASPAN PCBO was shown to be statistically effective at increasing knowledge, building expertise, promoting confidence, and improving decision making skills. The plan is for the ASPAN PCBO to be incorporated into the new-hire perianesthesia orientation didactic and the competency plan.

The Impact of Preceptor Education, Experience, and Preparation on the Role.

Findings from the 2020 Association for Nursing Professional Development National Preceptor Practice Analysis study validated the Ulrich precepting model. This secondary data analysis examines the influence of preceptor training, experience, and education on perceptions of the importance of the preceptor roles, domains (knowledge and practice), and required competencies. Preceptor training compared with education and experience best predict nurses' perceived importance of precepting and its seven roles.

Findings From the 2021 Organizational Value of Nursing Professional Development Practice, Part 3: A Comparison of Pediatric and Adult Hospitals.

In 2021, the Association for Nursing Professional Development commissioned a national study to examine relationships between nursing professional development (NPD) staffing and organizational outcomes and to compare NPD staffing in pediatric and adult hospitals. This report compares data from children's and adult hospitals, which indicate that children's hospitals, in general, have significantly more staffing resources, including NPD practitioners. Insufficient data were obtained to examine relationships between NPD staffing in children's hospitals and organizational outcomes.

The Association for Nursing Professional Development National Preceptor Practice Analysis Study: An Exploration of Precepting in Perioperative Settings.

Preceptors are an important part of meeting perioperative workforce demands. A secondary analysis of data collected for the 2020 Association for Nursing Professional Development National Preceptor Practice Analysis Study focused on 400 perioperative nurse preceptors and compared their responses to nonperioperative preceptors. Most perioperative respondents had participated in preceptor training; they spent significantly more time orienting experienced nurse preceptees in the perioperative specialty and its diverse settings (eg, orthopedic surgery, open-heart surgery) than preceptors in nonperioperative settings and specialties. Perioperative preceptors spent significantly less time precepting students, suggesting the presence of an opportunity to address the nursing shortage by promoting student exposure to the perioperative setting. To align with the AORN position statements on orientation and nurse residencies, perioperative nurse leaders should ensure that properly educated preceptors are available to assist RNs who are transitioning to perioperative practice. The Ulrich Precepting Model provides an evidence-based framework for preceptor training.

A broadband thermal emission spectrum of the ultra-hot Jupiter WASP-18b.

Close-in giant exoplanets with temperatures greater than 2,000 K ('ultra-hot Jupiters') have been the subject of extensive efforts to determine their atmospheric properties using thermal emission measurements from the Hubble Space Telescope (HST) and Spitzer Space Telescope1-3. However, previous studies have yielded inconsistent results because the small sizes of the spectral features and the limited information content of the data resulted in high sensitivity to the varying assumptions made in the treatment of instrument systematics and the atmospheric retrieval analysis3-12. Here we present a dayside thermal emission spectrum of the ultra-hot Jupiter WASP-18b obtained with the NIRISS13 instrument on the JWST. The data span 0.85 to 2.85 µm in wavelength at an average resolving power of 400 and exhibit minimal systematics. The spectrum shows three water emission features (at >6σ confidence) and evidence for optical opacity, possibly attributable to H-, TiO and VO (combined significance of 3.8σ). Models that fit the data require a thermal inversion, molecular dissociation as predicted by chemical equilibrium, a solar heavy-element abundance ('metallicity', [Formula: see text] times solar) and a carbon-to-oxygen (C/O) ratio less than unity. The data also yield a dayside brightness temperature map, which shows a peak in temperature near the substellar point that decreases steeply and symmetrically with longitude towards the terminators.

Unexpected phenotypic and molecular changes of combined glucocerebrosidase and acid sphingomyelinase deficiency.

Heterozygous variants in GBA1, encoding glucocerebrosidase (GCase), are the most common genetic risk factor for Parkinson's disease (PD). Moreover, sporadic PD patients also have a substantial reduction of GCase activity. Genetic variants of SMPD1 are also overrepresented in PD cohorts, whereas a reduction of its encoded enzyme (acid sphingomyelinase or ASM) activity is linked to an earlier age of PD onset. Despite both converging on the ceramide pathway, how the combined deficiencies of both enzymes might interact to modulate PD has yet to be explored. Therefore, we created a double-knockout (DKO) zebrafish line for both gba1 (or gba) and smpd1 to test for an interaction in vivo, hypothesising an exacerbation of phenotypes in the DKO line compared to those for single mutants. Unexpectedly, DKO zebrafish maintained conventional swimming behaviour and had normalised neuronal gene expression signatures compared to those of single mutants. We further identified rescue of mitochondrial Complexes I and IV in DKO zebrafish. Despite having an unexpected rescue effect, our results confirm ASM as a modifier of GBA1 deficiency in vivo. Our study highlights the need for validating how genetic variants and enzymatic deficiencies may interact in vivo.

Early Release Science of the exoplanet WASP-39b with JWST NIRISS.

The Saturn-mass exoplanet WASP-39b has been the subject of extensive efforts to determine its atmospheric properties using transmission spectroscopy1-4. However, these efforts have been hampered by modelling degeneracies between composition and cloud properties that are caused by limited data quality5-9. Here we present the transmission spectrum of WASP-39b obtained using the Single-Object Slitless Spectroscopy (SOSS) mode of the Near Infrared Imager and Slitless Spectrograph (NIRISS) instrument on the JWST. This spectrum spans 0.6-2.8 µm in wavelength and shows several water-absorption bands, the potassium resonance doublet and signatures of clouds. The precision and broad wavelength coverage of NIRISS/SOSS allows us to break model degeneracies between cloud properties and the atmospheric composition of WASP-39b, favouring a heavy-element enhancement ('metallicity') of about 10-30 times the solar value, a sub-solar carbon-to-oxygen (C/O) ratio and a solar-to-super-solar potassium-to-oxygen (K/O) ratio. The observations are also best explained by wavelength-dependent, non-grey clouds with inhomogeneous coverageof the planet's terminator.

GliaMorph: a modular image analysis toolkit to quantify Müller glial cell morphology.

Cell morphology is crucial for all cell functions. This is particularly true for glial cells as they rely on complex shape to contact and support neurons. However, methods to quantify complex glial cell shape accurately and reproducibly are lacking. To address this, we developed the image analysis pipeline 'GliaMorph'. GliaMorph is a modular analysis toolkit developed to perform (1) image pre-processing, (2) semi-automatic region-of-interest selection, (3) apicobasal texture analysis, (4) glia segmentation, and (5) cell feature quantification. Müller glia (MG) have a stereotypic shape linked to their maturation and physiological status. Here, we characterized MG on three levels: (1) global image-level, (2) apicobasal texture, and (3) regional apicobasal vertical-to-horizontal alignment. Using GliaMorph, we quantified MG development on a global and single-cell level, showing increased feature elaboration and subcellular morphological rearrangement in the zebrafish retina. As proof of principle, we analysed expression changes in a mouse glaucoma model, identifying subcellular protein localization changes in MG. Together, these data demonstrate that GliaMorph enables an in-depth understanding of MG morphology in the developing and diseased retina.

Glia Cell Morphology Analysis Using the Fiji GliaMorph Toolkit.

Glial cells are the support cells of the nervous system. Glial cells typically have elaborate morphologies that facilitate close contacts with neighboring neurons, synapses, and the vasculature. In the retina, Müller glia (MG) are the principal glial cell type that supports neuronal function by providing a myriad of supportive functions via intricate cell morphologies and precise contacts. Thus, complex glial morphology is critical for glial function, but remains challenging to resolve at a sub-cellular level or reproducibly quantify in complex tissues. To address this issue, we developed GliaMorph as a Fiji-based macro toolkit that allows 3D glial cell morphology analysis in the developing and mature retina. As GliaMorph is implemented in a modular fashion, here we present guides to (a) setup of GliaMorph, (b) data understanding in 3D, including z-axis intensity decay and signal-to-noise ratio, (c) pre-processing data to enhance image quality, (d) performing and examining image segmentation, and (e) 3D quantification of MG features, including apicobasal texture analysis. To allow easier application, GliaMorph tools are supported with graphical user interfaces where appropriate, and example data are publicly available to facilitate adoption. Further, GliaMorph can be modified to meet users' morphological analysis needs for other glial or neuronal shapes. Finally, this article provides users with an in-depth understanding of data requirements and the workflow of GliaMorph. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Download and installation of GliaMorph components including example data Basic Protocol 2: Understanding data properties and quality 3D-essential for subsequent analysis and capturing data property issues early Basic Protocol 3: Pre-processing AiryScan microscopy data for analysis Alternate Protocol: Pre-processing confocal microscopy data for analysis Basic Protocol 4: Segmentation of glial cells Basic Protocol 5: 3D quantification of glial cell morphology.

Findings From the 2021 Organizational Value of Nursing Professional Development Practice Study, Part 1: Overview and Descriptive Data.

A 2015 study exploring the correlations between nursing professional development (NPD) staffing and organizational outcomes was replicated in 2021. Part 1 of this two-part report provides background information and descriptive data, including organizational characteristics, NPD department structures, and NPD practitioner ratios per employee, per RN, and per hospital bed. Findings reflect the proliferation of hospital systems, expansion of NPD practice to include other professions and all employees, and little change in NPD certification.

Findings From the 2021 Organizational Value of Nursing Professional Development Practice Study, Part 2: Relationships Between Nursing Professional Development Staffing and Organizational Outcomes.

A 2015 study exploring the relationship between nursing professional development (NPD) staffing and organizational outcomes was replicated in 2021. The background, including literature review, methods, and descriptive statistics, were presented in Part 1 of this report. Part 2 explores the relationships between NPD staffing and organizational outcomes. Evidence suggests that higher levels of NPD staffing are associated with many positive nursing and patient outcomes and provide financial benefit to the organization.

Meeting in the Middle: Towards Successful Multidisciplinary Bioimage Analysis Collaboration.

With an increase in subject knowledge expertise required to solve specific biological questions, experts from different fields need to collaborate to address increasingly complex issues. To successfully collaborate, everyone involved in the collaboration must take steps to "meet in the middle". We thus present a guide on truly cross-disciplinary work using bioimage analysis as a showcase, where it is required that the expertise of biologists, microscopists, data analysts, clinicians, engineers, and physicists meet. We discuss considerations and best practices from the perspective of both users and technology developers, while offering suggestions for working together productively and how this can be supported by institutes and funders. Although this guide uses bioimage analysis as an example, the guiding principles of these perspectives are widely applicable to other cross-disciplinary work.

Patients' Perceptions of Importance for Self-Administered Correct Site Surgery Checklist: A Multisite Study.

PURPOSE: The purpose of this study was to describe and validate the association between patient's self-administered correct site checklist and perceptions of importance for safe surgery. DESIGN: A multisite nonexperimental, quantitative, descriptive study. METHODS: A convenience sample of 173 adult patients from four different geographical multisite hospitals was included in the study. Inclusion criteria were age 18 to 75 years old, scheduled for surgery/procedure with laterality and ability to follow instructions. After IRB approval, investigators explained the purpose of the study, process and obtained consent from willing participants. Participants with clinical or behavioral limitations were excluded from the study. Participants completed a 24 item survey before and during surgery using a four-point Likert scale from one (not important) to four (extremely important). Descriptive data was analyzed using means, standard deviations, and percentage. All data was summarized and analyzed with STATA 12. FINDINGS: Most of the participants perceived the importance of the survey checklist items positively implying that the active engagement is an important role for safe surgery. However, a few participants reported some of the items as not important/somewhat important: "It is on my left or right side" (6.9%); "surgery on my: (state your limb) and (right or left site) (1.9-3%); "check electronic access or copy of imaging with correct name and site" (14.9%); "state your name and birthday" (4%), "check correct ID bracelet information" (2.9%) and "believe in having an active role in preventing error" (2.3%). Some participants responded, "My surgeon knows it or surgery has been scheduled".  Findings indicated that even though the importance of correct site surgery is critical for patient's surgery, a few patients reported it as noncritical and relied on healthcare team for their safety. CONCLUSIONS: This study validated the importance of the patients' perceived roles in promoting safe, correct site surgery and by engaging patients in mitigating correct site surgical errors. Therefore, inclusion of patients as an integral part of the healthcare team is necessary through education and encouragement to speak out.

Müller Glia maintain their regenerative potential despite degeneration in the aged zebrafish retina.

Ageing is a significant risk factor for degeneration of the retina. Müller glia cells (MG) are key for neuronal regeneration, so harnessing the regenerative capacity of MG in the retina offers great promise for the treatment of age-associated blinding conditions. Yet, the impact of ageing on MG regenerative capacity is unclear. Here, we show that the zebrafish retina undergoes telomerase-independent, age-related neurodegeneration but that this is insufficient to stimulate MG proliferation and regeneration. Instead, age-related neurodegeneration is accompanied by MG morphological aberrations and loss of vision. Mechanistically, yes-associated protein (Yap), part of the Hippo signalling, has been shown to be critical for the regenerative response in the damaged retina, and we show that Yap expression levels decline with ageing. Despite this, morphologically and molecularly altered aged MG retain the capacity to regenerate neurons after acute light damage, therefore, highlighting key differences in the MG response to high-intensity acute damage versus chronic neuronal loss in the zebrafish retina.