Current protocols最新文献

{"title":"Visualization of the Cytoskeleton in Fixed Drosophila Embryos","authors":"Junnan Fang,&nbsp;Jovan S. Brockett,&nbsp;Weiyi Tian,&nbsp;Yuqing Hua,&nbsp;Elizabeth R. Gavis,&nbsp;Dorothy A. Lerit","doi":"10.1002/cpz1.70145","DOIUrl":"https://doi.org/10.1002/cpz1.70145","url":null,"abstract":"<p>Embryogenesis necessitates the precise orchestration of cellular events to establish tissue patterning, developmental robustness, and viability. Syncytial embryogenesis, as in <i>Drosophila melanogaster</i>, poses added challenges as the synchronous and rapid nuclear divisions prior to cellularization occur within a shared cytoplasm. While the first several rounds of nuclear divisions occur within the center of the embryo, the nuclei progressively migrate peripherally, giving rise to the syncytial blastoderm. This spatial choreography hinges upon the dynamic interplay of actin and microtubules. Actin and microtubules coordinate nuclear division and position while preventing deleterious nuclear collisions. Additionally, the cytoskeleton also facilitates the segregation of organelles and molecular cargoes, including cell fate determinants required for cellular differentiation. As development progresses, actin and microtubules drive cellularization events for both germline and somatic cell lineages. Cytoskeletal disruption causes developmental arrest and embryonic lethality, underscoring its importance for embryogenesis. Given the significance of the cytoskeleton to these events, its visualization remains a cornerstone of cell and developmental biology research. Indeed, studies of the <i>Drosophila</i> embryo cytoskeleton have yielded valuable insights into cell biological mechanisms and developmental pathways conserved in various systems. Nevertheless, achieving optimal preservation of filamentous cytoskeletal structures poses technical challenges. Here, we present an embryo fixation method tailored to enhance the visualization of actin and microtubules via standard light microscopy approaches. This protocol complements immunofluorescence and molecular labeling techniques, including the direct labeling of fluorescently tagged proteins or mRNAs. By enabling detailed analysis of the cytoskeleton, this method expands opportunities to investigate the molecular mechanisms underlying embryo development and related processes. © 2025 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Preparation of embryos for immunofluorescence of actin or microtubules</p><p><b>Basic Protocol 2</b>: Coupling immunofluorescence of the cytoskeleton with visualization of mRNAs via smFISH</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143950100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Navigating the Scholarly Literature: A Practical Guide to Searching Effectively (Without Too Much Stress)","authors":"Kathryn P. Kohn","doi":"10.1002/cpz1.70138","DOIUrl":"https://doi.org/10.1002/cpz1.70138","url":null,"abstract":"<p>Effectively searching the scholarly literature is a fundamental academic skill. However, the process can be overwhelming due to the vast amount of available research and the complexity of academic databases. This overview article provides a practical guide to navigating the literature with confidence, outlining key strategies for identifying relevant sources, refining search queries, and troubleshooting common challenges. © 2025 The Author(s). <i>Current Protocols</i> published by Wiley Periodicals LLC.</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70138","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143930204","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly Effective Chemical Ligation of DNA and l-aTNA","authors":"Hikari Okita,&nbsp;Keiji Murayama,&nbsp;Hiroyuki Asanuma","doi":"10.1002/cpz1.70140","DOIUrl":"https://doi.org/10.1002/cpz1.70140","url":null,"abstract":"<p>Chemical ligation of nucleic acids is a significant strategy for the establishment of a variety of functional biological tools and other applications. However, the conventional methodology has been suffering from low reaction efficiency or generation of unnatural structures at the ligation site. This article describes an effective chemical ligation method that connects the hydroxyl group and the monophosphate group at the nick site on the template, generating a natural phosphodiester bond. The method uses only <i>N</i>-cyanoimidazole (CNIm) and divalent metal cations to achieve chemical ligation in a template-directed manner, which can be applied to the ligations of not only DNA but also artificial nucleic acids such as <i>acyclic</i> <span>l</span>-threoninol nucleic acid (<span>l</span>-<i>a</i>TNA). Quantitative ligation is available within 10 min for <span>l</span>-<i>a</i>TNA fragments on an <span>l</span>-<i>a</i>TNA template, and within 2 hr for DNA fragments on a DNA template under optimized conditions. The effective chemical ligation system reported here will enable the development of biotechnology and nanotechnology, including exploration of <span>l</span>-<i>a</i>TNA aptamer via in vitro selection, chemical synthesis of genome-sized DNA and <span>l</span>-<i>a</i>TNA, functionalization of nanostructure, and creation of an <span>l</span>-<i>a</i>TNA-based artificial life in the future. © 2025 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Chemical ligation using CNIm and divalent metal cation</p><p><b>Basic Protocol 2</b>: Imaging analysis of ligation reaction</p><p><b>Basic Protocol 3</b>: Mass spectrometry of ligation products</p><p><b>Support Protocol</b>: Kinetic analysis</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143914533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: Streaming Long-Read Sequence Alignments for HLA Predictions Using HLAminer","authors":"René L. Warren,&nbsp;Inanc Birol","doi":"10.1002/cpz1.70144","DOIUrl":"https://doi.org/10.1002/cpz1.70144","url":null,"abstract":"<p><i>Current Protocols</i> is issuing corrections for the following protocol article.</p><p>Warren, R. L., &amp; Birol, I. (2025). Streaming long-read sequence alignments for HLA predictions using HLAminer. <i>Current Protocols</i>, <i>5</i>, e70124. doi: 10.1002/cpz1.70124</p><p>In the above-referenced article:</p><p>In Table 4, line 2, the dataset value has been changed from “NA12878, dataset A” to “NA19240, dataset A”.</p><p>The current version online now includes this correction and may be considered the authoritative version of record.</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70144","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143909025","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly Adaptable Analysis Tools for Mapping Spatial Features of Cellular Aggregates in Tissues","authors":"Andrew Sawyer,&nbsp;Nick Weingaertner,&nbsp;Ellis Patrick,&nbsp;Carl G. Feng","doi":"10.1002/cpz1.70135","DOIUrl":"https://doi.org/10.1002/cpz1.70135","url":null,"abstract":"<p>Multiplex imaging technologies have developed rapidly over the past decades. The advancement of multiplex imaging has been driven in part by the recognition that the spatial organization of cells can represent important prognostic biomarkers and that simply studying the composition of cells in diseased tissue is often insufficient. There remains a lack of tools that can perform spatial analysis at the level of cellular aggregates (a common histopathological presentation) such as tumors and granulomas, with most analysis packages focusing on smaller regions of interest and potentially missing patterns in the overall lesion structure and cellular distribution. Here, we present protocols to quantitatively describe the cellular structure of entire tissue lesions, built around two novel metrics. The Total Cell Preference Index reports whether a lesion tends to change in density in its central versus peripheral areas and can indicate the extent of necrosis across the entire lesion. The Immune Cell Preference Index then reports whether each immune cell type is located more centrally or peripherally across the entire lesion. The output of both indexes is a single number readout for simple interpretation and visualization, and these indexes can be applied to lesions of any size or shape. This simplifies cross-lesion comparison compared to traditional Euclidian distance–based analysis, which outputs multiple values for each lesion (one for output for each band used in the infiltration analysis). Additionally, this approach can be applied to any slide-scanning multiplexed imaging system, based on either protein or nucleic acid staining. Finally, the approach uses the open-source software QuPath and can be utilized by researchers with a basic understanding of QuPath, with the full analysis able to be applied to pre-generated images within 1 hr. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Image preparation and lesion selection</p><p><b>Basic Protocol 2</b>: Total Cell Preference Index and Immune Cell Preference Index</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70135","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143904887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Purification of Low-Complexity Domain Proteins FUS, EWSR1, and Their Fusions","authors":"Jesse J. Altemus,&nbsp;Michelle A. Lay,&nbsp;Valery F. Thompson,&nbsp;Jacob C. Schwartz","doi":"10.1002/cpz1.70136","DOIUrl":"https://doi.org/10.1002/cpz1.70136","url":null,"abstract":"<p>FET proteins are large multifunctional proteins that have several key roles in biology. The FET family of proteins, including FUS, EWSR1, and TAF15, play critical roles in transcription regulation, RNA processing, and DNA damage repair. These multifunctional RNA- and DNA-binding proteins are ubiquitously expressed and conserved across vertebrate species. They contain low-complexity (LC) domains that allow them to assemble and phase separate but also makes the proteins prone to aggregation. Aberrations in FET proteins, such as point mutations, aggregation, or translocations leading to fusion proteins, have been implicated in several pathologies, including frontotemporal lobar degeneration (FTLD), amyotrophic lateral sclerosis (ALS), and Ewing sarcoma. In vitro study of FET proteins is hampered by their propensity to aggregate, their disordered structure, and their susceptibility to proteolysis, making high-yield production difficult. Here, we present optimized methods for the purification of full-length FUS, EWSR1, and their fusion proteins. These protocols enable researchers to overcome issues related to aggregation and solubility, facilitating biochemical and biophysical studies of these critical yet complex proteins. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: Purification of EWSR1 and FUS proteins</p><p><b>Alternate Protocol</b>: Purification for fusion proteins</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70136","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Highly Sensitive Tissue-specific qRT-PCR-based Assay for Detection of Melanoma Cells in Tumor-Draining Lymph Nodes","authors":"Kristian M. Hargadon,&nbsp;Travis B. Goodloe III","doi":"10.1002/cpz1.70139","DOIUrl":"https://doi.org/10.1002/cpz1.70139","url":null,"abstract":"<p>Melanoma invasion of regional lymph nodes, a critical event in the progression of this disease, is well documented as a poor prognostic factor for patients with melanoma. Assessing lymph node involvement is therefore a routine part of the diagnostic workup for patients presenting with melanoma at a T stage of ≥T2a. In clinical settings, the status and degree of melanoma lymph node involvement is traditionally characterized by histopathological analysis of tissue obtained during a sentinel lymph node biopsy, a labor-intensive and costly approach that requires technically challenging sample preparation and interpretation by a trained pathologist. Alternative approaches that might reduce the financial burden and turnaround time of a sentinel lymph node biopsy workup are therefore desirable. Likewise, the ability to accurately assess lymph node invasion by melanoma in experimental settings is necessary to gain new insights into mechanisms of distant metastasis and tumor-associated immune suppression. With these applications in mind, we recently developed, and describe herein, a tissue-specific qRT-PCR-based protocol for detecting melanoma cells within tumor-draining lymph nodes. Using murine models of lymph-node-invasive and lymph-node-noninvasive melanoma cell lines and a melanin-biosynthesis-pathway-specific <i>Trp2</i> gene expression assay, we validated this method as a highly sensitive strategy for assessing lymph node involvement by melanoma. © 2025 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Growth and maintenance of murine melanoma cell cultures</p><p><b>Basic Protocol 2</b>: Extraction and quantification of RNA from murine melanoma cultures</p><p><b>Basic Protocol 3</b>: cDNA synthesis via reverse transcription</p><p><b>Basic Protocol 4</b>: Probe and primer design for TaqMan-based qPCR</p><p><b>Basic Protocol 5</b>: qPCR analysis of tissue-specific <i>Trp2</i> gene expression</p><p><b>Basic Protocol 6</b>: <i>In vitro</i> validation of qRT-PCR <i>Trp2</i> gene expression assay for detection of melanoma cells in murine whole lymph node preparations</p><p><b>Basic Protocol 7</b>: <i>In vivo</i> validation of qRT-PCR <i>Trp2</i> gene expression assay for detection of melanoma cells in murine tumor-draining lymph nodes</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143877827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryptococcus neoformans Biofilm Formation and Quantification","authors":"Oscar Romero,&nbsp;Davier Gutierrez-Gongora,&nbsp;Jennifer Geddes-McAlister","doi":"10.1002/cpz1.70133","DOIUrl":"https://doi.org/10.1002/cpz1.70133","url":null,"abstract":"<p><i>Cryptococcus neoformans</i> is an opportunistic fungal pathogen that heads the Fungal Priority Pathogen List published by the World Health Organization (WHO) in 2022. This pathogen is a primary cause of death for immunocompromised individuals (e.g., those with HIV/AIDS, the elderly, immunotherapy recipients), causing approximately 118,000 deaths yearly worldwide. <i>C. neoformans</i> relies on virulence factors that include a polysaccharide capsule, melanin, extracellular enzymes, and thermotolerance to initiate and sustain host infection. Additionally, similar to other fungal pathogens (e.g., <i>Candida albicans</i>), <i>C. neoformans</i> may develop a biofilm organization linked to more persistent cryptococcal infections. Cryptococcal biofilms are highlighted in cases of cryptococcal meningitis, in which biofilm-like structures form that are highly resistant to host immune response and to antifungal therapies. In this regard, fungal biofilm formation has become an important area of study as a means to improve our understanding of the mechanisms regulating biofilm formation and infection and to advance the discovery of antibiofilm therapeutics. To assess biofilm properties and compare across treatments, quantification and evaluation of cell viability are important. Herein, we describe a standardized method to establish a cryptococcal biofilm and quantify total biomass and cell viability. © 2025 The Author(s). <i>Current Protocols</i> published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Culturing and biofilm formation</p><p><b>Basic Protocol 2</b>: Biofilm quantification</p><p><b>Alternate Protocol</b>: Biofilm viability assay</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70133","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875570","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhancing Visual Discrimination Task: Optimized Mouse Motivation in the Touchscreen Paradigm","authors":"Jeremy Jehl,&nbsp;Fabrice Riet,&nbsp;Aline Simonet,&nbsp;Yann Herault","doi":"10.1002/cpz1.70126","DOIUrl":"https://doi.org/10.1002/cpz1.70126","url":null,"abstract":"<p>Mouse models are essential for understanding gene function, environmental interaction, and brain structure and function. This is reinforced by the ability of mice to perform complex behavioral tasks. Still, their cognitive assessments often rely on aversive paradigms, such as fear conditioning and the Morris water maze. A promising alternative is the automated touchscreen platform, which enables cognitive tests comparable to those used in humans, such as the Cambridge Neuropsychological Test Automated Battery (CANTAB). This approach enhances standardization and reduces stress by employing appetitive reinforcement. Although widely used in non-human primates, touchscreen testing remains underutilized in rodents despite its potential for cross-species cognitive research. Motivation is key to successful touchscreen tasks, often achieved through water restriction, which mice tolerate well. However, water restriction is a stressful condition, combining negative and positive reinforcement. Here, we propose an alternative that uses citric acid (CA) water to avoid classical food privation in the touchscreen paradigm to mitigate mice's stress. By creating a strong contrast with the reward, we increase the reward's positive valence. We used the touchscreen visual discrimination task to assess the effectiveness of CA water in enhancing motivation. Our results show that administering CA water on training days while allowing access to plain water on weekends reduces the learning phase duration without causing significant weight loss in wild-type C57BL/6J mice. In addition, we observed a strong commitment to performing the pattern dissociation task. This approach offers a welfare-friendly alternative for maintaining motivation in touchscreen-based cognitive tasks while minimizing stress. © 2025 Wiley Periodicals LLC.</p><p><b>Basic Protocol</b>: Pattern dissociation paradigm using sour water</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Leveraging the MethMotif Toolkit to Characterize Context-Specific Features and Roles of Methylation Sensitive Transcription Factors","authors":"Matthew Dyer,&nbsp;Gastongay Siu,&nbsp;Denis Thieffry,&nbsp;Touati Benoukraf","doi":"10.1002/cpz1.70129","DOIUrl":"https://doi.org/10.1002/cpz1.70129","url":null,"abstract":"<p>This article presents a comprehensive guide for using the MethMotif platform, which includes the MethMotif database, the TFregulomeR R package, and a new R library, Forked-TF, designed specifically for analyzing leucine-zipper transcription factors (TFs) that bind DNA as dimers. The MethMotif platform integrates transcription factor binding site (TFBS) motifs with DNA methylation profiles, providing an in-depth analysis of how methylation modulates TF binding across different cell types and conditions. The protocols are organized into three main workflows: (1) Exploration of transcription factor dimerization partners, (2) visualization of methylation-specific TF motifs using TFregulomeR, and (3) characterization of leucine-zipper TF binding patterns with a focus on dimerization. Using the platform's MethMotif database, users can retrieve ChIP-seq and DNA methylation data, intersect TFBS peak regions, and generate TFBS-methylation-informed motif logos. A case study of CEBPB in K562 cells is included to demonstrate the use of the platform, showing how to identify TF dimers, analyze their co-binding behavior, and visualize the impact of DNA methylation on binding specificity. The protocols also provide step-by-step instructions for software installation, data input formats, and interpretation of results, making it accessible to researchers with varying levels of computational expertise. Through these protocols, users can uncover how DNA methylation and TF dimerization influence gene regulatory networks, with a focus on leucine-zipper TFs in a cell-type-specific context. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Exploration of transcription factor dimerization partners</p><p><b>Support Protocol 1</b>: Software installation</p><p><b>Support Protocol 2</b>: Docker installation</p><p><b>Support Protocol 3</b>: Verifying installation</p><p><b>Basic Protocol 2</b>: Visualization of alternative cofactors</p><p><b>Basic Protocol 3</b>: Characterization of bZIP partners/cofactors</p><p><b>Basic Protocol 4</b>: Context-independent and context-dependent analysis</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70129","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143875573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}