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- Innate immunity (4)
- Degranulation (3)
- Midline (3)
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- Differentiation (2)
- Exocytosis (2)
- MESCs (2)
- Mast cell (2)
- Mast cells (2)
- MiRNA (2)
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- Mirtron (2)
- MsaABCR operon (2)
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- Staphylococcus aureus (2)
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- Alpha-gal (1)
- Annelid (1)
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- Antibody (1)
- Antiviral (1)
- Bab2 (1)
- Bioimaging (1)
- Bric-a-brac (1)
- C-Jun-N-terminal kinase (1)
- CG17386 (1)
- Cell signaling (1)
- CoA (1)
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Uncovering Novel Small Regulatory Rna In Protostome, Sweta Khanal
Uncovering Novel Small Regulatory Rna In Protostome, Sweta Khanal
Dissertations
Small RNAs play pivotal roles in post-transcriptional gene regulation across diverse phylum of protostomes. In this study, we investigate the functional significance of atypical miRNAs, mirtron miR-1017 in Drosophila. Through ectopic expression in neuronal cells, we demonstrate that miR-1017 extends lifespan by targeting its host transcript, acetylcholine receptor Dα2, and influencing its splicing. This novel trans-regulatory function suggests a mechanism for mirtron evolution, highlighting the interplay between splicing and post-transcriptional regulation. Additionally, we profile small RNA populations in the polychaete developmental model Capitella teleta, shedding light on the small RNA landscape in annelid worms. Our analysis reveals a rich …
Uptake And Localization Of Poly(2-Vinyl-4,4-Dimethyl Azlactone) Modified With Rhodamine- And Coumarin-Based Molecules In Human Embryonic Kidney (Hek293) Cells, Garrett Tassin
Honors Theses
Fluorescence microscopy is a bioimaging technology that utilizes the excitation and emission of fluorophores to identify cellular structures, processes, and interactive events. Natural and synthetic organic dyes are frequently used in fluorescence microscopy techniques for imaging, therapeutic, and biomedical applications. It is also employed in the development of novel organic dyes for innovative methods of drug delivery and labelling. In this study, we investigate the behavior of an organic dye consisting of the post-polymerization modification of poly(2-vinyl-4,4-dimethyl azlactone) (PVDMA) with tetramethyl rhodamine cadaverine (TMR) and a coumarin-based molecule (DBAC). Using cultured HEK293 cells and fluorescence confocal microscopy, we show the …
Delineating The Exocytic Fusion Machinery Required For Mast Cell Exocytosis, Pratikshya Adhikari
Delineating The Exocytic Fusion Machinery Required For Mast Cell Exocytosis, Pratikshya Adhikari
Dissertations
Mast cells undergo exocytosis to release a wide array of inflammatory mediators by utilizing membrane fusion proteins-SNAREs (soluble-N-ethyl-maleimide sensitive factor attachment protein receptor) along with essential regulatory Munc18 and Munc13 proteins. Accumulating evidence in mast cell biology suggests the existence of distinct pools of mast cell mediators. However, the precise mechanism underlying the release of each mast cell mediator is not clear. To determine whether different exocytic machineries are required for differential mediator release, I used reconstitution to investigate the differential role of Munc18s in fusion machinery regulation. Munc18a and Munc18c stimulated VAMP2 and VAMP3-mediated lipid mixing, whereas Munc18b only …
Dicer Represses Antiviral Innate Immunity Pathways In Mouse Embryonic Stem Cells, Chandan Gurung
Dicer Represses Antiviral Innate Immunity Pathways In Mouse Embryonic Stem Cells, Chandan Gurung
Dissertations
Recent studies have demonstrated that embryonic stem cells (ESCs) are deficient in expressing type I interferons (IFN), the cytokines that play key roles in antiviral responses. However, the underlying molecular mechanisms and biological implications of this finding are poorly understood. In this study, I used a synthetic RNA-based assay that can simultaneously assess multiple forms of antiviral responses in ESCs. Dicer is an enzyme essential for RNA interference (RNAi), which is used as a major antiviral mechanism in invertebrates but not clear in vertebrates. RNAi activity is detected in wild-type ESCs but is abolished in Dicer knockout ESCs (D−/−ESCs) as …
Human 5’-Tailed Mirtrons Are Processed By Rnasep, Mohammad Farid Zia
Human 5’-Tailed Mirtrons Are Processed By Rnasep, Mohammad Farid Zia
Dissertations
Approximately a thousand microRNAs (miRNAs) are documented from human cells. A third appear to transit non-canonical pathways that typically bypass processing by Drosha, the dedicated nuclear miRNA producing enzyme. The largest class of non-canonical miRNAs are mirtrons which eschew Drosha to mature through spliceosome activity. While mirtrons are found in several configurations, the vast majority of human mirtron species are 5’-tailed. For these mirtrons, a 3’ splice site defines the 3’ end of their hairpin precursor while a “tail” of variable length separates the 5’ base of the hairpin from the nearest splice site. How this tail is removed is …
Molecular Characterization Of Galectin From Amblyomma Americanum In Context Of Α-Gal Syndrome, Sumar Beauti
Molecular Characterization Of Galectin From Amblyomma Americanum In Context Of Α-Gal Syndrome, Sumar Beauti
Honors Theses
The lone star tick Amblyomma americanum is a vector of various disease-causing pathogens and tick-borne alpha-gal syndrome (AGS) with rapidly expanding populations in the south- and northeast regions of the United States. This study aimed to molecularly characterize galectin and determine its involvement in galactose-α-1,3-galactose (α-gal) synthesis, transport, reproductive fitness, and microbial homeostasis in this tick. The lone star tick galectin possesses two conserved carbohydrate recognition domains and shares homology with other Ixodid tick galectins. Time and tissue-dependent expression data shows that galectin is constantly expressed in salivary glands, midgut, and ovary tissues. An RNA interference approach was used to …
Characterizing Erad And Antioxidant Response In Ixodes Scapularis Ise6 Cells During Borrelia Infection, Latoyia Downs
Characterizing Erad And Antioxidant Response In Ixodes Scapularis Ise6 Cells During Borrelia Infection, Latoyia Downs
Master's Theses
Ticks are hematophagous ectoparasites that transmit a multitude of diseases to humans. Borrelia burgdorferi (BB) and Borrelia miyamotoi (BM) are both tick-borne pathogens that cause disease in humans and are transmitted by the black-legged tick (Ixodes scapularis). A byproduct of blood digestion generates reactive oxygen species that are toxic and cause oxidative stress which promotes cellular damage and dysfunction. The endoplasmic reticulum (ER) is especially affected by oxidative stress, resulting in a buildup of improperly folded proteins in the ER lumen called ER stress. To prevent cellular damage, the tick utilizes an antioxidant system to neutralize ROS and mechanisms …
Evaluating Biogenesis Of 5’-Tailed Mirtrons, Jonathan D. Hoover
Evaluating Biogenesis Of 5’-Tailed Mirtrons, Jonathan D. Hoover
Honors Theses
MicroRNAs are 22 nucleotide, non-coding RNAs that serve as substrates for Argonaute proteins to induce RNA interference pathways. Intron-derived miRNA precursors called “mirtrons” have been identified and classified primarily through deep sequencing methods. Unlike most miRNAs, mirtrons are derived from splicing events and also exhibit high levels of post-transcriptional nucleotide addition to hairpin precursors. Most relevant among these modifications is 3’ uridylation as it inhibits mirtron biogenesis in multiple model systems. Mirtrons may also possess additional nucleotides adjacent to the pre-miRNA hairpin at the 3’ and/or 5’ ends. These nucleotide “tails” are removed prior to Dicer cleaving the hairpin. In …
Investigating Roles For Rna Turnover Processes In Cell Signaling Through Drosophila Melanogaster Genetic Mosaics, Sudiksha Rathan Kumar
Investigating Roles For Rna Turnover Processes In Cell Signaling Through Drosophila Melanogaster Genetic Mosaics, Sudiksha Rathan Kumar
Honors Theses
The process of cell signaling is vital in organisms for proper development as it determines the fate and function of cells. This process is highly regulated by myriad interactions between signaling pathway components and gene expression mechanisms. RNA turnover is a type of RNA processing that degrades RNA. It plays an essential role in homeostasis and environmental changes; however, its influence on signaling pathways is currently unknown. In the present study, the effect of RNA turnover processes on cell signaling was analyzed using the genetic mosaics in Drosophila melanogaster. Fly mosaics were created by breeding RNAi lines targeting cellular …
Mouse Embryonic Stem Cells Are Sensitive To The Cytotoxicity Of Nitric Oxide: Biological Implications For Early Embryogenesis, Mckenzie C. Hargis
Mouse Embryonic Stem Cells Are Sensitive To The Cytotoxicity Of Nitric Oxide: Biological Implications For Early Embryogenesis, Mckenzie C. Hargis
Honors Theses
The early embryo, before implantation, is at a very vulnerable stage in development where it faces various inflammatory cytokines throughout the implantation process. In this stage, the cells in the blastocyst, the preimplantation stage embryo, must proliferate rapidly for tissue formation. However, it is known that inflammatory cytokines can inhibit cell proliferation. Previous studies have shown that embryonic stem cells (mESCs), the major cell component in the blastocyst, are unresponsive to treatments of tumor necrosis factor ⍺ (TNF⍺) and interferon 𝛾 (IFN𝛾), two inflammatory cytokines involved in the implantation process. Treatment of mESC-differentiated fibroblasts (mESC-FBs) with TNF⍺ and IFN𝛾 in …
The Histoplasma Capsulatum Ddr48 Gene Is Required For Survival Within Macrophages And Resistance To Oxidative Stress And Antifungal Drugs, Logan Blancett
The Histoplasma Capsulatum Ddr48 Gene Is Required For Survival Within Macrophages And Resistance To Oxidative Stress And Antifungal Drugs, Logan Blancett
Dissertations
Histoplasma capsulatum(Hc)is a systemic, dimorphic fungal pathogen that affects upwards of 500,000 individuals in the United States annually. Hc grows as a multicellular mold at environmental temperatures; whereas, upon inhalation into a human or other mammalian host, it transforms into a unicellular, pathogenic yeast. The research presented in this dissertation is focused on characterizing the DNA damage-responsive gene HcDDR48. HcDDR48was originally isolated via a subtractive DNA library enriched for transcripts enriched in the mold-phase of Hcgrowth. Upon further analysis we found that HcDDR48is not just expressed in the mold morphotype, but both growth programs …
The Antimicrobial Activity And Cellular Targets Of Plant Derived Aldehydes And Degradable Pro-Antimicrobial Networks In Pseudomonas Aeruginosa, Yetunde Adewunmi
The Antimicrobial Activity And Cellular Targets Of Plant Derived Aldehydes And Degradable Pro-Antimicrobial Networks In Pseudomonas Aeruginosa, Yetunde Adewunmi
Dissertations
Essential oils (EOs) are plant-derived products that have been long exploited for their antimicrobial activities in medicine, agriculture, and food preservation. EOs represent a promising alternative to conventional antibiotics due to the broad-range antimicrobial activity, low toxicity to human commensal bacteria, and the capacity to kill microorganisms without promoting resistance. Despite the progress in the understanding of the biological activity of EOs, many aspects of their mode of action remain inconclusive. The overarching aim of this work was to address these gaps by studying molecular interactions between antimicrobial plant aldehydes and the opportunistic human pathogen Pseudomonas aeruginosa. We initiated …
Attenuated Responses To Inflammatory Cytokines In Mouse Embryonic Stem Cells: Biological Implications And The Molecular Basis, Bohan Chen
Master's Theses
Embryonic stem cells (ESCs) have attracted intense interest due to their great potential for regenerative medicine. However, their immune property is an overlooked but a significant issue that needs to be thoroughly investigated not only to resolve the concern for therapeutic applications but also for further understanding the early stage of organismal development. Recent studies demonstrated that ESCs are deficient in innate immune responses to viral/bacterial infections and inflammatory cytokines. Inflammatory conditions generally inhibit cell proliferation, which could be detrimental to ESCs, since cell proliferation is their dedicated task during early embryogenesis. Thus, I hypothesize that the attenuated innate immunity …
The Subcloning And Expression Of Munc18a In Escherichia Coli For Antibody Production And Analysis In Mast Cell Degranulation Reactions, Brandi Goble
Honors Theses
Mast cells are secretory cells responsible for fighting off infection through the early recognition of pathogens. This process is completed through the secretion of proinflammatory mediators that are stored in secretory granules within the cytoplasm of the cell. The degranulation secretion process relies on regulated fusion of secretory granules to the cell membrane via membrane-bound SNARE proteins that bridges the two opposed membranes. The intricate regulation of SNARE-mediated mast cell degranulation is not well understood. However, Sec1/Munc18 (SM) proteins, specifically the Munc18 isoforms, are known to play a critical role in the process (Brochetta, et. al., 2014). The Xu lab …
Development Of Innate Immunity During In Vitro Differentiation Of Mouse Embryonic Stem Cells, William D'Angelo
Development Of Innate Immunity During In Vitro Differentiation Of Mouse Embryonic Stem Cells, William D'Angelo
Dissertations
Embryonic stem cells (ESCs) hold enormous promise for the goals of regenerative medicine and tissue engineering, and extraordinary progress has been made in defining conditions for differentiation to desired cell types. However, an often overlooked aspect of ESC biology is innate immunity, the ability of cells to detect and respond to pathogens and inflammatory cytokines. A number of recent studies by our lab and others have established that ESCs and other types of pluripotent cells from both mice and humans do not mount typical immune responses to viral or bacterial stimuli. There are also indications that various cell types differentiated …
Uncovering The Identity And Metabolism Of Bacterial Coa-Rna, Joseph R. Spangler
Uncovering The Identity And Metabolism Of Bacterial Coa-Rna, Joseph R. Spangler
Dissertations
Coenzyme A is an indispensable molecule in all known life with roles in metabolism, gene regulation, and macromolecule synthesis. As CoA is derived from RNA itself, it’s incorporation into RNA by in vitro methods has proven useful in research probing the origin of life based on the RNA World theory. The discovery in contemporary bacteria of RNA modified with CoA, however, provided an unexpected twist to previously well-characterized bacterial systems. The identity of sequences associated with CoA-RNA has been elusive since their discovery in 2009 based on the difficulties in isolation while maintaining RNA quality. The aim of this study …
Reconstitution Of Munc18-Dependent Mast Cell Degranulation, Matthew Grant Arnold
Reconstitution Of Munc18-Dependent Mast Cell Degranulation, Matthew Grant Arnold
Master's Theses
Mast cells are specialized secretory leukocytes that play diverse roles in the body, mediated by release of histamine and other pharmacologically active substances. Although offering essential protection in innate and adaptive immunity, mast cells are also essential to the progression of inflammatory diseases, including allergy and asthma, (Theoharides et al., 2012). Exocytosis of pro-inflammatory mast cell mediators in response to otherwise innocuous antigens relies on evolutionarily-conserved membrane fusion machinery. These proteins, called SNAREs are regulated by the Sec1/Munc18 (SM) protein family (Axle Lorentz, Baumann, Vitte, & Blank, 2012). Mast cells express three mammalian Munc18 isoforms (a, b, and c), which …
The Uas-Gal4 System In D. Melanogaster: An Insight Into The Influence Of Micrornas On The Developmental Pathways Of The Wing, Emily R. Wilson
The Uas-Gal4 System In D. Melanogaster: An Insight Into The Influence Of Micrornas On The Developmental Pathways Of The Wing, Emily R. Wilson
Honors Theses
By examining genetic pathways in D. melanogaster, a better understanding of the homologous regulatory mechanisms in humans can be utilized to further enhance knowledge of the roles of microRNA within development. This study utilizes the UAS-Gal4 system in order to produce a mutant phenotype capable of being visually studied and analyzed, focusing on the developmental pathway of the wing in D. melanogaster. Dissections of the wandering third instar larvae yielded wing disc tissue expressing the downregulation of loquacious and CG17386.
Characterization Of Embryonic Stem Cell-Differentiated Cells As Mesenchymal Stem Cells, Rachael N. Kuehn
Characterization Of Embryonic Stem Cell-Differentiated Cells As Mesenchymal Stem Cells, Rachael N. Kuehn
Honors Theses
Embryonic stem cells (ESCs), due to their ability to differentiate into different cell types while still maintaining a high proliferation capacity, have been considered as a potential cell source in regenerative medicine. However, current ESC differentiation methods are low yielding and create heterogeneous cell populations. If transplanted in the human body, differentiated ESCs could be rejected by the immune system, form tumors, or may not function normally within the human body. On the other hand, mesenchymal stem cells (MSCs), a type of adult stem cell typically derived from bone marrow, have proved to be excellent candidates in clinical applications due …
Deciphering The Functional Collaboration Of Mid And Bric-A-Brac 2 As Potential Regulators Of Cellular Proliferation Within Adult Drosophila Ovaries, Petra Visic
Master's Theses
Stem cell niches are highly organized and specialized microenvironments located within specific tissues of both vertebrate and invertebrate organisms [1]. In Drosophila melanogaster, three distinct stem cell niches have been identified within the ovary including the germline stem cell (GSC), follicle stem cell (FSC), and escort stem cell (ESC) niche. Recently, Fregoso-Lomas et al. [2] reported that Gurken/Epidermal Growth Factor Receptor (EGFR) signaling is modulated within posterior ovarian follicle cells by Midline (Mid). The mid gene encodes a T-box transcription factor protein that specifies cell fates in the developing heart [3][4], central nervous system [5][6], epidermis [7], and eye …
Activation Of Nf-Κb Transcription Factor During In Vitro Differentiation Of Mouse Embryonic Stem Cells, Natalya A. Ortolano
Activation Of Nf-Κb Transcription Factor During In Vitro Differentiation Of Mouse Embryonic Stem Cells, Natalya A. Ortolano
Honors Theses
Embryonic stem cells (ESCs) are a promising cell source for regenerative medicine. However, recent studies indicated that ESCs and ESC-derived cells (ESC-DCs) lack functional innate immunity against various pathogens and inflammatory cytokines. This presents a barrier to clinical application, as ESC-DCs would be placed in a wound site and exposed to pathogens and inflammatory cytokines. Using mouse ESCs (mESCs) as a model, we recently demonstrated that they are deficient in expressing type I interferons (IFN) and inflammatory cytokines. To determine the molecular basis for this finding, this study examined the activation state of nuclear factor-κB (NF-κB), a transcription factor that …
The Drosophila T-Box Transcription Factor Midline Functions Within The Insulin/Akt And C-Jun-N-Terminal Kinase Signaling Pathways To Regulate Interomatidial Bristle Formation And Cell Survival, Qichuan Chen
Master's Theses
From a genetic and allelic modifier screen, we report that the Drosophila melanogaster T-box transcription factor midline (mid), a homolog to the human TBX20 gene, interacts with dFOXO within the insulin receptor (InR) and the c-Jun-N-terminal kinase (JNK) signaling pathways to regulate interommatidial bristle (IOB) formation. Previous studies have identified mid’s role in cell fate specification of sensory organ precursor cells in conjunction with the Notch-Delta signaling pathway (Das et al., 2013). The Notch, InR, and JNK signaling pathways regulate dFOXO activity under conditions of stress. Thus, we determined the effects of oxidative stress and metabolic stress by …
Identification And Characterization Of Msab Gene Involved In Biofilm Formation And Virulence In Staphylococcus Aureus, Amelsaad Elbarasi
Identification And Characterization Of Msab Gene Involved In Biofilm Formation And Virulence In Staphylococcus Aureus, Amelsaad Elbarasi
Master's Theses
Staphylococcus aureus is an important human pathogen that causes a wide variety of life-threatening infections ranging from minor skin and oral infections to severe infections, such as bacteremia, pneumonia, osteomyelitis, or endocarditis due to the presence and secretion of a large number of virulence factors that are controlled by global virulence regulators in complex networks. Furthermore, S. aureus infections have become a threat to public health because of their high potential to form biofilm, and their ability to resist a wide range of antibiotics has exacerbated further. Therefore, understanding the regulatory networks and developing a drug targeting these networks has …
Role Of Msaa Gene In Regulation Of The Msaabcr Operon And Biofilm Development In Staphylococcus Aureus, Ahmed Alzuway
Role Of Msaa Gene In Regulation Of The Msaabcr Operon And Biofilm Development In Staphylococcus Aureus, Ahmed Alzuway
Master's Theses
Staphylococcus aureus is an important human pathogen that causes wide variety of diseases ranging from chronic biofilm associated infection to acute life threatening infection such as bacteremia, pneumonia, osteomyelitis, or endocarditis, despite the progress with antibiotics used in the treatment of bacterial infections. Furthermore, increased use of prosthetic and indwelling devices in modern medical practices has led to increased infections due to S. aureus. Treating S. aureus infections have become difficult owing to its ability to resist most of the antibiotics; this problem is further exacerbated by ability of MRSA strains to form biofilms. Emergence of community-acquired methicillin resistance …
Antiviral Responses In Mouse Embryonic Stem Cells: Differential Development Of Cellular Mechanisms In Type I Interferon Production And Response, Ruoxing Wang
Dissertations
Embryonic stem cells (ESCs) have been recognized as a promising cell source for regenerative medicine. Intensive research over the past decade has led to the possibility that ESC-derived cells will be used for the treatment of human diseases. However, increasing evidence indicates that ESC-derived cells generated by the current differentiation methods are not fully functional. It is recently recognized that ESC-derived cells lack innate immunity to a wide range of infectious agents and inflammatory cytokines. When used in patients, ESC-derived cells would be placed in wounded sites that are exposed to various pathogens and inflammatory cytokines; therefore, their viability and …
Subcloning And Expression Of Complexin Isoforms Involved In Mast Cell Degranulation, Cameron Blake King
Subcloning And Expression Of Complexin Isoforms Involved In Mast Cell Degranulation, Cameron Blake King
Honors Theses
Mast cells play an important role in the immune system by releasing chemicals such as chemokines and cytokines once they are stimulated. These products are released after stimulation by a process called mast cell degranulation. Mast cell degranulation is accomplished when vesicles containing the chemicals inside the mast cell fuse with the mast cell membrane via SNARE-mediated (Soluble NSF Attachment Protein Receptors) membrane fusion. This family of proteins consists of syntaxin, SNAP 25-like protein, and synaptobrevin/VAMP (Vesicle Associated Membrane Protein)(2). Comlexin isoforms (complexin 1,2,3,and 4) have been known to regulate this system in a fashion that is still unclear. In …
Investigating The Co-Regulatory Role Of Midline And Extramacrochaetae In Regulating Eye Development And Vision In Drosophila Melanogaster, Lillian M. Forstall
Investigating The Co-Regulatory Role Of Midline And Extramacrochaetae In Regulating Eye Development And Vision In Drosophila Melanogaster, Lillian M. Forstall
Honors Theses
The Honors thesis research focused on the roles of extramacrochaetae and midline in regulating eye development and the vision of Drosophila melanogaster. It is known from previous studies that extramacrochaetae (emc) and midline (mid) independently regulate the formation of ommatidial units in the Drosophila compound eye. However, the thesis focuses on the interaction of these two genes and their co-dependent roles in regulating eye development. This study also attempts to explain the recovered formation of ommatidial units and interommatidial bristles when the expression of both of these genes is reduced and whether flies doubly mutant …
Developmental Expression Profile Of Bric Á Brac 2, Midline And H15 In The Developing Eye And Central Nervous System Of Drosophila Melanogaster, Petra Visic
Honors Theses
Aberrant activity of a single gene can lead towards development of cancerous cells. Drosophila melanogaster is a useful model system to study cancer because there is high degree of evolutionary conservation in signaling pathways between humans and flies that play major roles in regulating cell proliferation and growth (Miles et al., 2011). At The University of Southern Mississippi (USM), Dr. Leal’s lab has gathered evidence suggesting that bab1 and bab2 interact with the T-box gene midline (mid) and its paralog H15, while the early developmental function of bab1 and bab2 remains unknown. That is why elucidating the early …