bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of the amino acids associated with the novel ribonuclease activity of cusativin via protein engineering and LC‐MS
Authors: Thakur Priti , Atway Jowad , Limbach Patrick A. , Addepalli Balasubrahmanyam 
Published: April 2020

Ribonucleases (RNases) are widely used in detecting the location of chemical modification in RNA, an approach commonly referred to as, “RNA modification mapping”
[1]. The commonly employed RNases include RNase T1 (guanosine‐specific), U2 (purine‐specific) and A (pyrimidine specific). Recently, uridine‐specific (MC1)
[2], cytidine‐specific (cusativin)
[3] and a non‐specific RNase U2
[4] were identified as potential tools for RNA modification mapping. Of these, RNase cusativin exhibits novel cleavage behavior where it does not cleave the bond between two consecutive cytidine nucleotides (CpC)
[3]. This unique feature of cusativin has encouraged us to probe the nucleobase recognition characteristics of the enzyme binding site, which is composed of B1 and B2 sites. We employed amino acid substitutions (through protein engineering), and liquid chromatography coupled with mass spectrometry (LC‐MS) to analyze the contributions of specific amino acids in the enzyme binding site. The mutant proteins generated by specific amino acid substitutions were investigated for alteration of secondary structure, kinetic changes in enzyme activity, and substrate specificity. While single amino acid substitutions revealed minimal effects on the secondary structure, multiple substitutions resulted in significant changes as revealed by circular dichroism studies. However, the mutant proteins, irrespective of the number of substitutions, exhibited remarkable changes in their kinetic behavior with the tested RNA substrate. The LC‐MS analysis of enzymatic digestion products of RNA indicated altered nucleobase specificity depending on the location of amino acid substitutions. While mutation at one location (V99L) behaved more or less like wild type protein, mutation at different position (N97S) but within same B2 binding site pocket revealed generation of digestion products indicative of CpC bond cleavage. However, the N97S mutant also exhibited digestion products that suggest cleavage of RNA at 3′‐end of guanosine. Another mutant protein with amino acid substitutions in both B1 (R69T) and B2 (V99L and T100R) sites also exhibited significant cleavage of CpC bond. But this behavior is also accompanied with more or less equal affinity towards C and G for RNA cleavage. The implications of this preliminary experimental data on the role of specific amino acids in nucleobase recognition of RNA substrate will be discussed.Support or Funding InformationFinancial support of this work was provided by the National Institutes of Health (GM58843), and the University of Cincinnati.
Longitudinal In Vivo Imaging of Atherosclerotic Disease Development in The apoE Deficient Zucker Rat
Authors: Alvidrez Roberto Ivan Mota , Buglak Nicholas , Anderson Tamara , Norenberg Jeffrey , Bahnson Edward 
Published: April 2020

One obstacle hindering the development of therapies to treat diabetic vasculopathies, is access to an affordable and appropriate small animal model that recapitulates human pathology. There are currently no well‐established small animal models to study diabetes and atherosclerosis combined. Longitudinal in vivo imaging of inflammation in atherosclerosis is a promising tool to evaluate progression of cardiovascular complications. However, vascular size of current mouse models of atherosclerosis does not allow for detailed molecular imaging. Radiolabeled DANBIRT has been used as an in vivo molecular imaging tool on environmental exposure, aging, and atherosclerosis. Our group has modified the lipid metabolism in Lean Zucker (LZ: Lepr+/?) rats to promote dyslipidemia and development of atherosclerosis by knocking out the apolipoprotein E (apoE) gene using CRISPR/Cas9. We hypothesize that apoE knockout, LZ rats (apoE−/−, Lepr+/?) will develop atherosclerosis with dietary exposure to high fat, high cholesterol (HFHC) diet. [111In]‐DANBIRT will be able to localize inflammation in vivo and allow longitudinal monitoring of vascular atherosclerotic disease using SPECT/CT imaging.MethodsapoE−/−;LZ adult rats were imaged for 1hr post injection of ~900 μCi of [111In]‐DANBIRT for SPECT/CT imaging using a NanoSPECT/CT®. Rats were placed on either HFHC or standard rodent chow for 10 weeks starting at 12 weeks of age (baseline). apoE+/+;LZ under normal and HFHC diet were used as controls. Regions of Interests (ROI) are identified and determined for muscle (background tissue), carotid arteries, aortic arch, and descending aorta. Activity is decay corrected and compared between and among experimental groups. Serum chemistry and histologic analysis of en face aortas and aortic root were performed. Data is represented as mean ± SEM. Two‐way ANOVA with Tukey’s correction was performed.ResultsAll apoE−/− rats developed severe hypercholesterolemia particularly under a HFHC diet. After 10 weeks of HFHC diet apoE−/−, LZ rats have higher [111In]‐DANBIRT uptake in the descending aorta uniquely compared to baseline (p<0.05) and to normal diet fed rats (ns). Histologic analysis showed apoE−/−, LZ have increased significant atherosclerosis on the aortas and the aortic sinus, which correlates to our imaging results. apoE+/+, LZ rats did not develop atherosclerosis disease as expected.ConclusionTo our knowledge, we have the first polygenic rat model able to develop atherosclerosis translationally under HFHC. [111In]‐DANBIRT longitudinally evidenced atherosclerotic disease in the abdominal aorta in vivo correlated to ex vivo histologic analysis.Support or Funding InformationSupported by the NORC PPF (P30 DK056350) to EMB and the CRTG (Burroughs Wellcome Fund) to RIM.
Establishing Normative Embryonic and Early Postnatal Cranial Bone Volume and Density Values for C57BL/6J Laboratory Mice
Authors: Lesciotto Kate M. , Tomlinson Lauren , Richtsmeier Joan 
Published: April 2020

We provide normative data for cranial bone volume and tissue mineral density (TMD) for embryonic and early postnatal C57BL/6J mice that can be used as control data for researchers using experimental mouse models. C57BL/6J mice were selected based on their common usage in experimental research. Our sample includes 3 males and 3 females at four ages: embryonic day (E) 15.5, E17.5, postnatal day (P) 0, and P7. MicroCT images were acquired by the Center for Quantitative Imaging at the Pennsylvania State University using the General Electric v|tom|x L300 nano/microCT system. Prior to data collection, all postcranial bones were electronically removed from the scan using Avizo 9.4, and then background noise was removed from each scan using a 3D median filter. Isosurfaces were reconstructed to represent cranial bone based on a hydroxyapatite phantom that was scanned with each specimen. To maintain consistency between specimens and across age groups, we used 85 mg/cm3 as the minimum threshold to create isosurfaces and segment bone for this study. Twenty cranial bones were individually segmented for each specimen, and volume and TMD were estimated in Avizo. After correcting for multiple testing, no significant differences were found in bone volume or TMD between males and females for any of the examined cranial bones at any age, and data were pooled for analyses. As expected, volume of ossified bone increased with age from E15.5 to P7. The number of ossification centers present in a single individual is highly variable at E15.5 (ranging from 2 – 10), with all bones present by P7. Most of the measured bones consistently increased in TMD with age, with several exceptions. The palatine, vomer, and zygomatic bones each decreased in TMD from P0 to P7. The TMD of the maxilla also decreased slightly from E17.5 to P0, but then increased from P0 to P7. No significant patterns of development related to originating cell population (mesoderm v. neural crest) or mode of ossification (endochondral v. intramembranous) were apparent. These data can serve as a reference standard for future studies that examine craniofacial growth and disease models using mice bred on a C57BL/6J background.Support or Funding InformationNational Science Foundation Grant BCS‐1731909, NIDCR R01DE027677 and R01 DE022988, and the Pennsylvania State University Energy and Environmental Sustainability Laboratories Green Seed Grant
Advancing Anatomy Education Competencies—A Critical Interpretive Synthesis from Five Key Papers
Authors: Pacey Lindsay , Kulesza Randy , Lomiguen Christine , Terrell Mark 
Published: April 2020

IntroductionSince the early 1900s, time devoted to studying anatomy in medical schools has substantially decreased. Although the number of students enrolled in health professions education programs is increasing, a shortage of formally trained gross anatomists persists. Consequently, anatomy education and learning must be efficient and effective. Programs training the next generation of anatomy educators need to be sure that graduates obtain particular knowledge and skill sets. Competency‐based education (CBE) may provide one such approach. Extensive research on competency‐based education (CBE) exists in medical education that provides developmental milestones and entrustable professional activities (EPAs). A similar process involving CBE and EPAs would benefit anatomy education program development graduates of such programs, students of teachers from such programs, and ultimately patients.AimThe purpose of this research is to identify five key papers in CBE that are most influential for providing a foundation for developing competencies in training the next generation of anatomists as educational specialists.MethodsCritical interpretive synthesis (CIS) will be utilized to identify, summarize, and integrate five key papers into a primer for emerging anatomy educators on competencies highlighted in literature and through professional societies. A comprehensive search of PubMed/MEDLINE, CINAHL, and ERIC databases published in English from 1990–2020 utilizing terms including competency‐based education, faculty development, and anatomy education. Additional searches through relevant journal article’s bibliographies will also be conducted. Papers’ relevance will be given a numerical value based on relevance and usability. A student learning community will be established to generate a cumulative list of the 20 most highyielding papers regarding anatomy education and CBE. These numerical results will go through several iterative processes, inspired by the Delphi methodology, to generate a top five ranked articles that will then be analyzed and integrated into a single CIS paper for publication.ResultsMain concepts from the five key CBE papers will be amalgamated into one primer through CIS to lay the groundwork for developing competencies for future anatomists as education specialists.Discussion and ConclusionResults of this study will aid in helping anatomy educators attain competency and aid in developing and designing curricula to train the next generation of anatomy educators.
Endothelial focal adhesion kinase contributes to intestinal microvascular leakage during ischemia/reperfusion injury
Authors: Eitnier Rebecca A. , Wang Fang , Yang Clement , Aponte Alexandra M. , Ewing Thomas J. , Wu Mack H. 
Published: April 2020

Ischemia/reperfusion (I/R) injury triggers a series of inflammatory responses in the microcirculation characterized by vascular leakage and leukocyte activation leading to organ dysfunction. In this study, we examine whether focal adhesion kinase (FAK) contributes to endothelial barrier dysfunction in response to I/R in splanchnic tissues. Using an in vivo model of intestinal I/R injury, we showed that the increase of microvascular leakage induced by I/R injury was greatly attenuated in endothelial specific, tamoxifen inducible, FAK knockout animals as well as wild type animals given pharmacological inhibition of FAK with PF‐573228. Using murine intestinal endothelial cells, we performed electric cell‐substrate impedance sensing (ECIS) assays with or without FAK inhibitor in the presence of H2O2. Results showed that FAK inhibition significantly attenuated endothelial barrier dysfunction induced by H2O2. Western blot analysis showed that H2O2 promoted FAK phosphorylation in a time course similar to that of endothelial barrier dysfunction. Confocal imaging analysis demonstrated that H2O2 increases the disruption of junctional proteins. The effect was blocked by FAK inhibitor. Taken together, the results suggested that FAK plays an important role in mediating intestinal vascular leakage during I/R injury.Support or Funding InformationNIH HL120954, VA BX000799
Induction of Labour at 41 Weeks Versus Expectant Management and Induction of Labour at 42 Weeks (SWEdish Post-Term Induction Study, SWEPIS): Multicentre, Open Label, Randomised, Superiority Trial
Authors: Wennerholm Ulla-Britt , Saltvedt Sissel , Wessberg Anna , Alkmark Mårten , Bergh Christina , Brismar Wendel Sophia , Fadl Helena , Jonsson Maria , Ladfors Lars , Sengpiel Verena , Wesstrom Jan , Wennergren Göran , Wikstrom Anna-Karin , Elden Helen , Stephansson Olof , Hagberg Henrik 
Published: April 2020

(Abstracted from BMJ 2019;367:l6131)
Adverse perinatal outcomes such as stillbirth and maternal complications have been shown to gradually increase postterm (≥42 weeks [≥294 days]). Despite this, no international agreement exists on how to manage late-term pregnancies (although the World Health Organization does recommend induction of labor at 41 weeks).
American Urogynecologic Society Systematic Review: The Impact of Weight Loss Intervention on Lower Urinary Tract Symptoms and Urinary Incontinence in Overweight and Obese Women
Authors: Yazdany Tajnoos , Jakus-Waldman Sharon , Jeppson Peter C. , Schimpf Megan O. , Yurteri-Kaplan Ladin A. , Ferzandi Tanaz R. , Weber-LeBrun Emily , Knoepp Leise , Mamik Mamta , Viswanathan Meera , Ward Renée M. 
Published: April 2020

(Abstracted from Female Pelvic Med Reconstr Surg 2020;26:16–29)
Urinary incontinence (UI) affects many women across the world and can have severe impacts on quality of life. Obesity has been reported to play a role in the development of UI.
How Women Are Treated During Facility-Based Childbirth in Four Countries: A Cross-sectional Study With Labour Observations and Community-Based Surveys
Authors: Bohren Meghan A. , Mehrtash Hedieh , Fawole Bukola , Maung Thae Maung , Balde Mamadou Dioulde , Maya Ernest , Thwin Soe Soe , Aderoba Adeniyi K. , Vogel Joshua P. , Irinyenikan Theresa Azonima , Adeyanju A. Olusoji , Mon Nwe Oo , Adu-Bonsaffoh Kwame , Landoulsi Sihem , Guure Chris , Adanu Richard , Diallo Boubacar Alpha , Gülmezoglu A. Metin , Soumah Anne-Marie , Sall Alpha Oumar , Tunçalp Özge 
Published: April 2020

(Abstracted from Lancet 2019;394:1750–1763)
Women around the world experience mistreatment during childbirth. This includes, but is not limited to, physical abuse, verbal abuse, discrimination, and procedures performed without consent.
Adherence of Cell-Free DNA Noninvasive Prenatal Screens to ACMG Recommendations
Authors: Skotko Brian G. , Allyse Megan A. , Bajaj Komal , Best Robert G. , Klugman Susan , Leach Mark , Meredith Stephanie , Michie Marsha , Stoll Katie , Gregg Anthony R. 
Published: April 2020

(Abstracted from Genet Med 2019;21:2285–2292)
Noninvasive prenatal screening (NIPS) with analysis of cell-free DNA (cfDNA) is a procedure that tests for fetal aneuploidies. This testing has been available to women in the United States since 2011, and in 2016, the American College of Medical Genetics and Genomics (ACMG) released specific recommendations for laboratories to follow when administering this testing and reporting results.
Dual Antiplatelet Therapy and Surgical Timing in Geriatric Hip Fracture
Authors: Tarrant Seth M. , Kim Raymond G. , McGregor Kassandra L. , Palazzi Kerrin , Attia John , Balogh Zsolt J. 
Published: April 2020

Objective:
To determine how timing of surgery affects transfusion, major complications, and mortality in patients who sustain a geriatric hip fracture while taking dual antiplatelet therapy (DAPT; typically aspirin and clopidogrel).

Design:
Retrospective cohort study.

Setting:
University-affiliated Level 1 Trauma Center.

Patients:
Patients 65 years of age or older on DAPT with a geriatric hip fracture were investigated at a single institution between 2002 and 2017. Demographic and perioperative data were collected from patient records, institutional databases, and national hip fracture registry.

Intervention:
Fixation or arthroplasty.

Main Outcome Measurement:
Transfusion, major complications, and 30-day mortality.

Results:
Of the 6724 patients sustaining a geriatric hip fracture, 122 patients were taking DAPT on admission. Timing of surgery did not influence transfused units (incidence rate ratio 1.00, 95% confidence interval: 0.87–1.15, P = 0.968) but did affect major complications (time modeled as quadratic term; odds ratios ranging from 0.20 to 7.91, ptime = 0.001, ptime*time<0.001) and 30-day mortality (odds ratio 1.32, 95% confidence interval: 1.03–1.68, P = 0.030).

Conclusion:
Surgical delay does not change the need for transfusion of hip fracture patients on DAPT, but it is associated with increased probabilities of major complications and 30-day mortality.

Level of Evidence:
Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.
Morbidity and Mortality in Critically Ill Children. II. A Qualitative Patient-Level Analysis of Pathophysiologies and Potential Therapeutic Solutions*
Authors: Meert Kathleen L. , Banks Russell , Holubkov Richard , Pollack Murray M. ,   and the Eunice Kennedy Shriver National Institute of Child Health and Human Development Collaborative Pediatric Critical Care Research Network
Published: April 2020

Objectives:
To describe at the individual patient level the pathophysiologic processes contributing to morbidity and mortality in PICUs and therapeutic additions and advances that could potentially prevent or reduce morbidity and mortality.

Design:
Qualitative content analysis of intensivists’ conclusions on pathophysiologic processes and needed therapeutic advances formulated by structured medical record review.

Setting:
Eight children’s hospitals affiliated with the Eunice Kennedy Shriver National Institute of Child Health and Human Development Collaborative Pediatric Critical Care Research Network.

Patients:
A randomly selected cohort of critically ill children with a new functional morbidity or mortality at hospital discharge. New morbidity was assessed using the Functional Status Scale and defined as worsening by two or more points in a single domain from preillness baseline.

Interventions:
None.

Measurements and Main Results:
Of 292 children, 175 (59.9%) had a new morbidity and 117 (40.1%) died. The most common pathophysiology was impaired substrate delivery (n = 158, 54.1%) manifesting as global or regional hypoxia or ischemia due to low cardiac output or cardiac arrest. Other frequent pathophysiologies were inflammation (n = 104, 35.6%) related to sepsis, respiratory failure, acute respiratory distress syndrome, or multiple organ dysfunction; and direct tissue injury (n = 64, 21.9%) including brain and spinal cord trauma. Chronic conditions were often noted (n = 156, 53.4%) as contributing to adverse outcomes. Drug therapies (n = 149, 51.0%) including chemotherapy, inotropes, vasoactive agents, and sedatives were the most frequently proposed needed therapeutic advances. Other frequently proposed therapies included cell regeneration (n = 115, 39.4%) mainly for treatment of neuronal injury, and improved immune and inflammatory modulation (n = 79, 27.1%).

Conclusions:
Low cardiac output and cardiac arrest, inflammation-related organ failures, and CNS trauma were the most common pathophysiologies leading to morbidity and mortality in PICUs. A research agenda focused on better understanding and treatment of these conditions may have high potential to directly impact patient outcomes.
Expiratory Airflow Limitations on Lung Flute Effectiveness in Secretion Clearance: An Observational Cross-sectional Pilot Study
Authors: da Silva Francisco Carlos Nunes , Stolfi Angela , D'Agati Michael 
Published: April 2020

Purpose:
The purpose of this observational study was to assess the validity of the Lung Flute® (LF) manufacturer's criterion for effective use of the device.

Methods:
Peak expiratory flow (PEF) and the ability to activate the oscillatory mechanism in the LF were assessed and compared. The diagnostic accuracy of the manufacturer's PEF threshold value of ≥128.4 L/min was determined, and an optimized threshold was derived.

Results:
The LF manufacturer's PEF threshold value of ≥128.4 L/min had a sensitivity of 100%, a specificity of 55%, and overall diagnostic accuracy of 83%. Nine (45%) of the twenty subjects that were not capable of activating the oscillatory device had a PEF ≥128.4 L/min. The optimal threshold value, with a sensitivity and specificity of 100%, for our sample was 167.5 L/min.

Conclusions:
The manufacturer's PEF threshold value of ≥128.4 L/min demonstrated high sensitivity but low specificity for predicting a patient's ability to use the LF device effectively. The optimized threshold value derived from our study was markedly higher. Clinicians should be aware that the LF manufacturer's specifications for the minimum PEF required for effective use of the device may not be evidence-based or accurate and consider this when prescribing this oscillatory positive expiratory pressure device.
Research on the Radiotoxicology of Plutonium Using Animals: Consideration of the 3Rs—Replace, Reduce, Refine
Authors: Griffiths Nina M. , Van der Meeren Anne , Angulo Jaime F. , Vincent-Naulleau Silvia 
Published: April 2020

Abstract
To characterize the health effects of incorporated plutonium, many experiments have been conducted using different animal models. These range from (1) applied (tissue uptake/retention determination, decorporation therapy efficacy), (2) fundamental (gene expression, cancer induction), and (3) dosimetry models. In recent years, the use of animals for scientific purposes has become a public concern. The application of the 3Rs - Replace (use of alternative methods or animals not considered capable of experiencing pain, suffering, and distress), Reduce (reduction in animal numbers), and Refine (better animal welfare and minimization of suffering, pain and distress) - has increased to address ethical concerns and legislative requirements. The introduction of novel non-animal technologies is also an important factor as complementary options to animal experimentation. In radiotoxicology research, it seems there is a natural tendency to Replace given the possibility of data reuse obtained from contamination cases in man and animal studies. The creation of “registries” and “repositories” for nuclear industry workers (civil and military) is now a rich legacy for radiotoxicological measurements. Similarly, Reduction in animal numbers can be achieved by good experimental planning with prior statistical analyses of animal numbers required to obtain robust data. Multiple measurements in the same animal over time (external body counting, excreta collection) with appropriate detection instruments also allow Reduction. In terms of Refinement, this has become “de rigueur” and a necessity given the societal and legal concerns for animal welfare. For research in radiotoxicology, particularly long-term studies, better housing conditions within the constraints of radiation protection issues for research workers are an important concern. These are all pertinent considerations for the 3Rs remit and future research in radiotoxicology.