Effects of Melatonin on Heartbeat and Possible Identification of a Melatonin Receptor in Drosophila Melanogaster

Chapter 1 of this manuscript is a literature review that serves as an introduction to the entire dissertation. Chapter 2 examines the effects of the melatonin injection on heart rate and rhythmicity in Drosophila melanogaster Canton-S (wild-type) pupae and pupae bearing a variety of heart mutations. Chapter 3 investigates further the possible mechanisms of melatonin’s ability to increase heart rhythmicity without significantly affecting heart rate. A melatonin antagonist, luzindole; a high-affinity melatonin agonist, 2-iodomelatonin and RNAi techniques are used to identify a possible melatonin receptor in Drosophila melanogaster. An appendix contains a previously published manuscript detailing experiments performed at the beginning of my graduate studies before I began my work with melatonin. The paper is entitled The Relationship of Heart Function to Temperature in Drosophila melanogaster and its Heritability (Jennings et al., 2009)

Domestic Asset Protection Trusts: Ushering in the Klackaba Era

The growth in the U.S. economy has allowed Americans to increase their savings--but how? A novel approach has emerged in seventeen states: domestic asset product trusts (DAPTs). DAPTs are self-settled spindthrift trusts that allow the settlor to retain a beneficial interest in the trust while removing it from the reach of future creditors. Through the lens of the favorable ruling in Klackaba v. Nelson, this Note addresses why DAPTs should be regarded as an effective method of protecting a settlor’s money and argue for more states to follow suit

Antitrust Consideration in Making Investment in the United States

The antitrust laws of the United States have taken on an increasingly significant role with regard to acquisitions and investments generally, and must be taken into consideration by a foreign investor interested in making foreign investments in the United States. Where the requisite contracts exist to establish subject matter jurisdiction, the antitrust laws of the United States will be applied to all proscribed acts regardless of the nationality of the participants. It is clear that most foreign investment in the United States constitutes the requisite minimum contacts required for jurisdiction. There are two primary antitrust areas which should be addressed by those counseling foreign investors interested in investing in the United States. First, attention should be given to antitrust matters relating to the initial acquisition of assets in the United States. Second, foreign investors should receive counsel regarding general prohibitions contained in the United States antitrust laws, particularly where United States law applied more stringent or different requirements than typical antitrust laws in foreign countries. Two statutes are of primary concern in this analysis. The Sherman Act prohibits any contract, combination or conspiracy that creates an unreasonable restraint on trade, and restricts monopolies or attempts to monopolize. These provisions apply to all trades or commerce amoung the several states or with foreign nations. The Clayton Act proscribes price discrimination, ties-ins and exclusive dealings, and, of most significance for this analysis, bars the acquisition of stock or assets where the effect is to substantially lessen competition in a line of commerce

The relationship of temperature to pupal development rates of the western cherry fruit fly, Rhagoletis indifferens Curran

Adult emergence studies conducted with Rhagoletis indifferens Curran reveal that diapause development in this insect is completed most rapidly at approximately 3°C. These results coupled with those reported by Brown (1978) confirm that chilling at 3°C. for ca. 200 days is best when high percent emergence, good synchrony of emergence, and relatively rapid completion of diapause development is required. Studies comparing emergence of field collected insects with that of insects chilled in the laboratory show that in 1978 diapause development in the field was near completion by the end of February. Similar studies may be necessary to pinpoint the end of diapause in the field population in any given year. A standard method was used to determine the thermal developmental threshold for use in degree-day models for adult emergence. This method yielded a value of 10.2°C. A slightly modified version of this standard method yielded a threshold of 11.7°C. A third method was used which entailed weighting data on the basis of the proportional occurrence of experimental incubation temperatures in the field. This method yielded a threshold value of 8.3°C. Comparison of the three thresholds using field emergence and temperature data for a five year period indicated that a model based upon the 8.3°C base threshold performed with less variation from year to year than did models based upon the other two thresholds

Isolation of CD133+ Liver Stem Cells for Clonal Expansion

Liver stem cell, or oval cells, proliferate during chronic liver injury, and are proposed to differentiate into both hepatocytes and cholangiocytes. In addition, liver stem cells are hypothesized to be the precursors for a subset of liver cancer, Hepatocellular carcinoma. One of the primary challenges to stem cell work in any solid organ like the liver is the isolation of a rare population of cells for detailed analysis. For example, the vast majority of cells in the liver are hepatocytes (parenchymal fraction), which are significantly larger than non-parenchymal cells. By enriching the specific cellular compartments of the liver (i.e. parenchymal and non-parenchymal fractions), and selecting for CD45 negative cells, we are able to enrich the starting population of stem cells by over 600-fold.The proceduresdetailed in this report allow for a relatively rare population of cells from a solid organ to be sorted efficiently. This process can be utilized to isolateliver stem cells from normal murine liver as well as chronic liver injury models, which demonstrate increased liver stem cell proliferation. This method has clear advantages over standard immunohistochemistry of frozen or formalin fixed liver as functional studies using live cells can be performed after initial co-localization experiments. To accomplish the procedure outlined in this report, a working relationship with a research based flow-cytometry core is strongly encouraged as the details of FACS isolation are highly dependent on specialized instrumentation and a strong working knowledge of basic flow-cytometry procedures. The specific goal of this process is to isolate a population of liver stem cells that can be clonally expanded in vitro

Anion channel sensitivity to cytosolic organic acids implicates a central role for oxaloacetate in integrating ion flux with metabolism in stomatal guard cells

Stomatal guard cells play a key role in gas exchange for photosynthesis and in minimizing transpirational water loss from plants by opening and closing the stomatal pore. The bulk of the osmotic content driving stomatal movements depends on ionic fluxes across both the plasma membrane and tonoplast, the metabolism of organic acids, primarily Mal (Imitate), and its accumulation and loss. Anion channels at the plasma membrane are thought to comprise a major pathway for Mal efflux during stomatal closure, implicating their key role in linking solute flux with metabolism. Nonetheless, little is known of the regulation of anion channel current (I(Cl)) by cytosolic Mal or its immediate metabolite OAA (oxaloacetate). In the present study, we have examined the impact of Mal, OAA and of the monocarboxylic acid anion acetate in guard cells of Vicia faba L. and report that all three organic acids affect I(Cl), but with markedly different characteristics and sidedness to their activities. Most prominent was a suppression of I(Cl) by OAA within the physiological range of concentrations found in vivo. These findings indicate a capacity for OAA to co-ordinate organic acid metabolism with I(Cl), through the direct effect of organic acid pool size. The findings of the present study also add perspective to in vivo recordings using acetate-based electrolytes

Adaptive Management and the Value of Information: Learning Via Intervention in Epidemiology

Optimal intervention for disease outbreaks is often impeded by severe scientific uncertainty. Adaptive management (AM), long-used in natural resource management, is a structured decision-making approach to solving dynamic problems that accounts for the value of resolving uncertainty via real-time evaluation of alternative models. We propose an AM approach to design and evaluate intervention strategies in epidemiology, using real-time surveillance to resolve model uncertainty as management proceeds, with foot-and-mouth disease (FMD) culling and measles vaccination as case studies. We use simulations of alternative intervention strategies under competing models to quantify the effect of model uncertainty on decision making, in terms of the value of information, and quantify the benefit of adaptive versus static intervention strategies. Culling decisions during the 2001 UK FMD outbreak were contentious due to uncertainty about the spatial scale of transmission. The expected benefit of resolving this uncertainty prior to a new outbreak on a UK-like landscape would be £45–£60 million relative to the strategy that minimizes livestock losses averaged over alternate transmission models. AM during the outbreak would be expected to recover up to £20.1 million of this expected benefit. AM would also recommend a more conservative initial approach (culling of infected premises and dangerous contact farms) than would a fixed strategy (which would additionally require culling of contiguous premises). For optimal targeting of measles vaccination, based on an outbreak in Malawi in 2010, AM allows better distribution of resources across the affected region; its utility depends on uncertainty about both the at-risk population and logistical capacity. When daily vaccination rates are highly constrained, the optimal initial strategy is to conduct a small, quick campaign; a reduction in expected burden of approximately 10,000 cases could result if campaign targets can be updated on the basis of the true susceptible population. Formal incorporation of a policy to update future management actions in response to information gained in the course of an outbreak can change the optimal initial response and result in significant cost savings. AM provides a framework for using multiple models to facilitate public-health decision making and an objective basis for updating management actions in response to improved scientific understanding

Domestic Asset Protection Trusts: Ushering in the Klackaba Era

The growth in the U.S. economy has allowed Americans to increase their savings--but how? A novel approach has emerged in seventeen states: domestic asset product trusts (DAPTs). DAPTs are self-settled spindthrift trusts that allow the settlor to retain a beneficial interest in the trust while removing it from the reach of future creditors. Through the lens of the favorable ruling in Klackaba v. Nelson, this Note addresses why DAPTs should be regarded as an effective method of protecting a settlor’s money and argue for more states to follow suit