bluuebunny/crossref_metadata_2025 huggingface dataset:
Different Duration of Diet Transitions from a High‐fat Diet to a Normal‐fat Diet before Pregnancy Deferentially Affect the Phenotypes of Offspring Obesity with a Sex‐dependent Manner
Authors: Fu Qiang , Zhou Yi , Xu Huiting , Olson Patrick , Xie Linglin
Published: November 2023

It is now well established that in utero and in early life exposure to under‐nutrition or over‐nutrition can disrupt normal growth and development and thus results in a higher risk for offspring obesity in adult life. Therefore, important information/evidence is urgently needed regarding if and how a pre‐pregnancy healthy diet strategy would be beneficial to obese women or women who have been on long‐term unhealthy western‐style diet on pregnancy outcomes. Unfortunately, no evidence‐based strategy regarding the optimal duration of the diet intervention for the women at childbearing age is currently available. We used a murine model to study whether switching from a high‐fat diet to a normal‐fat diet (H1N group) 1 week, 5 weeks (H5N group) or 9 weeks (H9N group) before pregnancy could lead to in utero reprogramming of female offspring obesity; comparator groups were offspring given a consistent maternal high‐fat diet (HF group) or normal‐fat diet (NF group) until weaning. Another group of offspring (the REF group), which were fed the control diet and were born to breeding pairs given the control diet, served as the reference group to show the normal ranges for a given characteristic. After weaning, all offspring were given the high‐fat diet for either 9 or 12 weeks before killed humanely. The female offspring of H1N dam were obese and the female offspring of HF and H5N dams were overweight, while the NF and H9N female offspring slowly gained body weight during experimental period and were only lightly but significantly higher than REF female offspring at week 12. However, only male offspring from H1N group, but not other groups, were obese; and the HF male offspring slowly gained body weight and was heavier than H5N, H9N NF or REF at week 12. There were significantly enlarged subcutaneous fat and gonadal fat pads in H1N male offspring versus male from any other dams. For female, only H1N offspring, but not other groups, enlarged gonadal fat pads, but not subcutaneous fat pad. We performed IPGTTs of the offspring at the end of week 9 and week 12 of the postweaning HF diet. At both time points, female HF, H1N and H5N, but not NF and H9N offspring were glucose intolerant. However for male, only H1N and H5N, but not NF, HF or H9N offspring were glucose intolerant. In addition, the glucose level of H5N male offspring was unable to return to basal level, suggesting a high risk for diabetes. In summary, our data suggested that a short term transition (H1N and H5N) were not beneficial to reduce offspring obesity, however a longer tern transition (H9N) reduced the risk for offspring obesity and the phenotypes of obesity were gender dependent.

Support or Funding Information

This study is supported by grants from the National Institutes of Health (NIH‐1R15HL117238 to LX and the American Heart Association (Scientist Development Grant13SDG14650009 to LX).
Regulation And Function Of Fatty Acid Desaturases
Authors: Ntambi James , Burhans Maggie S. , Guo Chang‐An , Bond Laura M.
Published: November 2023

Stearoyl‐CoA desaturase is a central lipogenic enzyme catalyzing the synthesis of monounsaturated fatty acids mainly oleate (C18:1) and palmitoleate (C16:1) which are the major monounsaturated fatty acids of membrane phospholipids, triglycerides, wax esters and cholesterol esters. Several SCD gene isoforms (SCD1‐4) exist in the mouse. While SCD1, 2 and 4 desaturate both 16:0 and 18:0 to 16:1n7 and 18:1n9 respectively, the SCD3 isoform has a substrate specificity and prefers desaturation of 16:0 to 16:1n7. 16:1n7 has been labeled as a lipokine that represses hepatic de novo lipogenesis and improves insulin signaling in muscle. We found that mice with a targeted disruption of the SCD3 isoform (SCD3−/−) exhibit dramatic brown adipose tissue (BAT) expansion and whitening with paradoxical increase in monounsaturated fatty acids and triglyceride accumulation. The alteration in fatty acid composition and lipid metanbolism causes reduced abundance of electron transport chain components and near ablation of UCP1 in BAT. Interestingly, this decreased protein abundance is not driven by repressed gene transcription and likely stems from protein instability and degradation. Consequently, the SCD3−/− mice have impaired cold tolerance and cannot maintain core body temperature during a 4ºC cold challenge past 6 hours. Meanwhile, the subcutaneous WAT of the KO mice shows increased expression of genes associated with fatty acid oxidation and thermogenesis. These observations suggest that SCD3 expression plays distinct roles in BAT and subcuteneous fat depots and help to elucidate how changes in monounsaturated fatty acids modulate metabolic activities in adipocytes.

Support or Funding Information

Supported by NIH grants RO1DK62388 and T32 GM07215.
Discovery of A Novel Class of Naturally‐Occurring Lipids with Anti‐Diabetic and Anti‐inflammatory Effects
Authors: Kahn Barbara , Yore MM , Syed I , Moraes‐Vieira PM , Zhang T , Herman MA , Homan E , Lee J , Chen S , Peroni OD , Hammarstedt A , Patel R , McGraw TE , Smith U , Saghatelian A
Published: November 2023

Increased adipose tissue (AT) lipogenesis is associated with enhanced insulin sensitivity. Mice overexpressing Glut4 in AT (AG4OX) have elevated AT lipogenesis and increased glucose tolerance in spite of obesity and elevated circulating fatty acids. To determine if the lipid profile contributes to improved glucose homeostasis in AG4OX, we performed lipidomic analysis of AT. This revealed a 16‐18‐fold increase in a novel class of lipids in AG4OX AT vs wildtype mice. Using a targeted Mass Spec approach, we identified 16 novel lipid family members with multiple isomers based on structural variations. These lipids are branched fatty acid esters of hydroxy fatty acids or FAHFAs. We studied the biologic effects of the isomers of palmitic acid hydroxy stearic acid or PAHSAs. These are present at highest levels in brown adipose tissue followed by subcutaneous and perigonadal white AT with lower levels in many other tissues. PAHSA levels are acutely regulated by fasting. Most isomers are reduced 50–65% in serum and AT of insulin‐resistant vs insulin‐sensitive people. Nearly all isomers in humans correlate remarkably strongly with insulin sensitivity determined by euglycemic clamp. PAHSAs are also reduced in subcutaneous white AT in mice fed a High Fat Diet. Oral administration of PAHSAs lowers ambient glycemia and enhances glucose tolerance and insulin sensitivity in insulin‐resistant obese mice while stimulating GLP1 and insulin secretion. PAHSAs also augment insulin stimulated glucose uptake and Glut4 translocation to the plasma membrane in adipocytes. PAHSAs suppress inflammatory processes in immune cells. Biological effects of PAHSAs are mediated through lipid‐responsive GPCRs. In summary, we identified a novel lipid class that improves glucose‐insulin homeostasis. In conclusion, restoration of the low PAHSA levels in insulin‐resistant people may be effective to treat type‐2 diabetes.
Enhancer Transcription And Enhancer Function
Authors: Glass Christopher , Gosselin David , Link Verena , Romanoski Casey E. , Troutman Ty , Lam Michael , Eichenfeld Dawn
Published: November 2023

We utilize a combination of genome‐wide and genetic approaches to investigate molecular mechanisms that underlie the development and function of macrophages. Our studies suggest a relatively simple model of hierarchical interactions between lineage‐determining and signal‐dependent transcription factors that are required to select functional cis‐active regulatory elements from the genome. We find that signal‐dependent enhancer transcription is highly correlated with nearby gene expression and the writing of the enhancer histone signature at newly selected enhancers. In addition, we find that a subset of enhancer derived RNAs contribute directly to enhancer activity. We recently extended our approach from an in vitro analysis of macrophage activation to a systematic analysis of transcriptomes and enhancer landscapes of different tissue macrophage populations. We find that distinct tissue environments drive divergent programs of gene expression by acting on common enhancer elements as well as by directing the selection and activation of large sets of enhancers and super‐enhancers that are unique to microglia or resident peritoneal macrophages. Current efforts are exploring the impact of natural genetic variation on enhancer transcription and the relevance of enhancer transcription to cell specific gene expression in vivo.
How Different Are We? Performing Formalin Fixed Paraffin Embedded Tissue Immunohistochemistry Across Species.
Authors: Gibson‐Corley Katherine N. , Ofori‐Amanfo Georgina , Leidinger Mariah R. , Lambertz Allyn M.
Published: November 2023

The Comparative Pathology Laboratory (CPL) at the University of Iowa is affiliated with both a teaching hospital and a research university, which provides the opportunity to work with tissues from humans and a variety of research animals, including mice, rats, ferrets, rabbits, sheep and pigs. The CPL has developed procedures for immunohistochemical (IHC) antibodies on organisms for which the antibody has not previously been validated. In many instances, non‐standard research organisms are not referenced on antibody datasheets. Peer‐reviewed journal articles are referenced, but infrequently mention the organism and tissue we seek to study. The CPL utilizes Basic Local Alignment Search Tool (BLAST) software to compare antibodies against the organism of interest. BLAST comparisons between the two genetic sequences guide us to the antibody with the greatest probability of success. From years of experience, education, and experimentation, the CPL has developed guidelines for each animal of interest including the most successful blocking reagents and antigen retrieval methods. Since each antibody is unique, the CPL has also developed troubleshooting guidelines based on the IHC output which will be outlined. If a genetically similar antibody can be located, the CPL can create an IHC protocol using a limited number of variables to create a quality product: appropriate positive control tissue, antigen retrieval method, protein block, and primary dilution.

Support or Funding Information

Internal funding; Department of Pathology, University of Iowa
DNAJB1‐PKRACA ‐ A Novel PKa Fusion Protein That Drives Fibrolamellar Hepatocellular Carcinoma
Authors: Simon Sandy
Published: November 2023

Fibrolamellar hepatocellular carcinoma (FLHCC) is a rare liver tumor that usually occurs in adolescents and young adults. Characterized by hepatocytes with deeply eosinophilic, granular cytoplasm interspersed with fibrous bands, FLHCC is without signs of cirrhosis as in Hepatocellular Carcinoma. To characterize its molecular pathogenesis, we performed RNA‐seq and whole genome sequencing of FLHCC paired with normal liver from the same patient.

The majority of the DNA was unremarkable with few recurrent mutations or structural variants such as amplifications or inversions. The exception was a heterogeneous deletion of ~400 kB in one copy of chromosome 19. This produced a chimeric transcript and a fusion between the heat shock protein DNAJB1 and the catalytic subunit of protein kinase A (PRKACA) which retained full enzymatic activity. This chimera has been found in every patient (n>200). Differential expression analysis of RNA‐seq revealed many changes in the gene expression in FLHCC. These were generally consistent between patients with FLHCC and different from HCC samples from the TCGA or published reports of other cancers. Some of the observed changes can directly be linked to increased activity of PRKACA. Other changes involve pathways known to participate in the initiation and progression of other cancers, which, along with an increase of PRKACA activity, represent targets for therapeutic intervention. Several of these targets are now being explored in cellular models, genetic, and PDX mouse models, as well as clinical trials.

Support or Funding Information

The Rockefeller University Center for Clinical and Translational Science Grant 2UL1RR024143, The Sohn Foundation, The Rockefeller University, The Fibrolamellar Cancer Foundation
Transformation of the Introductory Biology Experience for Biology Majors: Progress of the Northwest Bioscience Consortium RCN‐UBE
Authors: Kruchten Anne , Baumgartner Erin , Beadles‐Bohling Amy , Whitman Jeffrey , Duncan Jason , Kayes Lori , Kiser Stacey , Shriner Walter , Stavrianeas Stasinos
Published: November 2023

The Northwest Biosciences Consortium is a group of two‐year and four‐year college faculty at both public and private institutions in the State of Oregon. Funded by an NSF RCN‐UBE grant, this group is working towards creating modern, student‐centered, integrated, and investigative introductory biology experiences for all students aligned with the Vision and Change: A Call to Action mandate. In our third year of the grant we now report on the activities of this group including two recent faculty workshops to encourage broad faculty involvement in our project goals. At the 2015 Winter Institute, faculty were introduced to active learning techniques that would help foster an inclusive environment for our students. At the 2016 Winter Institute, faculty participated in the development of instructional modules and assessment techniques in core areas identified by faculty as essential areas in the study of introductory biology. In addition to the results of the Winter Institutes, we will report on obstacles we have encountered especially in the area of smoothing the transition for transfer students taking an introductory sequence at a two year college followed by advanced coursework in biology at a four year college and plans for professional development activities for faculty across our institutions to encourage the adoption of student‐centered pedagogies aligned with Vision and Change and collaboration with other networks such as NW PULSE, PKAL, and NABT who are engaged in similar work. As a long‐term goal, we are particularly interested in helping faculty develop scaleable classroom modules and assessment practices that will encourage consistency in introductory biology outcomes across our diverse two‐year and four‐year institutions.

Support or Funding Information

Supported by the National Science Foundation RCN‐UBE grant (DBI‐1248121).
Original Course‐Based Undergraduate Research Experience to Detect Parasitic Worms in Local Parks in Two Undergraduate Courses
Authors: Schmidhauser Carol A , Blatch Sydella A
Published: November 2023

Traditional “cookbook” labs are a missed opportunity for students to participate in novel research and may not strongly reinforce skills needed for or interest in laboratory research. Studies have shown that course‐based undergraduate research experiences (CUREs), where students conduct original research as part of a traditional course, can increase student persistence in science and understanding of the scientific process. Many course‐based research projects either focus on experimental questions to which the answers are already known, or rely on methods that the instructor has already successfully applied to very similar experimental questions. This course‐based research experience differed in that experimental outcomes were not known in advance, and the instructors had little experience with significant portions of the methods used. Thus, students were almost entirely responsible for researching and developing the methods they needed to try. The students were given only their experimental aim and a brief overview of the methods that could be used. The overall aim of the experiment was to identify the presence of Toxocara spp. and Toxoplasma gondii worms in the Baltimore MD area. Parasitology students were tasked with collecting soil samples from a number of local parks, and isolating and identifying the parasite eggs from soil samples using floatation methods. Animal Physiology students were tasked with detecting parasite genes in DNA isolated from soil using PCR. It was the responsibility of the students to locate relevant primary literature, use the literature to propose specific methods, develop experimental controls, determine the materials and equipment needed, perform the methods, analyze the outcomes, and return to the literature to refine unsuccessful methods. Hypotheses developed from observations on this CURE will be tested in future experiments to examine effects of the experience on specific, measured outcomes of laboratory and quantitative skills, and attitudes towards science. Use of this kind of laboratory research question could easily be adapted to a CURE in biology courses of many types, as the presence of any kind of organism can be detected from a host of environmental samples.
The Design And Impact of a New Biomedical Seminar Deconstruction Course on Undergraduate Learning‐A Pilot Study
Authors: Osborne Sally , Schroeter Sara
Published: November 2023

We designed and piloted a research seminar deconstruction course to expose second year undergraduate students to high impact research, bridge the gap in their understanding of the scientific process and provide the opportunity to formulate their own research proposals. Deconstruction sessions included small and large discussion groups using flipped model of classroom teaching facilitated by problem sets with strong emphasis on web‐based resource library developed specifically for each seminar. Students were assessed based on written mini‐journals, seminar summaries, problem sets, and the oral presentation of their research proposal. Didactic sessions on guidelines for scientific presentation and writing research proposal were built‐in the course. Semi‐structured interviews and pre and post course surveys administered to ten students registered in the inaugural course show key learning gains in written and oral presentation of scientific material, interpretation of scientific results, understanding of the scientific process, confidence in developing realistic scientific research questions. The key challenges reported by students included coping with the high workload and planning their research proposal. The majority of the students reported learning skills that were transferable to other classes highlighting time management and critical thinking skills which they felt would enable them succeed in upper level research streams; some even described it as their favorite course.

Support or Funding Information

UBC Centre for Teaching, Learning & Technology
Navigating from Vision to Change: Tools to help biology departments align curriculum with the core concepts of biology
Authors: Brownell Sara , Wright Christian E , Couch Brian , Crowe Alison , Freeman Scott , Wenderoth Mary Pat
Published: November 2023

Vision and Change in Undergraduate Biology Education outlined five core concepts intended to guide undergraduate biology education: 1) evolution; 2) structure and function; 3) information flow, exchange, and storage; 4) pathways and transformations of energy and matter; and 5) systems.

We have taken these general recommendations and created the BioCore Guide—a set of general principles and specific statements that expand upon the core concepts, creating a framework that biology departments can use to align with the goals of Vision and Change. We used a grassroots approach to generate the BioCore Guide, beginning with faculty ideas as the basis for an iterative process that incorporated feedback from more than 240 biologists and biology educators at a diverse range of academic institutions throughout the United States. The final validation step in this process demonstrated strong national consensus, with more than 90% of respondents agreeing with the importance and scientific accuracy of the statements.

We are using the BioCore Guide to create a general biology programmatic assessment called Gen‐BioMaps that biology departments could use to monitor the progression of student understanding of the core concepts as they move through the major. Gen‐BioMaps uses a format of multiple T/F questions that span the sub‐disciplines of biology to test student understanding of the core concepts outlined in the BioCore Guide. We have implemented an iterative process in developing the stems and statements, including (1) revising based on student feedback from over 150 think aloud interviews, (2) piloting to over 1900 students nationally, and (3) incorporating expert feedback. We are currently revising the assessment and will be performing a final set of interviews, expert validation, and administration of the final version of GenBio‐MAPS in the spring of 2016.

Biology departments can use the BioCore Guide and Gen‐BioMaps as resources to guide curricular design, including helping faculty to articulate learning objectives aligned with Vision and Change, develop a more cohesive undergraduate biology curriculum, identify current gaps in the curriculum, and monitor student understanding of the core concepts of biology. It is our hope that both the BioCore Guide and the Gen‐BioMaps programmatic assessment will serve as agents of change for biology departments as we move toward transforming undergraduate biology education.

Support or Funding Information

NSF DUE 1323010
Modeling Aortic Insufficiency in the Scranton Cardiovascular Model
Authors: Graham Michelle Theresa
Published: November 2023

The effects of aortic insufficiency (AI) on cardiovascular hemodynamics are well known. Mean arterial pressure (MAP) decreases, driving compensatory mechanisms to reestablish MAP within a normal range. However, there are few pedagogical models designed to address instruction in the cardiovascular ramifications of valvular dysfunction. The Scranton Cardiovascular Model (SCM), a computer‐driven, fluid pumping, mechanical model of the cardiovascular system, was developed to improve cardiovascular physiology education1. We have extended the capabilities of the SCM to enable modeling of the hemodynamics of valvular dysfunction. A new component was fabricated to allow the normal SCM aortic valve to be quickly and reversibly replaced by a dysfunctional aortic valve. Insufficient valves of quantifiable regurgitant orifice area were designed and implemented to demonstrate systemic effects of progressive dysfunction, comparable to clinical data. The capability to model acute uncompensated AI and acute compensated AI was demonstrated through analysis of cardiac parameters such as mean arterial pressure, cardiac output, pulse pressure, and left ventricular end diastolic pressure. To simulate acute compensated AI, total peripheral resistance (TPR), contractility, and central venous tone were manipulated in the SCM until normal MAP was reestablished. Corresponding forward cardiac output (CO) reflected cardiovascular system efficacy. In uncompensated severe AI, MAP and CO, respectively, decreased to 66% and 73% of basal values. Multiple compensatory settings successfully reestablished normal MAP. The magnitude of the individual contributions of TPR and contractility to the restoration of MAP differentially impacted CO. Increased TPR restored MAP while decreasing CO. Increased contractility restored MAP while improving CO. We continue to fine‐tune SCM AI to more closely match clinical data. This “proof of concept” opens up a new pedagogical application for the SCM.

Support or Funding Information

Supported by the University of Scranton
Novel Graduated Engagement Strategy Tools for Enhancing Student Success in a Large Diverse Introductory Physiology Course
Authors: Malmquist Sarah J , Collins William F
Published: November 2023

In order to meet the need for both a STEM‐competent workforce, and a STEM‐literate electorate, efforts must be made to increase retention and performance of students from groups historically underrepresented in STEM, including women, low‐income students, and minority groups. Active learning strategies have been shown to narrow the achievement gap for many of these students, but the effects of specific strategies are relatively understudied. In addition, recent studies have shown improvements in student success in courses using alternative formats (e.g., flipped classes), although these gains may be due to the inherent inclusion of active learning, rather than a result of the flipped format itself. To address these questions, we are developing a novel active learning strategy, Graduated Engagement Tools (GET), that has multiple points for student engagement and utilizes (1) online and in‐class activities of increasing levels of Bloom's taxonomy and (2) directed metacognitive exercises to promote student self‐examination of strategies and motivations for learning. We are in the process of testing the general effectiveness of the GET strategy in an introductory physiology course with a very large (~1300), diverse student enrollment. Our initial focus is on improving content mastery and reducing failure rates. We will assess the effectiveness of these strategies across groups of students to measure differential effects on specific demographics of students. We also plan to investigate the effect of the GET strategy on self‐efficacy, or student's perceived ability to succeed; a metric shown to correlate with academic success and retention that is relatively low in students from groups historically underrepresented in STEM. These efforts are ongoing and we present examples of the GET activities, preliminary data showing high levels of student engagement and critical thinking, and a plan for creating and assessing a scalable and transferrable model for deploying these activities in both lecture and hybrid‐ / flipped‐classroom formats.
Fabric Muscles: A New Approach to Learning and Understanding Human Skeletal Muscles
Authors: Mathis Karen M.
Published: November 2023

Cat dissection is standard practice in Anatomy & Physiology classes. Use of this methodology to learn the skeletal muscles has several drawbacks: removal of the skin is time consuming and not useful; dissection of muscles is difficult for inexperienced students; muscle orientation is different from that of human's and muscles between species may or may not correlate. To help students better understand location, size and relative placement of human muscles, representations were created out of fabric and attached to human models. A survey of the students indicated that using fabric helped them to better understand the location and interaction between muscles in the arm (54%), leg (58%), face (59%), chest (66%), back (68%) and abdomen (73%), as well as the arrangement of muscles relative to each other (73%) compared to utilizing two dimensional drawings and three dimensional plastic models. In addition, 41/43 students believe this model should continue to be utilized in the future. Student enthusiasm and perceived learning were very high, so this model will continue to be implemented in forthcoming A&P classes.
Examining the immunomodulatory effects of IL‐12 in a mouse model of Alzheimer's Disease
Authors: Klaidman Lori , Ellefson Dolph D , Gallaher Tim , Stark Elena , Schettler Stephen P
Published: November 2023

Interleukin‐12 (IL‐12) is known to be a differentiation factor for naïve T‐cells into Th1 cells. We have previously used IL‐12 as a systemic therapeutic agent to clear Amyloid Beta (Aβ) peptides in a transgenic APPswe/PS1dE9 mouse model for Alzheimer's Disease. It was found that a modest decrease in Aβ plaque formation in this model using IL‐12 was associated with increased microglial activation using the marker, Ionized calcium‐binding adaptor molecule 1 (Iba1), also known as Allograft inflammatory factor 1 (AIF‐1). Iba1 is known to be expressed on activated microglial cells during an inflammatory response in the brain. In this study, we examined whether additional consecutive doses of IL‐12 would further magnify microglial activation in the brain. At 18 months old, wild type or AD mice were injected subcutaneously with IL‐12 once a week (20 ng) and sacrificed at day 21 or 42 (3 weeks vs 6 weeks). Mice were divided into the following groups: 1) WT mice, 2) WT mice with IL‐12 (3 weeks), 3) WT mice with IL‐12 (6 weeks) 4) AD mice, 5) AD mice with IL‐12 (3 weeks), and 6) AD mice with IL‐12 (6 weeks). Mouse brains were perfused and brain slices were quantitated by immunohistochemical (IHC) methods using imaging software for Aβ 1–40 or Aβ 1–42 peptides in AD brains, and for Iba1 in all groups. We further quantitated IL‐12rβ2 (IL‐12 receptor) using similar IHC techniques, in order to examine whether systemic IL‐12 administration also resulted in upregulated IL‐12 receptors in the brain. The results indicate a complex neuromodulatory effect.

Support or Funding Information

UCLA Department of Pathology and Laboratory Medicine
Neuritin, a New Potential Target for Optic Nerve Injury Therapy
Authors: Xu Jiajun , Cao Wenluo , Liu Weiye , Ma Haoqian , Qi Aiqing , Huang Tingting , Li Lin , Liu Fang
Published: November 2023

Optic nerve, which also belongs to the central nervous system, is difficult to repair once get injured. Neuritin, a novel member of the neurotrophic factor family, serves as a common downstream effector of neurotrophic factors and neural activity. Our experiment investigated the spatiotemporal expression of neuritin in adult rat retina and optic nerve in normal and after optic nerve crush. Following crush, neuritin protein and mRNA transiently increased in retina and optic nerve. In vivo, after neuritin lentivirus infection of rat retina, the number of retinal ganglion cells and optic nerve axons were increased significantly after injury, this profective effects was further confirmed by the functional tests like pupillary light reflex and visual evoked potential, suggesting that neuritin play a critical role in optic nerve regeneration. Further more, our results showed exogenous rhneuritin promotes RGC5 cell line survival and neuritin overexpression promotes neurite outgrowth which may through the upregulation of phosphorylation of Akt and Erk, and reduces cell apoptosis which may through the inhibition of caspase 3 activation after RGC5 damage in vitro. Our experiments suggested that neuritin is a potential new target for optic nerve injury therapy and provide alternative way for repairing injury of central nervous system.

Support or Funding Information

National Natural Science Foundation of China (Grant No. 31271282)
Accuracy of localisation and cause of preclinically suspected diagnoses. Explorative analysis of retrospective data of 700 patients
Authors: Hirtler Lena , Huber Florian
Published: November 2023

Introduction

The preclinical acute patient management by non‐physicians (i.e. paramedics, EMTs, etc.) is known to have large‐scale differences in accuracy and specificity, depending on numerous variables, such as rural vs. metropolitan regions or international differences in education of paramedics. The primary aim of our study was to find out differences in accuracy of anatomic localization and etiologic cause of preclinically suspected diagnoses between traumatic and atraumatic patients.

Patients & Methods

Seven‐hundred patients were randomly chosen as cases from the archived protocols of 2014 of a large city emergency medical service. The patients were assorted into two same‐sized groups, “atraumatic” and “traumatic”, which was performed depending on the designated clinical department. To find out suspected diagnosis, symptoms and respective anatomic region, and to match these with the eventual diagnosis, patient data was retrieved from EMS protocols and hospital reports. Statistical analysis of the two groups was performed and compared. We also evaluated side‐parameters such as presence or absence of intensified trauma training of transport leading paramedic and others.

Results

Preliminary results showed differences in the accuracy of anatomic localization and cause of preclinically suspected diagnoses in traumatic and atraumatic patients. Especially anatomic localization is more precise in traumatic patients than in the other collective. There was no significant increase in anatomic accuracy in traumatic patients who were treated by paramedics with certified special trauma course skills, compared to patients who were evaluated by staff without intensified EMS education.

Discussion

The effect of intensified trauma training is known to have had an impact on quality of practical skills in preclinical and hospital settings. It is discuss, whether and how training in anatomy and internal medicine could improve the accuracy and quality of patient evaluation by non‐physician EMS workers.

Support or Funding Information

The authors have nothing to declare.
How do Alternative and Traditional Dissemination Metrics Compare in Medical Education Scholarship?
Authors: Ramnanan Christopher J , Ambacher Kristin , Amath Aysah , O'Brien Cameron , Wood Timothy J , Leddy John J
Published: November 2023

Purpose

Medical educators need to demonstrate the dissemination of scholarship for academic promotion. Dissemination impact has traditionally been measured using citation‐based metrics. In recent years, other platforms (such as social media tools like Twitter and Facebook) have provided new avenues for authors to promote their work to both scholarly and non‐scholarly audiences. While alternative metrics (altmetrics) can capture non‐traditional dissemination data such as attention generated on social media, the academic value of altmetrics in medical education is unclear. The aim of this study was to determine which altmetrics are indices of access counts and citations in a highly cited, general interest medical education journal.

Methods

A database study was performed (August 2015) for all Medical Education papers in 2012 (n=236) and 2013 (n=246). Citation, altmetric and access (HTML views + PDF downloads) data were obtained from Scopus, the Altmetric Bookmarklet Tool, and the journal Medical Education, respectively. Correlation coefficients (r values) between variables were determined and statistical significance was calculated.

Results

Facebook, Google+, Reddit, and LinkedIn were each only used in the dissemination of a small fraction of papers and were not characterized further. Twitter and Mendeley were the only Altmetric‐tracked platforms utilized in the dissemination of greater than 50% of articles. For access counts versus Mendeley downloads, Twitter mentions, and Altmetric Scores, the correlation coefficients were r = 0.79, 0.56, and 0.52 for 2012 papers and 0.65, 0.32, and 0.35 for 2013 papers, respectively. For citation counts versus access counts, Mendeley downloads, Twitter mentions and Altmetric Scores, the correlation coefficients were r = 0.77, 0.81, 0.57, and 0.58 for 2012 papers and 0.62, 0.61, 0.18, and. 0.22 for 2013 papers, respectively. All correlations were statistically significant (P<0.01).

Conclusions

Mendeley downloads appear to be the strongest altmetric index of both readership and citations for articles in the high‐impact, general interest journal Medical Education. We conclude that access‐based indices (i.e. views and downloads) may be the best Altmetric tool to use in conjunction with citations for the purposes of measuring the impact of medical education scholarship.
X‐perience – an Interactive Viewing Platform Displaying Radiographic Profiles of Cadavers for Educational Purposes
Authors: Riogh Eithne Nic an , Perry Gary F , Jones James FX , Tattersall Adam , Heduan Martin , Stern Richard H , MacMahon Peter , Giannotti Nicola , Davis Michaela
Published: November 2023

Traditionally anatomy teaching has involved both learning from cadaveric dissection and formal lectures. The aim of our project was to introduce another dimension to students’ experiences of learning anatomy through the medium of radiology.

This project presents donor specific radiographic profiles of University College Dublin's cadavers for educational purposes. An interactive viewing platform Xperience was developed to vividly display these non‐generic images. Comprehensive donor profiles were created by adding medical histories and consultant radiologists’ reports. These self‐guided image‐based clinical cases enhance anatomical learning and offer early introduction of clinical radiography. X‐perience complements the dissection process by reinforcing the concept of the donor as the student's first patient.

Full skeletal radiographs were obtained from 13 donors (10 Female, 3 Male) aged 58–101 years. Radiographic images were produced digitally, labelled and stored using unique identifiers. These images were systemically labelled for normal and pathological findings and integrated with known medical history. Articulate Storyline 2 was used to build X‐perience, a HTML5 interactive interface. To assess the value of X‐perience Stage 3 Locomotor Biology students were surveyed. This cohort (n=50) had prior traditional anatomical teaching and could compare anatomical learning with and without X‐perience.

The workflow of profiling cadavers and constructing X‐perience proved successful. X‐perience is currently used by students during Dissecting Room practical classes and can be revisited via Blackboard, an online learning management system. Results of student acceptability survey, Kirkpatrick Level‐1, are extremely positive. 84% of students agree that X‐perience is relevant, easy to use and provides an integrated view of the human body. 88% appreciated the clinical relevance that X‐perience offers. 92% of students acknowledged that they now had a greater understanding of the importance of radiology.

The introduction of similar viewing platforms in other medical schools is recommended. Imaging modalities such as CT and/or histology could further refine X‐perience.

In addition to its educational value, X‐perience strengthens the body donation program, highlighting the individuality and dignity of each donor and generating enthusiasm amongst students.
FPR1 and Activation of Intestinal Fibroblasts
Authors: Reaves Titus A. , Anderson‐Thomas Meghan , Nillas Andrea , Lecher Mackenzie
Published: November 2023

Fibroblasts are mesenchymal cells that produced the connective tissue consisting of collagens, laminins, and proteoglycans. Myofibroblasts are activated fibroblasts that express α‐smooth muscle actin and contract during wound healing. If activation continues, myofibroblasts become over‐activated and intestinal fibrosis (excessive and dysregulated collagen production that affects the function of the intestine) may result. Despite this information, the molecular events of fibrosis are not completely understood. We have shown that fibroblasts express the Nα‐formyl‐L‐methionyl‐L‐phenyalanine receptor (FPR1), which is a G‐protein coupled receptor that regulates the leukocyte response to bacterial formylated peptides. We initially exposed intestinal fibroblasts to a variety of inflammatory mediators that include lipopolysaccharide (LPS), IL‐6, and IL‐8. Interestingly, LPS treatment resulted substantial upregulation of FPR1, while IL‐8 treatment resulted in a substantial down‐regulation of FPR1. Using Flow Cytometry, similar results were obtained. We examined whether constitutive activation of the FPR1 (and inhibitors) resulted in the over‐activation of myofibroblasts and ultimate release of excessive and dysregulated collagen. Studies revealed that reducing FPR1 exacerbates the inflammatory response of fibroblasts. Taken together, the FPR1 appears to modulate the activation of fibroblasts. Down‐regulation of the FPR1 may lead to a better understanding the fibrotic response and better treatments of dysregulated inflammatory conditions involving fibroblasts.

Support or Funding Information

National Science Foundation RII
CD43 contributes to mouse Th17 cell adhesion to intercellular adhesion molecule‐1
Authors: Velazquez Francisco Esteban , Salvador Ane , Nevers Tania , Alcaide Pilar
Published: November 2023

CD43 is a sialoglycoprotein with many functions and roles that can serve as a pro‐adhesive and anti‐adhesive molecule. In T cells, CD43 has been shown to function as an E‐selectin (E‐sel) ligand in cooperation with PSGL‐1. We recently demonstrated that CD43 functions as an E‐sel ligand for T helper 17 (Th17) cells, which participate in autoimmunity and in chronic inflammatory reactions when recruited at sites of inflammation. Moreover, CD43−/− mice are protected from Experimental Autoimmune Encephalomyelitis (EAE) and have decreased Th17 cell recruitment into the spinal cord. Interestingly, E‐sel is not required for the development of EAE, and other adhesion molecules such as intracellular adhesion molecule 1 (ICAM‐1) and vascular cellular adhesion molecule 1 (VCAM‐1) mediate T cell recruitment in EAE instead. Despite the subset specific use of CD43 as an E‐sel ligand by Th17 cells, other potential pro‐adhesive and anti‐adhesive properties of CD43 on Th17 cells to adhesion molecules such as ICAM‐1 and VCAM‐1 have yet to be properly characterized. We hypothesize that Th17 cells use CD43 as a pro‐adhesive molecule to adhere to other adhesion molecules and infiltrate the spinal cord in EAE in an E‐selectin ligand independent manner. We use real‐time live video microscopy under flow conditions, Th17 cells generated in vitro from WT and CD43−/− mice, and the experimental autoimmune encephalomyelitis (EAE) model of autoimmunity and inflammation. We report that upon immunization with myelin oligodendrocyte glycoprotein (MOG), ICAM‐1 was upregulated in the spinal cord of WT and CD43−/− mice, in contrast with E‐sel, which had undetectable levels as determined by qPCR. Additionally, we observed that CD43−/− Th17 cells have impaired adhesion to ICAM‐1 in vitro, but express similar levels of LFA‐1, the main ICAM‐1 ligand in T cells, as WT mice. Moreover, WT and CD43−/− Th17 cells adhered to ICAM‐1 in detachment assays with similar strength. Our results demonstrate the possibility of CD43 interacting with ICAM‐1 to mediate Th17 cell recruitment to sites of inflammation. Future studies will unveil whether direct interaction between CD43 and ICAM‐1 occurs specifically in Th17 cells as a mechanism of Th17 cell recruitment to sites of inflammation independent of E‐sel, such as EAE.
Optimization of DNA, RNA and RNP Delivery Methods for Efficient CRISPR/Cas9 Mediated Mammalian Cell Engineering
Authors: Khodthong Chuenchanok , Snow Josh , Juckem Laura
Published: November 2023

The CRISPR/Cas9 genome‐editing platform is a versatile and powerful technology to efficiently create genetically engineered living cells and organisms. This system requires a complex of Cas9 endonuclease protein with a gene‐targeting guide RNA (gRNA) to introduce double‐strand DNA breaks (DSBs) at specific locations in the genome. DSBs are then repaired by the error‐prone Non‐Homologous End Joining (NHEJ) pathway, resulting in insertions and/or deletions (indels) which disrupt the targeted locus.

The success of CRISPR genome editing experiments is limited by the intracellular delivery and expression of Cas9 protein and gRNA. Many methods for achieving CRISPR mediated cleavage have been identified and the choice of DNA, RNA or ribonucleoprotein (RNP) format is dictated by experimental goal and cell type. Transfection of each type of molecule requires specific considerations for efficient functional delivery. We performed transfections using different combinations of molecules including: plasmid DNA, messenger RNA, Cas9 protein and gRNA to maximize targeting of the Cyclophilin B (PPIB) gene in HEK 293T/17, U2OS, and other mammalian cell types. Our results extend the utility of the CRISPR/Cas9 system by identifying optimal transfection conditions for intracellular delivery of Cas9 and gRNA in different formats.
Design and construct a type of fluorescently labeled circular DNA molecules to study DNA topology and topoisomerases by fluorescence resonance energy transfer
Authors: Leng Fenfei , Gu Maxwell
Published: November 2023

DNA topology, an essential property of the DNA double helix, is under tight homeostatic control in vivo. Studies of DNA topology and topoisomerases are critical for understanding the mechanism underlying several infectious and genetic diseases and also for designing new anti‐cancer and anti‐microbial drugs targeting DNA topoisomerases. Typically agarose gel electrophoresis is used to study DNA supercoiling and topoisomerases. However, gel electrophoresis is time‐consuming and labor‐intensive. It is desirable to develop other assays, such as fluorescence‐based assays, for such studies. In this article, we developed a new method to construct a type of fluorescence‐labeled circular DNA molecules with high yields to study DNA topology and topoisomerases by using fluorescence resonance energy transfer (FRET). Specifically, we successfully produced relaxed and supercoiled pAB1_FL905 that carries fluorescein and dabcyl labels in the 42 bp AT sequence. Our results showed that pAB1_FL905 is a powerful tool to study DNA topology and topoisomerases by FRET. pAB1_FL905 can also be developed into rapid and efficient high‐throughput screening assays to identify inhibitors from the millions of compounds found in small molecule libraries that may target DNA topoisomerases, such as bacterial DNA gyrase and human DNA topoisomerase I and II.

Support or Funding Information

1R15GM109254‐01A1 from the National Institutes of Health
Genomic Analysis of the Resistance of E. coli to Benzyl Bromide
Authors: Qi Macee , Muenter Mark , Baig Samir , Bellou Sirine , Carlson Alyssa , Conway Charles , Joshi Christopher , Kramer Caitlin , Leifer Becky , Travers Meghan , Wang Poguang , Giese Roger , Beuning Penny
Published: November 2023

Exposure to an alkylating agent can occur via industrial exposure or treatment in cancer chemotherapy as cytotoxic agents, causing DNA alkylation. DNA alkylation is a common occurrence in which alkyl groups are covalently attached to DNA bases resulting in DNA lesions. DNA lesions can arise either from endogenous sources through metabolism or exogenously from environmental exposure. The resulting lesions can cause mutations that can lead to cancer. Therefore understanding the DNA alkylation process and identifying the major DNA adducts that form from specific alkylating agents is important. The overall aim of our research is to determine the survival of E. coli AB1157 cells when treated with benzyl bromide (BnBr), an alkylating agent used in the chemical and pharmaceutical industries. The survival of wild‐type E. coli AB1157 and twenty‐seven derivatives, each harboring a knockout of a specific gene associated with various DNA repair processes has been determined upon exposure to the alkylating agent BnBr. In order to confirm whether the deletion of specific genes in E. coli that showed resistance when exposed to the alkylating agent is indeed the cause of decreased survival, complementation studies are being carried out. Benzylation profiles for free nucleosides (dA, dT, dC, dG) reacted with BnBr have been determined using HPLC‐MS. Analyses of various DNA samples exposed to BnBr are being carried out to characterize the major adducts formed, allowing us to determine how DNA repair pathways contribute to repairing the specific adducts.