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    SubjectsResearch Article (132)Biology (75)Medicine (58)Neuroscience (42)Molecular Cell Biology (39)View MoreJournalArsenal: Augusta University's Undergraduate Research Journal (5)Journal of the American Board of Family Medicine : JABFM (4)American family physician (2)International Journal of Clinical Aromatherapy (1)Issues in the Undergraduate Methematics Preparation of School Teachers: The Journal (1)View MoreAuthorsDepartment of Neurology (74)College of Graduate Studies (68)Mei, Lin (43)Department of Cellular Biology and Anatomy (30)Department of Oral Biology (17)View MoreTypes
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    Epigenetic mechanisms and genome stability

    Putiri, Emily L.; Robertson, Keith D. (2010-12-29)
    Keywords: DNA methylation
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    Modulation of Dnmt3b function in vitro by interactions with Dnmt3L, Dnmt3a and Dnmt3b splice variants

    Van Emburgh, Beth O.; Robertson, Keith D. (2011-07-4)
    DNA methylation, an essential regulator of transcription and chromatin structure, is established and maintained by the coordinated action of three DNA methyltransferases: DNMT1, DNMT3A and DNMT3B, and the inactive accessory factor DNMT3L. Disruptions in DNMT3B function are linked to carcinogenesis and genetic disease. DNMT3B is also highly alternatively spliced in a tissue- and disease-specific manner. The impact of intra-DNMT3 interactions and alternative splicing on the function of DNMT3 family members remains unclear. In the present work, we focused on DNMT3B. Using a panel of in vitro assays, we examined the consequences of DNMT3B splicing and mutations on its ability to bind DNA, interact with itself and other DNMT3's, and methylate DNA. Our results show that, while the C-terminal catalytic domain is critical for most DNMT3B functions, parts of the N-terminal region, including the PWWP domain, are also important. Alternative splicing and domain deletions also influence DNMT3Bâ s cellular localization. Furthermore, our data reveal the existence of extensive DNMT3B self-interactions that differentially impact on its activity. Finally, we show that catalytically inactive isoforms of DNMT3B are capable of modulating the activity of DNMT3Aâ DNMT3L complexes. Our studies therefore suggest that seemingly â inactiveâ DNMT3B isoforms may influence genomic methylation patterns in vivo.
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    NOMA: A Preventable "Scourge" of African Children

    Ogbureke, Kalu U.E.; Ogbureke, Ezinne I (2010-10-21)
    Noma is a serious orofacial gangrene originating intraorally in the gingival-oral mucosa complex before spreading extraorally to produce a visibly destructive ulcer. Although cases of noma are now rarely reported in the developed countries, it is still prevalent among children in third world countries, notably in sub-Sahara Africa, where poverty, ignorance, malnutrition, and preventable childhood infections are still common. This review summarizes historical, epidemiological, management, and research updates on noma with suggestions for its prevention and ultimate global eradication. The global annual incidence remains high at about 140,000 cases, with a mortality rate exceeding 90% for untreated diseases. Where the patients survive, noma defects result in unsightly facial disfigurement, intense scarring, trismus, oral incompetence, and social alienation. Although the etiology has long been held to be infectious, a definitive causal role between microorganisms cited, and noma has been difficult to establish. The management of noma with active disease requires antibiotics followed by reconstructive surgery. Current research efforts are focused towards a comprehensive understanding of the epidemiology, and further elucidation of the microbiology and pathogenesis of noma.
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    Prediction of diabetic retinopathy: role of oxidative stress and relevance of apoptotic biomarkers

    Al-Shabrawey, Mohamed; Smith, Sylvia B (2010-03-23)
    Keywords: Diabetic retinopathy
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    NF-Y Recruits Both Transcription Activator and Repressor to Modulate Tissue- and Developmental Stage-Specific Expression of Human γ-Globin Gene

    Zhu, Xingguo; Wang, Yongchao; Pi, Wenhu; Liu, Hui; Wickrema, Amittha; Tuan, Dorothy (2012-10-10)
    The human embryonic, fetal and adult β-like globin genes provide a paradigm for tissue- and developmental stage-specific gene regulation. The fetal γ-globin gene is expressed in fetal erythroid cells but is repressed in adult erythroid cells. The molecular mechanism underlying this transcriptional switch during erythroid development is not completely understood. Here, we used a combination of in vitro and in vivo assays to dissect the molecular assemblies of the active and the repressed proximal γ-globin promoter complexes in K562 human erythroleukemia cell line and primary human fetal and adult erythroid cells. We found that the proximal γ-globin promoter complex is assembled by a developmentally regulated, general transcription activator NF-Y bound strongly at the tandem CCAAT motifs near the TATA box. NF-Y recruits to neighboring DNA motifs the developmentally regulated, erythroid transcription activator GATA-2 and general repressor BCL11A, which in turn recruit erythroid repressor GATA-1 and general repressor COUP-TFII to form respectively the NF-Y/GATA-2 transcription activator hub and the BCL11A/COUP-TFII/GATA-1 transcription repressor hub. Both the activator and the repressor hubs are present in both the active and the repressed γ-globin promoter complexes in fetal and adult erythroid cells. Through changes in their levels and respective interactions with the co-activators and co-repressors during erythroid development, the activator and the repressor hubs modulate erythroid- and developmental stage-specific transcription of γ-globin gene.
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    Multicolor Time-lapse Imaging of Transgenic Zebrafish: Visualizing Retinal Stem Cells Activated by Targeted Neuronal Cell Ablation

    Ariga, Junko; Walker, Steven L.; Mumm, Jeff S. (2010-09-20)
    High-resolution time-lapse imaging of living zebrafish larvae can be utilized to visualize how biological processes unfold (for review see 1). Compound transgenic fish which express different fluorescent reporters in neighboring cell types provide a means of following cellular interactions 2 and/or tissue-level responses to experimental manipulations over time. In this video, we demonstrate methods that can be used for imaging multiple transgenically labeled cell types serially in individual fish over time courses that can span from minutes to several days. The techniques described are applicable to any study seeking to correlate the "behavior" of neighboring cells types over time, including: 1) serial 'catch and release' methods for imaging a large number of fish over successive days, 2) simplified approaches for separating fluorophores with overlapping excitation/emission profiles (e.g., GFP and YFP), 3) use of hypopigmented mutant lines to extend the time window available for high-resolution imaging into late larval stages of development, 4) use of membrane targeted fluorescent reporters to reveal fine morphological detail of individual cells as well as cellular details in larger populations of cells, and 5) a previously described method for chemically-induced ablation of transgenically targeted cell types; i.e., nitroreductase (NTR) mediated conversion of prodrug substrates, such as metronidazole (MTZ), to cytotoxic derivatives 3,5.
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    Techniques Used to Establish the First Person Narrator and Perspective in Double Indemnity and Murder, My Sweet

    Walton, Breana (Augusta University Libraries, 2017-05-11)
    Directed by Billy Wilder and Edward Dmytryk respectively, the films noir Double Indemnity (1944) and Murder, My Sweet (1944) each have a storyline that unfolds from a first person perspective as told by a narrator. The techniques used in the films establish this first person perspective through which the films are understood. Both films include voice over as a technique, which determines who the narrator is and the amount of information withheld or disclosed to the audience. Establishing the visual perspective of the narrator is portrayed through differently for each film. While, Double Indemnity utilizes camera angle, Murder, My Sweet uses camera filters and special effects. Lastly, to achieve the first person narration, the character narrating in each film must be present in every scene or give explanation of events that occur in his absence. The various techniques used in each film function cohesively to establish the narrator and achieve his perspective through which the plot is understood by the audience.
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    Studying Gene Expression in Whole Embryos by in situ Hybridization: A Peer-to-Peer Laboratory Guide

    Kalra, Aarushi; Xia, Di (Augusta University Libraries, 2017-05-11)
    The extracellular matrix (ECM) plays an important role in cell to cell signaling pathways. Our goal is to provide a full laboratory guide for students to study gene expression in zebrafish embryos by in situ hybridization. Prior to our study, the laboratory had observed disorganized and shortened cilia in cells that are important for cell signaling in the pronephric duct and neural tube floor plate of the zebrafish embryo. Ciliogenesis depends on a master transcriptional regulator, foxj1a, whose mRNA expression can be monitored through in situ hybridization and microscopic imaging. Knockdown morpholino-injected, control mismatched morpholino-injected, and uninjected embryos were fixed to determine if foxj1a transcription is qualitatively affected by ECM gene knockdown. Our results showed that the knockdown embryos portrayed an inconsistent foxj1a signal strength along the length of the pronephric duct, when compared to analysis of control mismatched and wild-type uninjected embryos. We created this manuscript for other students to observe how ECM gene knockdown can affect foxj1a mRNA expression, but also to give them a guide to the tools they would need to explore their own genes of interest, in zebrafish or in many other organisms and tissues.
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    Binge-Pattern Alcohol Exposure during Puberty Induces Long-Term Changes in HPA Axis Reactivity

    Przybycien-Szymanska, Magdalena M.; Mott, Natasha N.; Paul, Caitlin R.; Gillespie, Roberta A.; Pak, Toni R. (2011-04-13)
    Adolescence is a dynamic and important period of brain development however, little is known about the long-term neurobiological consequences of alcohol consumption during puberty. Our previous studies showed that binge-pattern ethanol (EtOH) treatment during pubertal development negatively dysregulated the responsiveness of the hypothalamo-pituitary-adrenal (HPA) axis, as manifested by alterations in corticotrophin-releasing hormone (CRH), arginine vasopressin (AVP), and corticosterone (CORT) during this time period. Thus, the primary goal of this study was to determine whether these observed changes in important central regulators of the stress response were permanent or transient. In this study, juvenile male Wistar rats were treated with a binge-pattern EtOH treatment paradigm or saline alone for 8 days. The animals were left undisturbed until adulthood when they received a second round of treatments consisting of saline alone, a single dose of EtOH, or a second binge-pattern treatment paradigm. The results showed that pubertal binge-pattern EtOH exposure induced striking long-lasting alterations of many HPA axis parameters. Overall, our data provide strong evidence that binge-pattern EtOH exposure during pubertal maturation has long-term detrimental effects for the healthy development of the HPA axis.
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    Lactate Produced by Glycogenolysis in Astrocytes Regulates Memory Processing

    Newman, Lori A.; Korol, Donna L.; Gold, Paul E. (2011-12-13)
    When administered either systemically or centrally, glucose is a potent enhancer of memory processes. Measures of glucose levels in extracellular fluid in the rat hippocampus during memory tests reveal that these levels are dynamic, decreasing in response to memory tasks and loads; exogenous glucose blocks these decreases and enhances memory. The present experiments test the hypothesis that glucose enhancement of memory is mediated by glycogen storage and then metabolism to lactate in astrocytes, which provide lactate to neurons as an energy substrate. Sensitive bioprobes were used to measure brain glucose and lactate levels in 1-sec samples. Extracellular glucose decreased and lactate increased while rats performed a spatial working memory task. Intrahippocampal infusions of lactate enhanced memory in this task. In addition, pharmacological inhibition of astrocytic glycogenolysis impaired memory and this impairment was reversed by administration of lactate or glucose, both of which can provide lactate to neurons in the absence of glycogenolysis. Pharmacological block of the monocarboxylate transporter responsible for lactate uptake into neurons also impaired memory and this impairment was not reversed by either glucose or lactate. These findings support the view that astrocytes regulate memory formation by controlling the provision of lactate to support neuronal functions.
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