Nosema ceranae, a microsporidian, has been given much attention in recent years as it has been linked with Colony Collapse Disorder (CCD), which leads to the sudden deaths of honey bee colonies. It has been described that many pathogenic organisms secrete virulence factors in order to hijack its host`s cellular functions, but in most cases the underlying mechanisms of this process still remains to be deciphered. Cornman et al. (2009) have identified in N. ceranae a list of putative effector proteins (called secretome) destined to be secreted into the host, and I have taken this list for further investigation using a bioinformatical and experimental approaches. The principal aim of this project was to generate a N. ceranae ORFeome for genes predicted to be secreted, elucidate the function of effector candidates important for N. ceranae biology and/or pathogenicity, as well as to investigate any interactions between N. ceranae proteins and its host utilising two eukaryotic model organisms, budding yeast, S. cerevisiae, and fruit fly, D. melanogaster. A library of S. cerevisiae strains expressing N. ceranae proteins was generated utilising the Gateway® technology, and phenotypic and localisation screens were undertaken to investigate the N. ceranae secretome. Two N. ceranae ORFs, NcORF-15 (NcORF-02039) and NcORF-16 (NcORF-01159) encoding a putative thioredoxin and a hexokinase, respectively, were subjected to yeast complementation assays in order to assess their catalytic activity. NcORF-15, the putative thioredoxin, was able to rescue the sensitive phenotype of S. cerevisiae Δtrx2 under oxidative stress, whereas NcORF-16, the putative hexokinase, did not complement YSH7.4-3C, a triple knockout lacking hexokinase activity. A third N. ceranae effector candidate NcORF-4 (NcORF-00654), a putative proteasome subunit, was investigated for its nuclear localisation and protein interactions in both S. cerevisiae and D. melanogaster.

Fitoestrógenos são compostos naturais de origem vegetal com atividade estrogênica. Estão sendo amplamente investigados para a prevenção de doenças crônicas, coronarianas, câncer de próstata e mama, na redução de riscos de osteoporose e alívio nos sintomas da menopausa. Entre os fitoestrógenos já utilizados encontram-se a daidzeína, genisteína e gliciteína em matrizes complexas como drogas e extratos vegetais, cápsulas e comprimidos, requerendo desenvolvimento e validação de metodologias para a determinação das isoflavonas. As metodologias propostas foram a cromatografia em camada delgada de alta eficiência (CCDAE) e a cromatografia líquida de alta eficiência (CLAE). A fase móvel acetato de etila:hexano (8:2 v/v) foi utilizada para determinação do perfil cromatográfico das isoflavonas agliconas daidzeína, gliciteína e genisteína e a fase móvel acetato de etila:tolueno:ácido fórmico (8: 1: 1 v/v/v) para isoflavonas glicosiladas e não glicosiladas por CCDAE. Para a determinação quantitativa das isoflavonas glicosiladas por CLAE foi proposta uma hidrólise ácida com HCl 3M e aquecimento em banho-maria durante uma hora, como tratamento prévio. A determinação analítica das isoflavonas daidzeína, genisteína, formononetina e biochanina A por CLAE, em modo isocrático foi realizada utilizando coluna cromatográfica monolítica Chromolith® RP-18, 100-4,6mm, fase móvel constituída por água:acetonitrila (6:4 v/v), vazão 0,6mL/min e detecção a 260nm. Os resultados obtidos mostraram linearidade com coeficiente de correlação de 0,9995 para daidzeína, 0,9996 para genisteína, 0,9997 para formononetina e 0.9999 para biochanina A. A precisão e a exatidão apresentaram resultados satisfatórios. Boa resolução e rápida separação dos fármacos em formulações farmacêuticas também foram obtidas. Portanto, pode ser usado nas análises de rotina nos laboratórios de controle de qualidade de fitoterápicos. / Phytoestrogens are natural compounds of plants with estrogenic activity. They are being widely investigated in the prevention of chronic coronary diseases, in prostate and breast cancer, in the reduction of osteoporosis risk and relief of menopause symptoms. Among phytoestrogens already in use are daidzein, genistein and glycitein and they are present in complex matrix such as phytopharmaceuticals, plant extracts, capsules and tablets. These preparations require development and validation of methodologies for quantitative determination of isoflavones. The proposed methodologies include high performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC). The HPTLC coupled with densitometry can be used for quantitative analysis. The mobile phase constituted of ethyl acetate:hexane (8:2 v/v) was used to determine chromatographic profiles of isoflavone aglycones, daidzein, glycitein and genistein. The mobile phase constituted of ethyl acetate:toluene:formic acid (8:1:1 v/v/v) was used for determination of isoflavones glycosides and non-glycosides. For the quantitative determination of isoflavone glycosides with HPLC, an acid hydrolysis with 3M HCl and heating in water-bath for an hour was proposed as sample pretreatment step. The analytic determination of isoflavones daidzein, genistein, formononetin and biochanin A using HPLC was accomplished. The chromatography was carried out in isocratic mode with Chromolith®, a monolithic RP-18 column, (100x4.6mm) with mobile phase constituted of water:acetonitrile (6:4 v/v) operated at a flow rate of 0.6mL/min and detection was made at 260nm. The results showed method linearity with correlation coefficient of 0.9995 to daidzein, 0.9996 for genistein, 0.9997 for formononetin and 0.9999 for biochanin A. The precision and accuracy data presented satisfactory results. Good resolution and faster separation of compounds in pharmaceutical formulations were also obtained. The proposed method can be used in the routine analyses of phytopharmaceuticals in quality control laboratories.

O fluoroacetato de sódio (FAS), também conhecido como \"composto 1080\", foi um raticida amplamente utilizado no combate de roedores a partir da década de 40, mas que teve seu uso proibido em diversos países, incluindo o Brasil, devido a sua alta toxicidade e ausência de tratamento efetivo nas intoxicações. A despeito deste fato, diversos casos de intoxicação ainda são registrados em seres humanos e animais, em virtude de seu uso ilícito como raticida doméstico ou adição em iscas nas intoxicações criminosas. O FAS, na natureza, pode ser encontrado, também, em algumas plantas tóxicas de grande importância no Brasil, como a Palicourea marcgravii, e possivelmente seja o princípio ativo tóxico de outras plantas de relevância, como a Mascagnia rigida. Neste sentido, este trabalho objetivou o estudo dos aspectos clínicos e anatomopatológicos encontrados nas intoxicações provocadas pelo FAS, em ratos, comparando estes com os encontrados nas intoxicações por extrato aquoso da M. rígida. Também foram comparadas metodologias analíticas para identificação deste agente tóxico utilizando cromatografia em camada delgada (CCD) e cromatografia de alta eficiência (CLAE) com dois tipos de detectores o de UV e o de Condutividade Iônica. Para tanto, quatro grupos contendo seis ratos Wistar, machos, receberam em dose única por via oral 7,0 mg/kg de FAS ou água destilada ou ainda 1,6 ou 3,2 g/kg de extrato aquoso de M. rígida, sendo avaliados os sinais clínicos apresentados por estes animais. Após eutanásia foram coletados fragmentos de cérebro, coração, fígado, rins, pulmão, intestino e baço para exame histopatológico. Tanto os animais que receberam FAS como os que receberam extrato aquoso de M. rígida apresentaram sintomatologia similar com presença de prostração, desconforto respiratório, tremores, pêlos arrepiados, cromodacriorreia, convulsão tônica e tônico-clônica. Na necropsia destes animais não foram observadas alterações macroscópicas, contudo alterações histopatológicas significativas foram encontradas no cérebro, miocárdio, fígado e rins. Os estudos das análise toxicológicas utilizando as três técnicas cromatográficas mostraram que os resultados obtidos na CCD foram adequados para detecção de FAS nas concentrações acima de 1,0 μg do padrão puro e as análises das duas plantas estudadas apresentaram resultados positivos para FAS. As técnicas utilizando CLAE, com dois diferentes detectores, mostraram que quando se utiliza detector de UV o limite de detecção é de 3,75 μg injetado ou de 75 μg/mL e foi possível, também, detectar a presença de FAS nas duas plantas estudadas, porém a técnica não se apresentou adequada para a detecção de intoxicação em tecido animal. O detector de condutividade iônica mostrou limite de detecção de 0,30 μg/mL, isto é, o mais sensível entre os três métodos estudados, e portanto o mais adequado para diagnóstico de intoxicação em tecido animal; no entanto, para identificação do FAS na M. rigida este método não se mostrou adequado devido a interferentes no extrato aquoso e no soro de animais intoxicados. / Sodium fluoroacetate (SFA), also known as \"compound 1080,\" was a rodenticide widely used in the 1940s, but its use was banned in several countries, including Brazil, for its high toxicity and lack of an effective treatment in poisoning. Despite this fact, several cases of poisoning are still registered in humans and animals, because of their misuse as baits, in addition to criminal poisoning. SFA, in the form of natural molecules, can be found in some toxic plants of great importance in Brazil, such as Palicourea marcgravii, and possibly is the active principle of other plants of relevance, such as Mascagnia rigid. Therefore, this study aimed to evaluate the clinical and pathological findings in poisoning caused by the SFA in rats, and compare them with those found in poisoning by the aqueous extract of M. rigid. Moreover, we compared analytical methods for identifying this toxic agent using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) employing two types of detectors: ultraviolet (UV) and ionic conductivity. Therefore, four groups containing six male Wistar rats each, received a single oral dose of 7.0 mg / kg of SFA, an equal volume of distilled water, or 1.6 or 3.2 g / kg of an aqueous extract of M. rigid, and the clinical signs displayed by these animals were evaluated. After euthanasia fragments of brain, heart, liver, kidney, lung, intestine and spleen were collected for histological examination. Both the animals that received SFA and those who received the aqueous extract of M. rigid showed similar symptoms, including prostration, respiratory distress, tremors, shivering, and tonic or tonic-clonic seizures. At necropsy, these animals did not display macroscopic changes. Nevertheless, significant histopathological changes were found in the brain, heart, liver and kidneys. Toxicological analysis using the three chromatographic techniques described above showed that the results obtained in the TLC were suitable for detection of SFA at concentrations above 1.0 μg of pure standard, and analysis of the two plants mentioned showed positive results for SFA. The techniques using HPLC with two different detectors showed that when using the UV detector, the detection limit is of 3.75 μg (injected) or 75 μg / mL, and SFA could also be detected by these methods in the two plants studied, but the technique does not seem suitable for the detection of intoxication in animal tissue samples. The ionic conductivity detector showed a detection limit of 0.30 μg/ mL, therefore the most sensitive of the three methods studied, and hence is the most appropriate for diagnosis of poisoning in animal tissue; however, for the identification of SFA in the M. rigid, this method was not appropriate due to a putative interference in the aqueous extract.

Doctor of Philosophy (PhD) / This thesis is an enquiry into the social and political role, in Australia, of practices that have attracted such labels as ‘community arts’, ‘cultural animation’, ‘cultural action’, or ‘community cultural development’ (CCD). It is often argued that such practices offer an effective means to bring about social and political change for people and communities who participate in them. Looking specifically at theatre-based approaches to CCD in Australia, this thesis examines an alternative hypothesis, namely that such projects and programs can contribute to the continued marginalisation of those who take part in them. Using a combination of Pierre Bourdieu’s theoretical approach to field analysis, Don Handelman’s analytical framework of special events and Baz Kershaw’s theory of potential efficacy, I carry out an ethnographic and performance-based analysis of a particular project called The Longest Night (TLN), which was devised in collaboration with young people from The Parks, a cluster of suburbs north west of Adelaide, South Australia, and in collaboration between Urban Theatre Projects, a small Sydney-based theatre company with a reputation for doing socially and politically challenging work, young people living in The Parks and local partner organisations, for the 2002 Adelaide Festival. I find that in some instances participation in CCD projects and programs is an enabling factor, creating change opportunities in cultural, economic and/or political spheres in the lives of those who take part, whilst at other times it is a constraining factor. Participation in CCD projects and programs creates possibilities because the practices are potentially subversive and foster elements of learning and change in some participants. It also creates limitations because CCD practitioners operate within a subfield of social and cultural practices where the mechanisms and structures in place, indirectly, tend to help reproduce legitimised social and cultural values and norms.

L'étude des atomes exotiques, où un électron de l'atome est remplacé par une autre particule de charge négative peut fournir des informations sur la particule ou sur le noyau, ou encore sur l'interaction entre la particule et le noyau. <br /><br />Au cours de ce travail, nous nous sommes intéressés, d'une part, aux atomes pioniques dans le but de déterminer la masse du pion négatif avec une très bonne précision (4 ppm), et d'autre part, aux atomes antiprotoniques pour étudier l'interaction forte entre un nucléon et un antinucléon. A cet égard, un spectromètre de Bragg de haute résolution et une trappe cyclotron ont été utilisés. Avec sa capacité de fabriquer en grande quantité les atomes exotiques, la trappe cyclotron a fourni une source intense de rayons-X. Les cristaux courbes du spectromètre focalisant les rayons-X ont assuré un gain considérable en intensité. Grâce à leur résolution spatiale, les CCD utilisés comme détecteurs ont permis une détermination précise de l'angle de Bragg correspondant à la transition à mesurer.<br /><br />L'utilisation d'un milieu gazeux pour arrêter les pions a résolu l'ambiguïté provenant de l'hypothèse sur le nombre d'électrons K recapturés de la cible. D'un autre côté, nous avons réalisé la première mesure directe du décalage et de l'élargissement hadroniques du niveau 2p dans l'hydrogène antiprotonique.

<p>In the newspaper printing industry, offset is the dominating printing method and the use of multicolour printing has increased rapidly in newspapers during the last decade. The offset printing process relies on the assumption that an uniform film of ink of right thickness is transferred onto the printing areas. The quality of reproduction of colour images in offset printing is dependent on a number of parameters in a chain of steps and in the end it is the amount and the distribution of ink deposited on the substrate that create the sensation and thus the perceived colours. We identify three control points in the offset printing process and present methods for assessing the printing process quality in two of these points:</p><p>• Methods for determining if the printing plates carry the correct image</p><p>• Methods for determining the amount of ink deposited onto the newsprint</p><p>A new concept of colour impression is introduced as a measure of the amount of ink deposited on the newsprint. Two factors contribute to values of the colour impression, the halftone dot-size and ink density. Colour impression values are determined on gray-bars using a CCD-camera based system. Colour impression values can also be determined in an area containing an arbitrary combination of cyan magenta and yellow inks. The correct amount of ink is known either from a reference print or from prepress information. Thus, the deviation of the amount of ink can be determined that can be used as control value by a press operator or as input to a control system.</p><p>How a closed loop controller can be designed based on the colour impression values is also shown.</p><p>It is demonstrated that the methods developed can be used for off-line print quality monitoring and ink feed control, or preferably in an online system in a newspaper printing press.</p> / Report code: LiU-TEK-LIC-2005:02.

<p>The X-ray Multi-mirror Mission, XMM-Newton was launched by the European Space Agency, ESA, in 1999. XMM-Newton carries six cameras, including a silicon pn-junction Charge Coupled Device, or pn-CCD camera. This camera has six operating modes, spatially as well as time resolved. The main objective of this project is to refine the Burst mode energy correction in order to align the measured energy spectra observed in the Burst mode with the spectra taken in the Full Frame mode. An observation of the line-rich supernova remnant called Cassiopeia A is used to evaluate the line positions in each mode such that the energy correction function used for the alignment can be modified accordingly. The analysis further treats the application of the correction on a source with a continuous spectrum, the Crab nebula. Discussion shows how to reduce eventual residuals in the Crab spectrum by modifying the correction function while keeping the alignment of the Cas-A spectra. The final product is an update of the corresponding published calibration file.</p>

This thesis will present the design, implementation, and testing of a high voltage Charge-Coupled Device (CCD) controller ASIC for the Large Synoptic Survey Telescope (LSST), which will be used to study dark energy and dark matter. The LSST observatory, which includes a 3.2-gigapixel camera, will cover the entire sky every three nights by taking continuous 15-second exposures. The CCD controller ASIC, or Sensor Control Chip (SCC), will provide five CCD driver channels that are capable of generating serial or parallel clock signals for the LSST’s imaging sensors during readout mode. The SCC will also provide three programmable bias voltages for the CCDs along with eight supplementary programmable voltages and currents for the CCD’s output drain terminals. Additionally, the controller ASIC includes eight control signals for a separate Analog Signal Processing Integrated Circuit (ASPIC) that is designed as the readout chip for LSST. The SCC is designed to operate down to 153 K. Fabricated in a commercially available 0.8-micron Bipolar-CMOS-DMOS Silicon-On-Insulator (BCD-SOI) process, the SCC has been verified to meet all design requirements.

The X-ray Multi-mirror Mission, XMM-Newton was launched by the European Space Agency, ESA, in 1999. XMM-Newton carries six cameras, including a silicon pn-junction Charge Coupled Device, or pn-CCD camera. This camera has six operating modes, spatially as well as time resolved. The main objective of this project is to refine the Burst mode energy correction in order to align the measured energy spectra observed in the Burst mode with the spectra taken in the Full Frame mode. An observation of the line-rich supernova remnant called Cassiopeia A is used to evaluate the line positions in each mode such that the energy correction function used for the alignment can be modified accordingly. The analysis further treats the application of the correction on a source with a continuous spectrum, the Crab nebula. Discussion shows how to reduce eventual residuals in the Crab spectrum by modifying the correction function while keeping the alignment of the Cas-A spectra. The final product is an update of the corresponding published calibration file.

The process of laser cladding consists of depositing successive layers of molten metallic powder to create a near-net shape. A high-power laser is used to melt incoming metallic powder, which forms a melt pool on the surface. As the latter moves beneath the laser, this newly created melt pool solidifies. By properly controlling the trajectory of deposition tracks, one can create a diverse range of shapes with varying complexities. However, the process is very sensitive to parameters, requiring constant attention from technicians. This lends itself perfectly to the addition of automatic controllers whereby supervision is minimal. <br /><br /> In this thesis, an optical sensor is developed to monitor the process zone. The sensor will output a measurement of the height of solidified clad, which in turn can be used by a controller to adjust this geometrical feature. The thesis is divided into three main parts, each contributing to the final algorithm. <br /><br /> First, in Chapter 3 an analysis is performed on the light irradiating from the interaction zone (or melt pool). It is stated that the dominating source of light is governed by blackbody radiation from this molten metal. This is confirmed by analyzing a series of images captured through a digital camera, where various narrow bandpass filters were utilized to selectively view a portion of the CCD-sensor's spectrum. This investigation also leads to the selection of bandpass filter such that a stable, relatively intense melt pool is captured through the digital camera's CCD-sensor. <br /><br /> Second, in Chapter 4 the captured images are taken through a pair of image processing techniques, outputting a series of coordinates representating the melt pool's boundary. The image is first analyzed to calculate an optimal threshold level based on the minimization of fuzzy entropy. With this threshold selected, the grayscale image is converted into black-and-white, where the white pixels represent the melt pool. After this step, the melt pool's boundary is extracted through an 8-connectivity border tracing algorithm. This technique outputs a series of coordinates (in pixels) as though one were traveling along the melt pool in a clockwise rotation. <br /><br /> Last, Chapter 5 analyzes these pixel coordinates to extract the melt pool's height. The coordinates are first transformed into real-world coordinates, by use of a perspective transformation. This transformation essentially yields the melt pool's shadow, as created by a light-source coincident with the camera. As a result, the melt pool's height is estimated based upon a right-angle triangle, where the camera's angle is known, and the projected coordinates represent the shadow length (triangle's base). <br /><br /> The result of applying this series of steps to the estimation of clad heights is found at the end of Chapter 5. Results varied dramatically, from 4% error to 393%. Although the errors are large at times, they are mainly caused by a bias in the estimate. That is, the dynamics of the true clad formation are very well predicted by the algorithm, however, shifting by a certain amount. This amount varies both with substrate velocity, and the clad's direction of travel, relative to the camera. A partial explanation is given such that the clad's height is offset from the laser center-point, which is a function of both these parameters. However, the specific relationship requires further experimentation.