bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular Genetics of Human Sleep Behaviors
Authors: Fu Ying‐Hui , Ptacek Louis John
Published: June 2021

Sleep plays an essential role in our total well‐being and survival. Long‐term shift working schedule and frequent transmeridian travel have become inevitable for many in the current society. These factors lead to the out‐of‐sync condition of the sleep timing with the internal body clock, and thus significantly contribute to many health conditions that we are facing. Our group uses human genetics approach to identify genes/mutations that give rise to unusual sleep timing trait in human. Mouse models recapitulating human condition, coupled with in vitro molecular studies, offer new insight into the underlying mechanism. Because the fundamental role that sleep plays in our health, part of the pathways for regulating sleep are intertwined with pathways for regulating other functions. Our method also offers opportunities to investigate how sleep can impact other conditions including mood and pain regulations. These genes/mutations have provided unique opportunities to reveal the molecular mechanism of human sleep timing regulation and beyond. Previously, we have reported mutations on PER2, CRY2, PER3, and CK1delta that cause familial advanced sleep phase in human. Interestingly, the main components of molecular clock are mostly conserved from flies to mammals; however, the role of mammalian Timeless remains ambiguous. We recently identified a mutation on human TIMELESS gene for advance sleep phase trait. Behavioral and molecular characterizations of these mutations will be presented.

Support or Funding Information

This work was funded by NIH grants NS072360 and HL059596 to Y‐H.F. and L.J.P., NIH grant P30 DK063720, and by the William Bowes Neurogenetics 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.
Radiation Cystitis Modeling: a Comparative Study of Bladder Radiation‐Induced Fibrosis in Different Mouse Strains
Authors: Zwaans Bernadette MM , Wegner Kyle A , Bartolone Sarah N , Chancellor Michael B , Vezina Chad M , Lamb Laura E
Published: June 2021

Introduction and Objective

Radiation cystitis (RC) is a severe side effect from radiation therapy that can occur in pelvic cancer survivors (e.g. cervical and prostate cancer) and results in a fibrotic bladder. However not all patients treated with pelvic radiation will develop RC. Different mouse strains can be used for exploring genetic variability with respect to human biology. We aimed to assess the fibrotic response of the bladder to radiation in various genetic mouse strains to determine if genetic background variation may explain differences in risk for patients developing RC after radiation treatment.

Methods

Female C57BL/6, C3H and Balb/c mice were given a single dose of 40 Gy irradiation using the Small Animal Radiation Research Platform (SARRP) to deliver a gamma‐irradiation to the bladder while limiting toxicity to neighboring tissue. Weekly micturition measurements were performed. Mice were sacrificed one‐year post irradiation and bladders were processed for histology. Fibrosis was determined using Mason's trichrome and Picrosirius Red stains.

Results

All mice tolerated the radiation treatment well ‐ no changes in weight or behavior were noted. Micturition measurements were performed in C57BL/6 and Balb/c mice only, as C3H mice destroyed the filter paper. Increased number of voids, and decreased average and primary void size developed in both strains, though occurred earlier in the C57BL/6 strain. Hematuria developed in all strains. Mild to severe radiation‐induced fibrosis was visible in Balb/c and C57Bl/6 mice, with the highest degree of fibrosis in the latter. Fibermetrics analysis indicated higher number of collagen fibers in C57Bl/6 and Balb/c mice, but only Balb/c mice showed increased collagen fiber length and diameter. Ongoing studies will determine changes in specific collagen proteins using antibody specific fluorescent staining.

Conclusion

Using SARRP, we can create bladder dysfunction in mice modeling symptoms of chronic human RC including hematuria, fibrosis, and micturition frequency. Radiation‐induced fibrosis in the bladder is strain dependent, suggesting that there may be a genetic predisposition to radiation sensitivity of the bladder. These results are essential in determining the right preclinical model for future RC studies and understanding which cancer patients are at risk for RC.

Support or Funding Information

This project was in part supported by the U Can‐Cer Vive Foundation, NIDDK K01 career development award DK114334 (BMMZ), NIDDK specialized center cooperative agreement award U54DK104310, and NIEHS predoctoral research award F31ES028594 (KW)

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Self‐Antigen‐Specific T Cell Receptor Activation of CD4+ T Cells in the heart is required for Maladaptive Cardiac Remodeling in Pressure Overload Induced Heart Failure
Authors: Ngwenyama Njabulo , Carrillo‐Salinas Francisco , Aronovitz Mark , Alcaide Pilar
Published: June 2021

Cardiac pressure overload is associated with an adaptive immune response that drives maladaptive cardiac remodeling. CD4+ T cells are activated and expanded specifically in the heart‐draining mediastinal lymph nodes (mLNs) and infiltrate the left ventricle (LV) in response to transverse aortic constriction (TAC), a well‐established mouse model of heart failure (HF). In contrast, T cell deficient mice (Tcrα−/−) do not develop HF in response to TAC. Classic CD4+ T cell activation occurs upon cognate antigen recognition on antigen presenting cells, such as dendritic cells (DCs), through the T cell receptor (TCR). However, the specific mechanisms triggering T cell activation during the progression of HF remain unknown. We hypothesized that TCR‐dependent activation of CD4+ T cells by self‐antigens initiates and sustains maladaptive cardiac remodeling during the progression of TAC induced HF. We report a three‐fold increase in DC activation by flow cytometry in both the LV and mLNs in response to TAC, as compared to Sham surgery. Major histocompatibility complex class II deficient (MhcII−/−) mice, which lack the ability to present antigens to CD4+ T cells, did not develop cardiac dysfunction upon reconstitution with effector CD4+ T cells in the onset of TAC, despite developing some signs of adverse cardiac remodeling. A similar phenotype was observed in transgenic OT‐II mice, which undergo TCR activation only in response to exogenous ovalbumin (OVA), upon immunization with OVA during TAC. We next directly assessed whether CD4+ T cells respond to endogenous antigens in the heart using Nur77GFP mice, which transiently express GFP upon TCR stimulation. Strikingly, GFPhiCD4+ T cells were present in the LV, and increased in number as maladaptive remodeling and cardiac dysfunction progressed over time. TCR clonal analysis by next generation sequencing of TCR‐activated GFP+CD4+ T cells sorted from the LV demonstrated a limited repertoire of TCR clones relative to the periphery, suggesting that an antigen‐restricted response occurs in the LV in response to pressure overload. Taken together, our results indicate that CD4+ T cell recognition of self‐antigens through the TCR is required for maladaptive cardiac remodeling and cardiac dysfunction. Ongoing studies are focused on the identification of specific self‐antigens generating this clonal CD4+ T cell response in the heart.

Support or Funding Information

NIH R01 HL123658‐01, T32 AI007077‐34, F31 HL140883‐01A1, AHA 19PRE34020084

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Defining the Role of the Sympathetic Nervous System in Leukocyte Migration In Vivo
Authors: Weber Jasmin
Published: June 2021

The nervous system and the immune system are intricately linked, with neurotransmitters controlling many aspects of leukocyte biology. Recently, we have shown that leukocyte migration patterns from murine blood to tissues fluctuate over 24h, exhibiting peak numbers in blood during the day and a trough in the night. How neural signaling pathways are involved in these circadian oscillations remains to be elucidated. Here, we investigated the effect of sympathetic nervous system manipulations on the circadian migration behavior of leukocyte subsets in vivo. Stimulation of specific β‐adrenoreceptors drastically influenced blood leukocyte numbers in a subset‐specific manner. Using systematic adoptive transfer assays of leukocytes combined with flow cytometric analyses we could attribute this to an increased capacity of blood leukocyte subsets to emigrate into organs. We furthermore investigated this migratory homing behavior of leukocyte subsets in real time using intravital microscopy of different tissues and focusing on the effect on different steps of the leukocyte adhesion cascade. Together, our investigations provide mechanistic insights into the leukocyte‐subset specific migration behavior after sympathetic stimulation.

Support or Funding Information

We are very thankful for our funding agencies: European Research Council (ERC), Deutsche Forschungsgemeinschaft (DFG), Integrated Research Training Group 914 / Sonderforschungsbereich 914 of the Ludwig‐Maximilians‐Universitaet Muenchen, International Max Planck Research School for Life Sciences

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Bacteria degrade the specialized basal lamina attaches the gingiva to teeth
Authors: Fouillen Aurélien , Grenier Daniel , Barbeau Jean , Baron Christian , Moffatt Pierre , Nanci Antonio
Published: June 2021

Introduction

The junctional epithelium is a specialized portion of the gingiva that seals off the tooth supporting tissues from the oral environment. This relationship is achieved via a specialized basal lamina (sBL). Three unique proteins ‐AMTN, ODAM and SCPPPQ1‐ together with Laminin‐332 structure the supramolecular organization of this sBL and determine its adhesive capacity. Despite its critical and strategic importance and continued exposure to bacteria, little is known on the susceptibility of the sBL to bacterial activity.

Methods

The objective of this study was to evaluate the impact of various oral bacteria on the sBL using molecular biology and complementary imaging approaches.

Results

SDS‐PAGE and western blot analysis of degradation assays with trypsin like proteases as well as incubation with Porphyromonas gingivalis (P. gingivalis) revealed that all sBL constituents, except SCPPPQ1, were rapidly degraded. Mass spectrometry also confirmed that SCPPPQ1 was not degraded. We have previously shown that mixing the recombinant sBL proteins results in the formation a supramolecular network that mimicked the sBL. To observe the impact of oral bacteria on this ‘reconstituted’ sBL network, we exposed it to P. gingivalis and observed the results by atomic force microscopy and scanning electron microscopy (SEM). The bacteria destroyed the supramolecular network in their vicinity creating a peripheral space around them. SEM imaging further showed the presence of fine filamentous residual matrix on the exposed HOPG surface, suggesting the incomplete degradation of some component and a layered destruction of the network. Finally, to determine the effect of P. gingivalis on the native sBL itself, we exposed it ex‐vivo to bacteria. After two hours of exposure, SEM analysis revealed the presence of a shallow depression around the bacteria and after 6 h, the affected peri‐bacterial area became deeper and cavitated , suggesting active degradation of the native sBL as well.

Conclusion

We show here for the first time that enzymes and P. gingivalis, one of the major bacteria implicated in chronic periodontitis, can attack the individual components of the sBL as well as alter the supramolecular organization of this critical adhesive extracellular matrix. Because destruction of the sBL would open the door to bacterial infiltration around teeth. These results highlight the importance of the sBL in PD and open the door for new treatment paradigms.

Support or Funding Information

This work was supported by CIHR, Canada Research Chair, RSBO, and Shriners Hospital.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Methicillin‐resistant Staphylococcus aureus causes sustained collecting lymphatic vessel dysfunction
Authors: Jones Dennis , Padera Timothy
Published: June 2021

Methicillin‐resistant Staphylococcus aureus (MRSA) is a major cause of morbidity and mortality worldwide and is a frequent cause of skin and soft tissue infections (SSTIs). Lymphedema—fluid accumulation in tissue caused by impaired lymphatic vessel function—is a strong risk factor for SSTIs. SSTIs also frequently recur in patients and sometimes lead to acquired lymphedema. However, the mechanism of how SSTIs can be both the consequence and the cause of lymphatic vessel dysfunction is not known.

Here, we used the endemic USA300 strain of community‐associated MRSA (CA‐MRSA), a common cause of serious bacterial infections in the United States, to establish a model of localized MRSA infection. We used intravital imaging to determine whether MRSA infections inhibit lymphatic vessel contractility and lymph flow. In vivo studies used C57BL/6, iNOS−/− C57BL/6, MyD88−/− C57BL/6, LysMgfp/gfp C57BL/6, and alpha SMAP‐DsRed/C57BL/6 mice to assess the effects of host‐derived molecules to lymphatic contractility after infection. Alpha SMA‐DsRed transgenic mice were visualized and immunofluorescence staining of alpha SMA+ lymphatic muscle cells (LMCs) of WT mice was performed to analyze LMC coverage of lymphatic vessels on multiple days after infection. To define the molecular mechanisms related to this response, we performed cell viability and cell lysis assays and measured killing of LMCs by MRSA‐conditioned supernatant from wild type and mutant MRSA strain(s). Similar experiments were performed using recombinant MRSA toxins.

Intravital imaging in mice revealed an acute reduction in both lymphatic vessel contractility and lymph flow after localized MRSA infection. Moreover, chronic lymphatic impairment is observed long after MRSA is cleared and inflammation is resolved. Associated with decreased collecting lymphatic vessel function was the loss and disorganization of lymphatic muscle cells (LMCs), which are critical for lymphatic contraction. In vitro, incubation with MRSA‐conditioned supernatant led to LMC death. Proteomic analysis identified several accessory gene regulator (agr)–controlled MRSA exotoxins that contribute to LMC death. Infection with agr mutant MRSA resulted in sustained lymphatic function compared to animals infected with wild‐type MRSA.

Our findings suggest that agr is a promising target to preserve lymphatic vessel function and promote immunity during SSTIs.

Support or Funding Information

This work was supported by the National Institutes of Health under award numbers R21AI097745, DP2OD008780, R01CA214913, and R01HL128168 to Timothy Padera. Dennis Jones was supported in part by the UNCF‐Merck Science Initiative Postdoctoral Fellowship, Burroughs Wellcome Fund Postdoctoral Enrichment Program Award, and NIH NCI F32CA183465.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Clinical Cases Review: Hand Innervation and Function using a Hand Model
Authors: Elmansoury Amr , Pessina Monica
Published: June 2021

Objective

To improve the understanding of the anatomy of the hand muscles and its nerve supply.

To understand the different types of nerve injuries of the hand

Application of the theoretical knowledge on a hand model

Material and Methods

Twenty‐eight first‐year medical students had a 55 minutes educational session that included

‐ A pretest

‐ Ten minutes focused power point presentation.

‐ An interactive simulation of hand deformities using a color labelled hand model

‐ Post test

‐ Feedback questions

Results

Twenty‐eight students participated in the session. Scores were measured on a scale from 0–8. The mean pretest score was (3.4 +/− 1.7) while the posttest score was (5.07 +/− 2.25). The posttest scores were higher than the pretest scores, and that was statistically significant using a paired t‐test of P = 0.002

Conclusion

Using a short presentation and an interactive simulation of the hand function impairment and anatomy was effective in teaching various nerve injuries of the hand.

Support or Funding Information

No funding was received

Pre and Post Test Score Chart

image

Pre and Post Test Score Chart

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 review of U.S. medical schools promotion standards for excellence in education
Authors: Hoffman Leslie , Lufler Rebecca , Brown Kirsten , DeVeau Kathryn , DeVaul Nicole , Fatica Lawrence , Mussell Jason , Byram Jessica , Dunham Stacey , Wilson Adam
Published: June 2021

Purpose

The number of medical science educators, who commit the majority of their time to education‐related duties, is ostensibly on the rise. Given the growing role separation between teaching and biomedical research faculty in medical education, it is timely to examine how institutions communicate their promotion criteria related to educational excellence and scholarship. This study investigates the extent to which medical schools' promotion criteria align with published standards for documenting educational activities.

Methods

Promotion guidelines from U.S. allopathic and osteopathic medical schools were collected mostly from institutional websites and were systematically analyzed according to a predefined data extraction rubric adapted from previously published standards for documenting educational activities. After 10 researchers each independently reviewed and extracted data from 1/5 of all guidelines, researchers compared their findings in pairs and reached consensus on all identified discrepancies prior to final data submission. Descriptive statistics were used to determine the frequency of cited promotion criteria.

Results

Promotion‐related documents were retrieved for 110 (59%) of the 185 allopathic and osteopathic U.S. medical schools. Fifteen percent of schools were cited as lacking explicit direction for education focused faculty to attain academic advancement. Across seven education‐related domains, ‘educational measurement and evaluation’ was the least represented with only 37% of schools referencing this domain in their guidelines. The other domains including teaching, curriculum/program development, mentoring/advising, educational leadership/administration, research/scholarship, and service were referenced by at least 59% of schools. Overall, only 20% of schools were judged to have above average or very comprehensive criteria for excellence in education.

Conclusions

While most medical schools acknowledge education within their promotion criteria, only three fully embraced the educational excellence standards recommended in the literature. These data should be concerning to current teaching faculty who may be evaluated for promotion based on vague and/or incomplete promotion criteria. With greater awareness of how educational excellence is currently documented and how promotion criteria can be improved, more schools may be compelled to embrace change and to follow recommended best practices.

Support or Funding Information

None

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Class Attendance and Student Performance in Medical School Anatomy Education
Authors: Fang Xiaoqian , Olvera Nina , Madrigal Karina
Published: June 2021

Student academic performance in medical school is positively correlated to their success during their graduate medical education, as well as their career in the short term. Improving student academic performance is an essential part of educational research. A significant decline of classroom attendance in medical school has been noticed with the trend of non‐mandatory classes and the availability of lecture videos. In this study, we used the data collected in anatomy education to investigate the relationship between students' classroom attendance and academic performance. Since the attendance for lectures is optional, the attendance data was collected in the mandatory laboratory sessions by circulating an attendance sign‐in sheet or through an electronic application (TopHat). Students were divided into three groups, 1) students who attended the session less than the mandatory requirement, 2) students who attended all sessions, and 3) students who attended the session more than the mandatory requirement. This study included one‐hundred students, covering eight different modules, including both year 1 and year 2 medical students. We find that (a) group 3 always performed better than group 2 in written exams; (b) group 3 always performed better than group 2 in lab practical exams; (c) group 1 performed better in the beginning of the medical school study even though students attended mandatory lab sessions less frequently than their peers; (d) from the third module onwards, group 1 students started performing poorer than the other two groups and remained the same till the end of the second year. The main conclusion reached by this study was that classroom attendance positively correlates to student academic performance. This study highlights that student attendance can be used as a predictor to estimate student academic performance and can help educators identify and facilitate interventions for at‐risk students.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Examining Color Vision Deficient Health Professional Student Performance and Attitudes in Histology
Authors: Weinfeld David , Granadier Robert , Attardi Stefanie M.
Published: June 2021

All United States medical and dental schools incorporate histology into their curricula. Histology is the microscopic study of tissues and often involves specific colored staining. Histology's role in health professional education and practice underscores the need for students to understand and apply histological concepts. It has been proposed that color vision deficient (CVD) students may struggle learning histology and it is estimated that 8% of men and 0.5% of women in the general population are CVD. No studies have quantifiably assessed students' color vision and performance in histology. This research objectively assesses health professional students' color vision and performance in histology, as well as obtains subjective information about student attitudes and experiences about histology. The objectives of this study are to compare normal visioned to CVD students in terms of performance and time on task on an image‐based histology assessment and to compare their histology study experiences. It is hypothesized that compared to the normal visioned students, CVD students are expected to spend significantly more time on the image‐based questions and score significantly lower. It is also expected that CVD students will be unaware of their disadvantage in histology; thus, are likely to seek help from professors at similar rates as their normal visioned peers. To date, first‐year and second year medical students at Oakland University William Beaumont School of Medicine and first year students at University of Michigan (U of M) Medical School and U of M School of Dentistry have been invited to participate in the study. Participants (n= 72) completed an online Qualtrics survey consisting of demographic questions, a validated colorblindness examination, 52 multiple choice image‐based histology questions and a post‐survey on student attitudes and study experiences in histology. Analyses will compare CVD students and normal visioned students on time spent, correct answers (independent T tests), and attitudes and experiences in histology (Mann‐Whitney U). Recruitment is ongoing to increase responses from the small population of CVD students. The results are expected to quantify that CVD medical students are both unaware of their disadvantages in histology and receive lower scores on histology assessments relative to their normal visioned peers. This information would show histology and health professional educators the need to specifically identify and accommodate CVD students, who otherwise may not know to seek help with their learning.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of First Year Medical Students' Performance on First and Second Order Questions on Gross Anatomy Practical Examinations
Authors: Ferriby Andrew Charles , Williams Sutton , Sinning Allan
Published: June 2021

The goal of this study was to determine how first year medical (M1) students perform on different question types, first and second order, during gross anatomy practical examinations. In this study first order are simple “identify the tagged structure” questions. Second order questions require students to identify a structure, and apply their knowledge of it to answer correctly. An example of a second order question is “What spinal nerve levels does the tagged nerve originate from?” To conduct this investigation, laboratory practical examination questions (N=114) were coded as either first order (N=96) or second order (N= 18). Responses from the 2018 M1 class (N=167) were entered (1= correct, 0= incorrect) for analysis. The responses were added up and averaged for a first order, second order and overall examination grade. The overall average for the first half of the course was 80.66%. Averages for first and second order questions were 81.95% and 73.75% respectively. This resulted in an overall difference of 8.2% between the two different question types. A further break down shows a difference of 8.42% on block 1 and 6.45% on block 2. Data collection for the second half of the course is ongoing and will be added as it is obtained. These preliminary findings suggest that M1 students have a lower performance rate on second order questions. This may be an indicator that M1 students are not learning the material in a way that allows them to apply content effectively to answer second order questions. This apparent difficulty in students' ability to answer these questions is important for faculty to consider both in assessing instructional design and students' knowledge. Further investigation may yield more conclusive answers as to what may cause this disparity and how to address it.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Comparison of the “Histo!” app with traditional histology study guides
Authors: Moore Kathryn , Colombo John , Ash Rick , Scott Sheryl
Published: June 2021

Objectives

“Histo!”, an iPad application, was developed by histology faculty at the University of Utah (UU) in response to student requests for a histology learning tool. The intent was to provide a self‐directed learning app for medical and dental students using high‐quality histological images from the UU histology slide collection. Histo! was designed to complement other teaching methods and facilitate initial learning, review, and self‐testing of course material. Content includes structural and functional questions, incorporating learning, quiz, and test modes. Many students use Histo! as their main study tool. Although there are many basic science educational apps available and several histology apps, few have been validated and tested for dental students.

Methods

In this pilot study, we assessed the educational value of Histo!, specifically whether the app functions better as a tool for learning, review, or both. 48 first‐year dental students took a pre‐test assessing their baseline histological knowledge of different body systems. Students were randomly sorted into groups and for the week preceding each lab each group of students prepared by using either the Histo! app or provided lab study guides, containing the same content. Before each lab, students retook the topic test. Group scores were compared using Student's T‐test. Groups were assessed on four different topics, with preparation of two topics using Histo! and the other two using study guides. At the end of the study, a qualitative survey was given to students to assess their usage and learning preferences regarding the Histo! App vs. traditional study guides.

Results

Comparison of group scores on the post‐tests showed a 2‐fold improvement in scores, irrespective of study method. There were no statistically significant differences in group performance (p= 0.24), though the Histo! group scores trended slightly higher. Students were surveyed on their usage of Histo! throughout the course, comparing the two modes for initial learning and as an exam review tool. In addition, students gave feedback on their sense of preparation, satisfaction and engagement with the app. Final results from the qualitative student experience survey are still pending.

Conclusions

Preliminary results suggest that Histo! is as effective as study guides, at least for initial learning. Students found the app to be engaging and less time consuming than traditional study methods, and preferred spending study time on the app as opposed to using the traditional study guides.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Learning through the Eyes of the Beholder: Using Eye Tracking to Understand How Novices Learn Neuroanatomy
Authors: Leclair Rebecca , Nguyen Angela , Cheung Beata , Brewer‐Deluce Danielle , Heisz Jennifer , Lyons Jim , Wainman Bruce
Published: June 2021

Introduction

Despite student reports of high difficulty and neurophobia, there remains little examination in the literature on what makes neuroanatomy challenging. A preliminary study found that increased grey‐to‐white matter stain contrast marginally improves learning in students with low working memory capacities (WMC) but not those with high WMC. Why this augmentation is beneficial to some but not all students is unknown. Further, evidence suggests competency influences the way images are viewed and interpreted, which may account for some this previously observed difference.

Aim

This study will use eye tracking to better understand how novice students view and learn neuroanatomy. Specifically, we aim to: 1) assess differences in viewing patterns between high and low contrast brain slices for students with high and low WMC, and 2) to then compare viewing patterns between novices and experts to uncover competency‐related changes as they relate to contrast.

Methods

Undergraduate students with no previous anatomical education (n=120) were recruited to complete an eye‐tracking session containing two learning and two testing periods. Each learning period consisted of 4 brain slices (either coronal or transverse planes and high or low contrast) labeled with 12 neuroanatomical structures. The students were given 5 minutes to learn the structures while their viewing patterns were tracked. In the testing periods, students were prompted to click on named structures on a low‐contrast brain slice. Eye tracking data was recorded along with test accuracy. This learning/testing protocol was repeated twice such that the brain image sectioning plane (coronal vs transverse) and the stain contrast (high vs low) order of exposure were counterbalanced. Finally, participants completed the Automated Operation Span Task (OSPAN) to quantify their WMC.

Results

Data collection is ongoing. Preliminary results suggest differences in fixation durations, frequency of fixations on irrelevant areas, and time to first fixate on relevant structures between novices and experts. Full data will be subsequently analyzed and presented.

Discussion/Conclusion

By better understanding how students learn neuroanatomy, we will be able to gain insight into factors that contribute to the difficulty of the subject and support research on better methods for teaching neuroanatomy.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Photogrammetry or 3D Surface Scanning – Which tool works best for anatomical specimens?
Authors: Dixit Ishan , Piemontesi Joshua , Kennedy Samantha , Kennedy Bruce , Krebs Claudia
Published: June 2021

High fidelity 3D representations of anatomy are becoming increasingly popular as educators capture anatomical specimens for use in virtual and augmented reality applications or for providing a 3D representation that can be freely manipulated in web applications.

The question then arises which tool best suits the task at hand ‐ both 3D scanning and photogrammetry are options.

We have compared the use case for these technologies in medical education ‐ our aim was to create 3D models of anatomical specimens with high quality and resolution. Various qualitative and quantitative criteria were applied to determine and compare the performance fidelity and results of 3D scanning with an Artec Space Spider versus photogrammetry using a standard DSL camera and Agisoft PhotoScan (PS) Standard Edition. We compared the basic principles of setup and use of both technologies – while photogrammetry requires accurate lighting and some rigging for the camera to provide consistent results, the 3D surface scanner is handheld and since it acts as its own light source the scanning results are independent of environmental factors. We found that the workflow and technique for best use of these technologies varies and the investment in these techniques depends on the needs of the user. We established protocols and setting for both techniques to standardize and streamline the data acquisition process. Each technology has advantages in some metrics and disadvantages in others. While photogrammetry provided the better surface textures, 3D surface scanning provided more accurate 3D geometries in particular in those specimens that were more complex or had highly reflective surfaces. We also found that 3D surface scanning was able to more accurately capture the geometries of softer specimens that may distort as they are handled.

Overall, both technologies yield excellent results for the use in anatomical sciences education. The decision about which technology to invest in depends on the characteristics of the specimens that need to be scanned as well as the environmental parameters.

Support or Funding Information

Supported by the Strategic Investment Fund of the UBC Faculty of Medicine

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Determining the moving signals from the brain motor area for imaging applications
Authors: Ramirez Jennifer Cristina , Manian Vidya
Published: June 2021

Brain‐computer interfaces (BCI) allows a noninvasive connection between the human brain and a computer system. This communication capability is useful for disabled people with impairments such as spinal cord injuries and neuromuscular disorder. In order for the BCI to acquire information, electrodes are used on certain localizations of the scalp to read the electrical brain activity. However, some challenges with BCI includes understanding key areas of the brain. Here, using Python and Psychtoolbox, we investigate the motor area of the brain to find how disabled people respond to moving targets compared to healthy subjects. The outcomes of this project will be an analysis of brain signals elicited in response to moving targets. The response of the motor area to moving signals for imagining limb movements will be investigated in healthy and disabled subjects. This may lead to better motor imagery signals for control of BCI.

Support or Funding Information

Financial support funded from the NIH RISE‐E‐BASE program grant number 1R25GM127191‐01

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Altered striatal dopaminergic system in heterozygous Disc1 mutant mice
Authors: Baskaran Rathinasamy , Lee Li‐Jen
Published: June 2021

Disrupted‐in‐Schizophrenia‐1 (DISC1) is a susceptibility gene for several psychiatric illnesses. To study the pathogenesis of these disorders, various Disc1 mutant models were established. We had generated Disc1 mutant mice by introducing the 129S6/SvEv 25‐bp deletion Disc1 variants into C57BL/6J strain. In this study, a battery of behavioral tests was conducted to evaluate the behavioral phenotypes and cognitive function in the heterozygous Disc1 mutant (Het) mice. In open field test, novel object recognition test, Y‐maze test, and prepulse inhibition test, no significant differences were observed between wildtype (WT) and Het mice. However, after receiving an intraperitoneal injection of amphetamine (5 mg/kg), greater locomotor activity was noticed in Het mice comparing with WT. Meanwhile, higher amphetamine‐induced c‐fos expression and dopamine level were found in the striatum of Het mice than those in WT mice, suggesting an altered dopaminergic system in the striatum of Het mice. We then quantified the expression of proteins involved in dopaminergic function and DISC1 signaling in the striatum. Tyrosine hydroxylase (TH) level was not changed; dopamine transporter (DAT) and D2 dopamine receptor (D2DR) were increased while D1 dopamine receptor (D1DR) was decreased in Het mice. The expression of glycogen synthase kinase 3 (GSK3α/β) was down regulated in Het mice while some DISC1 interacting proteins such as phosphodiesterase 4B (PDE4B), cAMP response element binding protein (CREB) were not altered. Morphologically, the dendritic complexity and spine density of striatal median spiny neurons were reduced in Het mice. Our results indicated that Disc1 deficit in one allele does not alter the basic behavior and cognitive performance in mice. However, mice lacking one WT Disc1 allele are vulnerable to psychostimulant such as amphetamine, which might be attributed by altered structure and function of striatal dopaminergic system. Here, we demonstrated striatal phenotypes in heterozygous Disc1 mutant mice which could be a promising animal model of DISC1 haploinsufficiency.

Support or Funding Information

This work was supported by the Ministry of Science and Technology of the Republic of China; Grant number: 102‐2628‐B‐002‐014‐MY3.

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Impact of Adolescent Intermittent Ethanol on Noradrenergic Development
Authors: Coker Caitlin Rae , Aguilar Elizabeth , Silberman Yuval
Published: June 2021

In clinical populations, adolescent alcohol drinking has been shown to increase stress sensitivity and the likelihood of developing alcohol use disorders (AUDs) in adulthood. These findings suggest pathways mediating stress‐relating behaviors are impacted by developmental exposure to alcohol, contributing to a greater risk for alcoholism and potentially stress‐induced relapse following treatment. Animal models indicate tyrosine hydroxylase (TH) neurons in the A2 region of the nucleus of the tractus solitarius (NTS) contribute to stress‐induced relapse through enhanced norepinephrine signaling, subsequently increasing neuronal activity in the bed nucleus of the stria terminalus (BNST). Previous research suggests NTS TH expression undergoes developmental changes during adolescence that may make this neuronal population susceptible to modulation by alcohol during this age. We hypothesized adolescent intermittent ethanol (AIE) exposure in mice, a standard preclinical model of adolescent binge drinking, will impact NTS TH neuron development, ultimately altering the NTS‐BNST circuitry resulting in a higher risk of AUDs in adulthood. NTS TH mRNA expression in male and female C57Bl6/J mice exposed to AIE (postnatal day [PND] 28–42) or used as air controls was quantified via qRT‐PCR at different time points (PND29, PND36, PND43, PND49, PND70). Immunohistochemistry methods were implemented to determine the impact of AIE on NTS TH neuronal activity following an ethanol challenge in adulthood (PND70). Results showed a significant increase in TH mRNA in males one day after beginning AIE (PND29) relative to air controls, while females exhibited the greatest TH expression after their first week of exposure to ethanol (PND36). In both sexes, heightened TH mRNA expression occurred earlier than expected during normal development. Surprisingly, compared to adult ethanol naïve mice, AIE decreased the ability of ethanol at doses of 2g/kg and 4g/kg to alter NTS TH neuron cFos expression at PND70 compared to control. Together, these findings suggest an enhancement of NTS TH activity early in development and reduction of ethanol sensitivity in adulthood. Studies are in progress to further delineate the mechanisms by which AIE reduces adult ethanol sensitivity and how this may alter stress‐related behaviors.

Support or Funding Information

AA22937

AA017823

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Human Apolipoprotein E Isoforms Influence Neurite Outgrowth and Regeneration In Vitro and In Vivo
Authors: Maggard Rachel Susanne Jolly , Calulot Christopher M. , Hager Lydia E. , Ritter Kyle J. , Turba Brittany N. , Hoffman Jared D. , Lin Ai‐Ling , Johnson Lance A. , Alilain Warren J.
Published: June 2021

Translating spinal cord injury (SCI) therapies which promote axonal regeneration from preclinical animal models into the human population is challenging. One potential obstacle is that human genetic predispositions may limit the efficacy of such experimental treatments. The clinically relevant ApoE4 (E4) allele, present in about 14% of the human population, corresponds to an increased incidence of Alzheimer's disease—however, its role in recovery from SCI is poorly understood despite suggestive data implicating its involvement. Indeed, two clinical studies found that SCI individuals with the E4 allele had less motor recovery than individuals without the allele despite longer time in rehabilitation. ApoE4 may mediate this diminished recovery by limiting regeneration and sprouting. Robust regeneration is energy intensive and requires efficient mitochondria, and studies have shown that ApoE4 impairs mitochondrial function. Given these mitochondrial deficits, we hypothesize that ApoE4 can impair regeneration and sprouting. To test this hypothesis, we investigated the impact of ApoE4 on sprouting and neurite outgrowth. In our experiments, we cultured dorsal root ganglia neurons from mice expressing human ApoE isoforms—ApoE2 (E2), ApoE3 (E3), or ApoE4—under the control of the endogenous mouse ApoE promoter. We then analyzed differences in 1) neurite complexity and 2) robustness of outgrowth between genotypes. In two of three independent experiments, E4 neurons had decreased neurite outgrowth and decreased neurite branching compared to E2 and E3 neurons. Data from the Spot Assay, an in vitro model of the glial scar and CNS regeneration, also suggest that chondroitin sulfate proteoglycans may inhibit regeneration in E4 neurons to an even greater extent than in E3 neurons. In addition, with preliminary in vivo data, we are beginning to characterize serotonergic sprouting after lateral C2 hemisection in mice of each ApoE genotype. Since outgrowth, sprouting, and regeneration all partially mediate recovery after CNS injury, impairments in these processes can adversely affect recovery. These foundational studies address not only the possible genetic influence of ApoE4 on recovery from CNS injury, but also a critical gap in knowledge—whether there is a genetic contribution underlying responses to treatment in SCI individuals.

Support or Funding Information

University of Kentucky Startup Funds

This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Lipopolysaccharide Differentially Alters Oligodendrocyte Precursor Cell Proliferation
Authors: McKinley Raechel Elaine , Ghaderzadeh Sadaf , Csoka Antonei
Published: June 2021

Multiple Sclerosis (MS) is a debilitating autoimmune disease that impacts 2.3 million people worldwide. In the pathogenesis of the disease, specialized nervous cells called Oligodendrocytes (OLs) are targeted and damaged. When OLs are damaged by the immune system, myelin is destroyed resulting in neuronal cell death. In the adult brain, pools of progenitor cells called Oligodendrocyte Precursor Cells (OPCs) are proliferated, migrated and differentiated to replace and repair mature OLs in the process called remyelination. However, in MS this process does not occur properly. Many factors such as inflammation contribute to the lack of proper remyelination. Moreover, in MS patients, it has been shown that there are increases in Toll‐Like receptors (TLRs). To determine if TLR2 and TLR4 activation plays a role in the proliferation step of remyelination, we treated Induced Pluripotent Stem cell derived OPCs with LPS and hypothesized that LPS treatments will increase the proliferation of OPCs through TLR4 and TLR2 activation. To analyze cell proliferation, a methylthiazol tetrazolium (MTT) assay and immunocytochemistry staining was conducted. Altogether, we found that LPS altered OPC cell proliferation in a dose dependent manner.

Support or Funding Information

National Institute of Health (NIH) R25 Resource Grant. (1 R25 AG047843‐01)

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