Student Abstracts: Biology at BNL

Teaching about Nature in Nature Integrating Field Methods into Biology Classrooms. JENIFER BERRELLI (UMass Amherst, Amherst, MA 01003) TIMOTHY GREEN (Brookhaven National Laboratory, Upton, NY 11973) .
Presently, most educational curriculums lack active learning and field methods. To enhance learning in high school biology classrooms, it is possible to introduce professional field methods to educators, with hopes that they may integrate more field trips and outdoor activities into their curriculum. This summer's (2001) field program enabled high school students to learn many scientific techniques, skills and concepts. Throughout the summer, the students learned valuable observation skills as they explored numerous nature centers, beaches, dunes, marshes, woodlands, rivers and ponds. Students learned about glacial deposition as a means of land formation, coastal change and development, how to determine history of a coastal, marshland or woodland area given topographical, sediment and/ or vegetational data, how to determine the susceptibility of an area to burn dependant on fuel load, how to determine the difference between areas of natural formation as opposed to areas disturbed by anthopomophic activities, and how to conduct a vegetation and topographical analysis of a segment of river. They proceeded to choose a specific research topic and use the scientific method to evaluate topic of concern, hypothesize, record data, analyze, and conclude the research followed by the production of research papers, posters and presentations. As parents and teachers observed the students' presentations, it is anticipated that these field methods will be regarded as particularly productive and may influence those educators to include more of this type of teaching methods in their program of study.

Is Ycf9 the Linchpin of Photosystem II? Heterologous Expression for Structural and Functional Studies. HELEN CHUNG (Cornell University, Ithaca, NY 14853) GEOFFREY HIND (Brookhaven National Laboratory, Upton, NY 11973) .
One group of ORF's, ycfs (hypothetical chloroplast reading frames), have prove to be highly conserved between species of plants, and are thought to be genuine genes. ycf9 in particular has caught the attention of scientists. It encodes for a 62 amino acid protein that contains two alpha helices that are highly hydrophobic and is found in the PS II thylakoid membrane fractions. Gene knockout experiments have been done to determine the function and importance of this gene while fractionation experiments have been done to locate the whereabouts of its products. But no experiments have been done to isolate the protein in attempt to study its structure. In this experiment, I isolated ycf9 from the leaves of a spinach plant, expressed it in E. coli, and identified it through SDS PAGE. Circular dichroism spectroscopy and polarized infrared spectroscopy will be performed in hopes to unravel one of the many mysteries of photosynthesis'how PSII is put together and/or held together by ycf9.

Purification of an Adenovirus Proteinase Homolog from the Chlamydia Genome. CHRISTINE EMIGH (University of California, Santa Cruz, Santa Cruz, CA 95064) WALTER F. MANGEL (Brookhaven National Laboratory, Upton, NY 11973) .
Chlamydia are bacterial pathogens whose representatives are widely distributed in nature, and C. trachomatis causes several human diseases of medical significance. This is the most common type of genital tract infection and is one of the most damaging of the venereal diseases (Science 1999). Purification and characterization of such a common and diverse type of virulence factor could lead to discoveries that are applicable to all types of virulence factors. It is essential for a good protocol to be designed for the mass purification of this protein for these studies to be carried out. Diffraction data collected from a crystal of the Chlamydia protein could yield important data for the design of a specifically-targeted drug. Although Chlamydia is treatable by antibiotics, this is an expensive and potentially risky treatment. A specifically targeted molecule could end up much less costly to produce and without the potential risks that come with antibiotics.

CCI Enrichment Program . SEBASTIAN GALAZKA (Housatonic Community College, Bridgeport, CT 06604) GLENN WILLIAMS (Brookhaven National Laboratory, Upton, NY 11973) .
The Brookhaven National Laboratory internship program is designed to attract highly motivated students from various colleges to come and experience what it is like to be a researcher in a scientific field. The main goal of the program is to produce a core of scientists, in various fields, that will lead American research into the twenty-first century and continue its high quality and standards as the world leader in scientific research. However, instead of giving the students a closed look into only one specific discipline, the laboratory has created a program designed to broaden the knowledge of the students into the various aspects of all science. This is the Community College Institute (CCI) enrichment program. It is a program that gathers all of the CCI students once a week and expose then to different aspects of science every time they meet. It also serves as an open forum giving the students the opportunity to discuss, on a more personal level, the experiences and problems that they were having. It also allowed the students to see the many aspects of the laboratory through a number of tours that were given by the students themselves.

Viscosity Reduction of Heavy Crude's Aided by Microorganisms. ERICA GOODRICH (Community College of Rhode Island, Warwick, RI 02886) MOW LIN (Brookhaven National Laboratory, Upton, NY 11973) .
Eight sandstone cores were prepared and filled with samples of a heavy crude oil. The samples were treated with saltwater-mimic and two different strains of Bacillus species of microorganisms at 60oC. After treatment the following parameters were measured. These were: 1) Quantity of oil extracted, 2) viscosity, and 3) hydrocarbon distribution. The treated samples showed a higher quantity of recovered oil, a reduction in viscosity, and an enhancement of lighter hydrocarbon fractions. A direct comparison of the gas chromatographic data showed that the intensity of the hydrocarbon peaks were higher in the bacteria treated samples as compared to those from the saltwater-mimic sample.

Cholesterol Management: A workshop for reducing a major risk factor for heart disease.. MIRIAM HERNANDEZ (LaGuardia Community College, Long Island City, NY 11101) MARY WOOD (Brookhaven National Laboratory, Upton, NY 11973) .
Heart disease is the major killer of women and men in the United States. And cholesterol is one of the major factors for heart disease. Cholesterol is a fat-like substance that when in high levels, causes the blood vessels to narrow and eventually blocks the flow of blood leading to a heart attack or heart disease. For this reason, a three-month cholesterol workshop (CW) was run for BNL employees in order to control and maintain their cholesterol levels. The CW provided information about cholesterol importance in our body, cholesterol as a risk factor for heart disease, cholesterol types and measurements, cholesterol food sources and cholesterol-lowering treatments. All participants had their lipoprotein profile (cholesterol measurements) at the beginning and end of the CW. Then the two cholesterol measurements of each participant were compared to identify changes in their cholesterol levels. Moreover, each participant filled out a final evaluation to determine their lifestyle changes done after the CW. Looking at the data of the participants, it was found that many participants become more informed about cholesterol issues and changed their lifestyles. Few participants even reached desirable cholesterol levels. These findings suggest that cholesterol levels can be controlled by correct information and lifestyle changes. In this way, heart disease can be prevented.

Herbicide Selection of Transgenic Plants. SOPHIA LIN (State University of New York at Stony Brook, Stony Brook, NY 11794) DR. JOHN SHANKLIN (Brookhaven National Laboratory, Upton, NY 11973) .
Although herbicides are commonly used to control weeds, they also have excellent potential to be used for the selection of transgenic plants. The PAT gene confers resistance to the herbicide glufosinate ammonium (GLA). Adding the PAT gene to a vector with other genes of interest would allow selection by means of a simple herbicide treatment. Seeds without foreign DNA (the PAT gene) would die; survivors would be successful transgenic recombinants. This project used a commercial glufosinate herbicide for selection of transgenic Arabidopsis thaliana. The gene of interest was the castor hydroxylase gene that codes for the hydroxylase that converts oleic acid into ricinoleic acid (a novel fatty acid with commercial value). This type of selection yielded many transgenic plants that were both resistant to GLA herbicide and produced novel hydroxy fatty acids.

Is Ycf9 the Linchpin of Photosystem II? Heterologous Expression for Structural and Functional Studies. EMILY SHESTON (Wilkes University, Wilkes-Barre, PA 18766) DR. GEOFFREY HIND (Brookhaven National Laboratory, Upton, NY 11973) .
The ycf9 gene (orf 62) is found in chloroplast genomes of all plants as well as in cyanobacteria. When Ycf9 is phosphorylated by an intrinsic protein kinase (Race & Hind, 1996), it detaches from photosystem II, allowing migration of LHC-II away from the reaction center core diminishing reaction center energy capture. Work done by Ruf et al. (2000) showed that Ycf9 mutants also lacked a mobile chlorophyll a/b protein, CP26, presumably because CP26 could not be assembled into the photosystem. We hypothesize that Ycf9 is a linchpin holding CP26 and LHC-II to the reaction center core of photosystem II. Purification of Ycf9 was attempted using two methods: gel analysis and vector-induced protein expression. Fractions of photosystem II-enriched membrane complexes were run on 16.5% tricine/bis-tris propane acrylamide gels suitable for separating low molecular weight proteins. We designed tricine/bis-tris propane buffer to avoid the complications tris/glycine buffers pose in subsequent protein sequencing. We identified a protein of approximately 6.3 kD in the reaction center cores and photosystem II-enriched subfractions that we suspect to be Ycf9. Chloroplast DNA was prepared from Spinacia oleracea as described by Triboush et al. (1998). PCR was used to amplify the known ycf9 nucleotide sequence and add restriction sites for ligation into the vectors pFLAG-ATS and pFLAG-CTC for expression in E. coli. Low levels of protein were expressed in the periplasm of pFLAG-ATS transformants. Further work will attempt to improve yields of expressed protein and explore other vector systems.

ULTRAVIOLET A WAVEBANDS INDUCE DIFFERENT DNA DAMAGES IN XIPHOPHORUS MACULATUS SKIN CELLS WITH AND WITHOUT MELANIN. TSUHAO YEUNG (UNIVERSITY OF ROCHESTER, ROCHESTER, NY 11953) R.B. SETLOW (Brookhaven National Laboratory, Upton, NY 11973) .
It is known that UVB wavebands (280 - 320 nm) have the capability to induce DNA damages, such as cyclobutane pyrimidine dimers, while the effects of UVA (320 - 400 nm) remain mysterious. If unrepaired, these damages negatively affect the cell's ability to replicate and its homeostatic functions, resulting in cell death and cancerous growth. In our studies, we use UV-endonuclease from Micrococcus luteus and alkaline gel-electrophoresis, to identify occurrences of damage to DNA isolated from black and white skin of Xiphophorus maculatus 163B after exposure to UVA wavelengths over 320 nm. Our preliminary results show that damages due to UVA exposure are readily occurring, especially in skin with melanin, and warrant further investigations.