Research Projects

Every year students work on a research project led by a UC Davis faculty member. Below is an archive of past projects. Select from the following tabs to explore the project archives.

2018
 

The responses of insect vectors to potyviruses

Student Name: Aaron Lin
UCD Department: Plant Pathology
UCD Mentor: Dr. Clare Casteel

The infection of plants by potyviruses is effects the localization of different proteins like the Nuclear Inclusion a Protease (NIa Pro). In addition, the overall phenotype of the plant can be observed through the effects of the potyvirus on the fecundity of insect vectors on the plant. It has previously been shown that infecting plants with Turnip mosaic virus (TuMV) increases insect attraction and reproduction on host plants. Using previous research that sequenced Turnip mosaic virus (TuMV), Potato virus Y (PVY), and Soybean mosaic virus (SMV) as well as observing their varying effects on fecundity, we were able to identify amino acid sequences of interest within the NIa Pro sequence. We performed fecundity tests on Tobacco etch virus (TEV) and Pepper mottle virus (PepMoV) infected Nicotiana benthamiana to further categorize amino acids within NIa Pro. Tobacco etch virus (TEV) did not significantly increase fecundity, likely due its coinfection with Potato virus Y (PVY), which would increase the fecundity for both of them. Pepper mottle virus (PepMoV) significantly increased fecundity, likely due to its similar homology to Potato virus Y (PVY), which also increases fecundity. The focus of our research was making a 90 amino acid deletion from the N-terminus in Turnip mosaic virus (TuMV)  We observed that the NIa Pro sequence from the mutant TuMV was transported around the cell by vesicles, suggesting that the 90 amino acid deletion from the N-terminus affected the ability of the NIa Pro to relocate to the vacuole.

Contribution of sorghum root aerenchyma to Striga susceptibility

Student Name: Alexander Chen
UCD Department: Plant Biology
UCD Mentor: Dr. Dorota Kawa, Dr. Siobhan Brady

Sorghum bicolor is an essential cereal crop that is extensively parasitized by plants of the genus Striga, resulting in devastating yield loss. This experiment was performed to study patterns of aerenchyma formation in different cultivars of S. bicolor under various conditions, including soil sterility and Striga treatment. Seeds of Striga-resistant genotype SRN39 and Striga-susceptible genotype Shanqui Red (SQR) were germinated in vitro and grown in a greenhouse environment. Images of root cross-sections were electronically processed and quantified based on the amount of aerenchyma formation. An initial analysis on the effects of soil, Striga infection, and soil-Striga infection interaction on aerenchyma development in each cultivar found largely statistically insignificant differences, while a second analysis considering both genotypes found significantly less aerenchyma formation in SRN39 genotype crown and seminal root tips in both natural and sterile soil conditions. In addition, this analysis found that the crown and seminal root tips of S. bicolor grown in sterile soil had significantly less aerenchyma compared to those grown in non-sterile soil. This suggests that the soil microbiome may promote aerenchyma formation in SQR roots and that the near-absence of aerenchyma in SRN39 roots may contribute to its Striga resistance properties. The insight gained through this study can be potentially applied to building a Striga-resistant yet affordable and productive sorghum cultivar.

The Evolutionary History of Tyrosylprotein Sulfotransferases

Student Name: Alvin Zhang
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Grace Rosenquist

Tyrosylprotein sulfotransferases (TPSTs) are responsible for catalyzing the sulfation of peptidyl tyrosine residues. Tyrosine sulfation is a post-translational modification that strengthens a variety of protein-protein interactions, including leukocyte adhesion to endothelial cells and host-pathogen interfacing. We sought to elucidate the evolutionary history of TPST in order to understand the function of sulfation in a wide range of organisms. By comparing the homology of TPSTs across lineages through multiple sequence alignments, the conserved mechanism of substrate recognition and sulfation in both eukaryotes and prokaryotes was elucidated. Binding motifs for the universal sulfate donor 3’-phosphoadenosine 5’-phosphosulfate (PAPS) were found to be conserved in nearly all sulfotransferases, and residues involved in interactions with substrates were found to be highly consistent as well. Molecular visualization software was used to identify trends, as well as nonconserved regions at a level beyond the primary sequence in the catalytic domains of the two isoforms of TPST, TPST-1 and TPST-2. These structural differences may implicate distinct roles for the two isoforms of TPST.

Evaluation of the mitochondrial DNA copy number and deletions in fibroblasts from subjects with the FMR1 premutation

Student Name: Ashwin Kumar
UCD Department: MIND Institute
UCD Mentor: Dr. Cecilia Giulivi

Fragile X syndrome occurs in subjects with >200 CGG repeats in the 5’ untranslated region of the FMR1 gene. Premutation carriers (55-200 CGG repeats) were originally thought to be asymptomatic but some have psychological and metabolic issues among others, and with age some develop the neurodegenerative disease called Fragile X-associated tremor/ataxia syndrome (FXTAS). Studies have been done which have shown that people with FXTAS tend to have mitochondrial functionality problems. However, the exact reason for why this occurs is still elusive. The overall goal of our research was to evaluate the mitochondrial DNA copy numbers (mtDNA) and deletions in fibroblasts from individuals with the FMR1 premutation by qPCR utilizing dual-labeled probes. To measure the mtDNA and deletions, we evaluated the Ct values for each DNA sample provided from Real Time Quantitative-PCR (qPCR).  Our data shows that there was not a significant difference between the mitochondrial DNA copy numbers in fibroblasts from individuals with FXTAS and without it. Our data also shows that non-affected FXTAS carriers had a lower mitochondrial gene ratio, signifying that they had more deletions.

Comparative Genomics of M. stipitatus Cyclase Proteins

Student Name: Brian Lee
UCD Department: Microbiology and Molecular Genetics
UCD Mentor: Dr. Mitchell Singer and Dr. Gaurav Sharma

Cyclases are proteins involved in signal transduction pathways. These proteins help in catalyzing vital intracellular chemical processes such as gene transcription, cellular metabolism, and cardiac function. Myxobacteria such as Myxococcus stipitatus are unique because they display multicellular-like social behaviour in densely grouped populations. Cyclases might be involved in coordinating this type of social behaviour by organizing cell-to-cell communication via various signaling pathways, but the exact mechanism is not yet known. Here, in silico studies were used to identify 14 cyclase proteins in M. stipitatus and explored their architecture, origins, phylogeny, and putative functions. After identification using Pfam domains, the domain architecture of myxobacterial cyclase proteins were compared using different sequence databases. In addition, multiple sequence alignments and phylogenetic trees were used to analyze conserved regions and evolutionary relatedness among the cyclases. This research is focused on understanding the probable evolution and structure of M. stipitatus cyclases, which would be the foundation of further characterization and identification of their unique roles in regulating cell-to-cell communication.

Prediction of Possible Tyrosine Sulfation Sites in Complement Factors

Student Name: Carolyn Lai
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Grace Rosenquist

Three tyrosine sulfation sites are known in complement C4, and sulfation of C4 influences its activity in the complement cascade. Since tyrosine sulfation usually occurs within a group of proteins, this research focuses on whether there are additional tyrosine sulfation sites in complement proteins. The Position-Specific Scoring Matrix (PSSM) reports high scores at tyrosine sites in all of the complement proteins. Some of the complement proteins, such as complement C5, have tyrosine residues located on the exterior of the protein, showing an increased likelihood of becoming sulfated. An increased understanding of tyrosine sulfation contributes to the development of pharmaceutical drugs that could treat diseases related to mutations of complement-mediated immune responses. These diseases include macular degeneration, angioedema, systemic lupus erythematosus, and other complement deficiencies that greatly impact the lives of affected people.

Soil Stabilization and MICP

Student Name: Cat Acuff
UCD Mentor: Dr. Jason T. DeJong & Alex San Pablo

Soil Stabilization and MICP, microscopically induced calcite precipitation, was an engineering focused project in the Department of Civil and Environmental Engineering, under Dr. Jason T. DeJong that included building large scale structures to test soil stability and comparing calcite levels in small scale samples. This research is important in events of earthquakes where liquefaction can easily occur.

Engineering optimal introns

Student Name: Clara Wang
UCD Department: Genome Center
UCD Mentor: Dr. Ian Korf

Background: Intron mediated enhancement (IME) is the phenomenon in eukaryotic organisms in which an intron improves gene expression. Creating new introns that boost gene expression beyond observed introns would be useful in biotechnology applications.
Results: Introns near the promoter are compositionally distinct from those farther down the transcript. This difference in composition is used in an algorithm called the IMEter, which can be used to predict the effectiveness of an intron. Creating artificial introns that simply maximize the IMEter score results in introns that are almost entirely GC. In order to create more natural introns we propose a genetic algorithm, MIA, that simultaneously maximizes IMEter score and the relative entropy compared to average introns. We also propose MIA+, which adds a motif-scoring function to the fitness function.
Conclusions: MIA creates high scoring introns as well as introns with repeated motifs which have previously been shown to be effective in increasing IME. Command line options allow the user to tune the algorithm for IMEter score or motif density. Based on previous experimental data, introns created by MIA are expected to increase gene expression 20-fold or more.

Identification of brighter genetically encoded fluorescent indicators of dopamine and opioids

Student Name: Clara Wu
UCD Department: Biochemistry and Molecular Medicine
UCD Mentor: Dr. Lin Tian

Neurotransmitters and neuropeptides are significant to a number of neurological functions. Understanding how these chemicals act in living cells and animals in real time is made possible with genetically encoded fluorescent indicators that light up neuronal membranes upon binding with their corresponding ligand (trigger chemical). . The goal of our study was to determine if we could further optimize the brightness of our recently developed red dopamine indicator (dLight 1.1 red) and green opioid indicator (KOR 1.1) by altering their genetic codes. After one round of screening, we have determined our current variants to be the highest performers. These sensors show great spatiotemporal resolution when expressed in mammalian cells. These tools provide scientists with a better understanding of dopamine and opioid-related disorders, which will aid in the development of targeted treatments for ADHD, addiction, depression, anxiety, Parkinson’s Disease, schizophrenia, and many others.

Ibuprofen and proteasomal dysfunction in the kidney

Student Name: Colette K. O’Grady
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Aldrin V. Gomes

Ibuprofen is a nonsteroidal anti-inflammatory drug that is used worldwide to inhibit pain and inflammation. Ibuprofen has been shown to have negative side effects in the cardiovascular system, but there has been significantly less research on the side effects in the kidney. The goal of this research is to determine if ibuprofen causes proteasomal dysfunction in the kidney. The proteasome is a complex that is critical for removal of unwanted and oxidized proteins. Proteasome levels were observed through western blotting and proteasome assays, and no significant difference was found in proteasome levels between the ibuprofen and control samples. The proteasome activity was significantly decreased in samples treated with ibuprofen. It was also determined that automated western blot machines are not as effective as manual western blotting methods.

Impacts of Complex Formation Between Chromophoric Dissolved Organic Compounds on UVvisible Absorbance

Student Name: Coral Chen
UCD Department: Land, Air, and Water Resources
UCD Mentor: Dr. Peter Hernes

Absorbance data on chromophoric dissolved organic matter (CDOM) in the UV-visible band is a popular proxy for biomarkers in aquatic systems.  However, these direct correlations between absorbance and other properties of DOM may be flawed if the absorbance data does not accurately reflect the CDOM concentration.  In this study, the impact of synergistic effects between dissolved compounds on CDOM UV-visible absorbance is examined to demonstrate the need to account for these effects when analyzing CDOM absorbance data.  This study looks at various combinations of lignin model compounds, amino acids, and condensed purified tannins, demonstrating that when combined, the absorbance behavior of these compounds is not accurately predicted by Beer’s Law.  The interactions found in this study indicate that any calculations based on CDOM absorbance of water samples should be adjusted for synergistic effects between the compounds present in the sample. In the future, a database of synergistic effects would be important to accurately interpret data from any remote sensing systems that may be developed to monitor DOC in relation to climate change.

Mesoporous Silica Nanocage (MSN) Uptake in Macrophages After Inhalation in MiceMesoporous Silica Nanocage (MSN) Uptake in Macrophages After Inhalation in MiceMesoporous Silica Nanocage (MSN) Uptake in Macrophages After Inhalation in Mice

Student Name: Dana Jung
UCD Department: Center for Health and the Environment
UCD Mentor: Dr. Kent Pinkerton

Mesoporous silica nanocages (MSNs) are inorganic-based nanocarriers containing porous channels. These channels can be filled with medicinal compounds and utilized to target various diseases in the respiratory system. The purpose of this study is to determine the frequency of MSN uptake, retention and fate in macrophages after deposition in the lungs following a single, acute period of inhalation. Macrophages, highly phagocytic cells of the lungs, appear to uptake MSNs at different rates and to different degrees following inhalation into the lungs. The actual MSN uptake by these cells over time was done by calculating the volume of MSN in alveolar macrophages at 1, 7, and 21 days post-inhalation. In previous studies, mice were exposed to MSNs and harvested after 1, 7, and 21 days to show macrophages as the dominant cell type in the lungs to sequester MSN following inhalation. Using cells recovered from the lungs by bronchoalveolar lavage (BAL), as well as examining lung tissues to determine the location of these cells in lungs, MSN with either a positive or negative surface charge to determine its effect on uptake were used to determine the relative frequency of uptake in the macrophages. The results show that the area of MSNs within the macrophages both slightly decrease over time. However, overall it indicates that the MSNs are not clearing out from the lungs or the macrophages.

The Importance of Mutations on Structure and Function of Legionella Pneumophila CMP-N,N’-Diacetyllegionaminic Acid Synthase

Student Name: Echo Tang
UCD Department: Chemistry
UCD Mentor: Dr. John McArthur

Sialic acids are important molecules that are present during cell-to-cell interactions, immunological processes, and pathogenic processes. Their role as markers in pathogens and cancer cells are well understood, but the reason that nature favors some mutations is largely unknown. It has been found that mutations help with sialic acid enzyme substrate binding, specifically in sialic acid synthases that synthesize sialic acids. However, it is difficult to incorporate sialic acid analogs with modifications on the positions C7 to C9 onto glycans using enzymatic methods. The enzyme Legionella Pneumophila CMP-N,N’-Diacetyllegionaminic Acid Synthase (LpCLS), which synthesizes legionaminic acid present in Legionnaire’s Disease, was found to have high structural homology to the human enzyme CMP-sialic acid synthase, N. meningitis CMP-sialic acid synthase (NmCSS). Thermal shift assays also concluded that the stabilizing conditions for LpCLS includes an ideal pH of 8.5 and the presence of the stabilizing ligands CTP and legionaminic acid.

Characterization of one new variant of β-glucosidase B with kinetic and thermal analysis

Student Name: Enya Xing
UCD Department: Genome Center
UCD Mentor: Dr. Justin Siegel

With a deeper knowledge and understanding of the effects of point mutations on amino acid interactions and protein functions, the objective is to create a database to store the collected data and add on to computational learning. Currently, through the FoldIt software, a Rosetta score is provided to serve as a frame of reference for the stability of a mutation. However, only a small amount of designs and mutations engineered achieve the desired function. The database would allow for an increased accuracy and level of knowledge on the prediction on how different point mutations would affect protein stability and efficiency through altered amino acid interactions. The current project focuses on the already well understood β-glucosidase B (BglB) enzyme with a known crystal structure and reports the Michaelis-Menten kinetics and thermal stability of purified mutants. [Insert part about important results]. Through the analysis of the results and the accumulation of data points within the database on BglB, there can be an improved understanding toward point mutations that can be applied to other proteins. The research currently does not have immediate short term results but the data that is collected continues to grow and better the predictive algorithm and protein redesign.

Induction of N immune receptor TIR domain dimerization yields no cell death in N. benthamiana

Student Name: Eric Vuong
UCD Department: Plant Biology and Genome Center
UCD Mentor: Dr. Savithramma P. Dinesh-Kumar

The N resistance gene, which encodes the N protein and confers resistance to tobacco mosaic virus (TMV), is a member of the Toll-interleukin-receptor (TIR) class of plant nucleotide binding leucine rich repeat (NLR) immune receptor . Previously, the N NLR has been shown to dimerize through its TIR domain upon infection with TMV. Here, we attempted to artificially induce dimerization of the N TIR domain in the absence of TMV to see if the oligomerization can induce hypersensitive response (HR-PCD), a form of programmed cell death (PCD) defense response, in Nicotiana benthamiana. The TIR domain was fused to green fluorescent protein binding protein (GBP) and mCherry fluorescent protein binding protein (ChBP). Through the use of Agrobacterium-mediated transient expression, we co-expressed TIR-GBP and TIR-ChBP with GFP-mCherry fusion in N. benthamiana leaves in order to create a  TIR domain homodimer. Our initial results suggests that artificially induced TIR domain dimerization using this approach fails to induce cell death.

Characterization of a Diterpene Synthase in Grindelia robusta

Student Name: Esha Chawla1
UCD Department: Plant Sciences
UCD Mentor: Dr. Philipp Zerbe

Plants have a diverse set of defense mechanisms in order to protect themselves against external stressors. Among these defense mechanisms is a class of 20-carbon molecules called diterpenoids. Diterpenoids are key in the structure and function of plants, as they protect the plants against pests/insects, excessive UV radiation, and drought [3].  In order to better understand how diterpene synthase, the enzyme that aids in the production of diterpenoids, function in plantae, site-directed mutagenesis was performed near the active site of the diterpene synthase, diTPS. In this study, E. coli bacteria was used as a transient expression system via transformation of E. coli with mutants of GrTPS2 by inducing mutations into GrTPS2 observing the difference in metabolite formation via gas chromatography mass spectrometry analysis (GC-MS). Using this analysis, we can identify which amino acids are key in the structure and function of diterpene synthases. Using this information, future experiments can be conducted to observe the effects of other amino acid substitutions at other positions in the

Ultra-conservation identifies exons missed by traditional annotation

Student Name: Gabrielle Berman
UCD Department: UC Davis Genome Center
UCD Mentor: Dr. Ian Korf

Motivation: Multi-species genomic alignments show that there are numerous regions that retain 100% nucleotide identity even in organisms that last shared a common ancestor more than 300 million years ago. Such extreme conservation is indicative of molecular function, although it is not always clear what that function is. Where such conservation overlaps exons found in another organism, it can be inferred that the underlying sequence is from a gene. In this paper, we sought to discover highly conserved exons missed by previous annotation efforts.
Results: Using a bioinformatics approach, non-human exons were aligned from the Ensembl database to the human genome and proteome. Exons that are highly conserved in the genome but not found in the proteome represent missed exons. Some of the missed exons turned out to be errors in annotation. The most common case was exons that Ensembl failed to annotate even though they were found in other databases. Another source of error were small RNAs that were not masked in the non-human genomes. We also found several cases where the human genome is missing an exon that other related genomes clearly contain. Since these exons are identical to the human genome, it’s highly likely they continue to function. The reason they are unannotated is probably because the expression profile is highly limited, for example, from an early developmental stage.

Modeling Theoretical Plant Systems with Asynchronous Populations

Student Name: Garrett Gould
UCD Department: Plant Sciences
UCD Mentor: Dr. Mohsen B. Mesgaran

Modeling reproductive asynchrony and developing a framework to calculate seed production is very crucial need not only in the further understanding of how asynchronous species reproduce but also in the controlling of asynchronous invasive species. To model reproductive asynchrony, four probability distributions are employed, representing the population density of the male or female of the species that are reproductively active at a given time. This framework models the percent of the population mating, the total amount of seed production, and how these quantities are affected by varying the distribution parameters. By knowing how many seeds will be germinated, given reproductive timing data, further methods of controlling invasive species can be proposed, leading to agroeconomic growth.

Genome-wide Association Study of Plant Development in Arabidopsis thaliana

Student Name: Gaurav Ghosal
UCD Mentor: Dr. Daniel Runcie

Plant development and its influences are relevant to a variety of applications and research questions. However, plant development is multi-faceted as it involves genetics, the environment, and different stages of development. A genome-wide association study (GWAS) was conducted to identify quantitative trait loci (QTL) which control the vegetative development in Arabidopsis thaliana, using a panel of 70 worldwide accessions. Previously collected plant images were fed through a computer vision pipeline in order to extract measurements which, along with existing reproductive trait measurements, were used for association. Established single locus association models were used in addition to a newer, multi – loci model. Afterwards, the relationships between the QTL and traits observed during both developmental stages were analyzed.  Gene annotation was conducted to further investigate the QTL identified in this study and validate the results. This study identified significant QTL for traits in both the developmental stages analyzed. Gene annotation revealed that two QTL associated with vegetative development had overlap with previously found development related genes. Ultimately, the path model indicated a significant indirect effect between a region controlling vegetative development and traits measured during the reproductive stage.

The effects of environment stress on the genetic diversity of Agrobacterium tumefaciens

Student Name: Hanlin Sun
UCD Mentor: Dr. Dan Kluepfel

For over a hundred years, Agrobacterium tumefaciens has been known to cause crown gall disease in trees around the world. Because the Agrobacteria operate under similar conditions, it would logically follow that it would have a narrow genetic diversity. However, Bosmans et al. (2015) has proven through a variety of tests that Agrobacterium contains a uniquely large genetic pool. To gain a glimpse into the factors that create this uncommon genetic and phenotypic variation, the 186r strain of Agrobacterium, a real-world infectious strain that exists today, was placed under a variety of environmental stresses. The Agrobacteria were cultured for two weeks in a soil environment, a water environment, a minimum medium (M9), and a rich medium (TSB). These stress environments would illustrate if the different biotic and abiotic factors of varying habitats cause genetic changes in the Agrobacteria. In addition to these stress environments, strains of 186r Agrobacterium that were stressed for two years in fallow and orchard soil environments were also included for DNA analysis. This would illustrate how long it takes for the environment to instill DNA changes. To show if environmental factors played a role in creating genetically different Agrobacteria, the stressed Agrobacteria underwent a variety of single gene and complete genome scans that show DNA changes at the base level. Not only would this offer explanations for the creation and plasticity of Agrobacterium’s genetic diversity, this would also contribute towards finding a potential solution for Agrobacterium.

Effect of Ibuprofen on Murine 26S Brain-Specific Proteasomes in Cytosol

Student Name: Henry Huynh
UCD Department: Neurobiology, Physiology, and Behavior, & Physiology and Membrane Biology
UCD Mentor: Dr. Aldrin Gomes & Dr. Celena Lozano

Ibuprofen, a NSAID (non-steroidal anti-inflammatory drug), is a popular over-the-counter analgesic and antipyretic, which functions by non-specifically inhibiting the COX-1 (cyclooxygenase-1) and COX-2 (cyclooxygenase-2) pathways. The temporary inhibition of the COX (cyclooxygenase) pathways by ibuprofen reduces the formation of prostaglandins, which are lipids necessary for inflammatory processes. It has been hypothesized that ibuprofen and other NSAIDs could endanger the functionality of the proteasome in the brain. The UPS (ubiquitin-proteasome system) is essential for intracellular protein degradation in mammalian cells. It has been further speculated that ibuprofen could target pathways crucial for proteasome function and cause dysfunction. Misfolded protein aggregates and oxidative stress due to proteasomal dysfunction are intimately linked to aging, neurodegenerative diseases, and cancer.  To determine if ibuprofen induces proteasomal dysfunction in brain tissue, we performed semi-quantitative western blotting and proteasome activity assay analysis on mice brain homogenates to determine the effect of ibuprofen on brain homogenates. The β5 and β2 subunit proteasome activity was found to be statistically significantly reduced in brain tissue derived from ibuprofen treated mice. The heat shock protein 70 (HSP70), which sometimes is upregulated during stress, and β1 are both potential correlates with proteasome function and their activity was not increased in ibuprofen treated mice.

A Study on the Effects of Ibuprofen on Murine 26S Liver-Specific Proteasomal Dysfunction

Student Name: Joon Y. Park
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Aldrin Gomes

Ibuprofen is one of the most common type of nonsteroidal anti-inflammatory drugs used for moderating pain in human body by inhibiting the production of both COX-1 and COX-2. More than 30 million Americans use them to soothe discomforts and pains everyday, according to American Gastroenterological Association (Griffin, 2015). Ibuprofen is well known to show detrimental effects in damaging cardiac and gastrointestinal cells, but no research has yet conclusively demonstrated the clear effect of ibuprofen on the liver. Thus, the goal of this research is to determine the effect of ibuprofen on proteasomal function in liver. Proteasome serves as a useful indicator, as it is responsible for degrading most of the protein in the eukaryotic cells (Budenholzer et al. 2017). Through western blotting technique, the relative amount of proteasome and antioxidant protein was measured. Through proteasome assay, the proteasomal activity was measured. These two approaches showed that ibuprofen caused proteasomal dysfunction in the specific proteins and beta-subunits: PSMA6, β1, and β2. Overall, the ibuprofen was part of the causing factor of 26S liver-specific proteasomal dysfunction. These results suggest that people with liver problems or people at age 45 or older should be cautious about the use and dosage of ibuprofen. This research also suggests that investigation of other NSAIDs, such as aspirin, using similar approaches should be carried out to accurately evaluate hazards on the human body.

Expanding the Database of Single Point Mutations of the β-glucosidase B Protein Using Kinetic and Thermal Analysis

Student Name: Lauren Fu
UCD Mentor: Dr. Justin Siegel

The well-documented crystal structure of Beta-glucosidase B, or the BglB protein, leads to its common usage in studies done on site directed mutagenesis. In terms of this project, the predictions of a computational enzyme design program is assessed using the experimental data BglB mutations. Using a combination of dry and wet lab techniques, three mutations are studied in how changes in enzyme structure affect amino acid interaction. FoldIt is used to predict the changes brought to the wild type BglB protein by the each of the mutations, and the kinetics and thermodynamics are determined through assays on proteins containing the mutant DNA. The data points are evaluated against the projections of the algorithm in order to examine its accuracy. The data reveals that the H206R mutation is more stable than the wild type, as it denatures at a higher temperature than the wild type. [insert kinetic data analysis here]. As the database expands, the predictions of FoldIt can be improved through machine learning.

Effects of Tactile Enrichment in Sensory Cortex and Behavior of Early Blind and Sighted in Monodelphis Domestica

Student Name: Luyan Zhang
UCD Department: Center for Neuroscience
UCD Mentor: Dr. Leah Krubitzer

Compensatory cross-modal plasticity is the ability for the brain to alter neural connectivity and functionality, and it is highly experience-dependent. Rearing animals in enhanced environments has been shown to ameliorate deficits caused by neurological diseases and promote the development of the visual system. The goal of this study was to determine the effects of tactile enrichment on the behavior and cortical area size of early-blinded and sighted short-tail opossums (Monodelphis Domestica), seeking to find the extent enhanced environment can affect cross-modal plasticity. The opossums performed two naturalistic behavior tasks: the variable ladder task and skilled forelimb reach task. While the variable ladder task tests the ability of tactile discrimination, the skilled reach task tests the motor ability and sense of smell. The brains were then extracted from enucleated and sighted animals from enriched-reared animals and standard reared cages. The experiment then involved histological tests to determine the size of primary visual and somatosensory cortices. This study showed that opossums reared in tactile enriched environment made significantly less error in the variable ladder task and skilled reach task, and performed generally better in the behavioral tasks. While the area of primary somatosensory and visual cortices stayed relatively the same for both enriched-reared animals and the standard-reared animals, the density of neurons, their firing rates, or receptive field sizes may have changed. The findings of this research may promote better rehabilitation methods for blind patients.

A Semiquantitative Fluorescent Method to Assess Genome Editing Efficiency and Knock-in Rates using CRISPR/Cas9 and a single stranded DNA oligo

Student Name: Nathaniel Kim
UCD Department: Mouse Biology Program
UCD Mentor: Dr. Joshua Wood

One method for generating animal models for precision medicine applications is to insert pieces of DNA that code for changes in that animal to more closely mimic disease conditions. At the UC Davis Mouse Biology Program, a method has been developed for generating a single stranded DNA (ssODN) construct that can be used in combination with CRISPR/Cas9 to generate mutant animals with DNA insertions. We have employed this method to generate a Cy3 (red) fluorescently labeled ssODN that encodes green fluorescent protein (GFP) and engrafts via small homology arms downstream of exon 1 in the embryonic development gene, Platr 14. To test the durability of the ssODN, we evaluated its structural integrity under repeated freeze/thaw cycles and/or electroporation conditions. By evaluating the resilience of the single stranded DNA to withstand common procedures used in generating the mutant animals, we provide evidence that this method for generating ssODNs is a reliable platform for generating mutant animal models by DNA insertion. Further, by testing the Cy3 fluorescently labeled ssODN that encodes GFP and integrates into a gene that is active on embryo days 4-5, we will be able to measure the relative efficiency of ssODN entry into mouse zygotes and its ability to integrate into the genome by culturing the embryos ex vivo. This innovative tool allows us to rapidly assess the relative efficiency of two electroporation methods in comparison to traditional pronuclear microinjection. Our results showed that only zygotes which underwent BioRad electroporation glowed green on day 5 which shows that integration of the ssODN was successful into the Platr 14 gene. This ultimately shows that the mean fluorescent intensity method is a reliable way to rapidly determine successful integration of DNA using CRISPR/Cas9 and targeted integration of a relatively large ssODN (1692bp) into the mouse genome using CRISPR-EZ.

Relations of Organs and Water Potential to Bent Neck in Roses

Student Name: Nitya Kotha
UCD Department: Plant Sciences
UCD Mentor: Dr. Kenneth Shackel

Bent neck is a common phenomenon seen in roses when the top part of the rose bends over, contributing to a significant monetary loss in the horticulture industry. Bent neck is caused by loss of pressure in the peduncle, but it is unknown what leads to this change in water pressure. In this project, four experiments were conducted, in which water potential was determined by using pressure chambers. The first experiment was conducted to test which organ has the greatest effect on the loss and gain of turgor pressure. The water absorption, content, and loss of each rose was calculated and the water potential of the peduncle, stem, and leaf of each rose was measured. The bud was seen to contribute greatly to water loss in the roses. A second experiment was conducted to test if the presence of the rose bud affects equilibration of the flower by having two flowers without buds and three with buds and proceeding to measure the water potentials of the peduncle, leaf, and stem after equilibrating the roses for three hours. It was found that on average the leaves had great stress values than the peduncle, which was greater than the stem regardless of the presence of the bud. The third experiment tested the presence of the rose bud in relation to the presence of water, having four conditions in total. Using three roses per condition and letting them sit overnight, water potential measurements were taken of the peduncle and leaves of each rose, showing that the bud only plays a role in water stress when a water source is present. The final experiment tested if the presence of leaves affected water loss based on the findings of experiment 1. Using the cultivar Frederic Mistral, it was found that the presence of leaves contributes to more water loss. Further research must be conducted to test the applications of the research.

Application of the hydrotime model to germination in invasive weeds and emergence from various burial depths

Student Name: Quin Wai Wong
UCD Department: Plant Sciences
UCD Mentor: Dr. Mesgaran M. Matzrafi

The hydrotime model is a common system for predicting seed germination in response to water availability. Developed after the thermal time model, its functionality lies in accounting for the environmental conditions of seeds in a statistical fashion. The hydrotime method focuses on water potential: the ability of a plant to take in water. By comparing water potential to germination rates, more data can be gained by applying the hydrotime model to various stressors. The seeds are germinated at nearly optimal temperature with varying water potentials through incubation. Since Palmer amaranth has high genetic variability in its population, and hence, varied times for seed germination, it is important to understand whether the hydrotime system can be applied. Through analysis on hydrotime for Palmer amaranth, more questions arise as to whether either the hydrotime or thermal time model have applications to other environmental conditions. Analysis of emergence in terms of seed burial depths with similar modeling techniques is approached. The hydrotime model best matched the emergence studies on seed depth. However, use of the logistic distribution and a separate emergence equation were required. From this, the amount of parameters required to express emergence are decreased for seed depth by examining trends in the data. As a result, this study benefits growers in managing invasive weeds while providing insight on the implications of burial depth modeling.

Observing the Influence of the Presence of Rose Leaves, Flowers, and Peduncles on Water StressThrough Water Potential

Student Name: Rhea Cho
UCD Department: Plant Sciences
UCD Mentor: Dr. Kenneth Shackel

The floriculture industry experiences losses due to premature wilting by flowers during transport every year. Water potential has been recently proposed as an effective and accurate indicator of plant stress levels in a variety of crops, including flowers. The primary objective of this study was to observe the water relations of roses (Rosa) with respect to the presence of certain rose organs in detail through four experiments. ANOVA tests were conducted to detect overall significance of data, and the Tukey Test was used to make direct comparisons between treatment groups. In the first experiment, ten roses of two different cultivars (‘La Perla’ and ‘Ingrid Bergman’) were cut in five ways to create different organ combinations. A pressure chamber measured the water potential values of the stems, leaves, and peduncles of these roses after four days. It was concluded that organ combinations with flowers exhibited more negative water potential values under conditions of water stress. The second experiment involved observing the influence of the presence of a flower on water equilibration of roses of one cultivar. No significant difference in water potential values was seen between stem to stem, peduncle to peduncle, and leaf to leaf comparisons between roses with flowers and roses excluding flowers. The third experiment investigated the differences of water potential values of the stems, leaves, and peduncles of twelve roses subjected under four different treatments: with flower and with water, with flower and without water, without flower and with water, and without flower and without water. A statistically significant difference was found between with water and without water treatments, but not between with flower and without flower treatments. The fourth experiment was essentially a repeat of the first experiment with a greater focus on observing the influence of the presence of leaves on water potential values. A statistically significant difference was found in water gain in rose, uptake, and water potential values. These findings indicate that the presence of flowers and leaves negatively impacts rose water potential values in post-harvest conditions. Practical applications for this research involve further exploring which pre-harvest growth conditions for roses reveal more negative water potential values for assessing the effectiveness of specific water or soil treatments. 

Markers of Mitochondrial Dysfunction in FMR1 Premutation Carriers

Student Name: Roshni Varma
UCD Department: MIND Institute
UCD Mentor: Dr. Cecilia Giulivi

A premutation in the fragile X mental retardation-1 (FMR1) gene is characterized by 55 to 200 repeats of the trinucleotide CGG in the 5’-untranslated region of the gene. Some premutation carriers will develop a late-onset neurodegenerative disease known as fragile X-associated tremor/ataxia syndrome (FXTAS). The mechanisms that induce FXTAS in premutation carriers remain elusive, but mounting evidence points to age-dependent accumulation of mitochondrial damage, which is exacerbated by the CGG expansion. Past research reported decreased citrate synthase activity and mitochondrial DNA (mtDNA) copy numbers, coupled with increased mtDNA damage in premutation carriers with and without FXTAS. However, the link between citrate synthase and the mtDNA copy number (both usually used as surrogate markers of mitochondrial mass) has not been investigated in detail in premutation carriers with and without FXTAS. This is relevant in the context that a higher mtDNA copy number may be associated with oxidative stress, whereas a lower copy number may be associated with issues of mtDNA maintenance and replication. To this end, this study evaluates citrate synthase activity and mtDNA copy numbers per cell in skin fibroblasts from nine premutation carriers, six of which were diagnosed with FXTAS. The majority of carriers (5 of 9) showed lower mtDNA copy numbers than the 95% confidence interval (CI), whereas only one of nine showed lower citrate synthase activity. While more research is needed within a larger dataset, these results indicate that lower than control values of mtDNA are present in carriers.

Enhancing Determinants of Tyrosine Sulfation Site Predictions

Student Name: Samarth Kamle
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Grace Rosenquist

The post-translational modification of tyrosine sulfation is thought to be recognized when the sulfating enzyme tyrosylprotein sulfotransferase exhibits specific properties. However, with the recent discoveries of new sulfated sites, recognition features to identify tyrosine sulfation require modifications. Thorough analysis of amino acid sequences using the Position-Specific-Scoring-Matrix (PSSM) emphasizes that the consensus features do not possess updated information essential to predict tyrosine sulfation. The amino acid residues surrounding the tyrosine site affect the condition of the tyrosine as sulfates tyrosines are typically found near acidic or mutagenesis sites. New sulfated tyrosines with low PSSM scores indicate alternate factors contributing to the status of the tyrosine site as the cutoff score has decreased. This view implies that current consensus features to recognize TPST substrate may not be essential as reevaluation of the factors contributing to prediction of tyrosine sulfation have revealed lower sensitivity values for the dataset of known sulfated tyrosine sites (p114n551).

Optimization of RAFT-forming Drug Formulations

Student Name: Sarah Suriano
UCD Department: Biological and Agricultural Engineering
UCD Mentor: Dr. Gail Bornhorst

Gastroesophageal reflux disease, or acid reflux, causes 25-100 million people to experience heartburn in the US alone. Treatments available include antacids, raft-forming drugs, and combinations of the two, which consist of sodium alginate, calcium carbonate, and sodium bicarbonate, which react together to form a raft that floats to the top of the stomach contents to prevent acid from entering the esophagus. The purpose of this study is to investigate the effects of the composition of the formulation on pH and raft formation. Each of the different formulations was tested in vitro in HCl solutions of pH 1 and pH 2. The mass of the raft and the pH of leftover solution were measured and the effects of the ratios and composition of the formulations on the pH and raft were analyzed. The results suggest that 4.5 g of sodium alginate is the optimum level because it always forms a gel.  

Optimization of AtCBF1 ectopic expression in harvested tomato fruit using a chemically-inducible transgenic system

Student Name: Sarah Wornow
UCD Department: Plant Sciences
UCD Mentor: Dr. Diane Beckles

        Tropical and subtropical fruit, such as the tomato, experience postharvest chilling injury (PCI) when stored at low temperatures, accelerating deterioration and reducing shelf-life, resulting in losses for the agricultural industry and consumers. CBF genes (CBF1-3) code for transcription factors which regulate the expression of cold responsive genes that confer tolerance in the model plant Arabidopsis thaliana. Overexpressing a CBF gene from this plant (AtCBF1) in tomato fruit during postharvest refrigeration may increase PCI tolerance and better preserve quality. The goal of this project was to optimize the application of the glucocorticoid dexamethasone (Dex) in transgenic Micro-Tom tomato fruit containing a construct to chemically-induce AtCBF1 expression during periods of refrigeration. Transgenic tomato fruit were harvested, soaked in a Dex solution, and refrigerated for specific times. RNA was extracted to quantify the relative expression of AtCBF1. A physiological characterization found control seeds (no AtCBF1) had a higher germination percentage than the transgenic seeds at Day 8. The optimal concentration of Dex to induce gene expression was found to be fifty micromolar and the optimal soaking time to be one hour. In conclusion, the Dex system successfully induced AtCBF1 expression in harvested fruit, a novel finding.

Refinement of quantification methods for Sporosarcina pasteurii

Student Name: Sarah Yun

Aims: Sporosarcina pasteurii is a bacterium used to precipitate calcite in sands, creating a sandstone-like material. As it has been difficult to attain a viable count of S. pasteurii comparable to that of direct counting methods, this project aimed to refine quantification methods for S. pasteurii.
Methods and Results: S. pasteurii cultures were serially dilute and viably plated on various medium to compare the ability of the medium to facilitate efficient cell recovery. It was found that agar media containing nutrients mixed for longer periods of time facilitated the best recovery. Additionally, the relationship between S. pasteurii optical density and total direct count by acridine epifluorescence microscopy was compared across spectrophotometers to find significant discrepancies.
Conclusions: Agar media containing nutrients mixed for longer periods of time and low optical density inoculums can be used to meet expected recovery. Further studies should aim to verify these findings. The discrepancies across spectrophotometers suggest researchers must not solely rely on data from outside spectrophotometers and must instead develop a correlation for each spectrophotometer.
Significance and Impact of Study: This project enables the accurate physiological study of soil bacteria and improvement in their utilization by demonstrating proper quantification.

The ROG2 Protein Interacts with Arabidopsis TRAPPIII Tethering Complexes and is Involved in SYP61 Trafficking

Student Name: Sean Fu
UCD Department: Plant Sciences
UCD Mentor: Dr. Georgia Drakakaki

It is known that cell wall components are trafficked by vesicles that fuse to plasma membranes mediated by SNAREs and tethering complexes. Highly conserved tethering complexes such as TRAPP complexes are known to guide vesicles and fuse them to the plasma membrane. In Planta, TRAPPIII complexes have been identified. Recently, research done has shown a new protein called ROG2 within the complex. Here, the initial characterization and interaction between ROG2 and other well-known TRAPP complex proteins is investigated. One component of this characterization was done by analyzing the relationship between ROG2 and known TRAPPIII complex components such as TRS33 and BET3. The presence of ROG2 in TRAPP complexes was demonstrated by creating biomolecular fluorescence complementation (BiFC) using a 2-in-1 plasmid containing ROG2 along with a potential TRAPPIII binding partner (such as TRS33 and BET3). The interaction between ROG2 and the TRAPPIII proteins was analyzed using confocal microscopy. The images revealed that ROG2 does interact with (insert which one it interacted with), suggesting that ROG2 is part of a new TRAPPIII-like complex. Two mutant Arabidopsis lines, rog2-1 and rog2-2, had their rog2 genes knocked out to better understand the role of ROG2 in cell wall component trafficking. The mutations resulted in the disruption of normal trafficking conducted by SYP61 vesicles. Work was primarily done with rog2-2 and the two recovery Arabidopsis lines CFP-SyP61xROG2-2 Dipped w/UB-YFP-ROG2 and CFP-SyP61xROG2-2 Dipped w/Rog2-YFP-ROG2. The rescues of these mutants verified the lack of the ROG2 protein was responsible for the cell dysfunction.

Magnetophoresis of Iron Oxide Spherical Nanoparticles and Anisotropic Nanorods

Student Name: Sean Wu
UCD Department: Chemistry
UCD Mentor: Dr. Ting Guo

Two competing morphologies of iron oxide (Fe3O4) nanoparticles, spherical and rod shaped, were synthesized via established solvothermal and three-step (hydrolysis, reduction, and ligand exchange) reaction methods, respectively. Single crystal spherical particles averaging 30.8 nm in diameter and nanorods averaging 40.3 nm in length and 9.9 nm in diameter were synthesized and dispersed into an aqueous biofluid model solution, under a magnetic field gradient. The field strength of a permanent magnet was measured and found to fall off quickly past 2 cm. Movement experiments were conducted between 0-1 cm away from the magnet, with a field of 3180 Gauss at the surface and down to 750 Gauss 1 cm away from the magnet. It was anticipated that despite similar size and component materials, the anisotropic characteristic of nanorods would attribute to the faster induced magnetization movement in media relative to its spherical counterpart, suggesting a necessary shift of focus towards anisotropic magnetic nanostructures. Further work is needed to improve crystallinity and size matching in the rods used in this work to realize the improvement in magnetic response compared with equivalent spheres. The preliminary results with 40.3 nm 9.9 nm rods which are smaller in material volume and less crystalline than the spheres point towards the anticipated results.

Comparing the Breathing Structures of Cactophillic Drosophila Larvae and Pupae to Other Species

Student Name: Serena Wong
UCD Mentor: Dr. Susan Lott

Evolutionary adaptations in many species of organisms prove to be protective mechanisms against predators and prey alike. Like many other species, cactophillic Drosophila have developed adaptations, such as long epithelial breathing tubes, that enable them to survive in harsh desert climates. However, the cause for developmental differences in Drosophila breathing tubes is not clearly understood by the scientific community. This research project studied 4 different species of Drosophila: three cactophillic species and one non-cactophillic species as a control. Specimen were first imaged using a stereoscopic microscope and linear measurements were taken of different pupal body segments. Using specialized computer software, the measurements were analyzed and comparisons were conducted between anterior and posterior spiracles against full body lengths. The results show that there is little to no difference between the length of the two anterior spiracles on each pupa, while there is a difference between the two posterior spiracles of each pupa in either cactophillic or non-cactophillic Drosophila. Ratios between anterior or posterior arm segment lengths and whole body lengths were mostly uniform within species, but vary between species.

The function of the homologous recombination Dmc1 protein during zebrafish meiosis

Student Name: Tina Li
UCD Department: Molecular and Cellular Biology
UCD Mentor: Dr. Sean Burgess

Meiotic recombination is necessary for generating genetically unique gametes, homologous chromosome pairing, and segregation. Recombination initiates with programmed DNA double strand breaks (DSBs). These DSBs are resected to reveal 3’ single strand DNA, which are bound by recombinases Dmc1 and Rad51. These recombinases mediate DNA homology search and strand invasion, which results in chromosome pairing. However, when recombination defects occur, it can lead to chromosomal abnormalities or miscarriages.
To test the function of the dmc1 gene, a dmc1 knockout strain in zebrafish was created. Then, the dmc1+/- zebrafish were crossed together. The offspring from the heterozygous dmc1 zebrafish  were genotyped and separated into three categories: dmc1+/+, dmc1+/- or dmc1-/-. It is discovered that in the dmc1-/- group, the majority of the progeny was female. Another discovery was made after crossing the female offspring with wild type males. The survival rate, which was 98.120%, was the highest after crossing the dmc1+/- and WT fish. The survival rate of the two other crosses were 93.671% (dmc1+/+ and WT cross) and 93.130% (dmc1-/- and WT

High-amylose resistant starch mutant wheat (Triticum turgidum) shows a change in germination pattern under cold stress

Student Name: Tomo Yoshino
UCD Department: Plant Sciences
UCD Mentor: Dr. Diane Beckles

High-amylose wheat is highly desired because it resists digestion to sugars and thus acts as fiber with all the associated health benefits. A mutant wheat (abAB) with twice the amylose content of normal wildtype (WT) wheat was engineered by disabling a starch branching enzyme. The hypothesis is: wheat germination will not adapt to high resistant starch as it would not be able to digest enough starch for use. The germination of these variants was compared over 12 days at room temperature ® and in the cold (C) at 15°C. Low temperatures slowed germination at 4 days. By day 12, accelerated growth in WT-C made coleoptile and radicle length, water uptake, and biomass distribution the same as in the non-stressed WT-R seeds. In contrast, the mutant showed greater defects in the cold; it only adapted its coleoptile and radicle length at day 12. There was surprisingly, a higher amount of both sugars and starch in cold-treated abAB seeds at day 8, suggesting altered metabolism of these carbohydrates in the mutant. Overall, wheat germination adapted to high levels of resistant starch and low temperatures by day 12, since there were few differences in physiological and biochemical parameters assayed compared to the control.

Ablation of somatostatin-secreting Delta cells and its effect on the maturation of Beta cells

Student Name: Vivek Annadata
UCD Mentor: Dr. Mark O. Huising & Jessica L Huang

The islets of Langerhans in the pancreas consist of alpha, beta, and delta cells. The focus of this research is to understand the paracrine interaction between the pancreatic delta and beta cells and how their impaired interaction contributes to diabetes development. Specifically, our goal is to determine the effect of somatostatin secreted by delta cells on the maturation of beta cells in the Islets of Langerhans. We studied this by comparing the levels of Urocortin 3 (Ucn3) secreted by mature beta cells in normal mice with levels of Ucn3 in mice whose delta cells have been ablated. Compared to the wild type normal mice, we found negligible levels of somatostatin but the equivalent levels of Ucn3 in the specimens with delta cells ablated. These results offer an explanation that either somatostatin has no effect on the secretion of Ucn3 and thereby on the maturation of beta cells or for somatostatin to exhibit an effect, the pancreas has to be acquired after waiting for a longer duration than 6 days, as was done in this study. 

Effects of Multi-walled Carbon Nanotubes on the Lung Cells of Mice and Rats

Student Name: Younkyung Oh
UCD Department: Center for Health and the Environment
UCD Mentor: Dr. Kent Pinkerton

As advancements in nanotechnology are drastically improving consumer products in recent years, the human health effects and environmental safety issues associated with nanomaterials remain widely unknown. The primary objective of this study is to determine whether multi-walled carbon nanotubes (MWCNTs) inhaled into the lungs 1) cause harm to the respiratory system via inflammation, 2) can be cleared from the lungs over time and 3) if not cleared, where in the lungs they can be found. Inhalation methods used for this study include the exposure of mice and rats to three different concentrations (0.06, 0.2, 0.6 mg/m³) of aerosolized MWCNTs in exposure chambers each weekday (Monday-Friday) for approximately one month. Following end of inhalation exposure, B6C3F1/N mice and Harlan Sprague Dawley rats were examined at two different periods of time post exposure (PE), at approximately one and five weeks. Our findings demonstrate a dose-dependent acute inflammatory response that subsides by the 4-5 week PE time. Continued retention of MWCNTs raises concerns regarding possible long-term effects.

2017
 

The corC Gene Influences Bacterial Sensitivity to Two-Peptide Bacteriocin Plantaricin EF

Student Name: Alice Gevorgyan
UCD Department: Food Science and Technology
UCD Mentor: Maria Marco

Plantaricin EF is an antimicrobial peptide produced by Lactobacillus plantarum to eliminate competition for food and resources. The exact mechanisms through which Plantaricin EF (plnEF) causes cell death are not known, but research in this lab (manuscript pending) has suggested that plnEF binds to magnesium channels on a cell’s surface to cause membrane leakage and cell death. The objective of the current study was to provide evidence that corC, encoding a putative magnesium efflux pump, serves as the molecular receptor for plnEF. In order to determine if there was a connection between the corC gene and plnEF susceptibility, two strains of L. plantarum and nine strains of L. pentosus, all of which are very closely related to the plnEF producer L. plantarum strain 8826, were isolated and used for DNA extractions. The corC genes from these isolates were sequenced and their sensitivities to plantaricin were measured using 96-well microtiter plate assays. By combining the data from the sequencing and the sensitivity assay, it was found that sequence divergence of the corC gene was associated with altered sensitivities to plnEF. This research can help clarify the mechanisms through which plnEF acts and fill in some of the immense gaps in knowledge that exist in the understanding of bacteriocins. Eventually, this research can help other researchers turn bacteriocins into a new class of antibiotics and aid in the incorporation of plantaricin into the food industry to provide safer, cleaner food.

Determining conserved motifs that distinguish high-expressed introns from low-expressed introns with a genetic algorithm on the C. elegans genome

Student Name: Allen Mao
UCD Department: Molecular & Cellular Biology
UCD Mentor: Ian Korf

In a wide variety of eukaryotes, some introns elevate mRNA accumulation to increase gene expression. However, the biological mechanism for this process is not well understood and it is unknown why some introns have this effect while others lack it. In this research, the object is to have a better understanding of intron function to gain greater control over gene expression by creating a genetic algorithm that identifies sequence motifs that are more commonly found in introns of high expression than introns of low expression. The approach for creating the genetic algorithm consists of generating random motifs, selecting fit motifs as per an objective and fitness function, and randomly mating and mutating motifs in order to yield high surviving motifs against the objective and fitness function.

The Effect of Mutations in Tropomyosin on Their Susceptibility to Protease-Facilitated Digestion

Student Name: Amy Du
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Aldrin Gomes

Tropomyosin works in conjunction with the troponin complex to regulate muscle contraction and relaxation. It is an alpha-helical protein that extends over the actin’s myosin binding sites. When calcium binds troponin, troponin interacts with tropomyosin in a way that changes tropomyosin structure, allowing myosin to bind to actin. Tropomyosin mutations are known to cause hypertrophic and dilated cardiomyopathy. However, the pathway for this is unknown. Preliminary studies conducted on tropomyosin studied the presence of phosphorylation sites and the up or down regulation of their phosphorylation, suggesting a structural change in the mutants. The presence of phosphorylation sites on tropomyosin were recently found but the functions of these sites are not known. Altered phosphorylation levels are usually important in protein function. To explore if mutations or phosphorylation of tropomyosin affect its conformation digestion of different tropomyosins was carried out. Trypsin digestion of tropomyosin mutants compared to the wild type suggest that the change of a single amino acid sequence affects digestion, which may be related to the structure. Since structure dictates function, a change in shape suggests a change in function. The implications of these results allow us to see the potential mechanism (structural change) for which mutated tropomyosin proteins may cause cardiomyopathy.

The Effects of Varying Concentrations of Different Chemicals on the Growth Rates of Microcystis aeruginosa

Student Name: Andrew Chen
UCD Department: Department of Anatomy, Physiology, and Cell Biology
UCD Mentor: Dr. Swee Teh

Microcystis aeruginosa is a unicellular, colonial cyanobacterium that produces a potent class of hepatotoxins called microcystins (MCs). During harmful algal blooms (HABs), Microcystis populations can drastically increase and release substantial quantities of microcystins into the surrounding waters upon cell death. These toxins can compromise environmental and human health. Therefore, it is important to understand what factors inhibit or stimulate Microcystis growth. In this study, Microcystis cultures were exposed to Roundup (glyphosate), bisphenol A (BPA), Diuron, and lab-created microplastic solution in a 96 well plate. Growth of the cultures were tracked daily by measuring the absorbance of chlorophyll in each well with a spectrophotometer. After three days, treatments that exhibited substantial growth inhibition or stimulation from the control group were scaled up to 250 mL flask exposures. A more concentrated microplastic solution was created for the flask exposure in order to explore the effect of a more concentrated exposure. The results of this study revealed that exposure to Diuron severely inhibited the growth rates of Microcystis. BPA and glyphosate inhibited growth in the well plates, but stimulate growth in the 250 mL flasks. Exposure to the microplastic solution inhibited growth more severely in the flasks than in the 96 well plate.

The Role of Tyrosine Sulfation in Liver Regeneration

Student Name: Archita Bhattacharya
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Grace L. Rosenquist

Tyrosine sulfation, the posttranslational modification of the amino acid tyrosine, has been discovered in countless major proteins in the body, and this study has predicted new sites in the proteins essential to liver regeneration. Liver proteins associated with regeneration were scored in a PSSM calculator. A wide variety of these proteins, including Fibroblast Growth Factor Receptors, Transforming Growth Factorsere found to have high scores, and a collection of predicted sulfation sites were found to also have natural variants. A total of 45 predicted sites were found with scores over the high cutoff of 2.42. In addition, 5 protein sites with high scores were found in clusters. The information on predicted sites could be used to understand regeneration of organs in other organisms and to develop methods aiding the natural process of liver regeneration.

Synthetic and natural auxin plays a major role in solar tracking for sunflowers

Student Name: Breana Lee
UCD Department: Department of Plant Biology
UCD Mentor: Stacey Harmer

Sunflowers undergo heliotropism, the dynamic form of phototropism, which is the directional growth of a plant towards a source of light. The goal of this project is to investigate how sunflowers adjust its growth patterns and respond to changes in the environment to better understand the solar tracking behavior of sunflowers. Hormones, such as auxin, play an important role in plant growth and cell elongation.  In Stacey Harmer’s lab, heliotropism has been studied by testing synthetic and natural types of auxin, identified as 2,4-D and IAA, respectively. In this experiment, IAA, a natural form of auxin, was applied to the east or west sides of sunflower stems at dawn or dusk. To investigate the movement of sunflowers in response to the application of IAA, images were collected measuring the angle between the apex of the plant with respect to the horizon line. Graphical analysis of the angles of curvature showed that when treated with IAA, sunflowers exhibited similar changes from natural heliotropic movement as when 2,4-D was applied. These findings confirm that when applied, auxin, in both its synthetic and natural forms, is responsible for the change in the natural movement of sunflowers and thus proves that auxin has a profound effect on sunflower heliotropism during the day. The knowledge gained by studying heliotropism in sunflowers can be applied to other economically important  plants and may improve the growth and maintenance of these plants for the agricultural industry.

Synthesis of amorphous molybdenum disulfide and tungsten disulfide to be used as a catalyst for carbon dioxide reduction

Student Name: Camille Killeen
UCD Department: Department of Chemistry
UCD Mentor: Jesus Velazquez

Atmospheric carbon dioxide can be used in the carbon dioxide reduction reaction to produce fuels such as 1-propanol, methane, acetate, ethylene glycol and formate from the CO2 reduction using amorphous molybdenum disulfide (MoS2) and tungsten disulfide (WS2) as catalysts. Wet chemical synthesis of these catalysts has been performed and their efficacy for this reaction has been investigated. The MoS2 and the WS2 were characterized to confirm their chemical composition and that they were amorphous. The catalysts were tested as thin films on silicon wafers at a potential of -1.6 V for 15 coulombs (24 hours), testing the efficacy through nuclear magnetic resonance spectroscopy (NMR) to discover the products of the CO2 reduction. While all of the previously mentioned products are useful, the most impressive was 1-propanol. This research may lead to a practical, cost-effective way to produce these fuels from atmospheric CO2 as a renewable source of energy.

Characterization of the effects of inhibiting fatty acid beta-oxidation in Breast Cancer Cells

Student Name: Carl Gibson
UCD Department: Department of Molecular Biosciences, School of Veterinary Medicine
UCD Mentor: Cecilia Giulivi

Differences in the metabolism among a population of breast cancer cells leads to difficulty in fully destroying the tumor. This metabolic heterogeneity was found in glucose-specific pathways and seemed to rely on interdependence among the tumors. This study hypothesized whether cancer cells, within a single tumor, could be separated based on their oxidation of glucose over fatty acids as a fuel source. This was accomplished by utilizing MDA-231 breast adenocarcinoma cells with two different, but directed to the same target (carnitine palmitoyltransferase I or CPT-1), inhibitors of fatty acid beta-oxidation: etomoxir and 4-hydroxy-L-phenylglycine (HPG). Cells were grown for 4-5 weeks on these inhibitors to ensure selection, and then different parameters were evaluated: mitochondrial respiration, cell proliferation, metastasis, pluripotency, and differentiation. The cells that were grown in etomoxir were more metastatic than the control, but had no effect on the other aspects measured in the cells. HPG increased metastatic capacity of the cells, ATP production with octanoate, and cristae density (cytochrome c oxidase over citrate synthase activities), whereas decreased cell differentiation (as judged by ESR1). Neither etomoxir nor HPG had an effect on cell pluripotency. Neither of the treatments affected cell viability or proliferation (evaluated as doubling time). The differences between inhibitors could be due to the presence of the CPT-1b form in breast cancer cells (mainly inhibited by HPG) vs. CPT-1a which is more inhibited by etomoxir. In conclusion, breast cancer cells forced by HPG to grow on glucose oxidation tend to have more metastatic capacity and decreased differentiation, suggesting a shift towards stemness, and developed more active mitochondria, whereas etomoxir-induced glucose oxidation only led to more metastatic capacity.

Comparing and Contrasting the Aspects of PCR and Taqman Assays

Student Name: Chelsea Mai
UCD Department: Mouse Biology Program Center for Comparative Medicine
UCD Mentor: Dr. Joshua Wood

The Mouse Biology Program is a production lab that specializes in producing transgenic and KO mice. To identify whether or not the initial founder mice contain the correct DNA modification, a screening process must take place (Garibyan, Avashia 2014). In retrospect, Sanger sequencing is a method that occasionally can give inconclusive results because the possibility of mosaicism arising due to the inherent multiple mutagenic nature of CRISPR modified genes. Taqman and PCR are more efficient in screening DNA, but due to slightly higher costs of Taqman, PCR vs. Sanger sequencing and the need for accurate data, traditional PCRs and Taqman assays require additional research to discover where they are best applied (Applied Biosystems). This project included the comparison of two pieces of biotechnology: Taqman QPCR and traditional PCR, in order to optimize the efficiency of the screening methods used to look at DNA insertions. Analyzing these screening methods helped provide MBP with the most effective way to produce gene edited mice thus furthering scientific research. The Taqman QPCR and traditional PCR screened a Knock In (KI) of SNPs and of loxP/CRE of the gene mir29a. By analyzing a gel and amplification curve of the screenings of this gene, it can be determined whether a Taqman or PCR is better suited for a particular method.This research contributed to enhancing the efficiency of this production lab. It is essentially a chain reaction: with better accuracy in screening DNA , MBP has more resources to produce gene edited mice at a faster rate thus providing research labs with more mouse models. By creating a library of mouse models, research labs looking for a certain gene function will have easy access to a mouse model with the desired gene function.

Search for a Homozygous Mutation of the LUX Gene in Sunflowers

Student Name: Cheryl Liang

UCD Department: Department of Plant Biology

UCD Mentor: Stacey Harmer

Sunflowers track the sun during the day, moving from east to west from dawn to dusk respectively. However, at night, they move back towards the east in anticipation of the sunrise despite a lack of sunlight. It is known that the circadian clock plays a role in regulating this heliotropism that increase plant fitness. Seeds from a heterozygous mutant plant were planted in an attempt to find a homozygous recessive mutation of the LUX gene that represses day-phased genes in the circadian clock of Arabidopsis thaliana. Using DNA extraction, PCR, and restriction digestion, no plants of the twenty samples were found to be a homozygous recessive mutant. Future research includes performing the experiment again with more samples, and once a mutated plant is found, planting the homozygous mutant plant in a field to observe any phenotypic differences from typical behavior. Understanding how a disrupted LUX gene affects the circadian clock and solar tracking may help future research in improving plant adaptations to increase plant productivity on a molecular level.

Implications of early loss of vision on tactile discrimination and motor capabilities in Monodelphis domestica

Student Name: Chris Iyer

UCD Department: Center for Neuroscience

UCD Mentor: Leah Krubitzer

Loss of vision at an early developmental stage results in profound anatomical and functional alterations in the brain. These neural changes result in cross-modal cortical plasticity, the process by which the deprived cortex is co-opted or taken over by the spared sensory systems. In this study, we investigated the extent to which the neocortex of the short-tailed opossum (Monodelphis domestica) can compensate for the lack of visual input by augmenting somatosensory and motor systems. We examined behavioral correlates of this plasticity by comparing the gross and fine motor capabilities of enucleated and control opossums in a skilled reaching task. Furthermore, we explored the impact of housing conditions on the extent to which visually-deprived brains can adapt and compensate. Our results indicated that the enucleated opossums significantly outperformed the control opossums in both the light and dark conditions, and that tactilely-enriched opossums could potentially demonstrate superior motor control, likely due to enhanced tactile and olfactory capabilities. These results imply that cross-modal reorganization can not only compensate for blindness but allow the animal to outperform normal animals in some sensory-mediated behavioral tasks, offering a foothold for further research into the exact mechanisms involved in plasticity, the brain’s full capacity for remodeling, and potentially improved therapies for sensory-impaired patients.

Physical and Chemical Property Changes during In-vitro Gastric Digestion of Boiled and Fried White Potatoes

Student Name: Daniel Kim
UCD Department: Department of Biological and Agricultural Engineering
UCD Mentor: Gail Bornhorst

As the physical properties of foods are linked to the processes and the functionalities of the digested materials, a demand for the knowledge of how food behaves during gastric digestion from the public has increased. The main objective of this study was to compare two contrasting cooking methods, frying and boiling, that were utilized in order to investigate the differences between the physical property of the boiled and fried white potatoes. The data collected will be part of a broader system that classifies food based on digestive properties: The Food Breakdown Classification System. This classification system aims to predict a food’s digestive behaviour based on its physical properties measured after in vitro digestion. The physical property that was measured from the white potatoes in this experiment was hardness. A Texture Analyzer was used to measure the hardness. The pH and the brix of the digestive fluids were also accounted for. Potato cubes were digested in simulated saliva and gastric juice for six time points of varying length. It was hypothesized that the potato cubes digested for longer time points were to have lower hardness values overall. Also, the pH and the brix of the digestive fluids were expected to increase as the digestion time increased. At the end of the investigation, it was concluded that the fried potatoes had an overall lower hardness than the boiled potatoes. LIkewise, the pH and the brix of the digestive fluids of both boiled and fried white potatoes increased as the digestion time increased.

The relationship between enzyme structure, thermostability, catalytic efficiency of threepoint-mutations in theβ-glucosidase B enzyme

Student Name: Darren Wang
UCD Department: Department of Chemistry
UCD Mentor: Justin B. Siegel

The ability to accurately view a protein in 3 dimensions is very new. Although the images are accurate, the data the models give are faulty due to lack of background data. With a massive database, scientists can compile an algorithm that will greatly help future enzyme engineers. In the future, they no longer need to test every mutant they find interesting in a wet lab, scientists will only need a laptop and a few minutes instead of a weeklong process. So far, the Siegel lab has collected data for 129 mutants of beta-glucosidase enzyme, also known as BglB. For this research, 6 original enzymes are designed using the current computational model, also known as the Rosetta Model, to mutate BglB. Other mutants of the BglB enzyme were grown simultaneously. Lastly, a positive control was used as a benchmark. After the cells were induced with the desired DNA, assays were executed to calculate the thermostability and the Michaelis-Menten constants (kcat, KM, and kcat/KM). Although the research has no short-term results, the small amount of data we can provide is still valuable for the development of a better predictive algorithm. As seen from the data we collected, we can see a general trend of lowering of catalytical efficiencies and a rise in thermostability for the three specific mutants.

The effect of somatostatin on mRNA expression in mouse pancreatic beta cells

Student Name: Eleanor Goh
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Mark O. Huising

Healthy mature beta cells secrete the peptide hormone Urocortin3 (Ucn3) to regulate insulin secretion through a negative feedback loop involving somatostatin, which suppresses insulin secretion. In the beta cells of diabetic mice and humans, Ucn3 levels are significantly lower than those in healthy individuals. The resulting low levels of somatostatin lead to sustained insulin secretion, where insulin production rate lags significantly behind release rate. The stress induced by sustained secretion may then lead to beta cell dysfunction. Adding somatostatin may induce beta cell rest, restoring beta cell function. This study shows the potential of somatostatin in amplifying the beta cell’s acute insulin response, suggesting a key role of somatostatin in the regulation of beta cell maturation markers under high glucose conditions. Finding new methods to provide beta cell rest could lead to new treatments in which beta cell dysfunction can be reversed.

The Emergent Nature of Alignment Effects in Spatial Learning and Memory

Student Name: Elliot Clark
UCD Department: Center for Neuroscience and Department of Psychology
UCD Mentor: Dr. Arne D. Ekstrom

Alignment effects were found to have an emergent nature. Individual differences in the data across participants of two spatial memory experiments were analyzed to find an accurate period in which alignment effects begin to occur. In Experiment 1, participants navigated to goals in a large-scale, virtual environment interspersed with a pointing task to assess spatial memory. The procedure for Experiment 2 was identical to that of Experiment 1, but added an additional task in which participants learn the environment to criteria with a map before exploring the environment. In both experiments, there was a significant difference in performance levels on the pointing task between aligned trials and misaligned trials, but only after repeated exposure to the environment. Though alignment effects are well documented in the literature, little is known about brief period when they manifest. The results confirm that alignment effects are not expected initially, after brief experience, but rather that they arise after learning.

Analysis of TAD calling Algorithms for Hi-C Data Analysis

Student Name: Eric Tang
UCD Department: Biochemistry and Molecular Medicine
UCD Mentor: Fereydoun Hormozdiari

Hi-C, a form of chromosome conformation capture (3C), is a method that is used to create an accurate 3D model of the genome using methods that reveal chromatin structures. However, Hi-C data alone is not enough to produce a stand alone model. Processing steps involving various algorithms are necessary in order to use the read pairs within the Hi-C data to generate the most accurate images of chromatin interactions and topographically associating domains (TAD). The specifications of each of these algorithms, and the disparate results they produce as a result thus require further research. Research within the Hormozdiari lab used statistical and qualitative analyses to look at the different TAD’s produced by the algorithms Arrowhead, Armatus, HiCseg, TADtree, TADbit, InsulationScore, and DomainCaller in order to assess which of them was the most accurate and effective. Although the results of the research revealed the differences in the size, shape, and proximity of the TADs that were found by the algorithms, concrete conclusions regarding the accuracy of the algorithms could not be made

The Q324H Variant of the DNA Repair Glycosylase MUTYH May Predispose CRC

Student Name: Garrett Ma
UCD Department: Department of Chemistry
UCD Mentor: Dr. Sheila David

MutY is a bacterial DNA repair glycosylase that cleaves a mismatched adenine across the oxidatively damaged guanine base, 8-oxo-7,8-dihydroguanine (OG), via base excision repair (BER). Unrepaired DNA damage may cause disease-inducing mutations, including changes in transcription regulation or a resultant translated protein. When germ-line mutations occur in the gene of the human homolog, MUTYH, it can lead to a condition called MUTYH associated polyposis (MAP), a colorectal cancer (CRC) subtype. Some variants associated with MAP exist in the interdomain connector (IDC) that joins the N-terminal domain responsible for catalysis to the C-terminal domain necessary for OG recognition. The Q324H variant is located in the IDC in a critical binding domain of several downstream repair proteins. Initial studies with this variant demonstrated reduced OG:A repair in vivo yet maintains wild type (WT) activity in vitro. In this study, we employ the mouse Mutyh homolog of Q324H, Q306H, to better characterize Q324H activity due to difficulties in assessing human MUTYH activity in vitro. Although these studies with the mouse homolog variant may also show WT OG:A repair in vitro, due to the location of this variant in the IDC, it’s CRC association may be caused by perturbations in downstream repair.

Investigating the spatial and temporal response of tomato fruit to oxidative damage during postharvest chilling injury

Student Name: Gurpaul Basra
UCD Department: Department of Plant Sciences
UCD Mentor: Dr. Diane Beckles

One of the most frequently used and effective methods to preserve postharvest produce is low temperature management. Refrigeration of fruits and vegetables is beneficial as it can prolong shelf life as well as maintain the overall quality of the product; however, tropical and subtropical produce often do not reap the same benefits as they are not suitable for lower temperatures. These fruits, vegetables and herbs are negatively impacted by low temperature storage, a disorder called postharvest chilling injury (PCI). Tomato fruit represents an important part of human diet, and is also susceptible to PCI, which manifests as a series of physiological, biochemical and molecular changes, such as alterations in the  redox state of the cell (reactions involving the transfer of electrons) and the production of reactive oxygen species (ROS). The accumulation of ROS to toxic levels is harmful to the cell and can accelerate its overall rate of deterioration. Although PCI has been studied for more than 200 years, the development of oxidative damage caused by ROS has not been extensively characterized in the different tissues that compose a tomato fruit. This article will investigate  two different parameters associated to oxidative stress during postharvest cold storage in the columella and pericarp tissues of cherry tomatoes.

Localization of Plant Pathogen Effectors and Plant Innate Immunity

Student Name: Hilal Morrar
UCD Department: Department of Plant Biology
UCD Mentor: Savithramma Dinesh-Kumar

The world loses some food crops to pathogenic infections, and investigating how pathogens infect plants would help reduce how much crop is lost. In turn, this leads to the development of new pesticides and pathogen resistant plants. Plants have two main defense systems, which are Pattern Triggered Immunity and Effector Triggered Immunity. The first system includes the plant’s receptors on the outside that trigger the immune response after detecting a pathogen effector. These extracellular receptors act like plants first layer of defense against invading pathogen. Pathogens have evolved sophisticated means to compromise this primary layer of defense by delivering effector protein inside the plant cells. Plant employ a second layer of defense through intracellular Nuclear Binding – Leucine Rich Repeat (NLR) receptors that direct the Hypersensitive Response after detecting an effector. In nature, these effectors are injected into a plant host cell via the Type Three Secretion System. The secretion system complex protrudes out of the bacterial cells and is comprised of a transmembrane base and a long, narrow needle to directly release effectors into a host. Here we explore the localization of these effectors using confocal microscopy. We engineered effector proteins fused to a fluorescent protein by molecular cloning, resulting in a chimeric protein that was delivered into tobacco leaves through Agrobacterium mediated transformation.  Our data reveals that the pathogen effectors localize to the plasma membrane, chloroplast, and mesophyll cells of the tobacco plant leaves.

Biocatalyst Discovery for the Chemoenzymatic Synthesis of Lacto-N-Tetraose

Student Name: Ivy Tang
UCD Department: Department of Chemistry
UCD Mentor: Xi Chen

Researchers desire an affordable synthetic route for forming human milk oligosaccharides absent in cow milk to study their functions and incorporate them into infant formula. These compounds are believed to provide the protective properties of human milk for preterm infants from necrotizing enterocolitis and other diseases. Many of these compounds can be synthesized with purified enzymes and substrates or microbial cells engineered for their production. However, those containing Galβ1-3GlcNAc linkages, such as lacto-N-tetraose, are generated in low yield and are expensive due to the lack of a encoding β1-3-galactosyltransferase known to express well in suitable hosts. Eight genes encoding β1-3-galactosyltransferases were unsuccessfully cloned into pET-22b(+), an E. coli expression vector. If successful, the vector would be transformed into E. coli BL21(DE3) cells. Enzyme expression titers, optimal reaction conditions, and catalytic properties of the β1-3-galactosyltransferase will be determined. The most suitable enzyme will then used in the total enzymatic synthesis of lacto-N-tetraose from lactose with excellent yield. Affordable synthetic procedures for the preparation of human milk oligosaccharides such as lacto-N-tetraose may lead to improved baby formulas that reduce infant mortality rates.

The Significance and Misforecast of the 15 May 2018 New England Severe Thunderstorm Outbreak

Student Name: Jacob Feuerstein
UCD Department: Land, Air, and Water Resources
UCD Mentor: Dr. Kyaw Tha Paw U

The severe thunderstorm outbreak of 15 May 2018 devastated large portions of the Northeast with huge swaths of damaging wind up to 110 mph and a dozen tornadoes up to EF2 in strength. Five people were killed and 1.4 billion dollars in damage were caused by the event, making it among the costliest and deadliest severe thunderstorm outbreaks in the history of the Northeast. However, the storm prediction center (SPC) failed to forecast the severity of the event in the days before it occurred, potentially leaving many residents of impacted areas uninformed of the dangerous conditions soon to occur. This case study attempts to answer both why this event was relatively severe and why the SPC failed to forecast the event’s severity. This was accomplished by analyzing synoptic conditions, significant mesoscale events, and model data associated with SPC forecasts. The outbreak was also compared to a previous significant Northeastern outbreak that occurred on 31 May 1998. The current analysis concludes that the severe weather event was an unusual combination of an elevated mixed layer and very high wind shear, due to a split stream pattern (in which the jet stream branches into two different components) and a 700mb trough over the West Coast. This is a synoptic setup that matches the one in 1998 very closely. Model and NWS text data suggests that ample evidence of incipient severe weather existed preceding the event, implying that the SPC could have forecast its severity. It was also determined that the event should be classified as a derecho using procedures outlined in Johns and Hirts, 1987. The split stream setup, when taken with its similarity to past significant severe outbreaks over the Northeast, introduces a better understanding of Northeast severe weather that can aid in forecasting of severe thunderstorm potential.

GESI Constructs in PDGFR-N-ARCLIGHT Scaffold

Student Name: Janet Han
UCD Department: Department of Biochemistry and Molecular Medicine,
UCD Mentor: Lin Tian

The recent increase in the number of behavioral diseases with neurobiological bases has increased the effort to find biosensors to monitor neural connections in the brain. Neural biosensors may be used to monitor brain disease progression by tracking neural activity. Current neural biosensors, such as the genetically encoded calcium sensors (Li, Q. et al.) lack the ability to show how frequently the neuron is firing because calcium levels do not have perfect correlation to neural activity. However, neural activity can be tracked by monitoring the activity of potassium ion channel proteins because action potentials rely on these channels to re-polarize the membrane. By cloning a specified peptide (termed GESI, genetically encoded small illuminant) into the potassium ion channel (Kv2.1) protein that activates the dye bromocresol purple (BCP) upon exposure a biosensor that can track neural activity can be created. The peptide will be cloned in an area that will only be exposed to the BCP in solution when the potassium ion channel is active. As a pilot experiment, the Tian Lab at UC Davis has identified several GESIs that can be cloned into PDGFR-N-Arclight, a smaller and easier to manipulate voltage sensitive protein. While this experiment does not directly clone GESIs into the Kv2.1 voltage-gated potassium channel, the PDGFR-N-Arclight scaffold does provide the possibility of developing a voltage-sensitive biosensor used for tracking neural activity due to its similarities to Kv2.1. Furthermore, the PDGFR-N-Arclight scaffold provides insight into how to achieve a Kv2.1 based biosensor.

Metabolites Produced from Bifidobacterium Consumption of Breast Milk Oligosaccharides Suppress the LPS Induced Inflammatory Response in Macrophage Cells

Student Name: Jason Own
UCD Department: Anatomy, Physiology & Cell Biology
UCD Mentor: Dr. Helen Raybould

Macrophages play a key role in maintaining adipose tissue homeostasis, yet recently these cells have become extensively associated with adipose tissue inflammation upon the development of obesity, a disease that affects 2.1 billion people in the world today. In response to the chronic lipid overloading of adipose tissue during the development of obesity, adipose tissue macrophages increase in number and change their localization. This newly attained knowledge shifts medical approaches away from targeting the inflammatory traits of adipose tissue macrophages, towards targeting their metabolic programming. Mediations directed at increasing metabolic capacity might be used to reprogram macrophage metabolism, allowing macrophages to better deal with metabolic challenges during obesity, in order to maintain adipose tissue homeostasis. Taking this into account, the aim of this study was to determine which metabolites produced by Bifidobacterium grown on specific milk oligosaccharides will suppress inflammation induced by the inflammatory stimuli, LPS, in macrophage cells. RAW-Blue cells, a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice, were cultured and exposed to various Bifidobacterium supernatants produced from Bifidobacterium consumption of lactose, bovine milk oligosaccharides, 2′-Fucosyllactose, and 3′-Sialyllactose. These cells were then induced with LPS, activating the transcription factor, NF-kB. Upon activation of NF-kB in these RAW-blue cells, a secreted embryonic alkaline phosphatase was secreted into the cell supernatant. After overnight incubation, a QUANTI-blue macrophage inflammatory assay was executed in order to detect the levels of alkaline phosphatase. This study provides the first evidence that high concentrations of lactose and 2-FL metabolites were found to significantly decrease NF-kB activation. Therefore, it was concluded that metabolites produced from Bifidobacterium consumption of lactose and 2-FL suppress the LPS induced inflammatory response in adipose tissue macrophages.

Volutionary history and genetic basis for elongated posterior spiracles in cactophilic Drosophila species

Student Name: Jerry Liu
UCD Department: Population Biology, Evolution and Ecology
UCD Mentor: Dr. Susan Lott

Developmentally constrained traits are robust to perturbations, including genetic change, environmental change, and internal noise, likely because variation may be detrimental to the organism. However, the interactions between genetic and environmental perturbations and developmental variation are poorly understood. Can traits that are essential for the proper development of the organism change over evolutionary time? If so, do they change in small frequent steps or rare big jumps? In this paper, the evolutionary history of the posterior spiracles in twelve cactophilic Drosophila species spanning about 20 million years of evolution is examined. Position of each abdominal segment was determined at the 1st instar larval stage from each of the species using denticle belts as a proxy. The positions of the denticle belts were measured in proportion to the entire larval body length and analyzed using linear models. The results show that robust traits do evolve, as there are small but significant changes in body plan within the group of cactophilic species, and larger differences between cactophilic and non-cactophilic species. All cactophilic species examined in this paper were shown to have significantly longer posterior spiracles than that of the outgroup species, suggesting that this trait may have arisen as an adaptation to the desert climate. Finally, when comparing the segment position of each species to the mean segment position, it was found that closer segments tend to shift together, which suggests that the genetic changes underlying the observed segment position changes may lie in the late embryonic stages.

Effect of Differences of Shell Thickness and Pressures on the Cracking of Grapes

Student Name: Justin Semelhago
UCD Department: Department of Plant Sciences
UCD Mentor: Kenneth Shackel

Grape splitting has affected the grape and wine industry greatly. Thousands of dollars are spent yearly to cover the cost of grapes that split because they cannot be sold to the consumers or wine companies. Past research only indicates that cracking could be a cause of unevenness of skin surface or the irregularities of pressures within the grape. To inhibit cracking, grapes were spray-painted with white dots randomly on the surface and were then placed in water with randomly spray-painted white dots and analyzed over time. The analysis process consisted of running the images taken of the grapes through MATLAB to produce eigenvalues. The eigenvalues track the percent movement of a centroid of three white dots over time. The eigenvalues were then plotted using SAS and patterns were drawn from the graphs. It was found that eigenvectors in the cracking area were growing in a nonuniform way the moment before the crack. The eigenvectors would become larger in both directions indicating an increasing strain adjacent to where the crack would eventually form. Eigenvectors that were not directly beside the crack but in the cracking area showed a decreasing strain perpendicular to the crack. However, within the same eigenvector there would be an increasing strain perpendicular to the crack. This is caused by the crack pushing the centroids of the dots closer to other dots on the same side of the crack which could be a result of either differences of pressure in that area or an unevenness of skin thickness. 

The Location of Mesoporous Silica Nanoparticles Over Time in Mouse Lungs Following Acute Inhalation

Student Name: Krysta Zmich
UCD Department: Center for Health and the Environment
UCD Mentor: Kent E. Pinkerton

Mesoporous silica nanoparticles (MSNs) are a new and effective method of drug delivery in the body, with the ability to be tailored for continuous or triggered drug release. Past research has shown MSNs to be highly efficient at targeting specific cells in the body, with greater specificity  than chemotherapy treatments for cancer. Previous research has also shown that once MSNs are taken up by the body through inhalation, the particles do not leave the lung one (1), seven (7), and twenty one (21) days after exposure. To identify the location of MSNs in the lung once inhaled, and the potential consequences of these MSNs on the body, immunofluorescent staining of the lung epithelium coupled with confocal scanning fluorescence microscopy was done to identify the precise nature of MSN uptake and retention in the lungs over a period of 21 days post-exposure. Researchers at the Center for Health and the Environment at UC Davis had previously exposed mice (n = 53) to MSNs and removed the lungs for histological sampling. These lung samples were embedded and sectioned for staining and subsequent confocal microscopy imaging. To determine the cell type associated with MSNs, staining with antibodies for CDH1 (epithelial cell surface protein), CD11c (macrophage and dendritic cell surface protein) and Siglec F (macrophage specific surface protein) was performed. No MSNs were found to be present in the epithelial cells of the lung, but instead were found in cells outside of the epithelium and in the airspaces of the lungs. This suggests MSNs are most likely uptaken by cells of the immune system, macrophages or dendritic cells. Future investigation is needed to confirm this theory

Characterization of Breast Cancer Cells Growing on Mitochondria-Driven Metabolism

Student Name: Lindsay Tao
UCD Department: Department of Molecular Biosciences
UCD Mentor: Cecilia Giulivi

Intra-cell heterogeneity in breast tumors may confer resistance to conventional cancer treatments. Certain types of cells may be resistant to a certain treatment, thus allowing cancer to persist. We hypothesized that the heterogeneity of  cancer cells can be evidenced in multiple ways, including the identification of discrepancies in their metabolic processes. To this end, we sought to select and characterize a subpopulation of cells that only utilize mitochondrial metabolism to obtain energy from fuels. Adenocarcinoma breast cancer cells (MDA-MB-231) were grown in a galactose or glucose media. The cells in galactose were selected for the ability to use mitochondrial metabolism, while the cells in glucose served as the control. Cell proliferation, phosphorylating capacity, and markers for differentiation, pluripotency, and metastasis were evaluated in cells grown in galactose versus those in glucose. Cells grown in galactose (cells that rely on mitochondrial metabolism including fatty acid oxidation) were found to proliferate slower, have higher ATP production, and were less pluripotent and less differentiated. Thus, breast cancer cells which were forced to generate ATP from mitochondria displayed unique morphological and metabolic characteristics that seem to include a transition towards stemness.

Studies Towards a Synthesis of 4,10,15-(1-4)tristetrafluorobenzena- 1,7-diazabicyclo[5.5.5]heptadecaphane, a Potential Macrobicyclic Fluoride Receptor

Student Name: Matthew Nemeth
UCD Department: Department of Chemistry
UCD Mentor: Mark Mascal

The detection and monitoring of chemical levels in bodily and environmental systems is a growing objective in the field of synthetic chemistry, following the increasing ability of scientists to synthesize specific chemical receptors. Most artificial anion receptors target phosphate ions or a range of the larger halides: chlorine, bromine, and iodine. This report describes progress in the synthesis of a macrobicylic molecule as well as a macrocyclic intermediate that could act instead as a fluoride receptor. The feasibility of synthesis of 1,4-Bis(carboxymethyl)-2,3,5,6-tetrafluorobenzene 1,4-Bis(cyanomethyl)-2,3,5,6-tetrafluorobenzene were verified with moderate yields, but more time is needed to synthesize 1,4-Bis(ethylamine)-tetrafluorobenzene and carry out the full macrocyclization.

Comparing and Contrasting the Aspects of PCR and Taqman Assays

Student Name: Mehr Sahota
UCD Department: Mouse Biology Program Center for Comparative Medicine
UCD Mentor: Dr. Joshua Wood

A human and a mouse both have 3.1 billion base pairs which makes a mouse’s genome ideal for genetic research (Importance of Mouse Genome 2017). This project will include the comparison of two important pieces of biotechnology: Taqman assays and traditional PCR, in order to discover which is more cost efficient and accuracy of the screening methods when looking DNA insertions. Analyzing these screening accurate will help provide MBP with the most effective way to produce transgenic and Knockout mice thus furthering scientific research. The Taqman assay and traditional PCR will screen a large Knock in of GFP, a Knock in SNP (single nucleotide polymorphism), and a knock in of loxP/CRE which is a site specific recombinase consisting of around 30 base pairs (Valenzuela et. al., 2003). A Taqman assay is initially more expensive, but it is hypothesized to perform better than traditional PCR when working with small changes in the number of base pairs (Redig 2014). On the other hand, traditional PCR is better suited to find a large insertion than a Taqman assay (Valenzuela et. al., 2003). To investigate whether a Taqman assay or PCR is more effective in certain screening methods, time measurements and cost calculations must be taken into consideration for each of the methods. The quality of the screening must also be taken into consideration to ensure that quality is not being sacrificed for cost or time efficiency.

Thermal and kinetic characterization of β-glucosidase B mutants reveal structural and functional relationships

Student Name: Michelle Tong
UCD Department: Department of Chemistry
UCD Mentor: Justin B. Siegel

Predictive algorithms for mutant enzyme activity are important for facilitating enzyme selection in industrial projects. But, due to the lack of a large data set collected on mutant enzymes, current Rosetta programs are not accurate in predicting the kinetic activity and thermal stability of mutants. In order to expand the data set documenting mutant activity, the effect of BglB point mutations on its Michaelis-Menten constants and its thermal stability was looked at in this study. The research revealed an increase in the thermal stability of point-mutations when hydrogen bonds were added and a decrease in catalytic efficiency when packing around the side-chain was increased. By characterizing mutants based on their kinetic activity, binding affinity, thermal stability, and catalytic efficiency, this quantitative data will advance understanding of how enzyme structure relates to function.

Effects of Acetaminophen on Proteasome Activity in Cardiac Cells

Student Name: Mitra Pourmehraban
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Aldrin Gomes

The use of nonsteroidal anti-inflammatory drugs (NSAIDs) which are commonly prescribed to patients as pain relievers have recently been associated with cardiovascular disease (CVD). Acetaminophen (Tylenol), which is not an NSAID but has similar pain relieving effects, is not known to induce CVD. The effects of acetaminophen on proteasome activity in H9C2 rat cardiomyocytes was investigated. Proteasomes are enzymes in the body that degrade intracellular protein. These proteasomes are essential for the body to function. The known effects of acetaminophen include liver damage, but the effects of the drug on the heart have not yet been studied. Using rat cardiac cells and proteasome assays acetaminophen was found to have no significant effect on the proteasome proteolytic activity of all three substrates. These results suggest that acetaminophen is not likely to cause as significant an increase in the risk of developing cardiovascular disease as NSAIDs.

RNA Sequencing data analysis between two of Brassica napus F1s

Student Name: Mizuki Kadowaki
UCD Department: UC Davis Genome Center Plant Biology
UCD Mentor: Julin Maloof

Climate change has deemed it necessary for various crops to adapt to the fast changing environment in order to produce yields. To increase profit, Brassica napus plants must be bred to not only produce more oil but to be less susceptible to climate change. In order to study the gene expression of Brassica napus and apply that information to breeding, RNA sequencing data were analyzed for two different F1 progenies, F1 414 and F1 415, which were created by swapping maternal and paternal parent. Four different tissue types at different stages of the plant’s life were studied. Expression analysis was conducted between these two F1s to see whether genes were differentially expressed between F1 414 and F1 415 throughout the plant’s lifetime and what biological functions are differentially expressed. Single Nucleotide Polymorphism (SNP) identification (allele analysis) between the two F1 generations were correlated with the expression analysis to confirm that gene expression is affected by specific alleles inherited from maternal and paternal parent plants.

Tracking the dynamics of mesoporous silica nanoparticles using fluorescent immunohistochemistry and confocal microscopy

Student Name: Raj Ajudia
UCD Department: Center for Health and the Environment
UCD Mentor: Dr. Kent Pinkerton

Mesoporous silica nanoparticles (MSNs) could possibly become one of the best vehicles for drug delivery created to date. The unique characteristics of MSNs allow for the packaging of medicines into miniscule particles that, once inhaled, allow drugs to be accurately and efficiently delivered to deeper-than-ever target sites in the lung. Much is still unknown about the long-term dynamics of deposition of MSNs in the lungs following initial exposure. In efforts to close this knowledge gap, lab mice were exposed to MSNs for 5 hours, andlung tissues were collected at 1, 7, or 21 days post-exposure. These tissue samples were collected andprepared for examination in a process called fluorescent immunohistochemistry. In this study, epithelial cells in the lungs were tagged with primary antibodies binding to CD11C—e-cadherin proteins. Once fluorescent secondary antibodies and DAPI stain were applied to the tissues, the epithelial cells, alveolar macrophages, and auto-fluorescent MSNs were seen using confocal microscopy. The images confirmed that MSNs were located in and surrounding the alveolar macrophages; however, they were not found in the epithelial cells. Future research may utilize these same methods to search for MSNs in other possible cells of interest in the lungs, such as dendritic cells.

Kinetic and Thermal Characterization of β-glucosidase (BglB) Mutants to Evaluate Effects on Michaelis-Menten Constants and Thermal Stability

Student Name: Raushun Kirtikar
UCD Department: Department of Chemistry
UCD Mentor: Dr. Justin Siegel

Advents in technology are leading to a revolution in the biological research industry, particularly the field of molecular medicine and genomics. A major goal of these advancements is the creation of a predictive software that will determine what effect an enzymatic mutation will have on the protein and the potential applications of such a mutation. The manipulation of proteins has applications ranging from pharmaceuticals to biofuels to wine-making. Using a design-build-test-learn setup, point mutations were first chosen using the computational software, FoldIt. Then, the mutant DNA was inserted into E. coli cells. The protein produced by the cells was then kinetically characterized to determine Michaelis-Menten constants and analyzed for thermal stability by observing the melting temperature. The results of the kinetic and thermal assays did not match the predictions, likely due to the inaccuracy of the current predictive software. This supports the fact that there is a need for more data collection in order to develop a better predictive software.

Predicting Tyrosine Sulfation Sites in Pattern Recognition Receptors

Student Name: Ria Arora
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Grace L. Rosenquist

Tyrosine O-sulfation, a post-translational modification, plays a crucial role in cell signaling and protein-protein interactions. This phenomenon was demonstrated to be essential in the binding of some pattern-associated molecular patterns (PAMPs) to pattern recognition receptors (PRRs). PRRs activate downstream signal cascades, which generate the innate immune response. However, it is not known whether PRRs are tyrosine O-sulfated. In this study, computational methods were used to predict tyrosine O-sulfation sites in PRRs by using a position-specific scoring matrix to examine the similarity of tyrosine sites in PRRs to that of known sulfated sites. Tyrosine O-sulfation was predicted to exist in 57 PRRs observed, with 130 positive sites found. Furthermore, five predicted tyrosine sites were conserved between Toll-like receptors, an important family of PRRs. Predicting tyrosine O-sulfation sites in PRRs could elucidate the prevalence of tyrosine O-sulfation in the innate immune response and assist with developing drugs that target innate immunity.

Sensitivity and Specificity of the PSSM Tyrosine O-Sulfation Calculator

Student Name: Richard Cheng
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Grace L. Rosenquist

One significant but still not completely understood modification of proteins is tyrosineO-sulfation. Because it is expensive and time-consuming to detect O-sulfated tyrosines, computational tools like the position-specific scoring matrix (PSSM) and the Sulfinator have been developed to predict tyrosine O-sulfation. Using a documented set of tyrosines, both sulfated and unsulfated, the sensitivity, specificity, accuracy, and Matthews Coefficient of computational tools for predicting tyrosine O-sulfation were evaluated at various cutoffs. Although the PSSM was found to have relatively high sensitivity and specificity, Jackknifed resampling demonstrated the need for additional O-sulfated tyrosines to be identified to improve the PSSM further. The Sulfinator was found to have lower sensitivity, specificity, accuracy, and Matthews coefficient than the PSSM, and thus is not recommended to be the primary tool for computational studies of tyrosine O-sulfation.

Effects of Motor Oil Extracts on Medaka Fish

Student Name: Shelley Jersey
UCD Department: Department of Anatomy, Physiology, and Cell Biology
UCD Mentor: Dr. Swee Teh

Oil from motor vehicles can leak onto roads. This oil is flushed into nearby water bodies when it rains, potentially harming aquatic life. The purpose of this study is to determine the effect of various dilutions of motor oil extracts on the embryonic development of Japanese Qurt Medaka fish (Oryzias latipes). The first batch of embryos was exposed to four different dilutions of motor oil extract and a field sample obtained from the UC Davis VP56 parking lot. A second batch was treated with 0.01% dimethyl sulfoxide (DMSO) to increase the permeability of the embryo’s chorion and subsequently exposed to an undiluted extract and four different dilutions of motor oil extract. All embryos were placed in a 96-well plate with 200 uL of their respective dilutions. Both batches were monitored for changes in morphology, delayed growth, and mortality. Some deformities were observed in the embryos but did not seem to be consistent with any particular group. However, embryos exposed to the field sample and the undiluted extract showed delayed growth and high mortality rates. The groups exposed to higher concentrations of motor oil extract seemed to have higher mortality rates. The data suggests that motor oil extracts may be harmful to aquatic life, but further studies and analysis would need to be conducted to determine if an imminent threat exists.

X-ray Induced Förster Resonance Energy Transfer between Quantum Dot-Nanoporphyrin Conjugates to Generate Singlet Oxygen

Student Name: Susan Garcia
UCD Department: Department of Chemistry
UCD Mentor: Dr. Ting Guo

X-rays combined with Förster Resonance Energy Transfer, can produce singlet oxygen, a reactive oxygen species (ROS), through the conjugation of Quantum Dots encapsulated in Nanoporphyrins. This type of ROS can be used in health applications such Photodynamic therapy and Photothermal therapy. Enhanced singlet oxygen through this process can progress towards a new type of treatment for tumors that will only eradicate malignant cells rather than healthy cells. In this research, a previous experiment was conducted and was found unrepeatable,  presumably, due to material changing over time. The goal of this paper was to attempt to replicate this experiment to present replication and consistency. However, results indicated that the production of enhanced 1O2 is higher when Quantum Dots are encapsulated in Nanopohyrins than not being encapsulated. The replicating experiment presented the enhanced production of 1O2 lower than the first experiment. It is hypothesized SOSG was quenched and that is why the fluorescence signal decreased, however, this is up for further study.  This is significant because the replication of the experiment has added new data that demonstrates X-ray combined with FRET and the conjugation of Quantum Dots produces more enhanced  1O2  than Nanoporphyrin itself. Concluding, that 1O2  is enhanced through combination of X-rays and FRET conjugating Nanoporphyrin and Quantum Dots, leading to alternative treatments for tumor reductions.

Finding the Genetic Diversity of Juglans microcarpa

Student Name: Taoyi Li
UCD Department: Plant Pathology Department
UCD Mentor: Dr. Daniel Kluepfel

Rootstocks are genetically distinct from the scion, the fruit producing portion of the commercial walnut. Rootstocks are generally better suited to the environmental conditions present in the rhizosphere, often providing resistance to soil borne pests that may otherwise affect the scion and decrease walnut production. Regarding the rootstocks of commercial walnut plants, a better rootstock needs to be developed because a greater resistance to soil borne microbes and pests is desired by the walnut industry. The Texas walnut tree Juglans microcarpa has demonstrated resistance to the crown gall disease causing microbe Agrobacterium tumefaciens. To facilitate rootstock development, we evaluated recent USDA acquisitions of J. microcarpa germplasm for genetic diversity. A sample of 272 individual genotypes were analyzed, originating from Texas, Oklahoma, Kansas, and New Mexico. Population structure and genetic diversity analyses were carried out with adegenet and Poppr packages in R using multilocus microsatellite genotypes. Within the sample groups, little population structure was found with principal component analysis. Analysis of molecular variance showed little differentiation in population. Hardy-Weinberg and F-statistics test showed the groups to be in Hardy-Weinberg disequilibrium for most loci.

Studying the Relationship Between the Stretching of a Grape’s Skin and Cracking

Student Name: Trevor Zinky
UCD Department: Department of Plant Sciences
UCD Mentor: Kenneth Shackel

Berry cracking in a grape is a common problem that is unable to be predicted. Grape cracking causes an exposure of the inside of the grape, leading to an increased susceptibility to disease. In this project, existing data consisting of timelapse pictures was studied to determine whether a cause of grape cracking can be predicted based on the stretching of the grape’s skin. Random patterns of spray paint dots were applied to each grape, which were then submerged in water and photographed periodically to capture water-induced absorption and expansion. Each photo was analyzed and positional data was created based on the tracking of each dot, which was then converted into graphical representations. Initial observations revealed that fractures in the dots of paint were observed before cracks in the skin of the grape. Examination of the area change of the dots showed that dots of paint can stretch and increase their area over time without fracturing. Visual observations of dot fracturing also indicated that a crack may form in the skin of the grape before it is visible on the surface in areas without paint dots. These results show that an irreversible state of cracking could occur in the grape far before the point where the crack is visible on the grape’s surface. However, more research is needed to determine if fracturing in the dot occurs at the same time of cracking in the skin of the grape.

Information theory suggests that long introns splice via exon definition in C. elegans

Student Name: Veronica Lee
UCD Department: Molecular & Cellular Biology
UCD Mentor: Dr. Ian Korf

Pre-mRNA splicing, a process in which introns are removed and exons are ligated, is an essential part of gene expression. Revealing more about the process of splicing in eukaryotic DNA can lead to a greater understanding of and control over gene expression, which yields advancements in medicine, health, and food/agriculture. This research project hopes to answer the question: are long and short introns in eukaryotic DNA spliced in fundamentally different ways? In this project, a bioinformatics approach (Unix operating system, text editor Notepad++, and programming language Perl) was used to analyze k-mer distributions of long and short introns and their flanking exons. Results showed that introns similar in length have similar k-mer distributions, while long and short introns have significantly different k-mer distributions. It was also shown that short-flanked exons have k-mer distributions that are more similar to other short-flanked exons than to long-flanked exons. This may indicate different protein binding sites for spliceosomal snRNPs during the splicing of long and short introns, providing further evidence for intron and exon definition.

Organic soils influence tomato resistance to insect attack

Student Name: Victoria Yang
UCD Department: Department of Plant Pathology
UCD Mentor: Clare Casteel

Studies have shown that organic farming techniques applied on soil leads to increased health and microbiota diversity. The optimal management of multitrophic interactions between soil microbial communities, crops, insect pests, and pathogens has the potential to increase plant health and agroecosystem productivity, resilience, and sustainability. However, the underlying mechanisms are unclear, and the impact of soil health-building management practices on pest and virus resistance remains to be quantified in economically relevant crops. Experiments using soil from three field locations in Northern California and from the Russell Ranch Century Experiment investigated the effect of soil health and microbial community composition on processing tomato (Solanum lycopersicum) plant growth and nutrition, induction of defense compounds, and attractiveness to insect vectors. These results support the hypothesis that organic practices decrease pest populations. The findings of this experiment pave the way for the next generation of insect control, leading to a cleaner future in which the natural defense systems of plants are utilized, instead of an over-reliance on chemical insecticides.

Using Algae to Investigate Cytokinesis Conservation

Student Name: Yolanda Shen
UCD Department: Department of Plant Sciences
UCD Mentor: Siobhan M. Brady

Little is known of the details of plant cytokinesis, despite it being one of the fundamental processes in plant biology. Endosidin 7 (ES7), a highly specific probe, has been shown to prevent cell cytokinesis by inhibiting the synthesis of callose in Arabidopsis thaliana. This research utilized Penium margaritaceum, an algae with a cell wall that is structurally and developmentally similar to that of land plants. Immunofluorescent tagging of cell plate-related polymers created an understanding of ES7’s effects on unicellular algae in comparison to complex plants. ES7 was found to affect the size of the isthmus zone, while JIM5-labeled pectin particles were also detected in the isthmus. In this experiment, the phenotypic observations of ES7-treated algae serve as preliminary data for future analysis of the target of ES7. If the target can be discovered, it will indicate the presence of a conserved component of cytokinesis between algae and plants. Ultimately, this research laid a foundation for a comprehensive understanding of the evolutionarily conserved pathways of cytokinesis.

2016
 

Interactions Between Candidatus liberibacter and Potato Virus Y in Russet Burbank Potatoes

Student Name: Alex Kuang
UCD Department: Plant Pathology
UCD Mentor: Dr. Clare Casteel

Liberibacter candidatus (LSO) is a bacteria causing extensive damage to potatoes. Potato Virus Y (PVY) one of the most prominent viruses affecting all potato crops grown in America. The insect vectors for both pathogens are often found on the same harvest at different times, and this study seeks a relationship between LSO and PVY in which one may be facilitating the other. If such a connection is found, treatment for one pathogen may remedy the other as well. Further research may lead to a lesser need for pesticides and an increased crop yield.

Examining Automated Methods of Boxing in EMAN for 3-D Reconstruction

Student Name: Alexander Xu
UCD Mentor: Dr. R. Holland Cheng

Selecting individual virus particles in micrographs, or boxing, is done frequently by hand, but the automation of this process would allow for greater efficiency during the process of 3-D reconstruction. Two methods of semi-automated boxing, autoboxing and autoboxing from references, were tested, both of which are two automatic boxing settings found as a part of the program Electron Micrograph Analysis (EMAN), version 1.9. Autoboxing from references appears to pick up slightly less undesirable particles, each of which were reviewed manually after automatic selection. After boxing, selected virus particles can then be used to form a 3-D reconstruction of the virus, whose accuracy is dependent on the quality of particles selected, and help with both virus analysis and virus-like particle (VLP) creation. Particles selected from autoboxing from references, however, appear to form less accurate reconstructions of viruses after 3-D reconstruction and 10 cycles of refinements.

The Synthesis of 2,3,4,5,6-pentafluorobenzene Diazonium Tetrafluoroborate and its Possible Function as a P-type Dopant=

Student Name: Alexandra Li
UCD Department: Chemistry
UCD Mentor: Dr. Mark Mascal

Recently, organic semiconductors have gained more attention as an alternative to inorganic semiconductors. Organic electronics are lighter, more flexible, and more low-cost than inorganic ones and have many possible applications, including thinner OLEDs (Organic Light Emitting Diodes) and cheaper solar cells. Organic polymers can become more efficient with the help of a more efficient dopant, an agent that allows a semiconductor to conduct current by either adding an isolated electron (n-type) or creating a hole in a sea of electrons (p-type). The goal of the project was to create an electrophilic diazonium salt in order to test whether it will be a more efficient p-type dopant than standard ones. A new procedure to make the diazonium salt was created, which led to successful synthesis of the compound, PFBDT. Critical data of the general chemical properties of PFBDT was provided to chemical engineers, who will perform further analysis of the compound’s role as a dopant.

Measuring and Predicting Syringe Hub Loss

Student Name: Allison Mayes
UCD Department: Mouse Biology Project
UCD Mentor: Dr. Kristin Grimsrud

The design of a conventional syringe includes a dead space between the needle and barrel of the syringe called the hub. This space collects liquid that once drawn up, cannot be plunged out. As a result, the substance remaining in the hub is inevitably wasted and difficult to account for. Little information is known about the true amount of hub loss that is expected from a given vial of controlled substances. We calculated and measured the hub volume using a variety of different syringes and vial volumes to determine the anticipated substance loss. Additionally, we tested the effects of interpersonal variability, number of syringes used per vial, volume drawn up in each syringe, and other real world conditions. Prior to finding the hub loss per vial, we calculated the hub volume of three different syringe and needle types. The two conventional 1ml syringe types used had means of 0.068ml and 0.066ml of hub loss. The hubless tuberculin syringe had a mean of 0ml of hub loss. After calculating the hub volume of each syringe, we tested expected hub loss from 10ml and 20ml vials. The trend of these tests showed an increase in hub loss as the number of syringes used increased. When 50 syringes were used to draw out half of a 10ml vial, there was a mean of 3.3ml of hub loss. When 10 syringes were used to draw out half of a 10ml vial, there was a mean of 0.5ml of hub loss. Of the recorded hub loss, the range of all datasets ranged from 0.1 to 2.9. These wide ranges display inconsistent amounts of hub loss per vial. The inconsistency may have been due to real world variables such as imprecise starting vial volumes, interpersonal variability, air bubbles, and unstable pressure within vials. The aim of this study was to establish a reasonable range of hub loss to monitor the usage of controlled substances. Based on our data, a reasonable range of hub loss depends on the number of syringes used to draw from a single vial, with the volume per draw not significantly affecting hub loss. As an example, a reasonable amount of hub loss from a vial when 10 syringes are used is 0.1ml-1.07ml. When 50 syringes are used, 1.99ml-4.53ml. The study may also be used to improve laboratory protocols so hub loss can be minimized.

A Study on the Effectiveness of Different Carbon Sources on Anaerobic Soil Disinfestation (ASD) and Characterization of Soil Fungi Associated with ASD

Student Name: Anne Chamberlain
UCD Department: Plant Pathology
UCD Mentor: Dr. Daniel Kluepful

Anaerobic soil disinfestation (ASD) is an environmentally sustainable alternative to soil fumigation that has been shown to be effective for controlling a broad range of soil phytopathogens, including the crown gall causing bacterial walnut phytopathogen, Agrobacterium tumefaciens. This research focuses on specific aspects of the ASD process, particularly on evaluating a variety of different agricultural waste products as carbon sources (C-sources), and using culture and molecular-based methods to identify microbes present in ASD treatments. 

Developing CAPS Markers for Genotyping Sunflower Mutant Populations

Student Name: Benjamin Caswell
UCD Department: Plant Biology
UCD Mentor: Dr. Stacey Harmer

While a great deal is known about the effects of plant circadian clocks on general biological functions, the molecular mechanisms behind this regulation are somewhat uncertain. Sunflowers with known point mutations were obtained, primers were designed, and restriction enzymes selected for each mutant. The primers were tested on wild-type DNA to determine whether they effectively amplified DNA fragments around the desired point mutations. Since the mutations are in genes associated with circadian clocks, this work will help facilitate experiments in this field in the future.

Exploration of a New Barium Antimony Selenide Compound as a Thermoelectric material

Student Name: Chang Hwan Kwak
UCD Department: Chemistry
UCD Mentor: Dr. Kirill Kovnir

Study in the field of thermoelectricity, the conversion of heat energy into electrical energy and vice versa, He explored a new barium antimony selenide compound, Ba6Sb7Se16.11, that has been previously determined to have a high potential as a thermoelectric material. Bryan became the pioneer to study the thermal conductivity, resistivity, and Seebeck coefficient of the barium antimony selenide compound. These components are required to calculate the figure of merit, which determines how good a thermoelectric material is. He synthesized the compound via high-temperature solid-state reactions, using a furnace and a Spark Plasma Sintering (SPS) device. With a Rigaku-600 Miniflex powder X-ray diffraction machine, the phase of the compound was determined. Bryan hopes to continue his research by keeping in contact with his mentor for further results. 

Understanding Crop Immunity for Increasing Crop Yields

Student Name: Christian Mojica
UCD Department: Plant Biology
UCD Mentor: Dr. Dinesh-Kumar

In order to defend itself from various pathogens, plants use various methods including ROS secretion by RbohD in order to eliminate pathogens. Various signaling cascades and protein interactions can lead to ROS secretion, yet only few have been studied. Some of the unstudied protein interactions are the ones between RLCKs and RbohD, yet here we provide this information through a method known as yeast two hybrid. These findings will highlight the direct roles that RLCKs play in ROS secretion.

Determining fertility in Mus musculus eggs for pronuclear injection should be optimized to prevent wastage

Student Name: Darren Tong
UCD Department: Mouse Biology Project
UCD Mentor: Josh Wood

The new CRISPR-Cas9 system, when combined with pronuclear injection, is capable of inducing specific mutations in mice. However, this process has a low mutation rate, around 15% [1]. The aim of this study is to determine if alterations to the timing of the pronuclear injections will improve the aforementioned mutation rate. Based on the results, it seems that around 32.8% of eggs thought to be unfertile are actually fertile. Because they are not used, the embryos are disposed of. This leads to a large amount of waste, and more importantly, a large amount of mice needed. These mice must be fed, and taken care of. Using embryos efficiently means that fewer mice will be needed as a lower total of eggs will be necessary. 

Distinguishing C. elegans chromosomes through amplicon size polymorphisms

Student Name: David Wang
UCD Department: Molecular and Cellular Biology
UCD Mentor: Dr. Ian Korf

The worm C. elegans is a transparent, free-living nematode that has shown to be an efficient and effective model organism. Since 65% of human disease genes have homologues on the C. elegans genome, it has great potential to be an invaluable research organism in the health and biomedical sciences. Researchers studying the C. elegans worm, however, have no cost-effective method of distinguishing homologous chromosomes. This becomes a problem when investigating its reproductive processes such as chromosomal loss in aneuploid individuals. In our project, we propose a cost-effective method of distinguishing homologous chromosomes through size differences on specific regions on the genome which are the result of indel polymorphisms. These size differences can be amplified through PCR and distinguished through gel electrophoresis. We analyzed a dataset of all indel polymorphisms referenced to the N2 strain on significant number of C. elegans strains. We were able to find six strain pairs which had size polymorphisms on all six chromosomes. Laboratory work confirmed our method as a viable, cost-effective solution to distinguishing homologous chromosomes. Our research aids in better understanding the reproductive processes of C. elegans worms which may increase our capacity to utilize it as an effective research organism.

The Influence of Surface Charge on Engineered Nanoparticle Delivery and Retention in the Respiratory Tract

Student Name: Delaney Buskard
UCD Department: Vet Med
UCD Mentor: Dr. Kent Pinkerton

Mesoporous silica nanoparticles (MSNs) are inorganic-based nanocarriers that have porous channels. Their channels can be filled with medicinal compounds and can be utilized to target various disease of the respiratory system. The purpose of the study is to determine what the MSNs do after being deposited in the lungs following a single, acute period of inhalation. Mice were exposed to aerosolized MSNs or filtered air through a nose-only exposure system for 5.5 hours. They were examined 0, 1, 7, and 21 days post exposure and bronchoalveolar lavage was performed. The lung tissue was sectioned and the bronchoalveolar lavage fluid was used to prepare cytospin slides. DAPI or Hematoxylin and Eosin (H&E) were stains that were used to detect the presence of MSNs or inflammatory or structural changes in the respiratory system, respectively. It was discovered that neither positively or negatively charged MSNs do not cause an inflammatory response in the respiratory systems of mice. It was also discovered that the positive MSNs have a better retention rate than the negative ones do. Over time, the negative ones were being released from the macrophages that engulfed them. Overall, MSNs show potential to be a good drug delivery system. 

Elucidating Maize Diterpene Metabolism: Site-Directed Mutagenesis of Kaurene Synthase 2

Student Name: Eileen Toh
UCD Department: Plant Biology
UCD Mentor: Dr. Philipp Zerbe

As the most harvested crop in the United States, maize (Zea mays) serves as a source of food, livestock feed, and biofuel. Pests, pathogens, and abiotic stresses contribute to significant crop losses, and the stresses, particularly drought, have only been exacerbated by climate change and rising global temperatures. All higher plants including maize produce an array of specialized metabolites that coordinate the plant’s interaction with the environment. Among these metabolites, diterpenes constitute the largest and most diverse group of compounds and have critical functions in the defense of maize against pest and pathogens. Recently, two diterpene synthase (diTPS) enzymes were identified in maize and shown to play a role in these stress responses: kaurene synthase 2 (KS2) and kaurene synthase 4 (KS4). While both proteins utilize the same substrate, ent-copalyl diphosphate (ent-CPP), which is produced by the class II diTPS Anther Ear2 (An2), the two enzymes produce different products. Investigation of the structural-functional differences of these two enzymes through expression in Escherichia coli (E. coli) and activity analysis with gas chromatography/mass spectrometry (GC/MS) enabled the identification of several amino acid residues that are important for the distinct enzyme functions. These findings contribute to our understanding of the chemical diversity of plant diterpene metabolism, and may be an opportunity for improving stress resistance in maize.

Investigating Simulator Sickness during an Immersive Virtual Reality Navigation

Student Name: Elli Stogiannou
UCD Department: Psychology
UCD Mentor: Dr. Arne Ekstorm

Simulator sickness is a condition that has emerged along with the advancements in the field of Virtual Reality (VR). Simulator sickness originates from the incongruity between the motion of the simulation and the motion of the performer in the simulation. Simulator sickness (SS), also known as cyber-sickness, is characterized by multiple symptoms, clustered in three general categories: symptoms related to nausea, oculomotor disturbances , and disorientation (Kennedy, Lane, Berbaum & Lilienthal, 1993). However, there is deficient insight into what causes SS, and how it can be effectively prevented. This research investigates the factors that serve as indicators as to whether someone is susceptible to SS and further explores the possible ways to prevent the appearance of SS related symptoms. Peripheral SS data from a navigation experiment using a head-mounted VR display and an omnidirectional treadmill was coded and analyzed to determine which measures, collected by the experimenters, might be able to predict subsequent attrition in the study. Results obtained show that experience with video games and treadmill proficiency, as rated by the experimenter during a training phase on the VR treadmill, are critical factors that may predict SS. Based on this analysis, if these factors are rated poorly, the probability of SS occurring is high. These findings have major implications for both the private sector, where companies are producing VR for commercial uses, and for the public sector. In the public domain, this research is beneficial for services using VR interfaces for educational purposes, such as Army Institutes, and it contributes to the field of spatial cognition research.

Potential Tyrosine Sulfation Sites in Cadherins

Student Name: Emily Yang
UCD Department: NPB
UCD Mentor: Dr. Grace Rosenquist

Tyrosine sulfation is a permanent posttranslational modification to a protein that regulates protein-protein interactions. This project focuses on predicting potential tyrosine sulfation sites in type II cadherins, transmembrane proteins dependent on calcium-ion binding. Evidence from statistical data, 3-D protein modeling, and conservation data supports that tyrosine sulfation could occur in type II cadherins. Two of the predicted potential tyrosine sulfation sites showed to contain known calcium-binding sites. However, a study has shown tyrosine sulfation to decrease calcium-ion binding in a particular peptide hormone. If this study is true for all proteins, the location of tyrosine sulfation sites near calcium-binding sites would be impractical. Further research would be needed to clarify if tyrosine sulfation actually occurs in cadherins and the effect it might have on cadherin function. 

Determining the role of endogenous auxin in sunflower heliotropism

Student Name: Faith Ajayi
UCD Department: Plant Biology
UCD Mentor: Dr. Stacey Harmer & Hagop Atamian

Sunflowers grow in response to sunlight in a movement called heliotropism, in which they track the sun. Their heliotropism has been proven to be regulated not by direct light stimulus, but by their circadian clock (Atamian et al. 2016). We now show the role of auxin, a naturally occurring plant growth hormone, in the clock’s regulation of sunflower heliotropism. Our experiments suggest that auxin is released at different times during the sunflower’s heliotropic cycle, causing stem elongation, which makes the plant grow in different directions.

Chlorella sorokiniana Growth in Wastewaters

Student Name: Gregory Martin
UCD Department: Chemistry
UCD Mentor: Dr. Annaliese Franz

The algae species Chlorella sorokiniana was grown in dairy waste water (DWW) and in nitrogen reduced anaerobic digester effluent (ADE) at differing concentrations to find the optimal concentrations for growth. The effect the waste water had on the algae was measured by absorbance at 750nm, chlorophyll fluorescence, Nile Red lipid assay, and microscopy. Results indicated that the maximum concentration of DWW Chlorella sorokiniana can grow in while achieving robust growth is 56% and 100% for ADE. The most robust growth and lipid production were seen in the lowest concentrations of DWW and ADE at 10% and 50% respectively. The effect of additional ammonia in ADE on algal growth was also tested and demonstrated that Chlorella sorokiniana will not grow in concentrations over 10mM additional ammonium in 100% ADE.

An Investigation into the Buffering capacity of foodstuffs

Student Name: Henry Harrigan
UCD Department: Biological & Agricultural Engineering
UCD Mentor: Dr. Gail Bornhorst

While many studies have been conducted into the various buffering capacities of feed and cheese, no comprehensive study has ever been conducted into buffering capacity of protein and carbohydrate in the gastric environment. One specific area of interest is into how buffering capacity affects larger scale digestive processes in the stomach, such as gastric emptying and enzymatic secretion. To understand this, we first need a understanding of buffering capacity in the gastric environment. In this study we examined the buffering capacities of a variety of different mixtures of basic carbohydrate and protein. The two constituents were Whey protein isolate, and granulated cane sugar. These were dissolved in water as well as apple juice depending on FDA determined values for what average Americans consume. The difference in the buffering capacity between these repetitions will help us understand how the buffering capacity of foodstuffs changes with secretion of HCL. We found that buffering capacity of whey in water and juice is highly augmented in juice as opposed to water, and is further increased by the addition of whey protein, but little change results from the addition of sugar.

Primary and Tertiary Trends of Tyrosine Sulfation

Student Name: Jack Youstra
UCD Department: Molecular and Cellular Biology
UCD Mentor: Dr. Grace Rosenquist

Tyrosine sulfation is a modification to tyrosine where sulfate ions are attached to the hydroxyl group at the end of the tyrosine. Primary structure trends in sulfated sites were revealed by comparing a sample set of 104 sulfated and 674 unsulfated tyrosine sites while tertiary structure trends were revealed by analyzing x-ray crystallography data from sulfated proteins. Primary structure analysis revealed the linkage between charge, hydrophobic nature, and proximity to a terminus with sulfation as well as the linkage between outliers, such as cysteine, with sulfation. Tertiary structure analysis revealed the linkage between hydrogen bonding and sulfation, especially with positively-charged amino acids. These tertiary characterizations reveal the increased stability brought by the increased hydrogen bonding network from tyrosine sulfation while the primary characterizations help provide refinement to tyrosine sulfation prediction when coupled with the existing PSSM method of prediction.

Comparison of apoE4 and its 61T mutation and lipid-bound stability of apolipoproteins

Student Name: Jacquelin Ho
UCD Department: Biochemistry & Molecular Medicine
UCD Mentor: Dr. John Voss

Amyloid beta (Aβ) is a protein commonly found in the membranes of neurons. Because of its tendency to misfold and oligomerize, the transport of Aβ out of the brain is crucial to cognitive health. The apolipoprotein E (apoE) is responsible for the transport of Aβ; consequently, its gene is the sole genetic cause of Alzheimer’s that has been widely agreed upon. Of the three alleles E2, E3, and E4, the apoE4 protein is the most closely associated with increased Alzheimer’s risk. ApoE4 differs from ApoE3 at the 112th amino acid; in this study, the positively charged 112 arginine of ApoE4 was mutated into a 112 serine to resemble the uncharged 112 cysteine in ApoE3. Six drugs still under development were obtained, and as a result their manufacturers and details must remain confidential; they will be referred to as drugs A through F. To determine whether the effects of drugs A-F on the ApoE4 mutant were significant, the EPR spectra for the E3-like mutant and the E4 clone were scanned and compared. The greatest difference between the E3-like and E4 spectra was found with the drugs D and F, indicating that the drugs may have changed the protein’s structure. To further observe the behavior of apolipoproteins, the stability of lipid-bound nanodiscs of apolipoprotein A with diameters of 7.8 nm, 8.4 nm, and 9.6 nm were observed through comparison of samples in SDS-PAGE gels, EPR, and Coomassie Assays in the second phase of this project. In general, the SDS-PAGE gels showed that the lipid-bound samples retained shape better than their lipid-free counterparts.

Confirming Arabidopsis Mutants Hypersensitive to ES7

Student Name: John Almazan
UCD Department: Plant Science
UCD Mentor: Dr. Georgia Drakakaki

Cytokinesis is an essential developmental process for plants. Currently, scientists have limited knowledge on the processes that occur within cytokinesis. John Almazan is researching the effects of Endosidin 7, a chemical that inhibits the cytokinesis phase of plant cell replication, on Arabidopsis Thaliana mutant seedlings. The primary goal of his research is to confirm whether the mutant seedlings are hypersensitive to Endosidin 7. Studying ES7 and its hypersensitivity effects on the mutant seedlings will confirm that there are additional proteins involved in callose deposition at the plant cell plate. Callose deposition is a significant process for the later stages of cytokinesis when a polysaccharide called callose is deposited at the cell plate, providing mechanical support to the membrane network before the plant cell splits into two daughter cells. Using a software called ImageJ and confocal microscopy, John will monitor the seedlings’ growth and observe ES7’s effects on the plant cells. This research can potentially lead to the identification of the specific proteins that assist the callose deposition process.   

Using a Dairy Relevant Bacterial Mock Community to Compare Quantitative PCR, High-Throughput Sequencing, and Culture-Based Cell Enumeration Methods 

Student Name: Kane Tian

UCD Department: Food Science and Technology

UCD Mentor: Dr. Maria Marco

A study that will contribute toward the improvement of quantification methods for bacterial profiling. This study is comparing high-throughput sequencing, plate counting, and Quantitative PCR using a bacterial community with dairy-associated bacterial species in order to assess whether current methods produce consistent and similar results. Using these three methods, this mock community is also testing the biases introduced by PCR and DNA extraction kits.

Changes in M1 and M2 Macrophage Phenotype Numbers following Long-term Smoking Cessation in Spontaneously Hypertensive Rats

Student Name: Katie Li
UCD Department: Vet Med
UCD Mentor: Dr. Kent Pinkerton

Smoking leads to increased susceptibility to heart attack, stroke, lung cancer, and other respiratory complications, such as chronic obstructive pulmonary disease (COPD), emphysema, bronchitis and persistent cough. COPD is the third leading cause of death in the world today (1). Previous findings on smoker rats show increased epithelial volume, mucin content, and also consistent inflammation in the form of high numbers of macrophages. The purpose of this project was to determine whether macrophage numbers in rat tissues after a period of smoking cessation could be used as a unbiased marker of former smoking. In addition, this study was designed to determine the relative proportion of M1 or M2 phenotype macrophages (markers of active inflammation or lung repair, respectively) in the lungs of old rats as well as former smoker rats as a possible marker of continued lung inflammation or repair. The lungs of 12 male spontaneously hypertensive (SH) rats were examined using histological methods. There were significantly more M1 macrophages found in the tobacco smoke (TS) group compared to the filtered air (FA) group. In the TS group, large numbers of foamy macrophages were observed in the subpleural and terminal bronchial regions. Meanwhile in the FA group, the macrophages found were more dense and scattered throughout the tissue. Findings suggest certain changes in rat lungs due to tobacco smoke are still present after long-term cessation- some changes are irreversible and result in chronic damage. The cellular changes that occur in the cell due to TS exposure are important to consider when designing treatments for those who have stopped smoking. 

Gene Expression Differences Between Fayoumi and Leghorn Chicken and their Impact on NDV Infection Resistance.

Student Name: Kevin Yang
UCD Department: Animal Science
UCD Mentor: Dr. Huaijun Zhou

NDV, Newcastle Disease Virus, is a negative sense single RNA strand virus that infects many wild Avian species. While NDV poses no major threat to humans directly, it causes major losses in the poultry industry and disrupts the poultry food supply. It has been shown that differences in immune response has associated genetic factors that can affect susceptibility or resistance. Determining what those genetic factors are in chicken response to NDV can greatly increase our knowledge of host-pathogen interaction, but also improve genetic selection for resistant birds. And so, the goal of this project was to determine difference in gene expression between two distinct chicken breeds, Fayoumi and Leghorn. Past research has demonstrated that Fayoumi are more resistant to viral infections while Leghorn are more susceptible (Wang, 2014). Chickens were hatched and subjected to heat treatment and viral infection. Samples were collected at 3 different time points: 2 dpi, 6 dpi, and 10 dpi and mRNA was extracted. cDNA was generated and analyzed using qPCR. Analysis of the data demonstrates that Fayoumi gene expression for TLR3, TLR7, IRF7, CD8A in spleen and TLR3, PRKCD, and CD8A in lung is greater when compared to Leghorn. The difference in expression of these genes between these two lines are possibly associated with why Fayoumi has a greater resistance to NDV infection, however further studies must be conducted to show a direct connection between this difference in gene expression and the genetic mechanism used to cause the difference in resistance between Fayoumi and Leghorn. 

Recommended Methods Such As Folding, Recutting, and Bagging Techniques on Leaves to Measure Stem Water Potential in Cherries, Walnuts, Pistachios, and Grapes

Student Name: Lisa Xinying Wu
UCD Mentor: Dr. Ken Shackel

According to the soil-plant-atmosphere-continuum (SPAC) model, water from soil is under tension within a plant, which allows for water to be transported from the root to the leaf of a plant. Water potential is a way of quantifying the tension in the plant, which directly correlates to its water stress. The Scholander pressure chamber was used to test different methods of finding stem water potential in plants and its impact on the measurement taken. This research was done on adjacent leaves, testing for the difference in stem water potential in leaves that were folded and unfolded, and between those cut directly from a branch by its petiole and those with petioles recut after being pulled off a branch. Additionally, research was done on pistachio trees by comparing branches versus individual leaflets, as there is a problem with latex being mistaken for water. Adjacent leaves were placed into light-proof bags and placed into a pressure chamber, where pressure was slowly added until water visibly left the xylem of the leaf. It was found that folding and recutting generally did not have an impact on the measured stem water potential, which will allow farmers to adapt these methods to their convenience instead of worrying about whether or not they need to fold their leaves or cut a leaf only once to fit into the pressure chambers. With pistachio trees, bagging an entire branch produces very close results to bagging just a terminal leaflet, which will allow agriculturalists to pick which method to use that would produce the clearest measure of water potential. In the future, this research can possibly be used to determine a universal and most accurate method of finding stem water potential.

The Effects of Indomethacin on Cardiac Protein Degradation

Student Name: Logan Samuel
UCD Mentor: Dr. Aldrin Gomes

NSAIDs are the most commonly used drugs around the world. They are linked to many side effects such as heart attack and stroke. More specifically, NSAIDs affect the proteasome, a multimeric enzyme that degrades damaged and old protein. Without proteasomes, the body would not be able to function. Indomethacin is a common NSAID used to treat pain, inflammation, arthritis, and patent ductus arteriosis. It has been shown to cause gastrointestinal problems, however, the effects of the drug on proteasomes has not been studied. Due to the high abundance of proteasome activity in the heart, eight mice were injected with indomethacin to investigate the effects of the drug on proteasome activity in the heart. Using western blots, biological assays, and data analysis on homogenized heart samples, it was determined that the control mice and indomethacin injected mice had similar proteasome activity levels. However, trends suggest that proteasomes were being slightly effected by the drug.

Development of UBQ10pro tRFP-WIP1 plasmid for Plant Cell Immunity Studies

Student Name: Maken Horton
UCD Department: Plant Biology
UCD Mentor: Dr. Dinesh Kumar

Plant innate immunology introduces a new plasmid containing a promoter, UBQ10pro, which does not react with 35Spro. Previously, two 35Spro promoters on different plasmids inside the plant arabidopsis co-silenced each other and prevented fluorescent proteins from being produced. This fluorescent nucleic membrane marker protein allows her to see how chloroplasts and stromule extensions interact with the nucleus in the immune responses. Majken transferred the gene tRFP-WIP1 from a vector containing 35Spro to a vector with UBQ10pro. This allows the cell to produce both proteins in large quantities that allow the organelles to be studied. In the future, this research and plasmid will help farmers and agricultural scientist protect and understand plant health leading to increased crop yield and quality.

Analyzing the Role of the Protein Kinase, STY46, in Regulating the Sugar Starvation Response in Arabidopsis thaliana

Student Name: Marlene Goetz
UCD Mentor: Dr. Diane Beckles

When plants undergo environmental stresses, they experience sugar starvation (SS), causing a shift in carbon fluxes and signal transduction cascades to alter the plant’s metabolism and growth processes. This series of events, called the sugar starvation response (SSR), is crucial in determining plants’ survival under diminished carbon levels. It is speculated that the protein kinase, STY46, is a potential master regulator of the sugar starvation response. Compared to wild type plants, Arabidopsis with genetically altered levels of STY46 are expected to have differences in carbon use and plant growth. To meet this objective, four transgenic Arabidopsis thaliana lines (knockout, complementation, inducible overexpression and constitutive overexpression) will be generated to examine carbon partitioning and allocation under sugar starvation. The specific aim of this paper was to generate construct for a STY46 complementation line and discuss future research to be performed using the constructs. Positive results were obtained in preliminary gel electrophoresis, since restriction enzyme digest of the transformed plasmid construct appeared accurate. However, when the STY46 gene was sequenced, the results were inconclusive due to the inability of the primers to bind onto the plasmid. This most likely occurred due to either incorrect primer design or untransformed plasmids sent to sequencing. 

Biomarkers to Signify Early Detection of a Spontaneous Mouse Mutation In the Palmitoyl Transferase Zdhhc13 Gene

Student Name: Matthew Xie
UCD Department: VM: Molecular Biosciences
UCD Mentor: Dr. Cecilia Giulivi

The Zdhhc13 gene in mice is partly responsible for the encoding of palmitoyl acyltransferase (PAT) enzymes in mice. A naturally occurring recessive mutation in the mice causes a nonsense base substitution, leading to a truncated form of the Zdhhc13 protein. Mutant mice experienced an increased susceptibility to tumor multiplicity and malignant progression of papillomas after chemically induced skin carcinogenesis. Our goal in this research is to identify potential biomarkers that signify early detection of mutation in the Zdhhc13 gene. This is mainly done through MetaboAnalyst, a metabolite analysis program used to identify possible biomarkers, achieve group separation, and analyze pathways of metabolites to see their role in a functional gene. After the metabolite concentrations from three groups of mice are normalized, statistical procedures are conducted, and significant compounds are identified. Our graphs showed fair separation between homozygous WT, heterozygous, and homozygous mutant mice, with heterozygous mice showing characteristics closer to homozygous mutant mice than homozygous WT mice. Significant biomarkers identified include cysteine, taurine, L-pyroglutamic acid, and L-glutamic acid, which all have a direct role or a connection to processes in the skin.

Kinetic Analysis of Three Glycoside Hydrolase Mutants to Discover Associations Between Michaelis-Menten Constants and Protein Structure

Student Name: Michael Tan
UCD Department: Med: Biochemistry & Molecular Medicine
UCD Mentor: Dr. Justin Siegel

The need to process and analyze novel enzymes is exponentially growing; meanwhile the rate to do so has not kept up. Therefore, computational methods are necessary to continue to explore an ever increasing amount of new enzymes. This project intends to enhance computational methods through the collection of experimental data from mutant proteins to further determine how protein structure affects enzyme efficiency. To do so, the Michaelis-Menten kinetic constants, Kcat and KM values, will be gathered on three different glycoside hydrolase mutants; subsequently, the data will be contributed to an algorithm that will better predictions of kinetic constants based on protein structure through computational modeling.

Understanding the molecular basis of Postharvest Chilling Injury in tomato fruit: Generation of RD29A::CBF1 transgenic construct 

Student Name: Michelle Jiang
UCD Mentor: Dr. Diane Beckles

For decades, consumers have been purchasing refrigerated produce and storing it in refrigerators at home. Although this approach helps to preserve the produce for a certain period of time after harvest, many fruits and vegetables are damaged by the cold in a disorder called Postharvest Chilling Injury (PCI). Although PCI has been extensively studies, the molecular basis of its early events still remain poorly understood. The general goal of this project was to study those processes in tomatoes. The C-binding factor 1, or CBF1 gene, regulates cold tolerance, and the objective is to introduce CBF1 from a cold tolerant species, Solanum habrochaites, into the cold-sensitive cultivated tomato, Solanum lycopersicum. The aim of this work was to synthesize RD29A::CBF1 gene constructs which will be transformed into S. lycopersicum to assess cold tolerance in fruit after harvest. The two constructs created were RD29A::ShCBF1 using CBF1 from S. habrochaites and RD29A::SlCBF1 using CBF1 from S. lycopersicum. RD29A, ShCBF1, and SlCBF1 were amplified using Assembly Polymerase Chain Reaction (PCR). After purification, RD29A::ShCBF1 and RD29A::SlCBF1 constructs were spliced into pCAMBIA1300 vector plasmids and transformed into competent E. coli cells which copied the plasmids after multiple rounds of replication. The plasmids were extracted and diagnostic restriction digest was conducted with the EcoRI enzyme which yielded positives in colony #1 of RD29A::ShCBF1 and colony #21 of RD29A::SlCBF1. These were then sent for sequencing. This portion of the project culminated without the corroboration of the identities of the two engineered gene constructs due to time constraints. Even so, this brings research one step closer to reducing PCI in produce and decreasing the overall losses in agriculture.

Computational Design of Single Point Mutations on the Beta-Glucosidase B (BglB) enzyme

Student Name: Michelle Zhou
UCD Department: Med: Biochemistry & Molecular Medicine
UCD Mentor: Dr. Justin Siegel

Enzymes play a fundamental role in catalytic functions integral to many biological processes. The use of computational design to engineer the structure and predict enzymatic function hold the potential to enhance catalytic efficiency. Here, 3-D protein-folding simulators were used to structurally mutate the BglB enzyme’s primary structure for three mutants. The computationally designed mutants, then produced and purified, were analyzed according to Michaelis-Menten kinetics that provided further insight on the mutants’ catalytic efficiency (Kcat/Km). The compiled data will be important for developing and refining computational algorithms that can successfully predict correlations between structure and function. 

Observing the Different Stress Tolerances of Lactobacillus plantarum

Student Name: Nathan Lee
UCD Department: Food Science and Technology
UCD Mentor: Dr. Maria Marco

In today’s fermented foods industry, a single harmful type of bacteria can destroy a whole crop yield. However, the presence of lactic acid bacteria can kill these detrimental types of bacteria because of its acidic nature. Nathan’s experiment focuses on testing the different stressful environments LAB can endure so as to give more concrete data on the environments in which LAB can grow and be of service to farmers and agricultural companies.

The Effects of Restrictive Cardiomyopathy R145W Mutation on Signaling Pathways and Proteasome Function

Student Name: Rebecca Chang
UCD Mentor: Dr. Aldrin Gomes

The leading cause of sudden cardiac death (SCD) in young adults is familial hypertrophic cardiomyopathy (FHC), but the incidence of SCD in restrictive cardiomyopathy is even higher. Restrictive cardiomyopathy is a heart condition where the ventricle walls do not thicken but become stiff and cannot relax. Although restrictive cardiomyopathy is the least common of the cardiomyopathies, it has the highest mortality rate. However, how restrictive cardiomyopathy affects the heart is not well understood. The goal of this study was to determine the effects of the restrictive cardiomyopathy R145W mutation on signaling pathways and proteasome function in the heart. In the experiment, hearts with the mutation as well as control wild type hearts were homogenized in urea, centrifuged, digested with trypsin, and tagged with TMT isobaric mass tags for mass spectrometry. Statistical analysis showed that six proteins from the Parkinson’s pathway were present in the R145W hearts in significantly lower levels than in the wild type hearts. These proteins were proteasome subunits and heat shock proteins and were responsible for protein degradation and stress response in the cell. The lower amounts of these proteins suggests that the mutated hearts were likely subjected to higher amounts of oxidative stress due to the lack of response from the heat shock proteins and lack of proteasome activity that would normally help to prevent stress. With this research suggesting that R145W hearts are likely to experience more stress, future tests can be done to detect levels of oxidative stress, reactive oxidant species, ER stress related proteins, and determine the antioxidant status of animals.

Comparing the Targeting of Toc75 and OEP80, two Transmembrane Chloroplast Proteins, Through the Construction and Expression of a Chimeric Gene Involving Portions of Toc75 and OEP80.

Student Name: Renee Radusewicz
UCD Department: Plant Science
UCD Mentor: Dr. Kentaro Inoue

According to the endosymbiotic theory, chloroplasts descend from a cyanobacterial ancestor that was engulfed by a eukaryotic cell. Within the chloroplast’s outer membrane are paralogous transmembrane proteins known as Toc75 and OEP80. The two proteins evolved from a cyanobacterial protein, which inserted proteins into the outer membrane of the chloroplast’s cyanobacterial ancestor. Toc75 has evolved to function in protein import into the endosymbiont, which was required for the evolution of chloroplasts. The molecular function of OEP80 remains unknown, but is hypothesized to retain the function of its cyanobacterial ancestor. We are interested in how Toc75 evolved its novel function. One aspect of Toc75 and OEP80 that may contribute to their different functions is how they are targeted to the outer membrane of the chloroplast. Much is known about the targeting mechanism of Toc75, but not that of OEP80. Preliminary results of the Inoue lab indicate that Toc75 and OEP80 have distinct targeting mechanisms. To compare the targeting mechanism of OEP80 and Toc75, polymerase chain reactions (PCR) and gel electrophoresis will be utilized to construct a gene that incorporates portions of Toc75 and OEP80. The section in Toc75 that is responsible for targeting will replace the corresponding portion of OEP80. After Gateway cloning, plasmid isolation, transformation of agrobacteria, and infiltration of the bacteria into Nicotiana benthamiana, this transgene will be expressed in plants. The chloroplasts of the mutant plant cells will then be isolated and undergo SDS-PAGE and a Western blot, enabling this project to verify where the protein produced by the new gene is located within the chloroplast. This will determine if difference in targeting mechanisms between the two transport proteins contributed to distinct functions, hinting at how chloroplasts evolved through endosymbiosis. 

Computational investigation of peculiarly-conserved elements between the C. elegans and C. briggsae genomes

Student Name: Richard Yu
UCD Department: MCB
UCD Mentor: Dr. Ian Korf

We define peculiarly-conserved elements (PCEs) as regions of DNA at least 100nt long and conserved at unusually high levels between multiple organisms. In the genomes of the C. elegans and C. briggsae nematodes, we found a total of 385 PCEs. We computationally investigated and analyzed these PCEs and found that most of them are protein-coding, lie on genes, lie on exons or intron-exon junctions, code for mRNA, and have richer G+C content than DNA of typical conservation. All intergenic PCEs were found to lie on the X chromosome. These unique properties may facilitate identification of unusually high DNA conservation in nematodes. Furthermore, these findings introduce new areas of study in nematode, organismal, and evolutionary biology.

Potential of Tyrosine Sulfation in Voltage-Gated Sodium Channels

Student Name: Sabrina Liu
UCD Mentor: Dr. Grace Rosenquist

Tyrosine sulfation is known to be a key process in many organisms but the lack of an efficient way to figure out which tyrosines are sulfated means that the sites need to be predicted. Voltage-gated sodium channels have many potential tyrosine sulfation sites, especially GRNPNYGYTSF, MAMEHYPMTDH, and YEESLYMYLYF, that have evidence based on homology, mutagenesis, and toxin binding that support their sulfation. Their connection to various muscle and nervous system conditions can help the development of new drugs. However, more research will be needed to confirm sulfation in these sites.

Canopy and leaf temperature, stomatal conductance, and stem water potential in greenhouse tomato plants sensitive to water stress

Student Name: Sarah Hancock
UCD Mentor: Dr. Kenneth Shackel

Examines how greenhouse tomato plants respond to water stress by measuring different physiological responses of the plants, including leaf and canopy temperature, evapotranspiration, stomatal conductance, and water potential. The long term goal of this research is the development of a sensor that manages irrigation for field crops based on their water needs. While there currently exists many methods for gauging plant water stress, none of them are particularly accessible, especially in the context of the large-scale agriculture of modern society. This experiment is designed to affirm the link between canopy temperature and water stress by comparing canopy temperature trends to trends measured with conventional techniques, so that canopy temperature can be used to detect water stress across a large agricultural area.

Modifications to Brush Border Enzyme Glycosylation Decreases Enzymatic Activity

Student Name: Sithara Menon
UCD Department: Vet Med
UCD Mentor: Dr. Helen Raybould

Glycosylation is an important modification to cell surface proteins. The brush border of intestinal epithelial cells (IECs) contains many glycoproteins that are key to intestinal function. This study tested how changes to glycosylation affects the function of cell surface glycoproteins in IECs. Specifically, how removing N-glycans with exoglycosidases, such as those secreted by intestinal bacteria, affects the function of brush border enzymes, such as Intestinal Alkaline Phosphatase and Dipeptidyl Peptidase IV. This was tested in Caco-2 cells, a human cell line derived from colon adenocarcinomas that is used as a model for intestinal function. Caco-2 cells were cultured for 12 days, and their expression of IAP was tracked throughout the differentiation process. Once they were fully differentiated, the cells were then treated with glycosidases, and the enzymatic activity of IAP and DPP IV was measured. The results showed that changing the glycosylation caused enzymatic activity to decrease. Removing all the N-glycans with the glycosidase PNgase F decreased IAP activity to about 40 %, confirming that glycans are key to the function of the enzyme. Of the individual monosaccharides, removing mannose had the greatest effect, and removing sialic acid and fucose also affected the enzymatic activity. However, removing individual sugars did not cause any large decreases in activity, especially compared to removing the entire glycan.

A Mechanistic Insight on Chemical Enhancement of Gold Nanoparticles through Electron Paramagnetic Resonance

Student Name: Sonny Huynh
UCD Department: Chemistry
UCD Mentor: Dr. Ting Guo

Nanoparticles are capable of enhancing reactions under X-ray irradiation. In order to utilize this enhancement and apply it to other reactions, further knowledge of the enhancement mechanism is required. The experiment was performed in two parts, the first involving the quantification of chemical enhancement using a fluorometer to test the fluorescence of the hydroxylation of coumarin carboxylic acid in the presence of various concentrations of gold nanoparticles. The second part of the experiment involved understanding the hydroxyl radical, reactive oxygen species (ROS), pathway of the reaction using electron paramagnetic resonance (EPR). It was found that the surface of gold nanoparticles catalytically facilitates conversion 3-OH radical adduct to 7-OH-CCA in the presence of oxygen. Also the amount of hydroxyl radicals found in solutions with varying concentrations of gold nanoparticles was constant, implying that ROS generation is not a part of the catalytic process. 

Detecting Mutations and Heteroplasmy in Mitochondrial DNA with Surveyor Endonuclease.

Student Name: Sung Bin (Sean) Roh
UCD Department: VM: Molecular Biosciences
UCD Mentor: Dr. Cecilia Giuvili

Due to the susceptibility of mitochondrial DNA to insertions, deletions, and point mutations and the existence of multiple copies of mitochondrial DNA in each organelle, a phenomenon called heteroplasmy arises. High levels of heteroplasmy are known to be a factor in the development of mitochondrial diseases. Sung Bin “Sean” is optimizing a procedure to detect low levels of heteroplasmy. Applications of this research include future studies into heteroplasmy as well as diagnosis of risk of mitochondrial diseases.

The Effect of Computationally Designed Point Mutations on Structure and Kinetic Characteristics of Glycosidase Hydrolase Enzymes

Student Name: Willy Fan
UCD Department: Med: Biochemistry & Molecular Medicine
UCD Mentor: Dr. Justin Siegel

The importance of predicting an enzyme’s function and rate of function based on its structure. Due to the inaccuracy of current predictive algorithms, there is a need for large and comprehensive databases from which more accurate predictions can be made. Three mutants of the beta glucosidase B enzyme were tested for expression and then kinetically characterized through Michaelis-Menten kinetics. The ultimate goal of this project is to be able accurately mutate an enzyme for a specific purpose (i.e increased or slowed activity) and even create novel enzymes from scratch with the intended chemical activity.

Salinity stress, a key factor affecting almond tree stem water potential

Student Name: Zhiren (George) Ye
UCD Department: Plant Science
UCD Mentor: Dr. Patrick Brown

Almond trees are particularly sensitive to salt; their perennial nature allows the damage to the tree to cumulate over time. An estimated 10% of almond acreage is salt impacted and an additional 30% is at immediate risk. Research was conducted to investigate how NaCl applications affect the plant water status of almond trees. It was hypothesized that addition of salts would decrease stem water potential over time. The experiment consisted of two trees: one under balanced nutritional conditions and the other under salinity stress. Five leaves were collected from each tree around noon daily and stem water potential of each leaf is measured using a pressure chamber. Data collected showed that there was no significant decrease of stem water potential for the first 10 days. However, stem water potential values after 10 days showed a decreasing trend. Results were consistent with the hypothesis after 10 days of NaCl applications. Addition of salts indeed decreases stem water potential over time. Different salt concentration may be applied to evaluate the change in almond tree growth and yield in the future. Ion toxicity of almond trees may also be investigated by comparing the effect of polyethylene glycol and sodium chloride.

2015
 

Structure-Seq Influenced RNA Secondary Structure Prediction

Student Name: Aditya Bollam
UCD Department: Biomedical Engineering
UCD Mentor: Sharon AViran

Current life on earth is believed to have descended from self-replicating RNA molecules, and evidence of its deep history is apparent in its diversity of function. Discerning the structure of RNAs will help inform our understanding of RNA function as well as develop future tools in medicine. The most basic structural information lies in its secondary structure, the first level of structural organization within an RNA molecule. Techniques such as crystallography, comparative analysis, and computational algorithms have been developed to predict secondary structure of RNA, though performance becomes hindered when analyzing longer RNAs. In order to find a more efficient method of predicting the structure of long RNAs, we combine data gathered from DMS probing experiments (Structure-Seq) and input subsections of the RNA into the RNAStructure prediction algorithm. By dividing the structure into smaller sections, we find that predictive capabilities can be vastly improved, though inclusion of DMS probing data has varying effects in improving prediction accuracy. We test this subdividing of RNA of prediction in both a user-directed and naive manner in the 18S RNA in Arabidopsis thaliana. Overall, these improvements in computation and experimentation suggest a more efficient and accurate strategy to predict RNA secondary structure in long RNAs.

Biochemical Characterization of MAPK6 Enzyme in Regards to Plant Innate Immunity Response Pathways

Student Name: Alex D. Doan
UCD Department: Department of Plant Biology and The Genome Center
UCD Mentor: S.P Dinesh-Kumar

Plants and animals have evolved and developed different methods of immunity to defend themselves against various attacking pathogens. One technique is by recognizing such pathogens through recognition receptor proteins located on the extracellular membrane and then signaling a response pathway. In this lab, the MAPK6 enzyme of the MAP/ERK pathway in Arabidopsis thaliana was studied and biochemically characterized. The gene for the protein was cloned, expressed in bacteria and the protein then purified through extraction. By running an analytical gel exclusion chromatography, MAPK6 was then determined to be monomeric structurally. Also, through an in vitro reaction with ATP, the protein was determined to be an active kinase that utilizes phosphorylation to transmit immune responses within the cascading pathway.

A Plant-based Evaluation of Landscape Irrigation Efficiency

Student Name: Brian Glucksman
UCD Department: Department of Pomology
UCD Mentor: Ken Schackel and Maya Lopez-Ishikawa

To accommodate a rapidly drier climate, Californians need to cut water from landscaping. The goal of this study is to evaluate the effectiveness of adding extra water to young frontier elms (Ulmus “Frontier), using hydration bags on young valley oaks, and irrigating young valley oaks with MP rotators and hydration bags. The plant-based measurement of water stress, midday stem water potential (MSWP), was collected in the early afternoon through the process of pressure bombing. This study found that adding extra water did not reduce the water stress of frontier elms and had no prolonged effect, that using hydration bags was three times as water efficient as using MP Rotators at watering the young valley oaks, and that removing hydration bags increased water stress in young valley oaks. 

Arabidopsis Thaliana Cell Plate Formation: an Interdisciplinary Approach to Characterizing ES7 Resistant Mutants

Student Name: Emily Dickinson
UCD Department: Plant Biology
UCD Mentor: Dr. Georgia Drakakaki and Destiny Davis

There is currently a lack of knowledge surrounding the signals and factors that lead to cell plate formation during cytokinesis. Although scientists in the field know the multistep process of the cell plate formation, there is still much to be learned about the molecular events occurring during formation. The Drakakaki lab at UC Davis is studying endomembrane trafficking and polysaccharide deposition at the cell plate. By using the chemical Endosidin 7(ES7), which inhibits cell plate maturation by specifically impeding callose deposition, the researchers have developed mutant strains of the Arabidopsis thaliana that are resistant (Park et. al 2013). Characterization of two mutants es7r-1 (ems 13) and es7r-3 (ems 70) by root length and subcellular effect in response to ES7 will yield the target gene and therefore a novel protein active during cell plate formation. In addition, a cell plate vesicle proteomics approach will lead to a collection of cell plate-destined proteins which to study further

Cloning and Preliminary Characterization of STY46 from Arabidopsis thaliana, a Candidate Protein Kinase Involved in the Plant Sugar Starvation Response

Student Name: Haishan Gao
UCD Department: Department of Plant Sciences
UCD Mentor: Diane M. Beckles

To study how plants respond to conditions under which the carbohydrates needed for growth and development are exhausted, this project focuses on cloning and functional characterization of a cytosolic protein kinase, STY46 that may potentially be involved in the sugar starvation response (SSR). Generating A. thaliana STY46 knockout (-KO) and overexpression (-OE) lines is the main method in this project to determine the function of the protein. A SALK line (SALK_116340) was confirmed to contain a T-DNA insert in the STY46 gene and the aim was to prove that this reduced the expression of this gene. Highly intact RNA was isolated and reverse transcription PCR performed to determine the level of transcript of STY46 in STY46-KO. However the results were inconclusive. This may be due to not enough amount of RNA use for the cDNA reverse transcription. To develop an overexpression line, STY46 was cloned from wild type Arabidopsis genotype. Primers were designed to flank the open read frame of the STY46. They also contained restriction enzyme recognition sites and a myc-tag engineered at their 5’ ends to facilitate subsequent synthesis of a plant transformation construct. A cDNA fragment of the expected size was successfully amplified by PCR. 

Role of Mitochondrial Inner Membrane Complex Organizing System in Fibroblasts from FMR1 Premutation Carriers

Student Name: Jacob Eisner
UCD Department: Molecular Biosciences: Vet Med
UCD Mentor: Dr. Cecilia Giulivi and Sarah Wong

FXTAS is an adult onset neurodegenerative disorder known to cause symptoms such as intention tremor and gait ataxia, which grow in severity with age. It is also possible to have the FMR1 premutation that causes FXTAS without experiencing symptoms of the disorder. In this study, fibroblast samples isolated from FMR1 premutation carriers with and without FXTAS and age and sex matched controls were tested through PCR in order to determine the gene expression of MICOS protein encoding genes and the FMR1 gene. Samples from male premutation carriers displayed a downward trend in expression of MICOS protein encoding genes and the FMR1 gene as age increased, whereas samples from female premutation carriers exhibited the opposite trend. Although this pilot study does not provide enough conclusive data to accurately and precisely justify any conclusions, it can be inferred from these data that males overexpress the genes tested while young in order to compensate for deficits associated with the premutation and resist pathogenesis until they grow too old to do so effectively, whereas FMR1 premutation heterozygous females underexpress the genes tested while young due to a compensatory response in the remaining unaffected allele.

Quick Screening of Chemoreceptor Functions

Student Name: Jimmy Kim
UCD Department: Department of Microbiology and Molecular Genetics
UCD Mentor: Dr. Rebecca Parales and Watumesa Tan

Chemotaxis allows some organisms like Pseudomonas bacteria to move in response to chemical stimuli. Methyl-accepting chemotaxis proteins (MCPs) are membrane-bound chemoreceptors that facilitate this process. Understanding MCPs could lead to potential application of chemotaxis in fields like bioremediation, where it could improve efficiency since bacteria could not only break down pollutants but seek them out as well. In order to characterize the MCPs of species like Pseudomonas putida F1, hybrid sensor proteins comprised of the sensory domain of bacterial MCPs and the signaling domain of a 2-component regulator were used in the optimization of a screening method. By using a filter paper disk to diffuse potential attractant/repellants in a plate, bacteria with these hybrid sensor proteins will show an inhibition of growth if they sense the compound. This screening method was optimized by testing various concentrations and volumes of compounds as well as plate media content. This method will allow for quick screening of MCP functions in P. putida F1 and other bacteria.

Buffering Capacity of Foods and its Effects on Digestion

Student Name: Jonathan Liu
UCD Department: Food Science and Technology
UCD Mentor: Dr. Bornhorst, Dr. Ferrua, and Dr. Singh

The buffering capacity of foods plays a pivotal role in food breakdown during gastric digestion. The ability of foods to resist changes in pH alters the acidity of the stomach, influencing acid secretion, enzyme function, and chyme viscosity[SO1] . All of these properties directly influence the stomach’s ability to break down foods. Therefore, a comprehensive and replicable method to determine the buffering capacity of foods would prove beneficial for helping to predict and compare digestive properties [SO2] of different foods. We have developed a preliminary methodology for determining the buffering capacities of certain categories and subcategories of undigested foods, and used these to determine buffering properties of certain semisolid foods before digestion.

Cocrystallization of C60 and C70 Mixture with Sulfur

Student Name: Karen C. Guo
UCD Department: Department of Chemistry
UCD Mentor: Alan L. Balch

Research and application of fullerenes have been limited by the work and cost of separation and purification. This paper studies sulfur as a separation agent with C60 and C70 solutions. Crystal tubes were made of sulfur and 1:1 molar ratio mix of C60 and C70. Resulting crystals were tested with HPLC and single crystal X-Ray diffraction. It was found that sulfur cocrystallizes with both C60 and C70, but the two fullerenes crystallize separately. The results of this experiment can be used to develop new separation methods for fullerenes and adds to existing knowledge of sulfur as a cocrystallizing agent with fullerenes. 

Determining the Role of OEP80tr in Plant Germination and Growth

Student Name: Katie Chung
UCD Department: Plant Biology
UCD Mentor: Kentaro Inoue and Philip Day

The outer membrane protein 85 (Omp85) family proteins found in the outer envelope of chloroplasts are remnants of ancestral cyanobacterium protein since their endosymbiosis by eukaryotes.  Flowering plants in particular have an additional beta barrel protein in the outer membrane of their chloroplasts, OEP80tr, whose exact function is unknown.  This study attempts to determine the function of OEP80tr by investigating the effects of null expression mutations of the gene.  Through analysis of germination and plant growth (in roots, hypochotyl, and leaves), this study compares the phenotypes of wild type plants with the functioning gene to plants without the gene.  The resulting observations could then be used to determine the role in development of the OEP80tr protein. 

Reconciling Cost Efficiency and Biodiversity by Native Seed Dispersal

Student Name: Lauren Lee
UCD Department: Plant Sciences
UCD Mentor: Aubrianne Zamora and Emilio Laca

Restoration of native grasslands in California is expensive, mostly due to the high cost of seeds. Although restoration can increase biodiversity and ecosystem services, ecosystem services do not generate revenue. In order to reconcile economics and the need for biodiversity in California annual grasslands, this study focuses on discovering how a grass species’ status as an annual or perennial plant affects biodiversity and relates to cost. We studied several restoration methods based on strip seeding, where strips of various widths were alternatively seeded or left unseeded. We hypothesized that over time seeds from the seeded strips would establish in the unseeded strips, a cost-efficient means of restoration. Requiring $825 and $1250 respectively, the 33% seeded and 50% seeded (wide) treatment demonstrated potential as cost efficient means of habitat restoration. The 33% treatment resulted in approximately 12.5 species at the plot level for species richness, while the 50% wide treatment yielded 12.4 species. Botanical composition of seeded and unseeded strips indicated that strip seeding is a successful low-cost method for Elymus glaucus, but other native perennial grasses failed to establish in unseeded strips. 

Understanding the Structural and Molecular Basis of Postharvest Chilling Injury in Tomato Fruit (Solanum lycopersicum L.)

Student Name: Leona Lee
UCD Department: Plant Science and Food Science
UCD Mentor: Diane M Beckles and Michael McCarthy

Postharvest Chilling Injury (PCI) is a devastating disorder that leads to loss of produce. In this study two approaches were taken to better understand the development of PCI in tomato (Solanum lycopersicum L.) a species susceptible to PCI. First, the spatio-temporal development of PCI symptoms in developing fruit was assessed using Magnetic Resonance Imaging (MRI). Second, the CBF1 gene from Arabidopsis thaliana was cloned so that the gene may be engineered into tomato fruit to improve the tomato fruit response to chilling. MRI data indicates that chilling injury affects each tomato tissue type (the pericarp, the locular tissue and the columella) differently with greater effects seen in green fruit due to their continuation of ripening and various metabolic processes. A 780 bp fragment corresponding to the coding sequence (CDS) of CBF1 was amplified from Arabidopsis genomic DNA and attempts to sub-clone into pGEM-T Easy Vector® are underway.

Optimization of Homogenization Protocol for Analysis of TPPII in Heart Samples

Student Name: Meera B. Ganesh
UCD Department: Department of Neurobiology, Physiology and Behavior
UCD Mentor: Dr. Aldrin V. Gomes

The primary function of proteasomes is to degrade mis-folded and unwanted proteins. Tripeptidyl peptidase II (TPPII) is a newly discovered protease that works downstream of the ubiquitin-proteasome system. It is required in eukaryotic cells for cleaving of larger protein degradation products. However, little is known about its function in other cellular processes. TPPII’s responses to stress and DNA damage could be areas of research for cancer therapy, if TPPII can be studied better.  Homogenization is a process used to break down cells for subsequent analysis or purification of proteins. Accurate analysis of protein expression and protease activity in the cell can be studied by good homogenization procedures and sample preparation. This study investigated the effect of various homogenization and assay buffers to determine the optimal buffer for homogenization and activity assays of cardiac TPPII. The results demonstrated that a homogenization buffer containing polyvinylpyrrolidone and glycerol produced more desirable results compared to buffers currently used for TPPII analysis

Structural and kinetic analysis of hydrolase mutants and subsequent impact on predicting functionality with Michaelis-Menten mechanics

Student Name: Michael Hsiu
UCD Department: Department of Chemistry
UCD Mentor: Justin Siegel and Ryan Caster

Understanding about enzymes has grown due to the use of computational algorithms in predicting the correlation between structure and function. These algorithms have had critical roles in the redesign and reengineering of native enzymes to enhance catalytic behavior. In this paper, front-end programs were used to model, interact with, and redesign a glycoside hydrolase enzyme. The structural model was coupled with Michaelis-Menten kinetics to yield the Kcat and Km kinetic constants for three mutants. The compiled dataset will be essential to tracking patterns and trends across enzyme families. It may also be useful in future efforts to redesign enzymes using predictive methods. 

The Effect of Stem Heliotropism on Sunflower Growth and Productivity

Student Name: Nicole Infantino
UCD Department: Plant Biology
UCD Mentor: Dr. Stacey L. Harmer and Dr. Hagop Atamian

Heliotropism, also known as solar-tracking, is the diurnal motion of plant leaves and flowers in response to the direction of the sun.  Many studies of solar-tracking plants indicate that heliotropism is correlated with higher light absorption, photosynthetic efficiency, and crop productivity.  Sunflowers undergo this process by orienting their leaves, as well as their stem toward the sun throughout the day.  The flowers continue to move at night in order to readjust their direction in preparation of the sun’s rise in the east the next day.  This paper reports two experimental tests of the hypothesis that disrupting the normal solar cycle of the sunflower will slow the rate at which the plant develops and decrease yield.  In the first experiment, 83 HA412 inbred line sunflowers were planted in a field.  Half of the plants were treated by being tethered to bamboo stick in order to restrain heliotropic stem movement.  In the second experiment, 28 HA412 inbred line sunflowers were planted in 3 gallon pots placed in the field.  Half of the plants were rotated 180 degrees at sundown to face the other direction.  Leaf area, leaf count, and plant height was measured for all plants in both experiments.  The angle of head movement was also measured for the plants in the second experiment.  The results of both experiments show that plant growth and development was less in treated plants then the control plants, indicating that heliotropism plays a part in plant growth.  However, more research and trials are needed to confirm this.  The objective of this study is to determine the effect of stem heliotropism on sunflower growth and productivity in order to arbitrate whether genetically modifying other crop-bearing plants to elicit stem heliotropism would be beneficial to crop yield.

Prediction of Tyrosine Sulfation in Voltage-Gated Potassium Channels

Student Name: Savitri Asokan
UCD Department: Department of Neurobiology, Physiology and Behavior
UCD Mentor: Grace L. Rosenquist

Tyrosine sulfation is a protein modification which causes a tyrosine residue to gain a sulfate group and become negatively charged.  So far, 102 tyrosines have been shown to be sulfated, but no voltage-gated potassium channel proteins contain confirmed sulfated sites.  Using a Position-Specific-Scoring-Matrix, the selectivity filter GYGD and pore helix site MYP were predicted to likely be sulfated. Sequence alignment of these motifs across 40 potassium channel proteins showed that both motifs are conserved to a high degree, indicating some likely functional role.  Past mutagenesis experiments on the two sites were collected to determine the effect of the GYGD and MYP residues on toxin binding.  GYGD and MYP appear to be critical sites for cation binding, but no evidence of a strictly ion bond is available.

The Effects of Anaerobic Soil Disinfestation Treatments With Different Carbon Sources on Agrobacterium tumefaciens

Student Name: Shanti Gurbuxani
UCD Department: Plant Pathology Department
UCD Mentor: Daniel Kluepfel

As the worldwide restrictions for soil chemical fumigants increase, growers need alternatives to eradicate soilborne plant pathogens. Anaerobic soil disinfestation (ASD), developed in Japan and the Netherlands in the 2000s, is one of these alternative techniques that can control certain soilborne plant pathogens, plant-parasitic nematodes, and weeds in specialty crop systems. To utilize the technique, the soil has to have a carbon source applied, covered with plastic, and mixed with water to generate anaerobic conditions in the soil. The point of this experiment was to see the effects of different carbon sources such as ethanol, almond hulls, almond shells, and rice bran on the soilborne plant pathogen Agrobacterium tumefaciens, which causes crown gall disease in walnuts. The results showed that ethanol was the most effective ASD treatment at suppressing A. tumefaciens. However, the most anaerobic treatment was the almond shells, but it was also the treatment with the greatest abundance of A. tumefaciens. The anaerobic environment created by ASD may not be the only driving mechanism behind suppression of A. tumefaciens. The ethanol treatment did not have anaerobic conditions, but volatile compounds produced through ASD in the ethanol treatment could have suppressed the A. tumefaciens instead.

Determining the Signaling Pathways Regulated by Chaperone-Mediated Autophagy

Student Name: Sriya Lingampalli
UCD Department: Department of Neurobiology, Physiology, and Behavior
UCD Mentor: Aldrin V. Gomes

Chaperone-mediated autophagy (CMA) is a process that maintains cellular homeostasis and degrades dysfunctional or damaged proteins. It functions by transporting proteins to the lysosome for degradation with the help of the HSC70 and LAMP-2A molecules. Research demonstrating a five residue amino acid sequence and several substrate proteins has resulted in many proteins being identified as potentially CMA regulated. We analyzed all proteins in the genome of several species including humans, and concluded that many proteins and signaling pathways are likely CMA regulated. Using a novel prediction program, we have also found that a greater percentage of proteins (46.5%) in the human protein sequences possess KFERQ-like motifs than previously thought (30%). Testing of lysosomes from starved H9c2 cells showed that the three proteins, which are predicted to have KFERQ-like motifs, are present in the lysosome. 

Understanding The Role Of Salicylic Acid Signaling Pathway In Stromule Induction During Plant Innate Immunity

Student Name: Sydney Collier
UCD Department: Department of Plant Biology and the Genome CenterUCD Mentor: Eunsook Park, and Savithramma Dinesh-Kumar

      Stromules are plant cell specific structures that extend out from the chloroplast. The role of the stromule is not known, however, a recent publication from the Dinesh-Kumar lab suggests that they are involved in the plant’s immune response to protect the plant from pathogens. More detailed studies regarding the regulation mechanism about the stromule induction is crucial to understand their contribution in the plant immune response. The Dinesh-Kumar lab studies have suggested that the stromule induction pathway might be regulated through the plant hormone Salicylic Acid (SA) pathway that is essential to the plant immune response. Key genes in this pathway are SID2/EDS16 and ATG5. To obtain a better understanding of the role of SA in stromule induction, the sid2/eds16 and atg5 mutants were crossed to make sid2/eds16 atg5 double mutant. sid2/eds16 atg5 double mutant was identified by genotyping using polymerase chain reaction (PCR). The results show that there was a double mutant and we will introduce a stromule marker later to observe stromule induction. Characterization of stromule induction in single and double mutants will provide insights on how stromule induction is regulated during the plant immune response.

Investigating the Effect of Mutations on the Enzymatic Activity of Beta-glucosidase B

Student Name: Victor Lee
UCD Department: Department of Chemistry
UCD Mentor: Dr. Justin Siegel, Ryan Caster, and Alex Carlin

Beta-glucosidase B is an enzyme found in every organism that breaks down the glycosidic bondages of polysaccharide molecules. Currently, there is a lack of understanding of how enzymes work, partly because databases on how point mutations affect this enzyme are incomplete. In this study, we investigated the effects of how three specific point mutation affects the enzyme’s effectiveness by using kinetic constants to characterize the enzyme’s effectiveness. One mutation has significantly improved enzymatic activity, but the other mutations have caused the enzyme to be less effective. Investigations such as these could lead to an understanding of how enzymes work, which has many applications, such as the biofuels industry.

2014
 

A Comparison of Tripeptidyl-Peptidase II in Akita and common mice using western blotting

Student Name: Angie Chen
UCD Mentor: Dr. Aldrin Gomes

The activity and expression levels of tripeptidyl-peptidase II (TPP-II), a proteolytic protein complex, is unknown in diabetic muscle tissue. To investigate TPP-Iwestern blotting and biological assays were employed to answer the questions: 1) Do TPP-II levels differ in Akita mice versus wild-type mice? and 2) Is TPP-II activity different in Akita and wild-type mice? Before responding to these questions, however, the common western blotting technique used was optimized to improve the sensitivity of the blot. The optimal conditions for western blotting were determined to be 5% PEG-8000 + 0.01% Glutaraldehyde + 75% TTBS and overnight primary antibody incubation. Using this optimized western blotting, TPP-II levels were determined to be the same in Akita and wild-type mice. TPP-II activity was also determined and found to be similar between hearts from Akita and wild-type mice.

Understanding the Influence of Multiple Contextual Features During the Encoding of Episodic Memory

Student Name: Bethany Hung
UCD Mentor: Dr. Arne Ekstrom

The ability to form complex memories is a well studied yet not well understood mechanism of the brain. Several studies have been performed on the creation of episodic memories from object representations, but none have examined the effect of multiple types of contextual cues and how they are prioritized during memory encoding. In this study, we examined the creation of episodic memories via the integration of multiple different elements. Sixty-four subjects, all of normal health, navigated a system of virtual reality rooms and answered questions about the room and music’s emotional effect during a study phase. During the testing phase, they once again navigated the hallway but answered questions that tested for recall of details in rooms that they entered. We then quantified this data by counting corrects versus false alarms and analyzed the data using a one-way ANOVA. We examined three model effects: delay between the study and test phases, the subjects’ type of recall – spatial, musical, or both – as determined by the questions, and the interaction between delay and recall type. Our findings indicate that music, as a lone contextual cue, provides an advantage over spatial cues during encoding that later presented itself in retrieval. However, this effect dissipated after one week. It therefore appears that the impact of varying types of contextual details changes differently over time, although a combination of multiple details is always better than a single type of detail.

The effect of microwave intensity and exposure on the carotenoid concentration of a cassava gari meal with added red palm oil.

Student Name: Ethan Udell
UCD Mentor: Dr. Betty Burri

Cassava gari is a staple food in Africa, which has been biofortified to help combat vitamin A deficiency. However, a correlation is often seen between the extent of gari preparation and a decrease in the carotenoid concentration of the gari. The objective is to find out if higher microwave intensities and longer duration of exposure to microwaves may decrease the carotenoid concentration in cassava gari.

Spatio-temporal dynamics and physiological characterization of post harvest chilling in tomato fruit.

Student Name: Hansalia Sahil
UCD Mentor: Dr. Diane Beckles

Magnetic resonance imaging was used to spatially and temporally examine physiological changes in tomato fruit (S. lycopersicum var. cerasiforme cv. sweet 100) caused by exposure to chilling temperatures. Measurements for respiration and ethylene production showed the development of chilling injury as indicated by the chilling injury index. The MRI was used to calculate the apparent diffusion coefficient (ADC), a measure of water mobility in tissues, for different regions of interest (ROI) in the fruit.

Analyzing Lipid Structure on Oxidized Gold as a Function of Vesicle Size and Solution

Student Name: John Rodgers
UCD Mentor: Dr. Donald Land

Small unilamellar vesicles (SUVs) aptly model cell membranes.  In fact, vesicles are currently a major area of focus.  They are being used to study biological functions through the incorporation of various molecules into the vesicle membrane . In this research, the structure of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) 30 and 100 nanometer vesicles were studied in phosphate buffered saline (PBS) and water. The lipid samples were adsorbed on an oxidized gold surface on a germanium crystal and studied with attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR). In both PBS and water, 30 and 100 nm vesicles adsorbed intact. However, deformation of the SUVs varied with the size of the vesicles and solvent.  This study shows that DMPC vesicles reacted differently based on changes to these two factors.  These observations will help advance surface chemistry research and the development of better cell bilayer models.

Predicting Tyrosine Sulfation in GPCRs

Student Name: Kevin Ma
UCD Mentor: Dr. Grace Rosenquist

The posttranslational modification of tyrosine sulfation is critical for the binding affinity and peptide specificity of many G-protein coupled receptors (GPCRs). Using a log-odds position specific scoring matrix (PSSM), 64 tyrosine sulfation sites in 31 different peptide binding GPCRs were scored. Higher scores indicated a higher degree of similarity between confirmed sulfation sites and that tyrosine. While our PSSM did not account for characteristics such as clustering and conservation, the sites nonetheless exhibited these characteristics. Most predicted sites were located in the binding pockets of GPCRs, which is also consistent with confirmed tyrosine sulfation sites. Binding affinity or peptide specificity decreased when our tyrosines were mutated into alanine or phenylalanine, indicating the important role of the tyrosine in ligand binding.

The Role of Protein Ferritin in Plant Immune Responses to a Pathogen

Student Name: Lila Balakrishnan
UCD Mentor: Dr. Dinesh-Kumar

In N.tabacum, chloroplastic protein NRIP1 mediates the innate immune receptor recognition of p50, the viral effector protein of the Tobacco Mosaic Virus. The presence of stromules was observed  extending from the stroma of the chloroplasts towards the nucleus. Researchers discovered that ferritin, another chloroplastic protein, does not complete a similar process to NRIP1, instead, it is always located in small quantities in the nucleus that do not appear to change during N-mediated defense.

Comparing Glutathione S-Transferase Mu 1 in Akita Mice and Wild-Type Mice

Student Name: Madelyn Wang
UCD Mentor: Dr. Aldrin Gomes

The goal of this project was to determine if Glutathione S-Transferase Mu 1 (GSTM1), a key antioxidant enzyme, is altered in diabetes. We compared GSTM1 levels between hearts of type I diabetic Akita mice and wild-type mice using Western blotting. Western blotting for GSTM1 had previously been unsuccessful because of the low amounts of this enzyme in the heart. The Western blotting procedure was optimized through experiments to determine if treatment of the membrane prior to blocking and during primary and secondary antibody incubation increased the sensitivity of the blot. We found that treatment of the membrane in 0.01% glutaraldehyde for 20 minutes before blocking and incubating with the primary antibody in the presence of 75% TTBS (Tris-buffered saline with Tween 20) increased band intensity by 8.92-fold when compared to the control. In addition, the addition of 5% PEG-8000 and the use of 1% BSA in the primary and secondary antibody incubations increased Western blot sensitivity. The improved Western blotting procedure allowed for the detection and quantification of the GSTM1 in the Akita and control samples. After quantifying and normalizing the protein loaded, we found that GSTM1 levels were similar in Akita mice than in normal mice. Hence, the levels of GSTM1 do not seem to be affected in type 1 diabetic heart dysfunctions.

The Effects of Carbon Sources on Anaerobic Soil Disinfestation

Student Name: Matt Wang
UCD Mentor: Dr. Daniel Kluepfel

Greenhouse studies were performed to examine the effects of two carbon sources on Anaerobic Soil Disinfestation (ASD) of Agrobacterium tumefaciens and Pseudomonas synxantha inoculum. The first experiment was performed using Yolo silty clay loam soil from the UC Davis Armstrong Field Facility, which will be referred to as clay soil. The experimental design was composed of treatment pots in a controlled greenhouse environment, each with an irrigated soil amended with rice bran at a rate of 9, 7, or 5 tons/acre, or molasses at a rate of 4.5, 2.25, or 1.125 tons/acre. Inoculum bags containing the bacteria A. tumefaciens and P. synxantha were buried in these pots, which were then covered with an impermeable tarp. The second experiment only considered the treatment of 9 tons/acre of rice bran, but used soil with a Hanford fine sandy loam texture, which will be referred to as sandy soil. Untreated control pots were also present for comparison in both experiments. At three and seven days after initiating Anaerobic Soil Disinfestation, three pots from each treatment were taken to enumerate the bacteria through dilution plating on selective media. Populations of A. tumefaciens in the 1.125 tons/acre molasses treatment tended to decrease with time, but the populations at seven days were not significant different from time zero across treatments. Populations of P. synxantha in the 9 tons/acre rice bran treatment also tended to decrease with time, but the populations were also not significantly different from time zero across treatments. Anaerobic conditions were lost after five days for both the 1.125 tons/acre molasses 2.25 tons/acre molasses, with lower efficacy of ASD over time for molasses compared to the rice bran treatments. A. tumefaciens populations decreased to undetectable levels by seven days in the sandy soils exposed to ASD conditions. These data indicate that soil texture may play a major role in ASD efficacy which will affect where ASD will be used commercially in the state of California. Overall, the comparison of rice bran and molasses as carbon sources for ASD was inconclusive; however, it was clearly shown that rice bran at rates below commercial levels of 9 tons/acre were effective at inducing anaerobic conditions in the soil.

Refining the metagenomic approach used to study sanitizer-treated sludge microbiota and orange peels with limonene

Student Name: Ryan Poon
UCD Mentor: Dr. Christopher Simmons

To maximize biofuel methane production from food processor waste, our research sought to develop a system that could be used to genetically examine methanogens that are resistant to industrial and natural sanitizers. We especially focused on the orange juice processor waste, which contained the natural sanitizer limonene and the protein pectin that inhibited DNA extraction. Hermetic bioreactors filled with tomato pomace and sludge were connected to a respirometer, which quantified the gases they released. After testing factors such as the nutrient medium, it was found that higher volumes (>100 mL) of brand new sludge taken straight from refrigeration was needed for reliable results. The results of the following test trials showed that methanogens were increasingly hindered by increasing sanitizer concentrations above 30 mg/L, signifying that the system functioned properly. In another experiment, DNA was best extracted from a mixture of orange peels and limonene by lysing the samples without centrifuging or bead beating and using pectinase to digest the pectin. The concentration of DNA was qualitatively analyzed using spectrophotometry and Qubit®. For polymerase chain reaction (PCR) amplification, bovine serum albumin (BSA) was used to enhance the compatibility of the DNA strand as well as to prevent inhibitors from interfering with the process. Such a systematic procedure could not only be utilized to discover the genes coding for sanitizer resistance, but also potentially be applied to test a wide variety of variables that may affect methane generation.

2013
 

Towards Understanding the Importance and Divergence of the OMP85 homologs Toc75 and OEP80

Student Name: Aditya Srinivasan
UCD Department: Plant Sciences
UCD Mentor: Dr. Kentaro Inoue

This study focuses on the relationship between two OMP85 homologs, Toc75 and OEP80, derived from an ancestral cyanobacterial protein. These two homologs are present within modern-day chloroplasts. OEP80 is hypothesized to play an important role in seedling development much like its sister protein – Toc75. This study attempts to determine the function of OEP80 as well as its necessary expression time (in either the seed, germination, or seedling stage). By creating mutants without a properly functioning OEP80 gene and rescuing these mutants using genetic constructs, this study compares the functional sequences of OEP80 and Toc75. These constructs can then be used to determine the exact time (stage of plant development) and role that the OEP80 plays in functioning and normal development of plants. 

Prediction of Tyrosine Sulfated Sites in Integrins

Student Name: Allan Zhou
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Grace Rosenquist

Tyrosine sulfation is a post-translational modification of proteins that is important to protein-protein interactions. We show that integrins, a family of trans-membrane receptors found in animals that link cells to the Extracellular Matrix, have a high number of likely tyrosine sulfation sites. Particularly, likely tyrosine sites in integrins are often found near ligand binding or recognition sites. Our data also show a high degree of conservation between likely sulfated tyrosine sites, suggesting their functional importance to the integrin.

Effects of Dof1 Transcription Factors on Tomatoes

Student Name: Annabel Chem
UCD Department: Plant Sciences
UCD Mentor: Dr. Diane Beckles

Nitrogen is a nutrient essential to plant growth and development. Improving a plant’s ability to uptake and metabolize nitrogen is essential to preserving plant productivity and agricultural viability under low nitrogen growing conditions, but is difficult to achieve due to the complicated nature of the nitrogen assimilation pathway. Because transcription factors often regulates the coordinated expression of genes involved in a pathway, modification of transcription factors is a powerful and promising approach towards modifying complicated pathways. This experiment focuses on the effects that the Dof1 transcription factor has on nitrogen assimilation in tomato plants in hopes to design a tomato that can maintain high crop yield without requiring added nitrogen from fertilizers. Plant length, fresh weight, dry weight, root length, and PEP-Carboxylase expression were higher under low nitrogen conditions in plants overexpressing Dof1, showing that the Dof1’s role in regulating nitrogen metabolism in plants has a positive impact on nitrogen assimilation in transgenic tomato plants under low nitrogen conditions. Although this project evaluates the NUE of a transgenic tomato plant, the ultimate goal is to create a non-transgenic line that has similar properties. 

Roles of Arabidopsis thaliana Na+/H+ Antiporters NHX2 and NHX3 in Potassium Ion Homeostasis

Student Name: Athena Kan
UCD Department: Plant Sciences
UCD Mentor: Dr. Eduardo Blumwald

Antiporters are generally responsible for ion homeostasis in the model organism Arabidopsis thaliana by exchanging hydrogen ions (H+) for sodium (Na+) or potassium (K+). However, information regarding the role of NHX2 and NHX3 in this process is scarce; therefore, this study aims to investigate the effect of overexpression of NHX and NHX3 genes individually when the plant is grown in Spalding media with varying concentrations of potassium: 30mM (high), 1mM (medium), and 0.1mM (low). After ten days of growth, root length was measured and then expression of HAK5 and AKT1, which correlate with low levels and high levels of potassium, respectively, were measured using comparative qPCR analysis. The results indicated that though overexpression of the NHX2 gene had little to no effect in any concentration of potassium media, overexpression of the NHX3 gene significantly impacted growth and development. Specimens with an overexpressed NHX3 gene exhibited shorter roots, increased expression of HAK5, and decreased expression of AKT1 in low levels of potassium. Not only do these results present a better understanding of Arabidopsis thaliana, but they may also guide the process of developing better stress response capabilities in crops; abiotic stress is the primary cause of crop loss worldwide.

Effect of developmental stage on viability of medusahead seeds

Student Name: Catalina Zhao
UCD Department: Plant Sciences
UCD Mentor: Dr. Emilio Laca

Medusahead (Mh, Taeniatherum caput-medusae, (Sim). Nevski, Elymus caput-medusae), a noxious, invasive, annual grass, has invaded millions of acres in west coast states and is destructive to the natural ecosystem. Mh invasions decrease biodiversity, commercial and wildlife grazing value, and the capacity and recreation value of rangeland. Mh has a maximum period of susceptibility in the spring to control methods, but the start and end point are unknown. Thus, control methods, such as mowing, grazing, fire and herbicides, are ineffective or not feasible. The purpose of this project is to find the end point in that period to improve the timing of controls. First, a classification of partial Mh phenology, from post-pollination to complete maturity, was constructed. Then, seeds of different life stages from three different regions in California were germinated to find the germination percentages of each stage. The data, along with the results of a 1957 seed germination experiment, was then analyzed to identify the end point and to examine regional differences in Mh viability. The latest phenological stage at which Mh is still susceptible to control was found to be Intermediate 2. Also, differences in germination among the regions were identified. 

Tyrosine Sulfation in Voltage-Gated Potassium Channels

Student Name: Christina Ji
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Grace Rosenquist

Tyrosine sulfation is a posttranslational modification of a protein in which the hydroxyl group of the amino acid tyrosine is changed into a sulfate group. This modification strengthens protein-protein interaction. Tyrosine sulfation is prominent in the signature sequence of the conserved selectivity filter of voltage-gated potassium channels. The functionally similar KcsA from bacteria Streptomyces lividans shares this signature sequence. Positively charged toxin binding sites interact with negatively charged sulfated tyrosine sites. Voltage-gated potassium channels function in repolarization of action potentials in the brain, heart, and muscles. Tyrosine sulfation plays a critical role in the conduction of these channels.

The Identification of Bacteria Species in Vermicompost

Student Name: Gloria Liou
UCD Department: USDA Agricultural Research Services
UCD Mentor: Dr. Daniel Kluepfel

Standard microbial culture-based methods were used to study the composition and diversity of three vermicompost samples (Terra Vesco, Sonoma, CA, USA). Bacterial colonies were isolated from vermicompost using serial dilutions. The amplification and sequence analysis of 16S rDNA from 39 isolates showed a high percentage (79%) of Actinobacteria sp., especially Arthrobacter sp. (54%) and Streptomyces sp. (15%). Proteobacteria and Firmicutes sp. were also detected. The level of bacteria diversity in the soil supported the idea that microorganisms in vermicompost may be able to outcompete and reduce soil-borne pathogen populations, making vermicompost a viable disease-control method for the agriculture industry.

The Role of Genotype and Water Availability in Storage Carbohydrate Distributions in Grapevines

Student Name: Jennifer Chen
UCD Department: Plant Sciences
UCD Mentor: Dr. Kenneth Shackel and Dr. Mark Matthews

Perennial crop plants like grapevine (Vitis vinifera L.) use stored carbohydrates (CHO) from the previous season to sustain early season growth and flower development.  Water deficits inhibit current season growth and yield of the current season and following season, but the effects on stored CHO are not known.  CHO assays were performed on two grapevine genotypes, Grenache and Syrah, across three different irrigation treatments. Specifically, CHO concentrations in basal nodes and internodes were assayed just after the onset of veraison, the onset of ripening, which has been shown to be the annual low point in storage CHO concentrations in grapevine. A series of hot ethanol baths were used to extract soluble sugars. After extraction, CHO samples were digested into glucose using an alpha-amylase and amyloglucosidase solution. A PGO enzyme o-dianisidine dihydrochloride solution was added to each sample for a color reaction. The plate was read using a spectrometer and optical density numbers were recorded and analyzed with a standard starch calibration curve. In general, Syrah had higher storage CHO concentration than Grenache, which suggested that it was more efficient with water. Across both varieties and all three irrigation treatments, internodes contained higher concentration of storage CHO. CHO distribution in internodes supported the hypothesized relationship: decreased water availability results in decreased concentrations of storage CHOs. These data suggest that water deficit and storage CHO concentrations are positively correlated in perennial crop plants like grapevine (Vitis vinifera L.).  CHO in the nodes did not show the predicted distribution. Syrah vines had lower midday leaf water potential than Grenache vines which can be attributed to their anisohydric tendencies. This information on CHO distribution and genotypic variation in response to water deficits is important for identifying water-efficient grapevines and for water-conservation in agriculture.

A Home Use Descriptive Analysis Study to Evaluate Body Washes

Student Name: Jennifer Mou
UCD Department: Food Science and Technology
UCD Mentor: Dr. Michael O’Mahoney and Dr. Rie Ishii

Panelists were given blind samples of body washes to evaluate at home on a seven point scale. They received sufficient training to be able to distinguish between product differences for each attribute of the body washes. The goal of this study was to improve on the current methods of analytical product assessment with a home use descriptive analysis study to reflect more realistic home use situations of body wash products.

Evolution of 2-Nitrotoluene 2,3-dioxygenase and Gene Manipulation of Catechol 2,3-dioxygenase in Acidovorax sp. Strain JS42

Student Name: Jessica Li
UCD Department: Microbiology and Molecular Genetics
UCD Mentor: Dr. Rebecca Parales

Acidovorax (formerly Pseudomonas) sp. JS42 is able to use the compounds 2-nitrotoluene (2NT) and nitrobenzene as carbon, nitrogen, and energy sources. In the first step of the degradation pathway, 2-nitrotoluene 2,3-dioxygenase (2NTDO) converts 2NT to 3-methylcatechol with concomitant nitrite release. 2NTDO has three component proteins, and although it can break down all three mononitrotoluene isomers, JS42 cannot grow on 3-nitrotoluene (3NT) or 4-nitrotoluene (4NT). However, it is possible to force JS42 to adapt to 3NT and 4NT with resulting changes to 2NTDO. In this study, we evolved 4NT+ JS42 strains to grow on 3NT using long-term selection. Sequencing of the ntdAc gene encoding the α subunit of 2NTDO oxygenase revealed substitutions at the amino acid position 204 on the active site. Mutations at positions 238 and 248 were conserved from the parent 4NT+ strain and no other mutations were present, supporting the hypothesis that the amino acid position 204 is crucial for the evolution of JS42 to utilize new substrates. The next step of the 2NT degradation pathway involves catechol 2,3-dioxygenase (CDO). Both ctdE1 and ctdE2 encode CDO, and each has an associated regulatory gene (ctdR1 and ctdR2). Based on the phenotype of a mutant strain with deactivated ctdE1, ctdE1 seems critical to the degradation pathway. We will complement the ctdE1 mutants to verify the role of the enzyme in 2NT degradation. It is not yet known whether ctdR1 encodes an inducer or repressor, so ctdR1 inactivation will be used to determine the function of the regulatory protein. 

Effects of Cage Density on the Microenvironment and Health of Mice

Student Name: Jessica Ye
UCD Department: Anatomy, Physiology, and Cell Biology (Vet Med)
UCD Mentor: Dr. Kent Lloyd and Dr. Kristin Evans

According to the Institutional Animal Care and Use Committee (IACUC), Public Health Services (PHS), and Office of Laboratory Animal Welfare (OLAW), institutions must follow guidelines set by the Guide for the Care and Use of Laboratory Animals in their animal care and use programs when establishing mice populations within a cage to maintain the health and welfare of the laboratory mice2. The effects of mouse-caging density were studied in this project to determine the maximum number of mice that could be housed in a cage and maximum time between cage changes while maintaining the health and welfare of the mice. The CO2 level, Ammonia level, change in air flow, temperature, humidity, and any social, behavioral, or physical changes of the mice were evaluated. Seventy cluster of differentiation 1 (CD-1) outbred female adult mice were divided into four groups, with a minimum of two mice and a maximum of five mice per cage. Each week measurements of the microenvironmental gases in the cage, observations of the behavior and physical changes in the mice, cage temperature, cage humidity, cage airflow, and weights of the mice were recorded, for three weeks.

The Effect of Restoration on Soil Organic Carbon in Californian Grasslands

Student Name: Kaitlyn Gee
UCD Department: Plant Science
UCD Mentor: Dr. Emillio Laca

The atmospheric carbon sequestration abilities of plants, and in specific of grasses, are important on a local and global scale. However, little is known about the difference in carbon sequestration rates of perennial and annual grasses. The goal of this study is to discover if and to what extent grassland restoration, which is commonly achieved by seeding native perennial grasses, increases carbon sequestration through the analysis of soil organic carbon (SOC) stocks. Two sets of soil samples were taken, one set with samples from restored and nearby unrestored sites, and one with pairs of adjacent annual and perennial samples. The soils were processed and analyzed for carbon content in external laboratories. Statistical analysis using JMP revealed that the unrestored sites generally have higher amounts of SOC, although there is a positive relationship between perennial grasses and carbon presence. We conclude that the varying management practices and land use histories greatly shape SOC stocks in grasslands and that there is preliminary evidence that perennial grasses are more effective than annuals in storing carbon. 

Purification of Functional Milk Oligosaccharides for Human Health Improvement

Student Name: Kevin Chen
UCD Department: Food Science and Technology
UCD Mentor: Dr. Daniela Barile

Milk oligosaccharides play a prebiotic role in the growth and establishment of a balanced gut flora in infants, selectively enriching the beneficial and protective bacteria that promote healthy immune and cognitive development. By acting as anti-pathogenic agents, milk oligosaccharides hold great promise as high quality supplements that can help support the health of any population with comprised immune systems. In order to provide sufficient quantities of oligosaccharides for clinical and functional testing, this work optimized several steps of activated carbon oligosaccharide filtration from human milk permeates. For measurement of process efficiency, a carbohydrate assay was first developed by treating milk products treated with Carrez clarification. Carbohydrate adsorption tests on activated carbon followed by total carbohydrate and lactose quantification showed that ratios of activated carbon to carbohydrate from 5-10 had better carbohydrate retention. In addition, permeates filtered with finer mesh charcoal (20-500) at a charcoal/carbohydrate ratio of 10 had the majority of carbohydrates adsorbed. Optimized conditions found in this study can be applied to purification from bovine milk whey permeate, a dairy industry waste stream. With further refinements to other steps of the purification technique, activated carbon filtration can be adapted as an effective method for mass extraction of health promoting oligosaccharides.

Optimization of gram-scale synthesis of UDP-galactose via one-pot multienzyme approach

Student Name: Kevin Feng
UCD Department: Chemistry
UCD Mentor: Dr. Xi Chen

Uridine 5’diphopho-α-D-galactopyranoside (UDP-Gal) is a sugar nucleotide commonly used by animals, plants, and bacteria as the activated donor of galactose (Gal).   The salvage pathway is the simplest route to synthesize UDP-Gal and involves three different enzymes: Streptococcus pneumoniae TIGR4 galactokinase (SpGalK), Bifidobacterium longum UDP-sugar pyrophosphorylase (BLUSP), and Pasteurella multocida inorganic pyrophosphatase (PmPpA)1.  The optimum conditions for the entire reaction pathway in a one-pot multienzyme system were determined.  Then, using these conditions, synthesis of UDP-Gal was scaled-up to the gram-scale.  Production of UDP-Gal was successfully attempted using 1 g of Gal and a reaction yield of 70% was obtained.  No isolation yield was determined because of incomplete purification. 

The Investigation of the Genetic Basis of the Shade Avoidance Response in Arabidopsis thaliana

Student Name: Kisha Thayapran
UCD Department: Plant Biology
UCD Mentor: Dr. Julin Maloof

Shade avoidance is a series of plant responses elicited when a plant experiences a reduced ratio of red to far-red (R:FR) light. In this study we investigate the genetic basis of shade avoidance by examining hypocotyl and root growth of Arabidopsis thaliana in four natural variants and three mutant lines. The four natural variants were Columbia (Col), Kondara (KON), KNO-18, and RRS-10. The three mutant lines, numbered 19, 38, and 52, were TDNA lines with insertions in the gene of interest. The average hypocotyl length was found to be longer in the shade conditions for every genotype. From the difference between the average hypocotyl length in sun and shade conditions, it was found that 19 may be a high responder, 38 responds similarly to wild-type Col, and 52 may be a low responder. After 3 days of shade treatment, there was little difference between the average root length between genotypes or between plants grown in sun or shade conditions. However, primary root length measured after 10 days of light treatment revealed that the average root length was longer in the shade conditions for every genotype. This suggests that roots show increased growth in response to shade. These results are relevant because knowledge gained from this experiment can be utilized in the agricultural industry when deciding crop spacing and land conservation.

Examination of posttranscriptional regulation and de novo transcription during early embryonic development using RNA-Seq

Student Name: Krishna Bharathala
UCD Department: Animal Science
UCD Mentor: Dr. Pablo Ross

Background: Embryonic genome activation (EGA) is a critical time during mammalian development during which an embryo initiates transcription of many important genes after a long period of transcriptional silence and dependence on stored transcripts in the oocyte. In bovine, EGA typically occurs at the 8-16 cell stage. Before EGA, the oocyte and early embryo regulate protein production using post-transcriptional mechanisms like cytoplasmic poly-adenylation.  The global targets of this regulation are not very well understood. Despite the fact that the oocyte is mostly transcriptionally silent, there is some evidence of de novo transcription between fertilization and EGA.  In fact, it has been suggested that there is a minor EGA earlier in development at the 4-cell stage. This study focuses on that transcription and on the post-transcriptional regulation of stored maternal mRNAs. Results: Our results support and characterize the notion of EGA. They yielded inconclusive results with respect to cytoplasmic poly-adenylation and post-transcriptional modification. They also suggest that androgen receptors that play a role in oocyte maturation may also play a role in signaling between embryos and the mother during early embryonic development.

Identification and Quantification of Fusarium oxysporum f. sp. vasinfectum Infestation in Soil with a Real Time Polymerase Chain Reaction Assay

Student Name: Lauren Banks
UCD Department: Plant Pathology
UCD Mentor: Dr. Mike Davis

A real-time PCR protocol was developed to detect specific concentrations of Fusarium oxysporum f. sp. vasinfectum (FOV) race 4 in soil samples, using publicly available primers specific to FOV race 4. Environmental samples of equivalent spore concentrations from a range of 10^3-10^6 spores/ml were detected with this protocol. Soispore concentrations in this range have been shown to cause Fusarium wilt; this protocol detected both artificially spiked samples as well as environmental samples within this critical range. Pending further validation, this protocol will serve as an appropriate means of testing soil and determining soil spore load.

Determining the Pathway in which a Precursor Imports Through the Chloroplast Membranes

Student Name: Maya Lopez-Ichikawa
UCD Department: Plant Biology
UCD Mentor: Dr. Stephen Theg

Proteins transport across the chloroplast membrane through the translocon of the outer and inner membranes (Toc and Tic, respectively). In previous studies, the translocon was successfully plugged with a biotinylated precursor, RSSU-HC, bound to a large avidin molecule. Recently, the precursor was systematically shortened to find the minimum number of amino acids for a protein to span both membranes. From this, researchers can determine the protein import pathway and whether they import in a linear or folded manner.

Exploring MAP Kinases’ Role in Stress Response

Student Name: Neil Shieh
UCD Department: Plant Biology
UCD Mentor: Dr. Katie Dehesh

Plants are subject to many stresses daily.  In order to efficiently deal with all of these stresses, plants have devised a general stress response which allows them to immediately respond before executing a specialized response to deal with the stressor.  Multiple biological pathways in cells have been observed to activate the Rapid Stress Response Element (RSRE), which is the cis-element that plays a part in activating the general stress response.  Previous research conducted on the effects of various chemicals on RSRE activation indicated a possible relationship between Mitogen Activated Protein Kinase (MAPK) pathways’ and RSRE activation.  The individual effects of MAPK inhibitors on wound response, pathogen response, and basal response in one week old Arabidopsis thaliana seedlings will give a clearer picture of how MAPK pathways are utilized in those responses. This study shows that MAPK modulating chemicals are able to enhance specific stress responses. Additionally, one MAPK modulator was able to delay all RSRE induction.  Since MAPK pathways are conserved in animals, studying MAPK pathways’ role in plants could be applicable to human stress responses.

The Relation Between Memory Functioning and Anxiety and Depressive Symptoms in Children

Student Name: Nicolle Iacobacci
UCD Department: Psychology
UCD Mentor: Dr. Simona Ghetti

Previous studies have demonstrated a correlation between anxiety and depressive symptoms and memory functioning ability in adults (Castaneda, Tuulio-Henriksson, Marttunen, Suvisaari, & Lönnqvist, 2008). However, these studies have not yet been conducted on children ages 7 – 11, thus it is unclear as to whether or not there are any correlations between these symptoms to memory ability. This study will observe and examine how both anxiety and depressive symptoms correlate between memory ability and functioning in children. These data supported a relationship between both increased atypical internalizing behaviors and decreased memory abilities and they explore a possible relationship between externalizing behaviors and memory ability, but ultimately did not support any existing link between them. This new research gives parents a broader insight for their young children, so they can be aware of the many ways in which anxiety, depression, and other internalizing problems may affect seemingly unrelated areas of their child’s growth and development. 

Tobacco mosaic virus modifies the host gene expression and methylation in Arabidopsis

Student Name: Philip Hwang
UCD Department: Plant Pathology
UCD Mentor: Dr. Bryce Falk

The mutants being used in this research are named based on the gene that does not function in the plant. These mutants include the MET-1 mutant, RDR2 mutant, DCL3 mutant, and DDM1 mutant. These mutants are bing used because they are suspected of playing a part in A. thaliana’s virus-induced methylation pathway. The control being used in this research is known as the Columbia ecotype or Col-0. Arabidopsis thaliana is being used as the experimental plant because it is essentially the model plant for genetic research. The Tobacco Mosaic Virus (TMV) and Cucumber Mosaic Virus (CMV) were inoculated into A. thaliana in order to determine which virus can work with the Arabidopsis. First, a virus screening was used to determine which virus would work with the Arabidopsis thaliana with the use of a TMV and a CMV primer. Then after determining the virus strain that is most useful for the continuation of this research, total RNA is extracted and used to create cDNA. These cDNA sequences were then amplified using PCR with primers designed for certain genes in an analysis for gene expression in each of the Arabidopsis mutants. DNA is then extracted from the mutants and methylation sensitive PCR is then used to locate areas and patterns of virus-induced methylation. 

Effects of Non-Steroidal Anti-Inflammatory Drugs on Cell Oxidation and Proteasome Phosphorylation in Striated Muscle Cells

Student Name: Rajeev Parvathala
UCD Department: Western Human Nutrition Research Center
UCD Mentor: Dr. Betty Burri

Cassava is a root vegetable that is a primary source of energy for millions of people.  Unfortunately, the cassava root does not contain many nutrients, but it does contain varying levels of cyanide. Many cassava consumers are negatively affected by prolonged consumption of cyanide which can result in paralysis, ataxia, and other undesirable consequences to health. Current methods of cyanide reduction are too expensive or inefficient for the impoverished people that eat large amounts of cassava. This study focuses on trying to find inexpensive and feasible methods of cyanide reduction through pH treatments. The cassava was treated, rinsed and then tested for cyanide by diffusion into water. The water was then tested for cyanide using the La Motte Cyanide in Water Test Kit. It was discovered that weak acids and bases cause the greatest loss in cyanide. This research can be used for innovation of new cassava processing methods. 

Effect of Leaf Water Potential and Irrigation on Stomatal Conductance in Grape Vines

Student Name: Sam Leitess
UCD Department: Plant Sciences
UCD Mentor: Dr. Kenneth Shackel and Dr. Mark Matthews

Deficit irrigation, the practice of purposely providing plants with less than ample water, is used in some cultural practices. Deficit irrigation is believed by some to cause favorable conditions in vine yield and water use efficiency (Shackel 2011). This study aimed to plot the reaction of grape vines to varying irrigation treatments, specifically stomatal conductance (Gs) against transpiration rate (T) and midday Stem Water Potential (SWP). Gs was taken from three leaves on each vine at noon with a L-Cor 1600 porometer, T was taken every thirty seconds and averaged for each ten minutes with load cells, and SWP was taken once per day from each plant with a pressure bomb. Results indicated the time delay between last irrigation and vine reaction varies from vine to vine, but a general pattern of a sudden and large decrease in Gs several days after last irrigation was noted.

The Investigation of the Pathway and Production of Sugar Alcohols in Lactobacillus florum

Student Name: Sean Thomas
UCD Department: Food Science and Technology
UCD Mentor: Dr. Maria Marco

Erythritol is a naturally occurring, zero calorie sweetener produced by fermentation and is used as an additive in many foods and beverages. Erythritol is produced commercially through yeast fermentation using an expensive filtration process. The discovery of a microbe that could produce erythritol in comparable amounts could provide an alternative to this process, especially if it is a food grade microbe that could remain in the finished product. Lactobacillus florum is a novel, heterofermentative, lactic acid bacteria that has been found to produce erythritol making it a potential candidate.  Two strains of Lactobacillus florum, 8D and 2F, previously isolated from an unripe king palm fruit and a Valencia orange leaf, respectively, were tested for their ability and efficiency in producing erythritol when given glucose, fructose, or a combination of the two. Other metabolites, such as mannitol, acetic acid, lactic acid, and ethanol were also measured for. Since the erythritol pathway is not known in L. florum, enzyme assays were conducted to measure for the activity of enzymes responsible for erythritol production in Oenococcus oeni and yeast, erythritol 4-phosphate dehydrogenase and erythrose reductase, respectively. The results described will increase our knowledge of Lactobacillus florum, the pathway by which it can produce erythritol, and lead to areas of study for an alternative means of erythritol production. Both 8D and 2F were found to be capable of erythritol production when provided glucose, fructose, and the glucose-fructose combination, with 8D producing more than 2F.

Prediction of Sulfated Tyrosine Sites in Human Rhodopsin

Student Name: Shaan Somani
UCD Department: Neurobiology, Physiology, and Behavior
UCD Mentor: Dr. Grace Rosenquist

Tyrosine sulfation is a post-translation modification that has been proven to strengthen protein-protein interactions. In this study, we examine the potential for tyrosine sulfation in the human G-protein coupled receptor rhodopsin, the prototypic GPCR which plays a role in the visual phototransduction pathway in the rod cells of the eye. Through the position-specific scoring matrix, tyrosine sites Y26, Y29, Y30, and Y191 were found to have a high likelihood of sulfation in human rhodopsin. These sites were all well aligned in other forms of rhodopsin and cone opsins found throughout the animal kingdom. Y191, which is involved in a hydrogen bond network on extracellular loop 2 between E181 and Y192, was demonstrated through site-directed mutagenesis studies to function in the proper maintenance of Metarhodopsin II, the active state of photo-activated rhodopsin. Therefore, we find that sites Y26, Y29, Y30, and Y191 are consistent with sulfation.

Chlorpyrifos triggers mitochondrial dysfunction in mouse striatal neurons

Student Name: Viran Batth
UCD Department: Molecular Biosciences (Vet Med)
UCD Mentor: Dr. Cecilia Giulivi

Chlorpyrifos (CPF) is an organophosphate found in common pesticides that inhibits the enzyme acetylcholinesterase. Chronic exposure of CPF in humans results in neurological defects, developmental and autoimmune disorders, probably due to the generation of reactive oxygen species (ROS). Furthermore, CPF has been shown to have an effect on the dynamics and movement of mitochondria in rat cortical neurons and to decrease the activity of Complex I in PC12 cells. In this study, precursor neural cells from mouse brain were treated with 10 and 80 µM CPF for 24 hours. ATPase activity was found to be 20% lower on average compared to non-CPF-treated cells, with no changes in citrate synthase activity. Mitochondrial morphology and distribution were also analyzed through confocal microscopy. CPF-treated cells showed a significant decrease in the amount of tubular mitochondria (9% of untreated cells) accompanied by an 80% increase in the number of circular and fragmented mitochondria. Our results demonstrate that CPF exposure affects mitochondrial function suggesting its potential to produce oxidative stress and its involvement in oxidative stress-related neurodegenerative disease.

Prebiotics Help B. infantis Proliferate and Help Regulate Body Weight

Student Name: Won Park
UCD Department: Anatomy, Physiology, and Cell Biology (Vet Med)
UCD Mentor: Dr. Helen Raybould

The gastrointestinal tract (GI tract) of humans is colonized by immense amounts of microorganisms. They significantly affect the epithelial cells’ role of balancing the absorption of the necessary nutrients, ions, and water with the protection against potentially harmful toxins and pathogens. However, specific populations of microbiota tolerate only a certain range of conditions. Change in diet and nutrients (e.g. the consumption of prebiotics) can drastically change the population of microbiota present and consequently, the effects these microorganisms have on the GI tract.  This study suggests that inulin is able to help nurture B. infantis up to a certain point , that microbiota may play a role in regulating body weight, and that BMO may also be a prebiotic that helps microbiota proliferate.

2012
 

Investigating the Dof Gene in Tomato Plants

Student Name: Aditya Gupta
UCD Department: Plant Sciences
UCD Mentor: Dr. Diane Beckles

Aditya Gupta is researching the Dof gene in tomato plants. His research takes place in Asmundson Hall, which is one of the many buildings with the UC Davis Plant Sciences Department. His research will involve a series of tasks, such as amplifying the Dof gene and identifying the location where the gene is expressed on the tomato plant. Aditya’s research will eventually allow tomatoes to grow with 25% more yield and help tomatoes have higher nitrogen use efficiency. 

Understanding the Breakdown of Man Made Pollutants in Motile Bacteria

Student Name: Christie Ho

UCD Department: Department of Microbiology

UCD Mentor: Department of Microbiology

The lab seeks to understand the breakdown of manmade pollutants in motile bacteria, which are able to move towards the source of the pollutant and degrade the compound. Specifically, Christie is researching how a certain strain of bacteria can detect the presence of a specific compound, known as phenylacetate – a common compound found in many plastics and perfumes. Applications of this research include reducing the accumulation of environmental pollutants and minimize the effects of pollutants, helping preserve the planet. 

Detecting Viruses in Grape Leaves

Student Name: Divya Bhaskar
UCD Department: Plant Science
UCD Mentor: Vicki Klassen

Viruses in grapevine plants tend to be unevenly distributed which means a virus may be located in the basal (lower leaves) of the grapevine, or in younger leaves. This is a major problem for farmers all throughout California, because they do not know how many samples must be sent to the lab in order to be sure the results of the test are accurate.  The purpose of Divya’s project is to determine the incidence of false negatives in grapevines infected with one or more of the five viruses. Divya will collect infected plant samples on the UC Davis vineyards, and then she will process these samples in the lab using a variety of techniques and machines such as real-time PCR in order to extract the plants’ RNA. Divya’s project will occur in the Foundation Plant Science Department regarding viticulture under the guidance of researcher Vicki Klaasen.

Plant Mechanisms for Recognizing Invading Pathogens

Student Name: Kimberley Berg
UCD Department: Plant Biology
UCD Mentor: Dr. Dinesh-Kumar

For this project, Kimberley is working with Dr. Dinesh-Kumar in the Plant Biology department at the University of California, Davis. The lab studies the mechanisms by which plants recognize invading pathogens, initiate a signaling pathway, and execute a defense response. In general, the research aims to gain a better understanding of plant immunity. This information could be used to prevent plant disease in the agricultural market, promoting higher productivity of food products for the ever-growing human population.

Investigating Plant-Pest Interactions in Alfalfa

Student Name: Nikhil Jois
UCD Department: Department of Plant Sciences
UCD Mentor: Dr. Dan Putnam

Nikhil’s research involves alfalfa plants and plant-pest interactions, looking to control plant pests through the synthesis of novel phenolic compounds. It is currently hypothesized that these phenolic compounds found in plants deter pests by acting as prooxidants in the guts of the pests and causing cell damage. This work has a direct effect on forage production and pest control in an agricultural setting. The research may lead to further application in fruit and vegetable production as well

Determining the Effects of Environmental Tobacco Smoke on Rats

Student Name: Sarah Dukes-Schlossberg
UCD Department: Center for Health and the Environment
UCD Mentor: Dr. Kent Pinkerton

Sarah Dukes-Schlossberg is working at the Center for Health and the Environment at the University of California, Davis. She is working with Kent EPinkerton, Ph.D. of the School of Veterinary Medicine. She is researching the effects of environmental tobacco smoke (ETS) on the lungs of rats. Dukes-Schlossberg is comparing the lung tissues of spontaneously hypertensive rats (SHR) with those of control rats, SHR is a model for cardiovascular disease in humans. The results of the study will be used to better understand the effects of ETS on the lungs of humans and to develop better preventative measures and treatments.

Evaluating Symptoms of BRDC in Bovines

Student Name: Amy Kim
UCD Department: Department of Pathology, Microbiology, and Immunology
UCD Mentor: Dr. Laurel Gershwin

Kim is participating in a study on the bovine respiratory disease complex (BRDC), a deadly disease which costs the US cattle industry at least $3 billion dollars each year. Amy’s work in the lab, mainly involving virology, animal science, and molecular biology, will also include introducing different viruses to bovine steers for the evaluation of symptoms and for the search of traits that show resistance to this deadly bovine disease. In the end, not only will she will accumulate hours and hours of relevant lab experience, Amy will also help to find a cure to this costly disease

Determining the Structural Changes in Proteins During Staph Infections

Student Name: Andrew Kim
UCD Department: Chemistry
UCD Mentor: Dr. Donald Land

Andrew’s project is to determine the structural changes of the protein responsible for staph infections when incorporated in lipid vesicles that mimic cell membranes.  After creating the vesicles, he determines if the lipids maintain their spherical shape and the protein structure using infrared spectroscopy, a technique that analyzes absorbed light.  This project will allow researchers to gain a better understanding of how proteins interact with cell membranes.

Determining the Affect of Diabetes on Rat Cells

Student Name: Eric Gu
UCD Department: Molecular Biosciences
UCD Mentor: Dr. Cecilia Giulivi

Examining how diabetes has affected the shape of certain parts of the cells of the rats. One group of rats has had a gastric bypass performed to delay the onset of diabetes, whereas the control group will develop diabetes normally without the surgery. The researchers will then harvest the cells from the heart and muscles of the rats after specific time intervals. Hopefully, by studying how the mitochondria are affected, the team will be able to provide more insight as to how diabetes affects the body.

Creating Molecular Pesticides Against Olive Fruit Flies

Student Name: Irene Lin
UCD Department: Department of Chemistry
UCD Mentor: Dr. Dean Tantillo

Creating a pesticide against the olive fruit fly, which damages olives and affects the quality of olive oil. Irene is working on this project by using computer software to build molecules and test how well they work. An effective pesticide would have a large positive impact on the California olive oil industry, which produces a majority of American olive oil.
Two parental and 16 F2 offspring nematode lines were studied using polymerase chain reactions, restriction enzyme digestions, and agarose gel electrophoresis. Analysis of the final banding patterns allowed for comparison of parental and offspring mitochondrial DNA.
Results showed that paternal mitochondrial inheritance did not occur in any of the offspring lines. Even so, the limited sample size makes further experimentation necessary in order to conclude whether ever paternal inheritance occurs. Even more analysis is required to determine whether nematodes’ amoeboid sperm is responsible for this phenomenon.

Determining the Effects of Allergens and Air Pollution on Mice

Student Name: Kelsey Green
UCD Department: Center for Health and the Environment
UCD Mentor: Dr. Kent Pinkerton

Studying the effects of allergens and pollution on mice. Since asthma is actually the body attacking itself through an immune response of antibodies to allergens, Kelsy will be able to dose mice with allergens and pollution to see their effects on the lungs. She will look at slides of exposed mice lungs with a microscope, analyze mice for indications of inflammation, and analyze a specific protein using immunohistochemistry to determine the damaging effects of air pollution in the lungs and how this may exacerbate allergic asthma. The results will be to see how these two factors of allergens and particulate matter affect those with asthma and the mechanism of asthma. 

Investigating and Characterizing Protein Synthesis in Cardiovascular Disease

Student Name: Kunal Shah
UCD Department: Neurology, Physiology and Behavior
UCD Mentor: Aldrin Gomes

Protein synthesis is an important facet of any biological process. In the cardiac muscle, it is especially important in determining the state and condition of the muscle. One cardiovascular condition that is closely related to protein synthesis is cardiac hypertrohy. Cardiac hypertrophy is a thickening of the heart muscle, which results in a decrease in size of the chamber of the heart, including the left and right ventricles. This hypertrophy is a common known response to the increased workload of the heart that is associated with hypertension. The larger heart and cardiac cell mass means that more protein is necessary to make up the additional mass of the heart. This project investigated which proteins are specifically synthesized for the sake of contributing to cardiac hypertrophy and exactly how the rate of protein synthesis increases when cells are induced with hypertrophy by the cancer drug Doxorubicin. Using Click-iT AHA Protein Labeling Kits, and SDS-PAGE Gels, we were able to determine that the rate of protein synthesis increases by an average of 30.26%. In addition, we determined that Meclofenamate sodium, a Non-Steroidal Anti-Inflamatory Drug (NSAID), decreased the rate of protein synthesis in cardiac cells while simultaneously decreasing protein degradation, an important step in determining how exactly NSAIDs affect cardiotoxicity and induce cardiovascular disease. 

Using Brachypodium as a Model for Drought Tolerance in Monocots

Student Name: Lucy Cui
UCD Department: Plant Sciences
UCD Mentor: Dr. Eduardo Blumwald

Lucy Cui is working with Professor Eduardo Blumwald in the Plants Science Department to genetically engineer crops to be more drought and salt tolerant.  A model plant, Brachypodium, is used in place of wheat, rice, corn, barley, oats, and other monocots, to determine which genes to turn on in order to prolong crop life and produce greater yields with less water.  Another model plant,Arabidopsis, is used to understand the effect of genes that control pH in dicots, or flowering plants, on plant development. With the growing human population and loss of quality agricultural land, it is important to create these crops in order to continually feed the demanding population despite the harsh, arid conditions and to efficiently use the limited amount of agricultural land.  

Exploring the Effects of Photodegradation on Soil Organic Matter

Student Name: Nelson Chou
UCD Department: Plant Sciences
UCD Mentor: Dr. Johan Six

Nelson Chou conducts a research project in the Department of Plant Sciences. The mentors of this young scholar are Dr. Johan Six and Dr. Sandra Yanni of the Agroecology Lab group. The objective the research is to explore the effects of photodegradation on littler and soil organic matter and its magnitude and rate in a California grassland. Photodegradation is an abiotic process in which sunlight, composed of both ultraviolet and visible light, degrades organic materials and can therefore facilitate subsequent microbial decomposition of organic matter. The carbon and nitrogen amounts of soil, grass litter, light effect on decomposition will be strategically analyzed. These understandings will form a basis of accurate prediction of annual decomposition rates of highly sun-exposed ecosystems such as the California grasslands. 

Synthesizing the Protein MUC1

Student Name: Nicholas Andrew
UCD Department: Department of Chemistry
UCD Mentor: Dr. Xi Chen

Synthesizing MUC1, an important protein in human epithelial cells that protects cells from external disease. This protein is overexpressed in the cancer cells of the colon, ovaries, lung, and pancreas. By synthesizing various forms of this protein, Nicholas and his co-workers hope to begin developing a vaccine that can target and help the immune system destroy cancer cells while leaving normal cells intact.

Determining the Interaction of Plants and Indigenous Fungus

Student Name: Nicky Meyer
UCD Department: Plant Sciences
UCD Mentor: Dr. Sharon Strauss

Determining the interaction between plants and the fungus that live inside of plants. Through various experiments in the field, Nicky is looking to determine what impact the fungus has on the plant’s health, as well as its effect on the entire plant and animal community. Understanding the interaction between fungi and plants could lead to advances in agricultural techniques and environmental sustainability. 

Studying Tyrosine Sulfation as a Model for Cellular Response Activation

Student Name: Nikhil Kotha
UCD Department: Neurology, Physiology and Behavior
UCD Mentor: Dr. Grace Rosenquist

Some viruses have a special family of proteins (GPCRs) on their coat which sense specific molecules and bind to them thus activating a response inside the cell. Tyrosine sulfation is a modification to a tyrosine amino acid in the DNA sequence which strengthens this interaction between the protein and its target. As part of the Physiology Department at UC Davis and under the guidance of Dr. Grace Rosenquist, I am using methods such as 3-D modeling, programming, DNA sequence alignment, and an algorithm matrix in order to find the probability of a tyrosine amino acid, in the DNA sequence that codes for this protein, being sulfated and the importance and function of this protein. If a tyrosine site in a protein sequence has a high chance of being sulfated, this means that the protein and consequently the virus strength are being enhanced by tyrosine sulfation. I am investigating over 5 different viruses for identifying and pinpointing this phenomenon and future applications of my research involve potential drug research and development in order to stop tyrosine sulfation at the specific place where it happens and thus inhibit the virus from doing any further damage! 

Determining the Role of Viruses in Bovine Respiratory Complex

Student Name: Jason Manley
UCD Department: School of Veterinary Medicine
UCD Mentor: Dr. Laurel Gershwin

Manley’s research project deals with a bovine respiratory disease complex, a group of diseases that together kill millions of cows in the United States each year. Jason is studying a specific virus in this disease complex, called BRSV, to see whether a drug works as scientists hope in combatting the virus. If this drug is effective in cattle, further research could be pursued to determine whether the drug would work successfully against a similar virus that infects humans.

Characterizing the Efficiency of Catalysts

Student Name: Stacey Jeong
UCD Department: Department of Chemical Engineering and Materials Science
UCD Mentor: Dr. Bruce Gates

The research work is based on finding the characterizations of different catalysts to determine the most effective and appropriate catalyst. This will be done through various methods, such as IR spectrometry with unreactive gases to observe how much the spectra of the catalysts change. This work can be applicable to industrially used catalysts, which accelerate production and creation of other chemical-based materials.

Determining the Health Effects on Human Lungs Due to Air Pollution Exposure

Student Name: Tiankun Lu
UCD Department: Center for Health and the Environment
UCD Mentor: Dr. Laura Van Winkle

Tiankun’s project uses animal models to simulate health effects of exposure to air pollution on human lungs and involves the quantification and comparison of cellular damage caused by certain particulate matter (PM) between animals with different ages and between different compartments of lungs. Based on the previous work of his professor and through his own research, he might be able to determine the harmful portion of air pollution and potential biomarkers for susceptibility to PM that could be used either to identify or to treat afflicted individuals. The findings might also serve as reference and guidance to the government in regard of making environmental regulations and health policies.

Determining Biocontrol Methods for Bot Fungi

Student Name: Tommy Chiou
UCD Department: Plant Pathology
UCD Mentor: Dr. Doug Gubler

Tommy is currently working with Dr. Doug Gubler at the UC Davis Department of Plant Pathology, and his research involves several strains of fungi from the genus Botryosphaeria. His project tests several potential biocontrols, or methods of controlling pathogens through natural means, against the Bot fungi and seeing if any of them have inhibitory effects on the fungi. Fungi in the genus Botryosphaeria have been known to cause tree and shrub diebacks and diseases in grapes, apples, and pears. Each year, millions of dollars in revenue are lost in the agricultural industry because of damages by the fungi. Finding a cost-efficient and environmentally friendly method to control these organisms will greatly benefit the agricultural community. 

Determining the Role of microRNA During Early Embryonic Development

Student Name: Tony Hua
UCD Department: Animal Sciences
UCD Mentor: Dr. Pablo Ross and Juan Reyes

Researching the role of microRNA during early development of embryos. MicroRNAs are small RNA molecules that regulate gene expression post-transcriptionally and play a key role in diverse biological processes, including development, cell proliferation, differentiation, and apoptosis. Consequentially, altered microRNA expression is likely to contribute to human disease, including cancer and HIV. By studying the mechanisms that are caused by microRNA function, additional insight and a deeper understanding of related processes will allow for the pursuit of further research and possibly more efficient treatments or cures. Currently, the success rate of In Vitro Fertilization or fertilization outside the body is only at a 30% success rate for females age 35. By studying the regulatory roles of microRNA, processes such as In Vitro Fertilization could potentially benefit from increased success rates from a greater insight of correlated processes and techniques

Determining the Gene Control of Stress Hormones in Plants

Student Name: Yuki Koide
UCD Department: Plant Biology
UCD Mentor: Dr. Dehesh

Locating a gene that controls the expression of stress hormones in plants. This research will help current understanding of various stress responses in plants and what they have in common. It might be applied in the future to genetically engineer plants that are resistant to multiple stresses, such as drought or insect attacks.

2011
 

Movement of dissolved reactive phosphorous from soils to bodies of water in relation to eutrophication

Student Name: Kristen Chinn
UCD Mentor: Emily Carlson

Dissolved reactive phosphorous (DRP), or soluble reactive phosphorous (SRP), is a nutrient that can flow from agricultural land to bodies of water, causing eutrophication, which poses a threat to the health and efficiency of waterways. In order to prevent eutrophication from harming bodies of water, it is crucial to understand the movement of these nutrients, which was the main objective of this research.
Levels of DRP/SRP were analyzed by studying the sorption and desorption capacities of soils from different areas using the Bridgham et al (2001) method. This study found that sorption and desorption capacities varied between soil samples taken at different locations, indicating the level of effectiveness of different areas at retaining high amounts of phosphorous while releasing very low levels.
The information from this study can be used to evaluate potential areas for restoration based on the soil’s sorption and desorption abilities. In restoring wetland buffers, it is possible to benefit the environment by protecting water quality, using agricultural land more efficiently, managing waterways, and reducing or preventing eutrophication.