bluuebunny/crossref_metadata_2025_split huggingface dataset:
Augmenting Medical Neuroanatomy Cadaveric Dissection with DiceCT Imaging
Authors: Bertolino Blake , Speer Weston , Gignac Paul , O’Brien Haley 
Published: April 2020

Dissection of the brain and central nervous system for medical education is an informative but often wholly destructive process, leaving students with little reference material from which to review neural structures and their anatomical relationships. Here, we explore the use of diffusible iodine contrast enhancement followed by high‐resolution microCT (diceCT) to combine destructive physical dissection with more permanent digital dissection. DiceCT methods render soft tissues radiodense to X‐rays, emulating the tissue discrimination of magnetic resonance imaging (MRI). Such imaging studies provide students the opportunity to identify and interpret pathological findings in ex‐vivo neural tissues. To identify best‐practices for incorporating diceCT imaging into the medical neuroanatomy lab, we stained formalin‐preserved human brains with three concentrations of iodine (3, 4, and 5% aqueous I2KI) for 3 weeks, replacing the iodine solutions after the first and second weeks. All brains were microCT scanned at 63‐micron resolution, and the resulting image stack was rendered in Avizo version 2019.2. The stained brains were then bisected, and one hemisphere was de‐stained for 4 weeks by alternating baths of 1% sodium thiosulfate with deionized water each on a week‐long cycle. All concentrations of iodine were effectively de‐stained using this protocol. Students then performed comparative physical dissections on never‐stained, stained only, and stained‐then‐de‐stained brains, and comparative digital dissections were performed on brains of each iodine concentration. In physical dissection, stained only brains had few advantages—although the dark iodine stain enhanced some fiber tracts, specimens were smaller, physically difficult to manipulate, and harbored subcortical nuclei (e.g. the basal ganglia) that were more difficult to identify. Dissection of never‐stained and de‐stained brains was found to be equivocal, with de‐stained brains regaining much of their original size. In digital dissection, the 5% aqueous I2KI solution provided the clearest discrimination between structures, and pathologies such as age‐related atrophy and hydrocephalus were apparent. In addition to leaving a more permanent imaging study record of nervous tissue specimens, digital dissection allows for 3D printing of brain regions. These benefits were positively received by students, and we recommend incorporation of rapid, inexpensive tissue staining and tomography to enhance participation and clinical skills in the anatomy lab.Support or Funding InformationFederal Work Study, OSU Center for Health Sciences
Claudin‐1 responsiveness through modulation of the inflammatory status in the Caco‐2 model epithelium.
Authors: Largoza Gabrielle E. , Carr Nicholas R. , King Jonathan M. 
Published: April 2020

The use of hydrocortisone in association with indomethacin is a known risk factor for spontaneous intestinal perforation (SIP) in extremely low birthweight infants. While hypoxia, inflammation, and hydrocortisone have been found to independently alter both tight junction functionality and formation, the concomitant effects of inflammation, hydrocortisone, and indomethacin have yet to be well defined. Through the use of the Caco‐2‐BBe cell line as an in vitro model of the intestinal epithelial barrier, we tested the effects of the pro‐inflammatory agents TNF and IFNγ and the anti‐inflammatory agents indomethacin and hydrocortisone on barrier integrity and tight junction protein expression. Confocal imaging and cellular ATP analysis after 24 and 48 hours of exposure to the inflammatory agents both showed no significant changes in cellular proliferation or ATP synthesis, respectively. Western blot analysis found that claudin‐1 protein expression and localization is affected by inflammatory cytokines. Live cell confocal imaging of eGFP‐claudin‐1 Caco‐2 cells found that addition of pro‐inflammatory agents or the addition of anti‐inflammatory agents resulted in rapid dynamic junctional reorganization. When both the pro‐ and anti‐inflammatory agents were added, this pattern of reorganization was also seen. However, the concomitant effects of the combined pro‐ and anti‐inflammatory agents resulted in a profound inability to recapitulate its original claudin‐1 configuration. The lack of an effect on cellular proliferation and ATP synthesis paired with the alterations in claudin‐1 protein levels and localization suggests that claudin‐1 may be a potential target in better understanding the progression of spontaneous intestinal perforation.Competing Interest StatementAll authors have indicated they have no potential conflicts of interest to disclose. The view(s) expressed herein are those of the author(s) and do not reflect the official policy or position of Brooke Army Medical Center, the U.S. Army Medical Department, the U.S. Army Office of the Surgeon General, the Department of the Air Force and Army, Department of Defense or the U.S. Government.Support or Funding InformationTrinity University Biology Department
Extrahepatic Biliary Tract Anatomy: A Review
Authors: Giron Felipe Manuel , Ramírez-Buchheim Daniela Paola , Pizano Alejandro Manuel , Rueda-Esteban Roberto Javier 
Published: April 2020

Laparoscopic cholecystectomy is one of the most common surgical procedures in general surgery. Significant gaps in knowledge still exist and even with clinical expertise, there might be limitations. Some of these are exploration restrictions during laparoscopic approach, visualization impairments and distortion of the usual arrangement of structures because of local inflammation, among others.Given the wide variability in anatomical presentation and considering that even in 56% of the population there is a close relationship to main vascular structures, intraoperative injuries might occur leading to higher rates of complications, longer surgical time and increased morbidity.Our aim is to describe the most common and relevant anatomic variations in the extrahepatic biliary tract (EBT). A systematic review of the literature was pursued using the MeSH terms: “Biliary Tract/abnormalities”, “Bile Ducts and Extrahepatic/abnormalities”. Only articles written in Spanish and English and limited to the last 20 years were considered.The anatomical structure considered normal for the EBT is described in roughly 60% of the population. The most variable structures according to included manuscripts are the left and right biliary ducts and the most prevalent variations are: posterior branch of the right hepatic duct draining into the left hepatic duct, triple confluence of the intrahepatic ducts, and anterior or posterior branch of the right hepatic duct draining into the common hepatic or cystic duct.As mentioned before the awareness and comprehension of EBT variations is crucial in avoiding complication, morbidity and achieving optimal outcomes in diagnostics and therapeutic procedures as reducing intraoperative iatrogenic injuries and considering the need of noninvasive techniques for EBT identification for a better surgical planning.
Sex‐dependent Liver Volume Recovery and Hemodynamics Measured by Non‐Invasive Ultrasound Imaging after Partial Hepatectomy in Rats
Authors: Barnhart Benjamin , Kan Toshiki , Wessner Corinne E. , Eisenbrey John R. , Hoek Jan B. , Vadigepalli Rajanikanth 
Published: April 2020

Initiation of liver regrowth after partial hepatectomy (PHx) is often linked to increased flow per unit tissue through the hepatic portal vein. Previous studies of liver hemodynamics have used surgical implants, however ultrasound‐based methods can non‐invasively measure blood flow, organ volume, and other physiological parameters.Male and female Sprague‐Dawley rats were pair‐fed liquid diet containing ethanol or carbohydrate per Lieber DeCarli protocol for 5–6 weeks prior to a 70% PHx in which left lateral and medial lobes were surgically removed. A VEVO 2100 (VisualSonics, Toronto, Canada) high‐frequency ultrasound system with a motorized‐step ultrasound transducer was utilized to calculate liver volume using a 3D mapping software. Shear‐wave elastography measured liver stiffness with an Aplio i800 ultrasound system (Canon Medical Systems, Tustin, CA, USA). Color and Spectral Doppler evaluated blood flow velocity in the portal vein (PV) and the main hepatic artery (HA) at their entry into the right lobe. Laser coupled photoacoustic imaging aided in quantification of oxygen saturation in the liver tissue.Liver regrowth as measured by liver volume was impaired in ethanol‐fed male rats as early as 48 hours after PHx, only recovering ~70% of initial volume by one week. In contrast, all other animals, including ethanol‐fed females, had recovered to ~100% of liver volume by one week post‐PHx. All animals showed increased PV and HA blood flow per liver volume immediately after PHx. Relative PV flow continued to increase by the 12‐hour time point, while relative HA flow had variable changes. Ethanol‐fed males had decreased ratio of PV/HA flow compared to all other animals prior to PHx. In the first 24 hours after PHx, relative HA flow decreased or remained stable for most groups, but increased for ethanol‐fed females. Liver stiffness increased immediately after PHx and again by 10 hours after PHx for all groups, though never reaching values that would suggest fibrosis. Stiffness remained elevated for at least 96 hours for all groups, and resolved by one week post‐PHx. Photoacoustic imaging showed liver tissue oxygen saturation fluctuating between 50–60% for all animals, with no discernable patterns.Ethanol‐fed male rats had an impaired liver volume recovery response after PHx, however this ethanol‐mediated impairment was not seen in females. Interestingly, ethanol‐fed males were the only group to have a decreased PV/HA flow ratio prior to PHx, while ethanol‐fed females were unique in their prolonged increase in HA flow at 10 hours post‐PHx. Taken together, these results implicate increased HA flow as a compensatory mechanism in chronic ethanol‐fed rats.Support or Funding InformationSupported by NIH NIAAA R01 AA018873, T32 AA007463 and NIBIB U01 EB023224.
Drug Discovery for Multiple Hereditary Exostoses
Authors: Weiss Ryan , Rother Sandra , Griglio Alessia , Oukoloff Killian , Heynen-Genel Susanne , Ballatore Carlo , Esko Jeffrey 
Published: April 2020

Multiple hereditary exostoses (MHE) is an autosomal dominant disorder that results from heterozygous loss‐of‐function genetic mutations in exostosin 1 (EXT1) or exostosin 2 (EXT2). One in 50,000 children worldwide are affected by this relatively rare pediatric disease, which is characterized by osteochondromas that form next to the growth plates of skeletal elements. EXT1 and EXT2 encode the copolymerase responsible for biosynthesis of the backbone of heparan sulfate (HS), a cell surface polysaccharide. Heterozygous mutations in EXT1 or EXT2 lead to shortened HS chains in plasma and various tissues. Currently, there are no approved therapies for this disease; therefore, there is much interest in preventing and/or slowing the growth of exostoses through the development of small molecules that enhance HS levels on the cell surface. In this study, a high‐throughput phenotypic screen was developed to discover potential small molecule drug candidates that augment HS expression in mammalian cells. Screening of a compound library of ~57,000 molecules resulted in the discovery of several N‐aryl‐2‐aminothiazoles that displayed favorable “drug‐like” physicochemical properties, exhibited minimal cytotoxicity, and enhanced HS expression up to 3‐fold on the cell surface. Preliminary studies of structure‐activity relationships identified active analogs of the primary hit and key physicochemical properties required for their biological activity. Ongoing studies aim to expand the SAR studies, identification of a preferred candidate for in vivo studies, and finding the biological target for these compounds. Overall, these studies provide a promising new avenue for the development of novel therapies for MHE and other disorders involving glycosaminoglycan deficiencies.Support or Funding InformationR01AR064202‐03 (to J.E.), K12HL141956 (to R.W.)
Establishment of screening for agents for improving dysgeusia using medaka
Authors: Tsutsumi Rie , Beppu Kana , Morio Atsushi , Sakaue Hiroshi , Tsutsumi Yasuo M. 
Published: April 2020

ObjectsDysgeusia are caused by various factors such as chemotherapy, drugs, infection, and aging, and many people suffer. However, effective treatments for dysgeusia have not been established for drugs and foods. We have previously reported that the expression of lingual taste receptor T1R3 is reduced during chemotherapy induced dysgeusia. On the other hand, medaka, one kind of small fish, has many taste buds from mouth to gill, and the number is 5–6 times that of mice. In this study, we created a medaka that expressed the T1R3 gene with a fluorescent tag, and tried to visualize dysgeusia.MethodsWe established T1R3 knock‐in medaka using Crisper/cas9 system. The construct was microinjected into eggs immediately after medaka laying, and knock‐in was confirmed with a fluorescence microscope. It was confirmed that expression of T1R3 disappeared when reared in a solution containing an anticancer drug (5FU or platinum agent) using medaka of the F1 generation or later.ResultsWe succeeded in producing a medaka that visualized the taste receptors by microinjecting fluorescently labeled T1R3 into medaka eggs. The expression of T1R3 disappeared when reared in a solution containing an anticancer drug whereas several amino acids such as glutamate and overeating were effective in maintaining and increasing T1R3 expression.ConclusionsWe established screening for agents for improving dysgeusia using medaka.Support or Funding InformationThis study was supported by Japan Society of Promotion Science #18K11075 to RT.
Factors contributing to the equal forelimb and hindlimb stride lengths during quadrupedal walking
Authors: Johnson Laura E. , Zeininger Angel , Schmitt Daniel 
Published: April 2020

The forward motion of terrestrial vertebrates can be divided into stride lengths (distance traveled by the body from touchdown to the next touchdown of a limb) which is the sum of the lengths of stance and swing phases for each limb. Those lengths are influenced by the anatomy of the limbs; longer limbs, for example, produce longer steps and strides. During typical quadrupedal walking, at least one forelimb and hindlimb are in contact with the ground at any given time. This means the fore‐ and hindlimbs (regardless of limb proportions) should have the same stride length, though the length of the phases (stance, swing) may differ, as long as they sum to equal stride lengths. In addition to the anatomical limb length (ALL), joint excursion patterns, which often differs between the forelimb and hindlimb can influence stance and step. Animals can behaviorally adjust their limb postures to change their effective limb lengths (ELL, the minimum distance between the shoulder/hip and wrist/ankle) in order to achieve the necessary stride length. Thus, we hypothesize that forelimb and hindlimb stride lengths should be equal, but this may be achieved in different ways and therefore all other parameters may vary possibly in relation to limb proportions.To study this, we examined 69 quadrupedal walking bouts of four New World monkeys with differing ALL. Preliminary analyses indicate a close correlation of forelimb and hindlimb stride length (0.93, p<0.01), regardless of differences in ALL proportions. Factors that contribute to an equal stride length are more variable. Forelimb and hindlimb stance phase length were highly correlated (0.700, p<0.01), and forelimb and hindlimb swing phase length were highly correlated (0.834, p<0.01). Although significant, a low correlation of the length of swing and stance phase was found for both the forelimb (0.412, p<0.01) and the hindlimb (0.430, p<0.01). When modeling step length in stance by angular excursion and ELL, the model accounts for an average of 85% of the distance traveled. Taken as a whole, these data, especially the intralimb and interlimb patterns, suggest that the step and swing lengths are not easily predicted by limb anatomy or effective limb lengths alone. Thus, other factors must also contribute to step length and thus equal stride lengths, such as bending of the back and contributions made by hand and foot posture. Overall, these data indicate that although stride lengths do need to be equal between the forelimb and hindlimb, contributing parameters vary between strides and may vary by species anatomy and ecology.Support or Funding InformationNSF BCS‐9222526
Re‐designing Biochemistry Laboratory to Implement BASIL CURE: from Structure to Function, from Student to Researcher
Authors: Goodman Anya L. , Jones Eric M. , Laubscher Andrea M. , Henry Jack , Craig Paul A. 
Published: April 2020

Course‐based undergraduate research experiences (CUREs) have recently emerged as an alternative to traditional science laboratory instruction. Cross‐institutional faculty communities help crowd‐source curricular design and assessment, as well as facilitate data sharing and trouble‐shooting. As participants in the Biochemistry Authentic Scientific Inquiry Laboratory (BASIL) community, we re‐designed our upper level biochemistry laboratory course to incorporate CURE. Scientific objectives of BASIL focus on characterizing biochemical activities of proteins with known structure and unknown function. BASIL curriculum (freely available on GitHub
https://basilbiochem.github.io/basil/) includes modules on computational predictions of protein function, protein purification, and analysis of biochemical activity and kinetics.Over the course of four years, our implementation has evolved to accommodate local needs and constraints and to address technical, logistical and pedagogical challenges we encountered along the way. We discuss this evolution in light of recommendations for CURE course design (Kloser et al., 2011) and the five dimensions of CURE (scientific practices, discovery, relevance, collaboration, and iteration; Auchincloss et al., 2014). Three main challenges for our implementation included 1) scaling of research training from the “apprentice” model to the classroom, 2) alignment of course learning objectives, activities and assessments with research goals to create optimal conditions for productive struggle, 3) student buy‐in.CURE implementation presents challenges with timing of research experiments in a scheduled course, availability of equipment, management of reagent requests and hazardous waste. Instructional support staff (AML) was critical for overcoming a number of logistical, technical and pedagogical challenges. In addition, undergraduate students pursuing research projects outside of class contributed to the CURE by developing new protocols, testing new proteins or generating computational predictions using tools that were outside the scope of the course. To understand if/how students in our course develop laboratory research skills, we began assessing anticipated learning outcomes previously defined by Irby et al. (2018), focusing on students’ evaluation of protein purification experiments. Initial assessment results revealed learning difficulties and led to modifications in instructional activities, including use of simulated experiments. To gauge student frustration and to help students improve their metacognitive skills, we asked students to reflect on their experiences in short weekly structured journal entries. Students’ negative perceptions of failed experiments, potentially, rooted in fixed mindset, require additional intervention.Support or Funding InformationSupported by NSF‐IUSE #1503676 to ALG, #1710538 to ALG and EMJ; 1503811 and 1709170 PAC.
To bee inside or outside? Impacts of overwintering environment on gene expression of the alfalfa leafcutting bee, Megachile rotundata
Authors: Cambron Liz D. , Yocum George D. , Greenlee Kendra J. 
Published: April 2020

Diapause is a non‐feeding stage that many insects go through to survive the winter months. How nutrition is regulated during these months of overwintering is still unclear. Diapausing insects are exposed to environmental stressors including temperature stress, but responding can be energetically costly. With fixed resources, overall metabolism and insulin signaling are maintained at low levels, but it is unclear if those pathways change in response to seasonal temperature fluctuations. Our hypothesis is that insulin signaling is responsible for allocating energy in response to fluctuations in temperature during overwintering months. To test this hypothesis, we overwintered alfalfa leafcutting bees, Megachile rotundata, in either a lab setting at a constant 4°C or in the field in naturally fluctuating temperatures, collected samples monthly throughout the winter, and then measured expression of genes in the insulin pathway using nCounter analysis (NanoString Technologies Inc.). nCounter is a highly multiplexed single molecule counting technique. A Dunn’s test was performed to analyze the effects of time and location for all possible comparisons. Our results showed that several genes were differentially expressed over time and between locations. However, more changes were seen in field samples, possibly in response to environmental cues. With a hierarchical cluster, field samples showed three distinct month clades, possibly correlating with different developmental stages: diapause, post‐diapause quiescence, and direct development. Although several genes were differentially expressed, many of them were downstream of the insulin pathway (Foxo, MAPK, GSK3β). This could mean that components of the insulin pathway are being utilized to respond to temperature fluctuations in the field. Along with evidence from other studies, our results suggest that insects may use different mechanisms for diapause regulation in response to their environment. This plasticity in regulation may be achieved by using targets of other pathways such as that of the insulin pathway. There is no denying that pollinators are crucial for the health of our ecosystem, but before we can save the bees, we need a better understanding of how their overall physiology is impacted in a drastically changing environment.Support or Funding InformationThis research was funded, in part, by grants from the National Science Foundation Graduate Research Fellowship and an NSF Graduate Research Internship with the USDA‐ARS Fargo, ND, and NSF EPSCoR‐1826834 to KJG.