An earlier approach to restore homeostatic levels of ECM degrading matrix metalloproteases (MMPs) by the Sefton Lab utilized hydroxamate-based MMP inhibitory (MI) beads. While the MI beads delayed ECM degradation in the context of skin wound healing, they caused elevated cell infiltration in a subcutaneous implant model. The primary goal of this project was to further investigate this finding using an air pouch implant model in mice and a different control group – methacrylic acid-based (MAA) beads. Exudate analysis indicated that the MI beads, implanted subcutaneously with gelatin discs, elicited a similar biological response as the MAA beads. Exudates corresponding to both biomaterials had similar cell counts and chemokine levels, which were greater than those corresponding to the control used earlier, poly-methyl methacrylate-based (PMMA) beads. Further, both MI and MAA beads activated infiltrating macrophages in the classical manner, and influenced the activity of an MMP8 catalytic domain in a similar manner.

The commensal intestinal microbiota contributes substantially to intestinal development in the early neonatal period by mechanisms that are not yet elucidated but could contribute to novel strategies to improve intestinal health. A series of gnotobiotic experiments using isolator-reared caesarian section-derived piglets inoculated at 1 d of age with selected bacteria and euthanized at 14 or 15 days of age were performed to investigate intestinal morphology, inflammation and digestive function. In Experiment 1, piglets were maintained germfree (GF), mono-associated with Escherichia coli (EC), mono-associated with Lactobacillus fermentum (LF) or conventionalized with sow feces (CV). Increased (P<0.05) gene expression of Fas ligand (FasL) and tumor necrosis factor (TNF?) in EC and CV as compared to LF and GF pigs coincided with increased apoptotic and proliferative activity. Toll-like receptors (TLR) 2, 4 and 9 were differentially regulated (P<0.05) by colonizing species. In Experiment 2 using the same animals as Exp. 1, increased turnover of brush border enzymes was indicated by reduced (P<0.05) specific activity of aminopeptidase N (APN) and lactase (LPH) and increased expression of APN in CV and EC as compared to GF and LF pigs. Reduced enzyme activity to gene expression ratio corresponded with an in vitro assay of microbial inactivation of APN. In Experiment 3, probiotic Lactobacillus sp., L3777, and Bifidobacteria sp., B5445, did not induce expression of inflammatory cytokines in mono-association but di-association with E. coli increased (P<0.05) inflammatory and anti-inflammatory mediators and resulted in a high rate of sepsis (50%) relative to E. coli mono-association. Induced expression of inflammatory cytokines by commensal bacteria through TLR and other means, appear to play a substantial role in microbially-induced enterocyte turnover. Enterocyte immaturity did not account for reduced enzyme activity associated with inflammation as increased expression of APN in response to microbial colonization was observed, suggesting a host response pathway enabling effective competition with the intestinal microbiota for available peptide nutrients. Probiotic bacteria were relatively benign in mono-association but may have facilitated increased translocation of <i>E. coli</i> in di-association. Gnotobiotic animal models are essential to demonstrate outcomes of host response characterized by communication among numerous cell types, although are of significant technical difficulty.

Dessertation describes of the first reported syntheses of the natural products lairdinol A and depsilairdin. The key steps in the synthesis of depsilairdin were N-terminal extension (C←N) of the protected proline fragment, hydrolysis of the tetrapeptide fragment with free secondary alcohol in the proline moiety and esterification of the HOBt ester of tetrapeptide fragment with the bromomagnesium alkoxide of lairdinol A.

Entry of herpesviruses is mediated by the interactions between viral glycoproteins and cellular receptors. Among these glycoproteins, gB plays an important role. In this study, my major focus was to study gB's functions in the virus entry process and the structural requirements for gB to conduct its functions. The virus model in my study is bovine herpesvirus 1 (BHV-1), a member of the alphaherpesviruses. BHV-1 gB is a type I integral membrane protein with a potential transmembrane anchor at the C-terminal region. A cleavage site in the middle divides this molecule into two subunits, gBb and gBc. In this study, a truncated gB, gBt (residues 1 to 763), and N-terminal subunit, gBb (residues 1 to 505), were first expressed under the control of the bovine heat-shock protein 70A (hsp70A) gene promoter in stably transfected Madin Darby bovine kidney (MDBK) cells. Both forms of gB were secreted into the medium with apparent molecular weights as anticipated, and they were reactive to all gB-specific monoclonal antibodies used in this study. Affinity-purified gBt and gBb were able to elicit antibody responses in mice to an extent comparable to those induced by authentic gB. These results suggest that gBt and gBb retain the structural and antigenic properties of authentic gB. Furthermore, the intracellular processing of gBt and gBb was similar to that of authentic gB in virus-infected cells. Finally, gBt was proteolytically cleaved after conversion of the high mannose-containing precursor to the mature form. These truncated gBs that were prepared served as reagents for the core of my studies. BHV-I gB can bind to heparin sulfate (HS) and another non-HS receptor on MDBK cells. We assume that high-affinity binding to the non-HS receptor is important for BHV-1 infectivity. BHV-1 gB forms dimers in infected cells and in virions, and its dimerization domain may be located between residues 506 to 763. The cytoplasmic domain of BHV-1 gB is important for the existence of the high-affinity binding site. Without the cytoplasmic domain, the truncated gB derivatives exhibit conformational changes and loss of the high-affinity binding site. By comparing the expression of different gB derivatives in MDBK cells, it was found that in the putative transmembrane region, segment 3 is the real membrane anchor, whereas segment 2 is the fusogenic domain. (Abstract shortened by UMI.)

The commensal intestinal microbiota contributes substantially to intestinal development in the early neonatal period by mechanisms that are not yet elucidated but could contribute to novel strategies to improve intestinal health. A series of gnotobiotic experiments using isolator-reared caesarian section-derived piglets inoculated at 1 d of age with selected bacteria and euthanized at 14 or 15 days of age were performed to investigate intestinal morphology, inflammation and digestive function. In Experiment 1, piglets were maintained germfree (GF), mono-associated with Escherichia coli (EC), mono-associated with Lactobacillus fermentum (LF) or conventionalized with sow feces (CV). Increased (P<0.05) gene expression of Fas ligand (FasL) and tumor necrosis factor (TNF?) in EC and CV as compared to LF and GF pigs coincided with increased apoptotic and proliferative activity. Toll-like receptors (TLR) 2, 4 and 9 were differentially regulated (P<0.05) by colonizing species. In Experiment 2 using the same animals as Exp. 1, increased turnover of brush border enzymes was indicated by reduced (P<0.05) specific activity of aminopeptidase N (APN) and lactase (LPH) and increased expression of APN in CV and EC as compared to GF and LF pigs. Reduced enzyme activity to gene expression ratio corresponded with an in vitro assay of microbial inactivation of APN. In Experiment 3, probiotic Lactobacillus sp., L3777, and Bifidobacteria sp., B5445, did not induce expression of inflammatory cytokines in mono-association but di-association with E. coli increased (P<0.05) inflammatory and anti-inflammatory mediators and resulted in a high rate of sepsis (50%) relative to E. coli mono-association. Induced expression of inflammatory cytokines by commensal bacteria through TLR and other means, appear to play a substantial role in microbially-induced enterocyte turnover. Enterocyte immaturity did not account for reduced enzyme activity associated with inflammation as increased expression of APN in response to microbial colonization was observed, suggesting a host response pathway enabling effective competition with the intestinal microbiota for available peptide nutrients. Probiotic bacteria were relatively benign in mono-association but may have facilitated increased translocation of <i>E. coli</i> in di-association. Gnotobiotic animal models are essential to demonstrate outcomes of host response characterized by communication among numerous cell types, although are of significant technical difficulty.

Dessertation describes of the first reported syntheses of the natural products lairdinol A and depsilairdin. The key steps in the synthesis of depsilairdin were N-terminal extension (C←N) of the protected proline fragment, hydrolysis of the tetrapeptide fragment with free secondary alcohol in the proline moiety and esterification of the HOBt ester of tetrapeptide fragment with the bromomagnesium alkoxide of lairdinol A.

With the rising prevalence of diabetes, effective means of successful management of blood glucose levels are increasingly important. To improve on the ease of measurements, new technology is being developed to enable less invasive measurements. Some recent efforts have focused on the development of optical microscale glucose sensing systems based on the encapsulation of glucose oxidase within microspheres coated with polyelectrolyte multilayer nanofilms. In such sensors, a phosphorescent oxygen indicator is also co-encapsulated with the enzyme inside so that when glucose is present, glucose oxidase within the sensor reduces the local oxygen levels, causing a corresponding change in the luminescence intensity of the sensors. To test the aforementioned factors, a two-substrate, 2D FEM model of microscale optical glucose sensors in the dermis was developed. The model was used to predict the response time and sensitivity of glucose sensors with varying number and spacing of particles distributed in the dermis and varying physiological characteristics of the surrounding tissue; specifically, capillary density, blood vessel location relative to sensor, and glucose and oxygen consumption in tissue. Simulations were conducted to determine the magnitude of the change in the response time of sensors. Because the steady-state oxygen concentration within the sensors for a given blood glucose level determines the signal output, steady-state concentration of oxygen within sensors and the surrounding tissue for the entire physiological glucose range was evaluated. The utility of the model to predict the performance and efficacy of the sensors in the event of a host response to the foreign body implant was also evaluated. Simulations were performed to evaluate changes in sensor response and sensitivity in the occurrence of inflammation and progression of fibrous encapsulation of various thickness and density. The results from these simulations have provided knowledge on the impact of physiological factors that can potentially degrade sensor function in vivo. Our results indicate that upon the occurrence of a host response, sensitivity is reduced while range is extended. Furthermore, using the model we have been able to determine which conditions in vivo improve response time, sensitivity, and the linear response range for these sensors.

Sex-change (or sequential hermaphroditism) is a phenomenon whereby an organism functions first as one sex, and then as the other in the later stage of its life. Sex change is an adaptation that has evolved to allow certain organisms to increase fitness through resource allocation of male and female functions in a single lifespan. Since the timing of sex change in an organism is critical to its reproductive output, the size or age at sex change is expected to be plastic in response to different environmental conditions. Social and environmental conditions usually convey the cues about the relative advantages of functioning as one sex or the other. Identifying the factors that influence the timing at sex change is important to understand the evolution of sex change in the life history. In order to elucidate how to optimize fitness through sex allocation in response to different environment, I studied the timing at sex change of Coralliophila violacea in different approach, including the age of individual, the compositions of social groups and the difference between two types of coral hosts in this thesis. Firstly, I present a method to determine the age of this snail and its age at sex change. The growth striae on the operculum were studied and compared to the age estimated by the Gompertz growth function based on growth data obtained from mark-recapture experiments. There is a significant correlation between the number of striae on the operculum and the age estimated from the Gompertz growth function, and the relationship is 1:1. These results suggest that the number of striae on the operculum can be used as an age index, with each stria representing 1 year of age. The age of sex change of this snail, according to my estimates by both stria number and aperture-length inferences, occurs between 4 and 6 years old. Growth rates of the snails are negatively correlated to size. Furthermore, individuals undergoing sex-change grew faster than males and females. This snail, Coralliophila violacea, forms snail patches on the surface of the massive coral, Porites spp. Secondly, the social factors were analyzed for the timing of this sedentary snail. These factors include the sex ratio, number of male and females, and the size of females in the snail patches, all of which could be important information in determining the timing of sex change. In this study only the size of the largest female could be significantly correlated with the size at sex change in a snail patch. Finally, I survey the distribution, size and reproductive characteristics of the snail, Coralliophila violacea (Lamarck), which inhabits the surface of both the branching coral Porites nigrescens and the massive corals, P. lobata and P. lutea. Based on these results I examined the host effect on (1) population structure and (2) reproductive characteristics, including the size at sex change of symbionts. On branching hosts, most snails were solitary, whereas on massive hosts, most had formed multiple-snail patches. Significantly smaller snails as well as proportionally more females were found on branching than on massive hosts. Furthermore, the fecundity of the females on the branching hosts was significantly lower than that on the massive hosts. The size at sex change (male to female) of the snails was smaller on the branching hosts than on massive hosts. Patch composition differences can partly explain the smaller size at sex change for snails on branching hosts; however, there was also evidence that host morphology had a significant effect on the timing of sex change.

This project focused on improving the in vitro rearing methods of Toxoneuron nigriceps, an important biological control agent of Heliothis virescens, by supplementing the rearing media with the host hemolymph. T. nigriceps larvae of different ages were incubated in the artificial rearing media. The growth (increase in length and width), development (molting), and survival of the incubated larvae were observed. Changes in osmotic pressure of the rearing media before and after incubation were evaluated. The protein profiles of day 1, 3 and 5 hemolymph samples of the parasitized and unparasitized H. virescens fifth instar larvae, used in the artificial rearing media, were determined. Host hemolymph improved the growth and development of T. nigriceps larvae in the artificial rearing media except in the case of new and early first instar larvae. Osmotic pressure of all media showed a significant decrease after the incubation of the larvae except media incubated with the new first instar larvae indicating that the older larvae absorbed and utilized more of the nutrients from the rearing media than the younger larvae. Growth and development was higher in the semisolid media than in the liquid media. In the artificial rearing media, the late first instar T. nigriceps larvae molted to second instars and a few of them molted to third instars. The early and late second instar larvae incubated in the artificial media also grew well and molted to third instars. Some of the in vitro reared third instar larvae demonstrated behavioral changes that could be expressed as the preparation for cocoon formation or pupation, however neither a cocoon nor pupation occurred. No qualitative differences in the protein titers were detected between hemolymph of the parasitized and unparasitized fifth instar H. virescens larvae. There was a significant increase in the protein concentration in both unparasitized and parasitized H. virescens larvae as the age increased. This increase in the protein concentration showed a positive effect on the growth and development of T. nigriceps larvae indicating that availability of nutrients was an important factor for the growth of T. nigriceps larvae in vitro.

The objectives of this dissertation are to examine how expatriates¡¦ social network affect the ability of access information/resource and knowing- whom capital on their career advancement, as well as the effects of perceived organizational support correlate to host and home social network. Specially, this research investigates dual sources (host and home office) of expatriates¡¦ network in order to understand the impact on the network abilities and extent. Through confirmed factor analysis of pretest data, measurements were refined and the questionnaire used for the study was developed. For the study, 267 surveys were used for the data analyses. Structural equation modeling in LISREL 8.8 was employed to analyze the relationships among the latent constructs. The findings from the study revealed significant relationships between 1) expatriates¡¦ social network of host office and access information & resource, 2) social network of host office and knowing whom capital, 3) expatriates¡¦ social network of home office and access information & resource, 4) access information & resource and career advancement, and knowing whom capital and career advancement. This study contribute theoretically to the expatriates dual social network literature by clarifying the relationships among host social network, home social network, social network abilities and career advancement, while the previous studies focus on expatriates host country context. This research findings pointed out the expatriates not re-establish a personal network in the host environment but still keep contact with his/her home social network. The expatriates¡¦ host social network has direct and strong effect on access to information/resources and knowing-whom capital. And the expatriates¡¦ home social network also has positive impact on access to information/resources. This empirical model also provides future research with a new angle of view over expatriates social network and career advancement.