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Farnesol-Mediated Regulation Of Hepatic Lipid Metabolism In Heparg Cells, Asmita Pant

Wayne State University Dissertations

Non-alcoholic fatty liver disease is emerging as one of the most common liver disorders worldwide and is characterized by accumulation of triglycerides (TGs) in liver. The endogenous isoprenoid farnesol reduces hepatic TG levels in rodents, and this effect appears to involve at least two nuclear receptors, peroxisome proliferator-activated receptor α and farnesoid X receptor (FXR). However, farnesol’s effects on human hepatic lipid metabolism are currently unknown. The objective of this study is to evaluate how farnesol treatment would affect hepatic lipid accumulation and metabolism in a cellular model of human hepatic steatosis that was created by incubating the hepatocyte-like HepaRG …

The Impact Of Myeloperoxidase And Its Related Oxidants On Metaphase Ii Mouse Oocyte Quality, Faten Nuri Shaeib

Wayne State University Dissertations

Inflammatory reactions mediated by oxidative stress (OS) have been implicated in the deterioration of oocyte quality, which may lead to subfertility. Oxidative stress generated from enhancement of activated macrophages secondary to an inflammatory response are the major source of reactive oxygen species (ROS) such as superoxide (O2•−), hydrogen peroxide (H2O2), hydroxyl radical (•OH), and hypochlorous acid (HOCl), as well as, the pro-inflammatory enzyme myeloperoxidase (MPO). Previously, it has been shown that these ROS have deleterious effect on oocytes; however the link between inflammation through macrophage activity and oocyte quality remains unclear. In this work, we investigated: 1) the mechanism through …

Analyzing Sumo-2/3 Modification In Regulation Of Breast Cancer Progression And Mitotic Chromosome Segregation, Divya Subramonian

Wayne State University Dissertations

Roles of SUMOylation in cancer metastasis and mitotic progression has been elucidated in this thesis. This vital and dynamic post-translational modification has been known to be dysregulated in several kinds of cancer. Global SUMO-2/3 conjugation is upregulated in a mouse metastatic breast cancer cell line, 66cl4, compared to a non-metastatic cell line 168FARN from the same genetic background. In order to identify the SUMO-2/3 substrates, SILAC labeled lysates from the two cell lines were immunopurified using SUMO-2/3 monoclonal antibodies. By using SUMO-2/3 epitope peptide elution (SUMO-2/3-ePIPE) and quantitative mass spectrometry, 66 SUMO-2/3 targets were identified, of which 15 targets are …

Identification Of Lipolysis-Derived Lipid Mediators And The Activation Of A Pro-Inflammatory Cyclooxygenase Pathway, Via Cyclooxygenase-2, In Adipose Tissue, Allison Christine Gartung

Wayne State University Dissertations

Adipose lipolysis triggers pro-inflammatory responses that play critical roles in insulin resistance and associated metabolic syndrome. However, pro-inflammatory mediators generated by adipose lipolysis, particularly in the context of lipid mediators, are poorly defined. In this study, the activation of the beta-3 adrenergic receptor (ADRB3)/hormone sensitive lipase (HSL) pathway, a well-employed model system, was utilized to characterize the pro-inflammatory lipid mediators generated by adipose lipolysis. Cultured adipocytes were treated with an ADRB3 agonist and the media was analyzed for eicosanoids using the LC-MS/MS lipidomic method. Among the characterized eicosanoids, I found that approximately 43 metabolites generated by cyclooxygenase (COX), lipoxygenase, and …

Genome Wide Analysis Identifies Sphingolipid Metabolism As A New Target Of Valproic Acid, Shyamalagauri Jadhav Jadhav

Wayne State University Dissertations

Bipolar disorder (BD), which is characterized by depression and mania, affects about 1% of the total world population. Current treatments are effective in only 40-60% of cases and cause severe side effects. Valproic acid (VPA), a branched short-chain fatty acid, is one of the most widely used drugs for the treatment of BD. Although many hypotheses have been postulated to explain the molecular mechanism of action of this drug in BD, the therapeutic mechanism is not understood. This knowledge gap has hampered the development of new drugs to treat this disorder. To identify candidate pathways affected by VPA, I performed …

Unraveling The Molecular Mechanisms Of Aposematic Pigmentation In Oncopeltus Fasciatus, Jin Liu

Wayne State University Dissertations

Insects display the extraordinary amount of morphological variation, particularly in regard to their colorations. This diversity in coloration exists both in terms of the array of colors as well as the patterning of pigments. Previous studies have revealed that melanin, ommochrome, and pteridine are the most common pigments constituting insect color patterns. Among these three, the melanin pathway has been best studied so far and presents the foundation of the current understanding of insect coloration. At the same time, though, most of this insight relied on work performed in model systems such as Drosophila and Tribolium, which have significant limitations …

Regulation Of Cytochrome C Functions By Phosphorylation, Gargi Mahapatra

Wayne State University Dissertations

The long term goal of my thesis research is to understand how tissue-specific

phosphorylations on the small mitochondrial protein, cytochrome c (Cytc), regulate its

functions, under both physiologically healthy and stressed conditions, and to identify the

cell signaling pathways targeting Cytc. Cytc is a functionally diverse protein that carries

electrons in the electron transport chain and plays a critical role in cellular apoptosis, two

diverse pathways that maintain cellular health that are active under diverse conditions.

Since Cytc plays a pivotal role in both these highly divergent pathways, regulation of the

protein is very important—phosphorylation of the protein under physiological …

Imaging Anti-Proliferative Compounds With Flt-Pet, Christopher Mchugh

Wayne State University Dissertations

Imaging is critical in the detection and management of malignancies, and positron emission tomography (PET) is an imaging approach that provides information regarding cancer physiology through the tracking of molecular pathways and receptors. 3’-fluoro-3’-deoxythymidine (FLT) is a PET tracer designed to image cellular proliferation, which is a hallmark of cancer. FLT has been used to study the response of cancer to a variety of treatments such as chemotherapy, targeted agents, and radiation.

Here we explored FLT retention as a biomarker to monitor the anti-proliferative effect of the synthetic glucocorticoid (GC) dexamethasone (Dex) on non-small cell lung cancer (NSCLC). The basis …

Modeling The Mechanism Underlying Environmental And Genetic Determinants Of Gene Expression And Complex Traits, Gregory Alan Moyerbrailean

Wayne State University Dissertations

Advances in next-generation sequencing technologies and functional genomics strategies have allowed researchers to identify both common and rare genetic variation, to deeply profile gene expression, and even to determine regions of active gene transcription.

While these technologies and strategies have contributed greatly to our understanding of complex traits and diseases, there are many biological questions and analytical issues to be addressed.

Genome-wide association studies (GWAS) have successfully identified large numbers of genetic variants associated with complex traits and diseases. However, in many cases the mechanistic link between the phenotype and associated variant remains unclear. This may be because most variants …

The Role Of Alarmins, Invariant Nkt Cells And Senescence In The Pathophysiology Of Sterile Intra-Amniotic Inflammation, Olesya Plazyo

Wayne State University Dissertations

Preterm birth is defined as the delivery of a live baby prior to the 37th week of gestation. It is the leading cause of neonatal mortality worldwide. Preterm neonates are at a higher risk for short- and long-term complications, and prematurity places significant burden on our society. Elucidation of the mechanisms that lead to spontaneous preterm labor will enable development of therapies to prevent this syndrome. We aimed to study pathological inflammation that is implicated in the pathophysiology of spontaneous preterm labor.

Pathological inflammation can be initiated by the activation of innate immunity either by microorganisms or alarmins, which are …

Role Of Secretory Granule Heterogeneity In Calcium-Triggered Exocytosis, Tejeshwar Rao

Wayne State University Dissertations

The sympathetic nervous system is activated by a variety of threats to organismal homeostasis. The adrenomedullary chromaffin cell is the core effector of sympathetic activity in the peripheral nervous system. By design, the chromaffin cell secretory response is mutable so that release can be rapidly tuned to drive context-dependent changes in physiological function. However, the mechanisms by which this tuning is achieved with such high temporal fidelity and context specificity remain unclear. This represents a major gap in our understanding of the sympatho-adrenal system since it is known to modify the function of nearly every organ system in the body. …

Mechanisms Of Pancreatic Beta Cell Dysfunction In Diabetes, Vaibhav Sidarala

Wayne State University Dissertations

Diabetes is a serious medical condition characterized by decreased insulin secretion from pancreatic β-cells and decreased insulin sensitivity in the peripheral tissues, resulting in elevated levels of blood glucose. According to the International Diabetes Federation, about 387 million cases have been reported worldwide in the year 2013 and it is estimated that about 500 million people would be affected by 2050. Type 2 diabetes, which accounts for about 90% of the total number of cases, is caused by decreased insulin sensitivity in the peripheral tissues and decreased glucose-stimulated insulin secretion from the pancreatic β-cells. The underlying mechanisms involved in β-cell …

Exploration Of Cancer Proliferative Signaling In Chemotherapy Drug Resistance And Mdig-Induced Tumorigenesis, Kai Wu

Wayne State University Dissertations

ABSTRACT

EXPLORATION OF CANCER PROLIFERATIVE SIGNALING IN CHEMOTHERAPY DRUG RESISTANCE AND MDIG-INDUCED TUMORIGENESIS

by

KAI WU

August 2016

Advisor: Dr. Fei Chen

Major: Pharmaceutical Sciences

Degree: Doctor of Philosophy

Aberrant intracellular signaling pathway is one of the major driving forces of malignancy through multiple stages of human cancers. Our study demonstrates that in cancer cells, the signaling pathways are profoundly and actively intertwined with each other so they can synergistically affect cell biology, including promoting development of malignancy and compensating the loss of proliferation or survival signals in responses to anti-tumor drug. Moreover, cancer cells can also adopt “non-canonical” mechanisms …

An Investigation Into The Molecular Basis Underlying Enhancement Of Transcription By The Intron In Budding Yeast, Neha Agarwal Agarwal

Wayne State University Dissertations

It is now quite evident that the introns, which are removed from the primary transcript by the process of splicing, are involved in a variety of important functions in eukaryotic cells. One of the evolutionarily conserved functions of introns is their role in regulating transcription of genes that harbors them. This effect of a splicing-competent intron on transcription is known as ‘Intron-Mediated Enhancement of transcription’ (IME). It has been observed that the intron-containing genes are often transcribed more efficiently than non-intronic genes. However, the molecular mechanism underlying IME in budding yeast and higher eukaryotes is not entirely clear, and that …

Characterization Of The Yeast Cysteine Desulfurase Complex Within The Mitochondrial Fe-S Cluster Biogenesis, Dulmini Pabasara Barupala

Wayne State University Dissertations

Disrupted iron homeostasis within the human body materializes as various disorders. Pathophysiology of many of them relates to iron induced oxidative damage to key cellular components caused by iron accumulation within the tissues. Pertaining to the growing occurrence, cost of patient care and devastating burden associated with these diseases, the call for understanding the role of iron homeostasis within these disorders becomes inevitable. Being an abundant iron containing cofactor, the role of Fe-S clusters in cellular iron homeostasis is indisputable in the case of Friedreich’s ataxia, a disease caused by a deficiency in the protein frataxin that is indispensable during …

Studies Towards Broadening The Substrate Profile And Regulation Of Histone Deacetylase 1, Dhanusha Ashanthi Nalawansha

Wayne State University Dissertations

Aberrant expression of histone deacetylase 1 (HDAC1) is implicated in multiple diseases, including cancer. As a consequence, HDAC1 has emerged as an important therapeutic target for drug development. HDAC1 regulates key cellular processes, such as cell proliferation, apoptosis, and cell survival, by deacetylating both histone and non-histone substrates. Due to the lack of simple tools to identify physiological substrates of HDAC1, the full spectrum of HDAC1 activities in the cell remains unclear. Here, we employed a substrate trapping strategy to identify cellular substrates of HDAC1. Using this approach, we identified mitosis-related protein Eg5 as a substrate. HDAC1 colocalizes with Eg5 …

Cardiolipin Is Required For Optimal Acetyl-Coa Metabolism, Vaishnavi Raja

Wayne State University Dissertations

The phospholipid cardiolipin (CL) is crucial for many cellular functions and signaling pathways, both inside and outside of mitochondria. My thesis focuses on the role of CL in energy metabolism. Many reactions of electron transport and oxidative phosphorylation, the transport of metabolites needed for these processes, and the stabilization of electron transport chain supercomplexes, require CL. Recent studies indicate that CL is required for the synthesis of iron-sulfur (Fe-S) co-factors, which are essential for numerous metabolic pathways. Activation of carnitine-acetylcarnitine translocase, which transports acetyl-CoA into the mitochondria, is CL dependent. The presence of substantial amounts of CL in the peroxisomal …

Loss Of Rap1gap: A Driver In The Progression From Dcis Of The Breast To Idc, Seema Shah

Wayne State University Dissertations

The purpose of this study is to determine the role of Rap1Gap in the progression of DCIS (ductal carcinoma in situ) to IDC (invasive ductal cancer). We employed an in-vitro three-dimensional (3D) overlay model that provides a physiologically relevant microenvironment to study mechanisms of malignant progression. Previous studies from this laboratory aimed to determine specific gene expression changes common to three models of DCIS- MCF10.DCIS, SUM 102 and SUM 225 in comparison to MCF10A cells, a model of non-transformed human mammary epithelium. The expression of 295 genes was found to be significantly altered, with 63 being increased in expression in …