Patient-Clinician Communications: A Design Intervention for Patients' Comprehension of Their Care

Master of Design in Integrative DesignUniversity of Michiganhttps://deepblue.lib.umich.edu/bitstream/2027.42/136866/1/JYShin_2017_MDes-Thesis.pd

Optimizing conditions to electroporate Rothia mucilaginosa

Rothia mucilaginosa (Rm) is a gram-positive bacterium residing in the oral cavity. Recent studies in our laboratory have shown that this microorganism is able to cleave gluten, including immunogenic domains implicated in celiac disease. This can be beneficial to patients with celiac disease because exploitation of Rm can provide a novel mode of treatment. The enzymes responsible for this cleavage are as yet unknown. The purpose of this study was to optimize the transformation efficiencies of Rm cells through electroporation, with the ultimate goal to create knock-out mutants for enzyme activity. We have determined various aspects of Rm cells relevant for this project: (1) the growth curve characteristics of Rm; (2) the presence of endogenous restriction enzyme activities; and (3) the conditions facilitating Rm electroporation by varying electroporation voltages. Furthermore, electroporation and transformation of the plasmid pUC18 was conducted in Escherichia coli. The growth curve of Rm cells in BHI growth medium incubated at 37°C while shaking showed a doubling time of approximately 3 hours in the logarithmic growth phase. Using a cell sonicate of Rm cells incubated with Lambda DNA and four different restriction enzyme buffers, we found that there were no apparent endogenous restriction enzyme activities detectable. For the electroporation experiments, we used previously published protocols for the bacterium Staphylococcus aureus, as a standard condition to electroporate Rm cells. Those studies have shown that changing electrical parameters during the electroporation would yield a high efficiency rate of gram-positive bacterial transformation (Lofblom et. al., 2006; Metzler et. al., 1992). Therefore in our study, we increased the field strengths (kV*cm-1) to electroporate Rm cells. Rm cells could not be successfully transformed, and we observed that field strengths exceeding 18 kV*cm-1 destroyed Rm cells. On the other hand, the transformation of E. coli with pUC18 was successful. Our studies have laid the groundwork for investigating the transformation of Rm cells, and future studies can use the results obtained to further investigate ways to optimize transformation of Rm cells for potential utility in celiac patients

Aminoguanidine inhibits aortic hydrogen peroxide production, VSMC NOX activity and hypercontractility in diabetic mice

<p>Abstract</p> <p>Background</p> <p>Dysfunctionally uncoupled endothelial nitric oxide synthase (eNOS) is involved in producing reactive oxygen species (ROS) in the diabetic endothelium. The present study investigated whether anti-diabetes drug Aminoguanidine (AG) has any effect on eNOS function and vascular oxidant stress.</p> <p>Methods and Results</p> <p>Blood glucose levels were increased to 452.0 ± 15.1 mg/dl in STZ-treated male C57BL/6J mice (148.4 ± 3.2 mg/dl in untreated controls). Aortic productions of NO^•and O₂^•-were measured specifically and sensitively using electron spin resonance. Diabetic mice had a marked increase in aortic O₂^•-production. Aortic hydrogen peroxide (H₂O₂) production was also increased in diabetic aortas and significantly attenuated by AG. AG however had only a marginal effect in reducing aortic O₂^•-production, which corresponded to a minimal effect in improving aortic nitric oxide (NO^•) bioavailability. The endothelium-dependent vasodilatation however was modestly but significantly improved by AG, likely consequent to AG-induced reduction in hyper-contractility. NAD(P)H oxidase (NOX)-dependent O₂^•-production was completely attenuated by AG in endothelium-denuded diabetic aortas.</p> <p>Conclusion</p> <p>In summary, despite that AG is not an effective eNOS recoupling agent presumably consequent to its ineffectiveness in preventing endothelial NOX activation, it is inhibitory of aortic H₂O₂production, VSMC NOX activity, and hypercontractility in diabetes.</p

A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications.

The visual detection of specific double-stranded DNA sequences possesses great potential for the development of diagnostics. Zinc finger domains provide a powerful scaffold for creating custom DNA-binding proteins that recognize specific DNA sequences. We previously demonstrated sequence-enabled reassembly of TEM-1 β-lactamase (SEER-LAC), a system consisting of two inactive fragments of β-lactamase each linked to engineered zinc finger proteins (ZFPs). Here the SEER-LAC system was applied to develop ZFP arrays that function as simple devices to identify bacterial double-stranded DNA sequences. The ZFP arrays provided a quantitative assay with a detection limit of 50 fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5 min. The ZFP arrays provided sufficient sensitivity and high specificity to recognize specific DNA sequences. These results suggest that ZFP arrays have the potential to be developed into a simple and rapid point-of-care (POC) diagnostic for the multiplexed detection of pathogens

Interaction of Apoptotic Cells with Macrophages Upregulates COX-2/PGE 2

Recognition of apoptotic cells by macrophages is crucial for resolution of inflammation, immune tolerance, and tissue repair. Cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2) and hepatocyte growth factor (HGF) play important roles in the tissue repair process. We investigated the characteristics of macrophage COX-2 and PGE2 expression mediated by apoptotic cells and then determined how macrophages exposed to apoptotic cells in vitro and in vivo orchestrate the interaction between COX-2/PGE2 and HGF signaling pathways. Exposure of RAW 264.7 cells and primary peritoneal macrophages to apoptotic cells resulted in induction of COX-2 and PGE2. The COX-2 inhibitor NS-398 suppressed apoptotic cell-induced PGE2 production. Both NS-398 and COX-2-siRNA, as well as the PGE2 receptor EP2 antagonist, blocked HGF expression in response to apoptotic cells. In addition, the HGF receptor antagonist suppressed increases in COX-2 and PGE2 induction. The in vivo relevance of the interaction between the COX-2/PGE2 and HGF pathways through a positive feedback loop was shown in cultured alveolar macrophages following in vivo exposure of bleomycin-stimulated lungs to apoptotic cells. Our results demonstrate that upregulation of the COX-2/PGE2 and HGF in macrophages following exposure to apoptotic cells represents a mechanism for mediating the anti-inflammatory and antifibrotic consequences of apoptotic cell recognition

Community of Strangers: Itaewon from 'Americanized' Ghetto to 'Multicultural' Space

Ph.DDOCTOR OF PHILOSOPH

An Empirical Investigation into the Effects of Selected Price Heuristics on Tourists’ Purchase Decisions in Three Different Cultures

The central goal of this research was to investigate whether selected price heuristics are culturally specific or universal. The dissertation’s three experiments explore selected price heuristics in the context of tourism services among samples from the U.S., Korea and China; the U.S. represents an individualist and low context culture, while Korea and China represent collectivist and high context cultures. Study 1 investigated potential tourists’ price decisions when confronted with inexpensive functional items (buying a sandwich and a pizza) and an expensive hedonic option (purchasing tickets for a show); in two consuming situations (consuming alone or with another person); in two social group contexts (with an acquaintance or with a family member). The collectivist cultures showed social groups and cultures had a significant impact on price decisions, although they differed in their reactions when the group was a family member or an acquaintance. Hence, these variables had mixed influence on the U.S. sample’s responses. Study 2 measured the extent to which 9-ending digits were used in prices by suppliers of five tourism services both within and across the three different cultures represented by New York City, Seoul, and Shanghai. 9- and 8-ending prices were dominant in New York City and in Shanghai, respectively, but these culture specific endings were complemented by the universality of the 0- and 5-digit endings of prices which were ranked first and second, respectively, in Seoul, second and third in Shanghai, and third and second in New York. Study 3 investigated the relative strength of symbolic meanings of 9-ending prices (i.e. low quality, enhanced value, discount price, and misleading action) among samples from the U.S., Korea and China, and their effectiveness in influencing tourists' purchases. The analyses found no differences among the three cultures’ samples in either their relative importance across cultures, or in the likelihood of tourists selecting 9-ending rather than even-ending prices when purchasing a sandwich, a pizza or show tickets. In the context of a hotel room, the 9-ending discount was perceived to be a greater discount than even-ended prices, but its effectiveness could not be explained by the different symbolic meanings associated with 9-ending prices