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ENZYMATIC DEWATERING OF DIGESTED SLUDGE Diane Mar Program: NSF EPSCoR Mentor: Charles Coronella Department: Chemical Engineering University of Nevada, Reno Traditional dewatering processes for sewer sludge are expensive and inadequate for the development of efficient energy production from dried sludge biosolids. While some of the moisture can be removed by mechanical processes, a majority of it is bound intracellularly and by intermolecular forces. Even after dewatering, the moisture content of digested sludge is high at around 85%. Enzymes may be able to assist in dewatering through lysing of cells and releasing some of this bound water, and the ability of proteases, cellulases, and lipases to enhance mechanical dewatering was examined. The enzymes were added to sludge solutions of 5 wt% solids both individually and in combination with each other and were tested at pHs of 5 and 8. The effect of salt concentration in conjunction with enzymes was also studied. Samples were then centrifuged and moisture content measurements through weight loss from oven drying were taken to determine the efficiency of these enzymes. However, enzymes that were tested had little to no effect on the dewatering of sludge with a less than 1% difference on average between the controls and the enzyme treated samples. While enzymes could be an economically beneficial and effective treatment of sludge, further research on the substrates and the enzymes that act upon them would be necessary. THE DETERMINATION OF HOW DOCOSAHEXAENOIC ACID AND LINOLEIC ACID ALTER EICOSANOID PRODUCTION IN A MODEL OF HUMAN LUNG CANCER Clarissa Martins Program: NIH INBRE Mentors: Keith Kikawa, Ronald Pardini Department: Biochemistry and Molecular Biology University of Nevada, Reno It is clear that the omega-3 polyunsaturated fatty acid (PUFA) docosahexaenoic acid (DHA) exhibits in vitro growth inhibitory effects on the A549 human lung adenocarcinoma cell line. Conversely, the omega-6 PUFA linoleic acid (LA) upregulates cell growth rates in the same model of lung cancer cell lines. PUFAs sere as precursors for eicosanoids and this has long been thought to play a role in their effects on various cancers. The omega-3 PUFAs tend to be precursors for anti-inflammatory eicosanoids, while omega-6 PUFAs tend to be converted into pro-inflammatory eicosanoids like prostaglandin E2 (PGE2). In the present study, the effect of DHA and LA supplementation on eicosanoid levels in the human A549 lung adenocarcinoma and BT-474 breast ductal carcinoma cell lines has been examined. LA treatments upregulated PGE2 levels significantly in both cell lines, and these effects correlated with alterations in cyclooxygenase 2 (COX-2) activity levels. Future studies will examine whether there are modifications in the levels of transforming growth factor α (TGF-α) dependent on PGE2, to determine if PGE2 is upregulating pro-proliferative signals. In addition to the findings with PGE2, it was determined that DHA supplementation upregulated thromboxane B2 (TXB2) levels. TXB2 is a stable metabolite of thromboxane A2 (TXA2), which inhibits cyclooxygenase activity, which is typically associated with aggressive cancers. Taken together, these data indicate LA is upregulating cancer cell growth in part by increasing COX 15
activity and PGE2 levels, while DHA may be having inhibitory effects on cancer cell growth by decreasing activated COX. EXPRESSION OF POLY-ADENYLATED NUCLEAR RNA ELEMENT IN KAPOSI’S SARCOMA ASSOCIATED HERPES VIRUS USING LENTIVIRUS Nana Matsumoto Program: NIH INBRE Mentors: Cyprian Rossetto, Gregory Pari Department: Biochemistry and Molecular Biology University of Nevada, Reno Kaposi’s Sarcoma Herpes Virus (KSHV) is a large gammaherpesvirus, which is known to be a causative agent for Kaposi’s sarcoma (AIDS associated cancer), primary effusion lymphoma (PEL), and multicentric Castlemen’s disease (MCL). Therefore, deciphering the virus’ mechanism of replication and its interactions with the host cell is of great interest. One notable lytic transcript produced by KSHV of our interest is PAN (poly-adenlyated nuclear transcript). PAN (1077bp in length) is an exclusively nuclear, unspliced, non-coding RNA that becomes the most abundant transcript within the host cell’s nucleus during lytic infection (~70-80% of total transcript). To date, no known function of PAN is observed. The two objectives of this study were to: first, successfully clone PAN into a vector compatible with lentiviral expression system, and second, obtain viral titers expressing PAN-RNA to transduce target cells in the future. This system allows observing the cellular effects from PAN without any interruptions via other viral components of KSHV, and ensures transfection of a wide spectrum of cells – thus creating a better model that mimics natural infection. To meet the objectives, the PAN locus was PCR amplified and cloned into an expression vector containing the lentivirus genome and antibiotic resistance necessary for selection. The vector, along with other components of the lentivirus genome were cotransfected into human embryonic kidney (HEK) 293FT cells where lentiviral titers were confirmed. After conducting antibiotic selection with HEK 293L cells followed by RT-PCR, we confirmed that PAN-RNA was indeed being produced in the target cells. This suggests that we will now be able to transduce various cell lines with PAN. Future studies will focus on the precise role of PAN and its potential roles in inhibiting the host’s innate immunity. DETERMINATION OF A SORPTION MECHANISM FOR 17B-TRENBOLONE Samantha McBride Program: NSF EPSCoR Mentors: Edward Kolodziej Department: Civil and Environmental Engineering University of Nevada, Reno Trenbolone acetate is frequently injected into cattle as a growth promoter in the beef industry. Once injected, it metabolizes to its most active form, the 17β isomer. After trenbolone (TB) makes its way through the body, it is excreted as a combination of 17β, 17α, and trendione. These metabolites often enter natural waters via rainfall run-off, and have the potential to enter our water supply. Both case studies and extensive research have shown that TB acts as an endocrine disruptor in fish, causing trans-genderization. As a result, the potential impact on human health if these steroids were to enter our water supply is of concern. My project therefore focused on developing a deeper understanding of how 17β-TB moves throughout the environment. Previous research has 16
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Page 19 and 20: HABITAT PREFERENCES OF NORTHERN FLY
Page 21 and 22: global climate change, which may af
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