CryoET reveals actin filaments within platelet microtubules.

Crosstalk between the actin and microtubule cytoskeletons is important for many cellular processes. Recent studies have shown that microtubules and F-actin can assemble to form a composite structure where F-actin occupies the microtubule lumen. Whether these cytoskeletal hybrids exist in physiological settings and how they are formed is unclear. Here, we show that the short-crossover Class I actin filament previously identified inside microtubules in human HAP1 cells is cofilin-bound F-actin. Lumenal F-actin can be reconstituted in vitro, but cofilin is not essential. Moreover, actin filaments with both cofilin-bound and canonical morphologies reside within human platelet microtubules under physiological conditions. We propose that stress placed upon the microtubule network during motor-driven microtubule looping and sliding may facilitate the incorporation of actin into microtubules.

WISP-1 Regulates Cardiac Fibrosis by Promoting Cardiac Fibroblasts' Activation and Collagen Processing.

Hypertension induces cardiac fibrotic remodelling characterised by the phenotypic switching of cardiac fibroblasts (CFs) and collagen deposition. We tested the hypothesis that Wnt1-inducible signalling pathway protein-1 (WISP-1) promotes CFs' phenotypic switch, type I collagen synthesis, and in vivo fibrotic remodelling. The treatment of human CFs (HCFs, n = 16) with WISP-1 (500 ng/mL) induced a phenotypic switch (α-smooth muscle actin-positive) and type I procollagen cleavage to an intermediate form of collagen (pC-collagen) in conditioned media after 24h, facilitating collagen maturation. WISP-1-induced collagen processing was mediated by Akt phosphorylation via integrin ß1, and disintegrin and metalloproteinase with thrombospondin motifs 2 (ADAMTS-2). WISP-1 wild-type (WISP-1+/+) mice and WISP-1 knockout (WISP-1-/-) mice (n = 5-7) were subcutaneously infused with angiotensin II (AngII, 1000 ng/kg/min) for 28 days. Immunohistochemistry revealed the deletion of WISP-1 attenuated type I collagen deposition in the coronary artery perivascular area compared to WISP-1+/+ mice after a 28-day AngII infusion, and therefore, the deletion of WISP-1 attenuated AngII-induced cardiac fibrosis in vivo. Collectively, our findings demonstrated WISP-1 is a critical mediator in cardiac fibrotic remodelling, by promoting CFs' activation via the integrin ß1-Akt signalling pathway, and induced collagen processing and maturation via ADAMTS-2. Thereby, the modulation of WISP-1 levels could provide potential therapeutic targets in clinical treatment.

Lethal multiple pterygium syndrome, large cystic hygroma, and cleft palate: Rare and severe fetal presentations of RYR1- and NEB-related congenital myopathies.

Congenital myopathies are a genetically heterogeneous group of neuromuscular disorders that commonly present with congenital hypotonia and weakness but can also present broadly. The most severe presentation is neonatal with arthrogryposis and, rarely, fetal akinesia and pterygia, features also seen in lethal multiple pterygium syndrome (LMPS). We describe two fetuses with similar phenotype, including hydrops fetalis, large cystic hygromas, bilateral talipes, and fetal akinesia in the second trimester. Genetic diagnoses were made using exome sequencing. Both fetuses had a severe form of congenital myopathy. In the first fetus, we identified two novel compound heterozygous likely pathogenic variants consistent with autosomal recessive RYR1-related congenital myopathy (congenital myopathy 1B). In the second fetus, we identified two likely pathogenic variants, one of which is novel, likely in trans consistent with a diagnosis of autosomal recessive NEB-related congenital myopathy. Reaching a genetic diagnosis for these fetuses allowed the families to receive accurate genetic counseling for future pregnancies. These fetuses highlight the genetic and phenotypic heterogeneity of LMPS, and support a broad approach to genetic testing.

Supporting Nurse Professional Development Practitioners Through an Academic-Practice Pilot Initiative.

BACKGROUND: Nurse professional development practitioners (NPDPs) support licensed nurses as they transition into practice. The NPDPs themselves benefit from opportunities to grow professionally in their role as educators. METHOD: A nursing school and hospital leadership pilot initiative was conducted to support staff development for NPDPs at a Midwestern health system. Four sessions were developed by academic educators and presented to NPDPs: educational theory, backward curricular design, active learning strategies, and assessment and evaluation principles. RESULTS: The NPDPs who attended the seminar indicated that the program objectives were met and identified at least one change they planned to make in planning, course design, or evaluation. In addition, they requested future professional development opportunities. Planning and implementation of this pilot educational seminar provided valuable content for NPDPs. CONCLUSION: This pilot model can strengthen academic-practice partnerships and support ongoing staff development. [J Contin Educ Nurs. 2024;55(5):253-256.].

Physiological responses of scallops and mussels to environmental variability: Implications for future shellfish aquaculture.

Puget Sound (Washington, USA) is a large estuary, known for its profitable shellfish aquaculture industry. However, in the past decade, scientists have observed strong acidification, hypoxia, and temperature anomalies in Puget Sound. These co-occurring environmental stressors are a threat to marine ecosystems and shellfish aquaculture. Our research assesses how environmental variability in Puget Sound impacts two ecologically and economically important bivalves, the purple-hinge rock scallop (Crassodoma gigantea) and Mediterranean mussel (Mytilus galloprovincialis). Our study examines the effect of depth and seasonality on the physiology of these two important bivalves to gain insight into ideal grow-out conditions in an aquaculture setting, improving the yield and quality of this sustainable protein source. To do this, we used Hood Canal (located in Puget Sound) as a natural multiple-stressor laboratory, which allowed us to study acclimatization capacity of shellfish in their natural habitat and provide the aquaculture industry information about differences in growth rate, shell strength, and nutritional sources across depths and seasons. Bivalves were outplanted at two depths (5 and 30 m) and collected after 3.5 and 7.5 months. To maximize mussel and scallop growth potential in an aquaculture setting, our results suggest outplanting at 5 m depth, with more favorable oxygen and pH levels. Mussel shell integrity can be improved by placing out at 5 m, regardless of season, however, there were no notable differences in shell strength between depths in scallops. For both species, δ13C values were lowest at 5 m in the winter and δ15N was highest at 30 m regardless of season. Puget Sound's combination of naturally and anthropogenically acidified conditions is already proving to be a challenge for shellfish farmers. Our study provides crucial information to farmers to optimize aquaculture grow-out as we begin to navigate the impacts of climate change.

Clinical Judgment Development and Assessment in Clinical Nursing Education.

BACKGROUND: There is a gap in evidence for effective clinical teaching strategies to promote nursing students' development of clinical judgment (CJ). PURPOSE: This study examined the effect of active learning strategies (ALSs) during clinical postconference sessions on the CJ of undergraduate nursing students. METHODS: This quasi-experimental study of ALSs or standard postconferences used the Lasater Clinical Judgment Rubric (LCJR) to rate CJ at 3 time points. All faculty members and students were trained on use of the LCJR. Ratings were done by both faculty and students. RESULTS: All ratings of CJ improved over time ( P < .001). Ratings of CJ varied between the control and intervention groups. CONCLUSIONS: The findings of this study contribute to knowledge regarding the effectiveness of clinical teaching strategies for CJ and preparation for entry to nursing practice.

Mechanobiology of the endothelium in vascular health and disease: in vitro shear stress models.

In recent years, there has been growing evidence that vascular pathologies arise in sites experiencing an altered haemodynamic environment. Fluid shear stress (FSS) is an important contributor to vascular homeostasis and regulates endothelial cell (EC) gene expression, morphology, and behaviour through specialised mechanosensitive signalling pathways. The presence of an altered FSS profile is a pathological characteristic of many vascular diseases, with the most established example being the preferential localisation of atherosclerotic plaque development. However, the precise haemodynamic contributions to other vascular pathologies including coronary artery vein graft failure remains poorly defined. To evaluate potential novel therapeutics for the treatment of vascular diseases via targeting EC behaviour, it is important to undertake in vitro experiments using appropriate culture conditions, particularly FSS. There are a wide range of in vitro models used to study the effect of FSS on the cultured endothelium, each with the ability to generate FSS flow profiles through which the investigator can control haemodynamic parameters including flow magnitude and directionality. An important consideration for selection of an appropriate model of FSS exposure is the FSS profile that the model can generate, in comparison to the physiological and pathophysiological haemodynamic environment of the vessel of interest. A resource bringing together the haemodynamic environment characteristic of atherosclerosis pathology and the flow profiles generated by in vitro methods of applying FSS would be beneficial to researchers when selecting the appropriate model for their research. Consequently, here we summarise the widely used methods of exposing cultured endothelium to FSS, the flow profile they generate and their advantages and limitations in investigating the pathological contribution of altered FSS to vascular disease and evaluating novel therapeutic targets for the treatment and prevention of vascular disease.

Enhancing the Nursing Student Experience: A Mixed-Methods Approach.

BACKGROUND: There is a lack of published literature linking the nursing student experience and program evaluation relationship. A mixed-methods approach was used to improve the nursing student experience through faculty, staff, and student engagement. METHOD: A student experience initiative was launched in 2016. A 3-year plan was developed by an external consultant and steering committee. A process to prioritize improvement needs across all programs was used, with program-specific plans, faculty engagement, and results in a graduate-entry nursing program. RESULTS: Student satisfaction ratings on exit surveys increased from 1 of 10 at the start of the initiative in 2016 to 8 of 10 in 2021. Continuous improvement and engagement are needed to maintain student satisfaction in nursing education. CONCLUSION: This initiative can serve as a framework for other educational partners to consider adopting to enhance the nursing student experience and improve program outcomes. [J Nurs Educ. 2022;61(5):276-279.].

Sphingosine-1-phosphate modulates PAR1-mediated human platelet activation in a concentration-dependent biphasic manner.

Sphingosine 1-phosphate (S1P) is a bioactive signalling sphingolipid that is increased in diseases such as obesity and diabetes. S1P can modulate platelet function, however the direction of effect and S1P receptors (S1PRs) involved are controversial. Here we describe the role of S1P in regulating human platelet function and identify the receptor subtypes responsible for S1P priming. Human platelets were treated with protease-activated receptor 1 (PAR-1)-activating peptide in the presence or absence of S1P, S1PR agonists or antagonists, and sphingosine kinases inhibitors. S1P alone did not induce platelet aggregation but at low concentrations S1P enhanced PAR1-mediated platelet responses, whereas PAR1 responses were inhibited by high concentrations of S1P. This biphasic effect was mimicked by pan-S1PR agonists. Specific agonists revealed that S1PR1 receptor activation has a positive priming effect, S1PR2 and S1PR3 have no effect on platelet function, whereas S1PR4 and S1PR5 receptor activation have an inhibitory effect on PAR-1 mediated platelet function. Although platelets express both sphingosine kinase 1/2, enzymes which phosphorylate sphingosine to produce S1P, only dual and SphK2 inhibition reduced platelet function. These results support a role for SphK2-mediated S1P generation in concentration-dependent positive and negative priming of platelet function, through S1PR1 and S1PR4/5 receptors, respectively.

Characterization of Subcellular Organelles in Cortical Perisynaptic Astrocytes.

Perisynaptic astrocytic processes (PAPs) carry out several different functions, from metabolite clearing to control of neuronal excitability and synaptic plasticity. All these functions are likely orchestrated by complex cellular machinery that resides within the PAPs and relies on a fine interplay between multiple subcellular components. However, traditional transmission electron microscopy (EM) studies have found that PAPs are remarkably poor of intracellular organelles, failing to explain how such a variety of PAP functions are achieved in the absence of a proportional complex network of intracellular structures. Here, we use serial block-face scanning EM to reconstruct and describe in three dimensions PAPs and their intracellular organelles in two different mouse cortical regions. We described five distinct organelles, which included empty and full endosomes, phagosomes, mitochondria, and endoplasmic reticulum (ER) cisternae, distributed within three PAPs categories (branches, branchlets, and leaflets). The majority of PAPs belonged to the leaflets category (~60%), with branchlets representing a minority (~37%). Branches were rarely in contact with synapses (<3%). Branches had a higher density of mitochondria and ER cisternae than branchlets and leaflets. Also, branches and branchlets displayed organelles more frequently than leaflets. Endosomes and phagosomes, which accounted for more than 60% of all the organelles detected, were often associated with the same PAP. Likewise, mitochondria and ER cisternae, representing ~40% of all organelles were usually associated. No differences were noted between the organelle distribution of the somatosensory and the anterior cingulate cortex. Finally, the organelle distribution in PAPs did not largely depend on the presence of a spine apparatus or a pre-synaptic mitochondrion in the synapse that PAPs were enwrapping, with some exceptions regarding the presence of phagosomes and ER cisternae, which were slightly more represented around synapses lacking a spine apparatus and a presynaptic mitochondrion, respectively. Thus, PAPs contain several subcellular organelles that could underlie the diverse astrocytic functions carried out at central synapses.

Graduate-Entry Education for Nonnurses: Preparation, Pathways, and Progress.

AIM: The aim of the study was to review education and role preparation in US direct-entry graduate nursing programs. BACKGROUND: In the last 15 years, there has been an increase in nursing programs targeting college graduates with nonnurse degrees. Educational pathways, role preparation, and doctoral study options vary. METHOD: A literature search was conducted. A secondary search of direct-entry graduate nursing programs was performed using electronic databases, including academic nursing sources and university websites. RESULTS: Forty-three journal articles were located. The secondary search identified 42 direct-entry graduate nursing programs. The majority of programs emphasize role preparation for clinical nurse leadership and advanced practice registered nursing (APRN) certification; 62 percent require doctoral preparation for entry to APRN practice. CONCLUSION: The growth of direct-entry graduate nursing programs will require educators to prepare and evaluate student outcomes to ensure competent entry to practice.

Aquaculture genomics, genetics and breeding in the United States: current status, challenges, and priorities for future research.

Advancing the production efficiency and profitability of aquaculture is dependent upon the ability to utilize a diverse array of genetic resources. The ultimate goals of aquaculture genomics, genetics and breeding research are to enhance aquaculture production efficiency, sustainability, product quality, and profitability in support of the commercial sector and for the benefit of consumers. In order to achieve these goals, it is important to understand the genomic structure and organization of aquaculture species, and their genomic and phenomic variations, as well as the genetic basis of traits and their interrelationships. In addition, it is also important to understand the mechanisms of regulation and evolutionary conservation at the levels of genome, transcriptome, proteome, epigenome, and systems biology. With genomic information and information between the genomes and phenomes, technologies for marker/causal mutation-assisted selection, genome selection, and genome editing can be developed for applications in aquaculture. A set of genomic tools and resources must be made available including reference genome sequences and their annotations (including coding and non-coding regulatory elements), genome-wide polymorphic markers, efficient genotyping platforms, high-density and high-resolution linkage maps, and transcriptome resources including non-coding transcripts. Genomic and genetic control of important performance and production traits, such as disease resistance, feed conversion efficiency, growth rate, processing yield, behaviour, reproductive characteristics, and tolerance to environmental stressors like low dissolved oxygen, high or low water temperature and salinity, must be understood. QTL need to be identified, validated across strains, lines and populations, and their mechanisms of control understood. Causal gene(s) need to be identified. Genetic and epigenetic regulation of important aquaculture traits need to be determined, and technologies for marker-assisted selection, causal gene/mutation-assisted selection, genome selection, and genome editing using CRISPR and other technologies must be developed, demonstrated with applicability, and application to aquaculture industries.Major progress has been made in aquaculture genomics for dozens of fish and shellfish species including the development of genetic linkage maps, physical maps, microarrays, single nucleotide polymorphism (SNP) arrays, transcriptome databases and various stages of genome reference sequences. This paper provides a general review of the current status, challenges and future research needs of aquaculture genomics, genetics, and breeding, with a focus on major aquaculture species in the United States: catfish, rainbow trout, Atlantic salmon, tilapia, striped bass, oysters, and shrimp. While the overall research priorities and the practical goals are similar across various aquaculture species, the current status in each species should dictate the next priority areas within the species. This paper is an output of the USDA Workshop for Aquaculture Genomics, Genetics, and Breeding held in late March 2016 in Auburn, Alabama, with participants from all parts of the United States.

From Cheerleader to Coach: The Developmental Progression of Bedside Teachers in Giving Feedback to Early Learners.

BACKGROUND: Medical students learn clinical skills at the bedside from teaching clinicians, who often learn to teach by teaching. Little is known about the process of becoming an effective clinical teacher. Understanding how teaching skills and approaches change with experience may help tailor faculty development for new teachers. Focusing on giving feedback to early learners, the authors asked: What is the developmental progression of clinician-teachers as they learn to give clinical skills feedback to medical students? METHOD: This qualitative study included longitudinal interviews with clinician-teachers over five years in a new clinical skills teaching program for preclinical medical students. Techniques derived from grounded theory were used for initial analyses. The current study focused on one theme identified in initial analyses: giving feedback to students. Transcript passages were organized by interview year, coded, and discussed in year clusters; thematic codes were compared and emergent codes developed. RESULTS: Themes related to giving feedback demonstrated a dyadic structure: characteristic of less experienced teachers versus characteristic of experienced teachers. Seven dominant dyadic themes emerged, including teacher as cheerleader versus coach, concern about student fragility versus understanding resilience, and focus on creating a safe environment versus challenging students within a safe environment. CONCLUSIONS: With consistent teaching, clinical teachers demonstrated progress in giving feedback to students in multiple areas, including understanding students' developmental trajectory and needs, developing tools and strategies, and adopting a dynamic, challenging, inclusive team approach. Ongoing teaching opportunities with targeted faculty development may help improve clinician-teachers' feedback skills and approaches.

What are the benefits of early patient contact?--A comparison of three preclinical patient contact settings.

BACKGROUND: Despite increasing attention to providing preclinical medical students with early patient experiences, little is known about associated outcomes for students. The authors compared three early patient experiences at a large American medical school where all preclinical students complete preceptorships and weekly bedside clinical-skills training and about half complete clinical, community-based summer immersion experiences. The authors asked, what are the relative outcomes and important educational components for students? METHODS: Medical students completed surveys at end of second year 2009-2011. In 2009, students compared/contrasted two of three approaches; responses framed later survey questions. In 2010 and 2011, students rated all three experiences in relevant areas (e.g., developing comfort in clinical setting). Investigators performed qualitative and quantitative analyses. RESULTS: Students rated bedside training more highly for developing comfort with clinical settings, one-on-one clinical-skills training, feedback, active clinical experience, quality of clinical training, and learning to be part of a team. They rated community clinical immersion and preceptorships more highly for understanding the life/practice of a physician and career/specialty decisions. CONCLUSIONS: Preclinical students received different benefits from the different experiences. Medical schools should define objectives of early clinical experiences and offer options accordingly. A combination of experiences may help students achieve clinical and team comfort, clinical skills, an understanding of physicians' lives/practices, and broad exposure for career decisions.

Fragility of brushite stones in shock wave lithotripsy: absence of correlation with computerized tomography visible structure.

PURPOSE: Brushite stones were imaged in vitro and then broken with shock wave lithotripsy to assess whether stone fragility correlates with internal stone structure visible on helical computerized tomography. MATERIALS AND METHODS: A total of 52 brushite calculi were scanned by micro computerized tomography, weighed, hydrated and placed in a radiological phantom. Stones were scanned using a Philips® Brilliance iCT 256 system and images were evaluated for the visibility of internal structural features. The calculi were then treated with shock wave lithotripsy in vitro. The number of shock waves needed to break each stone to completion was recorded. RESULTS: The number of shock waves needed to break each stone normalized to stone weight did not differ by HU value (p = 0.84) or by computerized tomography visible structures that could be identified consistently by all observers (p = 0.053). Stone fragility correlated highly with stone density and brushite content (each p <0.001). Calculi of almost pure brushite required the most shock waves to break. When all observations of computerized tomography visible structures were used for analysis by logistic fit, computerized tomography visible structure predicted increased stone fragility with an overall area under the ROC curve of 0.64. CONCLUSIONS: The shock wave lithotripsy fragility of brushite stones did not correlate with internal structure discernible on helical computerized tomography. However, fragility did correlate with stone density and increasing brushite mineral content, consistent with clinical experience with patients with brushite calculi. Thus, current diagnostic computerized tomography technology does not provide a means to predict when brushite stones will break well using shock wave lithotripsy.

"Can we just stop and talk?" patients value verbal communication about discharge care plans.

BACKGROUND: Studies show that hospitalized patients often do not understand their postdischarge care plan. There are few studies about patients' preferences regarding the content of discharge care plans. OBJECTIVE: To identify what patients view as essential elements of a post-hospitalization plan. DESIGN: Anonymous written survey distributed on the second day of admission to internal medicine wards. SETTING: An academic tertiary care hospital and an academic county hospital in Seattle, Washington. PATIENTS: Two hundred English-speaking adult inpatients ≥ 18 years or their proxies. RESULTS: The majority of patients (64.5%) surveyed wanted verbal discharge instructions, with only 10.5% requesting written instructions (P < 0.0001). One hundred percent of patients valued the following discharge instructions as essential: "when you need to follow-up with [primary care provider] PCP," "warning signs to call PCP," and "medicines to continue post-hospitalization." One hundred percent of patients wanted "a lot of information about my condition" and "test results," but only 39% wanted "a lot of information about my medications" (P < 0.0001). When asked to choose the most important piece of discharge instruction related to their disease, 67.5% of patients chose "lifestyle changes." One hundred percent of patients thought that personal communication between the inpatient provider and the outpatient primary care provider was "extremely important" or "essential." CONCLUSION: Patients uniformly placed high value on: 1) verbal communication about discharge care plans; 2) information about lifestyle changes for improved health; and 3) personal communication between inpatient and outpatient providers.

Stability of the infection marker struvite in urinary stone samples.

BACKGROUND AND PURPOSE: Struvite in kidney stones is an important marker for infection. In kidney stone samples, struvite is known to be prone to chemical breakdown, but no data exist on the stability of samples stored in dry form. The objective of this study was to examine stability of struvite under increasingly poor conditions of storage. MATERIALS AND METHODS: Samples of struvite kidney stones were broken to obtain 38 pieces averaging 67 mg in weight, and these were randomized into four storage conditions: Airtight containers stored in the dark, open containers in the dark, open containers in ambient light, and open containers at elevated temperature (40°C). Pieces were left for 6 months, and then analyzed for changes using micro CT and Fourier transform infrared spectroscopy (FT-IR). RESULTS: Initial samples proved to be struvite, indicating no transformation in the large specimens that had been stored in airtight containers in the dark for more than 6 years before this study. Pieces of struvite taken from these large specimens appeared unchanged by micro CT and FT-IR after being stored in closed containers for 6 months, but 8 of 9 pieces in open containers showed the presence of newberyite in surface layers, as did 10 of 10 pieces in open containers out in ambient light. All pieces stored at 40°C showed transformation of struvite, with 60% of the pieces showing the presence of amorphous phosphates, indicating complete breakdown of struvite in the surface layers of the pieces. CONCLUSION: We conclude that struvite in dry kidney stone samples is stable when the specimens are stored in airtight containers at room temperature, even after several years.