Biochemistry, Biophysics, and Structural Biology Commons^™

Synthesis, Characterization, And Biological Activity Of Imidazolium Salts, David Weader

Williams Honors College, Honors Research Projects

Nonmuscle invasive bladder cancer (NMIBC) inflicts thousands of Americans annually, and is typically treated with the immunotherapy BCG. However, due to a BCG shortage, there is a new need for novel treatments of NMIBC. Addressing this issue, several imidazolium salt derivatives were synthesized and characterized with the intent of treatment within the bladder. These imidazolium salts were tested against different human bladder cancer cell lines in vitro to determine their reactivity and cytotoxicity. Among these results are GI50 concentrations for each drug, which is the concentration of drug needed to see growth inhibition in 50% of treated cells. Previous published …

Insights Into The Structure, Pharmacology, And Evolution Of The Glycine Transporter 2, Ashley Bryan Lopez

Open Access Theses & Dissertations

The balance between neuronal excitation and inhibition in the central nervous system is vital for survival. Overexcitation can be toxic and lead to certain manias or even death. On the other hand, extremely low levels of inhibition cause hyperekplexia, pain, and even forms of autism. Neuronal inhibition is, for the most part, achieved by two neurotransmitters, GABA and glycine. Levels of both neurotransmitters in the synaptic cleft are meticulously regulated by the GABA and glycine transporters, respectively, which belong to the solute carrier 6 (SLC6) family of neurotransmitter transporters and share structural similarities with other family members. In Chapter 1, …

Molecular And Functional Characterization Of Α-Galactosyl Epitopes In Trypanosoma Cruzi, Uriel Ortega Rodriguez

Open Access Theses & Dissertations

Trypanosoma cruzi, the causative agent of Chagas disease (CD) currently affects 6-7 million people across the world. Currently, only two drugs, benznidazole and nifurtimox, are available for treatment of CD and they are highly toxic and less effective in the chronic stage of the disease. Specific biomarkers for diagnosis and follow-up of treatment do not exist in the clinical settings. following chemotherapy, patients take approximately 10-20 years to exhibit negative seroconversion with the conventional serology assays. Moreover, there is no vaccine available to prevent or treat CD. T. cruzi contains a complex cell surface consisting of several classes of glycoconjugates …

Expression, Purification, And Characterization Of Recombinant Human Npas2, Brenda Moreno

Open Access Theses & Dissertations

Many living organisms have biological clocks known as circadian rhythms that control various physiological and behavioral processes, tailored with the day and night. At the cellular level, the circadian clock drives daily rhythms with a transcriptional-translational feedback loop (TTFL) established by multiple transcription factors and genes under their regulation. Circadian Locomotor Output Cycles Kaput (CLOCK) and Brain and Muscle ARNT-like protein 1 (BMAL1) are two of those transcription factors, initiating the TTFL by forming a heterodimeric complex that bind to DNA promoters. Neuronal PAS domain protein 2 (NPAS2) is a functional analog to CLOCK. Less knowledge about NPAS2, in contrast …

Antioxidant Intervention In Radical-Induced Protein Damage, Emmanuel Zubia

Open Access Theses & Dissertations

Many environmental toxicants are known to increase free-radical production, which can create cellular havoc and lead to a myriad of ailments. Antioxidants can be used to ameliorate the prevalence of free radicals, potentially serving as prophylactics against radical-induced disorders. Many exogenous antioxidants are phytochemicals in origin, such as ascorbic acid, gallic acid, and ellagic acid. Some of these antioxidants can be found in traditional medicine, like tanshinone and brazilin, which have been studied and are known to have low cytotoxicity. Here we test the radical rescue capacity of a selected group of phytochemical antioxidants on protein rescue models. Specifically, our …

Plant-Soil Interactions Dominate Soil Microbial Respiration And Soil Organic Carbon Sequestration In A Subtropical Moist Evergreen Broadleaved Forest In China, Zhijie Yang

Theses and Dissertations--Plant and Soil Sciences

Tropical forest soils contain one-third of global soil carbon (C). The warm and moist climate in tropical forests leads to rapid soil organic carbon (SOC) decomposition, with the highest soil microbial respiration rates in the world, so even a slight change in soil C and microbial respiration could affect atmospheric carbon dioxide concentration. However, there remains a lack of understanding of the mechanisms driving microbial respiration in tropical forests, due to different climate and biophysical drivers compared to temperate or boreal forests. Furthermore, forest conversions (from natural forests to plantations) are most widespread in tropical regions, leading to a loss …

Development Of Assessment Tools Of Package/Product Systems For A Sustainable Food Chain, Kompal Joshi

Doctoral

Post-harvest life of fresh produce is limited due to high metabolic activity and microbial spoilage. Modified atmosphere packaging (MAP) has proven to be one of the most effective techniques to extend the shelf life of fresh produce commercially. Obtaining of an optimum concentration of oxygen and carbon dioxide inside the package depends upon the product properties, the environmental conditions of the cold chain, the permeable film, some of which are subjected to natural variability during the food distribution chain. This variability may generate produce that is out of specification that will lead to food waste. Uncertainty analysis of this problem …

Termination-Independent Role Of Rat1 In Cotranscriptional Splicing In Budding Yeast, Zuzer Hakimuddin Dhoondia

Wayne State University Dissertations

Rat1 is a 5′→3′ exoribonuclease in budding yeast belonging to the XRN-family of nucleases. It is a highly conserved protein with homologs being present in fission yeast, flies, worms, mice and humans. Rat1 and its homolog in metazoan have been shown to function in multiple facets of RNA metabolism. In this study, we report a novel role of Rat1 in splicing of pre-mRNA in budding yeast. In the absence of the functional Rat1 in the nucleus, an increase in the level of unspliced transcripts was observed in yeast cells. Strand-specific TRO analysis revealed that the accumulation of unspliced transcripts upon …

Novel Insights Into The Critical Role Of Cardiolipin In Cellular Metabolism And Mitochondrial Physiology, Jiajia Ji

Wayne State University Dissertations

Cardiolipin (CL) is the signature phospholipid of mitochondria. CL and its remodeling exert critical roles in biological processes both inside and outside of mitochondria. CL abnormalities have been associated with various mitochondrial disorders and aging. Understanding the role of CL in mitochondrial physiology and cellular metabolism could provide valuable insights into cell biology and human health. Several metabolic alterations have been reported in CL-deficient cells, including accumulated lactate, decreased PDH activity, and decreased TCA cycle function. This dissertation connected these findings by showing abnormal NAD+ metabolism in various models lacking CL. Importantly, it shows that NAD+ supplementation improves mitochondrial function …

Germinal Center B Cell Expression Of Aire Regulates Antibody Diversification And Autoimmunity, Jordan Zheng Zhou

Wayne State University Dissertations

B cells are a unique subset of immune cells that, in response to antigen, diversify their antibody repertoire to generate progressively higher affinity antibodies of different isotypes through the processes of affinity maturation/somatic hypermutation (SHM) and class switch recombination (CSR). One of the major sites in which this diversification occurs is in T cell dependent microanatomical structures known as germinal centers (GC). Here, we find that GC B cells express the protein, autoimmune regulator (Aire) in a CD40 dependent manner. In these cells, Aire interacts with activation induced cytidine deaminase (AID), the protein responsible for initiating the processes of diversification …

Laser Surface Cleaning As A Novel Approach For Genesis Solar Wind Collectors, Martina Schmeling, I. V. Veryovkin, C. E. Tripa

Chemistry: Faculty Publications and Other Works

A new surface cleaning method utilizing lasers has been evaluated for its suitability to Genesis solar wind collectors

Structural Basis For The Stereoselective Epoxidation Reaction In Polyether Natural Product Biosynthesis, Qian Wang

Open Access Theses & Dissertations

Polycyclic polyether natural products have received much attention due to their diverse biological activities, ranging from extreme toxicity to therapeutic properties, including antimicrobial, antifungal, and anticancer activities. They are featured by multiple cyclic ether groups that vary in the number, size, and arrangement. Ionophore polyethers contain multiple tetrahydrofuran and tetrahydropyran rings connected by either spiroketal systems or carbon-carbon single bonds. Over the past 40 years, significant progress has been made in deciphering polyether biosynThesis pathways. Remarkably, all members of the polycyclic polyether family are thought to be generated via a common biosynthetic scheme. In 1983, Cane, Celmer, and Westly proposed …

A Role For Mek3 In The Oncogenesis Of Acute Lymphocytic Leukemia: Inactivation Of Mapk P38 Promotes Cell Proliferation Through Enhanced Degradation Of Mutant Mek3, Yoshira Marie Ayala-Marin

Open Access Theses & Dissertations

In the United States, pediatric leukemia has the second-highest cancer mortality rate in Hispanic children, especially Acute Lymphocytic Leukemia (ALL). While treatment of ALL has improved overall the five-year survival rate of ~90 %, not everyone has benefited. Twenty percent of them will experience relapse, and from these, 30 - 50 % will die. Unfortunately, the cause behind these dreadful statistics is poorly understood due to the complex etiology of this disease. Thereby, it is essential to identify potential oncogenic proteins that promote ALL so that new strategies can be developed to diagnose and treat this cancer. Whole Exome Sequencing …

Non-Invasive Method For Leptin Supplementation In Zebrafish (Danio Rerio), Regan Mcnamara

Williams Honors College, Honors Research Projects

I tested the hypothesis that recombinant leptin protein can be introduced to zebrafish in vivo through non-invasive soaking in a solution containing the protein. One way to study various molecules’ effects in vivo is through intraperitoneal or intracerebroventricular injections during the embryonic or larval stage, which is invasive, difficult to administer, and can have a high mortality rate. 48 hours post fertilization (hpf) zebrafish were soaked in a His-tagged recombinant leptin protein solution at 10 nM and 100 nM concentrations (produced by Genscript). After soaking, zebrafish larvae were washed extensively to remove all recombinant protein on their exterior before homogenization. …

Pneumovirus Infections: Understanding Rsv And Hmpv Entry, Replication, And Spread, Jonathan T. Kinder

Theses and Dissertations--Molecular and Cellular Biochemistry

Pneumoviruses including human metapneumovirus (HMPV) and respiratory syncytial virus (RSV) are significant causes of respiratory tract infections globally. Children, elderly, and immunocompromised patients are at the greatest risk for developing severe infections, which can have devastating outcomes. Although these viruses are ubiquitous with significant impacts on human health, there are no antivirals or vaccines available. The only FDA approved therapy is a monoclonal antibody for RSV, given prophylactically during the infectious season, and this treatment is only available for high risk infants. The work presented in this thesis aims to increase our understanding of how these viruses enter, replicate, and …

Eecp Improves Markers Of Functional Capacity Regardless Of Underlying Ranolazine Therapy, Sanaz Ziad, Jamil Malik, Obinna Isiguzo, Lang Xu, Leqi Chen, Annette Cox, Sachin Shah

School of Pharmacy Faculty Articles

OBJECTIVE: Enhanced external counterpulsation (EECP) and ranolazine are approved treatments for patients with chronic stable angina by the United States Food and Drug Administration (FDA). Whether EECP offers clinical benefits regardless of underlying ranolazine therapy needs further investigation.

METHODS: This was a retrospective evaluation of patients referred to a specialized EECP center. Patients having data on 6-Minute Walk Distance (6MWD) or Duke Activity Status Index (DASI) were categorized into two groups (EECP with ranolazine or EECP only). The primary endpoints were change in 6MWD and DASI before and after a full course of EECP within each of the two groups. …

Modulation Of K V 7 Channel Deactivation By Pi(4,5)P 2, Carlos A. Villalba-Galea

School of Pharmacy Faculty Articles

The activity of K_V7 channels critically contributes to the regulation of cellular electrical excitability in many cell types. In the central nervous system, the heteromeric K_V7.2/K_V7.3 channel is thought to be the chief molecular entity giving rise to M-currents. These K⁺-currents as so called because they are inhibited by the activation of Gq protein-coupled muscarinic receptors. In general, activation of Gq protein-coupled receptors (GqPCRs) decreases the concentration of the phosphoinositide PI(4,5)P₂ which is required for K_V7 channel activity. It has been recently reported that the deactivation rate of K …

Alternative Splicing Of Mdm4 In Human Melanomas, Abdullah Salem S. Alatawi

Browse all Theses and Dissertations

Melanoma is a potentially lethal type of skin cancer and regarded to be the third most common type of skin cancer. Although melanoma is not as common as basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), it is more likely to metastasize than BCC and SCC. Interestingly, the incidence of melanoma continues to go up (expected 2% in 2020), but the deaths continue to decrease (-5.3% in 2020) due to improvements in detection and treatment. The treatment of melanoma depends on several aspects but most importantly the tumor's stage and the location. In the early stages, melanoma can be …

Development Of A Quantitative Pcr (Qpcr) Based Method For Studying Temporal Dna Degradation In Waterlogged Bone, Isis Thornton

Master of Science in Forensic Science Directed Research Projects

Human activities are often centered around the presence of water, thus it is not surprising that there are many water-related human deaths. Accumulated degree days (ADD), and other aquatic variables may affect DNA retrieval from waterlogged bone. Calcium and collagen in bone can inhibit the PCR necessary to produce an STR profile; the current solution is a time-consuming organic extraction. While there are examples of research on DNA degradation in terrestrial bone over time, there has been little work done on submerged bone samples and they are usually limited to case studies. The major aim of this study was to …

The Oncogenic Role Of The Protein Tyrosine Phosphatase 4a3 (Ptp4a3 Or Prl-3) In T-Cell Acute Lymphoblastic Leukemia, Min Wei

Theses and Dissertations--Molecular and Cellular Biochemistry

T-cell Acute Lymphoblastic Leukemia (T-ALL) is an aggressive blood cancer. There are no immunotherapies and few molecularly targeted therapeutics available for the treatment of this malignancy. The identification and characterization of genes and pathways that drive T-ALL progression is critical for the development of new therapies for T-ALL. The protein tyrosine phosphatase PTP4A3, also known as PRL-3, has been extensively reported to play a causative role in numerous cancers, including several types of blood malignancies. However, its role in T-ALL is not well defined.

Here, we determined that the PRL-3 plays a critical role in T-ALL initiation and progression by …

Epigenetic Implications In Inorganic Arsenic-Mediated Carcinogenesis, Meredith Eckstein

Theses and Dissertations--Molecular and Cellular Biochemistry

Chronic, low dose exposure to inorganic arsenic (iAs) is a public health concern throughout the world, contributing to the development of many diseases, including lung cancer. Several mechanisms for iAs-mediated carcinogenesis have been proposed, of which the production of reactive oxygen species and formation of chromosomal aberrations are the most studied. Another equally important, yet less studied mechanism is dysregulation of epigenetic marks. “Epigenetics” refers to changes that occur on the DNA and chromatin that do not alter base pair identity, but alter compaction, expression, and regulation of specific DNA sequences. There are several types of epigenetic marks including histone …

Uterine Torsion Dystocia Complicated By Perosomus Elumbis In An Angus Calf Associated With A Consanguineous Mating, Alyssa B. Helms, Riley E. Thompson, Sam Lawton, Jessica L. Petersen, Allison Watson, Mee-Ja Sula, David J. Steffen, Brian K. Whitlock

School of Veterinary and Biomedical Sciences: Faculty Publications

A six-year-old multiparous Angus cow was presented for dystocia. Vaginal and rectal examinations revealed an approximately 360° counterclockwise uterine torsion. The torsion was corrected by rolling the cow counterclockwise (three episodes) with the aid of a plank coupled with manual detorsion via the vagina. The placement of obstetric chains followed by manual traction ultimately delivered a stillborn male calf with evidence of vertebral aplasia, arthrogryposis, and abdominal organ herniation. Patient history and subsequent parentage verification revealed that the calf was the result of a consanguineous (mother to son) mating. Tissue samples from the affected calf and blood samples from the …

Identification Of Immunogenic Epitopes That Permit The Detection Of Antigen-Specific T Cell Responses In Multiple Serotypes Of Group B Coxsackievirus Infections, Ninaad Lasrado, Arunakumar Gangaplara, Rajkumar Arumugam, Chandirasegara Massilamany, Sayli Pokal, Yuzhen Zhou, Shi-Hua Xiang, David Steffen, Jay Reddy

School of Veterinary and Biomedical Sciences: Faculty Publications

Coxsackievirus group B (CVB) contains six serotypes that can affect various organs. Some of these organ-specific diseases such as myocarditis and pancreatitis can be caused by more than one serotype. Thus, development of immunological tools common to multiple serotypes is desired. This is especially critical for analyzing antigen-specific T cell responses at a single cell level. To this end, we made efforts to identify the immunogenic epitopes of CVB3 leading us to localize three T cell epitopes within the viral protein 1 (VP1) namely, VP1 681–700, VP1 721–740 and VP1 771–790. First, we confirmed their immunogenicity in the immunization settings. …

First Records Of Established Populations Of Ixodes Scapularis (Acari: Ixodidae) Collected From Three Nebraska Counties, Lindsey E. Nielsen, Roberto Cortinas, Paul D. Fey, Peter C. Iwen, David H. Nielsen

School of Veterinary and Biomedical Sciences: Faculty Publications

Reported cases of Lyme disease in Nebraska have been assumed to be imported from other endemic areas. Previous surveillance efforts provided no evidence of established populations as only individual specimens of Ixodes scapularis (Say) had been collected. In the winter of 2018, adult I. scapularis were found on a dog at Two Rivers State Recreation Area, Douglas County, prompting tick collection at the site and nearby natural areas. In May 2019, all life stages of host-seeking I. scapularis were collected using dragging and flagging techniques in sites located near the Platte River in Douglas, Sarpy, and Saunders counties. This is …

Methods Of Treating Or Preventing Zika Virus Infection, Shi-Hua Xiang, Nicholas Palermo, Asit K. Pattnaik, Aryamay Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

This document relates to methods and materials for treating a mammal having a Zika virus (ZIKV) infection. For example, a composition including one or more non-nucleoside RNA polymerase inhibitors can be administered to a mammal having, or at risk of developing, a ZIKV infection to treat the mammal.

Recent Advances In The Diagnosis Of Classical Swine Fever And Future Perspectives, Lihua Wang, Rachel Madera, Yuzhen Li, D. Scott Mcvey, Jishu Shi

School of Veterinary and Biomedical Sciences: Faculty Publications

Classical swine fever (CSF) is a highly contagious viral disease of pigs, including wild boar. It is regarded as one of the major problems in the pig industry as it is still endemic in many regions of the world and has the potential to cause devastating epidemics, particularly in countries free of the disease. Rapid and reliable diagnosis is of utmost importance in the control of CSF. Since clinical presentations of CSF are highly variable and may be confused with other viral diseases in pigs, laboratory diagnosis is indispensable for an unambiguous diagnosis. On an international level, well-established diagnostic tests …

Evaluation Of A Baculovirus-Expressed Vp2 Subunit Vaccine For The Protection Of White-Tailed Deer (Odocoileus Virginianus) From Epizootic Hemorrhagic Disease, Sun Young Sunwoo, Leela E. Noronha, Igor Morozov, Jessie D. Trujilo, In Joong Kim, Erin E. Schirtzinger, Bonto Faburary, Barbara S. Drolet, Kinga Urbaniak, D. Scott Mcvey, David A. Meekins, Mitchell V. Palmer, Velmurugan Balaraman, William C. Wilson, Juergen A. Richt

School of Veterinary and Biomedical Sciences: Faculty Publications

Epizootic hemorrhagic disease virus (EHDV) is an arthropod-transmitted RNA virus and the causative agent of epizootic hemorrhagic disease (EHD) in wild and domestic ruminants. In North America, white-tailed deer (WTD) experience the highest EHD-related morbidity and mortality, although clinical disease is reported in cattle during severe epizootics. No commercially licensed EHDV vaccine is available in North America. The objective of this study was to develop and evaluate a subunit vaccine candidate to control EHD in WTD. Recombinant VP2 (rVP2) outer capsid proteins of EHDV serotypes 2 (EHDV-2) and 6 (EHDV-6) were produced in a baculovirus-expression system. Mice and cattle vaccinated …

Amphiphilic Cyclobutenes And Cylobutanes, Wantanee Sittiwong, Patrick H. Dussault, Raul G. Barletta, Robert Powers

School of Veterinary and Biomedical Sciences: Faculty Publications

This disclosure relates to amphiphilic compounds containing a cyclobutene or cyclobutane moiety. In some embodiments, the compounds are useful for treating infection by Mycobacterium such as Mycobacterium tuberculosis. Cyclobutene containing compounds are also useful as monomers in the preparation of amphiphilic polymers.

Non-Neuroinvasivevirusesand Uses Thereof, Gregory A. Smith, Patricia J. Sollars, Gary E. Pickard, Ekaterina E. Heldwein

School of Veterinary and Biomedical Sciences: Faculty Publications

Provided herein are compositions and methods for vaccination and research applications. In particular, provided herein are non-neuroinvasive herpesviruses and alpha herpesviruses and uses thereof.

Mitochondrial Metabolism In Astrocytes Regulates Brain Bioenergetics, Neurotransmission And Redox Balance, Jordan Rose, Christian Brian, Aglaia Pappa, Mihalis I. Panayiotidi, Rodrigo Franco

School of Veterinary and Biomedical Sciences: Faculty Publications

In the brain, mitochondrial metabolism has been largely associated with energy production, and its dysfunction is linked to neuronal cell loss. However, the functional role of mitochondria in glial cells has been poorly studied. Recent reports have demonstrated unequivocally that astrocytes do not require mitochondria to meet their bioenergetics demands. Then, the question remaining is, what is the functional role of mitochondria in astrocytes? In this work, we review current evidence demonstrating that mitochondrial central carbon metabolism in astrocytes regulates overall brain bioenergetics, neurotransmitter homeostasis and redox balance. Emphasis is placed in detailing carbon source utilization (glucose and fatty acids), …