RESUMO
Inequities in COVID-19 vaccine uptake by racialized groups have been persistent throughout the vaccine rollout, leading to disparate burdens of COVID-19 outcomes. A cross-sectional study was conducted to determine COVID-19 vaccine uptake across racialized groups within the nine-county Finger Lakes region of New York State in December 2021. Cross-matching and validation were performed across multiple health information systems for the region to reduce the percentage of vaccine records with missing race information. Additionally, imputation techniques were applied to address the remaining missing values. Uptake of ≥1 dose of the COVID-19 vaccine by race was then examined. By December 2021, 828,551 individuals in our study region had received ≥1 dose of the COVID-19 vaccine, with ~25% having missing race values. Cross-matching and validation within existing records reduced this to ~7%. Uptake of ≥1 dose of a COVID-19 vaccine was greatest among individuals identifying as White, followed by those identifying as Black. The application of imputation techniques reduced the percent of missing race values to <1%; however, this reduction did not significantly change the distribution of vaccine uptake across race groups. Utilization of relevant health information systems, accompanied by imputation techniques, stands to greatly reduce the burden of missing race data within vaccine registries, facilitating accurate targeted interventions to mitigate inequities in COVID-19 vaccination.
RESUMO
HIGHLIGHTS: The DIVA scoring tool has previously been validated in the pediatric ED setting. Our project applied DIVA to an inpatient pediatric sample. DIVA may not predict likelihood of PIV success in the inpatient setting.Background: First-attempt success rate for placing pediatric peripheral intravenous (PIV) catheters ranges from to 24% to 52%. Multiple attempts can increase risk of deleterious outcomes. It is essential to screen pediatric patients appropriately to identify those who will require additional resources for successful PIV placement. Methods: A convenience sample of hospitalized pediatric patients 0 to 18 years of age on a general care unit was used in this performance improvement project. Prior to attempting PIV access, nurses completed a data collection tool that included elements of established difficult intravenous access (DIVA) tools as well as first attempt successful PIV placement. The primary outcome measure was to determine if each DIVA scoring tool is accurate in predicting the need for additional resources to achieve successful first-attempt PIV placement. The secondary outcome measure was to compare the predictive value of each DIVA scoring tool among an inpatient pediatric population. Following data exploration and cleaning, a correlation analysis was performed with logistic regression to assess DIVA score effectiveness in predicting success of PIV insertion on the first attempt. Results: Out of 133 children, 167 PIV attempts were analyzed with 150 PIV attempts included in the final data analysis. Of the 150 PIV attempts analyzed, 60% (n=90) were successful on the first attempt. Performance of prediction for first-time insertion success was comparable among all 4 DIVA scoring tools. Conclusions: None of the 4 DIVA scoring tools were superior in predicting first-time PIV placement among hospitalized children. Vein palpability was more predictive, although not statistically significant.
Assuntos
Cateterismo Periférico , Criança , Humanos , Administração Intravenosa , Pacientes Internados , Criança HospitalizadaRESUMO
Most human genetic variation is classified as variants of uncertain significance. While advances in genome editing have allowed innovation in pooled screening platforms, many screens deal with relatively simple readouts (viability, fluorescence) and cannot identify the complex cellular phenotypes that underlie most human diseases. In this paper, we present a generalizable functional genomics platform that combines high-content imaging, machine learning, and microraft isolation in a method termed "Raft-Seq". We highlight the efficacy of our platform by showing its ability to distinguish pathogenic point mutations of the mitochondrial regulator Mitofusin 2, even when the cellular phenotype is subtle. We also show that our platform achieves its efficacy using multiple cellular features, which can be configured on-the-fly. Raft-Seq enables a way to perform pooled screening on sets of mutations in biologically relevant cells, with the ability to physically capture any cell with a perturbed phenotype and expand it clonally, directly from the primary screen.
Assuntos
Edição de Genes , Genômica , Humanos , Mutação , Genômica/métodos , Fenótipo , Mitocôndrias/genéticaRESUMO
Macrophages and other immune cells are important contributors to obesity-associated inflammation; however, the cellular identities of these specific populations remain unknown. In this study, we identified individual populations of myeloid cells found in mouse epididymal/visceral adipose tissue by single-cell RNA sequencing, immunofluorescence, and flow cytometry. Multiple canonical correlation analysis identified 11 unique myeloid and myeloid-associate cell populations. In obese mice, we detected an increased percentage of monocyte-derived pro-inflammatory cells expressing Cd9 and Trem2, as well as significantly decreased percentages of multiple cell populations, including tissue-resident cells expressing Lyve1, Mafb, and Mrc1. We have identified and validated a novel myeloid/macrophage population defined by Ly6a expression, exhibiting both myeloid and mesenchymal characteristics, which increased with obesity and showed high pro-fibrotic characteristics in vitro. Our mouse adipose tissue myeloid cell atlas provides an important resource to investigate obesity-associated inflammation and fibrosis.
Assuntos
Gordura Intra-Abdominal/metabolismo , Células Mieloides/metabolismo , Obesidade/metabolismo , Análise de Sequência de RNA , Tecido Adiposo/metabolismo , Animais , Inflamação/metabolismo , Macrófagos/metabolismo , Masculino , Glicoproteínas de Membrana , Camundongos Endogâmicos C57BL , Monócitos/metabolismo , Receptores ImunológicosRESUMO
A network of intersecting molecular pathways interacts to initiate and execute axon destruction. Maximum protection against axon degeneration likely requires more than manipulation of a single target. Here, we describe the process of designing a high-throughput arrayed screening assay for the identification of key factors responsible for axon destruction and/or protection. First, we go over some existing screens in the literature, then discuss the planning, tracking, analysis, and statistics around such a screening experiment. Prioritization of perturbations may allow laboratories to cost-effectively explore the process of screening. We also present the pairing of a combinatorial drug screen with a machine learning algorithm, predicting how to best modulate neurodegenerative and neuroprotective components.
Assuntos
Axônios/fisiologia , Ensaios de Triagem em Larga Escala/métodos , Degeneração Neural/fisiopatologia , Animais , Axotomia , Sistemas CRISPR-Cas , Técnicas de Química Combinatória , Simulação por Computador , Avaliação Pré-Clínica de Medicamentos/métodos , Tentilhões/embriologia , Ensaios de Triagem em Larga Escala/instrumentação , Processamento de Imagem Assistida por Computador , Fenótipo , Controle de Qualidade , Interferência de RNA , Células Ganglionares da Retina/citologia , Sensibilidade e Especificidade , Máquina de Vetores de SuporteRESUMO
Protein domain-based approaches to analyzing sequence data are valuable tools for examining and exploring genomic architecture across genomes of different organisms. Here, we present a complete dataset of domains from the publicly available sequence data of 9,051 reference viral genomes. The data provided contain information such as sequence position and neighboring domains from 30,947 pHMM-identified domains from each reference viral genome. Domains were identified from viral whole-genome sequence using automated profile Hidden Markov Models (pHMM). This study also describes the framework for constructing "domain neighborhoods", as well as the dataset representing it. These data can be used to examine shared and differing domain architectures across viral genomes, to elucidate potential functional properties of genes, and potentially to classify viruses.
Assuntos
Bases de Dados de Proteínas , Genoma Viral , Domínios Proteicos , Cadeias de MarkovRESUMO
The blood-brain barrier (BBB) comprises the foremost protective barrier in the brain and is composed in part of a layer of microvascular endothelial cells that line the capillaries surrounding the brain. Here, we describe a human three-dimensional (3-D) cell-based model of the BBB microvascular endothelium that recapitulates properties of these cells in vivo, including physiologically relevant transcriptional profiles, the capacity to induce potent antimicrobial innate immune signaling, and the ability to resist infection by diverse RNA viruses, including members of the enterovirus (coxsackievirus B, echovirus 11, enterovirus 71, poliovirus) and flavivirus (dengue virus, Zika virus [ZIKV]) families. We show that disruption of apical tight junctions by proinflammatory cytokine tumor necrosis factor alpha (TNF-α) sensitizes 3-D-cultured BBB cells to ZIKV infection and that 3-D derived BBB cells can be used to model the transmigration of ZIKV-infected monocytes across the endothelial barrier to access underlying astrocytes. Taken together, our findings show that human BBB microvascular endothelial cells cultured in 3-D can be used to model the mechanisms by which RNA viruses access the central nervous system (CNS), which could be used for the development and screening of therapeutics to limit this event. IMPORTANCE Neurotropic viral infections are significant sources of global morbidity and mortality. The blood-brain barrier (BBB) is composed in part of a layer of microvascular endothelial cells and functions to restrict viral access to the brain. In vitro models that recapitulate many of the properties of the human BBB endothelium are lacking, particularly with respect to the unique cellular and immunological mechanisms by which these cells restrict viral infections of the brain. Here, we developed a three-dimensional cell culture model that recapitulates many of the morphological and functional properties of the BBB microvasculature and apply this model to the study of RNA virus infections. The model we describe can therefore be used to study a variety of aspects of BBB physiology, including the mechanisms by which viruses might access the CNS, and could be used for the development and screening of antiviral therapeutics to limit this important step in viral pathogenesis.
RESUMO
The integrity of long axons is essential for neural communication. Unfortunately, relatively minor stress to a neuron can cause extensive loss of this integrity. Axon degeneration is the cell-intrinsic program that actively deconstructs an axon after injury or damage. Although ultrastructural examination has revealed signs of axon degeneration in vivo, the occurrence and progression of axon degeneration in avian species have not yet been documented in vitro. Here, we use a novel cell culture system with primary embryonic zebra finch retinal ganglion cells to interrogate the properties of avian axon degeneration. First, we establish that both axotomy and a chemically induced injury (taxol and vincristine) are sufficient to initiate degeneration. These events are dependent on a late influx of calcium. In addition, as in mammals, the NAD pathway is involved, since a decrease in NMN with FK866 can reduce degeneration. Importantly, these retinal ganglion cell axons were sensitive to a pressure-induced injury, which may mimic the effect of high intraocular pressure associated with glaucoma. We have demonstrated that avian neurons undergo Wallerian degeneration in response to both physical and chemical injury. Subsequent avian studies will investigate whether blocking the degeneration pathway can protect individuals from neurodegenerative disease.
Assuntos
Axônios/fisiologia , Tentilhões/fisiologia , Degeneração Neural/fisiopatologia , Células Ganglionares da Retina/fisiologia , Estresse Fisiológico/fisiologia , Degeneração Walleriana/fisiopatologia , Análise de Variância , Animais , Axônios/patologia , Axotomia , Cálcio/metabolismo , Células Cultivadas , Modelos Animais de Doenças , Microscopia Confocal , Microscopia de Fluorescência , NAD/metabolismo , Degeneração Neural/tratamento farmacológico , Degeneração Neural/etiologia , Degeneração Neural/patologia , Paclitaxel , Pressão , Células Ganglionares da Retina/efeitos dos fármacos , Células Ganglionares da Retina/patologia , Estresse Fisiológico/efeitos dos fármacos , Vincristina , Degeneração Walleriana/tratamento farmacológico , Degeneração Walleriana/etiologia , Degeneração Walleriana/patologiaRESUMO
During mammalian pregnancy, the placenta acts as a barrier between the maternal and fetal compartments. The recently observed association between Zika virus (ZIKV) infection during human pregnancy and fetal microcephaly and other anomalies suggests that ZIKV may bypass the placenta to reach the fetus. This led us to investigate ZIKV infection of primary human trophoblasts (PHTs), which are the barrier cells of the placenta. We discovered that PHT cells from full-term placentas are refractory to ZIKV infection. In addition, medium from uninfected PHT cells protects non-placental cells from ZIKV infection. PHT cells constitutively release the type III interferon (IFN) IFNλ1, which functions in both a paracrine and autocrine manner to protect trophoblast and non-trophoblast cells from ZIKV infection. Our data suggest that for ZIKV to access the fetal compartment, it must evade restriction by trophoblast-derived IFNλ1 and other trophoblast-specific antiviral factors and/or use alternative strategies to cross the placental barrier.