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Investigating Roles Of 2 Novel Eklf Targets Involved In Erythropoiesis, Rose M. Gott

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Erythrocytes are primarily comprised of the oxygen carrying protein hemoglobin. Genetic mutations causing defects in the proper synthesis of hemoglobin result in various anemias. It is during the last phases of terminal erythroid differentiation that hemoglobin levels rise, making it a focus for therapeutic research. Fetal hemoglobin is comprised of ⍺-globin and gamma globin, then after a change in gene expression called hemoglobin switching which takes place after birth, adult hemoglobin is comprised of ⍺-globin and beta globin. We investigated hemoglobin switching and erythroid terminal differentiation by focusing on the master erythroid transcription factor Erythroid Krüppel-like Factor (EKLF). Data led …

Development And Analysis Of Next-Generation Polymeric And Bio-Ceramic Based Orthopedic Scaffolds By Advanced Manufacturing Techniques, Sudeep K. Gummadi

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Gliomas express mutant isocitrate dehydrogenases producing excessive amounts of D 2-hydroxyglutarate (D2HG) and releasing some of it into the environment. The immune surveillance is reduced as a result, however, the mechanisms behind lymphocyte suppression by the D2HG stereoisomer remain unknown. I incubated Jurkat T cells with D2HG at concentrations present within and surrounding gliomas, or its obverse L2HG stereoisomer, and quantified 2HG isomers within washed cells by TSPC derivatization with stable isotope-labeled D2HG and L2HG internal standards, HPLC separation, and mass spectrometry. D2HG was found in quiescent cells in double the amount of L2HG. External D2HG or L2HG increased the …

A Multi-Spatial Analysis Of Land Use Effects On Freshwater Mussels In The Upper Cuyahoga River And Tinkers Creek, Tamar Atwell

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Mussels are considered one indicator of good water quality in rivers, but over the past 20 years mussel populations have continued to decline, while water quality improves. According to the Ohio Environmental Protection Agency (OEPA), water quality in the Cuyahoga River is now within safe levels for all parameters. However, what are safe levels for humans may not be safe for mussels. An analysis of freshwater mussel populations in two similarly sized watersheds, the Upper Cuyahoga River and Tinkers Creek was conducted. Correlations of mussel abundance and diversity were assessed by multivariate GIS/remote sensing tools to contrast water flow rates, …

Basal Signaling Through Death Receptor 5 And Caspase 3 Activates P38 Kinase To Regulate Serum Response Factor - Mediated Myod Transcription, Jason A. Ross

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The formation of functional skeletal muscle is the consequence of both the differentiation and apoptosis of skeletal myoblasts. Ex vivo culture of skeletal myoblasts provides a tractable model for the study of these two coordinately regulated processes. We have previously reported that 23A2 myoblasts stably expressing a dominant negative Death Receptor 5 (A2:dnDR5 myoblasts) exhibit decreased basal mRNA and protein expression of the master muscle regulatory transcription factor MyoD. This decrease at the mRNA level is not a consequence of altered stability. Binding of the transcription factor SRF to a non-canonical CArG box within a serum response element (SRE) in …

Characterization Of The Immune Response To Anti-Mullerian Hormone, Justin M. Johnson

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Anti-Mullerian hormone (AMH) has been known since the mid-twentieth century as the substance secreted by the testes of the male fetus that is responsible for directing the proper development of the male reproductive organs from the primordial structures that would otherwise become the uterus and fallopian tubes. However, despite its longknown significance in male development, only recently has its importance in the adult female become evident. AMH is now considered to be not only a clinical indicator of the finite ovarian egg supply, but a vital regulator thereof throughout the reproductive life of the female. Dysfunctions in AMH regulation may …

Determinants Of Holliday Junction Formation And Resolution During Budding Yeast Meiosis, Marina Bykova

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In the process of meiosis, 4 haploid cells are produced from 1 diploid cell in two rounds of division. During prophase I of meiosis I, double strand breaks (DSBs) are created by the Spo11 protein, some of which are later repaired by homologous recombination into crossovers. During homologous recombination, structures called Holliday Junctions are formed, which later are resolved into crossovers to allow homologous chromosomes to separate. Ifthey are not resolved, the cell arrests and cannot progress further in meiosis. I have investigated three proteins known or suspected to be involved in Holliday Junction formation and resolution: the transcription factor …

Functional Ecology And Ecosystem Services Of Urban Trees, Milos Simovic

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Compared to their natural counterparts, trees in urban ecosystems experience distinctive environmental conditions which can be both beneficial and harmful to tree functions and fitness. Thus, the morphological, phenological, and physiological functions of trees in urban ecosystems can be unique and might not be predictable from patterns identified in natural forests where most research on tree ecology has occurred. To better understand how different tree species contribute to ecosystem services in urban environments, we estimated a number of key performance metrics and functional traits for species commonly planted in urban areas. Between April of 2017 and December of 2019, we …

Life Cycle And Morphological Characterization Of Colpodella Sp. (Atcc 50594) In Hay Medium, Troy A. Getty

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Colpodella species are free living biflagellated protists that prey on algae and bodonids in a process known as myzocytosis. Colpodella are phylogenetically closely related to Apicomplexa. Cyst and trophozoite stages have been identified in Colpodella species, however the mechanisms of encystation and excystation are unknown. The timing and transition of the life cycle stages are unknown. In the present study I investigated the life cycle of Colpodella sp. (ATCC 50594). Colpodella sp. were grown in diprotist cultures in tissue culture flasks containing Hay medium bacterized with Enterobacter aerogenes and containing Bodo caudatus as prey. I wanted to know the timing …

Role Of Ribosomal Protein Us9/Ys16 In Translation Initiation And Elongation In Yeast Saccharomyces Cerevisiae., Supriya Jindal

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The process of translation in all living cells is performed by ribosomes and is divided into four major steps (initiation, elongation, termination and ribosome recycling). Ribosomes consist of ribosomal RNA (rRNA) and proteins and are composed of two subunits small and large. There are three major sites for tRNA binding within ribosome: the (aminoacyl) A-site which accepts the aminoacyl (aa)-tRNA; the P-site, where the peptidyl-tRNA is formed and the (exit) E-site, where deacylated tRNA exits the ribosome. These sites are formed by both rRNA and ribosomal proteins. Though rRNA are involved in the catalysis of protein synthesis, the contribution of …

Functional Diversity And Restoration Of Meadows In Northeast Ohio, Keri Plevniak

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Restoration outcomes can be variable and there is a need to understand the short- and long-term responses of the vegetation community. It is important to see if management goals are being met by evaluating restoration outcomes. Restoration goals often include establishing native species, increasing conservation value, and providing pollinator resources throughout the year. Assessing communities is commonly done though a taxonomic approach by using species presence and abundance. Another method of evaluation is through functional traits or species traits and their abundance. Both give different perspectives on how the community is achieving management goals. While taxonomic assessments can give insight …

Investigation Of Ires-Mediated Translation Of Puma Mrna: Initiation Factor Requirements And Search For Itafs, Amra Ismail

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Translation initiation is the rate-limiting and tightly regulated step of protein synthesis. Cap-dependent translation initiation accounts for about 95% of cellular mRNAs. Around 3-5% of cellular mRNAs have been found to contain a cis-regulatory element (IRES) which can recruit ribosomes in a cap-independent manner. IRESs support protein synthesis under cellular stress conditions when cap-dependent translation is inhibited. Differentiation in 23A2 myoblast cells can be induced by culturing cells in serum-free differentiating media (DM). During 23A2 cellular myoblast differentiation, approximately 30% of cells undergo apoptosis as a result of stress caused by serum withdrawal in order to induce differentiation. The expression …

Biomimetic Macromolecules For Macrophage Targeting And Modulation, Joshua Whited

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Carbohydrate recognition has come to the forefront of biological aiming to uncover the mechanisms of physiological and pathological processes. Cell surface glycans are involved in processes including cellular adhesion, cell signaling, and immune response. A new approach for profiling cell surface glycans has great potential for a wide range of biomedical applications. Lectins have been conventionally used to determine the structure and function of glycoproteins, however, their numbers are still restricted compared to the number of glycan structures. Boronic acid has proven a remarkable small molecule capable of binding diols in aqueous solution. This interaction indicates boronic acid derived molecules …

A Synthesis Platform For Temperature Responsive Star Polymers, Schmitt J. Richard Jr.

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Star polymers are a class of branched polymers comprised of several polymer chains extending from a central point. Star polymers have applications in biopharmaceuticals where they have been proposed to be suitable drug delivery vehicles. Star polymers have traditionally been synthesized through chemical synthesis with added functionality provided by grafting on the arms. This complex synthesis can be simplified by using a biosynthetic approach which enables precise control of molecular weight and composition. This approach is demonstrated using star polymers with arms composed of a temperature responsive protein-based polymer termed elastin-like polypeptide (ELP). Star polymers are characterized based on the …

The Naked Mole Rat Possesses A Differently Phased Core Clock System Along With A Different Glucose Handling And Mtor Systems Compared To The Common Lab Mouse, Soumyaditya Ghosh

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INTRODUCTION The Naked Mole Rat [ Heterocephalus glaber] is a type of rodent -with the highest average lifespan-30 years-among all the rodent species. It lives underground in sealed burrow systems with little to no exposure to day-night transitions on the surface. It shows polyphasic round the clock activity within its subterranean burrow system. The activity and physiology of most animals are governed by the biological circadian clock. The biological circadian clock is a system that oscillates with a rough periodicity of 24 hours. Environmental zeitgebers regulate the entrainment of the circadian clock to the 24-hour day-night cycle and light is …

Stabilization Of Hypoxia Inducible Factor By Cobalt Chloride Can Alter Renal Transepithelial, Subhra Sankar Nag

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Kidney cyst expansion, stagnant fluid accumulation, and insufficient vascular supply can result in localized chronic ischemia-hypoxia in kidney cysts, as well as in normal renal epithelia adjacent to a cyst. We hypothesize that in normal epithelia near a cyst, the stabilization of Hypoxia Inducible Factor 1a (HIF1a), a major regulator of cellular response to hypoxia, can cause altered paracellular and transcellular transport, transforming a normal absorptive phenotype to a secretory and paracellularly leaky phenotype, leading to cyst expansion. Using 100 µmol/L cobalt chloride (CoCl2), HIF1a was stabilized in cellular nucleus of a mouse cortical collecting duct cell line (mCCD 1296 …

Cryptochromes Regulate Igf-1 Production And Signaling Through Control Of Jak2-Dependent Stat5b Phosphorylation, Amol Santosh Charudhari

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Insulin-like growth factor (IGF) signaling plays an important role in cell growth and proliferation and is implicated in regulation of cancer, metabolism, and aging. Here we report that IGF-1 level in blood and IGF-1 signaling demonstrates circadian rhythms. Circadian control occurs through cryptochromes (CRYs)—transcriptional repressors and components of the circadian clock. IGF-1 rhythms are disrupted in Cry-deficient mice, and IGF-1 level is reduced by 80% in these mice, which leads to reduced IGF signaling. In agreement, Cry-deficient mice have reduced body (~30% reduction) and organ size. Down-regulation of IGF-1 upon Cry deficiency correlates with reduced Igf-1 mRNA expression in the …

Restoration Of Black Oak (Quercus Velutina) Sand Barrens Via Three Different Habitat Management Approaches, David J. Kriska

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Disturbance regimes, i.e. frequent fires, historically maintained oak barrens until European settlement patterns, and eventually, Smoky the Bear and the fire suppression campaign of the U.S. Forest Service snuffed out the periodic flames. In the absence of a disturbance regime, ground layer floral composition at many historical oak sand barrens will change predominantly because of a buildup of leaf litter and shading of the soils. Termed mesophication, this process of ecological succession will drive Black Oak Sand Barrens to an alternate steady state. A survey conducted on Singer Lake Bog in Green, Ohio, demonstrated that succession shifted the community to …

Age And Sex-Specific Effect Of Caloric Restriction On Circadian Clock And Longevity-Associated Gene Expression, Arten Andreyevich Astafev

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The rhythms in the expression of circadian clock genes are affected by calorie restriction (CR), a dietary paradigm known to increase lifespan. In our current study, we show that circadian rhythms are influenced by sex and the effects of CR are different between males and females. In particular, we found a group of clock genes which showed a sex-dependent difference in expression, as well as in response to CR (Rev-Erb α, Ror γ and both Cryptochromes: Cry1 and Cry2 genes). Two clock genes showed no difference in expression but their response to CR showed sexual dimorphism (Ror α and Rev-Erb …

Sex-Bias In Lupus-Prone (Nzbxnzw)F1 Mice: The Interplay Of Gr1+Cd11b+ Cells, Testosterone And Genetics, Abhishek Trigunaite

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Systemic Lupus Erythematosus (SLE) is a prototypic systemic autoimmune disorder predominantly (9:1) targeting females. Currently, there is no cure for SLE and also there is a lack of disease predicting biomarkers. The major aim of my research is to determine the cellular basis of the sex bias in SLE and subsequently facilitate the development of better therapeutics and biomarkers for the disease. In (NZBxNZW) F1 mice a similar female bias is observed as in humans and testosterone is shown to be protective against lupus-like disease development, but the mechanism is not well understood.

We found higher levels of Gr1hiCD11b+ myeloid …

Calorie Restriction Effect On Circadian Clock Gene Expression, Sonal Arvind Patel

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Calorie Restriction (CR) is a powerful paradigm known to delay aging and thus increase longevity in several organisms, from yeast to non-human primates. Many molecular pathways have been proposed to mediate the beneficial effects of CR, however, the mechanism is still unknown. Circadian clock which is an internal time keeping system is regulated by feeding. Thus our aim was to study the effect of CR on the circadian clock. Here we show that CR significantly affects the expression of circadian clock genes in mice at the mRNA and protein levels, suggesting that CR reprograms the clocks at the transcriptional and …

Suppression Of Pulmonary Innate Immunity By Pneumoviruses, Jayeeta Dhar

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Pneumonia Virus of Mice (PVM) and Respiratory Syncytial Virus (RSV) are negative sense, single-stranded, enveloped RNA viruses from Pneumovirus genus, Paramyxoviridae family. RSV is the leading cause of respiratory diseases in infants. PVM causes similar respiratory illness in mice. PVM is used as an animal model to study RSV pathogenesis because of its similarity with RSV infection. Viral infection induces type I interferon (IFN) response as an antiviral strategy. PVM and RSV both have two non-structural (NS) proteins that are known to be IFN antagonists. While RSV can target different signaling components of IFN pathway, the mechanism of IFN suppression …

Effects Of Hypoxic Conditions On Skeletal Myoblasts, Shuai Zhao

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Hypoxic injury in skeletal muscle occurs in response to musculoskeletal traumas, diseases and following reconstructive and transplantation surgeries. However, the molecular mechanisms responsible for hypoxic injury in skeletal myoblasts have yet to be fully investigated. Understanding of these molecular mechanisms would identify potential therapeutic targets. Herein, we have determined that treatment with cobalt chloride (CoCl2, a hypoxia mimicking agent) first leads to an inhibition of proliferation followed by a decrease in the number of viable myoblasts over time. FACS analysis revealed a marked increase in G2/M arrest after 24 hours of treatment. We next assessed PARP cleavage, a well-known hallmark …

Rna-Protein Interactions In The U12-Dependent Spliceosome, Jagjit Singh

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Nuclear precursor messenger RNA (Pre-mRNA) splicing is an important regulatory step in metazoan gene expression. More than 99% of nuclear pre-mRNA introns are U2-type that are spliced by U2-dependent spliceosome containing U1, U2, U4, U5 and U6 snRNAs. Only less than 1% of the introns are U12-type and spliced by U11, U12, U4atac, U5 and U6atac snRNAs. U12 and U6atac snRNAs play a central role in the splicing of U12-dependent introns. Our previous work demonstrated that the conserved 3' stem-loop region of U6atac snRNA contains a U12-dependent spliceosome-specific targeting activity, however any potential molecular mechanism was unclear. We discovered that …

Ubiquitin-Proteasome System Modulates Platelet Function, Nilaksh Gupta

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Atherothrombotic diseases are responsible for more than 25 of all deaths worldwide. Anti-platelet drugs are the mainstay treatment because of the direct involvement of platelets in the initiation and propagation of thrombosis. However, the currently available anti-platelet drugs, such as antagonists of platelet receptors or of effector systems participating in platelet activation, have their own limitations. A new mode of affecting platelet reactivity may prove to offer unique advantages in a host of clinical settings. Proteasome inhibitors are in clinical use to treat hematologic cancers, but also reduce thrombosis. Whether the proteasome participates in platelet activation or function is opaque …

Acute And Chronic Rejection: Compartmentalization And Kinetics Ff Counterbalancing Signals In Cardiac Transplants, Anupurna Kaul

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Heart disease is the major cause of mortality in the United States and other parts of the world. Heart transplantation is the treatment of choice for patients with end stage heart failure. However, transplanted organs fail due to either acute or chronic rejection. This acute and chronic rejection impacts distinct compartments of cardiac allografts. Acute rejection is characterized by infiltration of mononuclear cells whereas chronic rejection is characterized by progressive narrowing of coronary arteries. In a minor histoincompatibility mismatch mouse model we found hearts transplanted from male to female C57BL/6 mice undergo an acute rejection with diffuse interstitial infiltrates at …

Circadian Regulation Of Mtor Signaling Via Bmal1 Dependent Mechanism, Rohini Vishal Khapre

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Understanding mechanisms of aging is important for the treatment and prevention of age-associated pathologies. However, these mechanisms are not well understood. Recently we have demonstrated that the circadian clock (an internal time keeping system) regulates longevity in mammals, but the molecular mechanisms are not known. The aim of our current study is to investigate a possible interconnection between the circadian clock and mTORC1 (mammalian target of Rapamycin) signaling pathway. mTORC1 pathway is a nutrient response pathway involved in many cellular processes many recent studies indicate a role of mTORC1 pathway in aging. Here we demonstrate that circadian system regulates mTORC1 …

Trypanosoma Brucei Telomere Functions In Antigenic Variation, Unnati M. Pandya

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Trypanosoma brucei is a protozoan parasite that causes sleeping sickness in humans and Nagana in cattle. They evade the host's immune defense by periodically switching their major surface antigen, variant surface glycoprotein (VSG), a phenomenon termed antigenic variation. Inside its mammalian host, bloodstream form (BF) T. brucei monoallelically expresses its major surface molecule VSG from the VSG Expression Sites (ESs) located at subtelomeric loci. Monoallelic VSG expression ensures effective antigenic variation and maximizes the efficiency of T. brucei pathogenesis. In the mid-gut of its insect host (tsetse), procyclic form (PF) T. brucei expresses procyclins as the major surface molecules and …

Study Of Role Of Ribosomal Protein L13a In Resolving Inflammation, Darshana Poddar

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Inflammation is an obligatory attempt of the host immune system to protect the body against infection. However, unregulated synthesis of pro-inflammatory products can have detrimental effects. Though mechanisms which contribute to inflammation are well appreciated, those that resolve inflammation are poorly understood. Therefore, understanding the molecular basis of such pathways will provide an entirely novel approach to treat and prevent inflammatory diseases. Transcript-selective translational control can regulate the expression of a set of inflammatory genes. We have identified one such mechanism in a novel animal model which relies on the abrogation of ribosomal protein L13a-dependent translational silencing by creating macrophage-specific …

Role Of Mtorc1 And Autophagy In The Regulation Of Chemoresistance In Leukemic B-Cells, Arishya Sharma

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Deregulated mTORC1 contributes to tumorigenesis and chemoresistance. However, mTORC1-specific inhibitors (rapalogs) show modest efficacy in the clinic, as they unleash the feedback inhibition on upstream, prosurvival pathways. An alternative approach is to target downstream functions of mTORC1. We investigated acquired resistance to fludarabine (Flu), a purine analog, active agent for chronic lymphocytic leukemia (CLL). Elevated phospho-p70S6k (T389), an mTORC1 activation marker, predicted Flu resistance in a panel of B-cell tumor lines, and primary CLL cells. However, the rapalogs induced moderate cell death in Flu-resistant (FluR) and primary CLL cells. Activation of oncogenic pathways, including mTORC1, induces profound metabolic reprogramming to …

Combinatorial Activation Of Stat3 By Egf And Thrombin In Endothelial Cells, Matthew S. Waitkus

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Endothelial cells line the luminal surface of blood vessels and form a regulatory interface between the bloodstream and underlying tissues. The endothelium responds to diverse, and potentially conflicting, environmental signals to regulate vessel growth, leukocyte adhesion, thrombogenicity, and vascular tone. Signaling pathways may interact, or "crosstalk," in combinatorial signaling environments to enable cells to process disparate extracellular information at downstream signaling nodes and formulate appropriate biological responses based on combinations of extracellular stimuli. We have reported that simultaneous stimulation of endothelial cells with EGF and thrombin synergistically induces expression of immediate early genes (IEGs) associated with growth and angiogenesis. We …