Biochemistry Commons^™

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 …

Beryllium Is A Potent And Unique Gsk-3Β Inhibitor With Potential To Differentially Regulate Glycogen Synthase And Β-Catenin, Ata Ur Rahman Mohammed Abdul

UNLV Theses, Dissertations, Professional Papers, and Capstones

Glycogen Synthase Kinase-3β (GSK-3β) is an important serine/threonine kinase that phosphorylates/regulates diverse and metabolically important proteins. Some of the important substrates of GSK-3β are glycogen synthase, tau, β-catenin, cyclin D1, axin, c-jun, c-myc, Heat Shock Factor–1, BCL-3, CREB, Histone H1.5, mdm2, p21 (CIP1), pyruvate dehydrogenase and many more. De-regulation of GSK-3β has been implicated in diseases like cancer, Alzheimer’s disease, bipolar disorder and type 2 diabetes mellitus. Currently, GSK-3β is one of the most widely studied proteins and there is a great interest in developing potent and efficient GSK-3β-inhibitors for research as well as therapeutic purposes. We demonstrate that beryllium …

Alternate Germinants Of C. Difficile, A Leading Hospital Pathogen, Sean Tanzey, Ernesto Abel-Santos

McNair Poster Presentations

Clostridium difficile infections (CDI) are the leading nosocomial infections worldwide. Humans are asymptomatic carriers of C. difficile spores in the intestinal tract. The process known as germination occurs when otherwise harmless C. difficile spores are converted to toxin-producing cells upon recognition of bile salts in humans. This distinctive transition ultimately leads to the onset of disease and recurrent CDI. Germination profiles will be characterized in response to peptidoglycan (PG) fragments isolated from various bacterial species. These specific peptidoglycan fragments contain different amino acid residues that may induce different germination responses. Purification and structural determination of the peptidoglycan fragments will be …

Synthesis And Characterization Of Novel Platinum(Ii) And Platinum(Iv) Complexes Containing 4,4′--Disubstituted--2,2′--Bipyridine Ligands For The Treatment Of Cancer, Van Vo

UNLV Theses, Dissertations, Professional Papers, and Capstones

Three series of platinum(II) and platinum(IV) complexes containing 4,4′-disubstituted-2,2′-bipyridine ligands have been synthesized and characterized by 1H NMR, 13C NMR spectroscopy, elemental analysis, mass spectroscopy, and differential scanning calorimetry measurements. The MTS cell proliferation assay was used to examine the in vitro anti-proliferative activities of these complexes in various human breast, lung, and prostate cancer cells. The cell's response to the complexes varies between different cell lines; however, the low EC₅₀ values determined from the MTS data indicate that several of the complexes are much more potent than cisplatin.

Flow cytometric analysis of selected compounds revealed induction of apoptosis …

Initial Characterization Of A Conserved Active Site Residue For The Cdc34 Ubiquitin Conjugating Enzyme, Arvin Akoopie

Honors College Theses

Ubiquitin-conjugating enzymes (E2s) covalently modify protein substrates with ubiquitins. The active site cysteine residues on E2s are essential for catalyzing the transfer of ubiquitin from the E2 active site onto the protein substrate, however there is a limited amount of information available concerning additional active site residues for E2s that may also participate in catalysis. Cdc34 is an essential E2 that has merited the lion’s share of attention for biochemical analysis of the E2 family. Previous phylogenetic analysis of Cdc34 amino acid sequences has identified an invariably conserved histidine residue close to the active site cysteine in the primary structure, …

Synthesis And Characterization Of Pt(Ii) Complexes For Anticancer Therapy, Mihaela A. Ciulei, Pradip K. Bhowmik

McNair Poster Presentations

The first platinum-based drug was discovered and approved by Food and Drug Administration (FDA) in 1978 is cis-diamminedichloroplatinum (II) (cisplatin or CDDP). Cisplatin is used for about 50% of the chemotherapeutic cancer treatments along with its two analogues carboplatin and oxaliplatin. So far these drugs have been used extensively as treatment for ovarian, bladder, head and neck, and lung cancers. Although cisplatin has been used so often, it has toxic side effects and drug resistance.1-4 Due to these limitations other compounds have been synthesized. Specifically, our lab in conjunction with a biochemistry lab has recently published one article …

What Can Biochemistry Students Learn About Protein Translation? Using Variation Theory To Explore The Space Of Learning Created By Some Common External Representations, Thomas J. Bussey

UNLV Theses, Dissertations, Professional Papers, and Capstones

Biochemistry education relies heavily on students' ability to visualize abstract cellular and molecular processes, mechanisms, and components. As such, biochemistry educators often turn to external representations to provide tangible, working models from which students' internal representations (mental models) can be constructed, evaluated, and revised. However, prior research has shown that, while potentially beneficial, external representations can also lead to alternative student conceptions.

Considering the breadth of biochemical phenomena, protein translation has been identified as an essential biochemical process and can subsequently be considered a fundamental concept for biochemistry students to learn. External representations of translation range from static diagrams to …

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 …

Characterization Of Beryllium As A Novel Agent To Study Cell Cycle Arrest And Cellular Senescence, Priyatham Gorjala

UNLV Theses, Dissertations, Professional Papers, and Capstones

Cancer cells evade senescence, apoptosis, and other constraints on proliferation, often via mutation of the p53 tumor suppressor gene (TP53). Normal human lung fibroblasts have been shown to enter premature senescence upon exposure to beryllium. In these cells, BeSO₄ stabilizes p53 protein, increases p21 gene expression, induces senescence-associated β-galactosidase activity and causes cell proliferation arrest. In the present study, we have investigated whether BeSO₄ is able to induce similar effects in cancer cells that have wildtype p53. We have demonstrated that beryllium salt at low concentration can induce molecular changes in the p53 signaling pathway leading to cell …

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 …

Anti-Germinants As A New Strategy To Prevent Clostridium Difficile Infections, Amber Janece Howerton

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridium difficileinfections (CDI) have emerged as a leading cause of hospital-associated complications. CDI is the major cause of antibiotic-related cases of diarrhea and nearly all cases of pseudomembranous colitis. The infective form of C. difficileis the spore, a dormant and hardy structure that forms under stress. Germination of C. difficile spores into toxin producing bacteria in the GI tract of susceptible patients is the first step in CDI establishment. Patient susceptibility occurs with a disruption of the natural gut microbiota by broad-spectrum antibiotics. Antibiotic treatments usually resolve CDI but refractory cases are on the rise. Of great concern is the …

The Role Of A Transcription Factor In Regulating Rice Response To Drought Stress, Diana Ha, Liyuan A. Zhang, Jeffery Shen

Undergraduate Research Opportunities Program (UROP)

The current water shortage is a major concern in regard to our global climate change crisis. A decrease in the availability of water will have direct effects on the development of plants. Some crops, such as Oryza sativa, or commonly known as rice, requires an abundant amount of water for adequate growth. With the water shortage crisis, it will become extremely difficult to harvest such crops to meet the world’s food demand. However, many plants have evolved mechanisms for overcoming and tolerating stresses such as drought. My research focuses on studying the proteins involved with these mechanisms. The WRKY superfamily …

Ubiquitylation Of Proteins In The Frozen Wood Frog, Alexa Khan, Michael Ulrich, Kenneth Storey, Frank Van Breukelen

Undergraduate Research Opportunities Program (UROP)

Wood frogs (Rana sylvatica) are able to withstand freezing. Respiratory and cardiac activity ceases when frozen. Homeostatic functions like protein synthesis and degradation presumably must also be compromised. We investigated the fate of ubiquitin-dependent proteolysis in the freeze-thaw cycle and how that might gives clues to wood frog survival. We performed western blots for ubiquitin conjugates

Novel Thermophilic Cellulolytic Isolates Belonging To The Phylum Chloroflexi, Maryknoll Palisoc, Jessica K. Guy, Joseph P. Peacock, Duy C. Trinh, Jeremy A. Dodsworth, Brian P. Hedlund

Undergraduate Research Opportunities Program (UROP)

Current biofuel technologies utilize valuable foodstuffs, such as corn kernels and cane sugar, as sources of easily metabolized sugars. Microbes are used to ferment these sugars into bioethanol, a first-generation biofuel. However, in order to avoid diverting foodstuffs from the food supply, the development of second-generation biofuels technology is necessary. Second-generation biofuels are produced by converting structurally complex lignocellulosic biomass, such as agricultural and municipal wastes, to fermentable sugars or directly to biofuels.

The major technological hurdle limiting the mass production of second-generation biofuels is the difficulty in efficiently converting structurally complex lignocellulosic materials to fermentable sugars or directly to …

Inactivation Of Spo0a Gene Increases Stationary Phase Mutagenesis In Bacillus Subtilis, Denisse Reyes, Amanda Prisbrey, Holly Martin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

Stationary phase mutagenesis occurs when a population of cells acquires mutations conferring escape from nongrowing or stress conditions. This type of mutations is observed in nutritionally starved cells. Because the mutations occur after the onset of stress and in cells that are in non-replicative conditions, elucidating the underlying mechanisms contributes novel views to the process of evolution and apply to the formation of cancer in human cells and antibiotic resistance in microbial pathogens. Studies have shown that in Bacillus subtilis, the Mfd protein which is a transcription repair coupling factor is necessary for this phenomenon to occur. Here, we investigate …

Synthesis Of Chimeric Receptors Essential For Spore Germination, Christopher Yip, Christian Ross, Eduardo Robleto, Ernesto Abel-Santos

Undergraduate Research Opportunities Program (UROP)

Various species of bacteria have been reported to form an endospore, a metabolically dormant cell, during times of nutrient deficiencies and extreme stress. These said structures are outstandingly resistant to harsh chemicals, extreme temperatures, and can revert back to a metabolically active cell, through a process known as germination, when the necessary conditions are met. The rigid membrane of the endospore contains various germination (Ger) receptors which sense the external environment for necessary metabolites and germinants. Ger receptors are encoded by tricistronic operons that produce three distinct membrane proteins, the A, B, and C subunits. Although the function of the …

Investigating The Origin Of Coprolites From Three Great Basin Caves, Chelsey Vandrisse, Duane P. Moser, David Rhode

Undergraduate Research Opportunities Program (UROP)

The study of coprolites (mummified feces) is a relatively new endeavor, which enables investigations of the health and diet of ancient people and provides some of the oldest evidence to date for the human habitation in North America (2). In this project, 18 coprolites were examined from archeological digs at three Great Basin caves: the Bonneville Estates Rockshelter (UT), Hidden Cave (NV), and Top of the Terrace Rockshelter (UT). The main objectives were: 1) to verify human origin through the presence of mitochondrial DNA (mtDNA) and 2) assuming human origin, characterize intestinal microflora of Native Americans prior to European contact. …

The Role Of An Abc Transporter As A Steroid Antagonist In Drosophila, Gregory King, Andrew Andres

Undergraduate Research Opportunities Program (UROP)

Drosophila melanogaster are holometabolous insects that have several distinct life stages including larvae and a winged adult. The larval stage is mainly a time of feeding and growth, while the adult stage is optimized for sexual reproduction and dissemination. The larval stage can itself be divided into three time periods, or instars: 1st (L1), 2nd (L2), and 3rd (L3) (Figure 1). Larval growth – both between instars and beyond – depends on specific signaling pathways controlled by a cholesterol derived steroid, 20-hydroxyecdysone (20E). Although 20E is a systemic developmental signal, little is known about the molecular details of how different …

Cell Migration Dynamics After Alteration Of Cell-Cell Contacts In Fibrosarcoma And Glioblastoma Cell Lines, Hassan S. Rizvi, Ronald K. Gary

Undergraduate Research Opportunities Program (UROP)

Cell migration is a vital component of metastasis. In this study, our intent was to study cell migration by alteration of the Wnt/GSK-3 Pathway. Since BeSO4 is a known GSK-3 kinase inhibitor, we hypothesized that this agent would cause cell migration to decrease as a result of β-catenin stabilization. Two human cell lines, HT-1080 (fibrosarcoma) and A172 (glioblastoma), were used to observe migration levels in the presence and absence of BeSO4. Our results show that cell migration is diminished for cells that were pre-treated with BeSO4, in comparison to the untreated (control) cells.

Isolation, Characterization, And Genome Sequence Of The First Representative Of A Novel Class Within The Chloroflexi That Is Abundant In Some U.S. Great Basin Hot Springs And May Play Important Roles In N And C Cycling, Jeremy A. Dodsworth, Senthil K. Murugapiran, Jonathan Gevorkian, James Han, Tanja Woyke, Susan M. Lucas, Sam Pitluck, Len Pennacchio, Lynne Goodwin, Brian P. Hedlund

Undergraduate Research Opportunities Program (UROP)

A thermophilic, facultatively microaerophilic, heterotrophic bacterium, designated strain JAD2, was isolated from sediments of Great Boiling Spring (GBS), an ~80oC, circumneutral hot spring in the Great Basin GB). The strain grew anaerobically on yeast extract or peptone with an optimal growth temperature of 70-75oC. Growth was stimulated by addition of 0.01 atm O2 to the culture vessel headspace, but was inhibited by higher concentrations (0.2 atm). Cells of JAD2 formed non-motile filaments ranging from 10 to >300 μm in length, which typically decreased in length during stationary phase. 16S rRNA gene-targeted pyrotag sequencing and clone library data suggest that close …

Dna Secondary Structures And Their Contribution To Mutagenesis In B. Subtilis Stationary Phase Cells, Carmen Vallin, Holly Martin, Christian Ross, Ronald Yasbin, Eduardo Robleto

Undergraduate Research Opportunities Program (UROP)

It is widely known and accepted that the cause of many mutations in cells are generated during the replication process of actively dividing cells, however more recent research has shown that mutations also arise in non growing conditions, a phenomenon known as stationary phase mutagenesis. Much of what is known come from studies in eukaryotic and bacterial models. It has been proposed that in non~growing cells, the process of transcription plays an important role in mutagenesis. We test the hypothesis that DNA secondary structures, formed during transcription, promote mutagenesis. The transcription-generated structures are speculated to be prone to mutations by …

Synthesis Of A Chemical Probe For Identification Of Ger Receptor Proteins Of Bacillus Anthracis, Hyelee Lee

UNLV Theses, Dissertations, Professional Papers, and Capstones

Bacillus anthracis,a gram positive, endospore-forming, rod-shaped, aerobic bacterium, is the causative agent of anthrax. A key step in the pathogenic cycle is the spore germination, where the dormant spore transforms into a metabolically active cell. This allows B. anthracis to proliferate and secrete toxins. It is believed that spore germination is triggered when small molecules, known as germinants, are recognized by germination (Ger) receptor proteins located in the inner membrane of spores. Most Ger proteins are encoded by tricistronic operons, resulting in three distinct products, the A-, B-, C-subunits. Genomic sequencing showed that B. anthracis has seven ger receptor …

Accumulation Of Polonium-210 In Different Species Of Fish In Lake Mead, Suraj Ghevarghese John, Shungmugam Nallaperumal, Vernon Hodge

Festival of Communities: UG Symposium (Posters)

Discovered by Pierre Curie and Marie Skłodowska-Curie in 1898, polonium is a chemical element with an atomic number of 84. This rare naturally occurring radioactive element is chemically similar to bismuth and tellurium, and forms in uranium ores. Polonium-210 is an naturally occurring radioactive element with a half-life of 138.376 days(1). This element is found in trace amounts in most organisms. Our research is focused on the accumulation of polonium-210 in fish that occupy in Lake Mead.

The sample species is chosen based on varying lake stratification (layers of the lake). This gives us a good idea of the difference …

The Release Of Calcium In Bacillus Anthracis Pathogenicity Methods, Natiera Magnuson, Manomita Patra Bhowmik, Maria Elena Reynaga, Ernesto Abel-Santos

Festival of Communities: UG Symposium (Posters)

Anthrax infection starts with germination of Bacillus anthracis spores in macrophages. Some bacteria, including B. anthracis, can sporulate in response to environmental stress, such as starvation. During germination, large concentrations of calcium ions are released from the B. anthracis spore. Calcium ions are hydrophilic secondary messengers, and may therefore interfere with detection of the spore by confusing the cell signaling pathways. We investigated calcium release on infected macrophage viability by replacing the calcium stored in B. anthracis spores for other cations via demineralization/remineralization. It was discovered that calcium ions typically out-performed other cations in germination of B. anthracis.

Characterization Of Ips Pini Ipsdienol Dehydrogenase (Idol Dh), Heidi Pak, Claus Tittiger, William Welch

Festival of Communities: UG Symposium (Posters)

Ipsdienol is an important pheromone component for pine engraver beetle, Ips pini. Ipsdienol is a ten carbon monoterpenoid secondary alcohol and ipsdienone is the corresponding ketone. We are characterizing the activity of recombinant IDOL DH produced in Sf9 (insect) cells. The enzyme has a high stereospecificity: (-) ipsdienol was found to be a substrate while (+)-ipsdienol was neither a substrate nor inhibitor. Closely related monoterpenoids, such as nerol, geraniol, and citral, were neither substrates nor inhibitors. Smaller compounds, such as 2-propanol, also failed to act as an inhibitor or substrate. This indicates the binding site of this enzyme is highly …

Effect Of Temperature And Oxygen Levels On Lactate Production In Palaemonetes Pugio, Robin Schofield, Teresa Mika, Carl Reiber

Festival of Communities: UG Symposium (Posters)

Oxygen delivery rates must keep pace with the increased metabolic rate observed in poikilothermic animals exposed to increased temperature if aerobic metabolism is to be maintained. Physical failure of the oxygen delivery system to meet these demands may be a mechanism of cardiac failure in crustaceans at temperature extremes. As part of the determination of the effect of temperature on the cardio-respiratory system, we are measuring whole animal lactate levels in grass shrimp acclimated to 20°C over temperature ranges in normoxic and hyperoxic conditions. Lactate levels are measured as an indicator of a switch from aerobic to anaerobic metabolism. Animals …

Production, Purification And Crystallization Of Membrane Integrated Multimeric Bax, Adelbert Mark Villoso

UNLV Theses, Dissertations, Professional Papers, and Capstones

Apoptosis, or programmed cell death, is a vital process intimately involved in the embryonic development and homeostatic maintenance of all multicellular organisms. The committing step to apoptosis is regulated by a key protein, Bax, and its ability to integrate and form a pore structure at the outer mitochondrial membrane.

Unfortunately, the molecular details of apoptosis remain largely unclear due to the lack of structural data of integral membrane (IM) Bax. Experimental limitations of membrane protein production have slowed the pursuit of an IM-Bax structure simply because standard protocols for producing recombinant IM-Bax are inefficient in producing adequate quantities of IM-Bax …

Development Of Novel Biomarkers In Cancer: Detection Of Circulating Mir-141 As A Potential Prognostic Marker For Prostate Cancer, Jason Cadaoas Gonzales

UNLV Theses, Dissertations, Professional Papers, and Capstones

Prostate cancer (CAP) is the most common epithelial malignancy and the second leading cause of cancer deaths in American men. The identification of predictive and prognostic biomarkers in CAP patients is critical for improving clinical outcomes. Although the measurement of prostate-specific antigen (PSA) and radiographic studies are clinically approved to predict response to therapy, these tests can oftentimes prove to be inadequate in certain patients. Thus, it is important to discover new biomarkers to improve chances of survivability. We and others have shown that longitudinal measurements of circulating tumor cells (CTC) and lactate dehydrogenase (LDH) may aid in predicting response …

Mimosa: A System For Minimotif Annotation, Jay Vyas, Ronald J. Nowling, Thomas Meusburger, David P. Sargeant, Krishna Kadaveru, Michael R. Gryk, Vamsi Kundeti, Sanguthevar Rajasekaran, Martin Schiller

Life Sciences Faculty Research

BACKGROUND:

Minimotifs are short peptide sequences within one protein, which are recognized by other proteins or molecules. While there are now several minimotif databases, they are incomplete. There are reports of many minimotifs in the primary literature, which have yet to be annotated, while entirely novel minimotifs continue to be published on a weekly basis. Our recently proposed function and sequence syntax for minimotifs enables us to build a general tool that will facilitate structured annotation and management of minimotif data from the biomedical literature.

RESULTS:

We have built the MimoSA application for minimotif annotation. The application supports management of …

Antimicrobial Activity Of D-Lenolate®, Andy Phui

UNLV Theses, Dissertations, Professional Papers, and Capstones

Olive trees are one of the most important fruit trees in the Mediterranean. Although not validated by research, olive leaves are traditionally believed to fight off fever and infections. It has been shown that olive leaf extracts possess antimicrobial activity. Olive leaf extracts contain polyphenols. One of the major phenolic compounds is oleuropein. Oleuropein and other polyphenols have been shown to exhibit antimicrobial activity. East Park Research (EPR) developed an extraction process that they claim does not alter the chemical composition of the olive leaves. The extract is known by the commercial name d-lenolate®. Studies have provided evidence that d-lenolate®, …