The field of analytical chemistry is very important to today's society as more and more regulations and legislations emerge regarding trace elements in food, consumer products, medicines, and the environment. Like many areas of science, the current goals of trace elemental measurements and speciation are to increase knowledge on the subject and to improve upon current techniques by enhancing the figures of merit, such as accuracy and reproducibility, meanwhile balancing with the cost and time of analysis. The topics covered in this work were investigated primarily through the use of inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma optical emission spectrometry (ICP-OES). The phenomenon of compound-dependent responses in plasma spectrometry is discussed, seeking possible causes of it and offering some advice on how to prevent it. A new method was developed for the speciation of selenium in dietary supplements using anion-exchange chromatography ICP-MS. A novel method for the determination of total mercury at trace concentrations in rice was developed for use with conventional ICP-MS. Inductively coupled plasma mass spectrometry was also used for fingerprinting the rare earth elements in Maya archaeological pottery for provenance studies.

For a lay summary of the thesis presented in a 1-minute video format, visit the following link: https://www.youtube.com/watch?v=VhLzt_tEz-s / It is projected that, by 2030, 8% of all adults in the world will have diabetes mellitus and treatment will account for 10% of the total healthcare budget in many countries. Polymeric biomaterial research has led to the design of robust polymer hydrogel capsules to develop curative cell-based therapies for chronic disorders such as diabetes mellitus. Encapsulation of insulin-producing beta cells within synthetic, semi-permeable polymer hydrogels can avoid host immune rejection including fibrotic responses, and thus holds the promise of a long-term curative treatment of this disease. There is a paucity of literature regarding methods available for standardized in vitro screening of synthetic polymer hydrogel capsules to predict host responses in vivo. Thus, the focus of this thesis was to design in vitro assays able to screen for subsequent in vivo fibrotic responses. Two dimensional (‘2D’) (cell attachment to thin film hydrogel coatings) and three dimensional (‘3D’) (cell attachment and protein adsorption to hydrogel capsules) in vitro experiments were designed and tested in an iterative process to assess fibrotic responses to a diverse group of polymer hydrogels. Cell attachment assays included fibroblast (NIH 3T3) and macrophage (RAW 264.7) cell lines, and protein adsorption assays included proteins used to model fibrosis including fibrinogen and lysozyme. For some formulations, in vitro assays were compared with in vivo data on pericapsular cellular overgrowth (PCO) after being implanted into mice. A binomial logistic regression model was designed and validated to assess whether the ‘3D’ in vitro assays correlated with in vivo PCO responses. It was found that the RAW 264.7 cell attachment assay was significantly correlated with PCO outcomes in vivo, demonstrating for the first time a simple, cost-effective, and rapid in vitro cell-based approach to screen and select capsules with lower fibrotic potential to be further tested in animals. / Thesis / Master of Health Sciences (MSc) / In North America, one in eleven adults, or about 415 million people, have diabetes. It is projected that by 2030, around 8% of the world population will be diagnosed with this disease. A common form of treatment is through the frequent injection of insulin, but this is costly, requires multiple daily interventions, and cannot prevent regular excursions from the ideal blood glucose range. Cell-based therapies have a lot of promise in treating several chronic diseases including diabetes. Donor and stem-cell derived islets can be implanted into patients with type 1 diabetes and have been shown to function for over a year, albeit at the price of systematic immune suppression. Alternatively, cells that produce insulin can be placed inside immune-evasive capsules and implanted, potentially providing continuous blood glucose regulation without the need for daily insulin injections. However, this novel form of treatment is limited by the encapsulated cells’ survival once implanted. Cell survival can be affected by the body’s response to a foreign body (the capsule), causing deposition of protein or cells on the capsule surface which can limit the oxygen supply to cells in the capsule and the ability of insulin to leave the capsule in a timely fashion. The goal of this project is to develop assays to screen new capsule formulations. This can advance research by using capsules more readily accepted by the body, leading to a more promising and long-term treatment of diabetes.

The Streptococcus Anginosus/Milleri Group (SMG) is made up of three closely related but distinct bacterial species: Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus. The SMG are recovered from about one-third of healthy, asymptomatic individuals. Despite this, the SMG cause more incidences of invasive streptococcal disease than Group A and Group B Streptococcus combined. Members of this group are somehow able to live a dual lifestyle. Little work has been conducted on the molecular pathogenicity of the SMG and host factors that contribute to host susceptibility to this group have been under-investigated. My research works towards discovering how the host recognizes the SMG as well as what enables the SMG to evade clearance by the immune system. I hypothesize that: 1) recognition of the SMG by toll-like receptor 2 (TLR2) plays a key role in triggering a cytokine response by the innate immune branch (which coordinates the immune response to the SMG), 2) the expression of cytolysins and extracellular polysaccharides by members of the SMG enables evasion of innate immune recognition and cytokine responses. hTLR2 reporter and monocyte-like cell lines as well as human blood samples from healthy donors were used to investigate the host factors that contribute to SMG infection. Five clinical reference SMG strains and a transposon mutant library were used to probe the contributing bacterial factors. It was found that TLR2 activation plays an important role in the cytokine response to the SMG, but there is heterogeneity between strains in their ability to activate TLR2. It was also found that intermedilysin expression by S. intermedius strains enables evasion of recognition; however, different hosts display varying susceptibility to this cytolysin. This study reveals that investigation of both host and microbial factors is essential to build an understanding of the mechanisms of SMG transition from commensalism to pathogenicity. / Thesis / Master of Science (MSc) / The Streptococcus Miller/Anginosus Group (SMG) is a group of bacteria comprised of three species. Members of this group are recovered from roughly one-third of healthy individuals. However, the SMG are also found in samples collected from patients with invasive disease. It is not well understood why some human-SMG relationships are pathogenic and others are not. However, it is likely that the combination of both human and SMG factors determine the nature of the relationship formed between the two. In this study, the human and SMG factors that contribute to infection were investigated. The ways by which human cells recognize members of the SMG and defend themselves from damage was explored. Additionally, SMG factors that potentially contribute to infection were probed to discover their effect on human cells. By investigating both the bacterial and host factors that lead to infection, disease treatments and preventative strategies can be tailored to individual cases.

京都大学 / 新制・課程博士 / 博士(理学) / 甲第24183号 / 理博第4874号 / 新制||理||1697(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)准教授 足立 幾磨, 准教授 Huffman Michael Alan, 教授 今井 啓雄 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DFAM

Data for this study were derived from videotapes of 26 father-infant dyads, specifically from a five minute period of free-play. The first step was the creation of a literal transcription of the father-infant dyads interaction. Subsequently, nine variables of fathers' language characteristics and one infant characteristic were coded employing the literal transcriptions and observing the videotapes. The fathers' language variables were number of : (1) father utterances, (2) father words, (3) father contingent responses, (4) father teaching utterances, (5) father descriptive teaching utterances, (6) father directive teaching utterances - making commands, (7) father directive teaching utterance - asking questions, (8) percentage of father teaching utterances, and (9) mean length of father utterances (MLU). The infant variable was number of vocalizations. Eight out of the nine variables were positively correlated to infant vocalizations, indicating the importance of fathers input in child language development. The only negative correlation in the present study was between Mean Length of Utterance (MLU) and infant vocalizations and the possible reasons are discussed. The findings support the idea that there are positive relationships between fathers' language characteristics and infant vocalizations. Recommendations are made that fathers should be involved in early intervention programs.

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Given the high number of errors and negative events committed within medical settings, the emphasis on patient safety culture is becoming more prevalent. Despite this effort, underreporting has been and continues to be an issue in this area. Some research has shown a link between underreporting and lack of management responsiveness, but more work is necessary to identify reasons for underreporting and potential mitigating solutions. The objective of the present research is to answer questions regarding the impact supervisors have on staff’s patient safety perceptions and event reporting, through the use of archival survey data collected with the AHRQ Hospital Safety Culture Survey (2004). Probable moderators and mediators of key relationships were explored as well. Results are presented and their implications are discussed herein.

Coastal restoration has become a necessary and ubiquitous practice to enhance and conserve the many ecosystem services lost by marsh degradation. Wave climate is one of the most critical factors to consider for restoration projects. However, knowledge of the ways that waves affect marsh plants and the ecosystem services they provide is limited. The purpose of my dissertation was to improve the effectiveness of coastal marsh restoration by addressing the limitations and gaps associated with plant and ecosystem responses to waves through empirical research with three primary goals: 1) develop and test a low-cost wave gauge, 2) use it to compare above- and below-ground plant growth responses along a wave climate gradient, and 3) evaluate the effects of waves on nutrient removal in constructed marshes. I used three field and laboratory experiments to accomplish these goals. The low-cost wave gauge was developed using an Arduino microcontroller and various accessories. After development, the gauge was evaluated against a commercial gauge in a series of laboratory and field tests. Comparisons revealed over 90% agreement between the gauges and confirmed the applicability of the low-cost gauge. A total of thirty gauges were then constructed and deployed at sites within Mobile Bay, Alabama and surrounding tributaries. In addition to wave energy, plant data was also collected at each site, including above- and below-ground biomass, shoot density, height, and diameter. These data suggested that waves affect plant growth responses in ways not explained by the current plant response paradigm. For example, while greater diameter shoots best attenuate waves, shoot diameter declined with greater wave exposure in this study. This response was common among the study species. Other plant responses were species-specific. Finally, a field experiment was constructed to examine the main and interactive effects of sediment type, initial planting density, platform slope, and platform position at sites exposed to and protected from waves. Results from this experiment suggested that waves may potentially mitigate the effective removal potential of constructed marshes. Taken together, this dissertation advances research on plant responses to waves and provides new tools for land managers working on coastal restoration and conservation projects.

Contaminated poultry and egg products are major sources of Salmonella enterica serovar Enteritidis (S. enteritidis, SE) infections in humans. Colonization of SE in chicken reproductive tract results in the production of contaminated commercial shell-eggs and fertilized hatchery eggs. The complex pathogen-host interactions during SE colonization of chicken reproductive tract are largely unknown. This study was aimed at determining the pathogenic roles of the type three secretion systems (TTSS-1 and TTSS-2) in SE infection of chicken oviduct epithelial cells (COEC). A series of SE strains carrying mutations in the genes encoding structure or effector proteins of TTSS-1 and TTSS-2 were constructed. The invasiveness and intracellular survival rate of each SE strain as well as the host innate immune responses induced by the infections were evaluated. The results demonstrate that both TTSS-1 and TTSS-2 are required by SE to invade COEC which involve genes encoding effector proteins SipA, SopB, SopE2, and PipB. In addition to their involvement in host cell invasion, sipA and sipB are also necessary for the survival or replication of SE inside COEC. Inactivation of TTSS-2 genes (ssaV and pipB) resulted in an enhanced bacterial proliferation inside COEC. The data from this study also show that SE infection triggers pro-inflammatory responses in COEC and TTSS-1 is involved in the expression of iNOS and IL-8, a CXC chemokine. TTSS-1 and TTSS-2 are not necessary for induction of K203, MIP-1β, and IL-10 or suppression of TGF-β3 in COEC.

No description available.