Fertilization, both in plants and animals, is at its core, a study of cell to cell communication. With respect to plants, the male gametophyte, the pollen tube, elongates within the female organ called the pistil, transporting in its cytoplasm two sperm cells. The pollen tube is attracted by signals secreted from the synergid cells that are located at the entrance to the female gametophyte that resides in the ovule. Secondary pollen tube visitors to the ovules are unwanted and repelled presumably by signals emitted by the fertilized female. The final communication between the pollen tube and female gametophyte is the induction of pollen tube rupture upon penetration of the synergid cell, an event that leads to the release of the two sperm cells, which go on to fertilize the central cell and egg cell within the female gametophyte, completing a double fertilization process that is unique to plants. My thesis research is centered on elucidating the mechanism behind the synergid cell-induced pollen tube rupture process. Studies in our laboratory have established that the synergid cell-expressed receptor like kinase, called FERONIA, mediates a highly oxidative environment in the female gematophyte that is necessary for the pollen tube rupture process. Using an in vitro pollen tube culture system, my research showed that reactive oxygen species (ROS) induces pollen tube rupture in a Ca2+-dependent manner. My results suggests a careful and truly fascinating, though still hypothetical, design of a two molecule, FERONIA and ROS, two step activation system that uses ROS to prime the pollen tube outside the synergid cell, then expose it to calcium within the synergid cell to ensure that pollen tube rupture happens in the synergid cell, enabling fertilization.

Obesity accounts for $168 billion in annual medical expenses and increases the risk of cardiovascular disease, cancer, and type-2 diabetes, three diseases responsible for over 50% of deaths in the United States. It is well established that the pattern of adiposity is an important factor in the relationship with disease risk and that visceral adiposity, which favors hypertrophy (characterized by enlarged cells) is more dangerous than subcutaneous adiposity, which tends to be hyperplastic (characterized by an increase in cell number). Hypertrophy is associated with inflammation and insulin resistance, and hyperplasia (adipogenesis, i.e., the formation of new adipocytes), is associated with improved insulin sensitivity. Tumor necrosis factor-alpha (TNF-alpha) is a potent pro-inflammatory cytokine that activates a nuclear factor-kappa B (NFKB) intracellular pathway that is an important mediator of obesity-associated insulin resistance and increased risk of type-2 diabetes. Interestingly, obesity has been positively associated with both low vitamin D status and elevated levels of TNF-alpha. Our studies focused on examining the influence of the active vitamin D hormone, 1,25-dihydroxyvitamin D, and TNF-alpha on adipogenesis and inflammation in human primary adipocytes and determining whether the balance of these two factors influences the extent to which adipocytes accumulate lipid or express pro-inflammatory cytokines. We found no effect of 1,25-dihydroxyvitamin D on adipogenesis or pro-adipogenic gene expression despite a clear upregulation of a vitamin D responsive gene, 24-hydroxylase, in response to treatment with 1,25-dihydroxyvitamin D. TNF-alpha clearly inhibited adipogenesis and expression of PPAR-gamma and C/EBP-alpha and enhanced expression of the pro-inflammatory cytokines IL-6 and MCP-1, but not IL-8. There was a trend towards a dose-dependent downregulation of MCP-1 by 1,25-dihydroxyvitamin D in three individuals; however, this effect was not statistically significant. While we found no interaction between TNF-alpha and 1,25-dihydroxyvitamin D on adipogenesis, there is a potential anti-inflammatory action of 1,25-dihydroxyvitamin D in human primary adipocytes. Future studies into this potential are warranted in light of the growing obesity epidemic and the interest in finding nutritionally modifiable treatment or prevention strategies to mitigate the negative consequences of obesity.

Commercial carrot processors produce up to 175,000 tons of carrot waste annually. Carrot Mash (CM) is the term referring to the waste by-product of peeled baby carrot processing. Transportation of carrot processing waste is expensive due to its high- water content (approx. 83-95%). High in bioactive compounds (carotenoids) and dietary fibers, it is expected that its conversion into a value-added by-product is of interest to the carrot processing industry. Hemicellulose-rich plant materials have proven to be a source of oligosaccharides, which are known for their beneficial prebiotic activity. The objectives of this research were to: 1) determine the effect of mechanical treatments on the extraction of water and bioactive compounds and evaluate the functional properties of carrot mash; 2) incorporate dried carrot mash into a beef patty and evaluate changes in pH, color, cooking yield, and texture; 3) apply an enzymatic treatment to carrot mash to promote the conversion of polysaccharides to oligosaccharides for prebiotic benefits. Mechanical separation of liquid and solid fractions by way of expeller pressing was efficient in extracting liquid while simultaneously increasing total solids by nearly 200%, the extraction of carotenoids by 1000%, and polyphenol content by nearly 97%. Mechanical treatments increased the fat binding capacity on average by 183% compared to untreated mash. The addition of unpressed carrot mash or expeller pressed carrot mash increased the cooking yield of a beef patty by 3-13% without significantly changing its textural properties. Enzymatically treating the carrot mash significantly increased the concentration of oligosaccharides up to 2.3%. These results suggest that carrot processing wastes can be physically and enzymatically modified and have an immense potential to be utilized as a functional ingredient in human food rather than being landfilled, composted or used as animal feed.

Matthew R. Borglin This thesis demonstrates efficacy of Octenyl Succinic Anhydride (OSA), as a biofilm sanitizer. Biofilms allow bacteria to adhere to solid surfaces with the use of excreted polymeric compounds. For example, surfaces found in food production or processing facilities such as the interior of a raw milk holding tank, are some of the most susceptible to biofilm contamination. When present, biofilms can cause a variety of negative effects, which include; reduction of product shelf life, corrosion, and outbreaks of foodborne illnesses. The close association of biofilms with the majority of foodborne illness cases led the US Environmental Protection Agency (EPA) to create a new category of sanitizer specifically designed for treatment of mature biofilms. The efficacy of sanitizers in this new regulatory category is determined by the EPA protocols MB-19 and MB-20. The EPA’s protocols outline methods for cultivating, treating, and measuring effects on Pseudomonas aeruginosa biofilms in a continuous flow stir bar bioreactor. Biofilm modification by OSA was verified by the presence of octenyl esters on OSA treated biofilms with single point Raman spectrophotometry. OSA modified biofilm’s antimicrobial properties were first investigated with crystal violet staining in 96-well microtiter plates with inconclusive results. However, effective antimicrobial properties where apparent when using the CDC Biofilm Reactor. OSA treatments consistently returned a 6-log CFU/coupon reduction in biomass compared to controls. Inhibition of planktonic and/or biofilm regrowth was demonstrated using the 96-well plate methodology. This thesis demonstrated the effectiveness of OSA chemical esterification reaction as a biofilm treatment. In doing so, this work suggests a new approach for biofilm remediation by chemically modifying the structural components of biofilm.

Humans experience ubiquitous exposures to estrogenic environmental chemicals from food, personal care products, and other industrial and consumer goods. Bisphenol A (BPA), a well-studied xenoestrogen, is known to alter development of estrogen-sensitive organs including the brain, reproductive tract, and mammary gland. Bisphenol S (BPS), which has a similar chemical structure to BPA, is also used in many consumer products, but its effects on estrogen-sensitive organs in mammals has not been thoroughly examined. In our study, pregnant CD-1 mice were orally exposed to BPS or ethinyl estradiol (EE2, a positive control for estrogenicity) from gestational day 9 through postnatal day (PND) 2, the period when many estrogen-sensitive organs are developing. After weaning, the offspring were administered either oil (vehicle) or an estrogen challenge (1 μg EE2/kg/day) for ten days starting at PND21 (prior to puberty), PND80 (early adulthood), or PND260 (later adulthood). Timing of puberty was evaluated in females by noting the date on which vaginal opening occurred. After the 10 day estrogen challenge, we evaluated the response of endocrine sensitive organs through measurements of organ weight, tissue morphology, and gene expression in both males and females. We observed dose- and sex-specific effects of BPS and EE2 treatment, as well as alterations in the responses of males and females to the estrogen challenge. This study sheds light on the effects of low dose xenoestrogen exposures on estrogen-sensitive organs including the reproductive tract and mammary gland. Furthermore, it improves our understanding of the influence of environmental chemicals on secular trends of earlier age of puberty in girls reported over the past few decades.

Glioblastoma is the most commonly diagnosed central nervous system primary malignancy and it is considered a terminal diagnosis with few treatment options available. Glioblastoma tumors frequently develop treatment resistance due in part to their highly heterogenic nature. The heterogeneity of glioblastoma is partially attributed to the presence of glioma stem-like cells (GSC), which are highly invasive and resistant to chemotherapy and irradiation treatments. Signaling of the receptor tyrosine kinase MET is a known regulator of GSC. Glioblastoma patients have an increasingly poor prognosis that corresponds with increasing MET expression. Both GSC and MET are known to contribute to treatment resistance in glioblastoma and several MET alterations have been observed in glioblastoma. In these studies, we investigated MET alterations that are commonly found in glioblastoma. Using patient-derived xenograft (PDX) lines, the MET alterations were characterized and confirmed to be MET positive, MET amplified, or harbor a PTPRZ1-MET fusion. We also included a MET null glioblastoma PDX line. The PDX lines demonstrated markers for GSC potential with all showing neurosphere formation, the ability to initiate tumor growth in immune-compromised mice, and expression of GSC markers GFAP, Sox2, and nestin. The MET alterations were further examined by examining tyrosine kinase inhibitors' effect on viability and MET signaling. Oncogene addiction through MET amplification was found to have the best response to inhibition. The MET fusion bearing line demonstrated less sensitivity to inhibition than has been shown in other studies, indicating a need for further research into co-mutations that increase sensitivity to MET inhibition. We also investigated the efficacy of novel MET-targeting chimeric antigen receptor T cells (MET.CART cells). The MET.CART cells were able to specifically target and successfully kill MET-expressing glioblastoma cells. Together these results imply the need for more personalized treatment of glioblastoma based on the molecular biology of the tumor.

Sessile invertebrates in the rocky intertidal experience intermittent periods of air exposure due to tidal flux, presenting risks of temperature extremes, hypoxia, nutrient limitation, and most dangerously, desiccation. Microscale variation in severity and frequency of these risks is widely dependent on vertical position within the intertidal zone. Common acorn barnacles (Balanus glandula) have a wide vertical distribution in the intertidal, creating large differences in microhabitat between the highest and lowest individuals in the population. This study set out to explore whether tidal position dependent differences exist in the response to oxygen and desiccation stress in B. glandula. We hypothesized that B. glandula from relatively high tidal heights, which are exposed to the air for a greater duration, will be better suited to tolerate anoxic and desiccation stress than conspecifics from lower tidal heights. To explore this, we compared responses of B. glandula collected from high and low intertidal positions to A) anoxia (0 mg O2/L) and hypoxia (≤ 2 mg O2/L) on survival, behavior (closed opercular plates, cirral beating, pneumostome formation), enzyme activity (lactate dehydrogenase (LDH), superoxide dismutase (SOD)), and tissue-lactate accumulation, in addition to B) the effects of humid (98% RH) and dry (32% RH) air emersion (at 17˚C) on survival, opercular behavior (open/closed), evaporative water loss (EWL) rates, and tissue-lactate accumulation. Relative to barnacles from the low intertidal, we found that barnacles from the high intertidal survive longer during anoxia and air emersion stress, close their operculum sooner in dry air, lose more water during air exposure at any humidity level, and tend to accumulate less D-lactate. We suspect that high intertidal B. glandula can survive desiccation longer by ejecting stores of mantle cavity fluid, thereby creating a moist lung-like, air-filled internal environment, then remaining largely closed and metabolically inactive when in air to avoid drying out and becoming anoxic. These differences may reflect plasticity or selective pressure in response to environmental stress during development and highlight the potential importance of microscale stress heterogeneity in influencing species climate change tolerance and potential distribution patterns.

<p>The aim of this thesis is to study the role of banking in society and the effect of the</p> <p>interbank market on the performance of the banking system.</p> <p>It starts by reviewing</p> <p>several studies conducted on empirical banking networks and highlighting their salient</p> <p>features in the context of modern network theory. A simulated network resembling the</p> <p>characteristics documented in the empirical studies is then built and its resilience is</p> <p>analyzed with a particular emphasis in documenting the crucial role played by highly</p> <p>interconnected banks.</p> <p>It is our belief that the study of systemic risk and contagion in a banking system</p> <p>is an integral part to the study of the economic role of banks themselves. Thus the</p> <p>current work focuses on the fundamentals of banking and aims at identifying the</p> <p>necessary drivers for a dynamical setup of the interbank market.</p> <p>Through an agent–based model, we address the issues of bank formation, bank runs</p> <p>and the emergence of an interbank market. Starting with heterogeneous individuals,</p> <p>bank formation is viewed as an emergent phenomenon arising to meet the needs for</p> <p>investment opportunities in face of uncertain liquidity preferences. When banks work</p> <p>in isolation (no interbank market), in the long run and through a long experience with</p> <p>bank failures, banking turns into a monopoly or a market with few players.</p> <p>By equipping banks with their own learning tools and allowing an interbank market</p> <p>to develop, fewer bank failures and a less concentrated banking system are witnessed.</p> <p>In addition, through a scenario analysis, it is demonstrated that allowing banks to</p> <p>interact does not weaken the banking system in almost all the cases, and improves</p> <p>the performance on multiple occasions.</p> <p>The work is concluded by studying the effects of a banking system on individuals</p> <p>and the economy in what is called social measures. We establish that the effects</p> <p>of banking on social measures such as consumption level, consumption inequality</p> <p>between individuals, long term investment and economic waste, varies significantly</p> <p>based on the structure of the society.</p> / Doctor of Philosophy (PhD)

Microplastics are an emerging environmental contaminant. One of the ways microplastics can get into the environment is by the breakdown of larger plastics. These plastics can come from industrial practices, discarded fabrics, agriculture, and general plastic waste. As these plastics are broken down microplastics leach into the environment. The widespread use of plastics has resulted in the spread of microplastic contaminants all over the world. Microplastics have been reported to be in drinking water, so this paper is looking at the presence of microplastics in local communities of different demographics and socioeconomic statuses. In other studies of different drinking water contamination, minority, and low-income communities had worse quality water. In this paper, methods for sampling, extraction, and analysis with Laser Direct Infrared Spectroscopy were used. Out of Cities A, B, and C, City A, with the largest population, higher percentage of minorities, and worse off socioeconomic status, had the highest particle count. More research is needed in this field to find how socioeconomic status can affect exposure to microplastic contamination.

Skeletal muscle fatigue is the contraction-induced decline in whole muscle force or power, and can be greater in older versus young adults. Fatigue primarily results from increased metabolism elevating phosphate (Pi) and hydrogen (H+), which alters myosin-actin interactions; however, which steps of the myosin-actin cross-bridge cycle are changed and their reversibility are unclear. PURPOSE: This study sought to: 1) Examine the effects of elevated Pi and H+ on molecular and cellular function, and 2) Test the ability of deoxyadenosine triphosphate (dATP), an alternative energy to adenosine triphosphate (ATP), to reverse the contractile changes induced with high Pi and H+. METHODS: Maximal tension (force/cross-sectional area), myofilament mechanics and myosin-actin cross-bridge kinetics were measured in 214 single fibers (104 type 1) from the vastus lateralis of eight (4 men) healthy, sedentary older adults (71±1.3 years) under normal (5 mM Pi, pH 7.0), simulated fatigue (30 mM Pi, pH 6.2) and simulated fatigue with dATP conditions. RESULTS: Tension declined with high Pi and H+ in slow- (type I, 23%) and fast-contracting (type II, 28%) fibers due to fewer strongly bound myosin heads (28-48%) and slower cross-bridge kinetics (longer myosin attachment times (ton) (18-40%) and reduced rates of force production (18-30%)). Type I myofilaments became stiffer with high Pi and H+ (48%), which may have partially mitigated fatigue-induced tension reduction. Elevated Pi and H+ with dATP moderately improved force production similarly in both fiber types (8-11%) compared to high Pi and H+ with ATP. In type I fibers, high Pi and H+ with dATP returned the number of myosin heads strongly bound and ton to normal, while the rate of force production became faster than normal (16%). In type II fibers, high Pi and H+ with dATP did not change the number of myosin heads bound, but cross-bridge kinetics were 16-23% faster than normal. CONCLUSION: These results identified novel fiber-type specific changes in myosin-actin cross-bridge kinetics and myofilament stiffness that help explain fatigue-related force reduction in human single skeletal muscle fibers as well as an alternative energy source that partially to fully reverses contractile changes of elevated Pi and H+ that occur with fatigue.