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Thesis (Ph. D.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. "August 2007" Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 1998. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 1998. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

In the present study, muscle growth and flesh quality have been investigated from both commercially farmed Atlantic halibut (Hippoglossus hippoglossus) (Aga marine AS, Norway)and halibut obtained from small-scale trials at Mørkvedbukta Research Station (Bodø University College, Norway). Morphometric techniques have been utilized to investigate fast muscle growth in halibut ranging from circa 2 g to 100 kg, and it was established that fast muscle fibre recruitment ceases when the fish attain approximately 81 and 177 cm, in the case of males and females, respectively. Different muscle fibre types were distinguished using histochemical (myosin ATPase and succinic dehydrogenase) and immunohistochemical (S-58, an antibody against slow muscle myosin) staining techniques. Females recruit twice as many fast muscle fibres compared to males, which allows them to reach a larger final size. Furthermore, the seasonal growth patterns during a one year production cycle in commercial farmed halibut revealed a winter depression in growth leading to loss of biomass, which was attributed to the maturation of males. Commercial farmed fish of equal size (~1.5 kg) showed sexual dimorphism of fast muscle fibre number, caused by a significantly higher rate of fast muscle fibre recruitment in females. During the winter season fast muscle fibres shrunk significantly, especially in male fish, as a consequence of loss of appetite, low water temperatures and sexual maturation. None of the female fish matured during the trial. Flesh quality of halibut deteriorated during winter and spring, since it had a softer appearance and significantly lower myotomal protein content, particularly in males. Cathepsin activity was measured using spectroscopy and showed a strong negative correlation to protein content, displaying a seasonal variation. The proteolytic depletion of fast muscle proteins affected the water holding capacity of the muscle (determined by centrifugation), which showed concomitant changes with the increase in cathepsin activity and drop in protein content. Despite the soft appearance, the firmness (shear force) of the flesh increased during the winter. The hydroxylysyl pyridinoline cross-link content of the collagen matrix, determined by HPLC, showed a strong correlation to the fillet texture. The increased firmness during the winter, a period of little (female) or negative growth (males), was probably due to an increased cross-linking of the collagen compartment. Partial sequences of IGF-I and IGF-II were cloned from fast muscle of Atlantic halibut, and their relative gene expression levels were determined along with those of cathepsin B, cathepsin D and IGF-IRa in male halibut using qPCR during a fasting and refeeding trial. Transcript levels of cathepsin B and to some extent cathepsin D were significantly higher during fasting than refeeding, suggesting an increased enzyme production during periods of food deprivation. A temporary increase in IGF-I transcripts was observed after 7 days refeeding suggesting that this growth factor is involved in muscle growth control. Both IGF- IRa and IGF-II were down-regulated during refeeding.

Beim Tissue Engineering ist die Vaskularisierung von größeren Zell-Matrix-Konstrukten nach Implantation bis heute ein großes Problem. Durch das initiale Fehlen eines mikrovaskulären Netzwerkes kommt es zu einem raschen Zellsterben im Scaffold. Aufgrund dessen war das Ziel dieser Arbeit, im Sinne des in situ-Tissue Engineering ein Scaffold auf Basis von mineralisiertem Kollagen zu entwickeln, welches mit dem angiogenen Wachstumsfaktor VEGF funktionalisiert wird, um den Prozess der Vaskularisierung – die Einsprossung von Blutgefäßen – zu fördern und gleichzeitig durch Chemoattraktion in vivo Zellen aus dem umliegenden Knochengewebe in das Innere des Scaffolds migrieren zu lassen, so dass eine beschleunigte Defektheilung erzielt wird. Poröse Scaffolds aus mineralisiertem Kollagen wurden durch zwei unterschiedliche Strategien funktionalisiert und durch in vitro-Testungen charakterisiert. Die erste Strategie umfasste die Heparin-Modifizierung der gesamten Scaffolds, während die zweite Strategie die Injizierung eines zentralen VEGF-haltiges Depots in das Scaffoldinnere darstellte. Neben der Charakterisierung der Scaffolds wurde die Freisetzungskinetik des Modellwachstumsfaktors VEGF aus den modifizierten Scaffolds untersucht und die biologische Aktivität des freigesetzten Faktors auf Endothelzellen getestet. Zusätzlich wurde bei der 2. Strategie, der Injizierung eines Wirkstoffdepots, die Ausbildung eines Wirkstoffgradienten und die zielgerichtete Migration von Endothelzellen in Richtung des Wirkstoffdepots analysiert.