Nine students in Massachusetts' delegation, 24 strong, received awards at Intel ISEF 2013! In addition to the Special Award winners announced last night, this year's Grand Award winners included: Ayush Kumar & Raashed Raziuddin, Advanced Mathematics and Science Academy 3rd place Grand Award - Life Sciences - Biochemistry Project: "Albuterol Toxicity in Zebrafish and Its Protection with Vitamin E" Abstract: "The goal of this project was to determine the toxic effects of albuterol in an in vivo model system of zebrafish. 24 hpf and 48 hpf zebrafish embryos were exposed to different concentration of Albuterol and were inspected for mortality and heart rate. Significant morphological defect and mortality was observed at concentration of 1000 μM of albuterol treatment. Acridine orange staining of the treated embryos exhibited apoptosis. Concentration dependent increases in superoxide anion (O2-) and nitric oxide (NO) production was also observed as a result of the treatment. Therefore, albuterol treatment suggested the production of reactive oxygen species (ROS) leading to oxidative stress. It was studied if the antioxidant α- tocopherol (vitamin E) has the potential to attenuate the albuterol induced- toxic effects. Zebrafish embryos were treated with a combined mixture of vitamin E and Albuterol. It was found that α- tocopherol has the potential to protect against albuterol-induced effects on the heart rate, apoptosis and reactive oxygen species in zebrafish embryos. Therfore, we suggest that Albuterol in combination with Vitamin E might reduce the toxic effect of Albuterol." Daniel White, Somerset-Berkley High School 3rd place Grand Award - Life Sciences - Medicine and Health Sciences Project: "Utilizing Minkowski Dimension to Efficiently Grade and Diagnose Cancer" Abstract: "The objective of this experiment was to create and test a java-based program capable of more efficiently and accurately assessing the grade of cells thereby automating the process of diagnosing cancer. The cells analyzed were human basal skin cells and the cancer: carcinoma, though the intent is to extend this to a multitude of cell types. The program was designed to work alongside the Philip’s slide scanner with the ability to digitize one slide every 50 seconds. The program utilized a series of image filters to isolate the cells from each of 10 slides. A 10,000- cell sample was randomly selected in which random block design was used to organize the cells into 10-cell partitions eliminating confounding with possible locational nonconformities. Box-counting was used to determine the Minkowski-Bouligand dimension of each cell. Statistical significance was achieved through assembly of a 99% confidence interval in which one may be 99% assured that the population mean of grade 1 human basal skin cells will fall within the range of Minkowski-Bouligand dimensions defined by (1.254,1.345), grade 2 by (1.127,1.254) and (1.345,1.672), and grade 3 by (1.000,1.127) and (1.672,2.000). On average the program grades 10,000 cells in 1.775 seconds, an approximate time-efficiency of 48,670 times that of the average pathologist." Rahi Punjabi, Advanced Mathematics and Science Academy 3rd place Grand Award - Life Sciences - Microbiology Project: "Engineering a Novel Protein Therapy for Meningococcal Infection" Abstract: Neisseria meningitidis is a major cause of meningitis and sepsis worldwide with a 25% mortality rate. Since the bacterium binds to factor H (fH), a protein intended to shield human tissue from the complement system, via factor H binding protein (fHbp), it can elude complement-mediated killing. In this study, recombinant fusion proteins were constructed to bind to fHbp on N. meningitidis and thereby enable the complement system to target the bacterium. These proteins comprised of murine IgG fused to rhesus fH mutants that hypothetically induced a higher affinity for fHbp than human fH. Using an enzyme-linked immunosorbent assay, the binding of the fusion proteins to fHbp was compared to the binding of human fH to fHbp. The rhesus fH G401R mutant fusion protein was able to bind to fHbp at a significantly higher level than the human fH wild type fusion protein. This finding was confirmed using flow cytometry to assess the binding of the rhesus fH G401R fusion protein to representative strains from the A, B, C, W, and Y serogroups of N. meningitidis. Furthermore, there was a significant increase from 41.6% to 81.6% (p<0.01) in the median fluorescence of complement proteins deposited on N. meningitidis across all five serogroups with the addition of the rhesus fH G401R fusion protein. In conclusion, fusion proteins demonstrate considerable potential as a therapy for meningococcal infection since they reverse the complement evasion mechanism of N. meningitidis, boost the body's immune response to infection, and work effectively across a range of meningococcal serogroups. Emory Payne & Zohaib Moonis, Bancroft School 4th place Grand Award - Life Sciences - Cellular & Molecular Biology Project: "Effect of Ethanol on Beta Cell Development in Zebrafish" Abstract: "Excess alcohol during the first trimester of pregnancy has been associated with morphological abnormalities, and may result in pancreatic damage. This study investigated the relationship between Fetal Alcohol Syndrome and Type 1 Diabetes using a zebrafish model. Embryos were exposed to increasing concentrations of ethanol during pancreatic development. Six experimental groups were created exposing embryos to 0.10%, 0.25%, 0.50%, 0.75%, 1.0%, and 1.5% EtOH. The control group exposed embryos to 0% EtOH. The embryos were graded (scale 0-4) based on the level of beta cell degradation in the pancreas; 0 being no cell degradation and 4 indicating complete degradation. Beta cell degradation increased directly proportional to increasing EtOH concentrations, as seen in a decrease in grade 0 and increase in other grades. Zero percent EtOH resulted in 82.35% grade 0 fish while 1.5% EtOH resulted in 0% grade 0 fish. A graded response was observed with increasing EtOH concentrations, resulting in 82.14%, 70.97%, 52.08%, 63.64%, and 8.51% grade 0 fish for other concentrations. As the percentage of grade 0 fish decreased, percentage of fish classified in grades 1- 4 increased. For example, 0.1% EtOH exposure resulted in 82.14% grade 0 fish, 3.57% grade 1, 0% grade 2, 0% grade 3, and 14.29% grade 4 compared to 8.51% grade 0, 36.17% grade 1, 29.79% grade 2, 14.89% grade 3, and 25.53% grade 4 with exposure to 1% EtOH. This study suggests that alcohol consumed during the first trimester of pregnancy, may result in a higher risk for Type 1 Diabetes." Nafisa Wara, Boston Latin School 4th place Grand Award - Life Sciences - Cellular & Molecular Biology Project: "Purification of Mycobacterium Tuberculosis Antigen and Antibody" Abstract: "The goal for the project has three main steps: purify an antigen found to be associated with mycobacterium tuberculosis (mtb) by affinity chromatography, use the purified antigen for antibody purification from the serum of a rabbit raised against this antigen, and then test the specificity of the antigen-antibody interaction with an assay. The gene that codes for the antigen of interest (HuAg6) was cloned into an expression vector for the purpose of my experiment, so the HuAg6 clone was first confirmed by double digestion with restriction enzymes. Then, a western blot was conducted to confirm the presence of the polyhistidine tag on the outside of the antigen when expressed by E. coli host cells. HuAg6 was over-expressed in BL-21 PlysS E. coli and purified by immobilized metal affinity chromatography (IMAC), and an SDS-PAGE was run to confirm its purity. The purified antigen was then coupled to a sepharose resin and used to purify its specific antibody from the serum of a rabbit raised against this antigen. Finally, an immunological assay (ELISA) was used to see how low of a concentration of the antigen is possible in a sample for there still to be detection by the antibody. In the end, the purified antibody specific to HuAg6 is found to still detect the antigen at a 40,000x dilution, which confirms its specificity. This antibody could be used in the development of a less invasive diagnostic for patients infected with mtb, one of the main causative agents of tuberculosis." Erica Budina, Medford High School 4th place Grand Award - Physical Sciences - Engineering: Materials and Bioengineering Project: "Effect of Crosslinking on Mechanical Properties of ECM-Fibrin Scaffold" Abstract: "Heart disease is currently the leading cause of death worldwide. Solubilized extracellular matrix (ECM) is a promising scaffold material that can be utilized to repair damaged areas of the heart. The goal of this project is to create a crosslinked cardiac ECM-fibrin hydrogel scaffold that mimics, as closely as possible, the properties of healthy native myocardial tissue. The approach was to chemically crosslink ECM using the crosslinkers ribose, genipin, and transglutaminase and add fibrin to further improve its mechanical properties. This study was conducted in three phases that sought to crosslink solubilized ECM, ECM and fibrin hybrid scaffolds, and study cell viability within the scaffolds. The degree of crosslinking was evaluated by microscopy for evidence of fiber formation, visible light spectroscopy, and free amino group assays. A uniaxial stretching test was also performed to study the stiffness of ECM-fibrin formulations. The presence of shifts and peaks in the absorbance spectra and the observance of fiber formation via imaging confirmed crosslinking in genipin, ribose, and transglutaminase formulations. The Young’s modulus was increased from 2.6kPa in fibrin-only formulations to 10kPa in ECM-fibrin formulations crosslinked by transglutaminase. When cells were seeded, 57% survived in control samples. Scaffolds crosslinked with 500:1 ECM to TG, 100mM ribose, and 200mM ribose maintained similar cell viability. This study has created a novel scaffold material by crosslinking ECM using various crosslinkers and by adding fibrin. Furthermore, it has also established a method that enables the manipulation of the physical and mechanical properties of ECM hydrogels without significantly affecting cell viability."