Cell Biology Commons^™

Serpinb1 Effects On Human Pancreatic Ductal Cells, Praneeth Goli

College of Arts & Sciences Senior Theses

Rebuilding or expanding the body’s ability to regulate blood glucose levels by restoring or increasing the body’s ability to produce insulin, long-term, would be a major advancement in diabetes (type-1 and type-2) treatment and could serve as a potential cure [1]. Therefore, the holy grail of diabetes treatment and cure research is to regenerate or increase the insulin producing cells in diabetic individuals (type-1 and type-2) [1, 2]. SerpinB1 has been implicated as a biomolecular signal that stimulates an increase in insulin cell mass in murine and zebrafish models that exhibit physiologically normal compensatory islet hyperplasia. In humans, SerpinB1 is …

Structural And Functional Characterization Of Mitochondria With Trna Mutations, Brooke Henry

Biological Sciences Undergraduate Honors Theses

Mitochondrial encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) is one of the most common disorders associated with mitochondrial tRNA mutations. One of the most common causes of MELAS is mutation in the MT-TL1 gene which codes for mitochondrial tRNA Leucine (UUR). Mutation in MT-TE gene, another mitochondrial gene which encodes for mitochondrial tRNA Glutamate (GAA/G), has been implicated in various mitochondrial related myopathies. It remains unclear how point mutations in these tRNA genes result in disease onset and progression. Here, we report an early comparative analysis of fibroblast cell lines derived from patients carrying two different tRNA mutations: m.3243A>G …

Photoactivatable Cre Recombinase 3.0 For In Vivo Mouse Applications, Kumi Morikawa, Kazuhiro Furuhashi, Carmen De Sena-Tomas, Alvaro L. Garcia-Garcia, Ramsey Bekdash, Alison D. Klein, Nicholas Gallerani, Hannah E. Yamamoto, Seon Hye E. Park, Grant S. Collins, Fuun Kawano, Moritoshi Sato, Chyuan Sheng Lin, Kimara L. Targoff, Edmund Au, Michael C. Salling, Masayuki Yazawa

School of Medicine Faculty Publications

Optogenetic genome engineering tools enable spatiotemporal control of gene expression and provide new insight into biological function. Here, we report the new version of genetically encoded photoactivatable (PA) Cre recombinase, PA-Cre 3.0. To improve PA-Cre technology, we compare light-dimerization tools and optimize for mammalian expression using a CAG promoter, Magnets, and 2A self-cleaving peptide. To prevent background recombination caused by the high sequence similarity in the dimerization domains, we modify the codons for mouse gene targeting and viral production. Overall, these modifications significantly reduce dark leak activity and improve blue-light induction developing our new version, PA-Cre 3.0. As a resource, …

Assessing The Influence Of Riparian Invasion By The Shrub Lonicera Maackii On Terrestrial Subsidies To Headwater Streams, Erin C. Rowekamp, Julia I. Chapman, Ryan Mcewan

Biology Faculty Publications

Invasion of Amur honeysuckle (Lonicera maackii) into riparian areas of headwater streams creates strong potential for alterations of terrestrial subsidies that may drive bottom-up effects on aquatic biota. In this study, we analyzed effects of L. maackii on terrestrial subsidies in stream sites that represented a gradient of invasion intensity in temperate deciduous forests of southwestern Ohio (USA). Leaf litter biomass, terrestrial and aquatic fine woody debris (0.5–9.9 cm diameter) volume and count, and terrestrial and aquatic coarse woody debris (>9.9 cm diameter) volume were analyzed. Based on previous research, we hypothesized that sites with a higher …

A Putative Cystathionine Beta-Synthase Homolog Of Mycolicibacterium Smegmatis Is Involved In De Novo Cysteine Biosynthesis, Saroj Kumar Mahato

Graduate Theses and Dissertations

Mycobacteria include serious pathogens of humans and animals. Mycolicibacterium smegmatis is a non-pathogenic model that is widely used to study core mycobacterial metabolism. This thesis explores mycobacterial pathways of cysteine biosynthesis by generating and study of genetic mutants of M. smegmatis. Published in vitro biochemical studies had revealed three independent routes to cysteine synthesis in mycobacteria involving separate homologs of cysteine synthase, namely CysK1, CysK2, and CysM. However, in vivo data were lacking. The M. smegmatis genome encodes only a CysM homolog and lacks orthologs for CysK1 or CysK2. The gene that codes for CysM is a part of an …

Cloning, Protein Expression, And Characterization Of Interleukin 1 Alpha, Musaab Habeeb Ali Al Ameer

Graduate Theses and Dissertations

Recombinant DNA technology and the ability to produce recombinant proteins have significantly changed the world of pharmaceutical market. Recombinant DNA technology using E. coli cells has successfully produced many therapeutic proteins. In this study, the designed tag Ark-RUBY-tag facilitates rapid, and reproducible purification of recombinant proteins expressed as inclusion bodies in E. coli cells. Purification of Ark-RUBY-fused recombinant protein(s) can be obtained by using imidazole fractions. Target protein can be easily removed from the Ark-RUBY-tag by enzymatic cleavage. Ark-RUBY-fused recombinant proteins can be quantitatively detected using polyclonal antibodies. Ark-RUBY-tag can be used to purify small peptides. By using Ark-RUBY-tag, purified …

Development Of An Auto-Bioluminescent Lung Cell Line For Evaluation Of In Vitro Effects Of E-Cigarette Compounds On Normal Cell Proliferation, Olivia Morrison

Honors Theses

The effects of smoking have long been studied; however, with the rise of electronic cigarettes there is a growing interest in the effects of this smoking substitute. Marketed as a safer alternative to tobacco cigarettes, the popularity of electronic cigarette (e-cigarette) use has dramatically increased. This has demanded regulatory methods to be put into effect, however, scientific study is only beginning on the new fad to determine the impacts of its use on the human body. The present study aims to examine the biological effects of lung cell exposure to tobacco alkaloids found in electronic cigarette filling solutions. We hypothesize …

Mitochondrial Deletions And Their Disease-Causing Effects, Austin Bell

Biological Sciences Undergraduate Honors Theses

The mitochondria perform a plethora of important functions within the cell, with one of the most paramount being ATP production. Deregulation of its function can have dire consequences on cellular functions. Mutations such as deletions within the mitochondrial DNA (mtDNA) can cause disease within the patients affected. These diseases often affect children, causing symptoms such as gradual loss of eyesight and hearing, diabetes, and other problems that lower the quality of life. The mitochondria are also very dynamic organelles that undergo rounds of fission and fusion to keep up with the metabolic needs of cells, necessitating a homeostatic balance between …

Sensing Ribonuclease H Activity With Dna Nanoswitches, Ruju Trivedi

Biological Sciences

Ribonuclease H (RNase H) is a damage-repair protein and ribonuclease that specifically catalyzes the hydrolysis of RNA in an RNA/DNA duplex and breaks down RNA/DNA junctions. It plays an important role in a variety of biological processes including DNA replication, DNA repair, and transcription. It is also pivotal in anti-HIV drug development and the analysis of cellular processes and has been shown to be a potential therapeutic target for various neoplastic diseases. This thesis discusses a unique assay based on DNA nanoswitches to detect RNase H levels and activity. The assay is based on conformational changes of DNA nanoswitches in …

Characterization Of A Mycoplasma Pneumoniae Cards Toxin Mutant, Nikaash Pasnoori

Honors Scholar Theses

Mycoplasma pneumoniae is a high-burden pathogen which causes mild to significant infections of the respiratory system. According to the CDC, an estimated two million cases occur yearly in the United States alone, demonstrating the widespread effect of the pathogen. In addition to being the cause of respiratory infections, M. pneumoniae has also been implicated in exacerbating pre-existing asthma conditions. These morbidities make finding a vaccine candidate a vital part of easing the healthcare burden caused by the pathogen. The current mechanism of infection is unknown, but recent evidence points to the Community Acquired Respiratory Distress Syndrome (CARDS) toxin as being …

Heterogeneous Nuclear Ribonucleoprotein K (Hnrnp K) Overexpression And Its Interaction With Runx1 Rna In Acute Myeloid Leukemia, Marisa Aitken

Dissertations and Theses (Open Access)

Acute myeloid leukemia (AML) is an often devastating hematologic malignancy with 5-year overall survival lingering near 20%. Acquiring a deeper understanding of molecular underpinnings of leukemogenesis will provide a basis for developing more effective therapeutic strategies for patients with AML.

Here, we identified overexpression of hnRNP K as a recurrent abnormality in a subset (~20%) of AML patients. High levels of this RNA-binding protein associated with inferior clinical outcomes in de novo AML. Thus, to evaluate its putative oncogenic capacity in myeloid disease, we overexpressed hnRNP K in murine hematopoietic stem and progenitor cells isolated from fetal liver cells (FLCs). …

Understanding Intercellular Signaling During Lung Injury-Repair, Margo Patricia Cain

Dissertations and Theses (Open Access)

The mammalian lung is a complex system of specialized cell types with precise spatial organization designed to cooperate to perform gas exchange. These cell types together coordinate organ development, homeostasis, and repair after injury through signals either presented or secreted, known as ligands, to be received by receptors on the surface of another, or in some cases, the same cell. The alveolar region of the lung, the primary region of gas exchange, responds to various types of injuries with different lung repair mechanisms. In order to explore how the various cell types in the lung communicate to drive tissue repair …

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 …

A Novel Uv Resistance In Rad23-Depleted Tetrahymena Thermophila, Emily M. Schmoll

Graduate Theses/Dissertations

Rad23 is a highly conserved cellular scaffold protein which participates in the nucleotide excision repair pathway and ubiquitin proteasome system. It is hypothesized that the contradictory roles of Rad23 within these two systems, acting to enhance stability or facilitate degradation respectively, could be regulated via post-translational modification of the ubiquitin-like domain of the protein. To this end, a Rad23 somatic knockout cell line was established in Tetrahymena thermophila, with the eventual goal of knocking in a mutant Rad23 protein lacking potential for UbL ubiquitylation. In contrast to the UV-sensitive phenotype observed in similar models, Rad23-depleted Tetrahymena cell lines displayed significantly …

Myosin V-Mediated Cargo Traffic Toward The Trans-Golgi Network And Myosin V Implications In Snc1 Exocytosis, Vy Ngoc Khanh Nguyen

Graduate Theses/Dissertations

Retrieval of cargo proteins from the endosome towards the trans-Golgi network (TGN) is a crucial intracellular process for cellular homeostasis. Its dysfunction is associated with pathogenesis of Alzheimer and Parkinson's diseases. Myosin family proteins are cellular motors walking along actin filaments by utilizing the chemical energy from ATP hydrolysis, known to involve in pleiotropic cellular trafficking pathways. However, the question of whether myosins play a role in the trafficking of Snc1 and Vps10 has not been addressed yet. The present study assesses the potential roles of all five yeast myosins in the recycling of two membrane cargo, Snc1 and …

Development Of In Vitro Models To Study The Rapid Extraintestinal Dissemination Of Salmonella., Adarsh Gopinath

Electronic Theses and Dissertations

Salmonella appears in the bloodstream of mice in as little as 15 minutes after oral inoculation and establishes persistent colonies in the spleen and liver. While its pathway to blood is undetermined, this phenomenon is dependent on the activity of Salmonella pathogenicity island 2 (SPI-2) coded type III secretion system (T3SS) and CD18+ phagocytes. We hypothesize that dendritic cells associated with the basal face of the gut epithelium, that are naturally migratory and known to sample for luminal antigens directly transport Salmonella to the bloodstream. This process comprises of at least two phases, dissociation and reverse transmigration. We define dissociation …

An Analysis Of Crispr-Cas Gene Editing In Agriculture, Ashley Laliberte

Honors Scholar Theses

The CRISPR-Cas system is a promising form of gene editing, especially for the agriculture industry. The ability to make single-nucleotide edits within a gene of interest, without the need to introduce foreign DNA, is a powerful tool for designing healthier and more efficient crops and food animals. This system provides opportunity for increased nutritional value, decreased food waste, and more economically and environmentally sustainable food production. Though this biotechnology is facing mechanistic limitations due to off-target effects and inefficient homology-directed repair, vast improvements have already been made to improve its efficacy. The CRISPR-Cas system is already the most advanced form …

Cd11b+ Depletion Of Bone Marrow Derived Macrophages Negatively Affects Bone Fracture Healing, Sarah Hozain

Seton Hall University Dissertations and Theses (ETDs)

Hematoma formation and inflammation occurs during the beginning stages of fracture repair, which require the presence of innate cells such as macrophages. Macrophages are subdivided into proinflammatory, M1, or anti-inflammatory, M2, phenotypes with different functions, cytokine profiles, and surface markers. In this study, in vitro and in vivo models were used to deplete M1 macrophages, using Mac-1 Sap conjugated antibody (Mac1SAP), to determine the effects on fracture healing. In vitro study, primary macrophages isolated from mice femoral bone marrow were harvested and differentiated into M1 macrophages (+LPS), M2 macrophages (+IL-4), or undifferentiated then treated with either vehicle or 10pM Mac1SAP. …

Comparative Study Of The Effects Of Light On Photophore Ultrastructure From Two Families Of Deep-Sea Decapod Crustaceans: Oplophoridae And Sergestidae, Jamie E. Sickles

HCNSO Student Theses and Dissertations

Counterillumination, the mechanism by which pelagic species produce bioluminescence to replace the light blocked by their bodies to hide their silhouettes, has been known for over 100 years. However, little is known about how these animals are able to so precisely replicate the intensity of downwelling light. The recent discovery of opsins in photophores (Bracken-Grissom et al. 2020) suggests that these autogenic organs (i.e. non-bacterial) may be sensitive to light, in addition to their function of emitting visible light. The study presented here is 1) the first ultrastructural assessment of photophores in species Systellaspis debilis, Janicella spinicauda, Parasergestes armatus, …

486— Cell Cycle Effect On Hla Expression In Htb-4 And Mrc-5, Nick Turnquist, Andrew Kareeparampil

GREAT Day Posters

Human Leukocyte Antigen (HLA) is a gene that codes for cell-surface proteins that are the basis of our bodies’ immune response. We observed two cell lines, HTB-4 and a non-cancerous cell line known as MRC-5. Our purpose for this experiment was to observe the effect of the cell cycle on HLA expression for these cell lines. Two flasks were prepared for each cell line: one confluent and one non-confluent. Flow cytometry analysis was performed for each of the four flasks. The flow cytometry results of the MRC-5 cells indicate increased HLA expression in the confluent flask, in the G1 phase …