Current Search:  biotechnology (x)

View All Items

  • CSV Spreadsheet
(1 - 5 of 5)
The Role of Type-I Interferon in Limiting Spread and Killing of an Oncolytic RNA Virus in Prostate Cells
Title
The Role of Type-I Interferon in Limiting Spread and Killing of an Oncolytic RNA Virus in Prostate Cells.
Creator
Kedarinath, Kritika, Parks, Griffith, Chakrabarti, Ratna, Altomare, Deborah, University of Central Florida
Abstract / Description
Prostate cancer is the second most prevalent cancer amongst men and there is an urgent need to address viable therapeutic options for its treatment. Development of viruses which target and kill cancer cells has gained momentum due to the first FDA approved oncolytic virus for treating human cancer patients. Our previous work with the RNA virus, Parainfluenza Virus 5 (PIV5), has led to the generation of mutants that are potential candidates for oncolytic viruses: 1) the hyperfusogenic (P/V/F)...
Show moreProstate cancer is the second most prevalent cancer amongst men and there is an urgent need to address viable therapeutic options for its treatment. Development of viruses which target and kill cancer cells has gained momentum due to the first FDA approved oncolytic virus for treating human cancer patients. Our previous work with the RNA virus, Parainfluenza Virus 5 (PIV5), has led to the generation of mutants that are potential candidates for oncolytic viruses: 1) the hyperfusogenic (P/V/F) mutant has a mutated P/V and fusion gene which activates anti-viral responses and causes massive cell-cell fusion respectively, and 2) the Leader mutant has a mutated viral genomic promoter which kills cells due to overactive viral gene expression. The P/V/F mutant has shown effectiveness in reducing prostate tumor burden in a mouse model system, however, the specificity of these viruses is unclear, i.e. targeting cancerous prostate cells while leaving uninvolved cells unaffected. In this study, we addressed how these PIV5 mutants replicate in and killed tumor versus benign human prostate cells. Flow cytometry demonstrated that the mutants are able to infect and replicate in prostate tumor cells (22Rv1), resulting in effective cell killing. However, these mutants showed highly restricted spread in benign prostatic hyperplasia cells (BPH-1). Upon further exploration, it was determined that the restriction observed in the BPH-1 cells is due to the induction and signaling of type-I Interferon (IFN). This was confirmed upon treatment with an IFN-? neutralizing antibody, which relieved restricted spread of mutants in benign cells. BPH-1 cells infected with the mutants also showed upregulation of key anti-viral, IFN-induced genes such as TLR3, IFIT1, and OAS2. Upon characterization of the mutant viruses in an additional metastatic prostate cancer cell line (C4-2B), a restriction in viral spread was observed. The restricted spread did not correlate with production of high levels of type-I IFN, suggesting that other cytokines or intracellular factors can limit replication in tumor cells. Therefore, these studies lay the groundwork for further improving the specificity of oncolytic PIV5 mutants by exploiting type-I IFN pathways as well as other anti-viral factors.
Show less
Date Issued
2016
Identifier
CFE0006468, ucf:51445
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0006468
ENGINEERING A NEW FORM OF ENCLOSURE: INTERNATIONAL CONVERGENCE IN GMO REGULATION
Title
ENGINEERING A NEW FORM OF ENCLOSURE: INTERNATIONAL CONVERGENCE IN GMO REGULATION.
Creator
Altif, Jessica, Jacques, Peter, University of Central Florida
Abstract / Description
As society begins to recognize its impact on ecological systems, the belief that modern political institutions can offer a sense of control and certainty, as well as protect the health of its citizens, is increasingly questioned. In an era of uncertainty, faith in science and technology to alleviate industrial impacts on the environment is often embraced by policymakers yet questioned by the public who see the authoritative role of the sciences in the political sphere as contributing to...
Show moreAs society begins to recognize its impact on ecological systems, the belief that modern political institutions can offer a sense of control and certainty, as well as protect the health of its citizens, is increasingly questioned. In an era of uncertainty, faith in science and technology to alleviate industrial impacts on the environment is often embraced by policymakers yet questioned by the public who see the authoritative role of the sciences in the political sphere as contributing to global risk. The development of biotechnology, specifically genetically modified food, places an anthropocentric focus on resolving and/or adapting to environmental degradation, further reflecting an adherence to the dominant social paradigm to address the consequences of modernization. In order to explicate the dualism of human/nature relations inherent in biotechnology, the focus of this research provides an exploration into two competing paradigms of genetically modified organism (GMO) regulatory policy: scientific rationality and social rationality. Through a careful examination of the evolution of GMO regulation in the United States and the European Union, the precarious relationships between science and politics and progress and precaution reveal an actual convergence instead of divergence between these two actors in the international system. Although existing literature proclaims a division between the values and ethics of U.S. and EU environmental policy, the end result of this comparison in GMO regulation illustrates that in both the risk assessment and precautionary approaches, nature is still viewed as an instrument for advancing enclosure of the commons.
Show less
Date Issued
2010
Identifier
CFE0003021, ucf:48371
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0003021
AMERICAN AGRIBUSINESS AND BIOTECHNOLOGY: A NEW ERA OF FARMING
Title
AMERICAN AGRIBUSINESS AND BIOTECHNOLOGY: A NEW ERA OF FARMING.
Creator
Ryan, Nicole M, Sadri, Houman, University of Central Florida
Abstract / Description
In the past fifty years there has been an incredible amount of change made to the agrarian system of the United States. New discoveries in the realm of biotechnology led to the adoption of genetically modified organisms (GMOs) in agriculture, and transformed the industry. Due to regulatory policies set during the nineteen-eighties this technology was able to benefit from widespread commercialization. Today, we see the effects of this approach and are entering into a highly volatile political...
Show moreIn the past fifty years there has been an incredible amount of change made to the agrarian system of the United States. New discoveries in the realm of biotechnology led to the adoption of genetically modified organisms (GMOs) in agriculture, and transformed the industry. Due to regulatory policies set during the nineteen-eighties this technology was able to benefit from widespread commercialization. Today, we see the effects of this approach and are entering into a highly volatile political climate in regard to GMOs. This paper aims to provide an analysis of the regulatory system in place and the discrepancies that exist in US policy. The factors evaluated through this thesis include the current US regulatory approach, advancements in biotechnology, and a comparative perspective on US and EU systems. In each of these reviews it is also relevant to mention consumer opinion on GMOs and the role of interest groups. It is important for every American consumer to understand the politics and technology behind their meals. Through the analysis of recent judicial decisions and the enactment of new laws this thesis explains how the use of GMOs in agriculture is causing an unprecedented change to the political structures in place.
Show less
Date Issued
2016
Identifier
CFH2000035, ucf:45586
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFH2000035
LOW COST PRODUCTION OF PROINSULIN IN TOBACCO AND LETTUCE CHLOROPLASTS FOR INJECTABLE OR ORAL DELIVERY OF FUNCTIONAL INSULIN AND C-PEPTIDE
Title
LOW COST PRODUCTION OF PROINSULIN IN TOBACCO AND LETTUCE CHLOROPLASTS FOR INJECTABLE OR ORAL DELIVERY OF FUNCTIONAL INSULIN AND C-PEPTIDE.
Creator
Burberry, Diane, Daniell, Henry, University of Central Florida
Abstract / Description
Current treatment for type I diabetes includes delivery of insulin via injection or pump, which is highly invasive and expensive. The production of chloroplast-derived proinsulin should reduce cost and facilitate oral delivery. Therefore, tobacco and lettuce chloroplasts were transformed with the cholera toxin B subunit fused with human proinsulin (A, B, and C peptides) containing three furin cleavage sites (CTB-PFx3). Transplastomic lines were confirmed for site-specific integration of...
Show moreCurrent treatment for type I diabetes includes delivery of insulin via injection or pump, which is highly invasive and expensive. The production of chloroplast-derived proinsulin should reduce cost and facilitate oral delivery. Therefore, tobacco and lettuce chloroplasts were transformed with the cholera toxin B subunit fused with human proinsulin (A, B, and C peptides) containing three furin cleavage sites (CTB-PFx3). Transplastomic lines were confirmed for site-specific integration of transgene and homoplasmy. Old tobacco leaves accumulated proinsulin up to 47% of total leaf protein (TLP). Old lettuce leaves accumulated proinsulin up to 53% TLP. Accumulation was so stable that up to ~40% proinsulin in TLP was observed even in senescent and dried lettuce leaves, facilitating their processing and storage in the field. Based on the yield of only monomers and dimers of proinsulin (3 mg/g leaf, a significant underestimation), with a 50% loss of protein during the purification process, one acre of tobacco could yield up to 20 million daily doses of insulin per year. Proinsulin from tobacco leaves was purified up to 98% using metal affinity chromatography without any His-tag. Furin protease cleaved insulin peptides in vitro. Oral delivery of unprocessed proinsulin bioencapsulated in plant cells or injectable delivery into mice showed reduction in blood glucose levels similar to processed commercial insulin. C-peptide should aid in longterm treatment of diabetic complications including stimulation of nerve and renal functions. Hyper-expression of functional proinsulin and exceptional stability in dehydrated leaves offer a low cost platform for oral and injectable delivery of cleavable proinsulin.
Show less
Date Issued
2010
Identifier
CFE0003257, ucf:48554
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0003257
Expression of Lipase from Mycobacterium tuberculosis in Nicotiana tobacum and Lactuca sativa Chloroplasts
Title
Expression of Lipase from Mycobacterium tuberculosis in Nicotiana tobacum and Lactuca sativa Chloroplasts.
Creator
Lloyd, Bethany, Daniell, Henry, Kolattukudy, Pappachan, Self, William, University of Central Florida
Abstract / Description
Tuberculosis (TB), caused by the bacterium Mycobacterium tuberculosis (M. tuberculosis), is a global threat and the leading cause of death among individuals infected with HIV. TB treatment requires multi-drug cocktails, due to the increasing rates of drug resistance of the bacterium. With multi-drug cocktails, strains have been documented to be resistant to all major drugs in the fight against TB. Since the strains are drug resistant, it calls for an increasing need for vaccine and treatment...
Show moreTuberculosis (TB), caused by the bacterium Mycobacterium tuberculosis (M. tuberculosis), is a global threat and the leading cause of death among individuals infected with HIV. TB treatment requires multi-drug cocktails, due to the increasing rates of drug resistance of the bacterium. With multi-drug cocktails, strains have been documented to be resistant to all major drugs in the fight against TB. Since the strains are drug resistant, it calls for an increasing need for vaccine and treatment development for the purpose of preventing and managing the disease. The most widely distributed vaccine against TB is Bacillus Calmette-Gue(&)#180;rin (BCG). Apart from being ineffective in certain individuals, BCG offers only a limited timeframe of protection, is unable to serve as a booster for extending this timeframe and due to the intradermal route of administration requires costly refrigeration and syringes.LipY protein, a M. tuberculosis cell wall lipase, may play a potential role as not only a drug target but a potential vaccine antigen. LipY is known to be up-regulated during both active infection and dormancy. In a previous study, sera from TB patients had shown an IgG and IgM response against it. In this study transplastomic Lactuca sativa and Nicotiana tabacum plants were generated by transforming the chloroplasts through the particle delivery system with pLsDv-LipY and pLD-LipY vectors respectively. The vectors were flanked by the native trnI and trnA gene sequence to facilitate homologous recombination into the chloroplast genome. The vector also contained the 16S rRNA promoter, the selectable marker gene, aadA for specitinomycin resistance, the rbcL untranslated region, the LsPpsbA (PpsbA in N. tabacum) promoter, and LsTpsbA (tpsbA in N. tabacum) untranslated region. Site specific integration of the LipY gene into the chloroplast genome was confirmed by PCR. Homoplasmy of transplastomic plants was confirmed by Southern blot analysis. These plants showed normal growth and were fertile, producing seeds. Once germinated, these seeds did not show Mendelian segregation of the transgene. Immunoblot analysis was performed to analyze the expression of the LipY protein. A 40kDa protein was produced in E.coli, and a 25kDa protein was produced in chloroplasts; a cleaved product in chloroplasts is still valuable as an antigen for vaccine production. Future studies will include testing this chloroplast derived antigen in animal models for vaccine development. ?
Show less
Date Issued
2012
Identifier
CFE0004502, ucf:49289
Format
Document (PDF)
PURL
http://purl.flvc.org/ucf/fd/CFE0004502