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Here, we present a cross-linking approach to covalently functionalize and stabilize DNA origami structures in a one-pot reaction. Our strategy involves adding nucleotide sequences to adjacent staple strands, so that, upon assembly of the origami structure, the extensions form short hairpin duplexes targetable by psoralen-labeled triplex-forming oligonucleotides bearing other functional groups (pso-TFOs). Subsequent irradiation with UVA light generates psoralen adducts with one or both hairpin staples leading to site-specific attachment of the pso-TFO (and attached group) to the origami with ca. 80% efficiency. Bis-adduct formation between strands in proximal hairpins further tethers the TFO to the structure and generates "superstaples" that improve the structural integrity of the functionalized complex. We show that directing cross-linking to regions outside of the origami core dramatically reduces sensitivity of the structures to thermal denaturation and disassembly by T7 RNA polymerase. We also show that the underlying duplex regions of the origami core are digested by DNase I and thus remain accessible to read-out by DNA-binding proteins. Our strategy is scalable and cost-effective, as it works with existing DNA origami structures, does not require scaffold redesign, and can be achieved with just one psoralen-modified oligonucleotide.
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Reagentes de Ligações Cruzadas , DNA , Conformação de Ácido Nucleico , Raios Ultravioleta , DNA/química , Reagentes de Ligações Cruzadas/química , Processos Fotoquímicos , Ficusina/químicaRESUMO
INTRODUCTION: The integration of whole slide imaging (WSI) and artificial intelligence (AI) with digital cytology has been growing gradually. Therefore, there is a need to evaluate the current state of digital cytology. This study aimed to determine the current landscape of digital cytology via a survey conducted as part of the American Society of Cytopathology (ASC) Digital Cytology White Paper Task Force. MATERIALS AND METHODS: A survey with 43 questions pertaining to the current practices and experiences of WSI and AI in both surgical pathology and cytology was created. The survey was sent to members of the ASC, the International Academy of Cytology (IAC), and the Papanicolaou Society of Cytopathology (PSC). Responses were recorded and analyzed. RESULTS: In total, 327 individuals participated in the survey, spanning a diverse array of practice settings, roles, and experiences around the globe. The majority of responses indicated there was routine scanning of surgical pathology slides (n = 134; 61%) with fewer respondents scanning cytology slides (n = 150; 46%). The primary challenge for surgical WSI is the need for faster scanning and cost minimization, whereas image quality is the top issue for cytology WSI. AI tools are not widely utilized, with only 16% of participants using AI for surgical pathology samples and 13% for cytology practice. CONCLUSIONS: Utilization of digital pathology is limited in cytology laboratories as compared to surgical pathology. However, as more laboratories are willing to implement digital cytology in the near future, the establishment of practical clinical guidelines is needed.
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This report highlights information and outcomes from the November 2022 ASC/IAC joint Cytology Education Symposium, an annual conference organized by the Cytology Programs Review Committee. The manuscript provides information on shared educational opportunities and practices for cytology students and other learners in anatomic pathology, discusses recruitment strategies for schools of cytology, conveys teaching resources, introduces perspectives on virtual microscopy and online learning, and transmits information about wellness of students in schools of cytology.
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Técnicas Citológicas , Instituições Acadêmicas , Simbiose , Humanos , Escolaridade , América do NorteRESUMO
Digital cytology and artificial intelligence (AI) are gaining greater adoption in the cytopathology laboratory. However, peer-reviewed real-world data and literature are lacking regarding the current clinical landscape. The American Society of Cytopathology in conjunction with the International Academy of Cytology and the Digital Pathology Association established a special task force comprising 20 members with expertise and/or interest in digital cytology. The aim of the group was to investigate the feasibility of incorporating digital cytology, specifically cytology whole slide scanning and AI applications, into the workflow of the laboratory. In turn, the impact on cytopathologists, cytologists (cytotechnologists), and cytology departments were also assessed. The task force reviewed existing literature on digital cytology, conducted a worldwide survey, and held a virtual roundtable discussion on digital cytology and AI with multiple industry corporate representatives. This white paper, presented in 2 parts, summarizes the current state of digital cytology and AI practice in global cytology practice. Part 1 of the white paper presented herein is a review and offers best practice recommendations for incorporating digital cytology into practice. Part 2 of the white paper provides a comprehensive review of AI in cytology practice along with best practice recommendations and legal considerations. Additionally, the results of a global survey regarding digital cytology are highlighted.
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Inteligência Artificial , Citodiagnóstico , Humanos , Técnicas Citológicas , Laboratórios , Fluxo de TrabalhoRESUMO
Digital cytology and artificial intelligence (AI) are gaining greater adoption in the cytology laboratory. However, peer-reviewed real-world data and literature are lacking in regard to the current clinical landscape. The American Society of Cytopathology in conjunction with the International Academy of Cytology and the Digital Pathology Association established a special task force comprising 20 members with expertise and/or interest in digital cytology. The aim of the group was to investigate the feasibility of incorporating digital cytology, specifically cytology whole slide scanning and AI applications, into the workflow of the laboratory. In turn, the impact on cytopathologists, cytologists (cytotechnologists), and cytology departments were also assessed. The task force reviewed existing literature on digital cytology, conducted a worldwide survey, and held a virtual roundtable discussion on digital cytology and AI with multiple industry corporate representatives. This white paper, presented in 2 parts, summarizes the current state of digital cytology and AI practice in global cytology practice. Part 1 of the white paper is presented as a separate paper which details a review and best practice recommendations for incorporating digital cytology into practice. Part 2 of the white paper presented here provides a comprehensive review of AI in cytology practice along with best practice recommendations and legal considerations. Additionally, the cytology global survey results highlighting current AI practices by various laboratories, as well as current attitudes, are reported.
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Inteligência Artificial , Citodiagnóstico , Humanos , Técnicas Citológicas , Laboratórios , Fluxo de TrabalhoRESUMO
This report highlights information and outcomes from the November 2022 ASC/IAC joint Cytology Education Symposium, an annual conference organized by the Cytology Programs Review Committee. The manuscript provides information on shared educational opportunities and practices for cytology students and other learners in anatomic pathology, discusses recruitment strategies for schools of cytology, conveys teaching resources, introduces perspectives on virtual microscopy and online learning, and transmits information about wellness of students in schools of cytology.
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Currículo , Simbiose , Humanos , Técnicas Citológicas , Instituições Acadêmicas , América do NorteRESUMO
INTRODUCTION: Endoscopic biopsy procedures increasingly generate multiple tissue samples from multiple sites, and frequently retrieve concurrent cytologic specimens and small core needle biopsies. There is currently lack of consensus in subspecialized practices as to whether cytopathologists or surgical pathologists should review such samples, and whether the pathology findings should be reported together or separately. MATERIALS AND METHODS: In December 2021, the American Society of Cytopathology convened the Re-Imagine Cytopathology Task Force to examine various workflows that would facilitate unified pathology reporting of concurrently obtained biopsies and improve clinical care. RESULTS AND CONCLUSIONS: This position paper summarizes the key points and highlights the advantages, challenges, and resources available to support the implementation of such workflows that result in "one procedure-one report".
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Citologia , Triagem , Humanos , Estados Unidos , Biópsia , Biópsia com Agulha de Grande Calibre , PatologistasRESUMO
Context: The shift to digital learning in medicine is well underway and in fact spurred by the COVID-19 pandemic. The didactic portion of our institution's cytotechnology (CT) education program is online and delivered to learners across the nation. With CT education elevating to the master's degree level, there is a need to expand cytologic correlation with surgical resection specimens. We also wanted to afford pathology residents the same. Methods: We developed an online cytologic-histologic correlation digital learning module (e-module) addressing thyroid fine needle aspirations (FNAs) and surgical thyroidectomy specimens which was administered as part of coursework in the CT education and pathology residency programs. The module was 35 min long and consisted of guided narration with both formative and summative interactive quizzes. After completion of the module, participants were invited to fill a brief survey comprised of multiple choice, Likert, and free response questions. This study was approved by the institutional review board. Results: The 29 respondents were comprised of 22 CT students and 7 residents. CT students had minimal experience thyroid pathology prior to the module; residents were mixed. Twenty-three (79.3%) ranked the highest tiers for learning cytopathology through this module, 24 (82.8%) for learning thyroid surgical pathology, and 25 (86.2%) for cytologic-histologic correlation. All respondents stated they would like similar activities in the future. Conclusions: Teaching cytology-histology correlation for thyroid in an electronic format was effective and well-received by participants. There is a demand for these activities among current learners, suggesting that expanding the available repertoire will be beneficial.
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Biologia Celular/educação , Certificação , Técnicas Citológicas , Educação de Pós-Graduação em Medicina , Patologistas/educação , Patologia/educação , Acreditação , Biópsia , Biologia Celular/normas , Certificação/normas , Competência Clínica , Currículo , Técnicas Citológicas/normas , Educação de Pós-Graduação em Medicina/normas , Humanos , Patologistas/normas , Patologia/normas , Especialização , Estados UnidosRESUMO
INTRODUCTION: This study reports the potential of eye-tracking technology in determining screening skills of cytotechnology (CT) students while examining digital images (DI). MATERIALS AND METHODS: Twenty-five static DI of gynecologic cytology specimens were serially displayed on a computer monitor for evaluation by 16 CT students and 3 cytotechnologists at 3 locations. During evaluation, participant's eye movements were monitored with a Mirametrix S2 eye tracker (iMotions, Boston, MA) and EyeWorks software (Eyetracking, Solana Beach, CA). Students completed the protocol at: Period1 (P1)-4 months, Period2 (P2)-7 months, Period3 (P3)-11 months during their 1-year training; and the cytotechnologists only once. A general linear mixed model was used to analyze the results. RESULTS: The proportion of agreement on interpretations for cytotechnologists, students during P1, and students during P3 were 0.83, 0.62, and 0.70 respectively. The mean task duration in seconds for cytotechnologists, students during P1, and students during P3 were 21.1, 34.6, and 24.9 respectively. The mean number of fixation points for cytotechnologists, students during P1, and students during P3 were 14.5, 52.2, and 35.3, respectively. The mean number of gaze observations of cytotechnologists, students during P1, and students during P3 on region of interest (ROI) 1 were 77.93, 181.12, and 123.83, respectively; and, ROI 2 were 38.90, 142.79, and 92.46, respectively. CONCLUSIONS: This study demonstrated that students had decreased time, number of fixation points, gaze observations on ROI, and increased agreement with the reference interpretations at the end of the training program, indicating that their screening skills were progressing towards the level of practicing cytotechnologists.
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Citodiagnóstico , Educação/métodos , Tecnologia de Rastreamento Ocular , Fixação Ocular/fisiologia , Ginecologia/educação , Pessoal de Laboratório Médico/educação , Estudantes , Competência Clínica , Escolaridade , Humanos , Estados UnidosRESUMO
INTRODUCTION: Sustaining a well-trained, competitive, and marketable cytotechnology workforce is a major goal of the American Society of Cytopathology/American Society for Clinical Pathology Workgroup Focusing on Emerging Roles in Cytopathology (ASC/ASCP Workgroup). This article describes the data collection initiatives performed by the Workgroup to confirm cytotechnologist (CT) perceived educational needs to keep pace with emerging practice changes in the workplace. In response to data collected, the Workgroup created Advanced Cytopathology Education (ACE)da unique, innovative educational resource designed to fulfill this need. MATERIALS AND METHODS: Data collection efforts included annual needs assessment surveys to gauge CT participants' preferred topics and learning modalities. These data were used to design and prioritize ACE topics. Program evaluations were also administered to evaluate participants' perceptions of program quality and effectiveness at meeting their educational needs. RESULTS: Research findings reveal CT education needs to align with emerging practice areas as reported in other Workgroup data collection efforts. The incorporation of new entry-level competencies in cytotechnology training programs prepares new CT graduates, but there is no standardized mechanism for formal, robust, and recognized ongoing education for other practicing CTs. CONCLUSIONS: This article highlights the educational endeavors carried out by the Workgroup in fulfilling the educational needs of practicing CTs as communicated through various data collection efforts. As CT responsibilities evolve, ASC and Workgroup efforts will continue to focus on providing educational support for current practicing CTs who need training in emerging practice areas.
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Técnicas Citológicas , Pessoal de Laboratório Médico , Ciência de Laboratório Médico , Humanos , Pessoal de Laboratório Médico/educação , Pessoal de Laboratório Médico/estatística & dados numéricos , Ciência de Laboratório Médico/educação , Ciência de Laboratório Médico/estatística & dados numéricosRESUMO
BACKGROUND: Nine E-learning modules (ELMs) were developed in our program using Articulate software. This study assessed our cytotechnology (CT) students' perceptions on the content of the ELMs, and the perceived influence of the ELMs on students' performance during clinical rotations. SUBJECTS AND METHODS: All CT students watched nine ELMs before the related classroom lecture and group discussion. Following that, students completed nine preclinical rotation surveys. After their clinical rotations, students completed nine postclinical rotation surveys. RESULTS: Statements on the content of the ELMs regarding the quality of the video and audio, duration, navigation, and the materials presented, received positive responses from the majority of the students. While there were a few disagreements and neutral responses, most of the students responded positively saying that the ELMs better prepared them for their role, as well as helped them to better perform their roles during the clinical rotation. The majority of the students recommended developing more EMLs for cytology courses in the future. CONCLUSIONS: This study has given hope that the ELMs have potential to enhance our online curriculum and benefit students, within the United States and internationally, who have no easy access to cytology clinical laboratories for hands-on training.
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INTRODUCTION: Sustaining a well-trained, competitive, and marketable cytotechnology workforce is a major goal of the American Society of Cytopathology/American Society for Clinical Pathology Workgroup Focusing on Emerging Roles in Cytopathology (ASC/ASCP Workgroup). This article describes the data collection initiatives performed by the Workgroup to confirm cytotechnologist (CT) perceived educational needs to keep pace with emerging practice changes in the workplace. In response to data collected, the Workgroup created Advanced Cytopathology Education (ACE)-a unique, innovative educational resource designed to fulfill this need. MATERIALS AND METHODS: Data collection efforts included annual needs assessment surveys to gauge CT participants' preferred topics and learning modalities. These data were used to design and prioritize ACE topics. Program evaluations were also administered to evaluate participants' perceptions of program quality and effectiveness at meeting their educational needs. RESULTS: Research findings reveal CT education needs to align with emerging practice areas as reported in other Workgroup data collection efforts. The incorporation of new entry-level competencies in cytotechnology training programs prepares new CT graduates, but there is no standardized mechanism for formal, robust, and recognized ongoing education for other practicing CTs. CONCLUSIONS: This article highlights the educational endeavors carried out by the Workgroup in fulfilling the educational needs of practicing CTs as communicated through various data collection efforts. As CT responsibilities evolve, ASC and Workgroup efforts will continue to focus on providing educational support for current practicing CTs who need training in emerging practice areas.
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OBJECTIVE: The aim of this work was to raise awareness of problems using digital applications for examining, teaching, and applying telecytology at King Abdulaziz Medical City (KAMC), Riyadh, Saudi Arabia; University of Nebraska Medical Center (UNMC), Omaha, NE, USA; and University of Pittsburgh Medical Center (UPMC), Pittsburgh, PA, USA. The objective was to rationalize problems and propose alternative digital approaches. STUDY DESIGN: We sought to identify solutions to improve the following: (a) interpretive examination scores at KAMC for complex cytological templates (i.e., high-grade squamous intraepithelial lesions [HSIL]) when using static digital images (SDI) of cells in regions of interest (ROI); (b) visualization of cells in 3D clusters when teaching at UNMC using 2D and 3D whole-slide imaging (WSI); and (c) visualization of cells through streaming telecytology at UPMC. RESULTS: Composite SDI (CSDI) improved test scores for complex interpretations (i.e., HSIL) by converging diagnostic criteria from multiple ROI. Multiplane focusing through z-stacked WSI facilitated the teaching of cytological entities characterized by 3D cell clusters and consultative telecytology through robotic cell analysis. CONCLUSIONS: Adequately visualized cytomorphology and multiplane focusing are essential for virtual cytopathology examinations, teaching, or consultative telecytology. Visualization of diagnostic criteria through 2D or 3D imaging is critical. Panoptiq panoramic WSI with integrated z-stacked video clips enables optimal applied telecytology.
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Instrução por Computador , Educação Médica/métodos , Interpretação de Imagem Assistida por Computador/métodos , Microscopia/métodos , Patologia/educação , Lesões Intraepiteliais Escamosas Cervicais/patologia , Telepatologia/métodos , Neoplasias do Colo do Útero/patologia , Feminino , Humanos , Nebraska , Teste de Papanicolaou , Pennsylvania , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Arábia Saudita , Esfregaço VaginalRESUMO
BACKGROUND: Interprofessional education (IPE) is becoming increasingly prevalent in health science education, with the goal of preparing students to work collaboratively in teams within the healthcare environment. Students in our cytotechnology and radiation therapy (RT) programs used virtual technologies to demonstrate their professions using case studies. The purpose of this activity was to see if our students' knowledge of each other's professions and educational technologies increased and if the students had a better understanding of how they would work together in a healthcare team. METHODS: Participants included four cytotechnology students and five RT students. All were given a presurvey to determine their level of knowledge about each other's profession. The cytotechnology students presented cases involving gynecologic and lung cancers using virtual microscopy and explained how they screen slides and interpret cellular changes. The RT students explained how they would treat these same patients using the Virtual Environment Radiotherapy Training system (VERT), showing the cytotechnology students how the beam is guided to the exact spot for treatment. After the IPE activity, all participants were given a post-survey to determine their levels of understanding. RESULTS: The results indicated that the IPE activity increased the level of understanding regarding each other's professions and how they each fit together in the role of patient care. CONCLUSIONS: IPE activities, even on a small scale with two professions in the same college, can improve knowledge and collaboration between professions. More of these activities should be conducted for effective healthcare teams and improved patient outcomes.
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Ocupações Relacionadas com Saúde/educação , Relações Interprofissionais , Equipe de Assistência ao Paciente/organização & administração , Realidade Virtual , Atitude do Pessoal de Saúde , Comportamento Cooperativo , Humanos , Conhecimento , Papel ProfissionalRESUMO
OBJECTIVE: Cytodiagnoses of specific malignancies are enabled through analyses of abnormal nuclear chromatin and cytoplasmic features in stained cells. AIM: The objective of this work was to explore the inception, development, and chemistry of the Pap stain method introduced in 1942 by Dr. G.N. Papanicolaou. STUDY DESIGN: To achieve this, we carried out a review of the English literature. RESULTS: Between 1914 and 1933, Papanicolaou first analyzed vaginal squamous cells in guinea pigs and later in human vaginal fluid samples using hematoxylin and eosin with limited color reactions, correlating the cell-type morphology with endocrinology and histology. The 5-dye Pap stain method evolved through 2 salient phases. The first, between 1933 and 1942, saw the introduction of alcohol-ether fixation and aqueous waterblue staining to enhance cellular transparency, aiding the distinction of cervical cancer cells from benign cells, with quantitative and qualitative assessment of squamous cell maturity. The second phase, between 1942 and 1960, saw the introduction and refinement of various alcoholic cytoplasmic counterstaining schemes with orange G and EA (light green, Bismarck brown, eosin) and phosphotungstic acid, allowing wider ranges of polychromasia and further enhancing cellular visualization, facilitating the distinction of cell types and improving diagnostic confidence. CONCLUSIONS: Development of the Pap stain method followed specific historical and scientific events. The staining method evolved following incremental improvements in cellular transparency achieved through tailored cellular fixation and cytoplasmic staining using variable dye and pH combinations.
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Corantes/química , Teste de Papanicolaou/métodos , Esfregaço Vaginal/métodos , Animais , Citoplasma/patologia , Feminino , Humanos , Coloração e Rotulagem/métodos , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/patologiaRESUMO
BACKGROUND: Our cytotechnology (CT) program has been utilizing virtual microscopy (VM) as an adjunct educational resource since 2011. AIMS: The aim of this study was to identify the utilization of VM in other CT programs across the United States (US). SUBJECTS AND METHODS: A cover letter was sent to the program directors of all accredited CT programs in the US (excluding our program), requesting their participation in an online survey. After 2 days, the participants were sent an online link to the survey. The survey results were analyzed using descriptive statistics. RESULTS: There were a total of 25 respondents to the survey. Among the 25, three CT programs use VM. Two of the three programs have been using VM for <2 years while another program for "2-4" years. The respondents found that VM's side-by-side comparison feature helped to demonstrate differences between diagnoses and preparation methods, and VM helped to preserve the important slides by digitizing them. Respondents believed that teaching with glass slides was very important. The reasons for not using VM were that VM is expensive and time-consuming to incorporate into the program, and lack of manpower resources to create digitized teaching files. CONCLUSIONS: The CT programs that use VM found it to be a valuable educational tool. Even though many were not using VM, responses from the survey indicated they will likely use it in the future.