Nytarsha Brown Houston, TX Texas Southern University Junior Chemistry Faculty Advisor: Irene C. Solomon, Associate Professor Department of Physiology and Biophysics
COMPARISON OF CONNEXIIN mRNA IN BRAINSTEM VERSUS BRAIN OF C57BL/6 MICE NYTARSHA BROWN(1), IRENE C. SOLOMON(2) Gap junctions, special channels whose pores allow a variety of substances to diffuse between the cytoplasm of the neurons, function in intercellular communication. Connexins (Cx) are structural proteins associated with these gap junctions. There are five abundantly expressed connexins (Cx26, Cx30, Cx32, Cx36, Cx43) in the mammalian central nervous system (CNS). We will try to determine, not only, whether differences in the expression of Cxs in brainstem versus brain (cortex) exist but also whether various isoforms are similarly expressed in brainstem and brain. Reverse Transcription- Polymerase Chain Reaction (RT-PCR) will be used on tissue RNA samples obtained from brainstem and brain of C57 BL/6 mice using primers specific for each connexin isoform. Agrose gel electrophoresis will then be used in order to visualize Cx mRNA expression, which will be quantified by densitometric analysis on the ethiduim bromide stained gel. Image acquisition using a Kodak Image Station 440 is then used to capture the image of the stained gel. The findings may not only help evaluate the similarities and differences in the connexins but may someday aid in discovering other roles that these connexins play in the Central Nervous System. (1)Texas Southern University, 3100 Cleburne Street, Houston, TX 77004 (2)Department of Physiology and Biophysics, University at Stony Brook , Stony Brook, NY 11794-8661
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	Enixy Collado Mercado Caguas, Puerto Rico Stony Brook University First Year Master Marine Sciences Faculty Advisor: Anne McElroy, Associate Professor Marine Sciences Research Center
IMMUNOLOGICAL RESPONSE OF MUMMICHOG FISH (Fundulus heteroclitus) TO PESTICIDES ENIXY COLLADO MERCADO, ANNE MCELROY, ROBIN BARNES AND LOURDES MENA Phagocytosis is the first line of defense against invading bodies in vertebrates. Macrophages and neutrophils are the main cell types responsible for phagocytosis. Phagocytosis and others hematological, nonspecific defensive assays are used in lobster and commercial important fish species (carps and salmonids) but are less often applied to fishes from mashes area. The goal will be develop a method for monitor health of immune response in Mummichog fish (Fudulus heteroclitus) that is derived from techniques used in lobsters, salmons and humans. This assay is an additional part of on going study of acute toxicity and sublethal effects of helicopter mosquito spraying in salt marshes. Pesticides used will be methropene (larvacide) and resmethrin (adulticide) where cage fish and shrimps will be use as a model of salt marsh organisms. Experiments will be performed in natural environment (grass marshes) and we expect lower phagocytic index levels in fish exposed to both pesticides in comparison to reference fish. The lower capacity of the fish to engulf foreign particles would make the fish more susceptible or vulnerable to diseases. It is of great importance to determine if the wild populations of fish are being stressed and immunosuppressed by the pesticides sprayed on Long Island. Marine Sciences Research Center, New York Stony Brook, N.Y. 11794-5000
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	Natasha Cover Coram, New York Virginia Union University Junior Biology Faculty Advisor: Joav Prives, Professor Department of Pharmacological Sciences
REGULATION OF SYNAPSE FORMATION IN MUSCLE CELLS NATASHA COVER(1), JOAV PRIVES(2), CHRISTI WESTON(2) AND PETER HALLOCK(2) A critical step during synapse formation is the clustering of neurotransmitter receptors at the postsynaptic membrane. Our lab is using the neuromuscular junction (NMJ) as a model synapse. The NMJ is where the motor neuron terminates and forms synapse on the skeletal muscles cells. Every neuronal action potential that reaches the NMJ leads to the release of the neurotransmitter, acetylcholine (ACh), which diffuses across the synaptic, cleft and activates acetylcholine receptors (AChRs). Communication across this synapse depends on high density of AChRs in the postsynaptic membrane. Our experiments use cultured muscle cells to study the events responsible for the concentration of AChRs under nerve endings. We are investigating the regulatory pathway that couples signaling by the neuronal protein agrin and the extracellular protein laminin to the aggregation of AChRs into high-density clusters. Myasthenia Gravis is an autoimmune disorder in which circulating antibody directed against AChR or other components of the agrin-signaling pathway impairs clustering and produces a deficiency in the density of AChRs in the NMJ. By increasing our understanding of the molecular mechanisms regulating AChR clustering, this project can contribute to future treatment for Myasthenia Gravis. (1)Department of Virginia Union University, 1500 North Lombardy Street, Richmond, VA 23220 (2)Department of Molecular Pharmacology, Stony Brook University, Stony Brook, New York 11794-5140B
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	Huldah Haynes Dover, DE Delaware State University Sophomore Biology Faculty Advisor: Holly Miller, Assistant Professor Department of Pharmacological Sciences
RNA INTERFERENCE HULDAH HAYNES(1) HOLLY MILLER(2) A new discovery of 2002 has scientists all ears. RNA interference (RNAi) is a technique that specifically decreases or ‘knocks down’ the amount of a target protein that a cell makes. Some potential uses for RNAi are to decrease the level of a DNA polymerase (a protein that synthesizes DNA) or a protein that helps cancer cells proliferate. The overall goal of this project is to ‘knock down’ DNA polymerase ι (pol ι) in mammalian cells using RNAi. The cellular function of this protein is not clear but it is believed to play an important role in DNA repair. To ‘knock down’ pol ι in mammalian cells, a foreign plasmid (ring like structure of DNA) must be inserted into the cell. In order to be sure that a cell has taken up the plasmid, a gene for antibiotic resistance (which enables the cells to grow in the presence of antibiotics) is used. Hygromycin (HYG) and neomycin are common antibiotics for mammalian tissue culture. The first step of this work was to create a plasmid that contains the hygromyocin resistance gene (HYG). In this project neomycin resistant mouse cells are used. Because an alternative resistance is required in the plasmid, HYG was chosen. The polymerase chain reaction was used to amplify or copy the antibiotic resistance HYG gene. Agarose gel electrophoresis was used to purify and analyze samples throughout the project. HYG was ligated into a plasmid to generate pMH1-HYG and transformation was used to insert a foreign plasmid into E. coli. Having generated pMH1-HYG, the second step is to insert a short DNA sequence into the HYG plasmid which will direct the ‘knock down’ of pol ι. Reduction of the pol ι should give insight as to how this protein functions in mammals and will also suggest how pol ι may interact with other proteins. These experiments will help answer some questions but undoubtedly lead to more questions regarding pol ι´s role in the cell. (1)Delaware State University, 1200 N. DuPont Hwy. Dover, DE 19901 (2)Department of Pharmacology, University at Stony Brook, Stony Brook, NY, 11794-5000
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	Kheyandra D. Lewis Easton, PA Philadelphia University Junior Biology Faculty Advisor: Hermann Schindelin, Assistant Professor Department of Structural Biology and Biochemistry
SOLVING MUTANT STRUCTURES OF THE BACTERIAL MoeB-MoaD COMPLEX TO PROVIDE INSIGHT FOR THE MECHANISM OF UBIQUITIN ACTIVATION KHEYANDRA D. LEWIS(1), HERMANN SCHINDELIN(2) AND IMSANG LEE(2) Members of the E1 enzyme family activate ubiquitin, a small protein modifier, with a role in multiple processes such as cellular endocytosis, cell autophagy, DNA repair, inflammatory responses, and HIV budding. The method of activation by this particular group of enzymes occurs in two steps: first ATP is hydrolyzed to cause adenylation of the C terminus in ubiquitin, then ubiquitin becomes covalently attached to a conserved cysteine side chain of the E1 enzyme by means of a thioester bond. Sharing a similar mechanism with the E1 enzyme catalyzed reaction is the MoeB catalyzed reaction. MoeB is an Escherichia coli protein, which together in complex with an additional E. coli protein, MoaD, is a part of the evolutionarily conserved pathway known as Molybdenum Cofactor (Moco) Biosynthesis. A comparison of the amino acid sequence, suggests that MoeB has a mechanism of action that closely resembles the first step of the ubiquitin-activating step. Despite close similarity in mechanism and fold, unlike the ubiquitin activation reaction, there is no intermediate thioester bond formation between MoeB and MoaD. Instead, a sulphurtransferase converts MoaD acyl-adenylate to a thiocarboxylate, which in turn acts as the sulphur donor during Moco biosynthesis. These similarities give support to the notion that E1 enzymes and ubiquitin are derived from ancestral genes that are related to both MoeB and MoaD. To view the components of these catalyzed reactions, we will use X-ray crystallography. By using the active sites of mutant complexes of MoeB-MoaD to produce arrays of crystals under varying conditions, one is able to then use X-ray diffraction beams to determine the structure of the complexes. The goal is to find ideal structures from several MoeB-MoaD complexes, whose residues will help to further the understanding of the ubiquitin pathway (1)Department of Biology, Philadelphia University, School House Lane & Henry Avenue, Philadelphia, PA 19144 (2)Department of Structural Biology and Biochemistry, Stony Brook University, N.Y. 11794
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	Leachien Ricks Queens, NY Stony Brook University Junior Mechanical Engineering Faculty Advisor: Yi-Xian Qin, Associate Professor Department of Biomedical Engineering
THE USE OF NANOINDENTATION TO DETERMINE REGIONAL DEPENDENCE OF MATERIAL PROPERTIES IN NEW AND OLD BONE LEACHIEN RICKS, YI-XIAN QIN AND SUZANNE FERRERI Fluid flow in bone that arises from physiologic loading and mechanical loading has been shown to stimulate new bone formation in particular regions. Numerous studies have shown that physiological and mechanical loading induce new bone formation due to the skeleton´s sensitivity to certain types of stimuli. These stimuli vary in strain rate, strain frequency and strain magnitude. Interest in the quality of the new bone formed has been stimulated by knowledge of the effects of the various stimuli. Therefore the objective of this study is to compare the material properties of old and new bone. In addition we will investigate whether or not these properties are regionally dependent. Nanoindentation will be used to assess the material properties and will provide a means to evaluate the quality of this new bone. It will also be used determine if there is a regional dependence of material properties in new and old bone. Our hypothesis is that new bone forms on the outer periosteal and inner endosteal surfaces of bone, at sites that correspond to peak strain gradients. Strain gradients occur when stress, which is force over area, is put on a bone. Bone then responds to this loading by the amount of strain, or deformation, it undergoes. A strong correlation was found between peak strain magnitude and the amount of new bone formed when the bones were subjected to an internal fluid stimulus. It is also hypothesized that new bone will be lower in elastic modulus, a parameter that describes the resistance of a material to deformation due to tension or compression. New bone is also believed to be lower in hardness, a parameter that describes the material resistance to permanent deformation. A lower elastic modulus and hardness will result because new bone is less mineralized than old bone. Lastly, it is hypothesized that material properties will be regionally dependent due to the variability in mineralization of new bone. Knowledge of the microscale properties of newly formed bone will enable scientists in the future to develop and implement biomechanically based interventions to reduce the risk of fracture. Department of Biomedical Engineering, Stony Brook University, Stony Brook, NY 11790
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	Olúwátóbèrù Thomas Richmond, CA San Francisco State University Junior Mechanical Engineering Faculty Advisor: Fu-Pen Chiang, Distinguished Professor Department of Mechanical Engineering
CRACK PROPAGATION IN TiAl AS INVESTIGATED BY SIEM (SPECKLE INTERFEROMETRY WITH ELECTRON MICROSCOPY) OLÚWÁTÓBÈRÙ THOMAS(1) FU-PEN CHIANG(2) AND WANG KAI(2) Metal composites have assumed a prominent role in manufacturing. Titanium Aluminide (TiAl) has gained worldwide attention because of the potential it holds to replace its counterparts in the aerospace, automobile, and energy industries. Because of its increased stiffness, superior temperature strengths, improved wear resistance, and near-isotropic behavior, TiAl will inevitably become one of the most important building materials of the future. The purpose of this study is to investigate the propagation of cracks in a specimen of TiAl. Before a way can be found to stop the growth of cracks across the sample, the fracture mechanisms must first be observed and the propagation of cracks analyzed. With SIEM, the growth of a crack in the TiAl sample can be studied readily and accurately. Experimental analysis shows that changes in grain orientation and alignment within the boundaries of separate grain colonies contribute to fracture resistance. (1)School of Engineering, San Francisco State University, 1600 Holloway Avenue, San Francisco, CA, 94132 (2)Department of Mechanical Engineering, Stony Brook University, N.Y. 11794
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	Sandra Tinta Bronx, NY Lehman College CUNY First Year Masters Computer Science Faculty Advisor: Y. Annie Liu, Associate Professor Department of Computer Science
SCHEMA-BASED DATA ACQUISITION FOR WEB APPLICATIONS SANDRA TINTA(1) AND Y. ANNIE LIU(2) Increasingly more applications are developed for the web, and most of these applications require the user to enter data. The kinds of data required by the application drive the implementation of the user interface. Consequently, for every web application that has a different set of requirements on data, a new user interface will have to be developed. Most data can be described using XML Schema, which is a language for describing vocabularies of data and specifying constraints on data. Generating web user interfaces automatically from XML Schema will greatly reduce the time spent in the implementation of user interfaces. The generated user interface can check that the entered data satisfy the constraints on data, since these constraints are also specified in XML Schema. Automatic generation of user interfaces will prove useful for rapid prototyping, even though some customization may be required for special applications. This project surveys existing schema-based tools for manipulating data and seeks to develop a prototype for such a tool that can run on a web server and that combines the advantages of existing tools. (1)Lehman College of The City University of New York, 250 Bedford Park Boulevard West, Bronx, NY 10468 (2)Department of Computer Science Engineering, Stony Brook University, Stony Brook, N.Y. 11794
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	Jason White St. Louis, MO Xavier University of LA Senior Physics Faculty Advisor: Alex Doboli, Assistant Professor Department of Electrical Engineering
MOVING TOWARDS CIRCUIT DESIGN BEYOND THE THIRD DIMENSION JASON WHITE(1), ALEX DOBOLI(2) AND HUI ZHANG(2) An analog circuit is an electrical circuit that provides a continuous relationship/ variable signal between its input and output. Without analog circuits, the creation of new technological innovations would not be possible in devices such as cell phones, palm pilots, and navigation systems. Analog circuits are a big part of the microelectronics industry and as electronic devices become smaller, the designs become more complex. In order to visualize the many possibilities for designing microelectronic systems, a simulation is used. The simulation of an analog circuit allows electrical engineers to synthesize and test a circuit to see if it works without wasting the time and money necessary for building an analog circuit. In order to have a simulation program that engineers can use, we are developing a graphical user interface (GUI)/computer aided design (CAD) interface. Graphical user interfaces and computer aided design interfaces are tools that make interacting with a computer easier by allowing the user to manipulate data in order to display graphical images and draw schematics. GUIs and CADs are helpful, however, since humans only have vision in three dimensions the graphical images and schematics are displayed, at most, in three dimensions. Being able to display information in more than three dimensions would allow many parameters of analog circuits to be checked at one time rather than three parameters at a time and allow for more designs. The enigma becomes how can we display or create multiple dimensions beyond the third dimension? The goal of this research is to create a GUI/ CAD tool that can display multiple dimensions which will create more possibilities for the synthesis of analog circuits in the microelectronics industry. (1)Department of Physics, Xavier University, 1 Drexel Dr., New Orleans, LA 70125 (2)Department of Electrical and Computer Engineering, Stony Brook University, N.Y. 11794
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	Brian D. Williams Killeen, TX Savannah State University Senior Biology Faculty Advisor: Stella Tsirka, Assistant Professor Department of Pharmacological Sciences
PURIFICATION AND TESTING OF THE CHEMOTACTIC POTENCY OF MCP-1 BRIAN D. WILLIAMS(1), STELLA E. TSIRKA(2) AND JOHN J. SHEEHAN(2) Excitotoxicity is a key component in many neurodegenerative diseases, including Alzheimer´s disease, ischemic stroke, and multiple sclerosis. Tissue plasminogen activator (tPA) plays important roles in the brain after excitotoxic injury. It is released by both neurons and microglia, and mediates neuronal death and microglial activation. tPA is an extracellular serine protease that cleaves the zymogen plasminogen into plasmin. Chemokines are a superfamily of small-secreted proteins that function in recruiting immunocells such as microglia. We hypothesize that in mice plasmin directly cleaves the chemokine monocyte chemoattractant protein-1 (MCP-1), and this cleavage increases the potency of the chemotactic gradient that recruits microglia to sites of neuronal injury. Microglia are specialized macrophages that react to neurons of the central nervous system (CNS). When there is neuronal damage, the microglia will invade the region and secrete growth factors. In the case of neuronal death, the microglia will surround the dead tissue and remove it by the process of phagocytosis. Excessive recruitment of microglia can sometimes prove harmful, especially when MCP-1 becomes more active. In doing this study, we will investigate if plasmin cleaves MCP-1 and makes it more active allowing the recruitment of more microglia. Our results will help us to better understand the mechanisms of microglia and their recruitment by MCP-1. (1) Department of Biology and Life Sciences, Savannah State University, 3219 College St., Savannah, GA 31404 (2) Department and Program of Pharmacological Sciences , Stony Brook University, N.Y. 11794-8651
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	Oladapo Yeku Queens, NY Medgar Evers (CUNY) Sophomore Biology Faculty Advisor: Guangwei Du and Michael Frohman Department of Molecular Pharmacology
TARGETED PROTEIN DEGRADATION IN MAMMALIAN CELLS OLADAPO YEKU(1) GUANGWEI DU(2) MICHAEL FROHMAN(2) Loss of function analysis at the DNA (gene knockout) and RNA (RNA interference) level has been successfully used to analyze gene function in plant and animal cells. Here we describe an attempt to establish a technique to silence expression at the protein level in mammalian cells, based on a recent demonstration that controlled redirection of target proteins to the proteasome is sufficient for their degradation in yeast. A protein subunit known as FRB was fused to a target protein (Green fluorescent protein). The complementary subunit FKBP was fused to a subunit of the proteasome complex. A rapamycin analogue was then added to trigger dimerization of the FRB and FKBP subunits, thus promoting translocation of the FRB-tagged GFP to the proteasome. In theory, close proximity of the GFP to the proteasome complex should result in its degradation. Loss of GFP was assessed quantitatively over various time periods using western blotting and immunofluorescence. This method, if successful, could provide a powerful tool for the study of gene and protein function. (1)Medgar Evers College, Brooklyn, New York 11225 (2)Department of Molecular Pharmacology, Stony Brook University, Stony Brook, New York 11794-5140