bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effects of eating a high fat or a ketogenic diet on sensitivity of female rats to methamphetamine
Authors: Elsey Madeline K , Beltran Nina M , Serafine Katherine M 
Published: April 2020

Eating a high fat diet leads to negative health consequences such as obesity, type 2 diabetes, and dysfunction to dopamine systems. For example, eating a high fat laboratory chow enhances the sensitivity of rats to the behavioral effects of drugs of abuse (i.e., methamphetamine). Previous research has focused on diets that are high in fat and carbohydrates but low in protein. In contrast, a ketogenic diet is high in fat, low in carbohydrates, and low in protein. Ketogenic diets have been used in humans for the treatment of epilepsy and have been investigated for weight loss. Preclinical studies suggest that rats eating a ketogenic laboratory chow are less sensitive to the locomotor effects of cocaine (e.g., stereotypy). Based on this previous literature, it was hypothesized that eating high fat chow would enhance sensitivity of rats to methamphetamine‐induced locomotion and sensitization; however, rats eating ketogenetic chow would not differ from rats eating standard chow. To test this hypothesis rats eating standard chow (17% kcal from fat, 58% kcal from carbohydrate, 25% kcal from protein), high fat chow (60% kcal from fat, 21% kcal from carbohydrate, 18% kcal from protein), or ketogenic chow (90.5% kcal from fat, 0.3% kcal from carbohydrate, 9.2% kcal from protein) were tested once weekly with cumulative doses of methamphetamine (0.1–3.2 mg/kg; i.p.). After 4 weeks, rats eating high fat chow were more sensitive to the locomotor‐stimulating effects of smaller and intermediate doses of methamphetamine (e.g., 0.32, 1.0 mg/kg) than rats eating standard chow. Rats eating ketogenic chow were also more sensitive than rats eating standard chow to the locomotor‐stimulating effects of methamphetamine, but only at the largest dose tested (3.2 mg/kg). Future research will investigate the mechanisms underlying the effects of diet on sensitivity of rats to methamphetamine‐induced locomotion and sensitization, as well as examine the impact of these and other diets on sensitivity of rats to the rewarding and reinforcing effects of drugs of abuse.Support or Funding InformationThe authors have no conflicts to disclose. This work was supported by the National Institutes of Health, through the National Institute of General Medical Sciences under linked award numbers RL5GM118969, TL4GM118971, and UL1GM118970 and award number R25GM069621. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Elucidating the Mechanisms of Developmental Cell Death in the Retina Using Stem Cell‐derived Organoids
Authors: Brooks Tara , Park Yuna K. , Vielle Anne , Vergara Natalia 
Published: April 2020

The development of stem cell technologies has opened up new opportunities for research and therapeutic development that, until recently, were unavailable. In particular, the development of human stem cell derived 3D tissues or “organoids” that recapitulate the native histoarchitecture, and to some degree the native functionality, have provided incredible advancements in our understanding of human physiology and disease. In this study we take advantage of retinal organoids as a new model to study the mechanisms of cell death affecting retinal ganglion cells during human development.Human induced pluripotent stem cells (hiPSC) were directed to follow a neuronal lineage and were then further differentiated into 3D retinal tissue. Retinal organoids were collected at various time points during development and analyzed using Western blot and immunofluorescent staining for retinal ganglion cells and programmed cell death markers. Our results show the role of programmed cell death as a mechanism to control retinal ganglion cell numbers during development. This has important consequences for understanding not only basic developmental processes, but also the basis of congenital retinal abnormalities that can lead to diseases of vision. Moreover, this knowledge has potential impact for translational research using retinal organoid models.Support or Funding InformationThis work was supported, in part, by a Challenge Grant to the Department of Ophthalmology at the University of Colorado from Research to Prevent Blindness.
Apo‐hemoglobin and Haptoglobin Limit Acellular Hemoglobin Negative Effects
Authors: Munoz Carlos Jose , Pires Ivan S. , Buehler Paul W. , Palmer Andre F. , Cabrales Pedro 
Published: April 2020

BackgroundHaptoglobin (Hp), a scavenger protein for hemoglobin, and apo‐hemoglobin (apoHb), a chemically synthesized scavenger protein for free heme, present promise in preventing acellular hemoglobin toxicity. The purification process of both molecules is cost effective and can help a number of patients suffering from a septic shock, thalassemia, sickle cell and various other diseases.MethodsSystematic parameters, microhemodynamics and pharmacokinetics were observed on Golden Syrian hamsters (n=32) using intravital microscopy on the dorsal skin fold. Hamsters were injected with apoHb, Hp, and a combination of apoHb and Hp via the carotid artery. This was followed by a single 10% top‐load of acellular Hb via the carotid arteryResultsA combined treatment with apoHb and Hp was found to reduce acellular hemoglobin toxicities. Systematic parameters and microhemodynamics of all sized arterioles and venules resulted in sustained values when compared to baseline and other treatments (apoHb and Hp alone). The pharmacokinetics data suggests strong scavenging functionality over 6 hours.ConclusionThe results suggest a promising treatment against acellular hemoglobin and hemolytic anemia induced toxicity where the apoHb is stabilized in a molar excess of Hp preventing vasoconstriction and preserving microvascular blood velocity and blood flow. Ultimately, these results support the efficacy of using purified Hp and apoHb to control hemoglobin and heme induced toxicity.Support or Funding InformationThis work was supported by the NIH Heart Lung and Blood Institute under Grants T32‐HL105373, R01‐HL126945, and the DOD DMRDP under Grant W81XWH‐18‐1‐0059
Purification of Insoluble Protein Aggregates from Skeletal Muscle Using a Pre‐Clinical Model of Huntington’s Disease
Authors: Turner Kristen D. , Opatrny Lauren M. , Fuqua Jordan D. , Musetescu Andrei , Kronemberger Ana , Lira Vitor A. 
Published: April 2020

Insoluble protein aggregates are distinctive attributes of several tissues affected by aging and are suggested to play a pathological role in the development of neuromuscular disorders (i.e., Huntington’s Disease). However, the influence of insoluble protein aggregates in skeletal muscle on muscle‐specific function is unclear. This is due, in part, to the absence of standardized protocols for isolating protein aggregates so they can be identified and further investigated in vivo. The overall aim of the present methodological study was to determine the most effective method for isolating insoluble protein aggregates from mouse skeletal muscle. This study was conducted using a pre‐clinical Huntington’s Disease model. Electroporation with plasmids encoding EGFP fused to exon 1 of human huntingtin containing 23 (EGFP‐Q23, control) or 74 polyglutamine repeats (EGFP‐Q74, aggregate‐prone) in contralateral TAs was performed (N = 9 muscles/group). Using three adapted protocols from the literature that combined mild detergents and centrifugation to purify insoluble and soluble fractions (Protocol 1 (1% Triton‐X‐100, pH 7.5), Protocol 2 (1% NP‐40, pH 7.4) and Protocol 3 (0.5% Triton‐X‐100, pH 8.0)), we hypothesized that the most efficient protocol would show aggregate‐prone proteins (i.e., EGFP‐Q74) accompanied by adaptors involved in the degradation of protein aggregates by autophagy (p62, NBR1), enriched in insoluble fractions. Approximately 6‐, 5‐ and 3‐fold increases in EGFP‐Q74 vs. EGFP‐Q23 were observed in the insoluble fractions of Protocols 1, 2 and 3, respectively (P < 0.05). 2.5‐fold increases in adaptor p62 in the same insoluble fractions from Protocols 1 and 2 were observed, while only a 2‐fold increase was observed with Protocol 3. Lastly, we observed trends for NBR1, ubiquitinated proteins and total proteins to be elevated using Protocol 1. These observations indicate that Protocol 1 and 2 might be more effective than Protocol 3 for isolating protein aggregates from skeletal muscle and these findings should be considered in future studies extracting insoluble proteins using this Huntington’s Disease model.Support or Funding InformationThis research is supported by an SDG grant from the American Heart Association (V.A.L.).
Anoxic Turtles May Be Blind, but Not Deaf: Responses of the In Vitro Pond Turtle Brain to Visual and Auditory Stimuli During Anoxia
Authors: Ariel Michael , Warren Daniel E. 
Published: April 2020

Freshwater pond turtles are among the most anoxia‐tolerant vertebrates, an ability that allows some species to overwinter for months in ice‐locked ponds without oxygen. To determine if anoxic slider turtles can maintain the ability to detect visual and auditory stimuli, we monitored field potentials in an in vitro reduced brainstem while superfusing with either oxygenated or anoxic superfusate. Visual stimuli were recorded using glass recording pipettes placed medial to the optic tectum while flashing light on the eyes. For auditory stimuli, the pipette was placed medial to the eighth cranial nerve (the presumptive cochlear nucleus). In place of sound stimuli, a glass rod with a small polished tip was mounted to a piezo‐electric piston that moved onto the submerged tympanic membrane in brief inward and outward steps. Auditory responses of the brainstem occurred after a very brief latency to each step of the tympanic membrane, while there was a long latency to each light flash in the optic tectum
. An optical probe measured the concurrent oxygen tensions under control (oxygenated) and anoxic conditions. After switching from control to anoxic superfusate, there was no substantial change in the amplitude of the averaged auditory response to steps of the tympanic membrane. However, responses recorded in the optic tectum to light flashes while bathed in anoxic superfusate decreased strongly, but recovered upon return to the control superfusate. These results suggest that turtles submerged for long periods in anoxic ponds may retain sensory responses to auditory but not visual stimuli.Support or Funding InformationNational Science Foundation CAREER Award (1253939) to DEW.
Drosophila TRIM32 cooperates with glycolytic enzymes to promote cell growth
Authors: Bawa Simran , Brooks David S. , Neville Kathryn E. , Sagar Md Abdul Kader , Geisbrecht Brian V. , Tennessen Jason M. , Tipping Marla , Elicieri Kevin W. , Geisbrecht Erika R. 
Published: April 2020

Control of tissue and organismal size requires the continual reprogramming of metabolic pathways to integrate biosynthetic and degradative signals. During cell growth and/or proliferation, one such mechanism that promotes the accumulation of cellular material is a switch from oxidative to glycolytic metabolism, whereby glycolytic intermediates are diverted towards anabolic pathways. How this switch is regulated in different tissues is not clear. Herein we identify a novel role for the tripartite motif (TRIM) family member, TRIM32, in the maintenance of glycolytic flux. Using a proteomics approach, we uncovered the glycolytic enzymes Aldolase (Ald) and Phosphoglycerate mutase 78 (Pglym) as TRIM32 interacting proteins. Loss of Drosophila TRIM32, encoded by the thin (tn) gene, showed a reduction in glycolytic activity and amino acid abundance. This altered metabolic profile caused a striking reduction in the overall size of two inherently glycolytic larval tissues – somatic muscles and the developing brain. Consistent with a role for metabolic intermediates in glycolysis‐driven biomass production, nutritional supplementation of amino acids in tn mutants restored muscle mass. Many tumors favor glycolytic metabolism to maximize substrate production for uncontrolled cell growth and proliferation. Remarkably, wing disc‐associated tumor growth is abolished upon loss of TRIM32. Our results reveal a novel connection between TRIM32 and the maintenance of glycolytic enzyme levels and upregulated pathway activity for the sustained growth of normal and cancerous tissue growth.Support or Funding InformationThis work was supported by a grant through the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) to E.R.G (R01AR060788). J.M.T. is supported by a MIRA award from NIGMS (R35GM119557).
Bringing Anatomy Back
Authors: Helgren Maureen , Giblin Jillian , Nankin Matthew , Grgurich Tania 
Published: April 2020

The paradigm shift from the longstanding “2+2” medical school model to a contemporary curriculum aimed at competencies in clinical reasoning and decision making has challenged medical educators to consider pedagogical and content reform. The integrated curriculum model is an attempt at reform. To date, the outcome of the paradigm shift is uncertain. From a curriculum development view, there is no shortage of desired curricular content; there are time restraints. With the advent of open sources and virtual technologies specific to anatomy, costs of operating human anatomy laboratories, and curricular demands for truncated time dedicated to anatomical foundation, our institution has implemented a variety of curricular and co‐curricular initiatives for multiple learners to engage with anatomy in active learning settings. Most of the initiatives take advantage of collaborative learning between learners from different disciplines and all emphasize the value of understanding cadaveric anatomy from a functional approach to focus on diagnostic reasoning.Examples of three successful curricular initiatives include: 1) Diagnostic imaging of our Silent Mentors. Diagnostic Imaging (DI) students who have completed a Human Anatomy course, prepare CT scans of Silent Mentors (cadaver donors) for students taking Human Anatomy courses in Occupational Therapy (OT), Pathology Assistant (PathA), Physical Therapy (PT), Physician Assistant (PA), and for 1st year medical students in system blocks of the undergraduate medical education (UME) curriculum. The scans are used for horizontal and vertical integration of sectional anatomy. 2) PathA students who have completed a Human Anatomy course return to the lab to partner with 1st year medical students on a 6‐month pathology project. 3) Students taking 4th year electives in medical education or pathology have the option to return to the human anatomy laboratory.Examples of three successful co‐curricular initiatives include: 1) A series of interprofessional case‐based continuing medical education courses with a dissection component. Practicing clinicians and students from medicine, nursing, OT, PA, and PT review a patient case and share discipline‐specific perspectives. 2) IPE60. A peer learning strategy modeled after collegiate athlete “power hours”. Theme‐based series of one‐hour study groups of medical and PT students in the human anatomy lab. 3) Pipeline programs. Anatomy modules delivered by faculty and healthcare profession students interprofessionally or discipline‐specific to local high school and college students.Assessment of curricular initiatives support their utility of meeting learning objectives with a high degree of student satisfaction.
The Use of a Portable Virtual Reality System as a Supplementary Educational Tool in a First‐Year Human Anatomy Course
Authors: Albabish William 
Published: April 2020

Significant advancements in consumer‐grade virtual reality (VR) systems are providing educators with new tools to interact with students. Prior to 2016, VR systems were cost‐prohibitive, limited in software selection, and complex to use. The Oculus Rift, first introduced in 2013, revolutionized the VR field by bringing forth the first affordable six‐degrees of freedom (6‐DoF) commercialized VR system. A functional 6‐DoF VR system consists of the wearable headset, two hand controllers that allow users to interact with objects in the VR space naturally, external sensors to track movements, and requires a costly VR‐ready computer system to render the graphics.In the fall of 2017, a strike affecting all colleges in Ontario limited the ability to deliver onsite lectures for five weeks. During the strike, innovative ways of lecture delivery, such as VR lectures, were successfully implemented to meet the course objectives of a first‐year dissection‐based human anatomy course. However, a notable limitation of VR technology at the time was its need to be tethered to a computer, which confined its use to a small, pre‐defined work area. The limited accessibility of the technology led to a higher amount of time and effort required to create VR‐based educational content. Recent advancements in VR technology led to a new, all‐in‐one 6‐DoF VR system. The Oculus Quest, released in May 2019, incorporates an on‐board computer, built‐in camera sensors, and an on‐board battery. The combination of these features resulted in a powerful, affordable, and portable system that can be used anywhere, anytime, with ease.The Oculus Quest was incorporated into the Fall 2019 first‐year human anatomy course. Videos re‐enforcing course lectures and major anatomical concepts, such as abdominal blood supply, the brachial plexus, and blood supply to the upper limb, were created and distributed digitally to students. Additionally, new software features allow the instructor to export the 3D models created in the VR space as a 3D file, which allows students to view the 3D models on their own VR headsets or computer devices, and 3D print the structures created in the VR space. Preliminary data suggests that students had a better understanding of anatomical concepts, relationships, and depth when using the supplementary VR resources in conjunction with traditional resources. Moreover, VR video content attained a higher viewer retention rate and viewer satisfaction when compared to 2D videos.
Acute and Chronic Laryngeal Deficits Following Cervical Spinal Cord Injury
Authors: Pitts Teresa , Frazure Michael , Strecker Courtney , Greene Clint , Howland Dena , Iceman Kimberly 
Published: April 2020

Laryngeal dysfunction is a common symptom following cervical spinal cord injury (cSCI) and currently there are no effective pharmacological interventions or behavioral treatments. The larynx is the primary valve controlling entrance into the trachea and lungs via opening and closing of the vocal folds. Although not well studied following cSCI, it is understood that laryngeal deficits during swallow often produce aspiration of food/liquid, significantly increasing risk of pneumonia. Our previous terminal electrophysiology experiments using freely breathing sodium pentobarbital anesthetized cats with acute hemisections at the 3rd cervical spinal level (C3) showed immediate alterations in laryngeal activity. These alterations persisted for 4 hours (testing duration). The serotonin (5‐HT1A) receptor agonist 8‐OH‐DPAT was delivered via the vertebral artery at low (3 □g/kg) and/or high (30 □g/kg) doses. This agonist altered behavior‐specific muscle recruitment patterns. In particular, the high dose restored laryngeal activity across breathing and swallow, which could be reversed with the competitive antagonist WAY100635 (0.5 mg/kg). In the current study, we tested if alterations seen acutely in laryngeal activity persist chronically, and if the 8‐OH‐DPAT clinical‐correlate buspirone (Buspar) would produce similar therapeutic results. Adult female cats underwent C3 hemisections and recovered for 4 or 8 weeks. Laryngeal endoscopy was performed weekly under isofluorane (3%) during normocapnia and hypercapnia (5% CO2). Asynchronous vocal fold movement was seen in all animals, defined by unilateral movement delays, and the extent of opening and closing of the glottis became variable. In a subset of animals, oral buspirone (5 mg) was given with food. One hour later, repeat endoscopic evaluation revealed full resolution of laryngeal function with synchronized movements during normocapnia and hypercapnia. Most importantly, with termination of buspirone dosing, laryngeal dysfunction returned. This work shows that the anesthetized preparation effectively identifies upper airway dysfunction early and chronically after C3 SCI, and that it can be used as a pre‐clinical model for novel therapeutic interventions. Results also indicate that buspirone is a high‐priority target for additional investigation as a therapeutic intervention after cSCI.Support or Funding InformationThis work was supported by NIH grants HL 111215, HL 103415 and OT20D001983, the Craig H. Neilsen Foundation Pilot Research Grant 546714, Veterans Affairs Rehabilitation, Research and Development RCSB9249S (DRH), Kentucky Spinal Cord and Head Injury Research Trust, Rebecca F. Hammond Endowment (DRH), and the Commonwealth of Kentucky Challenge for Excellence.
Interactions of Ionophoric Polyphenols with Human Serum Albumin
Authors: Pichardo Rosemary J. , Zahran Mai , Martinez Alberto , Guevara Johnny , Asim Junaid , Gomez Miguel 
Published: April 2020

The distribution of a drug is determined, at least in part, by its interactions with serum proteins. Human serum albumin (HSA), the most abundant blood plasma protein, serves as the most common mode of transport for drugs to their target. In our previous work, we have synthesized and studied important characteristics of a series of ionophoric polyphenols as potential anti‐Alzheimer’s disease agents. As part of our ongoing investigations on the biological activity of these compounds, we are now exploring possible transport mechanisms. In our current work, the interactions of the ionophoric polyphenols with HSA were studied via in vitro and in silico methods. In line with in vitro studies, the interaction of the ionophoric polyphenols with HSA were studied by fluorescence and circular dichroism (CD) spectroscopies to obtain binding affinities, bimolecular quenching constants, theoretical number of binding sites and HSA secondary structural changes. Moreover, in line with in silico studies, a preferred mode of binding was predicted by molecular docking and the predicted systems were subject to a 5 ns MD simulation; consequently, RMSD studies were performed to attest for stability of the systems, MM/GBSA binding free energies were calculated to correlate with experimental binding affinities and per‐residue energy decomposition analyses were carried out to analyze residues critical to binding. Using resveratrol, a polyphenolic compound which has demonstrated strong neuroprotective and anti‐Alzheimer disease properties, as a control, in vitro results showed the formation of adducts (static quenching), one theoretical binding site, binding constants (Ka) ranging 104 to 106 M−1 and only slight secondary structure modifications between HSA and the ionophoric polyphenols. Further, in silico results revealed the ionophoric polyphenols and resveratrol favorably binding subdomain IIA of HSA, forming critical interactions to Trp 214 and Asp451. RMSD studies showed the HSA‐IIA system plateauing after a short time indicating stable binding events; in addition, a Pearson correlation coefficient of R=0.80 between the experimental binding affinity and computational binding free energies showed a strong correlation between in vitro and in silico results. Overall, our results seem to indicate that there is a ionophoric polyphenol‐serum albumin interaction, which could have a potential impact on the use of ionophoric polyphenols as therapeutics for Alzheimer’s disease.Support or Funding InformationThis research used resources of the National Energy Research Scientific Computing Center (NERSC), a U.S. Department of Energy Office of Science User Facility operated under Contract No. DE‐AC02‐05CH11231.