The mammalian spinal cord contains the neuronal circuitry necessary to generate rhythmic locomotor activity in the absence of inputs from the higher brain centre or sensory system. This circuitry is regulated by local neuromodulatory inputs, which can adjust the strength and timing of locomotor output. The free radical gas nitric oxide has been shown to act as an important neuromodulator of spinal circuits, which control locomotion in other vertebrate models such as the tadpole and lamprey. Despite this, the involvement of the NO-mediated soluble guanylate cyclase/cyclic guanosine monophosphate secondary messenger-signalling pathway (NO/sGC/cGMP) in mammalian locomotion has largely been under-investigated. The NADPH diaphorase histochemical reaction was used to identify sources of NO in the lumbar spinal cord. The largest population NADPH diaphorase reactive neurons were located in the dorsal horn, followed by the laminae of the ventral horn, particularly around the central canal (lamina X) and lamina VII. NADPH diaphorase reactive neurons were found along a rostrocaudal gradient between lumbar segments L1 to L5. These results show that that discrete neuronal sources of NO are present in the developing mouse spinal cord, and that these cells increase in number during the developmental period postnatal day P1 – P12. NADPH diaphorase was subsequently used to identify NADPH diaphorase reactive neurons at P12 in the mouse model of ALS using the SODG93A transgenic mouse. Physiological recordings of ventral root output were made to assess the contribution of NO to the regulation induced rhythmic fictive locomotion in the in vitro isolated spinal cord preparation. Exogenous NO inhibits central pattern generator (CPG) output while facilitating and inhibiting motor neuron output at low and high concentrations respectively. Removal of endogenous NO increases CPG output while decreasing motor neuron output and these effects are mediated by cGMP. These data suggest that an endogenous tone of NO is involved in the regulation of fictive locomotion and that this involves the NO/sGC/cGMP pathway. Intracellular recordings from presumed motor neurons and a heterogeneous, unidentified sample of interneurons shows that NO modulates the intrinsic properties of spinal neurons. These data suggest that the net effect of NO appears to be a reduction in motor neuron excitability.

Parmi les différents minéraux argileux, la sépiolite, qui est un silicate fibreux naturel, est un potentiel nano-transporteur prometteur pour le transfert non-viral de biomolécules. Il a en effet été montré que la sépiolite interagissait avec des molécules biologiques telles que les lipides, les polysaccharides et les protéines. Dans ce travail, nous démontrons que la sépiolite interagit également efficacement avec différents types de molécules d'ADN (génomique, plasmidique, oligonucléotides simple et double brin), et nous présentons la première étude détaillée sur les mécanismes d'interaction entre la sépiolite et l'ADN, ainsi qu’une caractérisation physico-chimique de bionanocomposites ADN-sepiolite. Une analyse spectroscopique a montré tout d’abord que l’interaction de l'ADN avec la sépiolite était plus forte en présence de polycations, la valence de ces derniers accroissant le rendement d’absorption, et deuxièmement, que l'ADN ainsi adsorbé pouvait être récupéré avec un rendement modulé par la présence d’EDTA, la structure de l'ADN et son activité biologique étant conservées. Par spectroscopie infrarouge à transformée de Fourier (FTIR) nous avons identifié les groupes silanol externes comme les principaux sites d'interaction avec l'ADN. Nous avons ensuite prouvé qu'il est possible d'utiliser la sépiolite pour extraire l'ADN de bactéries, pour la purification de l'ADN et pour la purification de toute contamination bactérienne. En combinant la microscopie à fluorescence, la microscopie électronique à transmission (MET), la vidéo-microscopie et l’analyse par cytométrie en flux (FACS), nous avons montré que la sépiolite peut être spontanément internalisée dans des cellules de mammifère par le biais de deux voies, l’endocytose et la macropinocytose. En tant que preuve de concept, nous montrons que la sépiolite est capable de transférer de manière stable l'ADN de plasmide dans des bactéries et des cellules de mammifères. Il a également été prouvé qu’en incubant des bactéries avec des bionanocomposites ADN-sepiolite, initialement préparés en présence d'une faible concentration en cations divalents et avec de la sépiolite traitée aux ultrasons (sSep), il était possible d'augmenter l'efficacité de la transformation bactérienne 20 à 30 fois par rapport aux méthodes basées sur l'«effet Yoshida». En outre, nous montrons que l'efficacité du transfert de gènes par la sépiolite peut être optimisée : l'utilisation de sSep et l'exposition à la chloroquine augmentent d’un facteur 100 et 2, respectivement, l’efficacité de transfection. Ces résultats ouvrent la voie à l'utilisation de bionanocomposites à base de sépiolite comme de nouveaux potentiels nano-transporteurs hybrides potentiels, à la fois pour la thérapie génique et le développement de nouveaux modèles biologiques en sciences fondamentales et appliquées. / Among the various clay minerals, sepiolite, which is a natural fibrous silicate, isa potential promising nanocarrier for the non-viral transfer of bio-molecules. Indeed,sepiolite has been shown to interact with biological molecules such as lipids,polysaccharides and proteins. Here, we show that sepiolite efficiently binds differenttypes of DNA molecules (genomic, plasmid, single strand and double strandoligonucleotides), introducing the first detailed study on the interaction mechanismsbetween sepiolite and DNA, as well as the physicochemical characterization of theresulting DNA-sepiolite bionanocomposites. The interaction mechanisms aresuggested to be electrostatic interactions, van der Waals forces, cation bridges, andhydrogen bonding. Spectroscopy analysis showed that the binding of DNA to sepiolitewas increased by polycations with valence dependent efficiency, and the DNApreviously adsorbed could be recovered with an efficiency that could be modulatedusing a chelating agent (EDTA), preserving the DNA structure and biological activity.Fourier-transform infrared spectroscopy identified the external silanol groups as themain sites of interaction with the DNA. It was proved that it is possible to use sepiolitefor extracting DNA from bacteria, for DNA purification and for purification from bacterialcontamination. By combining fluorescence microscopy, transmission electronmicroscopy (TEM), time-lapse video microscopy and flow cytometry analysis (FACS),we show that sepiolite can be spontaneously internalized into mammalian cells throughboth endocytic and non-endocytic pathways. As a proof of concept, we show thatsepiolite is able to stably transfer plasmid DNA into bacteria and mammalian cells. Itwas also proved that with the incubation of bacteria with the Sep/DNAbionanocomposite initially prepared in the presence of a low concentration of divalentcation, and using sonicated sepiolite (sSep), it is possible to increase the bacterialtransformation efficiency from 20 to 30-fold compared to previously reported methodswhich are based in the “Yoshida effect”. Additionally, we show that the efficiency ofsepiolite-mediated gene transfer can be optimized: the use of sSep and the exposureto the endosome disrupter chloroquine 100-fold and 2-fold stimulated DNA transfectionefficiency, respectively. These results open the way to the use of sepiolite-basedbionanocomposites as a novel class of hybrid nanocarriers for both potential genetherapy and the development of novel biological models of interest for academic andapplied sciences.

Células de mamíferos são os hospedeiros mais frequentemente utilizados para a fabricação de proteínas biofarmacêuticas e para a produção de vacinas virais, A qualidade é um elemento-chave para o estabelecimento de um processo de bioconversão eficiente. No presente trabalho utilizamos a linhagem de células BHK- 21C13(Baby Hamster Kidney) adaptadas para cultivo em suspensão. O uso de Soro Fetal Bovino (SFB) é tradicionalmente utilizado, sendo considerado um suplemento universal, pois permite o crescimento em várias linhagens de células de mamíferos; porém, uso de SFB apresenta risco de infecção por prions, variabilidade entre lotes e aumento no custo em etapa de purificação (Downstream processing). O objetivo do presente trabalho foi comparar o cultivo de células BHK-21 C13 entre dois meios suplementados com SFB e sem SFB, através do estudo cinético para cultivo em suspensão estático e agitado com frascoT, frasco spinner e biorreator, respectivamente. Os parâmetros; Xmáx e µmáx, não foram significativamente influenciados pelo meio de cultura em cultivo estático, em cultivo com agitação em frasco spinner e também no cultivo em biorreator. O tempo de duplicação ficou próximo para todas as condições testadas. A produtividade alcançada foi: 0,032x106 cel/mL.h-1 para o meio com SFB e 0,031 X106 cel/mL.h-1 para o meio sem SFB. Ao final do processo foi possível obter uma concentração celular em torno de 4,7x106 cel/mL, tanto para o cultivo com SFB quanto para o cultivo sem SFB. Dessa forma, o uso de meio de cultivo sem SFB não alterou os principais parâmetros cinéticos, não apresentando as desvantagens do uso do SFB. / Mammalian cells are the most frequently used hosts for the production of biopharmaceutical proteins and viral vaccines. Quality is a key element for the establishment of an efficient bioconversion process. In this work, we used the cell line Baby Hamster Kidney C13 (BHK-21 C13) adapted to suspension culture was used. Fetal Bovine Serum (FBS) is traditionally used and it is considered a universal insert due to its power to increase cell growth in this kind of animal cells. However, the utilization of FBS introduces risks of infection from prions, variability between batches and increase in cost associated to purification stages (downstream processing). This work aimed to compare the kinetic behaviors of BHK-21 C13 cells in two media supplemented with FBS and without FBS using both one static and two suspension systems, T-flask, spinner flask and bioreactor respectively. The parameters; Xmax and µmax were not significantly influenced by the culture medium in T- flask culture static, in spinner flask cultivation and were neither significantly influenced by growing in culture media stirred bioreactor. The doubling time was close to all conditions tested. At the end of the growth phase it was possible to obtain a nearby cell concentration of 4.7 x 106 cells / ml, both for cultivation with FBS as for FBS without cultivation. Thus, the use of culture medium without FBS did not affect the main kinetic parameters. Besides, it does not show the disadvantages of culture media using FBS.

Atualmente, o Brasil encontra-se na privilegiada posição de maior produtor e exportador mundial de carne bovina, tornando a pecuária uma das atividades nacionais mais importantes e rentáveis. Este dado enfatiza a importância de pesquisa e desenvolvimento em reprodução bovina, especialmente em hormônios estimuladores da ovulação, tais como a gonadotrofina coriônica equina (eCG). Os produtos comerciais à base de eCG comercialmente disponíveis são purificados a partir do sangue de éguas gestantes, apresentando variabilidade de lote para lote e presença de contaminantes. Estes fatos, juntamente com a limitação do material de partida (sangue equino), enfatizam a necessidade de haver um sistema de expressão de eCG recombinante passível de ser explorado comercialmente. Neste quesito, as células de mamíferos se mostram um sistema robusto para tal finalidade, visto que são capazes de adicionar modificações pós-traducionais às cadeias polipeptídicas, tais como a glicosilação, o que é essencial para o correto dobramento, maturação e montagem das duas subunidades, além de interferir diretamente com a meia-vida, o reconhecimento do receptor, a solubilidade e a atividade biológica das proteínas. No entanto, mesmo entre os sistemas de expressão heteróloga em células de mamífero, encontra-se muita variabilidade nos padrões de glicosilação adicionado. No presente trabalho, foi realizado um estudo comparativo através da clonagem e expressão de uma forma fusionada de eCG (reCGβα) em duas linhagens celulares diferentes: (1) CHO-DG44, um dos sistemas de expressão mais utilizados pelas indústrias farmacêuticas, capaz de adicionar N-glicanos complexos; e (2) 293T, uma linhagem humana capaz de produzir glicoproteínas carreando oligossacarídeos complexos e sialilados. Os resultados de atividade biológica (in vitro e in vivo) apontam uma maior atividade de reCG produzido por células CHO-DG44. O perfil de N-glicosilação de reCG produzido pelas células CHOD-G44 assemelhou-se mais à eCG selvagem, quando comparado a reCG produzido por células 293T. Por fim, estudos clínicos foram realizados com reCG produzido em meio livre de soro fetal bovino e parcialmente purificado, onde atividade específica de reCG produzido por células CHO-DG44 mostrou-se similar ao produto comercial selvagem. / Brazil is currently the major beef producer and exporter, rendering to livestock one of the country´s most economically relevant activities. This emphasizes the importance of research and development in bovine reproduction, especially at ovulation-stimulatory hormones, such as equine gonadotropin (eCG). The commercially available eCG-based products are purified from blood of pregnant heifers, presenting batch-to-batch variability and the presence of contaminants. These facts, together with the limitation of the bulk material (equine blood), emphasize the need of an eCG expression system able to be commercially explored. In this aspect, mammalian cells are a robust system, capable of add post-translational modifications to polypeptide chains, such as glycosylation, which is essential for the correct folding, maturation and assembly of both eCG subunits. In addition, glycosylation directly interferes with the protein half-life, receptor recognition, solubility and biological activity. In the present work, a comparative study was carried out by cloning and expressing a fusion form of eCG (reCGβα) in two different mammalian cell lines: (1) CHO-DG44, one of the most used by pharmaceutical companies expression systems, capable of add complex-type N-glycans; and (2) 293T, a human cell line capable of produce glycoproteins carrying complex and sialylated oligosaccharides. The in vitro and in vivo biological activity results show a higher potency of reCG produced by CHO-DG44 cells. The N-glycosylation pattern produced by CHO-DG44 cells was more similar to native eCG in comparison to the N-glycosylation produced by 293T cells. Finally, clinical studies were performed with serum absent media produced and partially purified reCG, showing that the specific activity of reCG produced by CHO cells was similar to the commercial wild type product.

O objetivo deste trabalho foi estudar o processo pelo qual hormônios e fatores de crescimento controlam a proliferação celular em mamíferos. O modelo experimental utilizado foi linhagens de células estabelecidas em cultura. Os estudos centraram-se em dois tipos básicos de células: fibroblastos e células adrenais e o ataque experimental foi feito sob dois pontos de vista: bioquímico e genético. O ataque bioquímico envolveu desenvolver estudos cinéticos da síntese de DNA não só durante o carenciamento de células para soro, como também durante a reestimulação de células carenciadas por:soro, hormônios e fatores de crescimento. Medidas do conteúdo intracelular de cAMP foram efetuadas com o intuito de adquirir informações à respeito do mecanismo de ação destes fatores. Um modelo de ciclo celular foi proposto no qual o controle do crescimento seria exercido através de reguladores positivos e negativos que agiriam estimulando ou inibindo a passagem de células do estado de repouso (Go) para a fase proliferativa. Entre os reguladores positivos (estimuladores) do sistema fibroblasto, encontra-se hormônios clássicos, como esteróides e insulina, e fatores de crescimento de natureza hormonal como EGF, PF (fator proteico extraído de glândulas pituitárias) e prostaglandina F2α. O esteróide hidrocortisona pode agir como regulador negativo, inibindo o crescimento de fibroblastos. Medidas do período de tempo transcorrido desde a estimulação de células carenciadas (Go), até o aparecimento da onda de síntese de DNA (período definido operacionalmente como Gl) foram feitas. Em fibroblastos 3T3 este período foi de 12 a 13 horas tanto para células estimuladas com soro como com hormônios clássicos (hidrocortisona, insulina) ou fatores de crescimento (EGF, PF) ou ainda com combinações deles (EGF + PF + insulina; PF + hidrocortisona; PF + hidrocortisona + insulina). No sistema células adrenais, adrenocorticotropina (ACTH) foi o único hormônio clássico que apresentou atividade sobre o crescimento destas células e também o único efetuador negativo encontrado. Neste sistema PF mostrou-se como o único fator com atividade estimulatória sobre o crescimento. Gl aqui foi de 11 horas tanto para células estimuladas com soro como com PF. Além disso os hormônios clássicos hidrocortisona e insulina não apresentaram atividade estimulatória por si só ou em combinação com PF. A análise da ação de hidrocortisona no sistema fibroblasto e de ACTH no sistema células adrenais estimuladas, forneceu evidências de que após deixar Go, em direção a S, numa certa altura de Gl as células tornam-se irreversivelmente comprometidas com o processo replicativo. Este comprometimento parece ocorrer 5 horas antes de S, sendo referido como Glc. Em face destes resultados foi proposto que os reguladores agem estimulando ou inibindo a transição Go → Glc. Na tentativa de obter maior definição do sistema de controle do crescimento, aproveitamo-nos das vantagens oferecidas pelo modelo experimental usado, para a busca de mutantes do tipo regulatório. Esta busca resultou no isolamento das linhagens ST1 e AR-1, derivadas, respectivamente, de fibroblastos 3T3 e células adrenais Y-l. Entre os vários aspectos interessantes da linhagem ST1 destaca-se: a) o dramático efeito de hidrocortisona causando mudança nas características das células as quais passam de um fenótipo tipicamente transformado para normal. Este fenômeno foi observado tanto \"in vitro\" (através de medidas de parâmetros de crescimento) como \"in vivo\" (através de ensaios de tumorogenicidade); b) as alterações morfológicas de caráter antagônico provocadas, por um lado, pela adição de hidrocortisona (causando achatamento) e, por outro, pela retirada do soro ou adição de cAMP ao meio de cultura (arredondamento). Através do estudo da ação de inibidores, obteve-se evidências do envolvimento de microtúbulos nestas alterações morfológicas. A análise do conteúdo intracelular de cAMP indicou que este nucleotídeo não atua como mediador da ação de hidrocortisona. Sua ação parece ser devida à indução de alterações no sistema superfície celular - membrana -citoesqueleto. Ao contrário de outros variantes de células Y-l resistentes à ACTH, células AR-1 mostraram-se também resistentes a cAMP. A utilidade destas células nos estudos da postulada mediação deste nucleotídeo na ação de ACTH, é óbvia. / The aim of this work was to study the process by which hormones and growth factors control proliferation of mammalian cells. Cell lines established in culture were used as the experimental model. The studies were centered on two basic types of cells: fibroblasts and adrenal cells and the experimental approach was made from two viewpoints: biochemical and genetic. The biochemical approach involved kinetic studies of the DNA synthesis process not only during serum starvation but also during restimulation of serum starved cells by serum, hormones and growth factors. Intracellular cyclic AMP determinations were made in order to gain informations on the mechanism of action of these factors. A cell cycle model was proposed in which cell growth control would be exerted by positive and negative regulators that would act by stimulating or inhibiting the flow of cells from a resting state (Go) to the proliferative phase. Among the positive regulators (stimulators) found for the fibroblast system are: classical hormones, like steroids and insulin, and growth factors of homonal nature, like EGF, PF (protein factor extracted from pituitary glands) and prostaglandin F2α. The steroid hydrocortisone can also act as a negative regulator, inhibiting fibroblast growth. Measurements of the time interval between stimulation of serum starved (Go) cells and the onset of DNA synthesis (period that is operationally defined as Gl) were made. In 3T3 fibroblasts this period was 12 to 13 hours for cells stimulated not only by serum but also by classical hormones (hydrocortisone, insulin) or growth factors (EGF, PF) or even by combinations of these factors (EGF + PF + insulin; PF + hydrocortisone; PF + hydrocortisone + insulin). In the adrenal system, adrenocorticotropin (ACTH) was the only classical hormone to present activity on the growth of these cells and also the only negative regulator found. In this system PF was shown to be the only factor with growth stimulatory activity. GL was estimated as 11 hours for cells stimulated with serum or PF. Moreover hydrocortisone and insulin had no stimulatory activity \"per si\" or in combination with PF. The analysis of hydrocortisone action on the fibroblast system on one hand and of that of ACTH on the adrenal system, on the other, indicated that upon leaving Go, towards S, at a certain point in Gl, cells become irreversibly committed to the replicative process. This commitment seems to occur 5 hours before S and is referred to as Glc. In view of these data we proposed that regulators act by stimulating or inhibiting the transition Go → Glc. In an attempt to obtain a better definition of the growth control system and taking advantage of the experimental model utilized, we searched for mutants of the regulatory type. This search resulted in the isolation of the lines ST1 and AR-l from 3T3 fibroblasts and Y-l adrenal cells, respectively. Among several interesting aspects of the STl cell line, we point out: a) the dramatic effect of hydrocortisone changing the characteristics of these cells from a typically transformed phenotype to a normal pattern. This phenomenon was observed both \"in vitro\" (by measuring a number of growth parameters) and \"in vivo\" (by tumorogenicity assays). b) morphological alterations of antagonistic nature caused by hydrocortisone (flattening) on one hand, and by the removal of serum or cAMP addition to the culture medium (rounding) on the other. Evidence for the involvement of microtubules in these alterations were obtained through studies on the action of several inhibitors. Quantitative analysis of intracellular cAMP indicated that this nucleotide does not act as a mediator of hydrocortisone action. Rather, this action seems to be due to the induction of alterations on the cell surface-membrane-cytoskeleton system. Contrary to other variants of the Y-1 line which are resistant to ACTH, AR-1 cells are also resistant to cAMP. The usefulness of these cells in studies of the postulated mediation by cAMP of the ACTH action, is obvious.

Células de mamíferos são os hospedeiros mais frequentemente utilizados para a fabricação de proteínas biofarmacêuticas e para a produção de vacinas virais, A qualidade é um elemento-chave para o estabelecimento de um processo de bioconversão eficiente. No presente trabalho utilizamos a linhagem de células BHK- 21C13(Baby Hamster Kidney) adaptadas para cultivo em suspensão. O uso de Soro Fetal Bovino (SFB) é tradicionalmente utilizado, sendo considerado um suplemento universal, pois permite o crescimento em várias linhagens de células de mamíferos; porém, uso de SFB apresenta risco de infecção por prions, variabilidade entre lotes e aumento no custo em etapa de purificação (Downstream processing). O objetivo do presente trabalho foi comparar o cultivo de células BHK-21 C13 entre dois meios suplementados com SFB e sem SFB, através do estudo cinético para cultivo em suspensão estático e agitado com frascoT, frasco spinner e biorreator, respectivamente. Os parâmetros; Xmáx e µmáx, não foram significativamente influenciados pelo meio de cultura em cultivo estático, em cultivo com agitação em frasco spinner e também no cultivo em biorreator. O tempo de duplicação ficou próximo para todas as condições testadas. A produtividade alcançada foi: 0,032x106 cel/mL.h-1 para o meio com SFB e 0,031 X106 cel/mL.h-1 para o meio sem SFB. Ao final do processo foi possível obter uma concentração celular em torno de 4,7x106 cel/mL, tanto para o cultivo com SFB quanto para o cultivo sem SFB. Dessa forma, o uso de meio de cultivo sem SFB não alterou os principais parâmetros cinéticos, não apresentando as desvantagens do uso do SFB. / Mammalian cells are the most frequently used hosts for the production of biopharmaceutical proteins and viral vaccines. Quality is a key element for the establishment of an efficient bioconversion process. In this work, we used the cell line Baby Hamster Kidney C13 (BHK-21 C13) adapted to suspension culture was used. Fetal Bovine Serum (FBS) is traditionally used and it is considered a universal insert due to its power to increase cell growth in this kind of animal cells. However, the utilization of FBS introduces risks of infection from prions, variability between batches and increase in cost associated to purification stages (downstream processing). This work aimed to compare the kinetic behaviors of BHK-21 C13 cells in two media supplemented with FBS and without FBS using both one static and two suspension systems, T-flask, spinner flask and bioreactor respectively. The parameters; Xmax and µmax were not significantly influenced by the culture medium in T- flask culture static, in spinner flask cultivation and were neither significantly influenced by growing in culture media stirred bioreactor. The doubling time was close to all conditions tested. At the end of the growth phase it was possible to obtain a nearby cell concentration of 4.7 x 106 cells / ml, both for cultivation with FBS as for FBS without cultivation. Thus, the use of culture medium without FBS did not affect the main kinetic parameters. Besides, it does not show the disadvantages of culture media using FBS.

A population of spotted-tailed quolls Dasyurus maculatus was studied for three years (2002-2004) in the lower catchment of the Jacobs River, in the Byadbo Wilderness Area of southern Kosciuszko National Park, south-eastern New South Wales, Australia. Survey and monitoring of quoll latrine sites and prey populations, dietary analysis and live-trapping was carried out for one year before and two years after the widespread wildfires of January 2003, which had a very high impact on the study area. Survey for spotted-tailed quoll latrine sites was successful in locating a total of 90 latrine sites in the Jacobs River study area over the three years of the study. These were found throughout all parts of the topography among large, complex granite outcrops and along rocky sections of riparian habitat. After the fire in 2003, lower numbers of latrines were in use than observed pre-fire, and there was a lower level of usage (number of scats) of individual latrines. Continued monitoring in 2004 revealed that many latrines that had become inactive in 2003 following the fire were re-activated in the second breeding season following fire. 1466 spotted-tailed quoll scats were collected from latrines and live-trapped quolls over the three years of the study. Hair analysis from scats identified twenty-two different species of mammal in the diet of the spotted-tailed quoll from the Jacobs River study area, representing the majority of all prey identified (98.5% occurrence) and contributing almost all of the biomass consumed (99.6%). Medium-sized mammals were the most important prey category, followed by small mammals, large mammals (most likely taken as carrion) and non-mammalian prey (birds, reptiles, insects and plants). Brushtail possums were the most important single prey item by both frequency of occurrence and percentage biomass in all years, followed by lagomorphs (rabbits and hares), Rattus spp., and swamp wallabies. There was a significant difference in the composition of the diet by major prey category across the years of the study as a result of the fire, indicated by a shift in utilisation of food resources by quolls in response to significant changes in prey availability. Monitoring of prey populations revealed that brushtail possums, lagomorphs and bandicoots were all significantly less abundant in the study area in the winter directly following the fire, followed by a significant increase in abundance of lagomorphs, but not of possums, in the second winter after the fire. Quolls adapted well to this altered prey availability. While there was a significant decrease in occurrence of brushtail possum in scats after the fire, significantly more scats contained hair of lagomorphs, to the point where almost equal proportions of lagomorphs and possum hair occurred in scats by the winter of 2004. Other fire-induced changes to the diet were evident, such as a significant drop in the occurrence of small mammals in scats for both winters after the fire, and a peak in occurrence of large mammals in the winter directly following the fire that strongly suggests there was a short-term increase in the availability of carrion. A large, high-density population of spotted-tailed quolls was live-trapped and marked during the winter breeding season of 2002. Twenty-two quolls (13 male and 9 female) were present in the study area in 2002, and subsequent trapping over the 2003 and 2004 winter breeding seasons following the fire revealed that the high-intensity wildfire did not result in the extinction of the local population. There was evidence of a small, short-term decline in the number of quolls present in the study area in the 2003 breeding season, with 16 individual quolls captured. Males were outnumbered two-to-one by females, due either to mortality or emigration. Trapping in 2004 showed a recovery of the population to numbers exceeding that observed prior to the fire, with 26 individuals captured (16 male, 10 female), most likely as a result of immigration. There was some evidence that recruitment of young from the post-fire breeding season in 2003 was reduced because of the fire. This study took advantage of an unplanned wildfire event to monitor the response of a population of spotted-tailed quolls and their prey. In this regard it was fortuitous since it has been recognised that the use of replicates and controls in the study of the impacts of wildfire on such species is likely to be logistically impossible. Consequently, the effects of fire on forest and woodland fauna such as the spotted-tailed quoll are poorly understood, with many authors expressing concern that, potentially, wildfires are likely to be highly detrimental to resident quoll populations. The results of this study, however, concur with the few other studies in which forest mammal populations have been monitored before and after wildfire in suggesting that wildfires may not be as destructive to fauna as that imagined. The results of this work will provide information to assist in the preparation of management strategies for the species, such as recovery plans, as well as information for land managers preparing management plans, including fire management plans, for habitats in which spotted-tailed quolls are found throughout their range.

No description available.

The fetal lung epithelium secretes fluid into the potential pulmonary air-spaces by actively transporting chloride (Cl¯) into the lung lumen. This Cl¯-driven fluid secretion declines with the progression of lung development. Recent studies demonstrate that the A-type γ-aminobutyric acid receptor (GABAAR), a Cl¯ channel, and glutamic acid decarboxylase (GAD65/67), key GABA-synthesizing enzymes, are expressed in adult pulmonary epithelial cells (ECs), forming an autocrine GABAAR signaling system. My thesis study revealed that GABAAR π- and β2- subunits are expressed in high levels in the fetal rat lung epithelium and decline at birth, consistent with pattern of fluid secretion. Immunohistochemistry showed distinct profiles of expression for GABAAR subunits and GAD65/67. Treatment of alveolar ECs with dexamethasone reduced the GABAAR π-subunit expression. These results suggest that the GABAAR signaling in the fetal pulmonary epithelium is developmentally regulated and the GABAAR expression and GABAAR-mediated Cl¯ secretion in pulmonary ECs may be regulated by glucosteroids.

The fetal lung epithelium secretes fluid into the potential pulmonary air-spaces by actively transporting chloride (Cl¯) into the lung lumen. This Cl¯-driven fluid secretion declines with the progression of lung development. Recent studies demonstrate that the A-type γ-aminobutyric acid receptor (GABAAR), a Cl¯ channel, and glutamic acid decarboxylase (GAD65/67), key GABA-synthesizing enzymes, are expressed in adult pulmonary epithelial cells (ECs), forming an autocrine GABAAR signaling system. My thesis study revealed that GABAAR π- and β2- subunits are expressed in high levels in the fetal rat lung epithelium and decline at birth, consistent with pattern of fluid secretion. Immunohistochemistry showed distinct profiles of expression for GABAAR subunits and GAD65/67. Treatment of alveolar ECs with dexamethasone reduced the GABAAR π-subunit expression. These results suggest that the GABAAR signaling in the fetal pulmonary epithelium is developmentally regulated and the GABAAR expression and GABAAR-mediated Cl¯ secretion in pulmonary ECs may be regulated by glucosteroids.