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Full-Text Articles in Biochemistry

Generation Of Neural Stem Cells (Nscs) From Human Fibroblasts Using Qq-Modified Sox2 And Neurod1 Proteins, Abdullah Ibrahim Alhomoudi Jan 2020

Generation Of Neural Stem Cells (Nscs) From Human Fibroblasts Using Qq-Modified Sox2 And Neurod1 Proteins, Abdullah Ibrahim Alhomoudi

Wayne State University Theses

The generation of induced neural stem cells (iNSCs) and induced neuronal cells (iNCs) from somatic cells provides new avenues for basic research and potential transplantation therapies for neurological diseases. However, clinical applications must consider the tumor formation capabilities of the implanted cells, the inability of iNCs to self-renew in culture, and reprogramming methods that use retroviral transduction which permanently alter genetic network of the cells. Here we report the generation of protein-induced neural stem cells (piNSCs) from human dermal fibroblasts using QQ-SON pluripotent reprogramming as a tool to quickly reset the time clock of the human somatic fibroblasts to a …


Characterization Of The Tsc/Dyrk1a Interaction, Supriya Joshi Jan 2020

Characterization Of The Tsc/Dyrk1a Interaction, Supriya Joshi

Theses and Dissertations

The Tuberous sclerosis complex (TSC) includes TSC1, TSC2 and the TBC1D7 subunits that together function as a principal inhibitor of the mTOR protein kinase complex 1 (mTORC1). mTORC1 is a master regulator of cell growth and proliferation that responds to signaling cues such as growth factors and nutrient availability. Proteomic studies in our lab revealed an interaction between the TSC subunits and DYRK1A, a ubiquitous protein kinase encoded by a gene located in the Down syndrome (DS) region on human chr21. In this study, we sought to validate the interaction of the TSC components with DYRK1A and to determine the …


Characterizing Glycosaminoglycan And Glycosaminoglycan-Oligo Interactions With Cancer Stem Cells And Their Regulators, Rahaman Navaz Gangji Jan 2020

Characterizing Glycosaminoglycan And Glycosaminoglycan-Oligo Interactions With Cancer Stem Cells And Their Regulators, Rahaman Navaz Gangji

Theses and Dissertations

Tumor metastasis and recurrence account for more than 90% of cancer-associated deaths and are thought to occur due to a small subpopulation of neoplastic cells known as cancer stem cells (CSCs). Because CSCs are resistant to conventional chemotherapy, there is a critical need to develop CSC-targeting agents. Microenvironmental components, especially glycosaminoglycans (GAGs), play a key role in regulating CSC phenotype. GAGs such as heparan sulfate (HS) interact with hundreds of proteins and influence a variety of biological processes including tumor proliferation, angiogenesis, metastasis and stem cell biology.

Recently, the Desai and Patel laboratories have demonstrated that a hexasaccharide form of …


Regulation Of Vascular And Lymphatic Endothelial Barrier Function By Sphingosine-1-Phosphate, Jacqueline Bellaci Jan 2020

Regulation Of Vascular And Lymphatic Endothelial Barrier Function By Sphingosine-1-Phosphate, Jacqueline Bellaci

Theses and Dissertations

Endothelial cells play an important role in maintaining homeostasis, and disruption of the endothelial barrier have been linked to a number of different pathologies, including cancer metastasis. The vascular endothelial barrier is known to be regulated by a variety of factors, including sphingosine-1-phosphate (S1P), a bioactive lipid metabolite, which has been shown to improve barrier function. However, most studies to date have been focused on the vascular endothelium, and the regulation of the lymphatic endothelial barrier has not been thoroughly investigated. We found that prior to any stimulation the lymphatic endothelial cells (LECs) had a lower trans-endothelial resistance (TER) in …


Structural And Functional Characterization Of Cazyme And Cazyme-Related Proteins From: Bacteroides Thetaiotaomicron And Porphyromonas Gingivalis: Two Abundant Colonizers Of The Human Microbiome, James Stevenson Jan 2020

Structural And Functional Characterization Of Cazyme And Cazyme-Related Proteins From: Bacteroides Thetaiotaomicron And Porphyromonas Gingivalis: Two Abundant Colonizers Of The Human Microbiome, James Stevenson

Theses and Dissertations (Comprehensive)

The human body consists of approximately 30 trillion cells, while non-human microbes that reside on and within the body outnumber human somatic cells by a factor of 1.3 – 2.3. The interplay between our cells and those of the colonizing microorganisms affect physiology in a multitude of ways, both beneficial and detrimental. Microbes found in the oral cavity, such as the Red Complex member Porphyromonas gingivalis, are associated with pathology, namely periodontal diseases including gum deterioration, tooth decay, and loss of underlying alveolar bone. At the other end of the gastrointestinal tract, microbes such as Bacteroides thetaiotaomicron are found …


Synthesis And Biophysical Analysis Of The Cell-Penetrating Peptide Penetratin And Its Aromatic Analogues, An Le Jan 2020

Synthesis And Biophysical Analysis Of The Cell-Penetrating Peptide Penetratin And Its Aromatic Analogues, An Le

Theses and Dissertations (Comprehensive)

Cell-penetrating peptides (CPPs) interact with biological membranes, undergo cellular intake/uptake, and may act as potential drug delivery agents. Understanding the molecular interactions of these peptides with membranes contributes to gaining a better knowledge of their potential use in medical and pharmaceutical applications to improve human health. The current research focuses on understanding the mechanisms of a CPP in interaction with different model phospholipid membranes. The peptide penetratin (primary sequence: RQIKIWFQNRRMKWKK) is an example of a CPP that can interact with and pass through biological membranes. The current thesis provides spectroscopic and calorimetric evidence that penetratin associates with lipid membranes. Circular …


Functional Characterization Of Proteins From A Putative Chondroitin Sulfate A Degrading Operon And The Crystal Structure Of An Exo-N-Acetylgalactosamine-4-Sulfatase From Tannerella Forsythia, Peter Nguyen Jan 2020

Functional Characterization Of Proteins From A Putative Chondroitin Sulfate A Degrading Operon And The Crystal Structure Of An Exo-N-Acetylgalactosamine-4-Sulfatase From Tannerella Forsythia, Peter Nguyen

Theses and Dissertations (Comprehensive)

Advanced periodontitis has been shown to have strong associations with the residence of a bacterial triad of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola also known as the red complex. These bacteria were previously thought to be asaccharolytic, making them better suited to survive in deep in periodontal pockets where access to carbohydrates from food is limited. Information gathered from genomic analysis suggested that upregulation of an operon (BFO2285-BFO2295, BFO3043) could confer the capacity for T. forsythia to degrade chondroitin-4-sulfate, the principle glycosaminoglycan of alveolar bone tissue. Bioinformatics analysis suggested the operon was similar to an operon found in Bacteroidetes …


Effects Of Seasons And Zonation On Biochemical Contents Of Two Seagrass Species Along Dar Es Salaam Coast, Tanzania, Wamwi M. Mbaga, Esther F. Mvungi Dec 2019

Effects Of Seasons And Zonation On Biochemical Contents Of Two Seagrass Species Along Dar Es Salaam Coast, Tanzania, Wamwi M. Mbaga, Esther F. Mvungi

Tanzania Journal of Science

Accumulation and allocation of biochemicals in plants are affected by various factors. The influence of seasons and vertical zonation on biochemical contents of seagrasses Cymodocea rotundata and Thalassia hemprichii from Mjimwema seagrass meadow along the coast of Dar es Salaam were assessed. Samples were collected in 2016 during wet/rainy (April–May) and dry (June–July) seasons. Concentrations of crude protein, soluble carbohydrates, phosphorus, lipid, and proline were determined using standard methods. Significant higher levels of phosphorus, lipid, protein and proline were observed during the dry season than in wet season (p < 0.05). Likewise, seagrass population occurring at the shore zone expressed higher levels of biochemical contents than those found at the subtidal zones. The highest concentration of protein was recorded in Thalassia hemprichii (5.8 mg/g dw)), while those of lipid and carbohydrates (27.1 and 53.1 mg/g dw, respectively) were recorded in Cymodocea rotundata. The two species studied did not vary significantly in their biochemical concentrations in both seasons. Generally it was observed that the concentrations of studied biochemicals were up-regulated during the dry season and varied depending on the locations where the seagrass grow. Thus, both season and zonation played roles in biochemical accumulation in the seagrasses understudy, however not interactively.


Robust Cullin-Ring Ligase Function Is Established By A Multiplicity Of Poly-Ubiquitylation Pathways, Spencer Hill, Kurt Reichermeier, Daniel C. Scott, Lorena Samentar, Jasmin Coulombe-Huntington, Luisa Izzi, Xiaojing Tang, Rebeca Ibarra, Thierry Bertomeu, Annie Moridian, Michael J. Sweredoski, Nora Caberoy, Brenda A. Schulman, Frank Sicheri, Mike Tyers, Gary Kleiger Dec 2019

Robust Cullin-Ring Ligase Function Is Established By A Multiplicity Of Poly-Ubiquitylation Pathways, Spencer Hill, Kurt Reichermeier, Daniel C. Scott, Lorena Samentar, Jasmin Coulombe-Huntington, Luisa Izzi, Xiaojing Tang, Rebeca Ibarra, Thierry Bertomeu, Annie Moridian, Michael J. Sweredoski, Nora Caberoy, Brenda A. Schulman, Frank Sicheri, Mike Tyers, Gary Kleiger

Life Sciences Faculty Research

The cullin-RING ligases (CRLs) form the major family of E3 ubiquitin ligases. The prototypic CRLs in yeast, called SCF enzymes, employ a single E2 enzyme, Cdc34, to build poly-ubiquitin chains required for degradation. In contrast, six different human E2 and E3 enzyme activities, including Cdc34 orthologs UBE2R1 and UBE2R2, appear to mediate SCF-catalyzed substrate polyubiquitylation in vitro. The combinatorial interplay of these enzymes raises questions about genetic buffering of SCFs in human cells and challenges the dogma that E3s alone determine substrate specificity. To enable the quantitative comparisons of SCF-dependent ubiquitylation reactions with physiological enzyme concentrations, mass spectrometry was employed …


Nanomaterials For Biosensing Lipopolysaccharide, Palak Sondhi, Helal Maruf, Keith Stine Dec 2019

Nanomaterials For Biosensing Lipopolysaccharide, Palak Sondhi, Helal Maruf, Keith Stine

Chemistry & Biochemistry Faculty Works

Lipopolysaccharides (LPS) are endotoxins, hazardous and toxic inflammatory stimulators released from the outer membrane of Gram-negative bacteria, and are the major cause of septic shock giving rise to millions of fatal illnesses worldwide. There is an urgent need to identify and detect these molecules selectively and rapidly. Pathogen detection has been done by traditional as well as biosensor-based methods. Nanomaterial based biosensors can assist in achieving these goals and have tremendous potential. The biosensing techniques developed are low-cost, easy to operate, and give a fast response. Due to extremely small size, large surface area, and scope for surface modification, nanomaterials …


Transcription Regulation Of Human Il1b Gene In Monocytes And Lymphoid Cd4 T Cells, Sree H. Pulugulla Dec 2019

Transcription Regulation Of Human Il1b Gene In Monocytes And Lymphoid Cd4 T Cells, Sree H. Pulugulla

Electronic Theses and Dissertations

Cytokines are key regulators of the inflammatory response and play an important role in facilitating intercellular communication between various immune cell types. Interleukin‑1β (IL‑1β) is a potent pro-inflammatory cytokine that is required for robust initiation of innate immune response and subsequent development of adaptive immunity. IL-1β is first synthesized as an inactive cytoplasmic, non‑glycosylated, precursor molecule (proIL‑1β) by monocytes and macrophages in response to invading pathogenic microbes. The activation of caspase‑1 by inflammasomes cleaves proIL-1β into mature biologically active IL-1β that is released from cells via a non-classical, endoplasmic reticulum‑independent secretory pathway directly from the cytoplasm via Gasdermin D membrane …


Delivery Of Small Molecule And Rna Using Synthetic Polymeric Micelles And Multifunctional Exosomes For The Treatment Of Type 1 Diabetes, Yang Peng Dec 2019

Delivery Of Small Molecule And Rna Using Synthetic Polymeric Micelles And Multifunctional Exosomes For The Treatment Of Type 1 Diabetes, Yang Peng

Theses & Dissertations

Type 1 diabetes is one of the most challenging chronic autoimmune diseases. The destruction and dysfunction of insulin-secreting β cells are the results of inflammatory infiltration and the synergistic effect of multiple immune cells. The aim of this dissertation is to develop novel and reliable therapeutic approaches to advance the treatment of T1D: including chemical modification of a broad-spectrum immunosuppressant, co-application of small molecule based immune intervention and siRNA based β cell preservative therapy, and administration of a PI3K-δ/γ dual inhibitor to specifically target immune cells, utilizing synthetic polymeric micelles or natural produced multi-functional exosomes derived from human bone marrow …


Cholesterol Biosynthesis In The Nervous System With An Emphasis On Desmosterolosis, Luke Allen Dec 2019

Cholesterol Biosynthesis In The Nervous System With An Emphasis On Desmosterolosis, Luke Allen

Theses & Dissertations

Cholesterol biosynthesis is integral to proper neurodevelopment due to the reliance on de novo synthesis of cholesterol in the brain. Disruptions in this process have devastating outcomes for human life characterized by several phenotypic manifestations concomitant with developmental delay. The cholesterol biosynthesis disorder desmosterolosis is an extremely rare disorder with a severe clinical phenotype, however, the models used to study this disease are not well characterized. In addition to genetic disruptions in cholesterol biosynthesis, pharmacological perturbation is an understudied side effect of many commonly prescribed drugs. Here we present a characterization of the sterol profile of the mouse model of …


The Role Of Reactive Oxygen Species In Regulating Macrophage And Fibroblast Activation Within The Breast Cancer Tumor Microenvironment, Brandon J. Griess Dec 2019

The Role Of Reactive Oxygen Species In Regulating Macrophage And Fibroblast Activation Within The Breast Cancer Tumor Microenvironment, Brandon J. Griess

Theses & Dissertations

The tumor microenvironment (TME) is a key determining factor in breast cancer, especially the more aggressive subtype triple negative breast cancer (TNBC). The activated fibroblasts and macrophages within the TME have many tumor promoting functions. Therefore, targeting their activation presents a novel therapeutic approach in TNBC. My work studied the role of reactive oxygen species (ROS) during fibroblast and macrophage activation in breast cancer.

My studies showed that expression of the secreted antioxidant enzyme, EcSOD, is silenced in breast cancer samples, in part, via increased promoter methylation. The re-expression of EcSOD inhibited c-Met activation in the TNBC cell line, MDA-MB231. …


Defining The Role Of Tyrosine Phosphorylation In The Regulation Of Connexin43 In Cardiac Diseases, Li Zheng Dec 2019

Defining The Role Of Tyrosine Phosphorylation In The Regulation Of Connexin43 In Cardiac Diseases, Li Zheng

Theses & Dissertations

Connexins are integral membrane proteins that oligomerize to form gap junction channels. Ions and small molecules diffuse intercellularly through these channels, allowing individual cellular events to synchronize into the functional response of an entire organ. Gap junction channels composed of Connexin43 (Cx43) mediate electrical coupling and impulse propagation in the normal working myocardium. In the failing heart, Cx43 remodeling (decreased expression, altered phosphorylation state, loss at intercalated discs, and increased presence at lateral membranes) contributes to rhythm disturbances and contractile dysfunction. While there is considerable information regarding key interactions of Cx43 in the regulation of gap junction channels, unfortunately, the …


Alanyl-Trna Synthetase Quality Control Prevents Global Dysregulation Of The Escherichia Coli Proteome, Paul Kelly, Nicholas Backes, Kyle Mohler, Christopher Buser, Arundhati Kavoor, Jesse Rinehart, Gregory Phillips, Michael Ibba Dec 2019

Alanyl-Trna Synthetase Quality Control Prevents Global Dysregulation Of The Escherichia Coli Proteome, Paul Kelly, Nicholas Backes, Kyle Mohler, Christopher Buser, Arundhati Kavoor, Jesse Rinehart, Gregory Phillips, Michael Ibba

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Mechanisms have evolved to prevent errors in replication, transcription, and translation of genetic material, with translational errors occurring most frequently. Errors in protein synthesis can occur at two steps, during tRNA aminoacylation and ribosome decoding. Recent advances in protein mass spectrometry have indicated that previous reports of translational errors have potentially underestimated the frequency of these events, but also that the majority of translational errors occur during ribosomal decoding, suggesting that aminoacylation errors are evolutionarily less tolerated. Despite that interpretation, there is evidence that some aminoacylation errors may be regulated, and thus provide a benefit to the cell, while others …


Uncovering New Mechanisms Of Cdc34 And Cullin-Ring Activity, Spencer Hill Dec 2019

Uncovering New Mechanisms Of Cdc34 And Cullin-Ring Activity, Spencer Hill

UNLV Theses, Dissertations, Professional Papers, and Capstones

Ubiquitylation is a cellular regulatory system found in all eukaryotic cells, which has managed to find a role in most pathways imaginable. The system works fundamentally through the ligation of a small protein known as ubiquitin onto a substrate. Depending on the context of the ubiquitin ligation, the substrate can be directed towards a number of cellular fates, the best-studied being degradation of the substrate. While originally thought of as a signal for cellular disposal units to degrade aberrant proteins, we now know that ubiquitin plays a highly nuanced role in cellular epistasis, controlling everything from the cell cycle to …


The Perplexing Paradox Of Clostridioides (Clostridium) Difficile Infection (Cdi) - Analysis Of Anti-Germinants As Part Of Cdi Prophylaxis, Christopher Yip Dec 2019

The Perplexing Paradox Of Clostridioides (Clostridium) Difficile Infection (Cdi) - Analysis Of Anti-Germinants As Part Of Cdi Prophylaxis, Christopher Yip

UNLV Theses, Dissertations, Professional Papers, and Capstones

Clostridioides (Clostridium) difficile infections (CDI) have become the leading cause of nosocomial antibiotic-associated diarrhea worldwide. Under normal circumstances, bacteria found in the gastrointestinal tract provide a barrier against C. difficile colonization. Upon antibiotic therapy, the protective barrier is lost as the microbial community becomes depleted thus providing the opportunity for C. difficile to colonize the human gut. Exposure to taurocholate, a bile acid produced within the mammalian gastrointestinal tract, causes C. difficile spores to begin their transition, a process known as germination, from metabolically dormant structures to toxin-producing cells. As germination is required for the onset of CDI, anti-germination compounds …


Oh Chemistry Of Non-Methane Organic Gases (Nmogs) Emitted From Laboratory And Ambient Biomass Burning Smoke: Evaluating The Influence Of Furans And Oxygenated Aromatics On Ozone And Secondary Nmog Formation, Matthew M. Coggon, Christopher Y. Lim, Abigail R. Koss, Kanako Sekimoto, Bin Yuan, Jessica B. Gilman, David H. Hagan, Vanessa Selimovic, Kyle J. Zarzana, Steven S. Brown, James M Roberts, Markus Müller, Robert Yokelson, Armin Wisthaler, Jordan E. Krechmer, Jose L. Jimenez, Christopher Cappa, Jesse H. Kroll, Joost De Gouw, Carsten Warneke Dec 2019

Oh Chemistry Of Non-Methane Organic Gases (Nmogs) Emitted From Laboratory And Ambient Biomass Burning Smoke: Evaluating The Influence Of Furans And Oxygenated Aromatics On Ozone And Secondary Nmog Formation, Matthew M. Coggon, Christopher Y. Lim, Abigail R. Koss, Kanako Sekimoto, Bin Yuan, Jessica B. Gilman, David H. Hagan, Vanessa Selimovic, Kyle J. Zarzana, Steven S. Brown, James M Roberts, Markus Müller, Robert Yokelson, Armin Wisthaler, Jordan E. Krechmer, Jose L. Jimenez, Christopher Cappa, Jesse H. Kroll, Joost De Gouw, Carsten Warneke

Chemistry and Biochemistry Faculty Publications

Chamber oxidation experiments conducted at the Fire Sciences Laboratory in 2016 are evaluated to identify important chemical processes contributing to the hydroxy radical (OH) chemistry of biomass burning non-methane organic gases (NMOGs). Based on the decay of primary carbon measured by proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS), it is confirmed that furans and oxygenated aromatics are among the NMOGs emitted from western United States fuel types with the highest reactivities towards OH. The oxidation processes and formation of secondary NMOG masses measured by PTR-ToF-MS and iodide-clustering time-of-flight chemical ionization mass spectrometry (I-CIMS) is interpreted using a box model employing …


Characterization Of The Nuclear Pore Complex In Red Alga, Cyanidioschyzon Merolae, Michelle Veronin Dec 2019

Characterization Of The Nuclear Pore Complex In Red Alga, Cyanidioschyzon Merolae, Michelle Veronin

Health and Kinesiology Theses

Cyanidioschyzon merolae (C. merolae) is a primitive, unicellular species of red alga that is considered to be one of the simplest self-sustaining eukaryotes. The highly elementary nature of C. merolae makes it an excellent model organism for studying evolution as well as cell function and organelle communication. In our study, we hypothesize that C. merolae contains the minimal assembly of proteins to make up their Nuclear Pore Complexes (NPCs), and hence are the first ancestral NPCs. NPCs are essential for basic nuclear transport in the cell. They are embedded in the double membrane of the nucleus, the …


Total Phenolic And Flavonoid Contents Of Anacyclus Pyrethrum Link Plant Extracts And Their Antioxidant Activity, Hanane Elazzouzi, Nadia Zekri, Touriya Zair, Mohamed Alaoui El Belghiti Dec 2019

Total Phenolic And Flavonoid Contents Of Anacyclus Pyrethrum Link Plant Extracts And Their Antioxidant Activity, Hanane Elazzouzi, Nadia Zekri, Touriya Zair, Mohamed Alaoui El Belghiti

Karbala International Journal of Modern Science

In this study, Anacyclus pyrethrm Link (roots) is a medicinal plant from the Asteraceae family, endemic to Morocco and is widely used in traditional medicine; was investigated for their total phenolics, flavonoids contents and antioxidant activities. The quantitative estimation of total flavonoids and phenols by the colorimetric method showed that the extracts were prepared by fractionation of the crude hydromethanolic extract (ethyl acetate and n-butanol). The phenol content of various extracts, estimated by the Folin-Ciocalteu method, was between 1,44 and 12,82 mg EAG / g extract of A. pyrethrum. Moreover, it is clear that ethyl acetate was the …


Structural Basis Of Non-Canonical Transcriptional Regulation By The A-Bound Iron-Sulfur Protein Whib1 In M. Tuberculosis, Tao Wan, Shanren Li, Daisy Guiza Beltran, Andrew Schacht, Lu Zhang, Donald F. Becker, Limei Zhang Dec 2019

Structural Basis Of Non-Canonical Transcriptional Regulation By The A-Bound Iron-Sulfur Protein Whib1 In M. Tuberculosis, Tao Wan, Shanren Li, Daisy Guiza Beltran, Andrew Schacht, Lu Zhang, Donald F. Becker, Limei Zhang

Department of Biochemistry: Faculty Publications

WhiB1 is amonomeric iron–sulfur cluster-containing transcription factor in the WhiB-like family that is widely distributed in actinobacteria including the notoriously persistent pathogen Mycobacterium tuberculosis (M. tuberculosis). WhiB1 plays multiple roles in regulating cell growth and responding to nitric oxide stress inM. tuberculosis, but its underlying mechanism is unclear. Here we report a 1.85 A° -resolution crystal structure of the [4Fe–4S] cluster-bound (holo- )WhiB1 in complex with the C-terminal domain of the 70-family primary sigma factor A of M. tuberculosis containing the conserved region 4 (A 4). Region 4 of the 70-family primary sigma factors is commonly used by transcription factors …


Determination Of Optimal Mild Organic Solvents Of Pdnps For Carbon-Carbon Coupling Reactions, Trina Biswas Dec 2019

Determination Of Optimal Mild Organic Solvents Of Pdnps For Carbon-Carbon Coupling Reactions, Trina Biswas

Honors College Theses

The synthesis of nanoparticles is an increasingly popular field of interest. The application of nanoparticles is especially popular in the field of nanocatalysts. Metal nanoparticles (NPs) are favorable for catalysis because of the large surface area to volume ratio, which allows them to catalyze a variety of reactions using lesser amounts of active material. As the field of nanoparticle research expands, efforts are being made to create more sustainable approaches to the synthesis of these particles. This research sought to translate the benefits of peptide-based synthesis to organic solvents and use less toxic organic solvents for carbon-carbon coupling reactions, like …


Simultaneous Delivery Of Crispr/Cas Complexes And Donor Dna Using Cell-Penetrating Peptide-Adaptors, Maria Mugure Mbugua Dec 2019

Simultaneous Delivery Of Crispr/Cas Complexes And Donor Dna Using Cell-Penetrating Peptide-Adaptors, Maria Mugure Mbugua

Master of Science in Chemical Sciences Theses

Clustered Regularly Interspaced Palindromic Repeats (CRISPR)/CRISPR associated protein (Cas) complexes are becoming the preferred technique for genomic editing because of their precision, efficiency and accuracy. Prokaryotes naturally use CRISPR/Cas complexes to detect and eliminate foreign nucleic acids after infections. Cas proteins form complexes with RNA to cause double-stranded breaks (DSBs) in the invader’s sequence. Researchers are trying to use CRISPR complexes to treat many human diseases caused by these mutations. Research using CRISPR/Cas systems in the treatments of many of the genetic diseases has resulted in misfortunes (such as toxicity to the patient or new diseases) because of the method …


High-Throughput Nano-Scale Characterization Of Membrane Proteins Using Fluorescence-Detection Size-Exclusion Chromatography, Alex J. Vecchio, Robert M. Stroud Dec 2019

High-Throughput Nano-Scale Characterization Of Membrane Proteins Using Fluorescence-Detection Size-Exclusion Chromatography, Alex J. Vecchio, Robert M. Stroud

Department of Biochemistry: Faculty Publications

Structural biology has revealed predicting heterologous expression levels, homogeneity, and stability of a protein from its primary structure are exceedingly difficult. Membrane proteins, in particular, present numerous challenges that make obtaining milligram quantities of quality samples problematic. For structural and functional investigation of these molecules, however, this is what is required. Fluorescence size-exclusion chromatography (F-SEC), a technique where a protein of biological interest is fused to green fluorescent protein (GFP) and monitored, circumvents many bottlenecks inherent to membrane protein structural biology. In vivo expression yields, as well as in vitro homogeneity and stability, can be rapidly evaluated utilizing nanogram quantities …


Alzheimer's And Amyloid Beta: Amyloidogenicity And Tauopathy Via Dyshomeostatic Interactions Of Amyloid Beta, Jordan Tillinghast Dec 2019

Alzheimer's And Amyloid Beta: Amyloidogenicity And Tauopathy Via Dyshomeostatic Interactions Of Amyloid Beta, Jordan Tillinghast

Senior Honors Theses

This paper reviews functions of Amyloid-β (Aβ) in healthy individuals compared to the consequences of aberrant Aβ in Alzheimer’s disease (AD). As extraneuronal Aβ accumulation and plaque formation are characteristics of AD, it is reasonable to infer a pivotal role for Aβ in AD pathogenesis. Establishing progress of the disease as well as the mechanism of neurodegeneration from AD have proven difficult (Selkoe, 1994). This thesis provides evidence suggesting the pathogenesis of AD is due to dysfunctional neuronal processes involving Aβ’s synaptic malfunction, abnormal interaction with tau, and disruption of neuronal homeostasis. Significant evidence demonstrates that AD symptoms are partially …


Differential Analysis Of Individual Centruroides Vittatus Venom Dec 2019

Differential Analysis Of Individual Centruroides Vittatus Venom

Symposium of Student Scholars

Scorpions are well known, venomous arthropods (Class: Arachnida, Order: Scorpiones). Their venom is composed of neurotoxins, proteases and cytotoxic peptides which plays a major role in toxic components such as inflammatory, antimicrobial and hemolytic activity. Venom is key for the survival of scorpions as it is involved in defense against prey and in feeding on predators. Centruroides vittatus (bark scorpion) is commonly found in North America, with habitat ranges from Nebraska to southern Texas. For this study, scorpions were harvested from Laredo, Texas (27˚57’ N, 99˚43’ W), held in captivity for 4 months and fed a consistent diet of Acheta …


Multidrug Resistance Regulators Mara, Soxs, Rob, And Rama Repress Flagellar Gene Expression And Motility In Salmonella Enterica Serovar Typhimurium, Srinivas Thota, Lon Chubiz Dec 2019

Multidrug Resistance Regulators Mara, Soxs, Rob, And Rama Repress Flagellar Gene Expression And Motility In Salmonella Enterica Serovar Typhimurium, Srinivas Thota, Lon Chubiz

Biology Department Faculty Works

Production of flagella is costly and subject to global multilayered regulation, which is reflected in the hierarchical control of flagellar production in many bacterial species. For Salmonella enterica serovar Typhimurium and its relatives, global regulation of flagellar production primarily occurs through the control of flhDC transcription and mRNA translation. In this study, the roles of the homologous multidrug resistance regulators MarA, SoxS, Rob, and RamA (constituting the mar-sox-rob regulon in S. Typhimurium) in regulating flagellar gene expression were explored. Each of these regulators was found to inhibit flagellar gene expression, production of flagella, and motility. To different degrees, repression via …


Dual Activities Of Plant Cgmp-Dependent Protein Kinase And Its Roles In Gibberellin Signaling And Salt Stress., Qingwen Shen, Xinqiao Zhan, Pei Yang, Jing Li, Jie Chen, Bing Tang, Xuemin Wang, Xuemin Wang, Yueyun Hong Dec 2019

Dual Activities Of Plant Cgmp-Dependent Protein Kinase And Its Roles In Gibberellin Signaling And Salt Stress., Qingwen Shen, Xinqiao Zhan, Pei Yang, Jing Li, Jie Chen, Bing Tang, Xuemin Wang, Xuemin Wang, Yueyun Hong

Biology Department Faculty Works

Cyclic GMP (cGMP) is an important regulator in eukaryotes, and cGMP-dependent protein kinase (PKG) plays a key role in perceiving cellular cGMP in diverse physiological processes in animals. However, the molecular identity, property, and function of PKG in plants remain elusive. In this study, we have identified PKG from plants and characterized its role in mediating the gibberellin (GA) response in rice (Oryza sativa). PKGs from plants are structurally unique with an additional type 2C protein phosphatase domain. Rice PKG possesses both protein kinase and phosphatase activities, and cGMP stimulates its kinase activity but inhibits its phosphatase activity. One of …


Characterization Of The Dimerization Domains On The Mannose-6-Phosphate/Insulin-Like Growth Factor Ii Receptor, Tyler Degener Dec 2019

Characterization Of The Dimerization Domains On The Mannose-6-Phosphate/Insulin-Like Growth Factor Ii Receptor, Tyler Degener

Theses/Capstones/Creative Projects

The mannose-6-phosphate/insulin-like growth factor II (M6P/IGF2) receptor is a transmembrane protein known to sequester growth factors from the extracellular matrix. This behavior suggests a mechanism of tumor suppression. Structurally, the receptor’s extracellular region is segmented into 15 homologous repeats, which are divided further into 5 triplet domains, labelled 1-3, 4-6, 7-9, 10-12, and 13-15. What is notable about the triplets is their propensity to form dimers with triplets on a second M6P/IGF2 receptor. In fact, previous studies indicate that this protein functions optimally when dimerized. Thus, the purpose of this experiment is to characterize these domain interactions. Using a urea …