Biochemistry, Biophysics, and Structural Biology Commons^™

Biochemical Characterization Of Fsa1572 From Fervidibacter Sacchari, The First Hyperthermophilic Gh50 With Β-1, 4-Glucanase Activity, Jonathan Covington

UNLV Theses, Dissertations, Professional Papers, and Capstones

The bacterium Fervidibacter sacchari is an aerobic hyperthermophile that catabolizes various polysaccharides and is the only cultivated member of the class Fervidibacteria within the phylum Armatimonadota. Among its glycoside hydrolase (GH) cache is an enzyme from GH family 50 (GH50), an understudied family with only 25 characterized representatives and two known activities from 1,518 predicted members in the Carbohydrate-Active EnZyme (CAZy) Database. Here, we expressed, purified, and functionally characterized an extracellular GH50 from F. sacchari called Fsa1572. Using colorimetric assays, we show it has novel β-1,4-glucanase activity and only weak agarose activity that is typical for GH50 enzymes. The purified …

Poop! There It Is! Anti-Germinants And Biological Variables As Modulators Of Clostridioides Difficile Infection (Cdi), Jacqueline Renee Phan

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridioides difficile infection (CDI) is a major cause of antibiotic-associated diarrhea. Due to its insidious nature, the Centers for Disease Control and Prevention (CDC) has declared CDI an urgent threat. A key characteristic of C. difficile is its ability to form dormant spores that act as the infectious vehicles for disease. In the gut, spores recognize bile salts to germinate into toxin-producing cells.Dysbiosis of the gut microbiome is a key factor in allowing the C. difficile spores to germinate. Normal gut microbiota naturally protects from CDI. However, biological variables such as diet and sex have been found to modulate to …

Transcriptional Control Of Virulence Genes In The Bacterial Pathogen Shigella Flexneri, Joy Asami Mckenna

UNLV Theses, Dissertations, Professional Papers, and Capstones

Shigella species, the causal agents of bacillary dysentery, use a type III secretion system (T3SS) to inject two waves of virulence proteins, known as effectors, into the human colonic epithelium to subvert host cell machinery. Transcriptional regulation of these virulence genes is controlled by the three-tiered VirF/VirB/MxiE signaling cascade. Of these, VirB has the largest regulon (~50 genes); however, VirB regulatory properties are poorly understood. To date, VirB is known to function to counter or 'anti-silence' transcriptional silencing mediated by the histone-like nucleoid structuring protein H-NS. To better understand VirB regulation, my colleagues and I chose to better define the …

Rapid Evolution Of Starvation Resistance In Drosophila: Physiological And Molecular Mechanisms, Austin Joseph Mckenna

UNLV Theses, Dissertations, Professional Papers, and Capstones

The Gibbs lab has maintained starvation-selected Drosophila melanogaster for >130 generations. These starvation-selected flies evolved an obese phenotype with a suite of physiological differences compared to control-fed flies. Previous studies have shown that long-term starvation-selected Drosophila contain more lipids, have lower metabolic rates and develop more slowly than controls. This dissertation encompasses 1) Examining the molecular mechanisms contributing to starvation resistance, 2) Functional validation of the candidate plin1 allele, and 3) Rapid physiological and genomic evolution in starvation-selected Drosophila.

Starvation-selected Drosophila survive starvation conditions much longer than control-fed flies. This study took a simple approach to identify possible differences in …

Aligning Best Practices In Student Success And Career Preparedness: An Exploratory Study To Establish Pathways To Stem Careers For Undergraduate Minority Students, Kimberly D. Kendricks, Anthony A. Arment, K. V. Nedunuri, Cadance A. Lowell

Journal of Research in Technical Careers

Undergraduate minority retention and graduation rates in STEM disciplines is a nationally recognized challenge for workforce growth and diversification. The Benjamin Banneker Scholars Program (BBSP) was a five-year undergraduate study developed to increase minority student retention and graduation rates at an HBCU. The program structure utilized a family model as a vehicle to orient students to the demands of college. Program activities integrated best K-12 practices and workforce skillsets to increase academic preparedness and career readiness. Findings revealed that a familial atmosphere improved academic performance, increased undergraduate research, and generated positive perceptions of faculty mentoring. Retention rates among BBSP participants …

Effect Of Bmp Treatment On Periostin Gene Expression In Pre-Osteoblastic Mc3t3-E1 Mouse Cells, Vincent Lee Khang

UNLV Theses, Dissertations, Professional Papers, and Capstones

Periostin is a secreted, extracellular matrix (ECM) protein widely expressed within collagen-rich fibrous connective tissues of the body including the periodontal ligament (PDL), bone, skin, heart, and cornea. Periostin has been shown to serve many important regulatory functions including cell adhesion, cell motility, wound healing and of particular importance to the dental field, differentiation of osteoblasts. The deletion of periostin compromises osteoblast attachment to bone matrix and induces a reduction in mineralization and expression of bone markers, including type I collagen, osteocalcin, osteopontin and alkaline phosphatase. Periostin has also been shown to play a significant role in collagen fibrillogenesis by …

The Caspase Cascade During Hibernation In The Golden-Mantled Ground Squirrel, Spermophilus Lateralis, Michael David Treat

UNLV Theses, Dissertations, Professional Papers, and Capstones

In several human pathologies like heart attack, stroke, neurodegenerative diseases, and autoimmune disorders, widespread cell death, or apoptosis, is a major cause of organ dysfunction and death. Hibernating golden-mantled ground squirrels, Spermophilus lateralis, experience numerous conditions during the winter that are known to be pro-apoptotic in other mammal systems (e.g. extreme hypothermia, ischemia and reperfusion, acidosis, increased reactive oxygen species, bone and muscle disuse). However, studies suggest that hibernators may invoke a protective phenotype to limit widespread cell damage and loss during the hibernation season. Could regulating apoptosis provide protection against the harmful conditions experienced during the hibernation season? Could …

Software For Sequence Analysis Of Variants In Functional Screening Libraries And Personalized Genome Files, Jacklyn Michelle Newsome

UNLV Theses, Dissertations, Professional Papers, and Capstones

Detailed knowledge of protein function is critical for both the study of protein interactions and the development of drugs which target specific proteins. Currently, there are few techniques that directly examine protein function. The techniques that are available are time consuming and can only address one variant of a protein at a time. Our laboratory has designed 3 high throughput protein function screens. We hypothesize that these will address this shortfall.

The first screen is the Chimeric Minimotif Decoy (CMD) Assay. For this screen, we constructed red fluorescent proteins with one or more C-terminal minimotifs. Minimotifs are short, contiguous amino …

A Potential Solution To A Poopy Problem: Bile Salt Analogs As Prophylactics Against Clostridium Difficile Infection, Jacqueline Renee Phan

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridium difficile infection (CDI) is a major cause of antibiotic-associated diarrhea. In 2011, over 500,000 patients were diagnosed with CDI in the United States and over 29,000 people died of CDI-related complications. With an average of $35,000 to treat a single case of inpatient CDI, cost burden to the healthcare system can reach up to $3.2 billion annually. As both hospital- and community-acquired CDI incidences rise due to the emergence of hypervirulent strains and CDI reoccurrences of up to 25%, standard treatments are rendered less effective and new methods of prevention are critical.

CDI is caused by bacteria called Clostridium …

Paenibacillus Larvae Spore Germination And American Foulbrood Disease Development In Honey Bee Larvae, Israel Alvarado

UNLV Theses, Dissertations, Professional Papers, and Capstones

Honey bee (Apis mellifera) pollination plays an important role in meeting our agricultural needs, yet we are incapable of completely preventing a decline in bee health, partially due to bacterial diseases. American Foulbrood (AFB), a bacterial disease of honey bee larvae, is particularly troublesome because the infectious agent is the bacterial spore of Paenibacillus larvae. Bacterial spores are resistant to extreme temperatures, unaffected by antibiotics, withstand exposure to disinfectants, and can remain dormant for years until they can revert back to vegetative cells.

Our research investigated P. larvae spore germination at the functional and pathogen-host level. We have found that …

Secondary Structure, A Missing Component Of Sequence- Based Minimotif Definitions, David P. Sargeant, Michael R. Gryk, Mark W. Maciejewsk, Vishal Thapar, Vamsi Kundeti, Sanguthevar Rajasekaran, Pedro Romero, Keith Dunker, Shun-Cheng Li, Tomonori Kaneko, Martin Schiller

Life Sciences Faculty Research

Minimotifs are short contiguous segments of proteins that have a known biological function. The hundreds of thousands of minimotifs discovered thus far are an important part of the theoretical understanding of the specificity of protein-protein interactions, posttranslational modifications, and signal transduction that occur in cells. However, a longstanding problem is that the different abstractions of the sequence definitions do not accurately capture the specificity, despite decades of effort by many labs. We present evidence that structure is an essential component of minimotif specificity, yet is not used in minimotif definitions. Our analysis of several known minimotifs as case studies, analysis …

A Genetic, Transgenic, And Transcriptomic Analysis Of Larval Salivary Gland Physiology In Drosophila Melanogaster, Elana A. Paladino

UNLV Theses, Dissertations, Professional Papers, and Capstones

Cholesterol is the precursor to a unique class of lipophilic signaling molecules called steroid hormones that initiate the development of sexual characteristics, reactions to stress, and maintenance of metabolism, among many other functions. Although much progress has been made in understanding the function of these signaling hormones, we do not fully understand how a single steroid can cause many distinct, tissue-specific responses. Drosophila melanogaster is an effective model for understanding steroid hormone action because of its simplicity. The steroid molting hormone 20-hydroxyecdysone (hereafter, 20E) is the primary active steroid in Drosophila and mediates not only larval molts and the emergence …

Achieving High Accuracy Prediction Of Minimotifs, Tian Mi, Sanguthevar Rajasekaran, Jerlin Camilus Merlin, Michael R. Gryk, Martin Schiller

Life Sciences Faculty Research

The low complexity of minimotif patterns results in a high false-positive prediction rate, hampering protein function prediction. A multi-filter algorithm, trained and tested on a linear regression model, support vector machine model, and neural network model, using a large dataset of verified minimotifs, vastly improves minimotif prediction accuracy while generating few false positives. An optimal threshold for the best accuracy reaches an overall accuracy above 90%, while a stringent threshold for the best specificity generates less than 1% false positives or even no false positives and still produces more than 90% true positives for the linear regression and neural network …

Hivtoolbox, An Integrated Web Application For Investigating Hiv, David P. Sargeant, Sandeep Deverasetty, Yang Luo, Angel Villahoz Baleta, Stephanie Zobrist, Viraj Rathnayake, Jacqueline C. Russo, Jay Vyas, Mark A. Muesing, Martin Schiller

Life Sciences Faculty Research

Many bioinformatic databases and applications focus on a limited domain of knowledge federating links to information in other databases. This segregated data structure likely limits our ability to investigate and understand complex biological systems. To facilitate research, therefore, we have built HIVToolbox, which integrates much of the knowledge about HIV proteins and allows virologists and structural biologists to access sequence, structure, and functional relationships in an intuitive web application. HIV-1 integrase protein was used as a case study to show the utility of this application. We show how data integration facilitates identification of new questions and hypotheses much more rapid …

Molecular Evolution And Historical Biogeography Of New World Birds, Brian T. Smith

UNLV Theses, Dissertations, Professional Papers, and Capstones

Deciphering the patterns of how biodiversity has evolved across time and space has remained a fundamental objective for biologists for the last 200 years. Researchers are faced with the challenge of interpreting the complexity of evolutionary patterns that have been generated over the deep history of the Earth. The advancement of DNA sequencing technology has yielded a new and powerful genetic toolkit that has allowed biologists to address novel evolutionary questions. For my dissertation research, I used molecular genetics and a statistical framework to study the evolution and historical biogeography of birds distributed in North and South America. My dissertation …

Partitioning Of Minimotifs Based On Function With Improved Prediction Accuracy, Sanguthevar Rajasekaran, Tian Mi, Jerlin Camilus Merlin, Aaron Oommen, Patrick R. Gradie, Martin R. Schiller

Life Sciences Faculty Research

Background

Minimotifs are short contiguous peptide sequences in proteins that are known to have a function in at least one other protein. One of the principal limitations in minimotif prediction is that false positives limit the usefulness of this approach. As a step toward resolving this problem we have built, implemented, and tested a new data-driven algorithm that reduces false-positive predictions.

Methodology/Principal Findings

Certain domains and minimotifs are known to be strongly associated with a known cellular process or molecular function. Therefore, we hypothesized that by restricting minimotif predictions to those where the minimotif containing protein and target protein have …

From The Valleys To The Mountains: The Biographic History Of Antelope Squirrels, Bats, And Chipmunks In Western North America, Stacy James Mantooth

UNLV Theses, Dissertations, Professional Papers, and Capstones

Genetic differentiation within and between species often coincides with significant geological or climatic changes that have shaped the sizes and locations of their geographic ranges and altered the connectivity between populations over time. Across western North America, many endemic taxa experienced high levels of initial divergence associated with geological transformations of the Neogene, with subsequent diversification and geographic structuring of populations associated with climatic changes during the Quaternary. As such, we can use a combination of molecular markers and genetic analyses to effectively examine the evolutionary and biogeographic histories of populations, species, and regional biotas whose signatures of differentiation are …

Venn, A Tool For Titrating Sequence Conservation Onto Protein Structures, Jay Vyas, Michael R. Gryk, Martin R. Schiller

Life Sciences Faculty Research

Residue conservation is an important, established method for inferring protein function, modularity and specificity. It is important to recognize that it is the 3D spatial orientation of residues that drives sequence conservation. Considering this, we have built a new computational tool, VENN that allows researchers to interactively and graphically titrate sequence homology onto surface representations of protein structures. Our proposed titration strategies reveal critical details that are not readily identified using other existing tools. Analyses of a bZIP transcription factor and receptor recognition of Fibroblast Growth Factor using VENN revealed key specificity determinants. Weblink: http://sbtools.uchc.edu/venn/.

A Proposed Syntax For Minimotif Semantics, Version 1., Jay Vyas, Ronald J. Nowling, Mark W. Maciejewski, Sanguthevar Rajasekaran, Michael R. Gryk, Martin R. Schiller

Life Sciences Faculty Research

BACKGROUND:

One of the most important developments in bioinformatics over the past few decades has been the observation that short linear peptide sequences (minimotifs) mediate many classes of cellular functions such as protein-protein interactions, molecular trafficking and post-translational modifications. As both the creators and curators of a database which catalogues minimotifs, Minimotif Miner, the authors have a unique perspective on the commonalities of the many functional roles of minimotifs. There is an obvious usefulness in standardizing functional annotations both in allowing for the facile exchange of data between various bioinformatics resources, as well as the internal clustering of sets of …

Minimotif Miner 2nd Release: A Database And Web System For Motif Search, Sanguthevar Rajasekaran, Sudha Balla, Patrick R. Gradie, Michael R. Gryk, Krishna Kadaveru, Vamsi Kundeti, Mark W. Maciejewski, Tian Mi, Nicholas Rubino, Jay Vyas, Martin R. Schiller

Life Sciences Faculty Research

Minimotif Miner (MnM) consists of a minimotif database and a web-based application that enables prediction of motif-based functions in user-supplied protein queries. We have revised MnM by expanding the database more than 10-fold to approximately 5000 motifs and standardized the motif function definitions. The web-application user interface has been redeveloped with new features including improved navigation, screencast-driven help, support for alias names and expanded SNP analysis. A sample analysis of prion shows how MnM 2 can be used.

Las Vegas Wash Water Quality Monitoring Program: 1996 Report Of Findings, Richard A. Roline, James J. Sartoris, U.S. Bureau Of Reclamation, U.S. Geological Survey

Publications (WR)

Las Vegas Wash, a natural wash east of the city of Las Vegas, Nevada, carries stormwater, groundwater drainage, and sewage effluent from three sewage treatment plants to Lake Mead. The Wash provides nearly the only surface water outlet for the entire 2,193 mi² of Las Vegas Valley. A drainage area of 1,586 mi² contributes directly to the Wash through surface flow which is channeled to Las Vegas Bay of Lake Mead, while drainage of the remaining 607 mi² is presumably subsurface and may drain toward Las Vegas Wash.

In the 1930's and 1940's, sewage treatment plants were …

Study Of Existing Information Concerning Water Quality Within Lake Mead, Vicki Scharnhorst, Southern Nevada Water Authority

Publications (WR)

The purpose of Task 010A15M of the Southern Nevada Water Authority (SNWA) Treatment and Transmission Facility (TTF) contract is to conduct a study of existing information concerning water quality within Lake Mead and identify additional water quality studies that are needed to supplement existing data.

The objective of this task is not to discuss treatability of the raw water source; this is addressed by Task 010A18M, "Define Water Treatment Requirements." In addition, a narrative on the effect of pending Safe Drinking Water Act amendments and a determination of treated water quality goals is included in Task 010A16M, "Review Safe Drinking …

Limnological Monitoring Data For Lake Mead During 1988, Suzanne E. Leavitt, Larry J. Paulson, State Of Nevada: Division Of Environmental Protection

Publications (WR)

Limnological monitoring was conducted in Las Vegas Bay and Boulder Basin from April to December of 1988. The purpose of the monitoring was to (i) document possible changes in water quality resulting from decreased phosphorus loading and increased ammonia in Las Vegas Wash, and (ii) establish a data base for evaluating the adequacy of water quality standards.

Characterization Of The Aquatic Environment In Lake Mead Near The Proposed Spring Canyon Pumped-Storage Project, And Assessment Of Potential Aquatic Impacts, Charles R. Liston, Stephen J. Grabowski, Bureau Of Reclamation

Publications (WR)

A pumped storage system consists of an upper reservoir and lower reservoir separated by an elevation difference. During low demand energy periods such as nights and weekends water is pumped from the lower to the upper reservoir using available energy from conventional steam electric power plants. During high energy demand periods, such as mornings and afternoons of weekdays, upper reservoir water is allowed to drop back down through the same system of water conduits and turbines, generating electricity to conveniently meet abrupt electrical energy requirements. The same water turbines thus act both as pumps and as conventional hydroelectric turbines.

Because …

Limnological Monitoring Data For Lake Mead During 1987: Technical Report No. 20, Larry J. Paulson

Publications (WR)

Limnological monitoring was conducted in Las Vegas Bay and Boulder Basin from April to December of 1987. The purpose of the monitoring was to (i) document possible changes in water quality resulting from decreased phosphorus loading in Las Vegas Wash, and (ii) establish a data base for evaluating the adequacy of water quality standards.

Changes In The Morphometry Of Las Vegas Wash And The Impact On Water Quality, Richard A. Roline, James J. Sartoris, U.S. Bureau Of Reclamation

Publications (WR)

Las Vegas Wash, a natural wash east of Las Vegas, Nevada, carries stormwater, groundwater drainage, and sewage effluent from two sewage treatment plants to Lake Mead. Over 80 percent of the normal discharge of approximately 3.4 m³/s (120 ft³/s) consists of effluent from the City of Las Vegas and Clark County sewage treatment plants. Beginning in the 1950s, a large wetland area developed along the wash that supported waterfowl populations and contributed to some water quality transformations. Heavy rains and subsequent flooding in the area in 1983 and 1984 resulted in erosion and channelization that greatly …

Lake Mead Cover Enhancement Project, Jennifer Stevens Haley, Suzanne Leavitt, Larry Paulson, Donald H. Baepler

Publications (WR)

Ninety-three wildlife agencies were surveyed for information on their attempts to improve fish habitat. In addition, an annotated bibliography including over 100 summaries was completed on:

1. largemouth bass cover requirements and preferences,

2. use and effectiveness of artificial cover,

3. aquatic plant introduction and species requirements for germination and establishment,

4. terrestrial plant introduction and species requirements for germination and establishment, and

5. nutrient exchange between sediment, aquatic plants, and water.

A reconnaissance of existing terrestrial and aquatic vegetation was completed in June 1986 including the production of a video tape of the Nevada shoreline of Lake Mead.

Cover …

Las Vegas Wash Advanced Water Quality Study: Final Report, Richard A. Roline, James J. Sartoris, U.S. Bureau Of Reclamation

Publications (WR)

The purpose of the Las Vegas Wash Advanced Water Quality Study is to determine the existence, extent, and mechanisms of nutrient and toxin stripping in Las Vegas Wash under present conditions and under future conditions, both with and without construction of the proposed salinity control unit as described by the Bureau of Reclamation (USBR, 1982b). This study was performed for the Lower Colorado Region Division of Planning by personnel of the Environmental Sciences Section of the Division of Research and Laboratory Services, E&R Center, Denver, Colorado. Work on the study began in February 1983.

The general approach adopted for this …

Historical Patterns Of Phytoplankton Productivity In Lake Mead, Richard T. Prentki, Larry J. Paulson

Publications (WR)

Lake Mead was impounded in 1935 by the construction of Hoover Dam. The Colorado River was unregulated prior to then and therefore was subjected to extreme variations in flows and suspended sediment loads. Hoover Dam stabilized flows and reduced suspended sediment loads downstream, but Lake Mead still received silt-laden inflows from the upper Colorado River Basin. The Colorado River contributed 97% of the suspended sediment inputs to Lake Mead, and up to 140 x 1O⁶ metric tons (t) entered the reservoir in years of high runoff. Most of the sediments were deposited in the river channel and formed an …

Effects Of Wastewater Discharges On Periphyton Growth In Lake Mead, Nevada-Arizona, Marsha Korb Morris

Publications (WR)

A study of the effects of secondary-treated wastewater on periphyton growth in Lake Mead, Nevada-Arizona was conducted from September 1979 to December 1980. Periphyton ash-free dry weight, chlorophyll-a, dominant species composition, and alkaline phosphatase activity were measured on fiberglass substrates. Substrates were incubated for two to four weeks in littoral and limnetic habitats. Physical and chemical variables and phytoplankton chlorophyll-a were measured concurrently.

Transparency increased with increasing distance from the discharge. Secchi depth ranged from 0.75 m at the discharge confluence (station 2) in August, to greater than 20 m at the most distant stations (stations 9 and 10) in …