published_or_final_version / Microbiology / Doctoral / Doctor of Philosophy

Fertility Associated Antigen (FAA) produced by the accessory sex glands and contained within the seminal fluid binds heparin and facilitates capacitation in ejaculated sperm, resulting in improved fertility in bulls capable of producing the protein. In this study, a total of 206 bulls derived from three populations were evaluated for the presence or absence of FAA through utilization of the Repro Test at three semen collections over a 60-d period. Across all collections, the percentage of FAA Negative bulls ranged from 13.64 to 36.11%. Within the three populations, 32, 33, and 67 bulls were observed at three different collections, of which 3.03, 9.09 and 4.48% were FAA Negative at all three collections, respectively. Furthermore, 27.27, 33.33, and 20.90% of bulls were observed to have variations within their FAA status after providing an initial FAA Positive result, respectively. Bull age, sperm concentration, progressive forward motility, percent normal sperm, ejaculate volume, and scrotal circumference were determined to be significantly different between FAA Negative and FAA Positive bulls in at least one collection. However, no consistent trend was observed across populations, or collections within a population, with regard to a relationship between these variables and FAA. Furthermore, of fourteen bulls that produced an ejaculate in which no sperm was detected, 78.57% (n=11) were FAA Positive despite the lack of sperm within the ejaculate. No single variable commonly measured to determine bull fertility was consistent in predicting the FAA status of bulls. The ability to produce FAA precedes puberty and the Repro Test can be used to identify FAA in prepuberal bulls. However, a large percentage of bulls, both prepuberal and peripuberal, are capable of displaying variation in their FAA status (as determined by the Repro Test) over time.

Thesis (Ph. D.)--University of Adelaide, 1986. / Offprints of author's four journal articles in pocket. Includes bibliographical references (leaves 131-147).

Thesis (Ph. D.)--University of Hong Kong, 2000. / Includes bibliographical references (leaves 150-157).

Technologies for early detection of tumors is critical for better therapy outcome and overall change in cancer survival. These assays must be capable of detecting tumors at early stages in order to prevent metastasis of the tumor and help reduce mortality. Biological molecules can serve as markers that can indicate the presence of cancerous cells. Current biomarkers approved by the FDA include CA 125, which is a tumor associated antigen (TAA). However, the sensitivities of these TAAs is not high enough to detect at early stages of disease. Recent technologies have found that antibodies that recognize these TAAs, also known as autoantibodies, provide more sensitive means to screen for tumors. This review aims to present recent literature data relative to the field of cancer diagnosis and treatment. However, one should note that this article covers only fraction of the broad science behind this subject.

Offprints of author's four journal articles in pocket / Bibliography: leaves 131-147 / x, 148 leaves, [13] leaves of plates : ill. (1 col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, 1986

Members of the membrane-spanning four-domains, subfamily A (MS4A) family are small polypeptides that share the structural features of four-transmembrane domains and unevenly sized extracellular loops. The family includes CD20, FcεRIβ and HtM4, plus a number of relatively uncharacterised proteins / predicted proteins. MS4A proteins are discussed in relation to other protein families, such as the tetraspanins, that are also characterised by four-transmembrane domains. The aim of this study was to identify the cell and tissue distribution, subcellular localisation, and function of a newly discovered member of the MS4A family, hepatocellular carcinoma-associated antigen 112 (HCA112). At a subcellular level, HCA112 was found on the plasma membrane of transfected COS-7 cells, and also within the Golgi complex, trans-Golgi-network, and early endosomes. The molecule is orientated such that the large loop is extracellular and the Nand C-terminal domains are cytoplasmic. The presence of HCA112 associated with components of the endocytic pathway raised the question of whether some originated from the surface membrane. Antibody was used to label a HA epitope tag engineered into the large extracellular loop of HCA112, and the bound antibody was tracked through early endosomes to the recycling compartment. Here it co-localised with internalised transferrin, indicating strongly that HCA112 is internalised via clathrin-dependent mechanisms. Several endocytic sorting motifs within the intracellular domains of HCA112 were investigated for their ability to direct internalisation of HCA112. Deletion of a di-leucine motif was found to slow but not prevent endocytosis, suggesting that it is involved in endocytosis of HCA112, although not essential for the process. When HCA112 expression constructs featuring N- and C-terminal domain truncations were examined, it was found that the N-terminal tail does not affect the subcellular localisation or trafficking of HCA112, while deletion of the C-terminal intracellular domain resulted in retention of the mutant protein in the ER. HCA112 has a wide tissue distribution and is highly expressed in the lining/covering and parenchymal epithelium of some tissues, proximal renal tubules, ductal epithelium in a number of organs, endothelial cells, some steroidogenic endocrine cells, adipocytes, smooth muscle cells, follicular dendritic cells and macrophages. The expression of HCA112 by a wide range of cell types suggests that its function(s) has general importance and is not limited to any specific cell type(s). After reflection on the functions of the HCA112-expressing cells, a common theme that emerged was one of endocytic activity. This lead to speculate that one function of HCA112 might be related to uptake of macromolecules, for instance, in antigen processing and presentation. This might be a general function, such as facilitating uptake of other cell membrane proteins, or directing the traffic of endocytic vesicles. It was noted that HCA112 has a similar cell and tissue distribution to the scavenger receptor and fatty acid translocase FAT/CD36 (Zhang et al., 2003). Furthermore, in cells co-transfected with HCA112 and FAT/CD36, the two molecules co-localise in early endosomes and co-immunoprecipitate, suggesting that the molecules physically and spatially associate. Thus, HCA112 could be involved with (or complement) FAT/CD36 in its functions as a long chain fatty acid transporter and scavenger receptor. A proteomics study of proteins that co-immunoprecipitated with HCA112 detected putative interactions with a number of proteins. These included LR8, transferrin receptor, interferon induced transmembrane proteins 2 and 3, Calpain-6, stomatin, PDGF α receptor, and heat shock 70 kDa protein 8 (HSPA8, formerly known as clathrin un-coating ATPase). Of these, LR8 and the transferrin receptor were investigated in more detail. The results provide strong evidence that HCA112 forms a novel complex with LR8, and that this may be involved in macromolecule internalisation or trafficking of membrane proteins, such as FAT/CD36 or the transferrin receptor. In the case of the transferrin receptor, this traffic appears to involve the clathrin-dependent pathway, but it is possible that when HCA112 is associated with FAT/CD36, it functions within lipid raft domains. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1375454 / Thesis (Ph.D.) -- University of Adelaide, School of Molecular and Biomedical Science, 2009

Cancerassocierade antigen (CA) används vid diagnostisering, prognosbestämning och för att följa behandling vid cancer. Exempel på CA är CA 15–3, CA 19–9 och CA 125. Syftet med arbetet var att undersöka hållbarheten för blodprover förvarade i primärrör som skulle analyseras för CA 15–3, CA 19–9 och CA 125 och jämföra med nuvarande metod på klinisk kemi, Oskarshamn, Region Kalmar län. Enligt nuvarande metod överförs plasman till nya provrör innan analys. Metoden som användes var att dubbelprov togs och det ena förvarades i primärrör och analyserades på Roche Cobas 411® tre, fem och tio dagar efter provtagning. Det andra provet överfördes till nytt provrör inför analys. Den procentuella skillnaden mellan analys av CA i primärrör och överfört prov beräknades. För alla tre analyserna var den procentuella skillnaden störst hos de lägsta koncentrationerna av CA. Medelvärde för analyserna av CA, standardavvikelse (SD) och variationskoefficient beräknades. SD för CA jämfördes mot kontrollerna MAS Omni Immune Pro 1 och MAS Omni Immune Pro 3s SD. Provernas SD var mindre för kontrollerna förutom för CA 19–9 >1100 kIE/L. I den gruppen analyserades dock för få analyser för att utföra statistisk analys och gav därmed ingen slutsats hur CA 19–9 påverkades av de olika förvaringarna. För övriga analyser och för CA 19–9 i lägre koncentrationer kunde slutsatsen dras att prover kunde förvaras i primärrör i tio dagar utan att analysresultatet påverkades. / Cancer-associated antigens (CA) are used for diagnosis, establish prognosis and to monitor treatment of cancer. Three CA are CA 15-3, CA 19-9 and CA 125. The aim of this study was to evaluate the stability of CA 15-3, CA 19-9 and CA 125 blood samples in primary tubes coated with Litium-heparin gel and compare the results with the method used at Klinisk Kemi Oskarshamn, Region Kalmar where the samples today are transferred to new tubes before analysis. In this study two samples were collected from each patient, one was stored and analyzed according to the current method. The other sample was stored in the primary tube and analyzed three, five and ten days after collection. Roche Cobas 411 instrument was used for analysis. The mean value for the samples in primary tubes was compared to the current method and were calculated in percent. The difference was more distinct when the concentration of the analyte was low. The mean value, standard deviation (SD) and variation coefficient were calculated for each analyte. The SD of the samples were compared to the SD of the controls IM 1 and IM3. All the obtained SD from the samples were lower than the control SD, except for CA 19-9 > 1100 kIE/L. Unfortunately, there were only three test results in the concentration interval and therefore no statistic conclusion could be made. The conclusion for the other analytes CA 15-3, CA 125 and CA 19-9 in lower concentrations was that the storage in primary tubed up to ten days did not affect the test results.

Orientador: Hélio José Montassier / Banca: Aramis Augusto Pinto / Banca: José Antonio Jerez / Resumo: A pesquisa sorológica usando Imunodifusão em Gel de Ágar (IOGA) e ELISA Bloqueio de Fase Líquida (BFL-EUSA) foram aplicadas nesse estudo no período de 1996 a 1998, para detectar ou titular, respectivamente, anticorpos contra o antígeno VIA (Virus Infection Associated) e as estirpes de referência 01 Campos, A24 Cruzeiro e C3 Indaial, em algumas espécies silvestres e domésticas susceptíveis e não vacinadas. Do total de 383 amostras sanguíneas colhidas, 78 amostras eram de bovinos de vida livre; 39 de búfalos indianos; 139 de ovinos; 63 de suínos domésticos; 49 de porcos do mato e 15 de cervídeos. Todas as espécies investigadas, exceto os cervídeos, apresentaram anticorpos contra o Vírus da Febre Aftosa (VFA). O teste de IDGA detectou 47 (12,3%) amostras positivas ao antígeno VIA, principalmente em bovinos baguás e búfalos. O BFL-ELISA identificou 175 (45,7%) soros com presença de anticorpos a uma das três estirpes virais; Foi encontrado um maior número de amostras positivas contra a estirpe C3 Indaial bem como maiores títulos, em bovinos baguás. Os bubalinos e ovinos apresentaram uma maior reatividade contra as estirpes A24 Cruzeiro e 01 Campos, respectivamente, demonstrando maior número de soros reagentes. No entanto, ambas as espécies revelaram maiores títulos para a estirpe C3 Indaial. Nos suínos domésticos e silvestres, predominou a reatividade a estirpe A24 Cruzeiro. Em bovinos, a maior ocorrência de soros positivos ao teste de IOGA foi encontrada em animais entre 12 a 36 meses de idade, e na prova BFL-ELISA, em animais acima de 36 meses. Nas demais espécies, ambos os testes laboratoriais utilizados demonstraram um maior número de soros reativos em indivíduos adultos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: A serologic survey, using Agar Gel Immunodifusion (AGID) and liquid Phase Blocking-ELISA (LPB-ELISA) were assessed in this study, in the period from to 1996 to 1998, to detect or titer, respectively, antibodies to Virus Infection Associated Antigen (VIAA) and the foot-and-mouth disease virus (FMDV) 01 Campos, A24 Cruzeiro e C3 Indaial reference strains, in non-vaccinated domestic and wild susceptible animal species. A total of 383 blood serum samples were collected from free-ranging cattle (78 samples), water buffalo (39 samples), sheep (139 samples), domestic (63 samples) and wild pigs (49 samples) and deers (15 samples), living in the Pantanal (Nhecolândia region) of Mato Grosso do Sul State, Brazil. All the investigated species, excepting deer, showed positive animais to FMDV antigens. The AGID detected a total of 47 (12,3%) positive animais for VIA antigen, mostly of them were free-living cattle and buffalo. The LPD-EUSA identified 175 (45,7%) serum samples as reactive to FMDV antigens, specially among cattle, buffalo and sheep. The predominant reactivity to FMDV antigens, characterized by the higher number of positives and the higher antibodies titers, were detected to C3 Indaial strain, particularly among the free-Iiving cattle. Buffaloes and pigs showed, in turn, a prominent reactivity to A24 Cruzeiro strain, while sheep reacted more to 01 Campos strain, though buffalo and sheep serum samples had higher antibody titers to C3 Indaial strain. The highest frequency of positive animais to VIA antigen were found in free-ranging cattle, whose age ranged from 12 to 36 months, contrasting to the results of LPB-EUSA that detected the highest rate of positives in the older bovines (~36 months). The older animais from other species demonstrated, in both serological tests, the highest frequency of positivity... (Complete abstract, access undermentioned eletronic address) / Mestre

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