UH Faculty, Staff, and Student Works
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Browsing UH Faculty, Staff, and Student Works by Department "Biology and Biochemistry, Department of"
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Item A Black Death: Can Relic Oysters Be Used in Restoration Efforts?(2018-10-18) Sanchez-Ruffra, RachelThe eastern oyster, Crassostrea virginica, plays an important role in estuarine ecosystems, but populations have been decimated compared to historic levels. Large storm events, including hurricanes, periodically eliminate populations through freshwater input and sediment deposition, leading to death and entombment of shell in sediments. Buried within anoxic sediments, shells turn “black”, and with limited shell for restoration, resource managers have dredged this alternative substrate for larval settlement. Value for “black” shell to facilitate larval recruitment is unknown and this study examined the effectiveness of “black” sun cured shell, freshly removed “black” oyster shell, and normal restoration shell, taking into account background oyster population demographics. Oyster recruitment was significantly greater on sun cured black shell, algal growth on shells was ubiquitous among treatments, and larval recruitment was extremely limited. No significant difference was found from recorded oyster population characteristics, as they were similar to levels after initial construction. This suggests sun cured black shell can be utilized for restoration efforts, but immediate dredging efforts may be impractical. Further, the impact from large storms, such as Hurricane Harvey, can impact settlement substrate and larval supply, thus, resource managers should consider both factors when implementing future restoration efforts.Item A Historical Review of Hematopoietic Stem Cell Transplantation: Developmental Advancements Then and Now(2023-04-23) Wade, JoshuaA review of the history of regenerative medicine focusing on the use of hematopoietic stem cells (HSCs) from their initial inception in bone marrow transplantation for blood disorders and diseases to their expanded uses today through advancements in technology. Commonly treated malignant and non-malignant bone marrow transplant conditions include but are not limited to leukemia, non-Hodgkin lymphoma, and aplastic anemia. Bone marrow transplants are performed by autologous or allogeneic transplantation with multipotent HSCs. The first unrelated allogeneic transplant was performed by Dr. Edward Donnall Thomas in 1957. The initial approaches using stem cells as a form of treatment resulted in great difficulty and failure with few exceptions. However, discoveries such as the major histocompatibility complex within humans known as the human leukocyte antigen (HLA), allowed researchers to identify genetic matches between donors and recipients resulting in an increased rate of successful procedures. Since then, advancements in the field have introduced new techniques that improve the efficacy of hematopoietic stem cell transplantation and expanded use to more populations. Such advancements include improved conditioning regimens, matched unrelated donors, umbilical cord HSCs, and more potent immunosuppressants and antimicrobial drugs. For patients that experience tissue damage due to conditioning regimens or cancer malignancy, HSCs use a “homing” mechanism to mobilize to the affected area and release chemical factors that promote tissue recovery. Patients that undergo transplantation are put at risk of several conditions including graft-versus-host disease and opportunistic infections, however, complication-prevention regimens have been put in place to decrease the mortality rate. This literature review serves to roughly gauge how this field of medicine has developed since the 1950s and where future implications lie. Said implications include emerging ways to treat graft-versus-host disease, improved HLA typing matches, and HSC uses in transplantable organs like the liver.Item A long-acting Betaarrestin “biased” agonist to selectively activate AT1R to confer cardioprotection during heart failure and COVID-19 cardiovascular complications(2022-04-14) Medina, BryanIn patients with Heart Failure (HF) and COVID-19 induced HF, the principal hallmark is a depressed cardiac ejection fraction. Cardiac contractility is mediated through G-Protein Coupled Receptors (GPCR). A critical cardiac GPCR that mediates cardiac contractility is the Angiotensin Type 1 Receptor (AT1R). In the cardiovascular system, homeostasis is maintained by the Renin Angiotensin Aldosterone System (RAAS). An overactivation of the RAAS triggers an increase of Angiotensin II (AngII) and overactivation of AT1R, resulting in vasoconstriction, sodium retention and altered myocyte growth. These factors cause cardiac remodeling (ventricular hypertrophy and dilation) which overall leads to HF. Advancements in drug discovery are leading towards long-acting beta arrestin biased agonist that can simultaneously block AngII - mediated hypertension via AT1R-G alpha q-protein and activate cardioprotective AT1R-Betaarrestin. Activation of AT1R-beta arrestin will provide better therapeutic outcomes than other currently available non-selective drugs. At least two signaling pathways have been characterized for AT1Rs, G alpha q-protein and beta arrestin2. Signaling through the canonical pathway, AngII binds to AT1Rs to recruit Galphaq which causes cell injury. Alternatively, AngII binds to AT1Rs to recruit beta arrestin2 and induces cell protection. Our work will characterize the pharmacological properties of our synthesized peptide-based drugs for beta arrestin2 biased agonism to AT1Rs. We expect to develop a therapeutic drug that provides greater selectivity to ATR1-beta arrestinmediated signaling to increase cardiac contractility in failing hearts and prevent myocardial infarction in patients with COVID-19.Item A native Coreopsis variant competes more strongly against the invasive Verbena brasiliensis(2023-04-13) Nguyen, Minh KhueOrganic light- emitting diodes (OLEDs) are the top-performing technology for flat-panel color displays in smartphones and televisions. Cyclometalated iridium complexes are the most successful class of phosphorescent metal complexes used to produce light in OLEDs. Currently, blue-phosphorescent iridium complexes are the most challenging due to the requirement of the high HOMO-LUMO energy gaps. Strong s-donors are targeted as a supporting ligand for blue-emitting compounds. Acyclic diaminocarbene (ADC) and other strong s-donors ancillary ligands derived from isocyanide precursors are hypothesized to improve the phosphorescence efficiency. Understand the effects of these ancillary ligands on photophysical properties are the key to producing better blue-phosphorescent iridium complexes which are suitable for OLEDs application. Cycolometating ligands are used to control phosphorescence color and fluorinated 2-phenyl pyridine ligands are known to give blue phosphorescence. So fluorinated 2-phenyl pyridine-based cyclometalating ligands are used in this study and paired with different ancillary ligands installed by reactions between azide and isocyanides. Aryl isocyanide ligands with different electronic characteristics (CNAr-dmp, CNAr-NO2, CNAr-OMe, CNtBu) were used to produce different bis-cyclometaled iridium bis-isocyanide complexes. Then the bis-cyclometalated iridium bis-isocyanide complexes react with sodium azide (NaN3) in different solvents (CH2Cl2, CH3CN) at different conditions. This process produces new luminescent structures. These complexes have unique structures but were not obtained by the traditional synthesis route and they have unusual photophysical properties.Item A Proteomic Signature of Dormancy in an Actinobacterium: Micrococcus luteus(2017) Mali, Sujina; Rangel, Jonathan; Mitchell, Morgan; Havis, Spencer; Bodunrin, Abiodun; Khan, Arshad; Widger, William R.; Bark, Steven J.This project was completed with contributions from Arshad Khan from the Department of Pathology and Laboratory Medicine, UT Health Science Center, Houston.Item A Study of Cis-Regulatory Sequences Of Wnt1 in the Disease Osteogenesis Imperfecta(2021-05) Dalibalta, Maria SaadWnt1 is known to be mutated in the inherited cases of the disease Osteogenesis Imperfecta, a bone genetic disorder also known as the brittle bone disease. The canonical Wnt signaling pathway has a known role in bone homeostasis and development and studies have shown that wnt1, an evolutionarily conserved member of the Wnt signaling pathway, is particularly important for the Wnt canonical pathway in osteoblast differentiation. This suggests a potential role of wnt1 in bone homeostasis. Understanding the regulation of wnt1 expression by cis-regulatory elements during development is important to understand the role it plays in bone development and homeostasis. This project uses ATAC seq (Assay for Transposase-Accessible Chromatin) datasets available to the public to study regions within and surrounding the wnt1 gene to find open chromatin regions, as this is associated with enhancer activity. The enhancers that are shown to be active are studied closely using the ENCODE project to obtain information on the expression profiles of the enhancers and the tissue in which they may be active. The activity of the enhancers was compared in tissues where wnt1-signaling is known to be important: hindbrain tissue, adipose tissue, and bone marrow macrophage and two enhancers with the highest activity were identified. By knowing these cis-regulatory sequences of wnt1, now it could be easier to possibly use wnt1 expression as a therapeutic target in the disease Osteogenesis Imperfecta.Item ALCAM Deficiency Dampens Renal Inflammation in Lupus Nephritis(2022-04-14) Nune, RadhaSystemic lupus erythematosus (SLE) is a chronic, autoimmune disease affecting many organs. In nearly 50% of patients with SLE, the disease progresses to lupus nephritis (LN), a severe complication of SLE that increases the risk of end-stage kidney disease and mortality. There is no cure for LN, and the pathogenesis of the disease remains incompletely understood. In a comprehensive screening of over 1000 urine proteins, the activated leukocyte cell adhesion molecule (ALCAM) was identified as a top molecule elevated in the urine of patients with active LN compared to healthy controls. Expressed on antigen presenting cells, ALCAM serves as CD6 ligand. ALCAM binds CD6, resulting in recruitment of signaling factors such as SLP-76 and GADs that enhance TCR signaling. It also activates several MAPK pathways related to T cell activation, proliferation, differentiation, and infiltration into inflamed renal tissues. Such upregulated T cell activity contributes to end organ disease by promoting autoantibody production, subsequently leading to B cell differentiation and expansion. Despite its significant modulation of immunity, the precise role of ALCAM in the development of LN remains unknown. Given ALCAM's published function in autoimmunity, we examined whether the development of LN is contingent upon the presence of ALCAM. We anticipate that our study will shed light on ALCAM's role as a key pathogenic player and potential therapeutic agent in lupus nephritis.Item Alleviating the Maternal Mortality Epidemic Caused by Preeclampsia Through CHW Based Interventions(2019) Ahmed, SarahMaternal mortality rates in the US have been consistently rising and are currently higher than any other developed nation. Preeclampsia, a hypertensive disorder of pregnancy, affects about 18% of pregnancies worldwide [13], and 60% of preeclampsia deaths had a good to strong chance of being prevented [6]. A DAG (Directed Acyclic Graph) model was created to show causes and effects of preeclampsia and to see where in the pathway an intervention would be most effective. Stories were collected from women to get a perspective of the issue through the lens of those experiencing it. Based on the DAG model and the stories, a two pronged intervention was designed. One prong focused on health professionals while the other focused on the public. A simulated city modeled after Houston was created to see how many people were affected by the condition, how many deaths were expected, and how necessary the intervention was. On this basis, it was found that raising awareness and teaching the population how to be their own advocate, how to deal with medical professionals, and preventative measures they could be taking to prevent any complications was the best method to help decrease the number of preeclampsia-related deaths.Item Amino Acid purification and Single-Molecule FRET study of tRNA translocation(2019) Kapadia, Taher; Lin, RanUnderstanding the basic translational mechanisms of the ribosome during protein translation is essential to developing novel chemical defenses against the increasing amount of anti-biotic resistant bacteria. To study the fundamentals of the protein building process, we can employ smFRET studies which require debris-free, pure samples of amino acids, the building blocks of all proteins. Our experiment focused on the overexpression and purification of the Glutamic Acid tRNA and its translocation in the ribosome. This was accomplished by inserting a Glutamic Acid sequence into a vector plasmid through a series of digestion and ligation reactions. The recombinant plasmid was then overexpressed in E. Coli and the tRNA was purified through chloroform extraction, gel filtration, and ion-exchange chromatography. The pure sample was used to make a ribosome complex and the translocation of Glutamic Acid from the A site in the ribosome to the P site was studied through smFRET. From the results, it was concluded that the ribosome can efficiently process the Glutamic Acid tRNA when a frameshifting motif is not present. This raises the question of what may occur if frameshifting motifs are added into an mRNA transcript and how this may affect the trans-locational process of glutamic acid. The future of this study should target the translational process in the presence of frameshifting motifs to identify targets which can halt the building of the proteins. Studies attempting to apply such mechanisms to cells in a host should, however, be mindful that both the bacterial and host ribosomes are susceptible to frameshifting which may limit or complicate discovery of target sites.Item An Analysis Characterizing the composition of the Corneal Limbal Epithelial Stem Cell Niche (LSCN)(2022-04-14) Parsaie, ArianLimbal epithelial stem cells (LESCs) are required for the long-term maintenance of the corneal epithelium and are required to regenerate the corneal epithelium after injury.The loss or damage to LESCs or the LSCN can lead to Limbal stem cell deficiency (LSCD). LSCD causes corneal erosions, neovascularization, and conjunctivalization of the cornea, which leads to ocular pain and vision loss. Hyaluronan (HA) is a constituent of the LSC Niche (LSCN) that is necessary for maintaining LESCs in their stem cell state. My research this summer focused on characterizing the HA-specific LSCN by identifying HA-bound proteins and proteoglycans (PGs) within the limbal region of human and porcine corneas. HA bound proteins and proteoglycans (PGs) were extracted from the central corneas and limbal region. Firstly, we dissected the corneas and used chaotropic agents and mechanical trituration to homogenize the tissues. Then, an anion exchange chromatography was used to isolate HA bound proteins and PGs. Finally, samples were desalted and concentrated.Total protein/PG extracts were analyzed by Western Blotting and agarose gel electrophoresis. Additionally, human corneas were processed for histological analysis. Inter-Alpha Inhibitor (IaI), a molecule that forms specific HA matrices with anti-inflammatory properties, was identified in our samples by both Western blotting and immunohistochemistry. Future studies will work on characterizing the role of this HA/IaI or HA/TSG-6/IaI matrix within the corneal limbus. Characterizing the role of the LSCN is vital for establishing novel mechanisms for treating LSCDItem An Analysis of Colony Movement and The Effects of Movement on Fitness in the Western Harvester Ant, Pogonomyrmex Occidentalis(2023-08-07) Ramsaroop, MaxximusColony movement in ants (Hymenoptera: Formicidae), is well documented and several studies have explored the fitness costs and causes of colony movement in different species. Attempts to describe the purpose or patterns of colony movement have been made on many species within Pogonomyrmex spp. However, research into these aspects of colony movement has yet to be described in P. occidentalis, The Western Harvester ant. This thesis examines the fitness costs, potential causes, and behavioral syndrome associated with colony movement in P. occidentalis. I analyzed Dr. Cole & Dr. Wiernasz's data from 1993-2023 on colony movement, age, size, distances traveled, and survivorship for 6,066 colonies. Moved colonies were found to have a smaller colony size, shorter colony lifespan, and lower rates of survivorship when compared to unmoved colonies using t-tests and a Kaplan-Meier survivorship analysis. Moreover, moved colonies were compared against 1-year-old colonies, and were found to have a larger colony size. Colonies were also found to have a greater tendency to move later in age, and 16.6% of colonies had moved at least once. These results suggest that colonies that move incur fitness costs, as colony size and lifespan are proxies for fitness, and that colony movement is more costly for younger or smaller colonies. Furthermore, because P. occidentalis is notably long-lived, shows high nest fidelity, and their movements are associated with a loss in fitness, they likely exhibit an adventitious nest relocation syndrome. Through this thesis, we can better contextualize P. occidentalis in the greater scheme of animal architects and the decisions they make.Item An Analysis of Errors and Discrepenices in Analyzing Single Cell RNA Sequence Data(2021-05) Modi, Paulomi SanjaykumarSingle-cell RNA sequencing (scRNA-seq) is an extremely vital sequencing technology that has enabled High-throughput mapping of cellular differentiation hierarchies. scRNA-seq has excellent sequencing potential with a wide range of applications beyond regular transcriptome profiling. scRNA-seq process involves analyzing data using 3' end counting technology, which involves sample composition and analytical processing including pre-processing, normalization, alignment and clustering. In order to accomplish this task bioinformaticians around the world have developed many computational tools. As of 2019, there exist 385 different tools that can be used to analyze scRNA-seq data, and that number is growing. Although this continuous addition of new features to single-cell data analysis confronts technical gaps with bulk RNA-seq, there have been very few attempts to standardize these practices. This study explores the various approaches to re-analyze previously published single cell RNA-cell sequencing data and discusses subsequent challenges to utilize publicly available data sets to conduct a multicenter study. Considering the differences in data publication formats, there are several methods that can be employed. 1) Analyzing BCL files 2) Analyzing FASTQ files 3) Analyzing matrix files 4) Analyzing Seurat or ScanPy objects. This thesis provides a concise overview of some of the steps, algorithms, and approaches that are currently used in the analysis of single-cell RNA-sequencing data, with an emphasis on recent developments. Hence, I propose that in order to develop reproducible algorithms and analysis software for scRNA-seq data sets, it is vital that standardization across all analysis platform exist and the software developers recognize and understand the computational challenges posed by the analysis tasks.Item An Evaluation of Dobzhansky-Muller Incompatibilities in Protein Evolution(2022-05-05) Esopenko, HannahThis study is focused on the evolution of Dobzhansky-Muller Incompatibilities (DMIs) and Compensated Pathogenic Deviations (CPDs) in protein evolution. DMIs are genetic differences that occur by post zygotic isolation to reduce the overall fitness of an organism. Meanwhile, CPDs are pathogenic mutations that show no adverse effects to the organism as there is an additional mutation somewhere in the sequence that compensates for the deleterious nature of the mutation. Therefore, studying the nature of DMIs and CPDs provides a deeper understanding as to how deleterious events arise throughout the evolution of species. A study conducted by Kondrashov et al. (2002) addressed DMIs in protein evolution by identifying the occurrence of CPDs when the nonhuman orthologs deviated from the reference human ortholog sequence. Kondrashov et al.’s (2002) study was clever in construction, but the methodology was unclear, and the results appeared to be over simplified. To analyze the validity of the Kondrashov et al. (2002) paper, a similar study using restricted parameters and modern bioinformatic databases was conducted for this senior thesis project. To do so, 24 primate orthologs of 32 genes responsible for Mendelian diseases were collected and compared to the pathogenic missense data of humans to identify CPDs. Through computational analysis and the visual representation of protein alignments, 26 valid CPD hits were found. The 26 CPD hits presented in four general patterns: single species CPD, single clade CPD with two or more species, convergent evolution of a CPD, and ancestral CPDs. A statistical analysis was performed to determine whether factors such as the length of the protein, the evolutionary distance between sequences, or the number of pathogenic variants played a role in the number of CPDs found. The relationship between the number of CPDs found and the evolutionary distance between sequences and the amount of pathogenic variant data were found to be statistically significantly correlated. More data and research into primate genomes and the nature of CPDs is required to accurately determine their occurrence. This will help predict how CPDs arise in species and better evaluate the claims made in the Kondrashov et al. (2002) paper.Item An Investigation into Patterns of Scaling Among Morphological Traits(2018-10-18) Mai, SarahProper scaling of morphological traits to body size is necessary for ecological function. Individuals with better nutrition grow larger body sizes and traits, whereas individuals with poor nutrition develop smaller body sizes and traits. However, morphological scaling relationships may differ among traits. Fly wing size scales approximately 1:1 with body size, whereas traits such as beetle horns have a larger trait to body size ratio and insect genitalia has a smaller trait to body size ratio. Changes in the slopes of such scaling relationships result in morphological evolution, but the role of development in the expression of scaling relationships and how variations in development will influence the changes in scaling are unknown. Individual Drosophila melanogaster from isogenic lines were subjected to different diets in the larvae stage to produce the complete size range for each genotype. The wing and body size of these individuals were measured and used to fit genotype-specific scaling relationships. We will investigate the scaling of other morphological traits in the lineages that exhibit the steepest and most shallow slopes for wing: body size scaling, to determine if scaling across traits is correlated within a genotype. Such correlations may suggest that the same alleles regulate scaling for multiple traits, suggesting a restriction on the independent evolution of morphological scaling relationships.Item An Investigation into the Correlation between PM 2.5 and Low Birth Weight Rates in Texas(2020-09-29) James, Anjali; Jimenez, Ashley; Juhas, Claire; Mandviwala, Sakina; Sanati, SalarStudies from countries such as Brazil and the Czech Republic show pregnancy and infant abnormalities relating to an increased concentration of particulate matter (PM) 2.5 in the atmosphere for inhalation by mothers during the second trimester. Texas has been highlighted as one of the United States’ most polluted states due to excess petrochemical and manufacturing facilities, which emit PM 2.5 along with other carcinogens. This study evaluates the impacts of PM 2.5 emissions on the newborn population in Texas by comparing regional emissions data to reported Low Birthweight (LBW) Rates of newborns across Texas. Average PM 2.5 measurements are taken in micrograms per cubic meter (ug/m^3) throughout different regions where air monitors are distributed, sorted by county. Low Birth-weight Rates are identified as the average population of reported newborns under 2,500 grams in a given year per county. Furthermore, this study identifies the top PM 2.5 emission sites in Texas throughout the years of 2014-2018 and compares them to trends in their yearly county LBW rate. LBW has been linked to increasing complications with age including diabetes and heart disease. The findings of this study can provide further insight onto whether Texas, who receives one of the highest funding allocations for air quality monitoring and pollution control of any state by the EPA, should redistribute a bulk of its funding from monitoring to preventative measures in PM 2.5 pollution due to harmful links associated with these prenatal inhalants by mothers.Item An Unbiased Approach on The Conformational Dynamics of In-Solution Fibrinogen and Its Physiological Implications.(2022-04-29) Esparza Pinelo, Jose E.By itself, Fibrinogen (Fng) stands out as one of the most complex hematopoietic proteins in the cardiovascular system for multiple species in nature. Upon its activation and further cascade mechanisms, Fng can polymerize into fibrin and contribute to blood clot formation and substantial growth. Fng’s interactions with fibrinolytic proteins aggregate into a conglomerate of different fragments in blood bodily mechanisms. Any form of dysregulation in any of these pathways can lead to several complications not only within the cardiovascular system but throughout the entirety of the body. Understanding the crux of Fng’s functions and interactions with itself along with other proteins ultimately can be traced back to its inherent dynamic structure. In this study, I aim to probe the intrinsic flexibility that is beset on Fng by way of its multi-domain composition, allowing it to withstand incredible mechanical forces as well as being highly dynamic in its physiological form. Thus, extending the key biological concept that structure and flexibility that comes with it determine functions. Through an unbiased approach by implementing protein structural studies as well as computational dynamic simulations, in-solution Fng dynamics were studied in their totality.Item Analysis of Early Myelin Development in the Central Nervous System(2018-10-18) Sidiq, SaadMyelin, produced by oligodendrocytes (OLGs), is the major factor for the stability of neural transmission in the central nervous system (CNS). The interaction between axons and OLGs has been of interest in determining factors to increase myelination. This study aims to examine genes that may be responsible for this increase in early postnatal development. Expression level results show that genes responsible for mitochondrial transport protein stability and axonal outgrowth (MIRO1 and TRIO, respectively) were significant in early postnatal development, while the gene involved in OLG process extension through actin cytoskeletal rearrangement was significant in late postnatal development (ERMN). Results from this study can serve as preliminary data for others studies concerning factors affecting myelination, with applications to treatment of de-myelinating diseases such as multiple sclerosis. This project was completed with contributions from Philip Horner, Scientific Director for the Center for Neuroregneration at the Houston Methodist Research Institute.Item Androgen receptor mutation affects testes organization in an African cichlid A. burtoni(2022-04-14) Mathews, MarkAndrogen receptors (AR) are a ligand-dependent transcription factor that modulate the expression of androgen target genes. Androgens are hormones that bind to ARs and are essential to reproductive development and fertility. Past research illustrates that mutations in AR lead to small testes, disorganization of seminiferous tubules, and delay the process of spermatogenesis. The objective of this research is to investigate how the mutation of both ARalpha and ARbeta affect the organization of the testes and the process of spermatogenesis in the African cichlid fish, Astatotilapia burtoni. Firstly, we used a cryostat to section the testes at a thickness of 10 microns. Once the slides were dry, we conducted a Hematoxylin and Eosin (H&E) stain on the tissue in order to differentiate cell types. Photomicrographs were then taken of multiple sections in each testis and a grid of randomly generated points was overlaid. The cell type underneath 50 points was counted on a random section of the tissue; cell types of spermatogonia, spermatids, spermatozoa, and interstitial tissue were quantified as a measure of spermatogenesis. The homozygous ARalpha;ARbeta mutants appear to have numerous holes relative to the wild type (WT) testes and this will be quantified in the future. Furthermore, quantifications of cell types will be analyzed between WT and mutants in the future as well as the fertility of the sperm of the mutants.Item Antibody-Functionalized Nanophosphor Particles for Use as Reporters in Lateral Flow Assays(2017-10-12) Hasan, Aisha; Goux, Heather; Trabuco, JoãoThe main goal of this project was to identify important buffer components that improve the performance of nanophosphor reporters in a lateral flow assay. A model system based on the capture of the human chorionic gonadotropin (hCG) by two antibodies: monoclonal mouse anti-β hCG antibodies (Arista Biologicals, item # ABBCG-0402) and goat polyclonal anti-α hCG antibodies (Arista Biologicals, item # ABACG-0500). The principles of partial factorial design of experiments were used to screen common buffer additives. Comparing the relative effect of such components is key to develop better buffer formulations that enhance the performance of nanophosphor-based LFAs. Furthermore, relating the effects of certain buffer components to its mechanism of action could also shed a light on the interactions involved in the assay such as specific and nonspecific interactions or liquid diffusion through a porous membrane.Item Aspects Affecting the Expression of Hypoxia-Inducible Factors in Cardiomyocytes(2018-10-18) Vo, JenniferBy comparing the immunofluorescent images of each of the wells containing cells in different conditions, we can compare signals that are relevant to diabetic cardiomyopathy change HIF expression, activity, or localization. The cells were put in normal media, CoCl₂, or isoproterenol, and were transfected with HA HIF-1α and HA HIF-2α. We added only the 2nd antibody to one of the wells for a negative control. The signals from this well would show how much of the other signals should be expected as background. We knew from prior research that CoCl₂ should increase HIF expression and used cells in CoCl₂ as a positive control. The mouse HIF-1α antibody (ab113642) can be detected using cyanine 5 dye (CY5) because it emits a wavelength of 488. Similarly, the rabbit HIF-2α antibody (PA1-16510) can be detected using green fluorescent protein (GFP) due to its wavelength of 647. The results supported previous studies from other cell types. The cells in CoCl₂ were transfected with HIF-1α and HIF-2α overexpressed the HIFs in comparison to those in normal media. Additionally, we found that cells treated to induce hypertrophy also may increase HIF expression.