bluuebunny/crossref_metadata_2025 huggingface dataset:
Sialic (Neuraminic) Acid Residues Detection in Pulmonary Adenocarcinomas: A Digital Assessment
Authors: MARIASSY ANDREW T , DANIHEL LUDOVIT , WEISMANN PETER , DRIBIN LORI , KUBIKOVA ELISKA , BABAL PAVEL
Published: June 2021

Glycocalyx on the cell surfaces interacts with adjacent cells and the endothelium. Neoplastic cellular changes are known to affect the glycocalyx and the “stickiness” of the cell surface. We hypothesize that the sialyl residue expression is differentially altered in pulmonary adenocarcinoma and this change affects their invasiveness. In this study, we quantitatively assessed sialic acid residues in pulmonary adenocarcinomas by utilizing a sialyl recognizing Trithrichomonas mobililensis lectin (TML). Biotinylated lectin binding was detected in 24 subjects. The TML reacted paraffin sections of resected pulmonary adenonocarcinomas, were digitized at constant illumination with IMT4 CCD Camera and the images were morphometrically evaluated with the Olympus Analysis Program. For more precise estimation of the reaction product on TML, we reduced the blue signal and accentuated the red one in all images equally with ADOBE Photo Shop Elements 2.0. The edited images had a sharper definition of the reaction product and allowed a more precise measurement of the sialic acid residues in the area of interest after subtraction of airspaces background from the measured tissue area. Detected sialic acid ranged from 0.5% to 3.3% of the examined tissue. Pulmonary adenocarcinomas are recognized as having considerable pleomorphism, as well as mixture of subtypes. We examined the relationship of sialic acid abundance with histopathologic tumor pattern and TNM (Tumor, Nodes, Metastasis) classification. The early stages of tumors, having the lepidic, papillary and acinar patterns in stages 0, I & II with no metastasis, tended to have high, >.5% of detected sialyl residues. The low, <0.5% expression of sialic acid was found to be correlated with the invasive subtypes; the micropapillary, mixed and solid patterns in stages III and IV. These results indicate that the sialyl residue abundance was inversely correlated with the invasiveness and the stage of adenocarcinoma. We conclude that the sialic acid residues as detected with the TML, maybe an important determinant of the tumor invasiveness and an adjuvant for a more objective determination of the cell glycocalyx transformation for staging the pulmonary adenocarcinoma.

Support or Funding Information

NSU Faculty Research Grant.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Meshes, Molecules and Mice: Methods in Multi‐user Morphometry
Authors: Lozanoff Scott
Published: June 2021

Quantitative analysis of anatomical landmark data provides insight into the functional relationship between phenotypic and genotypic variability. Morphological meshes formed by bounding anatomical landmarks can be generated and facilitate testable hypotheses concerning underlying biological mechanisms driving regional development. Early computer systems limited multi‐landmark analysis; however, recent advances enable surface mesh extrapolation from volumetric models and quantitative comparison providing further resolution of cellular activities regulating morphogenesis. Although current technology enables image analysis using augmented reality devices, remote collaborative interaction remains limited. The purpose of this presentation is to review examples of quantitative phenotypic descriptions associated with the identification of gene defects resulting in craniofacial malformations in experimental mouse models and to describe new software that provides web‐based remote access and collaborative interaction of volumetric models and morphometric analysis. Utilizing two experimental mouse mutation models (Br and tuft), landmark‐based morphometric analysis revealed defective regional development during craniofacial morphogenesis. Gene mapping analysis showed complicated defects involving the six2/six3 locus (Br) and TET1 (tuft). Subsequent molecular expression analysis (IHC, WMISH, qPCR, RNA seq) within morphometrically defined regions identified underlying morphogenetic defects. A web‐based 3d imaging tool (Rad3d) was developed that enabled real‐time editing and analysis of volumetric data. Given the need for investigator collaboration, we provided a platform for multi‐user real‐time editing, security, and storage for users. This web application enables multi‐user access and real‐time interaction with 3d volume data essential for morphometric analysis.

Support or Funding Information

R01DK064752, HRSA‐OAT, AHA‐9951133Z, XLR8

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Why do Animals get Sick? Evolutionary Veterinary Medicine: The Benefits of Expanding from a Comparative to an Evolutionary Perspective
Authors: Uhl Elizabeth W
Published: June 2021

Veterinary medicine is comparative, but, as in human medicine, the evolutionary origins of disease susceptibility are not fully considered. Thus while there are good descriptive studies, many ‘why’ questions concerning disease remain unanswered, and many are not even asked. Since disease in every animal species except humans falls under the purview of veterinary medicine, this focus on descriptive ‘how/what’ mechanisms combined with breakthroughs in technology has led to a rapidly expanding knowledge base that is impossible for veterinarians and researchers to keep up with and is overwhelming students. Consideration of evolutionary heritage in veterinary basic teaching/research and clinical case work ups provides a much better and more universal basis for understanding disease than that provided by explanations based only on clinical signs, morphology and molecular mechanisms. For example: evolved structural and functional adaptations for movement and to diet often determine disease susceptibilities across species as well as the effectiveness and toxicity of drugs. Similarly, while specific pathogens vary in their molecular mechanisms of infection, they often exploit the same vulnerabilities as to cell type infected and the features of diseases induced across species. Questions of why disease occurs in specific locations can be understood and further investigated through consideration of organ evolution: for example why vascular tumors arise on the right side of the heart in both dogs and people. Diseases induced through environmental mismatches (e.g.: hemochromatosis), toxins (e.g.: cancer) or changes in microbiome (e.g.: allergic/autoimmune diseases) reveal common shared susceptibility and pathology across species. At the molecular level, understanding the evolutionary basis for why there are both species similarities and differences in disease induced by defects in the same conserved gene (e.g.: dystrophin), can inform treatment options. Adding an evolutionary perspective to veterinary medicine provides a context for organizing and interpreting the ever‐increasing amount of data being generated and allows the critical ‘why’ questions, which are unable to be answered through descriptive studies, to be investigated. In addition, the very close connections between human and animal health (One Health) can only be understood in light of evolution.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Synthetic evolution of C4 photosynthesis
Authors: Weber Andreas PM
Published: June 2021

C4 photosynthesis has independently and convergently evolved approx. 70 times in flowering plants. Most C4 plant species display a distinct leaf anatomy (Kranz‐anatomy) and similar metabolic and biochemical features. Computational modeling indicates that C4 photosynthesis evolves from C3 photosynthesis via C3–C4 intermediate states [1,2]. C3–C4 intermediacy evolves via simple loss of function mutations and causes a nitrogen disbalance between leaf mesophyll and bundle sheath cells. Overcoming this nitrogen disbalance requires the shuttling of organic acids between mesophyll and bundle sheath cells, which under permissive conditions drives the system towards C4photosynthesis [3]. Genetic hybrids between C3 and C3–C4 intermediate species segregate for subtraits of C3–C4 intermediacy and thereby enable the identification of key enablers by quantitative genetics. Identification of such enables is a prerequisite for engineering of the C4trait into C3species by synthetic biology approaches [4].

We have recently identified such enablers. These include cis‐regulatory elements in the promoters of genes encoding components of photorespiration. To experimentally test these enablers in a C3 genetic background and to eventually recapitulate the evolutionary trajectory from C3to C4photosynthesis in the model plant A. thaliana, we have developed a non‐invasive visual marker that enables detection of Cas9‐activity in planta [5,6]. We have used this maker system to test different concepts for homology‐based gene editing and have developed a reliable system for homology‐directed repair in Arabidopsis [7]. These protocol will now be employed for introduction of C4‐enablers into Arabidopsis.

Support or Funding Information

We appreciate support by the Deutsche Forschungsgemeinschaft (EXC 1028, IRTG 1525, ERA‐CAPS project C4BREED).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Circadian biology in translation
Authors: Hogenesch John
Published: June 2021

The past several decades has seen an explosion of growth in mechanistic understanding of circadian clocks in several model organisms, including humans. However, translation of that knowledge into actionable medical intervention(s) has been slow to non‐existent. Here, I will discuss some of the reasons why this is as well as our efforts to mitigate this using systems biology approaches. I will talk about our recent progress in understanding the molecular output of the clock in the mouse and humans, including identifying new opportunities for circadian dosing time in improving drug action ‐‐ hypothesis‐driven, mechanistic circadian medicine. I will talk about our efforts to test these hypothesis prospectively in model organisms and retrospectively in large clinical databases. Finally, I will discuss future opportunities and challenges.

Support or Funding Information

This work is supported by the National Institute of Neurological Disorders and Stroke (2R01NS054794 to JBH and Andrew Liu; and 1R21NS101983 to Tom Kilduff and JBH), the National Heart, Lung, and Blood Institute (R01HL138551 to Eric Bittman and JBH), and National Human Genome Research Institute (2R01HG005220 to Rafa Irizarry and JBH.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Responses to replication stress in human cells
Authors: Bielinsky Anja‐Katrin , Leung Wendy , Shetty Mihir , Starr Tim , Moldovan George‐Lucian
Published: June 2021

Genome integrity relies on faithful genome inheritance. DNA damage induced replication stress leads to the generation of single‐stranded (ss) DNA, which triggers the ubiquitination (Ub) of the replication clamp, proliferating cell nuclear antigen (PCNA) at lysine 164 (K164). This activates two DNA damage tolerance (DDT) pathways: 1) error‐prone translesion synthesis, or 2) error‐free template switching. We generated PCNAK164R knock‐in cell lines via CRISPR/Cas9‐mediated gene targeting in HCT116, a colorectal cancer cell line and hTERT‐RPE1, an immortalized nontransformed epithelial cell line. RPE1 PCNAK164R hemizygous and homozygous mutants showed an increase in fork stalling and a decrease in new origin firing under unperturbed conditions. Moreover, these cells also exhibited accelerated fork speed. Although homologous recombination was enhanced in the mutants, it was not further upregulated in the presence of genotoxic drugs. We are currently testing the replication dynamics of these RPE1 mutants in response to DNA damaging agents. We hypothesized that cancer cells are more dependent on PCNA K164 for survival than nontransformed cells. Indeed, we were unable to isolate homozygous cell lines in HCT116. Heterozygous PCNAK164R/+ mutants displayed a slow‐growth phenotype, increased DNA damage sensitivity and chromosome breaks following UV‐light exposure, indicating a reduced capacity to tolerate extrinsic replication stress. These mutants also exhibited massive telomere erosion that was not caused by a decrease in telomerase activity, suggesting that PCNA‐K164 is essential in HCT116 because it regulates telomere length. In contrast, in a HCT116 PCNAK164R/+ mutant xenograft mouse model tumor burden was increased compared to wildtype. Thus, the slow‐growth phenotype in vitro is not predictive of the growth behavior in vivo. RNA sequencing revealed that tumor tissues arising from PCNAK164R/+ mutant cells drastically downregulated several DNA repair pathways, providing a mechanistic explanation for the enhanced tumor burden. Together, our data argue that DDT pathways are important for preserving genome stability and protecting against tumor growth.

Support or Funding Information

Supported by NIH R01 GM074917 (AKB).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Metabolic Origin of the Fused Aminoacyl tRNA Synthetase, Glutamyl‐Prolyl tRNA Synthetase (EPRS)
Authors: Eswarappa Sandeep M. , Potdar Alka A. , Sahoo Sarthak , Sankar Santhosh , Fox Paul L.
Published: June 2021

About 1 billion years ago, in a single‐celled holozoan ancestor of all animals, a gene fusion of two tRNA synthetases formed the bifunctional enzyme, glutamyl‐prolyl tRNA synthetase (EPRS). We propose here that a confluence of metabolic, biochemical, and environmental factors contributed to the specific fusion of glutamyl‐ (ERS) and prolyl‐ (PRS) tRNA synthetases. To test this idea we developed a mathematical model that centers on the precursor‐product relationship of glutamic acid and proline, as well as metabolic constraints on free glutamic acid availability near the time of the fusion event. Our findings indicate that proline content increased in the proteome during the emergence of animals, thereby increasing demand for free proline. Together, these constraints contributed to a marked cellular depletion of glutamic acid and its products, with potentially catastrophic consequences. In response, an ancient organism invented an elegant solution in which genes encoding ERS and PRS fused to form EPRS, forcing co‐expression of the two enzymes, and preventing lethal dysregulation. The substantial evolutionary advantage of this co‐regulatory mechanism is evidenced by the persistence of EPRS in nearly all extant animals.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Elucidation of the Pax‐5/miRNA interactome and its oncogenic effects in cancer cells
Authors: Robichaud Gilles Andre , Harquail Jason , Beauregard Annie‐Pier , Hannay Brandon
Published: June 2021

The Pax‐5 oncogene has consistently been associated to B cell cancer lesions and more recently solid tumors including breast carcinoma. Although Pax‐5 downstream activity is relatively well characterized, aberrant Pax‐5 expression in a cancer specific context is poorly understood. To investigate the regulation of Pax‐5 expression, we studied micro‐RNAs (miRNAs) and mRNA polymorphism in cancer cells. Using bioinformatics and next‐generation sequencing, we found that miRNAs 484 and 210 are aberrantly expressed in breast cancer and predicted to target Pax‐5 mRNA. Through conditional modulation of these miRNAs, we demonstrate that miRNA‐484 and 210 inhibit Pax‐5 expression and regulate Pax‐5‐associated cancer processes. In validation, we show that these effects are reversible by either Pax‐5 recombinant overexpression; or, selective miRNA inhibition. On the other hand, upon our analysis of the Pax‐5 transcript, we found that not only is the 3′UTR submitted to alternative polyadenylation; but also, alternative splicing of the Pax‐5 mRNA 3′UTR. These transcript polymorphism events lead to the shortening of the Pax‐5 3′UTR which resulted in greater oncogene translation frequency. More importantly, we also found that Pax‐5 transcripts characterized with shorter 3′UTRs were associated with advanced staging of cancer lesions. Our findings identify novel molecular mechanisms, which account for Pax‐5 aberrant expression and function in cancer cells. These findings will further elucidate Pax‐5‐mediated cancer processes and may provide new avenues for therapeutic intervention.

Support or Funding Information

This work was supported by grants from the New Brunswick (NB) Innovation Foundation, the Canadian Breast Cancer Foundation, the Canadian Breast Cancer Society/QEII Foundation, the NB Health Research Foundation and by the Beatrice Hunter Cancer Research Institute

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Interleukin‐10 Contributes to Macrophage Dysfunction in Acetaminophen‐induced Acute Liver Failure
Authors: Pant Asmita , Roth Katherine , Kopec Anna , Luyendyk James , Copple Bryan
Published: June 2021

Acetaminophen overdose is the leading cause of acute liver failure in the United States. In mice, interleukin‐10 (IL‐10) deficiency dramatically increased early inflammation and hepatotoxicity after acetaminophen challenge. Paradoxically, IL‐10 levels are highest in acute liver failure patients with the poorest prognosis. The mechanistic basis for this apparent discrepancy is not understood. We examined the function of IL‐10 in a mouse model that recapitulates failed liver repair analogous to acute liver failure in patients, wherein mice are treated with a large dose of acetaminophen (i.e., 600 mg/kg). Hepatic IL‐10 mRNA expression was dramatically increased 24 hours after challenge of mice with 600 mg/kg APAP, whereas IL‐10 induction was not observed at this time point in mice given 300 mg/kg APAP. At 24 hours, few IL‐10 expressing cells were detected in the livers of mice challenged with 300 mg/kg acetaminophen, whereas numerous macrophages and other hepatic cells expressed IL‐10 in mice treated with 600 mg/kg acetaminophen. Necrotic lesions were effectively cleared in mice given 300 mg/kg APAP, whereas inefficient macrophage trafficking into the necrotic lesions was associated with failed repair in mice given 600 mg/kg APAP. Remarkably, administration of an IL‐10 neutralizing antibody, 24 hours after treatment of mice with 600 mg/kg APAP, fully restored macrophage accumulation in areas of hepatic necrosis, suggesting exaggerated IL‐10 expression causes macrophage dysfunction essential for repair. Consistent with this hypothesis, administration of recombinant IL‐10 to mice with established APAP (300 mg/kg)‐induced liver necrosis delayed liver repair alongside a marked reduction in macrophage infiltration. Collectively, these studies indicate that dysregulation of IL‐10 in the late repair phase after acute liver injury impairs intrahepatic macrophage trafficking, which may explain failed liver repair and a poor outcome in acute liver failure patients.

Support or Funding Information

Supported by NIH Grant DK103895 to BLC and NIH Grant DK105099 to JPL.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Stimulator of Interferon Genes Deletion in Primary Heart Endothelial Cells Results in Decreased T Cell Transmigration
Authors: Anastasiou Marina , Carrillo‐Salinas Francisco J. , Kaur Kuljeet , Muendlein Hayley , Poltorak Alexander , Alcaide Pilar
Published: June 2021

The vascular endothelium is a major regulator of immune responses following injury and organ specific inflammation, in part by actively participating in adhesion and subsequent transendothelial migration (TEM) of leukocytes from the blood stream into tissues. Stimulator of Interferon genes (STING) acts as a cytosolic DNA sensor as well as an adaptor molecule for type I interferon signaling. STING is also an important regulator of paracrine inflammatory responses, effectively functioning as a “signaling hub” molecule. Our current data shows that primary mouse heart endothelial cells (MHEC) express STING, and we tested the hypothesis that heart endothelial STING modulates leukocyte adhesion and/or TEM. C57BL/6 WT and STING‐deficient MHEC were isolated from mouse hearts using positive selection and were grown to confluency in vitro. Upon confluency the monolayers were stimulated with TNFα to induce adhesion molecule upregulation. Activated Th1 cells were generated in vitro by positively selecting CD4+ cells from WT spleen and inguinal lymph nodes and stimulating them with a Th1‐specific cocktail. Th1 cells were then perfused across TNFα‐activated primary MHEC. Adhesion, TEM, and total accumulated cells (adhered plus TEM) under defined laminar flow conditions were analyzed by real time videomicroscopy using a parallel plate flow chamber. STING‐deficiency in MHEC resulted in decreased T cell TEM, even in the presence of strong chemokine stimulation but did not alter accumulation. This phenotype correlated with marginally decreased expression of ICAM‐1 in the STING‐deficient TNFα‐treated MHEC. RNA‐sequencing analysis of WT and STING‐deficient MHEC indicated further differences between the groups, particularly in respect to immune‐related genes and genes coding for endothelial cell adhesion molecules that play a role in leukocyte recruitment and TEM. Ongoing studies will determine the contribution of heart endothelial STING to leukocyte recruitment and TEM in vivo and dissect the mechanisms regulating the heart endothelial STING‐mediated innate sensing upon inflammation.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
FPR2 regulates monocyte recruitment to promote mucosal wound repair
Authors: O'Leary Monique N. , Birkl Dorothee , Quiros Miguel , Reed Michelle , Azcutia Veronica , Schaller Matthew , Nishio Hikaru , Keeney Justin , Lukacs Nicholas W. , Neish Andrew S. , Parkos Charles A. , Nusrat Asma
Published: June 2021

Mucosal wound repair is coordinated by a dynamic spatiotemporal cross talk of mediators with receptors on epithelial cells and infiltrating immune cells. After initial recruitment of neutrophils, monocytes infiltrate mucosal wounds in close proximity to repairing epithelium. Key regulators of mucosal repair include pro‐inflammatory and pro‐resolving mediators that help orchestrate temporal recruitment of immune cells, resolve acute inflammatory responses and regulate epithelial cell migration and proliferation. Immune cell derived mediators have been shown to activate G‐protein coupled receptors (GPCR) that significantly impact the inflammatory response and repair. In this report, our studies were focused on how a GPCR, formyl peptide receptor 2 (FPR2), orchestrates mucosal repair. Fpr2/3−/− mice were utilized to investigate the role of formyl‐peptide receptor 2/3 (FPR2/3), an orthologue of human FPR2/ALX, in regulating intestinal mucosal repair. Compared to wild type mice, Fpr2/3−/− mice exhibited delayed recovery from acute colitis and impaired repair after biopsy induced injury. Time course analyses of immune cell populations in the wound bed of wild type and Fpr2/3−/− mice by flow cytometry revealed mice lacking FPR2 have a reduced influx of inflammatory monocytes (defined as CD45+ CD11b+ Ly6G− Siglec F− F4/80lo Ly6Chi) on day 1 and 2 post wounding compared to wild type mice. To further supporting these data, we assessed expression of CCR2, a chemokine receptor and marker of conventional Ly6Chi inflammatory monocytes, in wounded tissue. We found reduced expression of CCR2 mRNA in tissue isolated from Fpr2/3−/− mice. Bone marrow transplant experiments revealed that monocytes from wild type mice had a competitive advantage over Fpr2/3−/− cells in migrating into healing mucosal wounds. Moreover, Fpr2/3−/− monocytes were defective in chemotaxis responses to the chemokine CCL20, a chemoattractant for monocytes that is upregulated during the early phase of inflammation. Analysis of Fpr2/3−/− monocytes revealed altered expression of the CCL20 receptor, CCR6, suggesting a role of Fpr2/3 and CCL20‐CCR6 signaling in regulating monocyte recruitment to sites of mucosal injury. In summary, these findings identify the contribution of monocyte expressed FPR2/3 in facilitating their recruitment to sites of mucosal injury to promote wound repair.

Support or Funding Information

This research was funded by DK055679, DK089763, DK059888, to AN; DK61739, DK72564, DK79392, to CAP; and a German Research Foundation Research Fellowship (SI 2282/1‐1, to DB).

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Dynamics of antibody‐virion complexes reveals contrasting stabilizing and destabilizing effects of neutralizing antibodies on dengue
Authors: Anand Ganesh , Lim Xin Xiang
Published: June 2021

Dengue viruses are dynamic macromolecular assemblies in solution that stably package genomic RNA outside the human host but are programmed to undergo timed disassembly under opportune host environments. This inherent metastability is manifested as reversible conformational rearrangements referred to as ‘breathing’ and are critical for sensing host‐specific environmental cues, receptor interactions and for initiating host entry. The virion envelope proteins are the primary targets for antibodies. How only a subset of binding antibodies are capable of neutralizing dengue is an open question and represents a major challenge in therapeutic antibody discovery and vaccine design to tackle dengue. The key to understanding how classes of neutralizing antibodies work is inherently linked to the inherent dynamics of whole viral particles in solution. Amide hydrogen deuterium exchange mass spectrometry (HDXMS) is a powerful tool for measuring hydrogen bonding propensities and solvent accessibility of proteins and protein complexes. This makes it especially suitable for probing the breathing dynamics, measurement of quaternary contacts of viral particles in solution as well as to probe the effects of host‐specific conditions and for mapping epitopes and paratopes of neutralizing antibodies. We have carried out dynamics measurements on two classes of neutralizing antibodies‐ 2D22 and C10 with the same DENV2 NGC strain. Our HDXMS results shows that 2D22 binding results in decreased deuterium exchange while C10 binding enhances deuterium exchange across the dengue virion. Viruses are intrinsically metastable and maintained in partially stable assemblies. Our results describe contrasting modes of antibody neutralization. One where antibody binding impairs the intrinsic breathing. We describe for the first time, how C10 binding destabilizes the virus to preemptively disassemble it prior to reaching its target cells. These unique antibody effects and serotype dynamic profiles also show that the epitope landscape across the viral surface is non‐uniform and symmetry unit‐specific. Preferential antibody binding to symmetry units on the icosahedral viral surface further underscores the importance of quaternary interactions in efficacy of antibody action, with major implications for antibody‐dependent enhancement of dengue.

Support or Funding Information

Competitive Research Proposal (CRP)‐ National Research Foundation (NRF), Singapore

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Should Black Humor Be Put To Death? Examining Student and Staff Views on Black Humor in Anatomy Labs
Authors: Duenas Angelique , Kirkness Karen , Finn Gabrielle
Published: June 2021

Beyond studies and discussions regarding learning outcomes related to dissection, prosection, and anatomy lab experiences, there has been a recent increase in examining the role of the ‘hidden curriculum’ in anatomy education. Lab experiences of many types can help foster professionalism and more humanistic views towards donors, and eventually patients, via this integrated hidden curriculum. But what has been overlooked are the potential darker aspects of the hidden curriculum that may begin in anatomy labs. While the days of ‘cadaver antics' and directly objectifying donor bodies are no longer tolerated, there is little research regarding the more subtle use of dark humour and language still present in anatomy education. Thus, our study aims to better understand the use of black, or cynical, humour as a component of the hidden curriculum of anatomy labs. An Ethics‐reviewed study was designed, and participants were recruited from the Hull York Medical School to voluntarily complete an online survey comprised of 3 demographic, 5 open‐ended, and 6 rating‐scale items. Eligible participants included medical students, MSc anatomy students, anatomy lab tutors, and anatomy faculty. Responses were analysed by descriptive statistics, and open‐ended questions were coded for common themes. Data continues to be collected and analysed, but preliminary findings suggests there are certain themes regarding the perceived appropriateness of black humour in medical education settings. Our findings suggest that the themes of context and situation of humour are key in determining appropriateness, and that while some participants express discomfort at the use of black humour, humour in the anatomy lab is more widely regarded as an appropriate means of coping with stress. An additional emergent theme is the use of (cynical) humour to promote camaraderie amongst teammates. While our data analysis is still ongoing, the preliminary themes suggest outcomes of this study may have implications for better understanding how to address the darker hidden curriculum of anatomy. Additionally, we hope that our findings help to further develop best practices for fostering humanistic future practitioners, throughout all levels of medical training, but starting in anatomy labs with students' ‘first patients.’

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
A Digital Anatomico‐Surgical Module for Preparing the 21st Century Dermatologist
Authors: Adar Tony , Pagano Anthony S , Monheit Gary D , Marquez Samuel
Published: June 2021

Strong anatomical acumen is critical in today's procedural dermatologic practice. Accruing the necessary skill set for the dermatologic resident to become a successful practitioner requires acquiring the foundational knowledge of the underlying craniofacial anatomy with the goal of facilitating and translating basic principles of cutaneous surgical procedures. Most dermatologic residency programs provide hands‐on surgical experience supplemented with academic lectures and surgical texts. In response to growing demand from the dermatology residents at SUNY Downstate Medical Center the co‐investigators recently combined efforts to organize and instruct a cadaveric head and neck dissection course with photo documentation. Feedback from residents strongly advocated for dissection sequence to be video captured in order to re‐visit protocol, assess understanding and implement practice procedures. This study reports on the creation of a digitally annotated facial dissection high definition video with clinically relevant correlates providing a comprehensive review and enhancing anatomical education to dermatology residents. Specifically, the dermatologic resident was guided through a complete fascial dissection sequence to delineate key anatomical landmarks, detailing facial planes and fat compartments pertinent to dermatologic procedures and surgeries. Pre‐ and Post‐assessment tests provided an evaluation of the impact the digital educational material had on the dermatologic residents. Resident perception reflected an increase in confidence and understanding of the surgical approaches required for successful outcomes. Video distribution to residents would permit mastery of craniofacial anatomy, which remains indispensable for proper and safe cutaneous and cosmetic procedures for the 21st century dermatologist.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Visualization of Heart Electrocardiographic Activity using a Mixed Reality System
Authors: Michalski Connor , Cowan Melissa , Bohinsky Jonathan , Dickerson Ryan , James Bradford , Plochocki Jeffrey H
Published: June 2021

The use of three‐dimensional (3D) visualization technology in medical education and physician training has been demonstrated to improve learning outcomes. One emerging visualization tool that is being explored to supplement anatomy and physiology education is mixed reality, a technology that overlays virtual objects into the real world. The challenge faced by mixed reality is creating applications that are user friendly and have educational value beyond the static exhibition or manipulation of anatomic representations. We developed an application that uses the HoloLens™ (Microsoft Corporation, Redmond, WA, USA), a wearable mixed reality system, to integrate 3D animations of heart anatomy with electrocardiography. The application was created by UCF College of Medicine in Unity v.2017.3.1f1 (Unity Technologies, San Francisco, CA, USA) using the Microsoft HoloToolkit v.2017.4.2.0 and Vuforia Engine v.7.0.47 (PTC, Needham, MA, USA). Using this application, the HoloLens scans a QR code corresponding to an electrocardiogram (ECG) and generates a 3D animation of heart contractions that corresponds to the electrical activity represented in the ECG. Gesture recognition is used as an input mechanism to encourage engagement and interactions with the 3D heart models. Audio narrations are used to describe the context of each animation and learners can test their knowledge using the built‐in formative quiz. Our mixed reality application allows students to visualize cardiac functional morphology in 3D space. It facilitates training physicians to conceptualize the anatomy of sinus rhythm and cardiac arrhythmias, and transfer their knowledge of myocardial depolarization and repolarization to realistic clinical scenarios. It accomplishes these educational goals in a manner that cannot be accomplished with traditional visualization technologies or anatomic models. Future development of our application will focus on adding displays of hemodynamics, including pressures of the heart chambers and great vessels and corresponding disturbances in cardiac blood flow.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Pocket Pelvis: Developing an Augmented Reality App for Better Understanding of Pelvis Anatomy
Authors: Dixit Ishan , Prakitpong Ratthamnoon , Blumer Paige , He Ellen , Jeong Eric , Kim Jason , Oh Jueyong , Ghomi Mehrdad , Krebs Claudia , Cerron Dante
Published: June 2021

The human pelvis is one of the more complicated anatomical regions for students to understand due to its complex 3D geometry, the layered pelvic walls, and the differences between male and female pelvises.

In order to facilitate the 3D learning of the pelvis we developed an augmented reality application that allows the learner to visualize structures of the pelvis on a 3D printed pelvis model. The 3D printed pelvis model was created based on volumetric reconstruction of the NIH Visible Human dataset. Pelvis structures, such as muscles and neurovasculature, were created using Blender 3D modeling software. The application was created using Unity, and Vuforia was used for the augmented reality image recognition. The app was programmed for use on Android or iOS devices.

The aim of this app was not to re‐create an anatomical model, but rather to create a teaching tool that is both accurate and conceptual in nature. Iterative design of the user interface and the user experience led to an educational experience where students can manipulate the 3D printed pocket‐sized pelvis and have control over which structures they can view in the augmented reality app. This approach has led to a set of best practices for this type of augmented reality app: The 3D printed object needs to be generated using a filament that is not too shiny to minimize glare, the tags recognized by Vuforia for adding the augmented reality structures need to be unique and their size needs to be a compromise between optimal recognition and non‐interference with the anatomy. The user interface needs to be clearly structured and allow the user to interact with the augmented reality structures and their device seamlessly and simultaneously. The combination of a physical object that can be freely turned with augmented reality structures that can be turned on and off by the user leads to a user experience that combines a haptic experience with visual cues – a combination that is a foundation for a deep anatomical understanding of the pelvis.

Support or Funding Information

Supported by the Strategic Investment Fund from the UBC Faculty of Medicine and the UBC Teaching and Learning Enhancement Fund

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Facial nerve morphogenesis: three‐dimensional and multiplanar study
Authors: Labrousse Marc , Dubernard Xavier , Settembre Nicla , Micard Emilien , Braun Marc , Renard Yohann
Published: June 2021

The purpose of this study was to describe the facial nerve embryology, using three‐dimensional reconstructions from human embryos histological sections, and to compare our results with literature. Human embryos stage 13 to 21 from Carnegie collection and Paris‐Descartes University's Delmas‐Rouvière collection stained with hematoxylin & eosin or Masson's trichrome were used. A local made computer software developed by IADI lab (INSERM U947, Nancy) allowed automatic slices selection, automatic rigid registration, and dust filtering. Once registration had been completed, DICOM files were used with ORS Visual software (v1.5, Montreal), which allowed multiple display and post‐processing possibilities (3D VR; orthogonal/curved multiplanar reformation). Due to large size files, image processing was performed on PC with Windows 7 (64 bit) and 20 GB RAM. Three‐dimensional and multiplanar study of the facial nerve morphogenesis was possible with appropriate thresholding. The size, shape and positional changes of the facial nerve, geniculate ganglion, greater petrosal nerve and chorda tympanic relative to the external and middle ear were studied. Human embryonic facial nerve morphogenesis works are very rare because of the smallness of this anatomical structure. Our study using historic human embryos histological sections has shown the facial nerve growth. The precise knowledge of this morphogenesis will help to better understand congenital malformations.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The Infraorbital Foramen Location in the Perinatal Maxilla: Implications for Infraorbital Nerve Block
Authors: Ritz Brianna S , Nestor Nicholas S , Zdilla Matthew J
Published: June 2021

Bilateral infraorbital nerve block is performed in infants for the management of pain after cleft‐lip repair. The procedure involves the injection of anesthetic near the emergence of the infraorbital nerves in the face at the infraorbital foramen. In adults, infraorbital foramen location is oftentimes estimated via measurements from the lateral nasal aperture and the infraorbital margin; however, these measurements have not been described in the perinatal population. Therefore, this study assesses the distances from the lateral nasal aperture and the infraorbital margin to the infraorbital foramen in 66 fetal and infantile maxillae ranging from 5 months intrauterine development to 15 months postnatal development. The best‐fit linear model for IOF location is with regard to the distance from the lateral nasal aperture to the IOF. The model is given by the formula Distance = 0.1747 × Age (in developmental months) + 4.4433 and has an r2 value of 0.6145. Linear modeling with respect to the distance from the infraorbital margin to the IOF however was poor with regard to goodness‐of‐fit (r2 = 0.1636). The distance from the infraorbital foramen at 5 months intrauterine age was 2.9mm while, at one year post‐natal was 3.4mm (difference = 0.5mm) and therefore represents little change in maxillary growth in that dimension. On the other hand distance from the lateral aperture at 5 months development was 5.3 mm and at one year was 7.6mm (difference = 2.3mm). The results of this study suggest that the IOF location can be approximated, particularly with regard to the distance from the lateral aperture as a function of age. The results of the study will help to optimize infraorbital nerve block in young patients recovering from cleft‐lip surgery.

Support or Funding Information

WV Research Challenge Fund [HEPC.dsr.17.06]

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Quercetin hydrate promotes osteoblast differentiation and mineralization in vitro
Authors: Wyche Deanna , Walewska Klaudia , Belcher Joyce
Published: June 2021

Bone formation by osteoblast cells and osteoclastic bone resorption must be coupled throughout a lifetime to maintain homeostasis of the human skeleton. However as a normal part of the aging process, bone mass begins to decline due to enhanced osteoclasts activity. Osteoblast differentiation progresses through stages of cell proliferation, matrix deposition, and matrix mineralization. The identification of agents, which could regulate bone balance on the cellular level, would have potential therapeutic applications to regulate skeletal disorders. Quercetin hydrate (QH) is a non‐citrus flavonoid that has been previously been studied in its role as an antioxidant. Our study aimed to evaluate the direct effects of QH on osteoblast cell proliferation and differentiation in vitro. QH was added to the pre‐osteoblastic cell lines MC3T3‐E1 and MG‐63 at two concentrations. Cell proliferation and toxicity were assessed at 3 and 7 days of QH treatment via biochemical assays. Cell cultures were differentiated in vitro to 14 and 21 days with osteogenic media and stained to visualize matrix deposition and calcium/phosphate mineral maturation. Alkaline phosphatase was assessed at days 14 and 21 to assess osteoblast enzyme activity and function. Cultures treated with QH showed significant increase in cell proliferation and viability at both treatment doses. In response to QH treatment, there were no toxic effects observed. We observed more mineral matrix staining in QH‐treated cultures compared to untreated control. Von Kossa and Alizarin Red staining demonstrated enhanced osteoblast matrix deposition and mineralization associated with QH treatment. We observed an increase in Alkaline Phosphatase enzyme activity with QH treatment. Collectively, these findings suggest QH has the ability to promote osteoblast cell differentiation, matrix organization and maturation in vitro and could potentially promote skeletal health by acting in a bone anabolic fashion.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
New in vivo microRNA biotechnology reveals specific roles for the miR‐200 family in dental stem cell maintenance
Authors: Sweat Mason E , Sweat Yan Yan , Eliason Steven , Yu Wenjie , Cao Huojun , Hong Liu , Amendt Brad A
Published: June 2021

Extracellular signaling as well as transcription factor and epigenetic regulation are well described mechanisms controlling stem cell differentiation. The regulation of these processes by microRNAs (miRs) is recognized as an important biological process that can be coopted for future stem cell‐based therapies. Previously, our group has found the miR‐200 family is highly expressed in differentiated tooth cell types, but absent in dental epithelial stem cell (DESC) precursors. Inhibition of the miR‐200 family in mice using a novel Plasmid‐based miR Inhibitor System (PMIS) demonstrates that the miR‐200 family controls DESC maintenance and compartmentalization.

Objectives

To investigate the mechanisms by which the miR‐200 family promotes the differentiation of ameloblasts and other mature tooth cell types during tooth morphogenesis, and how inhibition of the family maintains stemness.

Methods

We generated mice expressing two different PMIS constructs. One inhibitor construct is specific for the seed sequence of miR‐200a, and ‐141, while the other inhibitor is specific for the seed sequence found in miR‐200b,‐200c and ‐429. Crossing these mice allows the functional inhibition of the entire miR‐200 family (double inhibitor mice).

Results

Double inhibitor mice are neonatal lethal and have poorly differentiated lower incisors. The teeth are smaller and lack markers of mature odontoblasts and ameloblasts. Several transcription factors marking the stem cell niche of the lower incisor, including Sox2, have expanded expression domains, and BrdU labeling of proliferating cells demonstrated that miR‐200 inhibition leads to increased epithelial cell proliferation outside of the stem cell niche. Rnf126, an E‐3 ubiquitin ligase that promotes P21 degradation and a direct target of miR‐200c, is expressed at higher levels in the proliferating cells leading to the loss of P21 protein. Although more epithelial cells are proliferating, the lower incisors are shorter due to an increase in a caspase independent, programmed cell death pathway.

Conclusions

The miR‐200 family is required for the appropriate differentiation of DESCs and the family functions by reducing the expression of transcription factors promoting stemness outside of the stem cell niche compartment of the lower incisor. Overall, craniofacial development is affected due to a lack of cell differentiation.

Support or Funding Information

University of Iowa College of Dentistry

NIH, NIDCR

T90 training grant 2T90DE023520‐06

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.