bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neural control of respiratory musculature in the mdx mouse model of Duchenne muscular dystrophy
Authors: Burns David P. , O’Driscoll Eabha , Lucking Eric F. , O’Connor Karen M. , Clarke Gerard , Walter Glenn A. , Edge Deirdre , Fuller David D. , O’Halloran Ken D. 
Published: April 2020

Duchenne muscular dystrophy (DMD) is an inherited fatal neuromuscular disease. Despite deficits in respiratory muscle performance being well recognised, there is a paucity of knowledge in respect of the neural control of breathing in dystrophinopathies. We sought to examine neural control of the diaphragm and upper airway musculature in dystrophin deficient mdx mice.Wild‐type (WT) and mdx mice were studied to examine diaphragm and genioglossus (GG) EMG activity using fine concentric needle electrodes in urethane (1.7mg/kg) anaesthetized spontaneously breathing mice. Basal EMG and responsiveness to chemostimulation were determined. In separate studies, phrenic and hypoglossal (XII) motor output were assessed in anaesthetized, vagotomized, paralyzed and mechanically ventilated WT and mdx mice.Diaphragm and upper airway (sternohyoid) muscle form and function were assessed. Monoamine concentrations, density of glial cells, and pro‐inflammatory gene expression were determined in the spinal cord (C3–C5) and brainstem. Diaphragm and GG EMG activities were depressed in mdx compared to WT during baseline and hypoxic‐hypercapnic challenge. Phrenic and XII nerve recordings revealed enhanced neural drive to diaphragm and GG in mdx mice during hypoxic‐hypercapnic challenge. Excitatory monoamine concentrations were elevated in mdx spinal cord (noradrenaline and serotonin) and brainstem (serotonin). Putative phrenic and hypoglossal motor nuclei in mdx mice had unaltered expression of microglia and astrocytes compared to WT. NFkB and GFAP mRNA expression were increased in mdx brainstem compared to WT, while GFAP mRNA was reduced in mdx spinal cord.Our study revealed a potentiated neural drive in phrenic and XII motor pathways in mdx mice during chemoactivation, suggesting plasticity in motor pathways facilitating breathing. Depressed EMG activity despite potentiated neural outputs, suggests compromised neuromuscular transmission, which we have also observed in respiratory EMG recordings during protracted airway obstruction (Burns et al., 2019, J Physiol). Studies of the fundamental control of breathing in models of DMD should provide a comprehensive vista of the disease, with the potential to identify novel therapeutic targets.
Is Integration Within Anatomical Sciences Important? Assessing Medical Student Learning of Histology and Embryology via Interdisciplinary Causal Mechanisms
Authors: Cholack George , Lisk Kristina , Venuti Judith M. , Attardi Stefanie M. 
Published: April 2020

Curriculum integration is increasingly recognized as a critical component of undergraduate medical education. One approach to curricular integration is through the use of causal mechanisms, which are links that guide students in making purposeful connections between two distinct disciplines. Woods et al. have shown that integrating basic science with clinical content in various contexts using causal mechanisms results in improved diagnostic performance in learners. It remains unknown whether utilizing a similar integrated approach during instruction of different basic sciences, such as embryology and histology, results in better understanding and application of those disciplines. The aim of this study was to determine the effects of integrating histology and embryology using causal mechanisms on medical students’ immediate and delayed (1‐week) recall and application. Second‐year medical students at Oakland University William Beaumont School of Medicine (n=52) participated in this study and were randomized to either an experimental (n=27) or control group (n=25). The experimental group watched a video about pituitary histology and embryology that explicitly integrated these disciplines through causal mechanisms. The control group watched a video covering the same content, but lacked interdisciplinary causal mechanisms. Counterbalanced immediate and delayed post‐tests assessed recall and application of the content (Blooming Anatomy Tool level 1 and 3 questions, respectively). No significant differences (2‐way ANOVA with Bonferroni correction) were observed between the groups on immediate or delayed tests in terms of overall score (p=0.48), histology subscore (p=0.42), embryology subscore (p=0.78), recall subscore (p=0.64), and application subscore (p=0.61). The lack of significant differences between the two learning groups may be due to the close proximity (temporal integration) of embryology and histology instruction afforded to all participants, despite the control group not receiving instruction with causal mechanisms. It is possible that the explicit integration of causal mechanisms does not provide added value when simultaneously learning embryology and histology. Further research is warranted to identify the effect of temporal integration on medical students’ learning of distinct disciplines within the anatomical sciences.Support or Funding InformationOUWB Fellowship in Medical Education
Roles of C‐C motif chemokine ligand (CCL) 2 in aggressive phenotypic change of non‐neoplastic breast cells by tumor‐associated macrophages
Authors: Moon Aree , Lee Seungeun , Lee Eunhye , Ko EunYi , Ham Mina , Lee Hye Min , Kim Eun-Sook , Koh Minsoo , Lim Hyun Kyung , Jung Joohee , Park So Yeon 
Published: April 2020

Tumor‐associated macrophages (TAMs) are major components of tumor microenvironment that promote invasion and metastasis of cancer cells. In this study, we investigated the effect of TAMs on phenotypic conversion of non‐neoplastic MCF10A human breast epithelial cells using an indirect co‐culture system. Co‐culture with TAMs induced epithelial‐to‐mesenchymal transition, invasive phenotype, and MMP‐9 upregulation in MCF10A cells. Comparative proteomic analysis revealed that endoplasmic reticulum oxidoreductase (ERO)1‐α was increased in MCF10A cells co‐cultured with TAMs compared to that in mono‐cultured cells. ERO1‐α was crucial for TAMs‐induced invasive phenotype and MMP‐9 upregulation involving transcription factors c‐fos and c‐Jun. Cytokine array analysis showed that levels of interleukin (IL)‐6, C‐X‐C motif ligand (CXCL)1, C‐C motif ligand (CCL)2, growth‐regulated oncogene (GRO), IL‐8, and granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) were increased in conditioned media of co‐cultured cells. Among these cytokines increased in conditioned media of co‐cultured cells, CCL2 was secreted from TAMs, leading to induction of ERO1‐α, MMP‐9 upregulation, and invasiveness in MCF10A cells. Our findings elucidated a molecular mechanism underlying the aggressive phenotypic change of non‐neoplastic breast cells by co‐culture with TAMs, providing useful information for prevention or treatment of recurrent breast cancer.Support or Funding InformationThe present study was supported by the National Research Foundation of Korea (No. 2013R1A2A2A04013379, No.2015M3A9B6074045, and No. 2016R1A6A1A03007648).
An Organoid Approach to Target Her2/PD‐L1 Positive Gastric Cancers
Authors: Chakrabarti Jayati , Koh Vivien , Chan Shing Leng , Wang Jiang , Ahmed Syed , Ito Yoshiaki , So Jimmy Bok Yan , Yong Wei Peng , Zavros Yana 
Published: April 2020

BackgroundTumors can evade immune surveillance by expressing ligands such as programmed cell death 1 (PDL‐1) which interacts with PD‐1 or CTLA‐4, respectively, on CD8+ cytotoxic T lymphocytes (CTLs). These interactions inhibit CTL proliferation, survival, and effector function subsequently leading to immune evasion and cancer persistence. Despite that 40% of gastric cancers express PD‐L1, the response rate to immunotherapy is only 30%. Human epidermal growth factor receptor2 (HER2) is one of the critical regulators of several metastatic cancers, including gastric. Half of PD‐L1‐positive gastric cancers co‐express HER2, but whether HER2‐induced PD‐L1 sustains tumor cell immune evasion has not been determined.HypothesisThe current research tests the hypothesis that targeted anti‐HER2 and immune checkpoint inhibition may provide synergistic effects in gastric cancer treatment.MethodsPatient‐derived gastric cancer organoids, generated from resected tumor tissues or biopsies were co‐cultured with cytotoxic T lymphocytes (CTLs) and tumor antigenloaded dendritic cells and myeloid‐derived suppressor cells (MDSCs).ResultsWhile PD‐L1‐positive tumor organoids co‐expressed HER2, pharmacological inhibition and gene knockdown of HER2 resulted in decreased PD‐L1 expression in tumor organoids. Within the organoid/immune cell co‐culture, decreased PD‐L1 expression in patient‐derived gastric cancer organoids correlated with a significant increase in CTL proliferation and survival, leading to cancer cell death. Similar results were within the co‐culture were observed in response to Nivolumab treatment. MDSCs within the co‐culture counteracted the inhibitory effects of Her2 inhibition by Mubritinib and Nivolumab.ConclusionThus, HER2‐induced PD‐L1 may contribute to tumor immune evasion. Organoid/immune cell co‐cultures may be used to effectively screen for targeted therapy in individual gastric cancer patients. Therapeutic anti‐HER2 antibody and, immune checkpoint inhibition, as well as depletion of MDSCs, may provide synergistic effects in gastric cancer treatment.Support or Funding Information• NIH NIDDK R01DK083402‐08 (Zavros)NIH 1U19AI116491‐01, (PIs: Weiss and Wells, Zavros Project Leader 1
Dynamic Phosphorylation and Dephosphorylation of CAP1 (Cyclase‐Associated Protein 1) by Antagonistic Signaling through CDK5 and cAMP are Critical for the Protein Functions in Actin Filament Disassembly and Cell Adhesion
Authors: Xiao Yitong , Zhang Haitao , Ramsey Auburn , Karki Uddhab , Akana Lokesh , Xie Jennifer Y. , Xu Jianfeng , Kelly Thomas , Ono Shoichiro , Zhou Guolei 
Published: April 2020

CAP1 (
Cyclase
Associated
Protein 1) is a conserved actin‐regulating protein that enhances actin filament dynamics and also regulates adhesion in mammalian cells. We previously found that phosphorylation at the Ser307/Ser309 tandem site controls its association with cofilin and actin, and is important for CAP1 to regulate the actin cytoskeleton. Here, we report that transient Ser307/Ser309 phosphorylation is required for both CAP1 functions in actin filament disassembly and cell adhesions. Both the phospho‐mimetic and non‐phosphorylatable CAP1 mutants, which resist transition between phosphorylated and dephosphorylated forms, had defects in rescuing the reduced rate of actin filament disassemblyin the CAP1‐knockdown HeLa cells. The phosphor mutants also had defects in alleviating the elevated FAK (
Focal
Adhesion
Kinase) activity and the enhanced focal adhesions in the knockdown cells. In dissecting further phosphor‐regulatory cell signals for CAP1, we found that CDK5 (
Cyclin‐
Dependent
Kinase 5) phosphorylates both Ser307 and Ser309 residues, whereas cAMP signaling induces dephosphorylation at the tandem site, through its effectors PKA (
Protein
Kinase
A) and Epac (
Exchange
proteins directly
activated by
cAMP). No evidence supports involvement of activated protein phosphatase in executing the dephosphorylation downstream of cAMP, whereas preventing CAP1 access to its kinase CDK5 appears to underlie CAP1 dephosphorylation induced by cAMP. Therefore, this study provides direct cellular evidence that transient phosphorylation is required for both CAP1 functions in actin filament turnover and adhesion, and the novel mechanistic insights substantially extend our knowledge on the cell signals that function in concert to regulate CAP1 by facilitating its transient phosphorylation.Support or Funding InformationThis project was supported by an Institutional Development Award(IDeA) from the National Institute of General Medical Sciencesat the National Institutes of Health [Grant number: P20GM103429].
Evaluation of Osteoblastic Gene Expression in Human Mesenchymal Stem Cells
Authors: MacDonald Amber F. , Bow Austin , Masi Thomas , Dhar Madhu 
Published: April 2020

BackgroundOsteoblastic differentiation is a primary pathway to stimulate bone remodeling within regenerative medicine. Human mesenchymal stem cells (hMSCs) are an attractive model for bone regeneration due to their ability to differentiate into osteoblasts, thus promoting an osteogenic environment. With the development of bone biomaterials, it is important to establish a solid baseline of the genes involved in osteoblast differentiation. Once established, the genes regulating osteoblastic differentiation can be compared to any biomaterial product that may potentially speed up the process of bone regeneration. Therefore, the purpose of this study is to examine gene expression as hMSCs undergo osteoblastic differentiation.MethodshMSCs were extracted from human adipose tissue and cultured in DMEM‐F12 medium, 10% FBS. For experimental conditions, cells were cultured in either a growth medium (GM) or an osteogenic induced medium (OIM), supplemented with β‐glycerophosphate, ascorbic‐2‐phosphate, and dexamethasone. Cells were stained with Alizarin Red S staining to confirm calcium deposits of cells cultured in GM or OIM. hMSCs cultured in OIM were extracted for total RNA at days 3, 7, and 21, followed by cDNA synthesis for RT2 Profiler PCR Human Osteogenesis Array (Qiagen, Hilden, Germany). Gene expressions were analyzed using Qiagen Gene Globe software to determine the relative fold change.ResultshMSCs appear to express alkaline phosphatase (ALPL) and bone gammacarboxyglutamate protein (BGLAP) early throughout OIM culture. In contrast, Sp7 expression increases over time, while Sox9 peaks at Day 7 in comparison to the control at Day 3.ConclusionOur data collectively show a change in gene expressions associated with osteoblastic differentiation such as ALPL, BGLAP, Sp7, and SOX9. Sp7, a master osteoblastic transcription factor, increases over time suggesting hMSCs favoring into osteoblastic transformation. Interestingly, Sox9 is another transcription regulator known to commit hMSCs into osteo‐chondrogenic progenitors, and our data show its expression to increase before diminishing near control levels. However, further studies are needed to confirm the genetic pattern of hMSCS undergoing osteoblastic differentiation as well as the upstream mechanisms controlling their activation. This information will be useful in the application of bone biomaterials as a therapeutic treatment for bone regeneration.
Reimagining dissection lab preparation ‐ the role of digital media in anatomy education
Authors: Ng Nicole , Javadian Farnaz , Tse Shirley , Jiang Andy , Doroudi Majid 
Published: April 2020

IntroductionLearning materials provided to students prior to anatomy dissection labs have classically been in the form of written instructions and images from prosections. However, these materials can be difficult to interpret, especially for beginner students. With the advancement of technology, educational curricula are exploring the integration of digital platforms to supplement more traditional teaching methods.GoalTo investigate the utility of a video‐based guide for approaching dissection.MethodsA video dissection guide was created demonstrating the dissection of the superficial back. The video outlines the anatomy, technique, procedure, and includes review questions. The video was made available online through YouTube, along with a feedback survey. A secondary video dissection guide highlighting the dissection of the deep back was subsequently produced and made available in the same avenue.ResultsAfter two months, the superficial back dissection video has received more than 1,400 views. Feedback was received from 68 respondents; a majority of whom were female (63.2%), aged 20–24 (60.3%), and had a current education level of a postgraduate degree or professional degree (54.4%). The majority of respondents agree or strongly agree that the video presented the anatomy in a clear and organized fashion (95.6%), enhanced their learning of the anatomy (100%), familiarized them with dissection tools and how to use them (89.7%), familiarized them with methods and techniques for dissection (92.6%), was more effective than a dissection guide (97.1%), and was more effective for learning anatomy than a textbook (88.2%). A poll of 54 respondents found that most agree or strongly agree that the video is a valuable resource for review/test preparation (88.9%).Preliminary survey results for the deep back dissection video indicates that, of 15 respondents, the majority agree or strongly agree that the video presented the anatomy in a clear and organized fashion (100%), enhanced their learning of the anatomy (93.3%), was more effective than a dissection guide (93.3%), and is a valuable resource for initial learning of dissection/anatomy (100%) and review/test preparation (86.7%).ConclusionIn summary, digital media in the form of video‐based dissection guides may be a useful tool to incorporate into educational curricula for teaching gross anatomy of both superficial and deep structures. Future goals include the incorporation of clinically relevant information to dissected anatomical structures.
Increased activation of HDAC1, 2, and 6, and Sp1 underlies therapeutic resistance and tumor growth in glioblastoma
Authors: Chuang Jian-Ying , Yang Wen-Bin , Hsu Che-Chia 
Published: April 2020

BackgroundGlioblastoma is associated with poor prognosis and high mortality. Although the use of first‐line temozolomide can reduce tumor growth, therapy‐induced stress drives stem cells out of quiescence, leading to chemo‐resistance and glioblastoma recurrence. The Sp1 transcription factor is known to protect glioblastoma cells against temozolomide; however, how tumor cells hijack this factor to gain resistance to therapy is not known.MethodsSp1 acetylation in temozolomide‐resistant cells and stem‐like tumorspheres was analyzed by immunoprecipitation and immunoblotting experiments. Effects of the HDAC/Sp1 axis on malignant growth were examined using cell proliferation–related assays and in vivo experiments. Furthermore, integrative analysis of gene expression with ChIP‐seq as well as the recurrent glioblastoma omics data were also used to further determine the target genes of the HDAC/Sp1 axis.ResultsWe identified Sp1 as a novel substrate of HDAC6, and observed that the HDAC1, 2, 6/Sp1 pathway promotes self‐renewal of malignancy by upregulating BMI1 and hTERT, as well as by regulating G2/M progression and DNA repair via alteration of the transcription of various genes. Importantly, HDAC1, 2, 6/Sp1 activation is associated with poor clinical outcome in both glioblastoma and low‐grade gliomas. However, treatment with azaindolylsulfonamide, a potent HDAC6 inhibitor with partial efficacy against HDAC1 and HDAC2, induced G2/M arrest and senescence in both temozolomide‐resistant cells and stem‐like tumorspheres.ConclusionsOur study uncovers a previously unknown regulatory mechanism in which the HDAC6‐Sp1 axis induces cell division and maintains the stem cell population to fuel tumor growth and therapeutic resistance.Support or Funding InformationThis work was financially supported by research grants from the Ministry of Science and Technology in Taiwan
Pilates Exercise in Hypoxia Increases Vascular Endothelial Function in Professional Pilates Players
Authors: Jung Kyounghwa , Seo Jongbeom , Jung Won-Sang , Park Hun-Young , Lim Kiwon 
Published: April 2020

PurposeThe purpose of this study was to investigate the pilates exercise in hypoxia increases vascular endothelial function in professional Pilates players.MethodsTen healthy Pilates expert female (age: 26.9 ± 2.7, weight: 50.8 ± 5.2, BMI: 19.3 ± 1.4) completed two different trials on different days, consisting of exercise in normoxia (N trial) and exercise in 3,000 m simulated altitude (526 mmHg) hypobaric hypoxia (H trial). After 30 minutes of exposure in each environment, they performed tubing Pilates exercise for 50 minutes under normoxia and hypoxia. Pilates exercise using tubing were composed of 25 types, each performed for 2 minutes. Pilates motion is configured as follows : roll up & down, biceps, arm circles, teaser, rolling, spine twist, mermaid, cobra, swimming, double kicks, swan, cat, thigh stretching, hug a tree, squat, row, saw, hip pull, hundred, lats pull, leg arc, scissor, helicopter, right side leg pull, left side leg pull. Metabolic parameters (minute ventilation; VE, oxygen uptake; VO2, carbon dioxide excretion; VCO2, respiratory exchange ratio; RER, carbohydrate oxidation; CO, fatty acid oxidation; FAO, energy expenditure; EE) and cardiac function (heart rate; HR, stroke volume; SV, cardiac output; CO, end‐diastolic volume; EDV, end‐systolic volume; ESV, ejection fraction; EF) were measured every minutes during Pilates exercise. The sum of the values measured for 50 minutes was used as the result (exceptionally, RER uses average values). Blood pressure (BP), branchial ankle pulse wave velocity (baPWV) and flow mediated dilation (FMD) in branchial artery were measured as vascular function before and after exercise.ResultsIn metabolic parameters, H trial showed significantly higher VE, VCO2, RER and CO than N trial. However, FAO was lower in H trial than in N trial. VO2 and EE. VO2 and EE presented no significant difference between the two trials. In cardiac function, HR showed a significant increase in H trial compared with N trial, however, the remaining cardiac function parameters did not show significant difference. In vascular function, BP (systolic blood pressure, diastolic blood pressure, pulse pressure and mean arterial pressure) and baPWV did not show significant interaction effects between exercise and environment. However, FMD presented a significant interaction effects between exercise and environment. As a result of the post‐hoc, both trial showed a significant increase in FMD via exercise and the increase rate in FMD was significantly higher in H trial (48.5 ± 22.9) than in N trial (34.9 ± 18.3). Also, H trial showed a significantly higher FMD before exercise than N trial.ConclusionOur results suggest that Pilates exercise under moderate hypoxia shows greater metabolic and cardiac response by relatively higher exercise intensity, and it also elicit synergy effect on vascular endothelial function than under normoxia in professional Pilates players.