Bibliography: leaves 253-299. / 299 leaves : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Investigates the nature of disordered oesophageal motor function that may be of importance in the pathogenesis of gastro-oesophageal reflux disease. / Thesis (Ph.D.)--University of Adelaide, Dept. of Medicine, 1995

Hepatocellular carcinoma (HCC) is one of the most prevalent cancers worldwide. Hepatocarcinogenesis is considered a multifactorial and mulitstep process that involves the activation of oncogenes or the inactivation of tumor suppressor genes. Tumor suppressor gene PTEN (also known as MMAC or TEP1) is located on human chromosome 10q23. The 403¡Vamino acid PTEN protein encodes dual specificity protein phosphatases. Mutation of the PTEN is a common event in advanced stage of diverse human cancers. In our previous studies, immunohistochemical analysis indicated that reduced PTEN expression was found in nearly 40% of HCC specimens. Furthermore, restored PTEN expression by adenovirus gene delivery effectively inhibited the in vitro and in vivo tumorigenicity of Mahlavu cells, a human HCC cell line with PTEN inactivation. In the present study, we further characterize whether PTEN gene delivery still suppressed the oncogenic potential in HCC cell lines with functional PTEN. By expression and sequencing analysis, we identified human SK-Hep-1 cells as the hepatoma cell line with functional PTEN expression. The optimal condition for adenovirus vector to infect SK-Hep-1 cells was determined at the multiplicity of infection (MOI) of 50-100. Tough SK-Hep-1 cells were effectively transduced with exogenous PTEN gene, the enhanced PTEN expression by adenovirus gene delivery did not alter the phosphoryation extent of Akt in SK-Hep1 cells. Nevertheless, PTEN gene delivery reduced the proliferation of SK-Hep-1 cells by ~20%. In addition, the motility of PTEN-transduced SK-Hep-1 cells significantly decreased comparing to cells of control groups. Western blot analysis suggested the decreased cell motility might be attributed to the reduced phosphorylation of focal adhesion kinase (FAK) by PTEN gene delivery. Above all, PTEN gene delivery profoundly reduced the colony formation of SK-Hep-1 cells in soft-agar. However, PTEN gene delivery did not affect the secretion of matrix metallo-proteinases (MMPs) release. Animal studies will be carried out in the future to validate the present in vitro findings. In summary, PTEN gene delivery holds promise for treatment of HCC even when the hepatoma cells possess functional PTEN gene.

The goal of this research was to examine the role of the novel membrane progestin receptor alpha (mPR[alpha]) in the stimulation of sperm hypermotility by the progestin 17,20[beta],21-trihydroxy-4-pregnen-3-one (20[beta]-S) in two teleosts; the Atlantic croaker (Micropogonias undulatus) and the southern flounder (Platylicthys lethigstomata). In croaker, the expression, localization and hormonal regulation of mPR[alpha] in testis and sperm were investigated, as were the intracellular signaling pathways activated by 20[beta]-S and mPR[alpha] to induce croaker sperm hypermotility. In flounder, stimulation of sperm hypermotility by 20[beta]-S and binding of this steroid to flounder sperm membranes were examined. Finally, expression of mPR[alpha] was investigated in flounder testes and the expression and localization of this receptor in flounder testis and sperm was examined. In croaker sperm, mPR[alpha] was expressed on the plasma membrane and localized to the midpiece. Expression of mPR[alpha] was also shown to be associated with high sperm motility and regulated by gonadotropin. The signaling pathways activated by 20[beta]-S in croaker sperm were shown to involve activation of olfactory G-proteins (Golf). Subsequent activation of membrane adenylyl cyclases was also demonstrated and shown to be necessary for 20[beta]-S-stimulated cAMP production and 20[beta]-S-induction of sperm hypermotility. Furthermore, co-immunoprecipitation studies show mPR[alpha] and Golf physically associate with one another, establishing mPR[alpha] as the mediator of 20[beta]-S actions in croaker sperm. Finally, evidence was obtained for progestin-stimulation of sperm hypermotility and the presence of mPR[alpha] on sperm membranes in another marine teleost species belonging to a different family, the southern flounder. In addition, mPR[alpha] was shown to be expressed on flounder sperm membranes and also localized to the sperm midpiece. Results from the following studies support the hypothesis that mPR[alpha] is the mediator of 20[beta]S-stimulated sperm hypermotility in croaker and is a likely intermediary in southern flounder. Furthermore, these data provide a plausible mechanism by which 20[beta]-S and mPR[alpha] stimulate croaker sperm hypermotility. In addition, these results provide the first evidence of hormonal activation of Golf proteins for any species. Finally, mPR[alpha]-mediated mechanisms to increase sperm motility are suggested to be evolutionarily conserved in teleosts since they also likely exist in a non-sciaenid species, the southern flounder.

published_or_final_version / Zoology / Doctoral / Doctor of Philosophy

The role of supraspinal, spinal and peripheral alpha-2 adrenoceptors in the regulation of gastrointestinal motility in mice was investigated using anatomically site specific administration of clonidine and adrenoceptor antagonists. Clonidine produced a dose-dependent inhibition of gastrointestinal transit when given by the i.c.v., i.th., or s.c. routes, and was most potent when given i.c.v. Yohimbine, an alpha-2 adrenoceptor antagonist, but not the alpha-1 antagonist prazosin, antagonized the antitransit effects of clonidine. Yohimbine was most potent in antagonizing i.c.v. clonidine; increased doses of the i.c.v. antagonist were required when the agonist was given s.c. After transection of the spinal cord, i.th. clonidine failed to produce an antitransit effect. Additionally, the i.c.v. potency of clonidine decreased approximately 7-fold in spinally-transected mice. The data suggest that the antitransit effects of clonidine occur through actions at alpha-2 adrenoceptors located at both supraspinal and peripheral sites.

No description available.

<p>Gonadotropin Releasing Hormone (GnRH) is a hypothalmic decapeptide, which regulates mammalian gonadotropin secretions by binding to specific, high affinity receptors in the pituitary. Two forms of GnRH (GnRH I and GnRH II) are expressed in the brain of human and some primates. Even though primates have been used extensively in a variety of investigations in relation to the role of GnRH in reproduction, there is no evidence of any research to investigate the direct effect of GnRH on primate sperm.</p>

The variations in ER structure and motility under different stages of cell development remain largely unexplored. Here, I observe ER structure and the changes that take place in this structure over time in growing and non-growing live epidermal cells of the pea tendril. The ER was labelled by green fluorescent protein, fused to the KDEL-ER retention signal and confocal scanning laser microscopy was used to localize the fluorescent signal. I found both the structure and motility of growing cells to be different to non-growing cells. The growing cells had a more open arrangement of the cortical ER, fewer lamellae and showed greater tubular dynamics, while the non-growing cells had a denser arrangement of the cortical ER network, with more lamellae and less tubular dynamics. Furthermore, these differences in the cortical ER structure and dynamics were due to growth as, the ER in non-growing cells showed characteristics similar to those seen in growing cells when these cells were induced to grow by the exogenous application of auxin. These changes in ER structure and dynamics were dependant on both the microtubules and actin cytoskeleton networks.

Nitric oxide, a short-lived free radical and neurotransmitter, is responsible for decreased smooth muscle contractility in vitro. When in excess, NO can cause hypotension and is believed to mediate altered intestinal motility. Not enough evidence is available for morphological changes in gastrointestinal smooth muscle and its correlation with motility disorders caused by Escherichia coli-induced NO production. Male Lewis rats were treated with injections of 10 mg/kg LPS from E. coli with or without 12.5 mg/kg of NOS inhibitor, LNMMA. Eighteen to 24 hours following injection, duodenum, ileum, colon, liver, and spleen were harvested for histological analysis. Urine and fecal analysis assessed functional aspects in control and treatment groups. Muscularis externa measurements revealed significant increase in muscle thickness of LPS + LNMMA injected group compared to control and LPS group. However, the average values in control and LPS group were not significantly different. Fecal consistency was significant in all 3 groups. Mean urinary nitrite in the LPS group was 44 times higher than control and 52 times higher than the inhibitor-treated group. / Department of Physiology and Health Science

No description available.