Eelgrass (Zostera marina) is a vitally important organism to the Puget Sound that exists along the shallow, muddy seabed. Many different living creatures of all trophic levels rely on the eelgrass beds to survive and absorbed contaminants could be a real threat. Toxics metals in the eelgrass, sediment and water column could have critical implications to the ecosystem. The heavy metal concentrations in the eelgrass, sediment and the water column of two locations in the Possession Sound (Mukilteo and Buoy) were analyzed in order to determine the specific levels of metals and contaminants that travel up through the trophic pyramid. The study endeavors to determine the metal absorbance ability of eelgrass and check if factors including temperature of the water column, seasonality or time of day influenced heavy metal uptake by eelgrass. A sediment grab was utilized to collect the sediment and eelgrass samples. Sample analyses were conducted at the Everett Environmental Laboratory and in partnership with a University of Washington trace metals laboratory to find the concentration of the metals arsenic, copper, lead, zinc and cadmium in the surrounding water column and the eelgrass. It was predicted that greater concentrations of copper and zinc would be found near Buoy because they are legacy chemicals and due to anthropogenic influences near the Everett Port of Everett. It was also hypothesized that a higher concentration of metals would be found in the eelgrass during the spring and summer as more direct sunlight leads to greater primary productivity and uptake of chemical constituents. Determining these types of correlations and catalysts is significant to the entire Possession Sound ecosystem as it would lead to possible strategies on how to prevent further contamination of eelgrass. Furthering our understanding of the connected network of interactions here can only benefit the future inquiries.

Probiotics are an area of significant clinical research, as they have been shown to aid digestion, fight infections, and even mitigate irritable bowel syndrome; however, probiotic colonies must grow and thrive in the extreme pH environment present in the human digestive tract. Previous experiments focused on the production of the surface protein Elongation Factor Thermally Unstable (EF-Tu), which attaches to glycoproteins lining the intestines and has been used as a biomarker for probiotic health. The growth of several strains of probiotics (Lactobacillus bulgaricus, Lactobacillus acidophilus, and Bifidobacterium longum) in both lysogeny broth (LB) and De Man, Rogosa, and Sharpe broth (MRS) was evaluated. The fractions of secreted, cytosolic, and membrane proteins were quantified using a Bradford assay. Finally, the relative amount of EF-Tu was determined by tryptic digestion and liquid chromatography-mass spectrometry (LC-MS). LB was found to be a more effective growth medium for the range of bacteria tested, with 24 hours at 37°C the ideal incubation period. To isolate the secreted proteins, the supernatant was drawn off the broth culture after centrifugation. Freeze-thaw lysis was used to extract cytosolic proteins, with three cycles determined as optimal for protein recover. Sodium deoxycholate was used to separate proteins from the membrane. All three protein samples were separated and run through a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the EF-Tu bands (roughly 43 kDa) were excised and sent for analysis by LC-MS. Using the information obtained from this research study, pharmaceutical companies can create more suitably tailored probiotic products, and make them more accessible and understandable to the general population. 

Oil-spills are a widespread hazardous environmental issue, often caused by mistakes when drilling or transporting crude oil. The effects are devastating to the ecosystem. There are very few ways to quickly clean up an oil spill efficiently, at low cost and without additional environmental damage, as often they are cleaned by using expensive chemical dispersants. We conducted research on an environmentally friendly and inexpensive solution to this problem by using pearl oyster mushroom mycelium. The pearl oyster mushroom has a lignin decomposing basidiomycotina that produces a set of extracellular ligninolytic enzymes that have bioremediation properties. Our hypothesis is that, oyster mushrooms can be used to clean oil spills. We cultivated the mushroom mycelium in straw on trays with different amounts of oil on them. We grew this in a greenhouse with controlled temperature and allowed 4-6 weeks for the mycelium to eat the oil. We are currently observing the results. After 6 weeks, we will filter out the remaining hydrocarbons with a solvent and determine if the mycelium reduced the amount of hydrocarbons or consumed them entirely. Our anticipated results are that the mushroom will clean up the oil in six weeks. For future work, we plan on conducting this same experiment but in a salt water environment to explore applications for mycoremediation in a simulated ocean environment.

Familial Platelet Disorder with Predisposition for Acute Myeloid Leukemia (FPD/AML) is an autosomal dominant human disorder caused by germline, heterozygous mutations in RUNX1. RUNX1 is a master regulator of hematopoiesis and has specific roles in megakaryocyte maturation and the production of platelets. Monoalellic mutations in RUNX1 result in haploinsufficiency of RUNX1 protein, leading to thrombocytopenia and impaired platelet function before leukemic transformation later in life. RUNX1 is controlled through direct positive auto-activation and its half-life is tightly regulated through ubiquitin-mediated proteasomal degradation. Tightly regulated auto-regulatory circuits are known to have capacity as centers for epigenetic cellular memory. Cellular memory effect offers the opportunity to transiently manipulate steady state product levels and observe maintenance that continues the newly attained levels.The goal of this work is to determine if the positive feedback loop controlling RUNX1 expression has capacity for epigenetic cellular memory that is responsive to transient RUNX1 overexpression. We hypothesize that increasing the presence of endogenous RUNX1 in RUNX1-deficient cells will correct megakaryocyte maturation and platelet formation. To initially determine if an increase in the half-life of RUNX1 will correct megakaryopoiesis, a series of ubiquitylnation and proteasome inhibitors were administered to HEK 293t cells for 6hr, 12hr, and 24hr periods to determine their effect of RUNX1 production and stability, showing increases in RUNX1 expression at RNA and protein levels. iPSC's derived from FPD/AML patients will be ultimately differentiated and analyzed via flow cytometry to observe the inhibitors' effects on platelet production. Anticipated results from this work hope to confirm a dose-dependant response in the epigenetic programming of RUNX1 that could illuminate novel therapies for individuals with FPD/AML.

Conventional farming techniques involve large quantities of chemical fertilizers, which often leach into bodies of water causing eutrophication. The influx of excess nutrients from fertilizer results in rapid increase of aquatic algal populations followed by dissolved oxygen depletion. This process creates regions of low oxygen that negatively impact the water quality of many major lakes and coastal regions. To address this problem, the sensitivity of plant growth-promoting bacteria (PGPB) to excess fertilizer was investigated. We hypothesize that the addition of high amounts of fertilizer will result in smaller, less productive plants and diminished rhizosphere colonization of the PGPB, Bacillus subtilis. A modified Kirby-Bauer disk diffusion test was performed as an initial assessment of the effect of varied fertilizer concentrations on B. subtilis. Roots of romaine lettuce seedlings were then inoculated with B. subtilis and grown in soil treated with the same range of fertilizer concentrations. The effects of each treatment on plant growth were determined using total leaf area, quantification of the rhizosphere colonization by B. subtilis, and carbon assimilation measured with a LI-6800 Portable Photosynthesis System. These results are an important step towards establishing guidelines for appropriate application of agricultural fertilizer in order to mitigate the frequency and severity of eutrophication events in aquatic systems.

In the US more than 100 million people are living with diabetes or pre-diabetes. The economic burden caused by these conditions, including medical costs, is approximately $322 billion annually as of 2013. Conventionally, transgenic Escherichia coli has been the primary source of commercial insulin production, a process that requires extensive purification to ensure shelf stability and complete removal of contaminants. This study seeks to establish an alternative mode of insulin production using polyethelyne glycol (PEG) to transform the oyster mushroom, Pleurotus ostreatus, with the human insulin gene. P. ostreatus is a valuable target for genetic transformation due to its lack of endotoxins and fully sequenced genome. P. ostreatus was transformed using PEG with a plasmid containing the human insulin gene and a carboxin resistance gene. Transformed cells were selected using carboxin, extracted, and regenerated on plates composed of yeast extract, malt extract, and glucose (YMG). Integration of the human insulin gene in to the mushroom genome was confirmed through PCR analysis of the transformants. Successful PEG transformation of P.ostreatus offers a new avenue for insulin production, potentially diversifying the market and treatment options for diabetics.