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Abstract Low Earth Orbit (LEO) has emerged as a unique environment for evaluating altered stem cell properties in microgravity. LEO has become increasingly accessible for research and development due to progress in private spaceflight. Axiom Mission 2 (Ax-2) was launched as the second all-private astronaut mission to the International Space Station (ISS). Frozen human induced pluripotent stem cells (hiPSCs) expressing green fluorescent protein (GFP) under the SOX2 promoter, as well as fibroblasts differentiated from SOX2-GFP hiPSCs, were sent to the ISS. Astronauts then thawed and seeded both cell types into commercially available 96-well plates, which provided surface tension that reduced fluid movement out of individual wells and showed that hiPSCs or hiPSC-derived fibroblasts could survive either in suspension or attached to a Matrigel substrate. Furthermore, both cell types could be transfected with red fluorescent protein (RFP)-expressing plasmid. We demonstrate that hiPSCs and hiPSC-fibroblasts can be thawed in microgravity in off-the-shelf, commercially-available cell culture hardware, can associate into 3D spheroids or grow adherently in Matrigel, and can be transfected with DNA. This lays the groundwork for future biomanufacturing experiments in space.

The paper explores how preservice ELA teachers' expectations of teaching compare to the reality of their experiences during the first year of teaching. The authors consider common concerns of beginning ELA teachers and their implications for teacher self-efficacy. The paper is informed by narrative research, which provides practical and specific insights into the lived experiences of participants. The first data set consists of reflective writings on a self-identified topic connected to ELA teaching and/or learning throughout the semester. Two, one-hour individual interviews conducted during the first year of teaching form the second data set. The paper provides empirical insights about how preservice experiences inform ELA teachers' expectations of first-year teaching and their development of self-efficacy. Their two major concerns -- classroom management and building rapport -- identified their fears and insecurities about managing disruptive students and establishing connections with students. These struggles offer a connection between expectations, experiences and self-efficacy. Likewise, they point to the need for teacher education to address preservice teachers' self-efficacy as a way to support their successful entry into the classroom. The paper includes implications for the development of increased opportunities to study and experience critical concerns of the profession. Such learning experiences offer preservice teachers meaningful opportunities to engage with experiential learning, applied practice and critical reflection before their first year in the field. The paper fulfills an identified need to study how differences between expectation and reality can be difficult for beginning ELA teachers to reconcile, a disconnect that lends itself to considerations of teachers' self-efficacy.

Abstract Unlike adult mammals, zebrafish regenerate spinal cord tissue and recover locomotor ability after a paralyzing injury. Here, we find that ependymal cells in zebrafish spinal cords produce the neurogenic factor Hb-egfa upon transection injury. Animals with hb-egfa mutations display defective swim capacity, axon crossing, and tissue bridging after spinal cord transection, associated with disrupted indicators of neuron production. Local recombinant human HB-EGF delivery alters ependymal cell cycling and tissue bridging, enhancing functional regeneration. Epigenetic profiling reveals a tissue regeneration enhancer element (TREE) linked to hb-egfa that directs gene expression in spinal cord injuries. Systemically delivered recombinant AAVs containing this zebrafish TREE target gene expression to crush injuries of neonatal, but not adult, murine spinal cords. Moreover, enhancer-based HB-EGF delivery by AAV administration improves axon densities after crush injury in neonatal cords. Our results identify Hb-egf as a neurogenic factor necessary for innate spinal cord regeneration and suggest strategies to improve spinal cord repair in mammals.

We aimed to determine whether results of our prior randomized control trial [RCT; NCT02301195, (1)] of Therapeutic Horseback Riding (THR) for children and adolescents with autism spectrum disorder (ASD) could be replicated at a different riding center and if treatment effects also included differences in the expression of associations between problem behavior and the activity of the hypothalamic-pituitary-adrenal (HPA) axis. Participants with ASD (N = 16) ages 6-16 years were randomized by nonverbal intelligence quotient to either a 10-week THR group (n = 8) or no horse interaction barn activity (BA) control group (n = 8). Outcome measures were a standard speech-language sample and caregiver-report of aberrant and social behaviors. Participants' saliva was sampled weekly at a consistent afternoon time immediately pre- and 20 min' post-condition (later assayed for cortisol). Intent-to-treat analysis revealed that compared to controls, THR participants had significant improvements in hyperactivity, and social awareness, and significant improvements at the 0.1 significance level in irritability and social communication behaviors. There were no significant improvements in number of words or new words spoken during the standard language sample. Linear mixed effects model analysis indicated that greater weekly pre-lesson irritability levels were associated with smaller post-lesson reduction in salivary cortisol levels, and greater weekly pre-lesson hyperactivity levels were associated with smaller cortisol reduction in the THR group, but not in the BA control group. The findings represent a partial replication of prior results (1), extend prior observations to include THR effects on biobehavioral relationships and suggest that cortisol could be a target mediator for THR effects on irritability and hyperactivity behaviors in youth with ASD. Clinical Trial Registration: Trial of Therapeutic Horseback Riding in Children and Adolescents with Autism Spectrum Disorder; http://clinicaltrials.gov, identifier: NCT02301195.

The immune response to SARS-CoV-2 infection is variable but has been linked to prognosis and the development of severe immunopathology. Here the authors assess a range of immune parameters in both peripheral blood and respiratory samples, providing a comparative assessment of the immune response between these compartments and their potential impact on immune-pathogenesis.

Crystallography is a major technique for determining large RNA structures. Obtaining diffraction-quality crystals has been the bottleneck. Although several RNA crystallization methods have been developed, the field strongly needs additional approaches. Here we invented an in crystallo selection strategy for identifying mutations that enhance a target RNA's crystallizability. The strategy includes constructing an RNA pool containing random mutations, obtaining crystals, and amplifying the sequences enriched by crystallization. We demonstrated a proof-of-principle application to the P4-P6 domain from the Tetrahymena ribozyme. We further determined the structures of four selected mutants. All four establish new crystal lattice contacts while maintaining the native structure. Three mutants achieve this by relocating bulges and one by making a helix more flexible. In crystallo selection provides opportunities to improve crystals of RNAs or RNA-ligand complexes. Our results also suggest that mutants may be rationally designed for crystallization by "walking" a bulge along the RNA chain.

We aimed to determine whether results of our prior randomized control trial [RCT; NCT02301195, (1)] of Therapeutic Horseback Riding (THR) for children and adolescents with autism spectrum disorder (ASD) could be replicated at a different riding center and if treatment effects also included differences in the expression of associations between problem behavior and the activity of the hypothalamic-pituitary-adrenal (HPA) axis. Participants with ASD (N = 16) ages 6-16 years were randomized by nonverbal intelligence quotient to either a 10-week THR group (n = 8) or no horse interaction barn activity (BA) control group (n = 8). Outcome measures were a standard speech-language sample and caregiver-report of aberrant and social behaviors. Participants' saliva was sampled weekly at a consistent afternoon time immediately pre- and 20 min' post-condition (later assayed for cortisol). Intent-to-treat analysis revealed that compared to controls, THR participants had significant improvements in hyperactivity, and social awareness, and significant improvements at the 0.1 significance level in irritability and social communication behaviors. There were no significant improvements in number of words or new words spoken during the standard language sample. Linear mixed effects model analysis indicated that greater weekly pre-lesson irritability levels were associated with smaller post-lesson reduction in salivary cortisol levels, and greater weekly pre-lesson hyperactivity levels were associated with smaller cortisol reduction in the THR group, but not in the BA control group. The findings represent a partial replication of prior results (1), extend prior observations to include THR effects on biobehavioral relationships and suggest that cortisol could be a target mediator for THR effects on irritability and hyperactivity behaviors in youth with ASD. Clinical Trial Registration: Trial of Therapeutic Horseback Riding in Children and Adolescents with Autism Spectrum Disorder; http://clinicaltrials.gov, identifier: NCT02301195.

ObjectivesThe common data elements (CDE) project was developed by the National Institute of Neurological Disorders and Stroke (NINDS) to provide clinical researchers with tools to improve data quality and allow for harmonization of data collected in different research studies. CDEs have been created for several neurological diseases; the aim of this project was to develop CDEs specifically curated for mitochondrial disease (Mito) to enhance clinical research.MethodsNine working groups (WGs), composed of international mitochondrial disease experts, provided recommendations for Mito clinical research. They initially reviewed existing NINDS CDEs and instruments, and developed new data elements or instruments when needed. Recommendations were organized, internally reviewed by the Mito WGs, and posted online for external public comment for a period of eight weeks. The final version was again reviewed by all WGs and the NINDS CDE team prior to posting for public use.ResultsThe NINDS Mito CDEs and supporting documents are publicly available on the NINDS CDE website ( https://commondataelements.ninds.nih.gov/ ), organized into domain categories such as Participant/Subject Characteristics, Assessments, and Examinations.ConclusionWe developed a comprehensive set of CDE recommendations, data definitions, case report forms (CRFs), and guidelines for use in Mito clinical research. The widespread use of CDEs is intended to enhance Mito clinical research endeavors, including natural history studies, clinical trial design, and data sharing. Ongoing international collaboration will facilitate regular review, updates and online publication of Mito CDEs, and support improved consistency of data collection and reporting.

Antibody responses to SARS-CoV-2 are critical in the immune response to infection, but the potential cross-reactivity to other human corona viruses is poorly appreciated. Here the authors apply a systems based approach to characterise the antibody response in pre-pandemic cohorts and assess heterotypic reactivity to SARS-CoV-2.

Study abroad programs offer undergraduate students multiple benefits by providing academic, professional and personal learning experiences that cannot be fully replicated in the domestic university setting. Recognizing the unique positioning of preservice English teachers participating in a London summer study abroad program, the author created a qualitative study to explore the implications of the preservice teachers' experiences. This article begins by connecting the potential benefits of study abroad, generally, to preservice teachers, specifically. A description of the summer study abroad program in London is followed by an explanation of the qualitative study used to examine preservice English teachers' experiences in the program. Three themes are identified and discussed -- personal independence, engagement in learning and pedagogical connection -- as well as the implications of the independence, engagement and connection developed during study abroad when the preservice teachers enter the classroom as secondary English teachers.

Children with SARS-CoV-2 infection are more likely to have mild symptoms and may be asymptomatic, but underlying reasons remain unclear. Here, the authors show cellular, cytokine and antibody response to SARS-CoV-2 infection in three children who repeatedly tested negative for the virus by PCR, despite high exposure in the household.

Studying land use change in protected areas (PAs) located in tropical forests is a major conservation priority due to high conservation value (e.g., species richness and carbon storage) here, coupled with generally high deforestation rates. Land use change researchers use a variety of land cover products to track deforestation trends, including maps they produce themselves and readily available products, such as the Global Forest Change (GFC) dataset. However, all land cover maps should be critically assessed for limitations and biases to accurately communicate and interpret results. In this study, we assess deforestation in PA complexes located in agricultural frontiers in the Amazon Basin. We studied three specific sites: Amboró and Carrasco National Parks in Bolivia, Jamanxim National Forest in Brazil, and Tambopata National Reserve and Bahuaja-Sonene National Park in Peru. Within and in 20km buffer areas around each complex, we generated land cover maps using composites of Landsat imagery and supervised classification, and compared deforestation trends to data from the GFC dataset. We then performed a dissimilarity analysis to explore the discrepancies between the two remote sensing products. Both the GFC and our supervised classification showed that deforestation rates were higher in the 20km buffer than inside the PAs and that Jamanxim National Forest had the highest deforestation rate of the PAs we studied. However, GFC maps showed consistently higher rates of deforestation than our maps. Through a dissimilarity analysis, we found that many of the inconsistencies between these datasets arise from different treatment of mixed pixels or different parameters in map creation (for example, GFC does not detect reforestation after 2012). We found that our maps underestimated deforestation while GFC overestimated deforestation, and that true deforestation rates likely fall between our two estimates. We encourage users to consider limitations and biases when using or interpreting our maps, which we make publicly available, and GFC's maps.

Conceptions of literacy in the secondary English language arts classroom today have expanded in multiple directions, moving far beyond former emphases on reading comprehension and writing ability. This article presents the efforts of two secondary English language arts teachers in the Midwestern United States to expand the meaning of literacy in their own classrooms. Using a case study approach, the authors examine each teacher's understanding of literacy, views on enacting literacy in the classroom and efforts to engage students in multiliteracies. The authors then discuss the implications of the teachers' literacy work, focusing on the teachers' individual classrooms and the preparation of secondary English teachers.

The increasing importance of technology in today's world challenges teacher educators to create technology-proficient teachers, practitioners who can utilize existing technology, learn to work with emerging technology and adapt as needed when confronted with technological issues. Acknowledging the need for technologically proficient teachers, teacher educators across the nation now infuse some degree of technological competency into the preparation of their preservice teachers. Teacher educators face several obstacles to the implementation of technology into teacher preparation, however, among them the attitude held by preservice teachers toward the technology itself. While providing opportunities for preservice teachers to engage with technology personally, academically and pedagogically, teacher educators must also consider the attitudes toward technology that influence preservice teachers' experiences with that same technology. Technology has a place not only in preservice teachers' emerging classroom pedagogy but in their professional development as well. Reflective practice is one area currently drawing on different technologies, as teacher educators incorporate a range of technology into preservice teacher reflection. Drawing from a qualitative study in which weblogs were used for voluntary preservice teacher reflective practice, this article examines preservice teacher attitudes toward a specific technology and the influence of those attitudes on the use of that technology for voluntary reflection. The researcher uses the term "voluntary reflection" to express reflection that is undertaken by choice, outside course or programmatic requirements, with all aspects of the reflective activity (such as topic, quantity, formality and medium) determined by the preservice teacher. This article opens with an overview of reflection in teacher education and the use of asynchronous communication technologies in reflective practice, with a focus on the specific technology of weblogs. The study, in which preservice teachers volunteered to use a weblog for their personal reflection while completing a teacher preparation program, is then briefly explained. Next, the preservice teachers' use of reflective weblogs is presented, followed by a discussion of the role played by individual attitudes toward technology in the use of reflective weblogs. The article concludes by considering the implications of preservice teacher attitudes toward technology for teacher educators. (Contains 1 table.)

This research examined the effects of case-based instructional strategies on the development of Pedagogical Technology Integration Content Knowledge (PTICK) in alternative teacher preparation students. The study was part of the Crossroads Project funded by the Preparing Tomorrow's Teachers for Using Technology (PT3) grant from the United States Department of Education. Thirty-three students completed a six-week course in technology integration in teaching methods at a large Southeastern university. Content analysis was used to examine student data: case responses, case reflections, and course reflections. Although there were mixed responses to the case analysis process, findings indicated that as the semester progressed the preservice teachers began to display an understanding of integrated concepts of PTICK and valued learning from the group case discussions.

Canonical primary microRNA transcripts (pri-miRNAs) are characterized by a ∼30 bp hairpin flanked by single-stranded regions. These pri-miRNAs are recognized and cleaved by the Microprocessor complex consisting of the Drosha nuclease and its obligate RNA-binding partner DGCR8. It is not well understood how the Microprocessor specifically recognizes pri-miRNA substrates. Here, we show that in addition to the well-known double-stranded RNA-binding domains, DGCR8 uses a dimeric heme-binding domain to directly contact pri-miRNAs. This RNA-binding heme domain (Rhed) directs two DGCR8 dimers to bind each pri-miRNA hairpin. The two Rhed-binding sites are located at both ends of the hairpin. The Rhed and its RNA-binding surface are important for pri-miRNA processing activity. Additionally, the heme cofactor is required for formation of processing-competent DGCR8-pri-miRNA complexes. Our study reveals a unique protein-RNA interaction central to pri-miRNA recognition. We propose a unifying model in which two DGCR8 dimers clamp a pri-miRNA hairpin using their Rheds.