MutY is a bacterial DNA repair glycosylase that cleaves a mismatched adenine across the oxidatively damaged guanine base, 8-oxo-7,8-dihydroguanine (OG), via base excision repair (BER). Unrepaired DNA damage may cause disease-inducing mutations, including changes in transcription regulation or a resultant translated protein. When germ-line mutations occur in the gene of the human homolog, MUTYH, it can lead to a condition called MUTYH associated polyposis (MAP), a colorectal cancer (CRC) subtype. Some variants associated with MAP exist in the interdomain connector (IDC) that joins the N-terminal domain responsible for catalysis to the C-terminal domain necessary for OG recognition. The Q324H variant is located in the IDC in a critical binding domain of several downstream repair proteins. Initial studies with this variant demonstrated reduced OG:A repair in vivo yet maintains wild type (WT) activity in vitro. In this study, we employ the mouse Mutyh homolog of Q324H, Q306H, to better characterize Q324H activity due to difficulties in assessing human MUTYH activity in vitro. Although these studies with the mouse homolog variant may also show WT OG:A repair in vitro, due to the location of this variant in the IDC, it’s CRC association may be caused by perturbations in downstream repair.

The world loses some food crops to pathogenic infections, and investigating how pathogens infect plants would help reduce how much crop is lost. In turn, this leads to the development of new pesticides and pathogen resistant plants. Plants have two main defense systems, which are Pattern Triggered Immunity and Effector Triggered Immunity. The first system includes the plant’s receptors on the outside that trigger the immune response after detecting a pathogen effector. These extracellular receptors act like plants first layer of defense against invading pathogen. Pathogens have evolved sophisticated means to compromise this primary layer of defense by delivering effector protein inside the plant cells. Plant employ a second layer of defense through intracellular Nuclear Binding - Leucine Rich Repeat (NLR) receptors that direct the Hypersensitive Response after detecting an effector. In nature, these effectors are injected into a plant host cell via the Type Three Secretion System. The secretion system complex protrudes out of the bacterial cells and is comprised of a transmembrane base and a long, narrow needle to directly release effectors into a host. Here we explore the localization of these effectors using confocal microscopy. We engineered effector proteins fused to a fluorescent protein by molecular cloning, resulting in a chimeric protein that was delivered into tobacco leaves through Agrobacterium mediated transformation.  Our data reveals that the pathogen effectors localize to the plasma membrane, chloroplast, and mesophyll cells of the tobacco plant leaves.

The severe thunderstorm outbreak of 15 May 2018 devastated large portions of the Northeast with huge swaths of damaging wind up to 110 mph and a dozen tornadoes up to EF2 in strength. Five people were killed and 1.4 billion dollars in damage were caused by the event, making it among the costliest and deadliest severe thunderstorm outbreaks in the history of the Northeast. However, the storm prediction center (SPC) failed to forecast the severity of the event in the days before it occurred, potentially leaving many residents of impacted areas uninformed of the dangerous conditions soon to occur. This case study attempts to answer both why this event was relatively severe and why the SPC failed to forecast the event’s severity. This was accomplished by analyzing synoptic conditions, significant mesoscale events, and model data associated with SPC forecasts. The outbreak was also compared to a previous significant Northeastern outbreak that occurred on 31 May 1998. The current analysis concludes that the severe weather event was an unusual combination of an elevated mixed layer and very high wind shear, due to a split stream pattern (in which the jet stream branches into two different components) and a 700mb trough over the West Coast. This is a synoptic setup that matches the one in 1998 very closely. Model and NWS text data suggests that ample evidence of incipient severe weather existed preceding the event, implying that the SPC could have forecast its severity. It was also determined that the event should be classified as a derecho using procedures outlined in Johns and Hirts, 1987. The split stream setup, when taken with its similarity to past significant severe outbreaks over the Northeast, introduces a better understanding of Northeast severe weather that can aid in forecasting of severe thunderstorm potential.

Macrophages play a key role in maintaining adipose tissue homeostasis, yet recently these cells have become extensively associated with adipose tissue inflammation upon the development of obesity, a disease that affects 2.1 billion people in the world today. In response to the chronic lipid overloading of adipose tissue during the development of obesity, adipose tissue macrophages increase in number and change their localization. This newly attained knowledge shifts medical approaches away from targeting the inflammatory traits of adipose tissue macrophages, towards targeting their metabolic programming. Mediations directed at increasing metabolic capacity might be used to reprogram macrophage metabolism, allowing macrophages to better deal with metabolic challenges during obesity, in order to maintain adipose tissue homeostasis. Taking this into account, the aim of this study was to determine which metabolites produced by Bifidobacterium grown on specific milk oligosaccharides will suppress inflammation induced by the inflammatory stimuli, LPS, in macrophage cells. RAW-Blue cells, a macrophage-like, Abelson leukemia virus transformed cell line derived from BALB/c mice, were cultured and exposed to various Bifidobacterium supernatants produced from Bifidobacterium consumption of lactose, bovine milk oligosaccharides, 2'-Fucosyllactose, and 3'-Sialyllactose. These cells were then induced with LPS, activating the transcription factor, NF-kB. Upon activation of NF-kB in these RAW-blue cells, a secreted embryonic alkaline phosphatase was secreted into the cell supernatant. After overnight incubation, a QUANTI-blue macrophage inflammatory assay was executed in order to detect the levels of alkaline phosphatase. This study provides the first evidence that high concentrations of lactose and 2-FL metabolites were found to significantly decrease NF-kB activation. Therefore, it was concluded that metabolites produced from Bifidobacterium consumption of lactose and 2-FL suppress the LPS induced inflammatory response in adipose tissue macrophages.

Grape splitting has affected the grape and wine industry greatly. Thousands of dollars are spent yearly to cover the cost of grapes that split because they cannot be sold to the consumers or wine companies. Past research only indicates that cracking could be a cause of unevenness of skin surface or the irregularities of pressures within the grape. To inhibit cracking, grapes were spray-painted with white dots randomly on the surface and were then placed in water with randomly spray-painted white dots and analyzed over time. The analysis process consisted of running the images taken of the grapes through MATLAB to produce eigenvalues. The eigenvalues track the percent movement of a centroid of three white dots over time. The eigenvalues were then plotted using SAS and patterns were drawn from the graphs. It was found that eigenvectors in the cracking area were growing in a nonuniform way the moment before the crack. The eigenvectors would become larger in both directions indicating an increasing strain adjacent to where the crack would eventually form. Eigenvectors that were not directly beside the crack but in the cracking area showed a decreasing strain perpendicular to the crack. However, within the same eigenvector there would be an increasing strain perpendicular to the crack. This is caused by the crack pushing the centroids of the dots closer to other dots on the same side of the crack which could be a result of either differences of pressure in that area or an unevenness of skin thickness. 

Intra-cell heterogeneity in breast tumors may confer resistance to conventional cancer treatments. Certain types of cells may be resistant to a certain treatment, thus allowing cancer to persist. We hypothesized that the heterogeneity of  cancer cells can be evidenced in multiple ways, including the identification of discrepancies in their metabolic processes. To this end, we sought to select and characterize a subpopulation of cells that only utilize mitochondrial metabolism to obtain energy from fuels. Adenocarcinoma breast cancer cells (MDA-MB-231) were grown in a galactose or glucose media. The cells in galactose were selected for the ability to use mitochondrial metabolism, while the cells in glucose served as the control. Cell proliferation, phosphorylating capacity, and markers for differentiation, pluripotency, and metastasis were evaluated in cells grown in galactose versus those in glucose. Cells grown in galactose (cells that rely on mitochondrial metabolism including fatty acid oxidation) were found to proliferate slower, have higher ATP production, and were less pluripotent and less differentiated. Thus, breast cancer cells which were forced to generate ATP from mitochondria displayed unique morphological and metabolic characteristics that seem to include a transition towards stemness.

A human and a mouse both have 3.1 billion base pairs which makes a mouse’s genome ideal for genetic research (Importance of Mouse Genome 2017). This project will include the comparison of two important pieces of biotechnology: Taqman assays and traditional PCR, in order to discover which is more cost efficient and accuracy of the screening methods when looking DNA insertions. Analyzing these screening accurate will help provide MBP with the most effective way to produce transgenic and Knockout mice thus furthering scientific research. The Taqman assay and traditional PCR will screen a large Knock in of GFP, a Knock in SNP (single nucleotide polymorphism), and a knock in of loxP/CRE which is a site specific recombinase consisting of around 30 base pairs (Valenzuela et. al., 2003). A Taqman assay is initially more expensive, but it is hypothesized to perform better than traditional PCR when working with small changes in the number of base pairs (Redig 2014). On the other hand, traditional PCR is better suited to find a large insertion than a Taqman assay (Valenzuela et. al., 2003). To investigate whether a Taqman assay or PCR is more effective in certain screening methods, time measurements and cost calculations must be taken into consideration for each of the methods. The quality of the screening must also be taken into consideration to ensure that quality is not being sacrificed for cost or time efficiency.