Life Sciences Commons^™

Expression Of Acetate Permease-Like (Apl) Genes In Subsurface Communities Of Geobacter Species Under Fluctuating Acetate Concentrations, Derek Lovley, Hila Elifantz, Lucie N'Guessan, Paula Mouser, Kenneth Williams, Michael Wilkins, Carla Risso, Dawn Holmes, Philip Long

Carla Risso

The addition of acetate to uranium-contaminated aquifers in order to stimulate the growth and activity of Geobacter species that reduce uranium is a promising in situ bioremediation option. Optimizing this bioremediation strategy requires that sufficient acetate be added to promote Geobacter species growth. We hypothesized that under acetate-limiting conditions, subsurface Geobacter species would increase the expression of either putative acetate symporters genes (aplI and aplII). Acetate was added to a uranium-contaminated aquifer (Rifle, CO) in two continuous amendments separated by 5 days of groundwater flush to create changing acetate concentrations. While the expression of aplI in monitoring well D04 (high …

Optimization Of Application Of Delactosed Whey Permeate Treatment To Extend The Shelf-Life Of Fresh Cut Tomato Using Response Surface Methodology., Lubna Ahmed, Ana Belen Martin-Diana, Daniel Rico, Catherine Barry-Ryan

Articles

Optimization of delactosed whey permeate (DWP) treatment for fresh-cut tomato was accomplished by evaluating different quality, nutritional and microbial markers. Response surface methodology was applied to obtain polynomial model equations. DWP concentration (0 - 5 %) and storage (0 - 10 days) were used as independent factors in order to optimize the process. The analyses showed that increases in DWP concentration extended the quality of the fresh-cut tomato significantly (p3 % were scored unacceptable by the sensory panel due to perceived off-odours. DWP treatment also improved retention of ascorbic acid and lycopene over storage. The total aerobic counts and yeast …

Application Of Cyclic Voltammetry To Investigate Enhanced Catalytic Current Generation By Biofilm-Modified Anodes Of Geobacter Sulfurreducens Strain Dl1 Vs. Variant Strain Kn400, Sarah M. Strycharz, Anthony P. Malanoski, Rachel M. Snider, Hana Yi, Derek Lovley, Leonard M. Tender

Derek Lovley

A biofilm of Geobacter sulfurreducens will grow on an anode surface and catalyze the generation of an electrical current by oxidizing acetate and utilizing the anode as its metabolic terminal electron acceptor. Here we report qualitative analysis of cyclic voltammetry of anodes modified with biofilms of G. sulfurreducens strains DL1 and KN400 to predict possible rate-limiting steps in current generation. Strain KN400 generates approximately 2 to 8-fold greater current than strain DL1 depending upon the electrode material, enabling comparative electrochemical analysis to study the mechanism of current generation. This analysis is based on our recently reported electrochemical model for biofilm-catalyzed …

Discovery And Characterization Of An Antibiotic From The Soil Bacterium Bacillus Sp., Thomas S. Barber

Electronic Theses and Dissertations

Many important antibiotics have become nearly obsolete due to the rise of antibiotic resistant pathogens. Rhodococcus, an actinomycete related to the prolific antibiotic producing genus Streptomyces, harbors over 30 genes for secondary metabolism that could be involved in antibiotic production. Several antibiotics have already been reported for Rhodococcus, suggesting the genus may be a good source for new inhibitory compounds. Fifty four soil bacteria were isolated using enrichment culture techniques (including 37 Rhodococcus) and screened for antibiotic producers. BTHX2, a species of Bacillus was found to have activity against Micrococcus luteus and Rhodococcus erythropolis. BTHX2 …

Modulation Of Alpha-Subunit Visit-Dg Sequence Residues Ser-347, Gly-351 And Thr-349 In The Catalytic Sites Of Escherichia Coli Atp Synthase., Laura Elaine Brudecki

Electronic Theses and Dissertations

Binding of inorganic phosphate (P_i) in ATP synthase catalytic sites is a crucial step for the synthesis of adenosine-5'-triphosphate (ATP). ATP is the fundamental means of cellular energy in almost every organism, and in order to gain insight into the regulation of ATP catalysis, critical amino acid residues responsible for binding P_i must be identified. Here, we investigate the role of highly conserved α-subunit VISIT-DG sequence residues αSer-347, αGly-351, and αThr-349 in P_i binding. Mutations αS347A/Q, αG351Q, αT349A/D/R, βR182A, and αT349R/βR182A were generated via site directed mutagenesis. Results from biochemical assays showed that αSer-347 is required …

H-Ns Binding And Repression Of The Ctx Promoter In Vibrio Cholerae, Emily A. Stonehouse, Robin R. Hulbert, Melinda B. Nye, Karen Skorupski, Ronald K. Taylor

Dartmouth Scholarship

Expression of the ctx and tcp genes, which encode cholera toxin and the toxin coregulated pilus, the Vibrio cholerae O1 virulence determinants having the largest contribution to cholera disease, is repressed by the nucleoid-associated protein H-NS and activated by the AraC-like transcriptional regulator ToxT. To elucidate the molecular mechanism by which H-NS controls transcription of the ctxAB operon, H-NS repression and binding were characterized by using a promoter truncation series, gel mobility shift assays, and DNase I footprinting. Promoter regions found to be important for H-NS repression correlated with in vitro binding. Four main H-NS binding regions are present at …

Campylobacter Ureolyticus: An Emerging Gastrointestinal Pathogen?, Susan Bullman, Daniel Corcoran, James O'Leary, Brigid Lucey, Deirdre Byrne, Roy D. Sleator

Department of Biological Sciences Publications

A total of 7194 faecal samples collected over a 1-year period from patients presenting with diarrhoea were screened for Campylobacter spp. using EntericBios, a multiplex-PCR system. Of 349 Campylobacter-positive samples, 23.8% were shown to be Campylobacter ureolyticus, using a combination of 16S rRNA gene analysis and highly specific primers targeting the HSP60 gene of this organism. This is, to the best of our knowledge, the first report of C. ureolyticus in the faeces of patients presenting with gastroenteritis and may suggest a role for this organism as an emerging enteric pathogen.

Analysis Of Polymorphism In Human Cytomegalovirus (Hcmv) Chemokine, Vcxcl-1, And Its Role In Cellular Activation, Jinho Heo

Doctoral Dissertations

The human cytomegalovirus (HCMV) viral chemokine gene, UL146, shows a high degree of variability in clinical isolates. The UL146-produced viral chemokine, vCXCL-1, has homology to CXC chemokines and is predicted to be an immune modulator that may contribute to the pathogenesis of HCMV infections. In the analysis of clinical isolates from congenitally infected infants, we found 11 distinct vCXCL-1 clades. Although the four cysteine residues that create two disulfide bonds providing the essential structure for CXC chemokines,are conserved, the N-loop region, which is important for receptor binding and activation, was hypervariable. One clade also contained a modified glutamic acid-leucine-arginine (ELR) …

Characterization Of The Extracellular Proteome Of A Natural Microbial Community With An Integrated Mass Spectrometric / Bioinformatic Approach, Brian Keith Erickson

Doctoral Dissertations

Proteomics comprises the identification and characterization of the complete suite of expressed proteins in a given cell, organism or community. The coupling of high performance liquid chromatography (LC) with high throughput mass spectrometry (MS) has provided the foundation for current proteomic progression. The transition from proteomic analysis of a single cultivated microbe to that of natural microbial assemblages has required significant advancement in technology and has provided greater biological understanding of microbial community diversity and function.

To enhance the capabilities of a mass spectrometric based proteomic analysis, an integrated approach combining bioinformatics with analytical preparations and experimental data collection was …

Ultraviolet Light As An Adjunct Treatment To Pasteurization For Microbial Reduction In Milk Intended For Extended Shelf Life, Jacob Zonneveld

Dairy Science

The objective of this study was to determine whether ultraviolet light treatment of milk could be used in adjunct with pasteurization and to determine whether the microbial count in bulk tank raw milk was sufficient enough to receive significant results. Samples were collected from a bulk tank filled with 1600 gallons of raw Holstein and Jersey milk from the Cal Poly Dairy throughout the course of a week. Control (library) samples were also taken from the raw milk tank and stored without air or agitation for comparison. These samples were plated each day and examined 24 and 48 hours after …

Bioinformatic Analysis Of Gene Regulation In The Metal-Reducing Bacteria Family, Derek Lovley, Julia Krushkal, Ronald M. Adkins, Yanhua Qu, Jeanette Peeples, Sreedhar Sontineni, Ching Leang, Peter Brown, Nelson D. Young, Toshiyuki Ueki, Katy Juarez

Derek Lovley

Knowledge of how structural genome differences among microorganisms lead to variation in gene regulation is fundamentally important for our understanding of the functioning of gene regulatory pathways and their individual components. This knowledge is also necessary for our better understanding of the genomic changes leading to adaptation to diverse and changing environments. Our research focuses on Geobacteraceae, a metal-reducing family of delta-Proteobacteria, which are capable of harvesting electricity from organic matter and environmental bioremediation of organic and metal pollutants. We are investigating molecular mechanisms which allow these species to adapt and regulate their responses to environmental stimuli, which result in …

Transcriptional Regulation Of Shigella Virulence Plasmid-Encoded Genes By Virb And Crp, Christopher Thomas Hensley

UNLV Theses, Dissertations, Professional Papers, and Capstones

Shigella flexneri is a species of Gram-negative intracellular pathogens that causes bacillary dysentery in humans. Shigella relies on the precise transcriptional regulation of virulence genes, encoded by a large virulence plasmid, for invasion and infection of human colonic epithelial cells. The transcription of most identified virulence genes are regulated through a cascade controlled by the primary regulator of virulence genes, VirF, and the global transcriptional regulator, VirB. Currently, few studies have addressed how individual Shigella virulence genes are precisely regulated for optimal expression during specific stages of pathogenesis and within the constraints of the regulatory cascade. This work addresses how …

Agrobacterium Virb10 Contributions To Type Iv Substrate Secretion, T-Pilus Biogenesis, And Outer Membrane Pore Formation, Isaac Garza

Dissertations & Theses (Open Access)

The VirB/D4 type IV secretion system (T4SS) of Agrobacterium tumefaciens functions to transfer substrates to infected plant cells through assembly of a translocation channel and a surface structure termed a T-pilus. This thesis is focused on identifying contributions of VirB10 to substrate transfer and T-pilus formation through a mutational analysis. VirB10 is a bitopic protein with several domains, including a: (i) cytoplasmic N-terminus, (ii) single transmembrane (TM) α-helix, (iii) proline-rich region (PRR), and (iv) large C-terminal modified β-barrel. I introduced cysteine insertion and substitution mutations throughout the length of VirB10 in order to: (i) test a predicted transmembrane topology, (ii) …

The Immune Inhibitor A1 Protease Of Bacillus Anthracis, Kathryn Pflughoeft

Dissertations & Theses (Open Access)

Bacillus anthracis, an organism ubiquitous in the soil and the causative agent of anthrax, utilizes multiple mechanisms to regulate secreted factors; one example is the activity of secreted proteases. One of the most abundant proteins in the culture supernates of B. anthracis is the Immune Inhibitor A1 (InhA1) protease. Here, I demonstrate that InhA1 modulates the abundance of approximately half of the proteins secreted into the culture supernates, including substrates that are known to contribute to the ability of the organism to cause virulence. For example, InhA1 cleaves the anthrax toxin proteins, PA, LF, and EF. InhA1 also targets …

Sequencing And Functional Analysis Of A Multi-Component Dioxygenase From Pah-Degrading Sphingomonas Paucimobilis Epa505, Renuka Miller

All Dissertations

Polycyclic aromatic hydrocarbons (PAHs) are hydrophobic organic compounds consisting of two or more fused benzene rings. PAHs derive from many different sources including petroleum refining, wood treatment, and coal coking industries. Because of their structural stability and water insolubility, PAHs are extremely resistant to degradation. These compounds are also believed to have mutagenic, carcinogenic, and teratogenic effects. Therefore, there are currently 16 PAH compounds on the EPA's list of priority pollutants.
Many species of bacteria have the ability to breakdown these persistent pollutants. However, bioremediation strategies using these organisms have many unresolved issues. While laboratory experiments can easily demonstrate the …

The Molecular Mechanism Of Viral Transactivator Rta-Mediated Kaposi’S Sarcoma-Associated Herpesvirus (Kshv) Lytic Replication, Hui-Ju Wen

School of Biological Sciences: Dissertations, Theses, and Student Research

Kaposi’s sarcoma-associated herpesvirus (KSHV) is a member of the gamma-herpesvirinae subfamily and displays two distinct life phases, latency and lytic replication. Infection with KSHV mostly results in a latent state. A small population of infected cells can spontaneously undergo lytic phase, which is marked by abundant viral gene expression and production of infectious viral progeny. Lytic replication is important for transmission of KSHV in the population and development of diseases. RTA (replication and transcription activator) is a master regulator of KSHV lytic replication. Expression of RTA alone is sufficient to disrupt KSHV latency and initiate the lytic replication cascade. The …

Biofuel Ethanol Production By Saccharomyces Bayanus, The Champagne Yeast, Kristen Miller

All Theses

The importance of biofuel ethanol is growing as the demand for clean, renewable fuels produced from non-food sources increases. The United States relies mainly on corn and the yeast Saccharomyces cerevisiae for the production of ethanol. A shift to cellulosic feedstocks, as the main source of biomass for ethanol production, would alleviate the pressure on farmers to produce corn for both the food industry and the ethanol industry. Example cellulosic feedstocks include switchgrass, sorghum, and canary grass. The cellulosic feedstocks are typically grown on land that cannot support economic food production, and thus lay unused.
For cellulosic feedstocks to be …

Bovine Colostrum, Mary Hayes

All Theses

Colostrum is the first milk produced by mammals within a 24 to 72 hour period after parturition. Bovine colostrum is sold commercially as a nutraceutical product and its manufacturers purport numerous health benefits of the product for treating gastrointestinal disorders, respiratory tract disorders, viral and bacterial infections, and promoting tissue repair.
Four commercially available colostrum products, two whey, and one non-fat dry milk were obtained on separate days. These products were analyzed for fat, protein, ash, moisture and dry matter content. One colostrum product did not adhere to label claims in that it contained a mean of 19.1% fat whereas …

In Vitro Boosting And Expansion Of Tumor Infiltrating Killer T Cells, Chunlei Mei

All Theses

Immune cell tumor infiltration represents one of the mechanisms that the immune system responds to tumor cells. The tumor infiltrated immune cells include lymphocytes (TIL), dendritic cells (DCs), macrophages, and natural killer (NK) cells. Although TILs have been extensively studied in order to develop adoptive transfer based immunotherapies, how to effectively isolate, culture, and in vitro boosting the killer cells from TILs remains a challenge. Meanwhile, OK-432, a heat and penicillin-treated lyophilized preparation of the low-virulence strain of Streptococcus pyogenes, has been shown to exhibit immunomodulatory activities and potential antitumor therapeutic function, including the activation of DCs, neutrophils, macrophages, lymphocytes, …

Staphylococcus Aureus Virulence Factors Synthesis Is Controlled By Central Metabolism, Yefei Zhu

School of Veterinary and Biomedical Sciences: Dissertations, Theses, and Student Research

Staphylococcus aureus is a versatile pathogen that can survive in diverse host environments. This versatility depends on its ability to sense nutrients and respond by modulating gene expression, including the synthesis of virulence determinants. In addition to its ability to synthesize virulence factors, the capacity of S. aureus to form biofilms is an important mediator of virulence in certain infections. Biofilms are a complex aggregation of bacteria commonly encapsulated by an adhesive exopolysaccharide matrix (polysaccharide intercellular adhesin; PIA). To study S. aureus biofilm formation, we assessed the metabolic requirements of S. aureus growing in a biofilm and found the bacteria …

Stress-Induced Targeting Of Molecular Chaperones In The Yeast Saccharomyces Cerevisiae, Hugo Tapia

Dissertations & Theses (Open Access)

The eukaryotic stress response is an essential mechanism that helps protect cells from a variety of environmental stresses. Cell death can result if cells are not able to properly adapt and protect themselves against adverse stress conditions. Failure to properly deal with stress has implications in human diseases including neurodegenerative disorders and distinct cancers, emphasizing the importance of understanding the eukaryotic stress response in detail. As part of this response, expression of a battery of heat shock proteins (HSP) is induced, which act as molecular chaperones to assist in the repair or triage of unfolded proteins. The 90-kDa HSP (Hsp90) …

Characterization Of The Agrobacterium Tumefaciens Virb2 Pilin Of The Virb/D4 Type Iv Secretion System, Jennifer Kerr

Dissertations & Theses (Open Access)

The Agrobacterium tumefaciens VirB/D4 type IV secretion system (T4SS) delivers oncogenic T-DNA and effector proteins to susceptible plant cells. This leads to the formation of tumors termed Crown Galls. The VirB/D4 T4SS is comprised of 12 subunits (VirB1 to VirB11 and VirD4), which assemble to form two structures, a secretion channel spanning the cell envelope and a T-pilus extending from the cell surface. In A. tumefaciens, the VirB2 pilin subunit is required for assembly of the secretion channel and is the main subunit of the T-pilus. The focus of this thesis is to define key reactions associated with the T4SS …

Development And Characterization Of Novel Sub-Coatings With Polymorphic Brookite Titania Nanoparticles: Enhanced Uv/Vis Photocatalytic Antibacterial And Anticancer Properties, Donna Weinbrenner

All Dissertations

ABSTRACT
Nosocomial and community infections and biofilm formation from bacteria has increased significantly through adaptation combined with overuse of broad spectrum antibiotics. Because the world population continues to escalate, hospitals and long-term managed care will also escalate, thereby increasing transmission of infections, lowering patients' quality of health. Another disease on the rise throughout the world is skin cancer. A treatment modality that would cause less deleterious effects on the patient would be ideal. These two seemingly different issues could be solved with one product. First an inexpensive, safe, and non-selective antimicrobial surface coating would enhance the hospital arena and a …

Efficient Separation And Sensitive Detection Of Listeria Monocytogenes Using Magnetic Nanoparticles, Microfluidics And Interdigitated Microelectrode Based Impedance Immunosensor, Damira Ashimkhanovna Kanayeva

Graduate Theses and Dissertations

Listeria monocytogenes continues to be a major foodborne pathogen that causes food poisoning and sometimes death in immunosuppressed people and abortion in pregnant women. Nanoparticles have recently drawn attentions for use in immunomagnetic separation techniques due to their greater surface area/volume ratio and better stability against sedimentation in the absence of a magnetic field. Interdigitated microelectrodes and microfluidics make material transfer more efficient and biological/chemical interaction between the surface and solution phase much quicker. Magnetic nanoparticles (Fe3O4) with a 30 nm diameter were functionalized with rabbit anti-L. monocytogenes antibodies via biotin-streptavidin bonds and then amalgamated with target bacterial cells …

Analysis Of The Clear Plaque Phenotype Of The Bacteriophage Hk75, Phani Chandrika Kunapuli

Masters Theses & Specialist Projects

The growth of bacteriophage HK75 is inhibited by specific mutations in the zinc binding domain of the host RNA polymerase beta prime subunit. It shares this rare property with bacteriophage HK022 and other phages that use RNA mediated antitermination to promote early gene expression. Recent genomic analysis of HK75 and HK022 has confirmed the relatedness of these two phages and place HK75 in the lambdoid family of bacteriophages. Lambdoid phages are temperate and can adopt a lytic or lysogenic lifestyle upon infection of a suitable host. However, HK75 only forms clear plaques and thus appears to be defective in its …

Study Of Population Diversity Of Toxoplasma Gondii, Debashree Majumdar

Masters Theses

Toxoplasma gondii, the causal agent of toxoplasmosis, is an important water and food borne protozoan parasite. T. gondii was previously shown to have a distinct clonal population structure composed of Type I, II and III lineages in North America and Europe. But more recent studies demonstrated high diversity in South America. In the present project we have conducted an intensive study of the population diversity of T. gondii and surveyed the extent of genetic variation among natural T. gondii isolates on a global scale in order to better understand the population dynamics and pathogenesis of this parasite. To this …

Kinetic, Mechanistic, And Structural Investigation Of Features Controlling Stereoselectivity Of (R)- And (S)-Hydroxypropyl Com Dehydrogenases From Xanthobacter Autrophicus Strain Py2, Dariusz Adam Sliwa

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Enantiopure alcohols are valuable intermediates in fine organic synthesis, in particular for preparation of biologically active compounds. The necessity of preparing single enantiomer drugs in an optically pure form has triggered much research, especially in the pharmaceutical industry. The biocatalytical production of chiral alcohols by alcohol dehydrogenase enzymes is characterized by the asymmetric reduction of the corresponding ketones, usually with high degree of stereoselectivity. The commercial value of the enzymes as stereoselective biocatalysts has been a significant driving force in understanding features that control their mechanism of catalysis and stereoselectivity. This work focuses on two enantiocomplementary dehydrogenase enzymes ((R …

Maporal Hantavirus Β-Integrin Utilization And Sensitivity To Favipiravir, Kristin K. Buys

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Hantaviruses are members of the Bunyaviridae family of viruses. Pathogenic hantaviruses are the etiologic agents of hemorrhagic fever with renal syndrome (HFRS), a disease principally endemic in the Old World, and hantavirus pulmonary syndrome (HPS), a disease primarily restricted to the Americas. Maporal virus (MPRLV), a recently isolated hantavirus, has been found to cause disease in hamsters that resembles HPS in humans. However, the virus has not been linked to human cases of HPS. Considerable evidence suggests that β-integrin usage mediating infection may serve to distinguish hantaviruses pathogenic to humans from nonpathogenic, but this receptor usage pattern information is not …

Vertical Transmission Of West Nile Virus In Culex Spp. Mosquitoes Of Clark County, Nevada, Vivek K. Raman

UNLV Theses, Dissertations, Professional Papers, and Capstones

West Nile Virus (WNV) is an RNA arbovirus that cycles between mosquitoes and birds, but also infects and causes disease in humans, horses and other vertebrate species. Although most infections are asymptomatic, WNV has emerged as the most common cause of epidemic meningoencephalitis in North America and the leading cause of arboviral encephalitis in the United States.

Mosquitoes of the genus Culex are the most active vectors of WNV in North America. Approximately 85% of human WNV infections in the United States occur in late summer with a peak number of cases in August and September. Vertical transmission is defined …

Two-Phase Partitioning System Using Elvax 40w Polymer For The Biodegradation Of Aqueous Phenols, Amit Suresh Ghode

Masters Theses & Specialist Projects

A solid-liquid two phase partitioning system (TPPS) is a new technology platform for destroying toxic organic compounds. TPPS have traditionally been operated by using an immiscible organic phase which partitions organic compounds into the aqueous phase. TPPS using an immiscible organic phase suffers from several limitations such as the organic phase could be biodegradable and hence only certain compatible microbial strains could be used. This therefore, eliminates the desired use of mixed microbial populations for efficient degradation. A solid-liquid two phase partitioning system, in which solid polymeric beads replace liquid organic phase, appears to have benefits over the traditional liquid-liquid …