bluuebunny/crossref_metadata_2025 huggingface dataset:
A Metric of RNA Quality for RNA Isolated from Formalin Fixed, Paraffin Embedded Tissue
Authors: Hewitt Stephen , Chung Joon‐Yong , Perry Candice , Ylaya Kris
Published: June 2021

RNA isolated from formalin fixed, paraffin embedded tissue is now commonly used in both investigative and diagnostic pathology. Extensive research has demonstrated that although RNA is present in paraffin embedded tissue, it is subject to degradation from both fixation and impregnation protocols as well as storage. The extent of degradation is highly variable, and has direct impact on what assays can be performed, as well as the quality of results. Although the RIN (RNA Integrity Number) is widely applied to RNA isolated from paraffin embedded tissue, this ratio of rRNA peaks obtained from the electrophoretogram, is a poor indicator of RNA quality from paraffin embedded tissue, especially considering that these peaks are frequently absent in these samples. Using the same platform to evaluate RNA integrity, we have developed a novel metric, called the QM (Quality Metric) to quantify RNA quality from formalin fixed, paraffin embedded tissue. The metric is based on a weighed area‐under‐the‐curve approach. This metric can be applied to RNA obtained from any paraffin embedded tissue, and used as a quality metric for evaluation of fixation studies, impregnation protocols, studies of block storage as well as RNA isolation protocols. Additionally the QM can be used to qualify RNA for different end‐point studies, including RT‐PCR and branched‐DNA based assays.
Destabilization of endothelial cell‐cell contacts modifies inflammatory responses
Authors: Glading Angela , Corr Maripat , Lerman Irina , Sarelius Ingrid , Kuebel Julia
Published: June 2021

Changes in the stability of endothelial cell‐cell contacts are commonly associated with chronic and acute vascular inflammation. However, it remains unclear how changes in endothelial cell‐cell contact impact the progress of vascular inflammation. KRIT1‐deficient animals lack the adherens junction accessory protein KRIT1, and exhibit a loss of cell‐cell contact stability. Krit1+/‐ mice exhibit increased baseline microvessel permeability, but maintain normal leukocyte responses. Stimulation of Krit1+/‐ mice with various inflammatory mediators enhanced edema formation but not leukocyte activation. Krit1+/‐ mice exhibited relatively normal leukocyte responses to TNF‐α, IL‐1β, and histamine, but, notably, Krit1+/‐ vessels did not become more permeable following TNF‐a treatment, contrary to what was seen in littermate controls. Though TNF‐α signaling in KRIT1 deficient endothelial cells appeared normal, these cells displayed increased oxidative stress, and treatment with TNF‐α failed to enhance reactive oxygen species (ROS) levels. In vivo, targeted inhibition of endothelial ROS was sufficient to restore microvascular permeability in Krit1+/‐ mice to WT levels. However, antioxidant treatment of TNF‐α‐treated WT and Krit1+/‐ animals reversed only venular permeability. KRIT1 depletion‐induced oxidative stress could be reversed by treatment with an NADPH‐oxidase inhibitor, suggesting that TNF‐α driven induction of NADPH oxidase may be inoperative in the absence of KRIT1. Thus, destabilization of endothelial adherens junctions following depletion of KRIT1 plays a vascular bed‐dependent role in modifying the vascular response to inflammation.
Modulating Neutrophil‐Derived MPO‐Endothelial Surface Binding with CORMs
Authors: Patterson Eric , Fraser Douglas , Inoue Ken , Cepinskas Gediminas
Published: June 2021

Myeloperoxidase (MPO) binds to vascular endothelial cells (EC) through a mechanism that, at least in part, involves interactions with glycosaminoglycans expressed on the endothelial surface. MPO that is closely associated with endothelial cells can directly damage them through its catalytic activity. Our previous work demonstrated that carbon monoxide‐releasing molecule 3 (CORM‐3) inhibits MPO catalytic activity and can thereby mitigate its detrimental effects during severe inflammation. The effects and mechanisms of CORM‐dependent modulation of MPO‐vascular EC binding are not yet investigated.

We have employed in vitro assays to study neutrophil‐derived MPO binding to human umbilical vein EC (HUVEC) following treatment with two structurally different CORMs; CORM‐3 and CORM‐401, synthesized based on Ru or Mn transition metals, respectively.

Our results indicate that at equimolar concentrations CORM‐3, but not CORM‐401 inhibits MPO‐endothelial surface binding. Inactivated CORM‐3 (iCORM‐3) did not inhibit MPO binding to EC

We conclude that in addition to CORM‐3‐dependent suppression of MPO catalytic activity, reduced MPO binding to EC in the presence of CORM‐3 may provide an additional mechanism that contributes to CORM‐dependent suppression of inflammation and endothelial damage.
Update on the Development of Curative Cancer Immunotherapies
Authors: Restifo Nicholas
Published: June 2021

Invasive cancer results in the death of more than 8 million people worldwide and over have of these deaths occur in the developing world. With few exceptions, the most commonly used therapies – surgery, chemotherapy and radiation therapy – do not result in long term durable complete responses in patients once cancer has spread beyond its primary site. Over the past several years, cancer immunotherapy based on the adoptive transfer of a patients own (autologous) cells – be they naturally‐occurring or genetically‐engineered – is rapidly establishing itself as a viable approach to the treatment of metastatic cancer. Some patients have experienced complete responses ongoing beyond five years. Current efforts are aimed at understanding the T cell qualities that enable the induction of long‐term durable complete responses in patients. While drug‐based therapies last for hours or days, cell‐based therapies can last for weeks or months, even years. We have now identified the properties of human stem cell‐like anti‐tumor T cells that can effectively combat the stem cell‐like process of cancer. These efforts are akin to 'fighting fire with fire.' Adoptive cell transfer immunotherapy is a rapidly developing new approach to the therapy of metastatic cancer in humans and has mediated complete regression of cancers in patients with metastatic melanoma, refractory leukemias and lymphomas and in patients with advanced cervical cancer.
Immunomodulation Therapy goes to Tumor Site
Authors: Chen Lieping
Published: June 2021

The PD‐1/PD‐L1 (B7‐H1) immune modulatory pathway plays important roles in suppressing antigen‐specific immune responses and inflammation. Selective expression of B7‐H1 in tumor microenvironment and subsequent interaction with PD‐1 on T cells is demonstrated to be a major mechanism of losing T cell immunity in tumor sites in a significant fraction of cancer patients including lung cancer. Monoclonal antibodies blocking this pathway have been tested in >2,000 patients and induced significant regression in approximately 1/3 of patients with late‐stage cancers of lung, kidney, skin, bladder, head and neck and some lymphomas in multiple institutions in US. The treatment is well‐tolerated and the clinical responses are long‐lasting. In the context of ongoing large‐scale clinical trials in thousands of cancer patients, understanding of immunological mechanisms underlying response and resistance to anti‐PD‐1/PD‐L1 therapy is critical for further improvement of cancer therapy in the future.
Conformational Changes of a Type II ABC Importer for Small Substrate Transport
Authors: Pinkett Heather
Published: June 2021

ATP‐Binding Cassette (ABC) importers work as molecular pumps to transport substrates across the cellular membrane into the cytoplasm. To understand how the helices in the transmembrane domains (TMDs) of a transporter reposition themselves to allow a small compound to pass into the cell, our research has focused on ABC importers from Haemophilius influenzae as a model. Our studies of ABC importers revealed two transport systems within the same organism that transport the same substrate, molybdate. Using electron paramagnetic resonance spectroscopy (EPR) to capture a snapshot of the MolB2C2‐A complex at each point in the transport mechanism, we identified four states, from apo to post‐ATP hydrolysis. The translocation pathway of MolBC opens just enough to allow through a small substrate, suggesting that in addition to the SBP, the size of the translocation pathway size provides an additional selectivity filter; thus, the transport mechanisms of ABC importers are more mechanistically diverse than previously thought. These results reveal that the mechanism of a Type II transporter for a small substrate may differ when compared to the mechanism transporters of large substrates.
Entamoeba histolytica Induces Pro‐Inflammatory Mediator HMGB1 Release in Early Contact with Macrophage
Authors: Begum Sharmin , St‐Pierre Joëlle , Moreau France , Chadee Kris
Published: June 2021

Entamoeba histolytica (Eh) is an invasive extracellular parasite that causes amebiasis in ~10% of infected individuals resulting in 105 deaths/year. The host and/or parasite factors that influence disease susceptibility remain unknown. Interaction between Eh and host macrophages has been shown to trigger a strong pro‐inflammatory response characterized by IL‐1β and TNF‐α release. High Mobility Group Box1 (HMGB1) is a non‐histone nuclear protein secreted by macrophages upon stimulation with various bacterial, viral and cellular components. HMGB1 binds TLRs (TLR‐2 and 4) and induce pro‐inflammatory responses in neighboring cells. In this study, we investigated whether Eh could trigger the release of the pro‐inflammatory molecule HMGB1 as an early “alarmin” signal and the putative parasitic virulent factors involved. Only stimulation with live Eh but not Eh components triggered prompt (1‐5min) and robust secretion of HMGB1 in human macrophages. This event was contact‐dependent as inhibiting Eh adhesin, Gal‐lectin with exogenous galactose abrogated HMGB1 release. HMGB1 secretion was independent of the major surface virulent factor cysteine proteinase 5 (EhCP5) as CP5‐deficient Eh, or cysteine protease inhibitor pretreatment of Wt Eh had no effect. HMGB1 secretion was independent of caspase‐1 activation, as revealed by using pan caspase inhibitor, Z‐VAD‐fmk. Eh‐induced HMGB1 release upon contact with macrophage acts as a sensor for the host to detect invasive Eh that can attract bystander cells to amplify innate host defenses and/or promote an acute pro‐inflammatory response critical in disease pathogenesis.

Grant Support: NSERC
Suppression of Plk3 expression by Ni(II) may contribute to nickel carcinogenesis in the lung
Authors: Li Cen , Park Soyoung , Dai Wei , Xu Dazhong
Published: June 2021

Exposure to nickel (Ni) compounds is known to induce human lung malignancies. Research shows that Ni induces lung tumorigenesis mainly through epigenetic and metabolic pathways. Polo‐like kinase 3 (Plk3) is a protein kinase that has been implicated in lung tumorigenesis. In this study, we examined the effect of Ni on Plk3 in A549 lung carcinoma cells. We find that NiCl2 lowers the Plk3 protein level in time‐ and dose‐dependent manners. A549 cells treated with cycloheximide in the presence or absence of NiCl2 exhibited significantly reduced levels and the half‐life of the Plk3 protein. In addition, A549 cells treated with MG132 partially reversed the effect of NiCl2 on the Plk3 protein level. Our experiments also revealed that majority of the endogenous Plk3 protein is located in the cytoplasm where degradation of Plk3 appears to occur. Transfection of A549 cells with FLAG‐tagged ubiquitin (Ub) followed by treatment with NiCl2 and/or MG132, immunoprecipitation with an anti‐FLAG antibody, and Western blot with an anti‐Plk3 antibody showed that Plk3 was poly‐ubiquitinated and that the ubiquitination was significantly enhanced by NiCl2 and/or MG132 treatments. Furthermore, real‐time PCR analysis showed that the Plk3 mRNA level was also reduced by NiCl2 treatments. Together, our data show that NiCl2 suppresses Plk3 expression by lowering the PLK3 mRNA level and promoting proteasome‐mediated degradation of the Plk3 protein. Given the potential role of Plk3 as a tumor suppressor in the lung, suppression of Plk3 expression could be one of the mechanisms that contribute to Ni‐induced lung carcinogenesis and tumor progression.
Integrating Mathematics Principles in Cell Biology
Authors: Lin Danielle , Mimbs Debra , West Lori
Published: June 2021

Increasing integration of mathematics and biological concepts is an area of intense focus in education. Our goal is to develop a platform through which elementary mathematics and science instructors can collaborate with one another in order to demonstrate to their students the connectedness of the two disciplines. Our focus is to apply mathematics common core standards to biological phenomena. One strategy is to show how standards in geometry can be applied to cells. In order to do so, cells of various shapes were viewed under the microscope and then photographed. The images were incorporated into Geometer's sketchpad where perimeter, circumference, and/or area were determined based on the relative shape of the cell. Although this experiment was developed with elementary teachers in mind, we feel that extensions of this exercise would be well suited in secondary and higher education.
Having PhUn in Australia While on Summer Vacation
Authors: Halpin Patricia
Published: June 2021

When planning our family vacation to visit relatives in Australia, I looked into the opportunity to perform a PhUn Week activity there. PhUn Week is an annual outreach program in which K‐12 students get to learn about physiology through meeting a physiologist and performing an experiment. My cousin put me in touch with a teacher in a private parochial school that combines 3rd and 4th grade classes, and I taught two back‐to‐back sessions. I started by asking: “Does anyone know what a physiologist is?” Finding from one eager student that “physiologist” is confused with “physicist,” I went on to describe what a physiologist is, and then how the heart works, and then the lungs, and finally how the two organ systems work together. The students had recently done a unit on Scientific Method, and together we designed a hypothesis – “Exercise increases heart rate” – and a testing method in which students would measure their heart rates, run for 3 minutes, and measure their heart rates again. To ensure a steady exercise pace when we performed our experiment, I had my sons there to help – one advantage of being on a family vacation. Instructions to the class were, “Do not pass older son in front or fall behind younger son.” After data collection, students readily shared their results and concluded that they had proved their hypothesis. I then explained why the heart rate should increase during and after exercise. We also discussed healthy eating and why being active is beneficial. The experience was clearly effective and well worth the time away from the usual vacation activities. PhUn Week forms partnerships among educators and inspires K‐12 students to think about becoming scientists. I urge other physiologists to incorporate a little PhUn in their next vacation.
Leg Bone Geometry in Human Diabetic Neuropathy
Authors: Honig Helen , Allen Matti , Allman Brian , Rice Charles
Published: June 2021

Bone geometry is an important indicator of bone health and fracture risk, but has not been studied in individuals with diabetic neuropathy (DN). The objective was to investigate the effects of DN on tibial cortical and medullary cross‐sectional areas (CSA) using magnetic resonance imaging. Sequential images of 1mm thick slices were acquired of the right leg from the tibial plateau to the talus in 6 individuals diagnosed with DN and 8 age‐matched (32 to 79 y) controls. The CSA (cm2) was measured (average of 2 adjacent slices) at 3 sites, 20% (proximal), 50% (middle) and 80% (distal) of tibial length, by a blinded analyzer. At the proximal site only, medullary CSA in DN was significantly greater than controls (means + SD: 6.1±1.4 vs. 4.6±0.4). Also, as a percent of total CSA, DN compared with controls had significantly less cortical (~30% vs. ~38%) and greater medullary (~69% vs. ~62%) areas. At middle and distal sites there were no differences in any measures. These preliminary results indicate bone geometry is negatively affected by DN at the proximal tibia. This may be due to lower weight bearing or mechanical loading than at the middle or distal aspects. Presumed lower levels of physical activity in DN coupled with less muscle mass and strength, but heavier body weights, may be important factors influencing bone geometry to consider in future studies, including assessment of the lesser weight‐bearing fibula.

Supported by NSERC.
Distribution of Neural Factors and Angiogenesis Markers between Human Tensor Veli Palatini Muscle and Tensor Tympani Muscle
Authors: Sato Iwao , Miwa Yoko , Yatsu Toshio , Sunohara Masataka
Published: June 2021

Substance P‐immunoreactive nerve fiber (SP‐IR NF) and calcitonin‐gene‐related peptide‐immunoreactive nerve fiber (CGRP‐IR NF) are important mediators of neurogenic inflammation and blood supply. The presence of SP‐IR and CGRP‐IR NFs indicate vascular regulation, such as vasodilation or vasoconstriction (de Hozet al., 2008). SP‐IR and CGRP‐IR NFs have been found in mammalian middle ear mucosa (Ylikoski, 1989). However, the distribution of SP‐IR and CGRP‐IR NFs in the blood vessels of the human tensor tympani muscle (TTM) have not previously been reported. In this study, the TTM, tympanic membrane, malleus in the middle ear and tensor veli palatini muscle (TVPM) were examined by whole‐mount immunohistochemistry or cross‐section for distribution of cluster of differentiation 31 (CD31) and lymphatic endothelial marker (D2‐40) in tissue from Japanese subjects. Thirteen human cadavers (ranging in age from 46‐90 years) were used in this study. SP‐IR and CGRP‐IR NFs were primarily found on vessels at the origin, insertion and belly of the surface of two muscles. Moreover, different distribution of CD31 and D2‐40 were also detected using immunohistochemical methods. The SP‐IR and CGRP‐IR NFs neural factors were detected on the surface of the malleus and the insertion of the TVPM in our results. Different distribution of neural factors (SP‐IR and CGRP‐IR NFs) and angiogenesis markers (CD31 and D2‐40) were found in the cross‐section of two muscles. Therefore, our results indicate that existence of the SP‐IR and CGRP‐IR NFs of the TTM and the TVPM may reflect to characterize property of stimulation for pain or inflammation and also play different role of auditory muscular system in human.
Fungal Contamination of Plastined Specimen
Authors: Barros Mirna , Navarini Alessandra , Masuko Telma Sumie
Published: June 2021

Introduction

Plastination is a unique technique of tissue preservation developed by Gunther von Hagens in 1978. In this process, water and lipids in biological tissues are replaced by curable polymers (silicone, epoxy) which are subsequently hardened, resulting in dry, odorless and durable specimens. The advantages of use plastinated specimens are they can be conveniently stored, the tissues are safe to handle, toxic fixatives are eliminated, they are durable, having a much longer shelf life. This study aimed to demonstrate fungal contamination of a plastined kidney.

Material and method

On the surface of a plastined kidney divided in two parts, we observe a thin layer of a white substance, which is presented into more or less dense portions. Six samples of that substance from different places were collected, inoculated in Sabouraud‐agar and incubated at room temperature. The samples were checked dailly for the presence of fungi.

Results

We observed the growth of fungi in one sample on the eighth day after inoculation and the fungus was identified as Penicillium sp.

Conclusion

The foundation of all good tissue specimen preparations is complete fixation. The primary function of tissue fixation is to prevent putrefaction and autolysis. Probably, the fixation of the specimen was not completed and during storage, the fungal contamination occurred. We can not rule out the possibility of contamination of solutions used in the process of fixing and plastination specimen.
Effect of Cadaver Dissection on Written and Practical Gross Anatomy Test Scores
Authors: Olivieri Nina , Hurley Mary , Olivieri Marion
Published: June 2021

Over the past five years, 325+ Italian medical students participated in D'Youville College's (DYC's) Giovanni Mazzotti Anatomy Conference (GMAC), named in honor of the late president of the Medical School of Bologna and the Italian Anatomy Society. Cadaver dissection is not common in Italy where models, drawings and computer‐generated images are utilized. Some universities select participants for anatomy competency.

Before they dissect, attendees complete 50 question practical and 50 question written examinations (choice of English or Italian). After ~100 hours of available laboratory time, equivalent examinations are completed and compared. To quantify anecdotal proclamations of greater knowledge of anatomy, in this preliminary study, we examined 107 students from different populations (Group A:47, Group B:60). On average, a significant improvement was observed in test scores increasing by 5% in group A and 10% in Group B with p 蠄 0.001) in basic written and practical knowledge of anatomy. Students with the weakest practical pre‐test scores made the largest advances by conference end with as high as a 42% increase. Cadaver dissection and experience was found to be beneficial and worthwhile in increasing the practical as well as theoretical knowledge of these medical students.

We expect this collaboration to continue and will include exam modifications and additional participant information to provide a more in depth analysis. We thank our friends and collaborators, Professors Zecchi, Manzoli, Nori, Papa and Sette of the Medical Universities of Florence, Bologna, Salerno, Naples, and of Rome Tor Vergata. Supported by the GMAC and approved through the IRB of DYC.
Dissection in Gross Anatomy Course; The Need for a Learner‐Centered Pedagogy
Authors: Musa Muhammad , Bello Abubakar , Abdulsalam Ali , Bala Garzali , Onimisi Bethel , Ammani Tijjani
Published: June 2021

In Nigeria and as far as the medical curriculum is concerned, dissection of cadaveric bodies in gross anatomy course is one of the most important activity in the teaching and learning of anatomical sciences. Dissection, if properly conducted is believed to impact meaningfully in the future generation of medical doctors and professional anatomists especially as it widens their knowledge base and further enhances their basic understanding of clinical manifestations during the course of their training. However, evidences show that the most common methods of teaching used in gross anatomy dissections are teacher‐centered. This article reviewed some teaching methods employed by anatomy educators/demonstrators during dissection sessions in gross anatomy taking a close look at Nigerian Medical Schools. Alternative ways and strategies are discussed to further incorporate learner‐centered pedagogies for the purpose of achieving the goal of dissection in the gross anatomy course. Conclusively, the need has been identified for anatomy educators/demonstrators of various anatomy departments to employ learner‐centered approaches that will enhance and promote effective teaching / learning process during gross anatomy dissections.
The Constructive Approach to Teaching Anatomy through Clay Modeling: A Multi‐Tier Educational Value
Authors: Márquez Samuel , Patel Jasmine , Rosentsveyg Juliana , Márquez Anjelica , Pagano Anthony
Published: June 2021

The art of teaching anatomy has been traditionally performed via dissection. Most anatomy courses, ranging from fetal pig dissection in high school (HS) to human donor dissection in medical school, require a destructive process for discovering anatomical structures in 3D space. In contrast, clay modeling (CM) is a constructive approach to understanding spatial anatomical relationships. This qualitative study reports the experiences of two former medical students, a Syracuse HS teacher, anatomy faculty from two medical schools, and a third grader who participated in daylong workshops using the Anatomy in Clay Learning System. This haptic approach, used to model female perineal muscles from clay, emphasized touch and proprioception with continuous iterative feedback to ensure learning throughout the activity. While the instructors and students (now resident physicians) gained a greater appreciation of the region, a new world of anatomy was exposed to an unsuspecting, but curious, elementary school student. Participants with anatomy experience reported much improved familiarity with the pelvis, and in particular the attachments of the obturator internus muscle, even after having dissected the region previously. While the pedagogical question whether to implement CM before or after lab dissections arose, the experience revealed CM to be a highly valuable tool across multiple educational tiers.
Browsing Software of the Visible Korean Data Used for Teaching Sectional Anatomy
Authors: Chung Beom Sun , Chung Min Suk
Published: June 2021

The interpretation of computed tomographs (CTs) and magnetic resonance images (MRIs) to diagnose clinical conditions requires basic knowledge of sectional anatomy. Sectional anatomy has traditionally been taught using sectioned cadavers, atlases, and/or computer software. The computer software commonly used for this subject is practical and efficient for students but could be more advanced. The objective of this research was to present browsing software developed from the Visible Korean images that can be used for teaching sectional anatomy. One thousand seven hundred and two sets of MRIs, CTs, and sectioned images (intervals, one millimeter) of a whole male cadaver were prepared. Over 900 structures in the sectioned images were outlined and then filled with different colors to elaborate each structure. Software was developed where four corresponding images could be displayed simultaneously; in addition, the structures in the image data could be readily recognized with the aid of the color‐filled outlines. The software, distributed free of charge, could be a valuable tool to teach medical students. For example, sectional anatomy could be taught by showing the sectioned images with real color and high resolution. Students could then review the lecture by using the sectioned and color‐filled images on their own computers. Students could also be evaluated using the same software. Furthermore, other investigators would be able to replace the images for more comprehensive sectional anatomy.
Production of an Anatomical Video of the Middle Cranial Fossa and Its Conversion into an Interactive Teaching Module (550.12)
Authors: Oliver Peter
Published: June 2021

Videos are an excellent tool to present and introduce anatomy to students. They especially provide an overview that includes a three dimensional appreciation of the subject matter. A high resolution video of the middle cranial fossa was produced using Adobe software. The significant anatomical points of interest were described by narration. The video presentation included the boundaries and bony landmarks of the middle cranial fossa. Numerous blood vessels and nerves associated with the region are well illustrated. Camera angles, zooming and motion were implemented during the capture to enhance three dimensional appreciation of the viewer. In the present study, I have sought to extend the students learning experience to include an interactive self‐paced learning module of the subject matter. Images from the video were captured and placed in a sequence to so that the viewer can operate a timeline scrubber to advance the images at their own pace. They can do a quick overview of the images and then come back and focus on more detail. The images in the timeline are labeled with links that enact popups for access to more detailed descriptions and explanations of the of the item indicated in the image, e.g. information regarding the distribution of the ophthalmic division of the trigeminal nerve. The use of an interactive module should enhance the students attention and cater to the pace of the individual student. It is planned that such modules will be introduced within the context of their initial anatomy studies to augment student learning. The study modules are projected to be used as a review and refresher of previously learned material. This approach will use the same material for both initial learning and subsequent review.
Development of an Online Tool for Periodontal Disease Education
Authors: Johnston Sari , Al‐Osman Abdul , Kalman Les , Johnson Marjorie , Tabbara Nada , Galil Khadry
Published: June 2021

Worldwide research has indicated that 50‐90% of the population suffers from gingivitis, a mild form periodontal disease. Recent studies suggest that periodontitis may be detrimental to oral health and have systemic consequences, such as diabetes and stroke. Recent evidence has shown that patients value computer‐based patient education. The creation of a computer‐based tool could be highly effective in increasing dental knowledge, promoting proper oral hygiene and possibly decreasing periodontal disease rates. An online computer‐based patient education tool was created using Articulate Storyline software with assistance from the Informational Technology Resource Center at Western University. The online tool consists of 4 modules: what is periodontal disease, types of periodontal disease, causes of periodontal disease and prevention of periodontal disease. Module quizzes and a risk assessment section were also developed. Each module consists of relevant information in basic nonscientific terms, pictures and a quiz for information retention. A risk assessment section is then presented at the end in order for patients to assess their risk for developing gum disease. Future work will consider a pilot study to assess patient satisfaction within an outreach dental program.
Illustrations, CT scans and Pictures of Anatomical Dissections in a 3D Atlas of the Human Hand
Authors: Burchianti Livia , Barros Mirna , Curcio Daniella
Published: June 2021

Tridimensional anatomical atlases and apps have gained grounds in health allied educational settings and in schools for visual and fine arts. The numerous possibilities to explore different views of the studied object and observe position relations between underlying structures have helped students building more complete anatomical concepts and better understanding of neuromuscular mechanisms of the human body. We here report partial results from a project that has grown from a previous bi‐dimensional model and is now aiming at creating a 3D Atlas of the human hand. Developed by undergraduate medical students using the free 3D modeling software AutoDesk 123D Catch, the atlas is comprised by art illustrations, pictures of dissected specimens and CT scans. it allow users to enhance, hide or modify the opacity level of selected structures, so to observe anatomical features and position relations from different aspects, in superficial and deep planes. The atlas also provides essential information and quizzes involving identification and function of muscles, nerves, ligaments and bones. Art, anatomy and imaging technology are brought together to allow for a better understanding of normal function and we anticipate that the atlas may also help users to better comprehend specific conditions affecting the mechanics and/or neuromuscular control of the human hand.

Grant Funding Source:

Sao Paulo Research Foundation – FAPESP
Introducing cutting edge technology to the anatomy curriculum at the University of Pecs
Authors: Farkas Jozsef , Reglodi Dora , Nagy Gyorgy , Vigh Sandor
Published: June 2021

Modern technology is a part of our life. Today, medical doctors utilize cutting edge technology in all fields for the benefit of their patients. It is essential for medical students to learn to live and work with modern technology as early as possible in their carrier.

Turning towards the latest technology in education is a clear tendency now. Newly founded and dynamically developing universities, such as Ross University, School of Medicine, are in the vanguard of utilizing technology in education. This process is significantly slower and more difficult in old medical schools where there are strong traditions of teaching. The University of Pecs (UP), Hungary is one of Europe's oldest universities (founded in 1367 A.D.). In the Medical School at UP the leadership realized the necessity of introducing modern technology to the curriculum, while keeping and respecting the old traditions. The anatomy curriculum provides an excellent opportunity for this. In the anatomy department students meet their first “patient”. In the dissection rooms they meet the human body in the traditional way while dissecting cadavers, but at the same time an excellent opportunity is presented to introduce the latest technological innovations utilized both in medical education and clinical practice. With the support of the UP, School of Medicine, the Department of Anatomy started a robust modernization to help the future doctors to meet the challenges of the 21st century.
Fine‐tune Regulation of Cpn1 (Carboxypeptidase N1) to Control Vascular Patterning during Zebrafish Development
Authors: Wu Ting‐Yun , Wu Chang‐Yi
Published: June 2021

Vascular development is important for vertebrates and regulated by regulators via complicated signaling pathways. In our previous study, transcription factor islet2 regulated vein and ISV formation mediated by notch signaling. Genome‐wide transcriptional analysis showed that cpn1 is regulated by islet2/coupTFIb, suggested its function in vasculature. However, no direct evidence shown the role of Cpn1 in vascular function so far. Thus, we hypothesize that cpn1 encode enzymatic active subunits to perform CPN activity and function in vascular development

In‐situ hybridization showed cpn1 is expressed in developing vessels. Overexpression of cpn1 impairs the growth of ISV (intersegmental vessel) and CVP (cardinal vein plexus), which is similar to the phenotype of isl2 morphant. Knockdown of cpn1 by morpholino injection also causes vascular defects, suggesting the role of cpn1 in controlling ISV and CVP growth. We further showed the cpn1 MO knockdown works efficiently by probing cpn1 expression products and knockdown specificity by using 2nd MO. We found that a loss of cpn1 results in a decreased expression of vascular markers flk, stabilin, ephrinb2. This indicates the regulatory role of cpn1 in controlling vascular development. Promoter analysis showed cpn1 is likely regulated by isl2/coupTFIb. To examine the interaction between cpn1 and isl2/coupTFIb, we performed luciferase assay and showed cpn1 is regulated by isl2, which is consistent with our microarray results. We also revealed cpn1 is regulated by notch and VEGF signalings. Together, we showed cpn1 plays an important role for vascular development in zebrafish. We also uncovered a fine‐tune regulation of cpn1 that controls vascular patterning mediated by isl2‐Notch signaling.
Reduced Myocardial Apoptosis May Be Responsible for Attenuated Decline in Left Ventricular Systolic Function in Female Compared to Male Postinfarcted Middle‐Aged Rats
Authors: Pavliak Kristina , Chen Yue‐Feng , Pingitore Alessandro , Dedkov Eduard
Published: June 2021

Our previous study on post‐MI male and female middle‐aged rats has revealed that although both sexes had a similar scale of LV remodeling, the females showed a greater density of cardiac myocytes in the epimyocardium. However, the mechanism of this structural alteration, as well as its impact on LV function, remained unclear. Our current study was aimed at determining whether the sex‐specific difference in the density of cardiac myocytes is related to the rate of their death and whether it affects the cardiac performance in post‐MI rats. A large MI was induced in 12‐month‐old male (M‐MI) and female (F‐MI) Sprague‐Dawley rats by ligation of the left coronary artery. The cardiac function was monitored for two months post‐MI. At the end of the experiment, various LV parameters were compared between sexes. We found that although both groups had a similar pattern of decline in LV function, the F‐MI rats exhibited a less deteriorated cardiac performance compared to M‐MI rats, as indicated by the values of stroke volume (0.18±0.01 vs. 0.14±0.02 mL, P<0.05), ejection fraction (39.9±3.4 vs. 26.7±7.7 %, P<0.05) and cardiac output (57.8±3.1 vs. 44.6±8.7 mL/min, P<0.05). Moreover, the F‐MI rats displayed a higher density of cardiac myocytes in the epimyocardium than the males (2383.8±242.6 vs. 1785.7±55.9 cells/mm2, P<0.05). This was associated with a lower rate of myocyte apoptosis in the F‐MI rats (0.18±0.08 vs. 0.91±0.30 apoptotic cells/mm2, P<0.01). Thus, our data showed that the F‐MI rats had a better preserved LV function compared to M‐MI rats, and that this might be caused by an increased myocyte survival in females.
Effects of Bleomycin, Etoposide and Cisplatin Treatment on Leydig Cell Structure and Transcription of Steroidogenic Enzymes in Rat Testis
Authors: Kilarkaje Narayana , Al‐Bader Maie
Published: June 2021

Anticancer chemotherapy affects pituitary‐testicular hormonal axis in humans and animals. This study investigated the effects on Leydig cells of three cycles of bleomycin, etoposide and cisplatin (0.75, 7.5, and 1.5 mg/kg, respectively; BEP) chemotherapy in rat testis. The chemotherapy has induced hyperplasia of and degenerative changes in Leydig cells at the end of BEP exposure, which remained so even after a recovery time of 63 days. The increased testicular oxidative stress at the end of the chemotherapy returned to the normal level after the recovery time. The chemotherapy has stimulated the transcription of scavenger receptor class type‐B1 (SCARB1), steroidogenic acute‐regulatory protein (StAR), cytochrome P450 cholesterol side‐chain cleavage (CYP11A1), CYP17A1, and inhibited that of 17β‐hydroxysteroid dehydrogenase (HSD17B6) and CYP19A1 in association with increased cholesterol and decreased testosterone levels. After the recovery time, the chemotherapy still had inhibitory effects on the transcription of all the above genes and luteinizing hormone receptor and HSD3B1, but not on the StAR gene. The cholesterol and testosterone levels also did not show any significant changes. The decreased testosterone level at the end of the chemotherapy was probably due to inhibition of HSD3B1 and HSD17B6 genes. In conclusion, the BEP chemotherapy induces Leydig cell hyperplasia and alters testicular steroidogenic pathway in rats (Supported by Kuwait University Grant # MA02/08 and SRUL02/13).
Get up and Learn – Active Learning of Hemoglobin Function in the Biochemistry Classroom
Authors: Diegelman‐Parente Amy
Published: June 2021

Constructivist learning theory, as formalized by Piaget, suggests that learners construct new knowledge from their experiences and is best facilitated with pedagogic approaches that promote learning by doing. A traditional stumbling block in undergraduate biochemistry is the structure‐function relationship in proteins. Students often first encounter this relationship in the study of the oxygen‐binding proteins hemoglobin and myoglobin. While the molecular structures and physiological roles of these proteins are known in detail, the interplay of additional components (e.g. carbonic anhydrase, O2/CO2 levels, 2,3‐BPG) can make this dance in the circulatory system a challenge for students to master conceptually. In this study, an active learning approach through a hands‐on game was used to facilitate student mastery, whereby students become the molecules responsible for oxygen transport in the circulatory system. By students having a greater understanding of how this system works properly in a healthy state, they can better understand aspects that result in disease (e.g. Sickle‐cell anemia and hemoglobin variants) or predict outcomes arising from changes in environmental conditions (e.g. high altitude or hyperventilation). The objective of this study is to determine if there is any difference between perceived and actual success in learning outcomes as students partake in this alternative pedagogical approach. The working hypothesis is that utilization of constructivist pedagogies, such as an active learning game, will result in higher perceived and actual student success. Data is being collected during the Fall 2015 semester to evaluate this hypothesis.
Examining microbial biodiversity in soil: A research‐based introductory biology laboratory course
Authors: Lo Stanley
Published: June 2021

We report a novel introductory biology laboratory course designed as an independent, on‐going research program, with students as the primary researchers. This course aims to develop an understanding for biology through inquiry‐based laboratory experiments, and specific learning objectives are listed in Table 1. Students work in teams to collect, analyze, and present original data while learning core concepts and laboratory methods. Specifically, we examine if and how soil microbiomes are different for native vs. invasive plant species in the Southern California diversity hotspot. Students analyze soil properties such as moisture and pH and correlate them to functional and genetic biodiversity of microbiomes, assessed by carbon source utilization and 16S rDNA sequencing. Together, these elements span macroscopic to molecular scales, providing opportunities to connect student learning across biology disciplines.

This presentation will focus on the design, implementation, and evaluation of the course. Laboratory experiments are designed to be completed by introductory students within a 3‐hour laboratory and 1.5‐hour lecture per week while allowing for the reliable collection of research data. We found that student‐generated data are consistent across groups: soil genomic DNA preparations and PCR clean‐up reactions have standard deviations <10% of average DNA concentrations, and >95% of PCRs result in products at appropriate sizes. Student outcomes are evaluated by pre‐ and post‐surveys and ‐tests to track changes affective domain (Science Motivation Questionnaire II), self‐efficacy in research skills (Classroom Undergraduate Research Experience survey), and ability to draw conclusions based on data (open‐ended questions that test scientific argumentation).

Learning objectives
Activities
Student assessments

Understand functional and genetic biodiversity
Discuss material in laboratory and lectures
Quizzes in lectures, laboratory reports

Develop testable hypotheses and design controlled experiments
Carry out scientific processes in the context of the course
Data collection by students, laboratory reports

Draw conclusions based on data and evidence
Analyze and discuss laboratory data using statistics
Figures and results in laboratory reports

Communicite scientific results in a publication‐style laboratory report
Write, review, and revise laboratory reports
Laboratory reports, peer review of laboratory reports
Implementation and Benefits of a Capstone Undergraduate Research Program
Authors: Martin Debra
Published: June 2021

The benefits of an undergraduate research opportunity have been well documented. At Saint Mary's University of Minnesota, undergraduate research in the Biology and Chemistry Departments has been an integral requirement of the majors for nearly 70 years. The requirements are that the students complete an original research project that must be laboratory or field‐oriented. Courses in research design and implementation of the research prepare students to perform and accomplish the capstone research project. Students are assessed on their ability to communicate by utilizing both a written thesis and oral presentation. This poster will outline the learning goals, learning outcomes, and assessment tools used to monitor student progress as they gain confidence to become an independent researcher.
De novo Thymidylate Biosynthesis and Uracil Misincorporation in DNA as Related to Neural Tube Defect and Cancer Outcomes in Mouse Models
Authors: Field Martha , Lan Xu , Stover Patrick
Published: June 2021

In the absence of adequate thymidylate (dTMP) synthesis, uracil is misincorporated into DNA during replication and/or repair, as DNA polymerases do not distinguish dTTP from dUTP. Current dogma holds that the dUTP/dTTP ratio determines the rate at which dU is incorporated into DNA. Here we show that the dTMP precursors uridine (U) and deoxyuridine (dU) have distinct fates in DNA. Radioactive tracer studies in HeLa and mouse embryonic fibroblast (MEF) cells demonstrate that exogenous dU is incorporated into DNA as the base dT, and not misincorporated as dU. Exogenous U is incorporated primarily as the pyrmidine base dC. In HeLa cells both exogenous U and dU modestly stimulate proliferation, but U in culture media also increase caspase 3/7 activity, a marker of apoptosis. Exogenous U and dU also have distinct and opposing effects on cytotoxicity of 5‐flourouracil (5‐FU) in HeLa cells.

The ApcMin/+ and Shmt1‐/‐ mouse models were used to investigate whether supplementation of folate‐deficient diets with U, dU, or thymidine (T) affected colorectal cancer or NTD incidence, respectively. Unexpectedly, exencephaly in offspring from Shmt1‐/‐ mice was rescued by dietary dU but exacerbated by dietary U. Conversely, U supplementation reduced intestinal tumor number in ApcMin/+ mice by more than 30% relative to the unsupplemented diet, whereas dietary dU did not affect tumor number. Supplemental thymidine did not affect the phenotype of either mouse model. Taken together, these data challenge the assumption that uracil accumulation in DNA is driven simply by the dUMP/dTMP ratio.
High Fat Diet‐Induced Alterations in Alternative Splicing of Pre‐mRNA
Authors: Ravi Suhana , Schilder Rudolf , Jefferson Leonard , Kimball Scot
Published: June 2021

Previous studies have shown that the pattern of alternative splicing of the fast troponin T (Tnnt3) pre‐mRNA in gastrocnemius muscle is directly proportional to increases in both experienced body mass (i.e. actual body weight plus additional external load) and muscle force production. However, this proportionality is disrupted in obese Zucker rats, such that the muscle of obese rats expresses Tnnt3 splice forms typically observed in smaller animals. The objective of the present study was to assess the time course of development of the obesity‐induced impairment of Tnnt3 pre‐mRNA alternative splicing as a prelude to delineating the molecular mechanism(s) involved in the response. In this study, obesity‐prone Sprague Dawley rats were placed on either a control or a high‐fat diet (HFD) for up to 8 wk. There was no significant difference in body weight in rats fed the HFD compared to the control diet. However, the pattern of Tnnt3 pre‐mRNA alternative splicing was altered after only 1 wk of consuming the HFD, such that there was a mismatch between body weight and the relative abundance of Tnnt3 splice forms. Moreover, the mismatch in alternative splicing worsened with increasing time on the HFD. For example, the relative abundance of Tnnt3 pre‐mRNA present in α‐splice forms increased with increasing body weight in rats fed the control but not the HFD. Furthermore, exon array analysis identified a significant number of other pre‐mRNAs that were alternatively spliced in the gastrocnemius in response to consuming the HFD for 8 wk. Overall, the results indicate that early changes in alternative splicing associated with feeding a HFD occur prior to changes in body weight and thus are likely a direct effect of the diet. (supported by NIH grant DK094141)