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Skin wounds can be caused by traumatic lacerations or incisions which disrupt the structural and functional integrity of the skin. Wound closure and primary intention treatment of the wound as soon as possible is crucial to avoid or minimize the risk of infection that can result in a compromised healing

Skin wounds can be caused by traumatic lacerations or incisions which disrupt the structural and functional integrity of the skin. Wound closure and primary intention treatment of the wound as soon as possible is crucial to avoid or minimize the risk of infection that can result in a compromised healing rate or advanced functional intricacy. The gold standard treatment for skin wound healing is suturing. Light-activated tissue sealing is an appealing alternative to sutures as it seals the wound edges minimizing the risk of infection and scarring, especially when utilized along with biodegradable polymeric biomaterials in the wound bed. Silk fibroins can be used as a biodegradable biomaterial that possesses properties supporting cell migration and proliferation in the tissue it interacts with. In addition, histamine treatment is shown to have extensive effects on cellular functions promoting wound healing. Here, the evaluation of Laser-activated Sealants (LASE) consisting of silk fibroin films induced with Indocyanine Green dye in a wound sealed with laser in the presence of Histamine receptor agonists H1R, H2R and H4R take place. The results were evaluated using Trans-epidermal Water Loss (TEWL), histological and analytical techniques where immune cell biomarkers Arginase-1, Ly6G, iNOS, Alpha-SMA, Proliferating Cell Nuclear Antigen (PCNA), and E-Cadherin were used to study the activity of specific cells such as macrophages, neutrophils, and myofibroblasts that aid in wound healing. PBS was used as a control for histamine receptor agonists. It was found that TEWL increased when treated with H1 receptor agonists while decreasing significantly in H2R and H4R-treated wounds. Arginase-1 activity improved, while it displayed an inverse relationship compared to iNOS. H4R agonist escalated Alpha-SMA cells, while others did not have any significant difference. Ly6G activity depleted in all histamine agonists significantly, while PCNA and E-Cadherin failed to show a positive or negative effect.
ContributorsPatel, Dirghau Manishbhai (Author) / Rege, Kaushal (Thesis advisor) / Massia, Stephen (Committee member) / Brafman, David (Committee member) / Arizona State University (Publisher)
Created2022
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Description
The advent of CRISPR/Cas9 revolutionized the field of genetic engineering and gave rise to the development of new gene editing tools including prime editing. Prime editing is a versatile gene editing method that mediates precise insertions and deletions and can perform all 12 types of point mutations. In turn, prime

The advent of CRISPR/Cas9 revolutionized the field of genetic engineering and gave rise to the development of new gene editing tools including prime editing. Prime editing is a versatile gene editing method that mediates precise insertions and deletions and can perform all 12 types of point mutations. In turn, prime editing represents great promise in the design of new gene therapies and disease models where editing was previously not possible using current gene editing techniques. Despite advancements in genome modification technologies, parallel enrichment strategies of edited cells remain lagging behind in development. To this end, this project aimed to enhance prime editing using transient reporter for editing enrichment (TREE) technology to develop a method for the rapid generation of clonal isogenic cell lines for disease modeling. TREE uses an engineered BFP variant that upon a C-to-T conversion will convert to GFP after target modification. Using flow cytometry, this BFP-to-GFP conversion assay enables the isolation of edited cell populations via a fluorescent reporter of editing. Prime induced nucleotide engineering using a transient reporter for editing enrichment (PINE-TREE), pairs prime editing with TREE technology to efficiently enrich for prime edited cells. This investigation revealed PINE-TREE as an efficient editing and enrichment method compared to a conventional reporter of transfection (RoT) enrichment strategy. Here, PINE-TREE exhibited a significant increase in editing efficiencies of single nucleotide conversions, small insertions, and small deletions in multiple human cell types. Additionally, PINE-TREE demonstrated improved clonal cell editing efficiency in human induced pluripotent stem cells (hiPSCs). Most notably, PINE-TREE efficiently generated clonal isogenic hiPSCs harboring a mutation in the APOE gene for in vitro modeling of Alzheimer’s Disease. Collectively, results gathered from this study exhibited PINE-TREE as a valuable new tool in genetic engineering to accelerate the generation of clonal isogenic cell lines for applications in developmental biology, disease modeling, and drug screening.
ContributorsKostes, William Warner (Author) / Brafman, David (Thesis advisor) / Jacobs, Bertram (Committee member) / Lapinaite, Audrone (Committee member) / Tian, Xiaojun (Committee member) / Wang, Xiao (Committee member) / Arizona State University (Publisher)
Created2022
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Description
Infrastructure systems are facing non-stationary challenges that stem from climate change and the increasingly complex interactions between the social, ecological, and technological systems (SETSs). It is crucial for transportation infrastructures—which enable residents to access opportunities and foster prosperity, quality of life, and social connections—to be resilient under these non-stationary challenges.

Infrastructure systems are facing non-stationary challenges that stem from climate change and the increasingly complex interactions between the social, ecological, and technological systems (SETSs). It is crucial for transportation infrastructures—which enable residents to access opportunities and foster prosperity, quality of life, and social connections—to be resilient under these non-stationary challenges. Vulnerability assessment (VA) examines the potential consequences a system is likely to experience due to exposure to perturbation or stressors and lack of the capacity to adapt. Post-fire debris flow and heat represent particularly challenging problems for infrastructure and users in the arid U.S. West. Post-fire debris flow, which is manifested with heat and drought, produces powerful runoff threatening physical transportation infrastructures. And heat waves have devastating health effects on transportation infrastructure users, including increased mortality rates. VA anticipates the potential consequences of these perturbations and enables infrastructure stakeholders to improve the system's resilience. The current transportation climate VA—which only considers a single direct climate stressor on the infrastructure—falls short of addressing the wildfire and heat challenges. This work proposes advanced transportation climate VA methods to address the complex and multiple climate stressors and the vulnerability of infrastructure users. Two specific regions were chosen to carry out the progressive transportation climate VA: 1) the California transportation networks’ vulnerability to post-fire debris flows, and 2) the transportation infrastructure user’s vulnerability to heat exposure in Phoenix.
ContributorsLi, Rui (Author) / Chester, Mikhail V. (Thesis advisor) / Middel, Ariane (Committee member) / Hondula, David M. (Committee member) / Pendyala, Ram (Committee member) / Arizona State University (Publisher)
Created2022
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Description
Ecology has been an actively studied topic recently, along with the rapid development of human microbiota-based technology. Scientists have made remarkable progress using bioinformatics tools to identify species and analyze composition. However, a thorough understanding of interspecies interactions of microbial ecosystems is still lacking, which has been a significant obstacle

Ecology has been an actively studied topic recently, along with the rapid development of human microbiota-based technology. Scientists have made remarkable progress using bioinformatics tools to identify species and analyze composition. However, a thorough understanding of interspecies interactions of microbial ecosystems is still lacking, which has been a significant obstacle in the further development of related technologies. In this work, a genetic circuit design principle with synthetic biology approaches is developed to form two-strain microbial consortia with different inter-strain interactions. The microbial systems are well-defined and inducible. Co-culture experiment results show that our microbial consortia behave consistently with previous ecological knowledge and thus serves as excellent model systems to simulate ecosystems with similar interactions. Colony patterns also emerge when co-culturing multiple species on solid media. With the engineered microbial consortia, image-processing based methods were developed to quantify the shape of co-culture colonies and distinguish microbial consortia with different interactions. Factors that affect the population ratios were identified through induction and variations in the inoculation process. Further time-lapse experiments revealed the basic rules of colony growth, composition variation, patterning, and how spatial factors impact the co-culture colony.
ContributorsChen, Xingwen (Author) / Wang, Xiao (Thesis advisor) / Kuang, Yang (Committee member) / Tian, Xiaojun (Committee member) / Brafman, David (Committee member) / Plaisier, Christopher (Committee member) / Arizona State University (Publisher)
Created2022
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Description
Annually, approximately 1.7 million people suffer a traumatic brain injury (TBI) in the United States. After initial insult, a TBI persists as a series of molecular and cellular events that lead to cognitive and motor deficits which have no treatment. In addition, the injured brain activates the regenerative niches of

Annually, approximately 1.7 million people suffer a traumatic brain injury (TBI) in the United States. After initial insult, a TBI persists as a series of molecular and cellular events that lead to cognitive and motor deficits which have no treatment. In addition, the injured brain activates the regenerative niches of the adult brain presumably to reduce damage. The subventricular zone (SVZ) niche contains neural progenitor cells (NPCs) that generate astrocytes, oligodendrocyte, and neuroblasts. Following TBI, the injury microenvironment secretes signaling molecules like stromal cell derived factor-1a (SDF-1a). SDF-1a gradients from the injury contribute to the redirection of neuroblasts from the SVZ towards the lesion which may differentiate into neurons and integrate into existing circuitry. This repair mechanism is transient and does not lead to complete recovery of damaged tissue. Further, the mechanism by which SDF-1a gradients reach SVZ cells is not fully understood. To prolong NPC recruitment to the injured brain, exogenous SDF-1a delivery strategies have been employed. Increases in cell recruitment following stroke, spinal cord injury, and TBI have been demonstrated following SDF-1a delivery. Exogenous delivery of SDF-1a is limited by its 28-minute half-life and clearance from the injury microenvironment. Biomaterials-based delivery improves stability of molecules like SDF-1a and offer control of its release. This dissertation investigates SDF-1a delivery strategies for neural regeneration in three ways: 1) elucidating the mechanisms of spatiotemporal SDF-1a signaling across the brain, 2) developing a tunable biomaterials system for SDF-1a delivery to the brain, 3) investigating SDF-1a delivery on SVZ-derived cell migration following TBI. Using in vitro, in vivo, and in silico analyses, autocrine/paracrine signaling was necessary to produce SDF-1a gradients in the brain. Native cell types engaged in autocrine/paracrine signaling. A microfluidics device generated injectable hyaluronic-based microgels that released SDF-1a peptide via enzymatic cleavage. Microgels (±SDF-1a peptide) were injected 7 days post-TBI in a mouse model and evaluated for NPC migration 7 days later using immunohistochemistry. Initial staining suggested complex presence of astrocytes, NPCs, and neuroblasts throughout the frontoparietal cortex. Advancement of chemokine delivery was demonstrated by uncovering endogenous chemokine propagation in the brain, generating new approaches to maximize chemokine-based neural regeneration.
ContributorsHickey, Kassondra (Author) / Stabenfeldt, Sarah E (Thesis advisor) / Holloway, Julianne (Committee member) / Caplan, Michael (Committee member) / Brafman, David (Committee member) / Newbern, Jason (Committee member) / Arizona State University (Publisher)
Created2021
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Description
The RNA editing enzyme adenosine deaminase acting on double stranded RNA 2 (ADAR2) converts adenosine into inosine in regions of double stranded RNA. Here, it was discovered that this critical function of ADAR2 was dysfunctional in amyotrophic lateral sclerosis (ALS) mediated by the C9orf72 hexanucleotide repeat expansion, the most common

The RNA editing enzyme adenosine deaminase acting on double stranded RNA 2 (ADAR2) converts adenosine into inosine in regions of double stranded RNA. Here, it was discovered that this critical function of ADAR2 was dysfunctional in amyotrophic lateral sclerosis (ALS) mediated by the C9orf72 hexanucleotide repeat expansion, the most common genetic abnormality associated with ALS. Typically a nuclear protein, ADAR2 was localized in cytoplasmic accumulations in postmortem tissue from C9orf72 ALS patients. The mislocalization of ADAR2 was confirmed using immunostaining in a C9orf72 mouse model and motor neurons differentiated from C9orf72 patient induced pluripotent stem cells. Notably, the cytoplasmic accumulation of ADAR2 coexisted in neurons with cytoplasmic accumulations of TAR DNA binding protein 43 (TDP-43). Interestingly, ADAR2 overexpression in mammalian cell lines induced nuclear depletion and cytoplasmic accumulation of TDP-43, reflective of the pathology observed in ALS patients. The mislocalization of TDP-43 was dependent on the catalytic activity of ADAR2 and the ability of TDP-43 to bind directly to inosine containing RNA. In addition, TDP-43 nuclear export was significantly elevated in cells with increased RNA editing. Together these results describe a novel cellular mechanism by which alterations in RNA editing drive the nuclear export of TDP-43 leading to its cytoplasmic mislocalization. Considering the contribution of cytoplasmic TDP-43 to the pathogenesis of ALS, these findings represent a novel understanding of how the formation of pathogenic cytoplasmic TDP-43 accumulations may be initiated. Further research exploring this mechanism will provide insights into opportunities for novel therapeutic interventions.
ContributorsMoore, Stephen Philip (Author) / Sattler, Rita (Thesis advisor) / Zarnescu, Daniela (Committee member) / Brafman, David (Committee member) / Van Keuren-Jensen, Kendall (Committee member) / Mangone, Marco (Committee member) / Arizona State University (Publisher)
Created2021
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Description
The GGGGCC (G4C2) hexanucleotide repeat expansion (HRE) in the C9orf72 gene is the most common genetic abnormality associated with both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two devastatingly progressive neurodegenerative diseases. The discovery of this genetic link confirmed that ALS and FTD reside along a spectrum with clinical

The GGGGCC (G4C2) hexanucleotide repeat expansion (HRE) in the C9orf72 gene is the most common genetic abnormality associated with both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), two devastatingly progressive neurodegenerative diseases. The discovery of this genetic link confirmed that ALS and FTD reside along a spectrum with clinical and pathological commonalities. Historically understood as diseases resulting in neuronal death, the role of non-neuronal cells like astrocytes is still wholly unresolved. With evidence of cortical neurodegeneration leading to cognitive impairments in C9orf72-ALS/FTD, there is a need to investigate the role of cortical astrocytes in this disease spectrum. Here, a patient-derived induced pluripotent stem cell (iPSC) cortical astrocyte model was developed to investigate consequences of C9orf72-HRE pathogenic features in this cell type. Although there were no significant C9orf72-HRE pathogenic features in cortical astrocytes, transcriptomic, proteomic and phosphoproteomic profiles elucidated global disease-related phenotypes. Specifically, aberrant expression of astrocytic-synapse proteins and secreted factors were identified. SPARCL1, a pro-synaptogenic secreted astrocyte factor was found to be selectively decreased in C9orf72-ALS/FTD iPSC-cortical astrocytes. This finding was further validated in human tissue analyses, indicating that cortical astrocytes in C9orf72-ALS/FTD exhibit a reactive transformation that is characterized by a decrease in SPARCL1 expression. Considering the evidence for substantial astrogliosis and synaptic failure leading to cognitive impairments in C9orf72-ALS/FTD, these findings represent a novel understanding of how cortical astrocytes may contribute to the cortical neurodegeneration in this disease spectrum.
ContributorsBustos, Lynette (Author) / Sattler, Rita (Thesis advisor) / Newbern, Jason (Committee member) / Zarnescu, Daniela (Committee member) / Brafman, David (Committee member) / Mehta, Shwetal (Committee member) / Arizona State University (Publisher)
Created2023
Description

En la zona metropolitana de Phoenix, el calor urbano está afectando la salud, la seguridad y la economía y se espera que estos impactos empeoren con el tiempo. Se prevé que el número de días por encima de 110˚F aumentará más del doble para el 2060. En mayo de 2017,

En la zona metropolitana de Phoenix, el calor urbano está afectando la salud, la seguridad y la economía y se espera que estos impactos empeoren con el tiempo. Se prevé que el número de días por encima de 110˚F aumentará más del doble para el 2060. En mayo de 2017, The Nature Conservancy, el Departamento de Salud Pública del condado de Maricopa, Central Arizona Conservation Alliance, la Red de Investigación en Sostenibilidad sobre la Resiliencia Urbana a Eventos Extremos, el Centro de Investigación del Clima Urbano de Arizona State University y el Center for Whole Communities lanzaron un proceso participativo de planificación de acciones contra el calor para identificar tanto estrategias de mitigación como de adaptación a fin de reducir directamente el calor y mejorar la capacidad de los residentes para lidiar con el calor. Las organizaciones comunitarias con relaciones existentes en tres vecindarios seleccionados para la planificación de acciones contra el calor se unieron más tarde al equipo del proyecto: Phoenix Revitalization Corporation, RAILMesa y Puente Movement. Más allá de construir un plan de acción comunitario contra el calor y completar proyectos de demostración, este proceso participativo fue diseñado para desarrollar conciencia, iniciativa y cohesión social en las comunidades subrepresentadas. Asimismo el proceso de planificación de acciones contra el calor fue diseñado para servir como modelo para esfuerzos futuros de resiliencia al calor y crear una visión local, contextual y culturalmente apropiada de un futuro más seguro y saludable. El método iterativo de planificación y participación utilizado por el equipo del proyecto fortaleció las relaciones dentro y entre los vecindarios, las organizaciones comunitarias, los responsables de la toma de decisiones y el equipo núcleo, y combinó la sabiduría de la narración de historias y la evidencia científica para comprender mejor los desafíos actuales y futuros que enfrentan los residentes durante eventos de calor extremo. Como resultado de tres talleres en cada comunidad, los residentes presentaron ideas que quieren ver implementadas para aumentar su comodidad y seguridad térmica durante los días de calor extremo.

Como se muestra a continuación, las ideas de los residentes se interceptaron en torno a conceptos similares, pero las soluciones específicas variaron entre los vecindarios. Por ejemplo, a todos los vecindarios les gustaría agregar sombra a sus corredores peatonales, pero variaron las preferencias para la ubicación de las mejoras para dar sombra. Algunos vecindarios priorizaron las rutas de transporte público, otros priorizaron las rutas utilizadas por los niños en su camino a la escuela y otros quieren paradas de descanso con sombra en lugares clave. Surgieron cuatro temas estratégicos generales en los tres vecindarios: promover y educar; mejorar la comodidad/capacidad de afrontamiento; mejorar la seguridad; fortalecer la capacidad. Estos temas señalan que existen serios desafíos de seguridad contra el calor en la vida diaria de los residentes y que la comunidad, los negocios y los sectores responsables de la toma de decisión deben abordar esos desafíos.

Los elementos del plan de acción contra el calor están diseñados para incorporarse a otros esfuerzos para aliviar el calor, crear ciudades resilientes al clima y brindar salud y seguridad pública. Los socios de implementación del plan de acción contra el calor provienen de la región de la zona metropolitana de Phoenix, y se brindan recomendaciones para apoyar la transformación a una ciudad más fresca.

Para ampliar la escala de este enfoque, los miembros del equipo del proyecto recomiendan a) compromiso continuo e inversiones en estos vecindarios para implementar el cambio señalado como vital por los residentes, b) repetir el proceso de planificación de acción contra el calor con líderes comunitarios en otros vecindarios, y c) trabajar con las ciudades, los planificadores urbanos y otras partes interesadas para institucionalizar este proceso, apoyando las políticas y el uso de las métricas propuestas para crear comunidades más frescas.

ContributorsMesserschmidt, Maggie (Contributor) / Guardaro, Melissa (Contributor) / White, Jessica R. (Contributor) / Berisha, Vjollca (Contributor) / Hondula, David M. (Contributor) / Feagan, Mathieu (Contributor) / Grimm, Nancy (Contributor) / Beule, Stacie (Contributor) / Perea, Masavi (Contributor) / Ramirez, Maricruz (Contributor) / Olivas, Eva (Contributor) / Bueno, Jessica (Contributor) / Crummey, David (Contributor) / Winkle, Ryan (Contributor) / Rothballer, Kristin (Contributor) / Mocine-McQueen, Julian (Contributor) / Maurer, Maria (Artist) / Coseo, Paul (Artist) / Crank, Peter J (Designer) / Broadbent, Ashley (Designer) / McCauley, Lisa (Designer) / Nature's Cooling Systems Project (Contributor) / Nature Conservancy (U.S.) (Contributor) / Phoenix Revitalization Corporation (Contributor) / Puente Movement (Contributor) / Maricopa County (Ariz.). Department of Public Health (Contributor) / Central Arizona Conservation Alliance (Contributor) / Arizona State University. Urban Climate Research Center (Contributor) / Arizona State University. Urban Resilience to Extremes Sustainability Research Network (Contributor) / Center for Whole Communities (Contributor) / RAILmesa (Contributor) / Vitalyst Health Foundation (Funder)
Created2022
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Description

According to the Centers for Disease Control and Prevention (CDC), more people die in the U.S. from heat than from all other natural disasters combined. According to the Environmental Protection Agency (EPA), more than 1,300 deaths per year in the United States are due to extreme heat. Arizona, California and

According to the Centers for Disease Control and Prevention (CDC), more people die in the U.S. from heat than from all other natural disasters combined. According to the Environmental Protection Agency (EPA), more than 1,300 deaths per year in the United States are due to extreme heat. Arizona, California and Texas are the three states with the highest burden, accounting for 43% of all heat-related deaths according to the CDC.

Although only 5% of housing in Maricopa County, Arizona, is mobile homes, approximately 30% of indoor heat-related deaths occur in these homes. Thus, the residents of mobile homes in Maricopa County are disproportionately affected by heat. Mobile home residents are extremely exposed to heat due to the high density of mobile home parks, poor construction of dwellings, lack of vegetation, socio-demographic features and not being eligible to get utility and financial assistance.

We researched numerous solutions across different domains that could help build the heat resilience of mobile home residents. As a result we found 50 different solutions for diverse stakeholders, budgets and available resources. The goal of this toolbox is to present these solutions and to explain how to apply them in order to get the most optimal result and build About this Solutions Guide People who live in mobile homes are 6 to 8 times more likely to die of heat-associated deaths. heat resilience for mobile home residents. These solutions were designed as a coordinated set of actions for everyone — individual households, mobile home residents, mobile home park owners, cities and counties, private businesses and nonprofits serving mobile home parks, and other stakeholders — to be able to contribute to heat mitigation for mobile home residents.

When we invest in a collective, coordinated suite of solutions that are designed specifically to address the heat vulnerability of mobile homes residents, we can realize a resilience dividend in maintaining affordable, feasible, liveable housing for the 20 million Americans who choose mobile homes and manufactured housing as their place to live and thrive.

ContributorsVarfalameyeva, Katsiaryna (Author) / Solís, Patricia (Author) / Phillips, Lora A. (Author) / Charley, Elisha (Author) / Hondula, David M. (Author) / Kear, Mark (Author)
Created2021
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BACKGROUND: The City of Phoenix initiated the HeatReady program in 2018 to prepare for extreme heat, as there was no official tool, framework, or mechanism at the city level to manage extreme heat. The current landscape of heat safety culture in schools, which are critical community hubs, has received less

BACKGROUND: The City of Phoenix initiated the HeatReady program in 2018 to prepare for extreme heat, as there was no official tool, framework, or mechanism at the city level to manage extreme heat. The current landscape of heat safety culture in schools, which are critical community hubs, has received less illumination. HeatReady Schools—a critical component of a HeatReady City—are those that are increasingly able to identify, prepare for, mitigate, track, and respond to the negative impacts of schoolgrounds heat. However, minimal attention has been given to formalize heat preparedness in schools to mitigate high temperatures and health concerns in schoolchildren, a heat-vulnerable population. This study set out to understand heat perceptions, (re)actions, and recommendations of key stakeholders and to identify critical themes around heat readiness. METHODS: An exploratory sequential mixed-methods case study approach was used. These methods focused on acquiring new insight on heat perceptions at elementary schools through semi-structured interviews using thematic analysis and the Delphi panel. Participants included public health professionals and school community members at two elementary schools—one public charter, one public—in South Phoenix, Arizona, a region that has been burdened historically with inequitable distribution of heat resources due to environmental racism and injustices. RESULTS: Findings demonstrated that 1) current heat safety resources are available but not fully utilized within the school sites, 2) expert opinions support that extreme heat readiness plans must account for site-specific needs, particularly education as a first step, and 3) students are negatively impacted by the effects of extreme heat, whether direct or indirect, both inside and outside the classroom. CONCLUSIONS: From key informant interviews and a Delphi panel, a list of 30 final recommendations were developed as important actions to be taken to become “HeatReady.” Future work will apply these recommendations in a HeatReady School Growth Tool that schools can tailor be to their individual needs to improve heat safety and protection measures at schools.

ContributorsShortridge, Adora (Author) / Walker, William VI (Author) / White, Dave (Committee member) / Guardaro, Melissa (Committee member) / Hondula, David M. (Committee member) / Vanos, Jennifer (Committee member) / School of Sustainability (Contributor)
Created2022-04-18