Lital Asher, M.A.

Apoptosis and Autoimmune Disease    abstract
Advisor:  Amer Beg

Benjamin J. Brewer, M.A.

A New Way to Test for the Presence of Lvh Genes in Legionellae    abstract
Advisor: Ron Prywes

David M. Kelly, M.A.

Intrabodies: Design, Delivery and Application    abstract
Advisor: Chloe Bulinski

Anna Kirman, M.A.

Accurate Pre- morbid Detection of Alzheimer’s Disease Using Monoclonal Antibody Technology    abstract
Advisor:  Carol Lin

Agnes Marie Kozak, M.A.

Creating a New Focus for Biotechnology    abstract
Advisor:  Carol Lin

Erika Jeannine Medynski, M.A.

Genomics and Drug Discovery   abstract
Advisor:  Carol Lin

Usman Nazma, M.A.

Gene Therapy: A probable Cure for Breast Cancer    abstract
Advisor:  Lili Yamasaki

Shiri Nitzan, M.A.

The Y- box Protein, Yipsilon Schachtel, and its Role in Oogenesis and Embryonic Development in Drosophila melanogaster    abstract
Advisor:  Tulle Hazelrigg

Melina Rincon, M.A.

Bacteriophage Lytic Enzymes and their Potential Uses as an Anti-bacterial Agent    abstract
Advisor:  Chloe Bulinski

Julius K. Simonelli, M.A.

Hematopoietic Stem Cell Based Gene Therapy For HIV-1 Targeting CCR5 and CXCR4: Is it Possible to Develop HIV-1 Resistant Lymphocytes?    abstract
dvisor:  Carol Lin

Alexander W. Threlfall, M.A.

The Past, Present and Future of Neuroscience: The role of biotechnology in uncovering the mysteries of neurodegenerative and psychiatric disease    abstract
Advisor:  Carol Lin

Apoptosis and Autoimmune Disease

Lital Asher, M.A. 2001

Apoptosis is a well conserved cell death mechanism that plays a role in the development embryos as well as in homeostasis processes in mature organisms. In the immune system, apoptotic cell death plays a role in shaping the developing lymphocyte repertoire, executing cellular effector responses and regulating the lymphocyte population. It is believed that inappropriate and ill-regulated cell death processes participate in the development of autoimmune diseases. It is believed that death receptors, mainly the CD95 (Fas/Apo-1) system, mediate these ill-regulated apoptotic processes and contribute to the autoimmune phenotype. New strategies for treatment of autoimmune diseases are being developed, based on knowledge regarding apoptotic pathways and the regulation of death receptors. Such strategies include development of small molecule therapy and gene therapy for treating autoimmunity.

A New Way to Test for the Presence of Lvh Genes in Legionellae

Benjamin J. Brewer, M.A. 2001

Based on the suggestion of Segal et al., I hypothesized that Lvh genes occur across al Legionella species. Although it is known that Lvh genes exist in all pneumophilous species, their presence in all non-pneumophilous species is unknown. Originally I amplified and sequence four different Lvh genes in five non-pneumophilous species (and in three known pneumophilous control species) with forward and reverse primers to detect the presence of Lvh genes in conserved regions. My sequencing and alignment results were that only four of 24 gene regions of the pneumophilous Legionellae could be sequenced to produce alignments, and only one of 40 gene regions for non-pneumophilous species could be aligned. Although this provided some support for my thesis, it also showed the insensitivity of this methodology and the need for numerous replications.

However, gel electrophoresis showed that the primers amplified four gene regions for all the positive controls and one gene region for the five non-pneumophilous strains. My discovery that primers lose the ability to amplify after three of four days led me to develop a more sensitive detection methodology. With the refined methodology, I tested and analyzed lengths of the conserved regions of all 18 known non-pneumophilous Legionellae. My findings were that one of the Lvh genes existed in 17 of the 18, and two Lvh genes existed in all 18. All the primers for each gene amplified the same length for each gene region for most of the positive controls and for as many of the non-pneumophilous negative controls. Additionally, the gels produced discrete bands across the board, single bands in each lane, indicating a unique region in the whole Legionellae genome. These results are supportive of my hypothesis.

Intrabodies: Design, Delivery and Application

David M. Kelly, M.A. 2001

Intracellularly expressed antibodies, intrabodies, targeting cellular components offer a potent means for treating a plethora of human diseases including cancer, AIDS, and Alzheimer’s disease. The production of these intrabodies is based on phage display technology, which allows highly specific Ab to be isolated. A tremendous variety of intrabody forms can be crafted to act on single of multiple targets from all classes of macromolecules. Gene therapy techniques using both viral and non-viral vectors have been employed in schemes to deliver intrabody genes to selected targets for expression. Below, the design, delivery, and application of intrabodies as therapeutics agents are discussed.

Accurate Pre- morbid Detection of Alzheimer’s Disease Using Monoclonal Antibody Technology

Anna Kirman, M.A. 2001

Alzheimer’s disease (AD) is a progressive, degenerative disease of the brain. It is the most common form of dementia, especially in adults over the age of 65. As medical technologies advance, more adults enjoy a longer life, however they increasingly risk developing AD. Alzheimer’s research has uncovered many mysteries surrounding this devastating disease, however scientists have not yet elucidated the exact etiology. Alois Alzheimer first described the histopathology of the disease in 1906 during autopsy, and the hallmark senile plaques (SP) and neurofibrillary tangles (NFTs) described still remain the basis for accurate diagnosis of AD. Since AD- type dementia resembles symptoms of many other diseases and disorders, current pre- morbid methods of diagnosing AD include a battery of tests that, through a process of elimination, conclude either ‘possible’ AD or ‘probable’ AD. In this paper, I examine Monoclonal Antibody (mAb) technology as a potential approach to enhancing the accuracy of AD diagnosis in living patients. Various in vitro assays using mAbs have been designed to measure biomarkers in biological samples that reflect the disease burden in the brain. However, unless a causal relationship can be established between these biomarkers and disease progression, these tests can only be used in conjunction with those that are routinely administered to further increase precision of the diagnosis. A more convincing strategy to make a faithful pre- morbid diagnosis of AD is to administer radiolabeled mAbs to specifically and selectively label the SPs and subsequently image the brain to locate and quantify the disease burden. This technology has been used for other diseases, including various types of cancers. The application of this technology to brain tumors provides an excellent model for other neurological diseases, especially AD. Obstacles such as the blood brain barrier (BBB) must be circumvented. This paper examines methods of crossing the BBB with the aid of mAbs, and further suggests a drug delivery system that bypasses the BBB in order to transport the labeled mAbs to the brain. Using radiolabeled mAbs as an imaging reagent has the potential to be used as a powerful tool for the pre- morbid and precise diagnosis of AD.

Creating a New Focus for Biotechnology

Agnes Marie Kozak, M.A. 2001

Since the beginning of the explosion of research in genetics, there has been an incredible advance in the technologies used in the laboratory. Biotechnology has pushed the barriers of science further than it could have ever been imagined. This important knowledge and techniques must now be used to expand the knowledge in other areas of research. An incredible application of biotechnology is its use to further the understanding of the human nervous system. Several studies have pioneered this new application for biotechnology, but it is absolutely invaluable that projects like them continue throughout the next century. This research should not only focus on completing the existing knowledge of physiology of the nervous system, but also incorporate the psychological phenomena which occur with in it.

Genomics and Drug Discovery

Erika Jeannine Medynski, M.A. 2001

Prior to genomics, screening, target oriented synthesis, and diversity oriented synthesis were the main approaches used in drug discovery. The field of genomics arose over a century ago, but only recently has it really influenced drug discovery. The field of genomics has made amazing progress and now genomic technologies account for roughly 20% of drug discovery methods. It was not until the 1990’s when genomes began to be sequenced. Sequence comparisons through homology based analysis of microbial genome projects has helped to identify targets. In addition structural genomics, functional genomics, gene expression profiling, and genetic assays has helped to bring new antimicrobial drugs to market. The human genome project has impacted drug discovery in a similar manner. Gene microarrays and bioinformatics have been extremely useful tools in both microbial and human genome analysis, both aiding in target identification and validation. With regard to microarrays, many formats exist, each with the ability to screen large numbers of gene and protein samples. Likewise, databases have been an essential tool to drug discovery, making clustering analysis and homology based comparisons possible. The newest field stemming from genomics is pharmacogenomics, a radical approach to studying gene sequence variations, such as DNA polymorphisms. Pharmacogenomics is expected to significantly influence drug discovery, clinical studies, and to potentially lead to individualized health care.

Gene Therapy: A probable Cure for Breast Cancer

Usman Nazma, M.A. 2001

Breast cancer will be diagnosed in 1 out of every 8 women in the United States in 2001. Although current methods of treatment, such as surgery and chemotherapy, have clinical utility, they are not appropriate for every breast cancer patient. In response to new technologies and a better understanding of the disease, novel therapies are being explored. The intent of this review is to analyze gene therapy as a novel approach to the treatment of breast cancer.

Gene therapy is the introduction of genetic material into a cell in hope of correcting a specific defect. I evaluated the use of five different types of gene therapy in the treatment of mammary carcinomas: they are, gene therapy with the aid of tumor suppressors, gene therapy using Antisense molecules, gene therapy using suicide genes, gene therapy with the aid of the multiple drug resistance gene and the role of gene therapy in cancer vaccines. In addition to these therapies, specific methods of gene introduction or delivery are compared, contrasted and evaluated.

The conclusion of this article is that gene therapy should be explored as a novel treatment of breast cancer. The results are preliminary, but indicate that this novel therapy has the potential to be used consistently on breast cancer patients, regardless of tumor stage or grade. All of the five types of therapies analyzed could have clinical utility. Each type of therapy presented has specific goals, for example gene therapy with the multiple drug resistance gene or suicide gene primarily function to aid current breast cancer treatments, whereas the use of gene therapy in cancer vaccines is considered a novel approach with long term benefits. I believe using gene therapy to produce cancer vaccines will be the prophylactic treatment with the greatest clinical value. The major obstacle that has to be overcome, in order for all gene therapies to succeed, is the development of a better delivery system. The delivery system that show the greatest promise is the "oligoplex".

The Y- box Protein, Yipsilon Schachtel, and its Role in Oogenesis and Embryonic Development in Drosophila melanogaster

Shiri Nitzan, M.A. 2001

Oogenesis and embryonic development in Drosophila melanogaster require the correct spatial and temporal localization of various transcripts in order to progress normally. Various gene products have been demonstrated to play an important role in achieving the timely concentration of gene transcripts required for anteroposterior and dorsoventral axis specification and for the development of specific structures (such as the pole cells that give rise to the adult fly gonads). Many of these gene products have been shown to directly interact with their target transcripts. In my study, I have looked at Yipsilon Schachtel (Yps), a Y- box protein suggested to have a role in pole cells development. Yps has been found associated in a ribonucleoprotein (RNP) complex together with oskar (osk) mRNA which is required for pole cells and abdominal structures specification. It is suggested that through its association with osk mRNA, Yps functions in transporting osk mRNA and in the repression of its translation, antagonizing the function of a second protein 0018- RNA binding (Orb) that acts as osk translational enhancer.

Bacteriophage Lytic Enzymes and their Potential Uses as an Anti-bacterial Agent

Melina Rincon, M.A. 2001

As bacteria become resistant to conventional antibiotics, it is clear that new drugs must be developed in order to control and treat the symptoms of evasive pathogenic strains. Many factors have contributed to the rise in pathogenic strains and modern forms of travel has facilitated the dispersement of these super strains of bacteria that are resistant to most, if not all, forms of currently employed therapy. This phenomenon is further complicated by the fact that some bacterial strains live as carriers within an asymptomatic host, thus spreading to other individuals and serving as a potential source of infection. Thus, as bacteria increasingly acquire resistance, it is imperative that new drug strategies be developed to kill unwanted strains of bacteria, without eliminating or disrupting endogenous beneficent bacteria. One such drug strategy is the use of lytic bacteriophages. Lytic bacteriophages are organisms found in nature that have developed effective systems of killing only those bacterial strains they are capable of infecting. The enzymes encoded by lytic phages weaken the cell wall, thereby leaving a bacterial cell vulnerable to osmotic lysis. For this reason, lytic enzymes derived from lytic bacteriophages can serve as a novel form of therapy that is both specific and lethal to deleterious bacteria.

Hematopoietic Stem Cell Based Gene Therapy

For HIV-1 Targeting CCR5 and CXCR4:

Is it Possible to Develop HIV-1 Resistant Lymphocytes?

Julius K. Simonelli, M.A. 2001

Human immune deficiency virus gains entry into host cells through interactions between envelope glycoproteins and CD4+. This interaction is insufficient to mediate viral entry. Interactions between host cell coreceptors, primarily CCR5 for M-tropic strains and CXCR4 for T-tropic strains, and virion envelope glycoproteins are also required. It has been found that individuals homozygous for a particular mutation within the gene encoding CCR5 are naturally resistant to M-tropic HIV-1 infection. Similarly, individuals homozygous for a mutation in the CXCR4 gene are resistant to T-tropic HIV-1 strains. Interestingly, these phenotypes are not associated with clinical conditions, implicating that the biological function of these chemokine receptors are compensated for by other chemokine receptors with similar functions. These findings provide a foundation for novel, anti-HIV-1, therapeutic approaches aimed toward the reproduction of such phenotypes in human lymphocytes. Both the CCR5 and CXCR4 null phenotypes have been successfully mimicked in-vitro using strategies employing intrakines, intrabodies, ribozymes and group II introns. It is hoped that the successes achieved thus far in-vitro could also be reproduced in-vivo. Ultimately, autologous transplantation using transgenic hematopoietic stem cells that fail to express CCR5 and CXCR4 would allow for the establishment of an HIV-1 resistant cell pool in HIV-1 infected individuals. Subsequently, this pool of cells would be capable of restoring a functional immune system through the normal differentiation and proliferation processes characteristic of hematopoietic stem cells. Numerous difficulties must be overcome, however, before we can expect to achieve this lofty goal.

The Past, Present and Future of Neuroscience:

The role of biotechnology in uncovering the mysteries of neurodegenerative and psychiatric disease

Alexander W. Threlfall, M.A. 2001

During the first half of the 20th century, neuroscience persevered to overcome a universal state of disregard within the scientific community and ultimately emerged as a formidable institution of modern science. Neuroscience’s entry into mainstream science was facilitated by its fusion with several divisions of traditional science. For the next fifty years, modern neuroscience benefited from the incorporation of technology derived from the fields of molecular biology and genetics. Tools such as high-speed gene profiling and transgenic model systems enabled researchers to address many questions concerning neuronal development and function and the roles these traits play in behavior and neurological disease. In addition, the efforts of the Human Genome Project during the 90’s enabled scientists to move closer towards understanding the mechanistic undertones of various neurodegenerative and psychiatric diseases.

As science now enters the ‘Post Genomic or Proteomic Era’, the efforts to understand these enigmatic diseases will be enhanced by the development of functional genomics. The application of this information in the treatment of neurological disease will also depend on the development of novel therapeutic approaches aimed at bypassing the blood brain barrier and targeting the primary sources of disease.

However, the limited ability to accurately model the complexities of neuronal function and human behavior remains a significant obstacle in the progression of research. A complete and comprehensive understanding of the mechanistic underpinnings of neurological disease will depend on another technological breakthrough. This technology must provide a means to observe the molecular complexities of neuronal function in highly polygenic and heterogeneous states of disease. Whether it occurs through the development of novel transgenics, imaging technology, the application of another science (i.e. nanotechnology) or a combination thereof, this transition will likely be the last pivotal event in neuroscience.