Androgens are critical for the development and maintenance of adult male characteristics such as muscle mass and sexual function. Consequently, the established decline with age of serum testosterone (T) in males has major health implications. While the androgen receptor (AR) is the major mediator of genomic androgen action and is required for the development of the male phenotype, reproductive organs and the maintenance of male secondary sexual characteristics, it is the entrance of androgens into the cell that mediates the activation of the AR and the subsequent modulation of expression of androgen regulated genes. Testosterone, biologically the most important androgen in male serum, circulates either free, loosely bound to albumin or tightly bound to sex hormone binding globulin (SHBG). Each of these forms of serum T have different abilities to enter cells, and which proportion of serum T is capable of entering cells and initiating the androgen signalling cascade, thereby leading to the activation of the AR has not been precisely defined. The AR amino terminal domain (NTD) is responsible for the majority of the ability of the AR to activate genes but the relative roles of the two activation functions in the AR NTD (activation functions 1 and 5; AF1 and 5) have not been precisely defined while the role of the AF2 surface which forms in the ligand binding domain upon agonist binding is responsible for interactions with key coregulators and also with the NTD in the amino-carboxyl (N/C) interaction. Our laboratory has recently identified a region within AF5 between amino acids 500-535 to which somatic mutations in castrate resistant prostate tumour samples collocate. Due to the lack of functional information on the AF5 region and the NTD in general, the function of this region and the functional consequences of the mutations remain to be defined. The objectives of this thesis were to develop a specific mammalian cell based bioassay capable of reliable measuring T in serum and to determine the ability of this bioassay to measure a physiologically relevant fraction of T in serum. Additionally, this thesis aimed to determine the relative contributions and roles of the activation functions of the AR to overall AR transcriptional activity along with the functional consequences for AR signalling of prostate cancer mutations which have previously been identified in the AF5 region of the AR NTD. The mammalian-cell based bioassay developed in this thesis is capable of sensitively and reliably measuring serum T. However, evaluation of this bioassay utilising approximately 1000 serum samples from the Florey Adelaide Male Aging Study reveals that this bioassay measures a fraction of T in serum that most closely relates to serum T. Furthermore, this measure does not correlate more strongly with grip strength, sexual function or waist circumference than the existing immunoassay-based measures of serum T, highlighting the limitations of utilising a static mammalian cell-based androgen bioassay to measure physiological levels of serum T in males. The investigation of the roles of the activation functions in the AR in this thesis have revealed that while the AF1 domain is responsible for the majority of the transactivation activity of the AR, AF5 and AF2 govern the sensitivity and cellular response of the AR to androgens by providing protein and interdomain interaction interfaces. Furthermore, the evidence in this thesis demonstrates that the AR requires interdomain communication for sensitive AR signalling. Finally, the findings in this thesis demonstrate that the AF5 surface is required for the N/C interaction and coregulator interactions while advanced prostate cancer mutations identified within this region confer increased transactivation activity of the AR in the presence of high cellular levels of coregulators. Collectively, the findings in this thesis provide several novel insights into the mechanism of action of serum androgens and challenges several long held assumptions of androgenic action in males. These findings also delineate a mechanism of treatment failure in advanced prostate cancer, provide a novel model for the events leading to sensitive AR transactivation and contribute to the understanding of physiologically relevant levels of serum T. / Thesis (Ph.D.) -- University of Adelaide, School of Medicine, 2008

Phosphorus is the main limiting nutrient for plant growth in large areas of the world and the availability of phosphorus to plants and microbes can be strongly affected by soil properties. Even though the phosphorus cycle has been studied extensively, much remains unknown about the key processes governing phosphorus availability in different environments. In this thesis the complex dynamics of soil phosphorus and its availability were studied by relating various phosphorus fractions and soil characteristics to microbial respiration kinetics. The soils used represent a range of aluminium, iron, carbon and total phosphorus content, and were located in four different biomes: subtropical forest, warm temperate forest, boreal forest and tundra. The results showed that NaOH extractable phosphorus, a fraction previously considered to be available to plants only over long time scales, can be accessed by microbes in days or weeks. Microbial phosphorus availability was not related to aluminium or iron content in any of the studied systems, not even in highly weathered soils with high aluminium and iron content. This is in contrast with other studies of soils with high sorption capacity and shows the variability of factors that govern phosphorus availability in different environments. In the boreal forest chronosequence, no difference could be seen with age in total phosphorus content or concentrations of occluded phosphorus forms. However, there were lower concentrations of labile phosphorus forms in older systems, which were correlated with a decrease in microbial respiration. This was most likely related to organic matter quality in the system, and not to geochemical factors. Phosphorus availability was linked to differences in topography (water regime) and vegetation in the tundra ecosystems. The results suggest that the availability of phosphorus, both for microbes and plants, was lower on the meadow vegetation sites compared to the two types of heath vegetation. Many factors are important for phosphorus availability in soils, but these results suggest that microbes can access less available phosphorus if not restricted by carbon, and this may be important in regard to forest management practices as well as effects of environmental change.

published_or_final_version / Environmental Management / Master / Master of Science in Environmental Management

i ABSTRACT I investigated aspects of the management of subterranean termites in the field by evaluating the protective values of preservative treated softwoods and evaluating the potential of softwoods conditioned by decay fungi or heat to produce termite monitoring and aggregating devices. As part of the investigation, it was necessary to: · develop assay techniques suitable for research into the natural resistance of timbers, the efficacy of wood preservatives and termite biology and foraging behaviour; and quantify variation in termite responses to untreated softwoods in various situations in the field using these techniques. Field bioassays were conducted with two Australian Coptotermes species to determine protection of pine sapwood afforded by borate compounds (Chapter 2). Coptotermes lacteus (Froggatt) was examined in winter and C. acinaciformis (Froggatt) in winter, spring and summer at two localities in Queensland. Hoop-(Araucaria cunninghamii Ait ex D. Don) and slash-pine (Pinus elliottii Engelm.)-sapwood blocks were placed in plastic containers and attached to termite mounds or infested trees. Termite response was determined by mass losses over five weeks and modelled. Coptotermes lacteus was relatively benign in winter and for 1.0 and 2.5% mass losses of hoop-pine sapwood, borate retentions of 0.20 and 0.06% mass/mass (m/m) boric acid equivalent (BAE), respectively, were derived. With C. acinaciformis, for 1.0 and 5.0% mass losses of hoop-pine sapwood, borate retentions ranged from 0.49 and 0.25% BAE, respectively, in winter to 0.76 and 0.47% BAE, respectively, in late summer. For 1.0 and 5.0% mass losses of slash-pine sapwood in summer, borate retentions of 0.58 and 0.14% BAE, respectively, were derived for C. acinaciformis. These results support conclusions from earlier laboratory studies conducted elsewhere. Prevention of damage by field populations of termites was not achieved. Marked intercolony variation in response to borate compounds was demonstrated, highlighting the usefulness of this rapid technique. The need for long-term field exposure trials to calibrate the lunch-box technique and to facilitate technology transfer to industry, is indicated. For comparison, termite response to white cypress (Callitris glaucophylla Thompson and Johnson), a naturally termite resistant timber, is also reported. A field bioassay was conducted to evaluate responses of C. acinaciformis to untreated southern-yellow-pine (Pinus spp.), hoop-pine and slash-pine blocks (Chapter 3). Sapwood blocks were placed in plastic containers and attached to trees infested with the termites during summer. Containers with a test block of each of the timber species were prepared. These and other containers were exposed to termites for 10 weeks. Termite feeding response (“voracity”) was assessed for all blocks by determining the mass of wood consumed. Termite voracity differed among trees. Variation in termite responses to individual hoop-pine feeder-blocks within a container was appreciable and consistent between containers. Variation in termite responses to the test blocks was appreciable, with hoop pine being significantly more susceptible to termite attack than either slash pine or southern yellow pine. Southern yellow pine is a generic name for five species of pine which are difficult to separate reliably. Therefore, caution should be exercised when using “southern yellow pine” to ensure that samples are at least from the same species of pine and preferably from the same length of timber. The sensitivity of both the bioassay and the statistical tests was demonstrated. Of particular interest was the similar level of termite response to the test timbers in the two sections of the experiment. The usefulness of feeder-blocks to initiate reliable attack rather than to promote sustained attack, is worth further attention. Sapwood stakes of Australian-grown hoop pine, slash pine, radiata pine and North American-grown southern yellow pine were exposed to subterranean termite attack in an in-ground bioassay (Chapter 4). Stakes in bait containers and bare stakes were attacked by C. acinaciformis and Schedorhinotermes intermedius (Brauer). Susceptibility of these timbers was evaluated with regard to potential for termite monitoring and aggregation. Variation between timbers and difference between termite species are described. The relevance of these data to suppressing foraging populations of subterranean termites, in Australia, using insect growth regulators, is discussed. Four field bioassays were conducted using two species of Australian subterranean termites, C acinaciformis and C. lacteus (Chapter 5). Hoop-pine blocks decayed using 14 fungi for four weeks and for 10 weeks were subsequently conditioned using dry heating, freezing or moist heating, and then exposed to termites. Blocks decayed by Gloeophyllum abietinum [Persoon: Fries] Murrill, Trametes lilacino-gilva (Berkeley) Wright & Deschamps, Schizophyllum commune Fries and Rigidoporus undatus (Persoon) Donk and subsequently oven dried, were attacked the most. Oven drying generally enhanced termite feeding responses in these two field bioassays. In a third bioassay, blocks of five softwood timber species conditioned with two high temperature regimes were exposed to C. acinaciformis. Slash pine was attacked the least. Mean mass losses for hoop pine, Caribbean pine (Pinus caribaea Morelet), radiata pine and the interspecific F1 hybrid between Caribbean and slash pine were not significantly different. For C. lacteus, slash and hoop-pine blocks were attacked the least while radiata pine was attacked the most. Additional heating at 105°C increased the susceptibility of most timber species. Relationships between heating duration and the consumption of hoop-pine-sapwood blocks were determined using results from a fourth bioassay. Heating the timber increased feeding responses for both termite species. The selective and rapid response of the termites highlights the usefulness of this field technique for multiple-choice bioassays. The cyclodiene termiticides were withdrawn from use, in Australia, as from 30 June 1995. Implications for the management of subterranean termites in Queensland are discussed. Community perceptions of the use of chemical pesticides and cost do not favour the widespread use of preservative treated timber in buildings. An integrated pest management approach to termite control is the likely direction for the future in Australia. A range of building techniques, regular inspections, and attention to factors that predispose existing buildings to termite attack can reduce the risk of damage from termites, but do not eliminate that risk. There will be an increased need for reliable preventative and remedial treatments involving bait technology. Much more work is needed on termite biology, foraging behaviour and bait toxicants before a baiting system can be confidently commercialised in Australia. Directions for research in Queensland are discussed.

Thesis (M.S.)--University of North Carolina at Wilmington, 2005. / Includes bibliographical references (leaves: 48-49)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Baccharis dracunculifolia is a medicinal species, popularly known as broom, and has its use based on the preparation of infusions of leaves for the treatment of gastric disorders, physical fatigue, lack of appetite, fever diseases and organic weakness. Its essential oil stands out in the perfumery industry as well as in biological studies. The study aimed to evaluate the potential antiproliferative, genotoxic and antigenotoxic of two populations of B. dracunculifolia on the test systems Allium cepa L. and Lactuca sativa L. as well as performs the phytochemical analysis of the aqueous extract and oil obtained from the leaves. The material used in the preparation of infusions and oil consisted of two populations collected in Santa Maria (SM) and São Pedro do Sul (SPS), which part was dehydrated to prepare infusions in 5g.L-1 and 10g.L -1 concentrations, and the other part was frozen for later extraction of essential oil. To determine the antiproliferative, genotoxic and antigenotoxic activities, the effects of aqueous extracts and oil on the A. cepa test and L. sativa systems. The experiment whith A. cepa consisted of twelve groups of four bulbs, each group corresponding to a treatment. The bulbs were rooted in water an than subjected to treatments for 24 hours. The experiment whith L. sativa consisted of nine groups of four Petri dishes. Each plate was soacked whith about 7 mL of extract, which were in contact with the seeds for 72 hours. After this period the roots of A. cepa and L. sativa seeds were collected, fixed in Carnoy 3:1 and stored in refrigerated in 70% ethanol until the preparation of the slides. The determination of phenolic compounds from aqueous extracts was performed by the High Performance Liquid Chromatography (HPLC). The compounds of the essential oil were determined by Gas Chromatography (GC). The genotoxicity data were subjected to analysis Chi-square test, whereas the data of the chromatographic analysis were subjected to Tukey test (p <0.05). The results of the A. cepa test showed that extracts from B. dracunculifolia have antiproliferative and antigenotoxic potential as well as the essential oil of the SPS population, while the oil SM population induces cell proliferation. For L. sativa test, aqueous extracts of the two populations demonstrated atiproliferativo effect, except the aqueous extract of SPS population at a concentration of 10 g.L-1, in which there was an increase in cell proliferation. Chromatographic analysis indicated as major compounds, p-coumaric acid to the aqueous extracts and E-nerolidol to the essential oil. / Baccharis dracunculifolia DC. é uma espécie medicinal, conhecida popularmente como vassourinha, e tem sua utilização baseada no preparo de infusões das folhas para o tratamento de distúrbios gástricos, cansaço físico, inapetência, afecções febris e debilidade orgânica. Seu óleo essencial se destaca na indústria de perfumaria, bem como em estudos biológicos. O trabalho teve por objetivos avaliar o potencial antiproliferativo, genotóxico e antigenotóxico de duas populações de B. dracunculifolia sobre os sistemas teste de Allium cepa L. e Lactuca sativa L., bem como realizar a análise fitoquímica do extrato aquoso e do óleo obtidos a partir das folhas. O material utilizado no preparo das infusões e do óleo constou de duas populações coletadas em Santa Maria (SM) e São Pedro do Sul (SPS), do qual parte foi desidratado para preparo das infusões nas concentrações 5g.L-1 e 10g.L-1 e outra parte foi congelada para posterior extração do óleo essencial. Para determinação das atividades antiproliferativa, genotóxica e antigenotóxica, foi avaliado o efeito dos extratos aquosos e do óleo sobre os sistemas teste de A. cepa e L. sativa. O experimento com A. cepa constou de doze grupos de quatro bulbos, cada grupo correspondendo a um tratamento. Os bulbos foram enraizados em água e posteriormente submetidos aos tratamentos por 24 horas. O experimento com L. sativa constou de nove grupos de quatro placas de Petri, cada grupo correspondendo a um tratamento, onde foram dispostas 50 sementes por placa. Cada placa foi embebida com aproximadamente 7 mL de extrato, os quais ficaram em contato com as sementes por 72 horas. Após esse período as raízes de A. cepa e as sementes de L. sativa foram coletadas, fixadas em Carnoy 3:1 e armazenadas sob refrigeração em etanol 70% até o preparo das lâminas. A determinação dos compostos fenólicos presentes nos extratos aquosos foi realizada pela técnica de Cromatografia Líquida de Alta Eficiência (CLAE). Os compostos do óleo essencial foram determinados pela técnica de Cromatografia Gasosa (CG). Os dados obtidos da análise de genotoxicidade foram submetidos ao teste Qui-quadrado, enquanto que, os dados da análise cromatográfica foram submetidos ao teste de Tukey (p<0,05). Os resultados do teste de A. cepa demonstraram que os extratos de B. dracunculifolia possuem potencial antipoliferativo e antigenotóxico, bem como o óleo essencial da população de SPS, enquanto que o óleo da população SM induziu a proliferação celular. Para o teste de L. sativa, os extratos aquosos das duas populações demonstraram efeito antiproliferativo, com exceção do extrato aquoso da população SPS na concentração de 10 g.L-1, no qual houve um aumento da proliferação celular. As análises cromatográficas indicaram como compostos majoritários, o ácido p-cumárico para os extratos aquosos e o E-nerolidol para o óleo essencial.

Made available in DSpace on 2014-06-11T19:26:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-07-13Bitstream added on 2014-06-13T19:12:59Z : No. of bitstreams: 1 rossi_jr_me_jabo.pdf: 1016425 bytes, checksum: 885b9804e9660ef2c359c1d1eb71a757 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Dentre os microrganismos empregados no controle de mosquitos, a bactéria Bacillus thuringiensis var. israelensis (Bti) se destaca por apresentar atividade tóxica contra insetos da ordem Diptera, na qual produz inclusões cristalinas compostas pelas proteínas Cry, que são codificadas por genes cry presentes em um único plasmídeo. A produção dessas proteínas em grande escala está relacionada a mecanismos transcricionais, como por exemplo, a expressão de um gene sob o controle de um promotor forte. Tendo em vista que a bactéria Bti SPS1 (Patente PI0200228-0) foi isolada do território brasileiro e que possui potencial para o controle de vetores por produzir uma maior quantidade de esporos/cristais em menor tempo, este trabalho teve por finalidade a caracterização da região promotora dos genes cry4Aa, cry4Ba e cry11Aa de Bti SPS1 pelas técnicas de PCR e “Southern blotting”. Em associação a estas técnicas, foi realizado bioensaio para a verificação da mortalidade de larvas de Aedes aegypti. Os resultados das análises moleculares indicaram homologia no perfil de hibridização da região promotora da linhagem padrão Bti T14-001 e do isolado Bti SPS1. A quantificação das suspensões em espectrofotômetro (DO600nm) e a leitura de esporos em câmara de Neubauer, revelaram que o isolado Bti SPS1 produz uma maior quantidade de esporos/cristais em relação à linhagem padrão Bti T14-001. O bioensaio apresentou elevados índices de mortalidade. Estes resultados tornam a bactéria Bti SPS1 uma fonte promissora para novas formulações visando o controle de vetores. / Among the microorganisms used for mosquitoes’ control the bacterium Bacillus thuringiensis var. israelensis (Bti) is frequently considered since it presents toxic activities’ against Diptera, producing crystal inclusions including Cry proteins, which are coded by a sole plasmid borne set of genes. The production of these proteins in large scale is related to transcriptional mechanisms, as an example, a particular gene expression controlled by a strong promoter. Since the Bti bacterial isolate SPS1 (Patent PI0200228-0) was isolated within the Brazilian territory and exhibits potential for the control of vector insects due to a higher crystal production ability in shorter time period, this work had as objective the characterization of the promotion region for the genes cry4Aa, cry4Ba e cry11Aa using PCR and “Southern blotting” techniques. Also some bioassays using Aedes aegypti larvae were carried out. The results from the molecular analysis have indicated homology for the hybridization profile from the promoter region from the type strain of Bti T14-001 and that of the SPS1 isolate. Spectrofotometric (OD600nm) and Neubauer chamber measures have revealed that the SPS1 isolate produces a higher amount of spore/crystal as compared to the Bti T14-001 strain. The bioassay presented higher mortality levels. These results seem to indicate that the isolate SPS1 is a promising bacterial strain to be used on formulations able to control insect vector pests.

Made available in DSpace on 2014-10-09T12:42:17Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:00Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Made available in DSpace on 2014-10-09T12:41:24Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:03:39Z (GMT). No. of bitstreams: 0 / Dissertação (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Made available in DSpace on 2014-10-09T12:42:03Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:04:44Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP