Objective: This study was conducted to determine the effectiveness of physical and mechanical disinfection of P. gingivalis from implant disks and to evaluate bone cells growth and attachment to the disks. Background. Each year, over three million of Americans replacing missing teeth with dental implants. An inflammatory process around an implant that causes bone loss, characterizes peri-implantitis, first diagnosed in the 1980s. The prevalence is approximately 22%. To date, no treatment protocol of peri-implantitis has been proposed. Methods. 207 implants disks. Four different implant surfaces utilized. Disks were contaminated by p. gingivalis and consequentially disinfected by physical means (spraying prophy jet, titanium brush, and ultrasonic activation) and chemically by Hydrogen Peroxide 3%, 0.12% Chlorhexidine Gluconate, and Sodium Bicarbonate. Osteoblasts were added to the disks. Growth factors (Emdogain and Gem21S) were used in two groups. Osteoblast vitality, attachment and morphology were evaluated. Results. On 3iT3 the all disinfection methods had similar results. On Osseotite and Nanotite surfaces, the citric acid combined with ultrasonic activation granted the worse results. Hence, disks that did not have the surface altered by physical decontamination had most cells attached. Hydrogen Peroxide 3% showed to be the most biocompatible and 0.12% Chlorhexidine gluconate showed most cellular toxicity. Implant coating did not influence osteoblast attachment. Growth factors did not promote osteoblast attachment. Conclusion: Further investigations are necessary.

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第19746号 / 工博第4201号 / 新制||工||1648(附属図書館) / 32782 / 京都大学大学院工学研究科高分子化学専攻 / (主査)教授 田畑 泰彦, 教授 秋吉 一成, 教授 木村 俊作 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

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Indiana University-Purdue University Indianapolis (IUPUI) / CD1d is a cell surface glycolipid that, like Major Histocompatibility Complex (MHC) class I and MHC class II molecules, presents antigen. However, instead of peptides, CD1d presents lipids to Natural Killer (NK) T cells, a subset of T cells that express both NK cell markers and the T cell receptor and produces both T helper (Th) 1 and Th2 cytokines. Our lab focuses on the regulation CD1d-mediated antigen presentation. TGF beta is a known regulator of the immune system, such as controlling MHC class II antigen presentation. Further, TGF beta can activate the mitogen activated protein kinase (MAPK) p38, a known negative regulator of CD1d-mediated antigen presentation. Therefore, we hypothesized that TGF beta would be a negative regulator of CD1d-mediated antigen presentation, and our results showed a decrease in antigen presentation by CD1d in response to TGF beta treatment. However, this inhibition was not through p38 activation, as indicated by the absence of a rescue of CD1d-mediated antigen presentation in, TGF beta-treated, p38 dominant negative-expressing cells. Alternatively, the Smad pathway, the canonical pathway activated by TGF beta, was investigated through a lentivirus shRNA-mediated knockdown of Smad2, Smad3 and Smad4 proteins. Smad2 shRNA-expressing cells showed in an increase in CD1d-mediated antigen presentation, suggesting an inhibitory role for Smad2. In contrast, Smad3 shRNA-expressing cells did not differ from control cells. However, as in the case of Smad2, CD1d+ cells in which Smad4 was knocked down, were substantially better at CD1d-mediated antigen presentation than control cells, suggesting that it also negatively regulates antigen presentation. Overall, these studies demonstrate that the canonical TGF beta/Smad pathway regulates an important part of the host’s innate immune response, vis-à-vis CD1d-mediated antigen presentation.

Bone morphogenetic protein 2 (BMP2) is a secreted growth factor that is essential for proper embryonic development and proliferation. Our laboratory discovered a nuclear variant of BMP2 (nBMP2) which is produced when translation is initiated at an alternative start codon within the BMP2 gene. When translation occurs at the downstream start codon, the resulting protein lacks the ER signal peptide, thereby allowing cytoplasmic translation and nuclear localization. Our aim is to distinguish the role of this nuclear localized variant from secreted BMP2. Overexpression of nBMP2 in HEK293 and HT29 cell lines resulted in a higher percentage of cells in proliferative phases of the cell cycle. We determined that nBMP2 does not regulate cell cycle progression by inducing hyperphosphorylation of retinoblastoma protein (Rb), but it may regulate the cell cycle by interacting with ROC1. In order to examine the role of nBMP2 in vivo, we have generated a mouse model in which a mutation of the nuclear localization signal (NLS) disrupts nuclear localization of nBMP2. Aberrant crypts were more abundant in nBmp2NLStm azoxymethane (AOM) treated mice than in wild type mice. Furthermore, H&E staining of colonic tissue showed that mutant mice have increased levels of dysplasia and aberrant crypt foci. This work suggests that nBMP2 is involved in regulating cell cycle progression and proliferation, and therefore may play a role in tumorigenesis.

Bakgrund: Budgetering är ett fenomen som har använts i århundraden inom offentlig förvaltning där privata företag började använda detta verktyg betydligt senare. Budgetering har länge varit en del av verksamhetsstyrningen inom olika former av företag, där budgetens roll inom tillväxtbolag som verkar i föränderliga miljöer erhållit kritik. Dessa företag växer kraftigt där det ställs ett högre krav på budgetens aktualitet samtidigt som budgetering är en tids- och resurskrävande process som enligt kritiken ger låg avkastning. Planering och samordning har varit genomgående viktiga faktorer för organisationer gällande budgetering som varit avgörande för att lyckas nå sina mål och utveckla organisationen. Samtidigt som den kraftiga tillväxten gjort budgeten som styrverktyg allt mer kritiserat ställdes krav för att kunna hantera en snabbare utveckling i relation till den tidigare synen på budgeten som styrverktyg.   Syfte: Genom att studera budgetstyrning i ett tillväxtbolag är uppsatsens syfte att undersöka och förstå hur budgeten kan användas i ett tillväxtbolag som är verksamma i en föränderlig miljö samt hur processen ser ut och därmed vilken roll budgeten antar i ett tillväxtbolag. Det kommer att genomföras genom att jämföra tidigare teori och hur den kan kopplas och ställas i relation till ett företag med stark tillväxt.   Metod: Uppsatsen är en kvalitativ intervjustudie på ett tillväxtbolag. Empirin utgörs av tre stycken semi-strukturerade intervjuer med ett företag som är verksamma inom IT-branschen. Det empiriska materialet har även kompletterats av skriftliga svar via mailkontakt. Det empiriska materialet har sedan analyserats för att urskilja likheter och skillnader med den valda teoretiska referensramen för att kunna besvara uppsatsens frågeställning.    Slutsats: Studien visar att det studerade tillväxtbolaget arbetar med en form av rullande forecasts vid ett flertal tillfällen under ett verksamhetsår som säkerställer att man arbetar på rätt sätt och är på rätt väg för att uppnå de uppsatta tillväxtmålen som främst mäts kring försäljning, rörelseresultat och bruttovinst. Det är en form av uppdaterad budget i relation till den som arbetades fram vid verksamhetsårets början, vars aktualitet är högre. Budgeten används fortfarande som styrverktyg men det finns en strävan efter ett bättre lämpat styrverktyg för tillväxtbolag som är verksamma inom föränderliga miljöer

The development of new multiphoton absorbing materials has attracted the attention of researchers for the last two decades. The advantages that multiphoton absorbing materials offer, versus their one-photon absorbing counterparts, rely on the nature of the nonlinearity of the absorption process, where two photons are absorbed simultaneously offering increased 3D resolution, deeper penetration, and less photobleaching and photodamage as a result of a more confined excitation. The applications of efficient two-photon absorbing materials have been extensively expanding into the fields of photodynamic therapy, microscopy, and optical data storage. One of the fields where an increased interest in multiphoton absorbing materials has been most evident is in bioimaging, in particular, when different cellular processes and organelles need to be studied by fluorescence microscopy. The goal of this research was to develop efficient two-photon absorption (2PA) compounds to be used in fluorescence bioimaging, meaning that such compounds need to posses good optical properties, such as high fluorescence quantum yield, 2PA cross section, and photostability. In the first chapter of this dissertation, we describe the synthesis and structural characterization of a new series of fluorescent donor–acceptor and acceptor-acceptor molecules based on the fluorenyl ring system that incorporated functionalities such as alkynes and thiophene rings, through efficient Pd-catalyzed Sonogashira and Stille coupling reactions, in order to increase the length of the conjugation in our systems. These new molecules proved to have high two-photon absorption (2PA), and the effect of these functionalities on their 2PA cross section values was evaluated. Finally, their use in two-photon fluorescence microscopy (2PFM) imaging was demonstrated. iii One of the limitations of the compounds described in Chapter 1 was their poor water solubility; this issue was addressed in Chapter 2. The use of micelles in drug delivery has been shown to be an area of increasing interest over the last decade. In the bioimaging field, it is key to have dye molecules with a high degree of water solubility to enable cells to uptake the dye. By enclosing a hydrophobic dye in Pluronic® F-127 micelles, we developed a system that facilitates the use of 2PA molecules (typically hydrophobic) in biological systems for nonlinear biophotonic applications, specifically to image the lysosomes. Furthermore, we report in this chapter the efficient microwave-assisted synthesis of the dye used in this study. In addition, linear photophysical and photochemical parameters, two-photon absorption (2PA), and superfluorescence properties of the dye studied in Chapter 2, were investigated in Chapter 3. The steady-state absorption, fluorescence, and excitation anisotropy spectra of this dye were measured in several organic solvents and aqueous media. In Chapter 4, we describe the preparation and the use of an efficient and novel twophoton absorbing fluorescent probe conjugated to an antibody that confers selectivity towards the vascular endothelial growth factor receptor 2 (VEGFR-2) in porcine aortic endothelial cells that express this receptor (PAE-KDR). It is known that this receptor is overexpressed in certain cancer processes. Thus, targeting of this receptor will be useful to image the tumor vasculature. It was observed that when the dye was incubated with cells that do not express the receptor, no effective binding between the bioconjugate and the cells took place, resulting in very poor, nonspecific fluorescence images by both one and two-photon excitation. On the other hand, when the dye was incubated with cells that expressed VEGFR-2, efficient imaging of the cells was obtained, even at very low concentrations (0.4 μM). Moreover, incubation of the bioconjugate iv with tissue facilitated successful imaging of vasculature in mouse embryonic tissue

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