Cell and Developmental Biology Commons^™

Pathogenicity Of Acinetobacter Calcoaceticus, Kaitlan A. Sullivan

MUSC Theses and Dissertations

Acinetobacter is a genus of gram-negative bacteria that have been appearing frequently in hospitals contributing to infections in the blood, lungs, urinary tract, and other parts of the body. It infects patients with weakened immune systems that are placed on ventilators, after the use of catheters, or have any other open wounds produced by prolonged hospital stays. This genus of bacteria is problematic due to its high probability of becoming resistant to multiple classes of antibiotics. Thus, we are determining the pathogenicity of clinical isolates of Acinetobacter calcoaceticus using the organism Caenorhabditis elegans as a model.

We are testing …

The Role Of Cerium(Iii) In Bacterial Growth And The Microbial Transformation Of Aromatic Hydrocarbons, Shruti Sathish

Honors Theses

Biofilms are communities of surface-attached bacterial cells encased in an exopolymeric matrix. In this state, they are more resistant to antimicrobial treatment and can have adverse effects in medical, agricultural, and industrial settings. Whereas, as biocatalysts, biofilms from nonpathogenic bacteria enhance their performance and stability in catalysis. Unfortunately, there are several challenges when using bacteria in organic transformations due to their complex cellular chemistry. Trivalent lanthanide metals were discovered to serve regulatory roles in some bacterial catalytic processes, including those of Pseudomonas putida KT2440 (P. putida), a non-infectious Gram-negative bacterium. The main goal of our research is to use cerium(III) …

Principles Of Aaa+ Proteases, Samar Mahmoud

Doctoral Dissertations

ATPases associated with diverse cellular activities (AAA+) proteases in bacteria help maintain protein homeostasis by degrading misfolded and regulatory proteins. While a handful of protein targets for these proteases have been identified in Caulobacter crescentus and other organisms, more research is needed to elucidate mechanisms that govern substrate specificity. In the second chapter of this thesis, I will elaborate on how AAA+ substrate specificity is less rigid than previous work has suggested and how limiting ATP or mutations can alter substrate preferences of the ClpXP protease. In the third chapter, I will highlight our efforts to use a quantitative proteomics …

Resolving The Repression Pathway Of Virulence Gene Hila In Salmonella, Alexandra King, Lon Chubiz Phd, Brenda Pratte, Lauren Daugherty

Undergraduate Research Symposium

Salmonella is a relatively abundant, virulent species of bacteria that is most known for spreading gastrointestinal diseases through food. These illnesses result in approximately 1.35 million infections, including over 25,000 hospitalizations each year, in the U.S. alone (CDC.gov). As antibiotic resistance becomes an increasingly urgent public health problem, the importance of developing alternative treatment methods is only becoming more crucial. One of the genes responsible for this virulence is known as hilA. HilA is the main transcriptional regulator of Salmonella Pathogenicity Island-1 gene (UniProt). SPI-1 plays an important role in the invasion of Salmonella into epithelial cells. The proteins encoded …

Structural Formation And Functionality Of The Spoiie Protein In Clostridium Difficile, Rachel Siebenmorgen

Biological Sciences Undergraduate Honors Theses

My research efforts were directed toward the structure and functionality of a protein, designated SpoIIE [11], which previously has been shown to regulate C. difficile peptide utilization in addition to sporulation and toxin release. The SpoIIE protein is found in all spore-forming gram-positive bacteria and is a crucial component in activation of transcription factors and septum formation thus affecting the regulation and timing of spore development [11,12]. A truncated form of the SpoIIE protein named SpoIIE∆TM was used in an effort to determine the structure of an important but uncharacterized linker domain of the protein. If the structure and specific …

Evaluation Of Selected Nuproxa Feed Additives For Protection Against Lameness And Improving The Wellbeing Of Broilers In A Lameness Challenge Model, Jaren Hernandez

Biological Sciences Undergraduate Honors Theses

A common concern in commercial industries profiting from broiler chickens is their inability to yield a completely healthy flock. Bacterial Chondronecrosis with Osteomyelitis poses a threat to these companies by impairing the birds' proximal and distal tibial and femoral heads. In 2000, roughly 17.3% of broiler chickens were affected by Bacterial Chondronecrosis with Osteomyelitis which caused a substantial economic loss in the industry (McNammee et al., 2000). BCO cannot be tied to a particular bacterial species; this experiment focuses on evaluating the efficacy of certain commercial feed additives while determining the bacterial species causing BCO in this set of the …

Assessment Of Diversity Of Antimicrobial Resistance Phenotypes And Genotypes Of Mannheimia Haemolytica Isolates From Bovine Nasopharyngeal Swabs, Hannah F. Carter, Robert W. Wills, Matthew A. Scott, Alexis C. Thompson, Randall S. Singer, John Dustin Loy, Brandi B. Karisch, William B. Epperson, Amelia R. Woolums

School of Veterinary and Biomedical Sciences: Faculty Publications

The threat of bovine respiratory disease (BRD) for cattle operations is exacerbated by increasing prevalence of antimicrobial resistance (AMR) in Mannheimia haemolytica, a leading cause of BRD. Characterization of AMR in M. haemolytica by culture and susceptibility testing is complicated by uncertainty regarding the number of colonies that must be selected to accurately characterize AMR phenotypes (antibiograms) and genotypes in a culture. The study objective was to assess phenotypic and genotypic diversity of M. haemolytica isolates on nasopharyngeal swabs (NPS) from 28 cattle at risk for BRD or with BRD. NPS were swabbed onto five consecutive blood agar plates; …

Tick Extracellular Vesicles Enable Arthropod Feeding And Promote Distinct Outcomes Of Bacterial Infection, Adela S. Oliva Chávez, Xiaowei Wang, Liron Marnin, Nathan K. Archer, Holly L. Hammond, Erin E. Mcclure Carroll, Dana K. Shaw, Brenden G. Tully, Amanda D. Buskirk, Shelby L. Ford, L. Rainer Butler, Preeti Shahi, Kateryna Morozova, Cristina C. Clement, Lauren Lawres, Anya J. O'Neal, Choukri Ben Mamoun, Kathleen L. Mason, Brandi E. Hobbs, Glen A. Scoles, Eileen M. Barry, Daniel E. Sonenshine, Utpal Pal, Jesus G. Valenzuela, Marcelo B. Sztein, Marcela F. Pasetti, Michael L. Levin, Michail Kotsyfakis, Steven M. Jay, Jason F. Huntley, Lloyd S. Miller, Laura Santambrogio, Joao H.F. Pedra

Biological Sciences Faculty Publications

Extracellular vesicles are thought to facilitate pathogen transmission from arthropods to humans and other animals. Here, we reveal that pathogen spreading from arthropods to the mammalian host is multifaceted. Extracellular vesicles from Ixodes scapularis enable tick feeding and promote infection of the mildly virulent rickettsial agent Anaplasma phagocytophilum through the SNARE proteins Vamp33 and Synaptobrevin 2 and dendritic epidermal T cells. However, extracellular vesicles from the tick Dermacentor andersoni mitigate microbial spreading caused by the lethal pathogen Francisella tularensis. Collectively, we establish that tick extracellular vesicles foster distinct outcomes of bacterial infection and assist in vector feeding by acting …

The Resistive Barrier Discharge: A Brief Review Of The Device And Its Biomedical Applications, Mounir Laroussi

Electrical & Computer Engineering Faculty Publications

This paper reviews the principles behind the design and operation of the resistive barrier discharge, a low temperature plasma source that operates at atmospheric pressure. One of the advantages of this plasma source is that it can be operated using either DC or AC high voltages. Plasma generated by the resistive barrier discharge has been used to efficiently inactivate pathogenic microorganisms and to destroy cancer cells. These biomedical applications of low temperature plasma are of great interest because in recent times bacteria developed increased resistance to antibiotics and because present cancer therapies often are accompanied by serious side effects. Low …

Phenotypic Switching Of Bacterial Cells In Extreme Environments, Sudip Nepal

Graduate Theses and Dissertations

A large number of terrestrial microbial lives thrive in extremes of environmental conditions, including extremes of pressure, temperature, salinity, pH, and a combination of them. For example, all the marine biomass thrive at high hydrostatic pressure depending on depth. The temperature in the ocean can be very high near the hydrothermal vents and salinity and pH depends on the composition of salt in the surrounding areas. On the surface, hot springs, lakes and geysers provide high temperature conditions, while many places are permafrost regions with subzero temperatures. There is an emerging body of work on the viability, genomics, and metagenomics …

Triggers Of Inflammatory Heart Disease, Ninaad Lasrado, Bharathi Yalaka, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Inflammatory heart disease (IHD) is a group of diseases that includes pericarditis, myocarditis, and endocarditis. Although males appear to be more commonly affected than females, IHD can be seen in any age group. While the disease can be self-limiting leading to full recovery, affected individuals can develop chronic disease, suggesting that identification of primary triggers is critical for successful therapies. Adding to this complexity, however, is the fact that IHD can be triggered by a variety of infectious and non-infectious causes that can also occur as secondary events to primary insults. In this review, we discuss the immunological insights into …

Influence Of Developmental Stage, Habitat, And Captivity On Thecutaneous Bacterial Communities Of Eastern Hellbenders (Cryptobranchus Alleganiensis Alleganiensis) In West Virginia, Rachel Fern Arrick

Theses, Dissertations and Capstones

Microbes inhabit virtually all surfaces of multicellular animal hosts, with microbial cells outnumbering the hosts’ own cells 10:1. Symbiont microbes, collectively referred to as the microbiome, can have profound impacts on the metabolism, development, behavior, and disease resistance of their multicellular hosts. Because the community structure of symbiont bacteria can influence host health, the characterization of amphibian microbiomes is becoming an increasingly important tool for future conservation in the face of global amphibian declines. Eastern hellbenders are good candidates for a microbiome study because they have seen substantial declines in recent decades and learning more about the environmental and physiological …

Dual-Functional-Tag-Facilitated Protein Labeling And Immobilization, Xinyi Zhang, Wei Lu, Kevin Kwan, Dibakar Bhattacharyya, Yinan Wei

Chemistry Faculty Publications

An important strategy in the construction of biomimetic membranes and devices is to use natural proteins as the functional components for incorporation in a polymeric or nanocomposite matrix. Toward this goal, an important step is to immobilize proteins with high efficiency and precision without disrupting the protein function. Here, we developed a dual-functional tag containing histidine and the non-natural amino acid azidohomoalanine (AHA). AHA is metabolically incorporated into the protein, taking advantage of the Met-tRNA and Met-tRNA synthetase. Histidine in the tag can facilitate metal-affinity purification, whereas AHA can react with an alkyne-functionalized probe or surface via well-established click chemistry. …

Optimization Of A Genomic Editing System Using Crispr/Cas9-Induced Site-Specific Gene Integration, Jillian L. Mccool Ms., Nick Hum, Gabriela G. Loots

STAR Program Research Presentations

The CRISPR-Cas system is an adaptive immune system found in bacteria which helps protect against the invasion of other microorganisms. This system induces double stranded breaks at precise genomic loci (1) in which repairs are initiated and insertions of a target are completed in the process. This mechanism can be used in eukaryotic cells in combination with sgRNAs (1) as a tool for genome editing. By using this CRISPR-Cas system, in addition to the “safe harbor locus,” ROSAβ26, the incorporation of a target gene into a site that is not susceptible to gene silencing effects can be achieved through few …

Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli

Doctoral Dissertations

Finding and treating cancer is difficult due to limited sensitivity and specificity of current detection and treatment strategies. Many chemotherapeutic drugs are small molecules that are limited by diffusion, making it difficult to reach cancer sites requiring high doses that lead to systemic toxicity and off-target effects. Tomographic detection techniques, like PET, MRI and CT, are good at identifying macroscopic lesions in the body but are limited in their ability to detect microscopic lesions. Biomarker detection strategies are extremely sensitive and able to identify ng/ml concentrations of protein, but are poor at discriminating between healthy and disease state levels due …

Examining Phage Infection Utilizing Mycobacterium Smegmatis, Tanya L. Riddick

Undergraduate Research

Bacteriophages, also known as phages, are viruses that are ubiquitous and survive and replicate within the host of the bacterial cell, Mycobacterium smegmatis. They are considered one of the most abundant organisms on earth (10³¹). Structurally, they are 100-200nm in size and consist of a protein encapsulated head that contains DNA or RNA, a tail sheath and tail fibers. This research consisted of examining phage infection, by re-isolating a novel phage, Tango. Tango was originally isolated genetically in 2013 by a previous ISBT student, Anna Maccarrone. The phage was sent to genetic sequencing but two phages were discovered, …

Characterization Of The Interaction Between Ftsz-Ring Stabilizer Zapc And The Conserved Protease Clpxp In E. Coli Cytokinesis, Andrea Cardenas Arevalo

Dissertations and Theses

Cell division in bacteria requires the assembly of a macromolecular protein machinery at midcell that is spatiotemporally regulated during the bacterial cell cycle. Central to the process of division is the assembly of a cytokinetic ring-like structure, termed the Z-ring, formed by polymers of the essential tubulin-homolog FtsZ. Polymerization of FtsZ is regulated by a number of proteins that bind FtsZ and regulate its assembly/disassembly mechanisms. Despite being extensively studied, the molecular nature of the diverse protein-protein interactions that regulate the assembly of FtsZ during early stages of division is not well understood. Here, I focus on an FtsZ-ring stabilizer …

Structure-Function Analysis Of Zapc, An Ftsz-Ring Stabilizer, In Escherichia Coli Cytokinesis, Lukasz Tchorzewski

Dissertations and Theses

In Escherichia coli, cell division is defined by the polymerization and constriction of a cytokinetic ring (Z ring) formed by FtsZ, a tubulin-like GTPase, at midcell. Division also involves the formation of a multi-protein complex at midcell known as the divisome. Several divisome proteins promote the assembly/disassembly processes of FtsZ, thereby exercising spatiotemporal control over division. Among FtsZ regulatory proteins are the FtsZ ringassociated proteins (Zap), which either directly or indirectly stabilize the Z-ring by increasing lateral interactions amongst FtsZ protofilaments in the Z-ring. ZapA-D are recruited during early cytokinesis and have overlapping functions in stabilizing FtsZ at midcell, but …

Divergent Mechanisms Of Interaction Of Helicobacter Pylori And Campylobacter Jejuni With Mucus And Mucins, Julie Ann Naughton, Karina Mariño, Brendan Dolan, Colm Reid, Ronan Gough, Mary Gallagher, Michelle Kilcoyne, Jared Gerlachscience,, Lokesh Joshi, Pauline Rudd, Stephen Carrington, Billy Bourke, Marguerite Clyne

Articles

Helicobacter pylori and Campylobacter jejuni colonize the stomach and intestinal mucus, respectively. Using a combination of mucus-secreting cells, purified mucins, and a novel mucin microarray platform, we examined the interactions of these two organisms with mucus and mucins. H. pylori and C. jejuni bound to distinctly different mucins. C. jejuni displayed a striking tropism for chicken gastrointestinal mucins compared to mucins from other animals and preferentially bound mucins from specific avian intestinal sites (in order of descending preference: the large intestine, proximal small intestine, and cecum). H. pylori bound to a number of animal mucins, including porcine stomach mucin, but …

Use Of Image Cytometry For Quantification Of Pathogenic Fungi In Association With Host Cells, Charlotte A. Berkes, Leo Li-Ying Chan, Alisha Wilkinson, Benjamin Paradis

Biology Faculty Publications

Studies of the cellular pathogenesis mechanisms of pathogenic yeasts such as Candida albicans, Histoplasma capsulatum, and Cryptococcus neoformans commonly employ infection of mammalian hosts or host cells (i.e. macrophages) followed by yeast quantification using colony forming unit analysis or flow cytometry. While colony forming unit enumeration has been the most commonly used method in the field, this technique has disadvantages and limitations, including slow growth of some fungal species on solid media and low and/or variable plating efficiencies, which is of particular concern when comparing growth of wild-type and mutant strains. Flow cytometry can provide rapid quantitative information regarding yeast …

Host Pathogen Interactions: Is Arabidopsis Thaliana Remembered By Its Nemesis Pseudomonas Syringae?, Daniel Z. Kreiser

Lawrence University Honors Projects

Plants contain innate immune systems that deter pathogen infection. Pattern recognition receptors bind microbe-associated molecular patterns (MAMPs), triggering immunity. MAMPs are proteins exclusive to pathogens that are typically indispensable for their survival. For this reason, MAMPs cannot be mutated or removed without causing pathogen death. However, this does not necessitate constitutive expression of MAMPs. In this study, the MAMP response of Arabidopsis thaliana was utilized to determine differential detection of MAMPs expressed by Pseudomonas syringe pv. tomato DC3000 when pretreated with A. thaliana. Results demonstrated that more MAMPs are detected when P. syringae had previously encountered A. thaliana, …

Novel Phenol Soluble Modulin Derivatives In Community-Associated Methicillin- Resistant Staphylococcus Aureus Identified Through Imaging Mass Spectrometry., D. Gonzalez, Cheryl Okumura, A. Hollands, R. Kersten, K. Akong-Moore, M. Pence, C. Malone, J. Derieux, B. Moore, A. Horswill, J. Dixon, P. Dorrestein, V. Nizet

Cheryl Okumura

Staphylococcus aureus causes a wide range of human disease ranging from localized skin and soft tissue infections to potentially lethal systemic infections. S. aureus has the biosynthetic ability to generate numerous virulence factors that assist in circumventing the innate immune system during disease pathogenesis. Recent studies have uncovered a set of extracellular peptides produced by community-associated methicillin-resistant S. aureus (CA-MRSA) with homology to the phenol-soluble modulins (PSMs) from Staphylococcus epidermidis. CA-MRSA PSMs contribute to skin infection and recruit and lyse neutrophils, and truncated versions of these peptides possess antimicrobial activity. In this study, novel CA-MRSA PSM derivatives were discovered by …

Kinetics And Relative Importance Of Phosphorolytic And Hydrolytic Cleavage Of Cellodextrins And Cellobiose In Cell Extracts Of Clostridium Thermocellum, Yie.-Heng P. Zhang, Lee R. Lynd

Dartmouth Scholarship

Rates of phosphorolytic cleavage of 􏰁-glucan substrates were determined for cell extracts from Clostridium thermocellum ATCC 27405 and were compared to rates of hydrolytic cleavage. Reactions with cellopentaose and cellobiose were evaluated for both cellulose (Avicel)- and cellobiose-grown cultures, with more limited data also obtained for cellotetraose. To measure the reaction rate in the chain-shortening direction at elevated temper- atures, an assay protocol was developed featuring discrete sampling at 60°C followed by subsequent analysis of reaction products (glucose and glucose-1-phosphate) at 35°C. Calculated rates of phosphorolytic cleavage for cell extract from Avicel-grown cells exceeded rates of hydrolytic cleavage by >20-fold …

Development And Lipid Composition Of The Harpacticoid Copepod Nitocra Spinipes Reared On Different Diets, Gregory M. Weiss, George B. Mcmanus, H. Rodger Harvey

OES Faculty Publications

We reared the harpacticoid copepod Nitocra spinipes on diets of bacteria, a diatom, or a macroalga, evaluating survivorship and growth in short-term (≤ 1 generation) experiments. Lipid content of the copepods and their diets was measured and used as an index of nutrition. Although growth, survivorship and lipid content of N. spinipes were significantly greater when fed the diatom, which had the highest lipid content of the 3 diets, the copepod was able to develop from egg to adult when fed a lipid- poor bacterial diet. Furthermore, this species was able to go through developmental molts without the addition of …

Trophic Transfer Of Fatty Acids, Sterols, And A Triterpenoid Alcohol Between Bacteria, A Ciliate, And The Copepod Acartia Tonsa, Melissa C. Ederington, George B. Mcmanus, H. Rodger Harvey

OES Faculty Publications

The incorporation of lipids into the copepod Acartia tonsa and its eggs was measured when it was fed either a bacterivorous ciliate (Pleuronema sp.) or a diatom (Thalassiosira weissflogii). Egg production was 10-fold higher on the diatom diet, whereas hatch success of eggs was the same for algal and ciliate diets. Adult copepods fed diatoms contained more total fatty acid and sterols than copepods fed the ciliate diet, and individual lipids reflected the dietary source. Eggs from diatom-fed copepods had fewer fatty acids but more sterols than eggs from copepods on a ciliate diet. Ciliate-fed copepods and …

An Electrochemical Method Of Measuring The Oxidation Rate Of Ferrous To Ferric Iron With Oxygen In The Presence Of Thiobacillus Ferrooxidans, David J. Oliver, B. Pesic, P. Wichlacz

David J. Oliver

The oxidation of Fe2+ with oxygen in sulfate solutions was studied in the presence of T. ferrooxidans. To measure the chemical activity of bacteria, and the oxidation rate of iron, the redox potentials of solutions were continuously monitored during the experiments. The redox potentials were simultaneously monitored on the platinum and pyrite indicator electrodes. The redox potential versus time curves were further used to calculate the basic kinetic parameters, such as the reaction orders, the activation energy, and the frequency factor. It was found that under atmospheric conditions, and at Fe2+ < 0.001M, T < 25°C, and at pH above 2.2, the oxidation of iron is governed by the following rate expression: [equation image] Below pH = 2.2, the oxidation rate is independent of H+ Concentration.