998 resultados para System Circle
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O presente artigo busca estabelecer o local e a função do capital portador de juros em O Capital, visando com isso jogar luz sobre a estrutura eminentemente sistemática desta obra. Deste modo a obra máxima de Marx aparecerá muito próxima do idealismo alemão (particularmente de Fichte e Hegel), onde a questão da sistematicidade (da filosofia) foi sentida de maneira mais premente.
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Banana bunchy top is regarded as the most important viral disease of banana, causing significant yield losses worldwide. The disease is caused by Banana bunchy top virus (BBTV), which is a circular ssDNA virus belonging to the genus Babuvirus in the family Nanoviridae. There are currently few effective control strategies for this and other ssDNA viruses. “In Plant Activation” (InPAct) is a novel technology being developed at QUT for ssDNA virus-activated suicide gene expression. The technology exploits the rolling circle replication mechanism of ssDNA viruses and is based on a unique “split” gene design such that suicide gene expression is only activated in the presence of the viral Rep. This PhD project aimed to develop a BBTV-based InPAct system as a suicide gene strategy to control BBTV. The BBTV-based InPAct vector design requires a BBTV intergenic region (IR) to be embedded within an intron in the gene expression cassette. To ensure that the BBTV IR would not interfere with intron splicing, a TEST vector was initially generated that contained the entire BBTV IR embedded within an intron in a β-glucuronidase (GUS) expression vector. Transient GUS assays in banana embryogenic cell suspensions indicated that cryptic intron splice sites were present within the IR. Transcript analysis revealed two cryptic intron splice sites in the Domain III sequence of the CR-M within the IR. Removal of the CR-M from the TEST vector resulted in an enhancement of GUS expression suggesting that the cryptic intron splice sites had been removed. An InPAct GUS vector was subsequently generated that contained the modified BBTV IR, with the CR-M (minus Domain III) repositioned within the InPAct cassette. Using transient histochemical and fluorometric GUS assays in banana embryogenic cells, the InPAct GUS vector was shown to be activated in the presence of the BBTV Rep. However, the presence of both BBTV Rep and Clink was shown to have a deleterious effect on GUS expression suggesting that these proteins were cytotoxic at the levels expressed. Analysis of replication of the InPAct vectors by Southern hybridisation revealed low levels of InPAct cassette-based episomal DNA released from the vector through the nicking/ligation activity of BBTV Rep. However, Rep-mediated episomal replicons, indicative of rolling circle replication of the released circularised cassettes, were not observed. The inability of the InPAct cassette to be replicated was further investigated. To examine whether the absence of Domain III of the CR-M was responsible, a suite of modified BBTV-based InPAct GUS vectors was constructed that contained the CR-M with the inclusion of Domain III, the CR-M with the inclusion of Domain III and additional upstream IR sequence, or no CR-M. Analysis of replication by Southern hybridisation revealed that neither the presence of Domain III, nor the entire CR-M, had an effect on replication levels. Since the InPAct cassette was significantly larger than the native BBTV genomic components (approximately 1 kb), the effect of InPAct cassette size on replication was also investigated. A suite of size variant BBTV-based vectors was constructed that increased the size of a replication competent cassette to 1.1 kbp through to 2.1 kbp.. Analysis of replication by Southern hybridisation revealed that an increase in vector size above approximately 1.5 - 1.7 kbp resulted in a decrease in replication. Following the demonstration of Rep-mediated release, circularisation and expression from the InPAct GUS vector, an InPAct vector was generated in which the uidA reporter gene was replaced with the ribonuclease-encoding suicide gene, barnase. Initially, a TEST vector was generated to assess the cytotoxicity of Barnase on banana cells. Although transient assays revealed a Barnase-induced cytotoxic effect in banana cells, the expression levels were sub-optimal. An InPAct BARNASE vector was generated and tested for BBTV Rep-activated Barnase expression using transient assays in banana embryogenic cells. High levels of background expression from the InPAct BARNASE vector made it difficult to accurately assess Rep-activated Barnase expression. Analysis of replication by Southern hybridisation revealed low levels of InPAct cassette-based episomal DNA released from the vector but no Rep-mediated episomal replicons indicative of rolling circle replication of the released circularised cassettes were again observed. Despite the inability of the InPAct vectors to replicate to enable high level gene expression, the InPAct BARNASE vector was assessed in planta for BBTV Rep-mediated activation of Barnase expression. Eleven lines of transgenic InPAct BARNASE banana plants were generated by Agrobacterium-mediated transformation and were challenged with viruliferous Pentalonia nigronervosa. At least one clonal plant in each line developed bunchy top symptoms and infection was confirmed by PCR. No localised lesions were observed on any plants, nor was there any localised GUS expression in the one InPAct GUS line challenged with viruliferous aphids. The results presented in this thesis are the first study towards the development of a BBTV-based InPAct system as a Rep-activatable suicide gene expression system to control BBTV. Although further optimisation of the vectors is necessary, the preliminary results suggest that this approach has the potential to be an effective control strategy for BBTV. The use of iterons within the InPAct vectors that are recognised by Reps from different ssDNA plant viruses may provide a broad-spectrum resistance strategy against multiple ssDNA plant viruses. Further, this technology holds great promise as a platform technology for the molecular farming of high-value proteins in vitro or in vivo through expression of the ssDNA virus Rep protein.
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Despite various approaches, the production of biodegradable plastics such as polyhydroxybutyrate (PHB) in transgenic plants has met with limited success due largely to low expression levels. Even in the few instances where high levels of protein expression have been reported, the transgenic plants have been stunted indicating PHB is phytotoxic (Poirier 2002). This PhD describes the application of a novel virus-based gene expression technology, termed InPAct („In Plant Activation.), for the production of PHB in tobacco and sugarcane. InPAct is based on the rolling circle replication mechanism by which circular ssDNA viruses replicate and provides a system for controlled, high-level gene expression. Based on these features, InPAct was thought to represent an ideal system to enable the controlled, high-level expression of the three phb genes (phbA, phbB and phbC) required for PHB production in sugarcane at a preferred stage of plant growth. A Tobacco yellow dwarf virus (TbYDV)-based InPAct-phbA vector, as well as linear vectors constitutively expressing phbB and phbC were constructed and different combinations were used to transform tobacco leaf discs. A total of four, eight, three and three phenotypically normal tobacco lines were generated from discs transformed with InPAct-phbA, InPAct-phbA + p1300-TaBV P-phbB/phbC- 35S T, p1300-35S P-phbA-NOS T + p1300-TaBV P-phbB/phbC-35S T and InPAct-GUS, respectively. To determine whether the InPAct cassette could be activated in the presence of the TbYDV Rep, leaf samples from the eight InPActphbA + p1300-TaBV P-phbB/phbC-35S T plants were agroinfiltrated with p1300- TbYDV-Rep/RepA. Three days later, successful activation was indicated by the detection of episomes using both PCR and Southern analysis. Leaf discs from the eight InPAct-phbA + p1300-TaBV P-phbB/phbC-35S T transgenic plant lines were agroinfiltrated with p1300-TbYDV-Rep/RepA and leaf tissue was collected ten days post-infiltration and examined for the presence of PHB granules. Confocal microscopy and TEM revealed the presence of typical PHB granules in five of the eight lines, thus demonstrating the functionality of InPActbased PHB production in tobacco. However, analysis of leaf extracts by HPLC failed to detect the presence of PHB suggesting only very low level expression levels. Subsequent molecular analysis of three lines revealed low levels of correctly processed mRNA from the catalase intron contained within the InPAct cassette and also the presence of cryptic splice sites within the intron. In an attempt to increase expression levels, new InPAct-phb cassettes were generated in which the castorbean catalase intron was replaced with a synthetic intron (syntron). Further, in an attempt to both increase and better control Rep/RepA-mediated activation of InPAct cassettes, Rep/RepA expression was placed under the control of a stably integrated alc switch. Leaf discs from a transgenic tobacco line (Alc ML) containing 35S P-AlcR-AlcA P-Rep/RepA were supertransformed with InPAct-phbAsyn or InPAct-GUSsyn using Agrobacterium and three plants (lines) were regenerated for each construct. Analysis of the RNA processing of the InPAct-phbAsyn cassette revealed highly efficient and correct splicing of the syntron, thus supporting its inclusion within the InPAct system. To determine the efficiency of the alc switch to activate InPAct, leaf material from the three Alc ML + InPAct-phbAsyn lines was either agroinfiltrated with 35S P-Rep/RepA or treated with ethanol. Unexpectedly, episomes were detected not only in the infiltrated and ethanol treated samples, but also in non-treated samples. Subsequent analysis of transgenic Alc ML + InPAct-GUS lines, confirmed that the alc switch was leaky in tissue culture. Although this was shown to be reversible once plants were removed from the tissue culture environment, it made the regeneration of Alc ML + InPAct-phbsyn plant lines extremely difficult, due to unintentional Rep expression and therefore high levels of phb expression and phytotoxic PHB production. Two Alc ML + InPAct-phbAsyn + p1300-TaBV P-phbB/phbC-35S T transgenic lines were able to be regenerated, and these were acclimatised, alcohol-treated and analysed. Although episome formation was detected as late as 21 days post activation, no PHB was detected in the leaves of any plants using either microscopy or HPLC, suggesting the presence of a corrupt InPAct-phbA cassette in both lines. The final component of this thesis involved the application of both the alc switch and the InPAct systems to sugarcane in an attempt to produce PHB. Initial experiments using transgenic Alc ML + InPAct-GUS lines indicated that the alc system was not functional in sugarcane under the conditions tested. The functionality of the InPAct system, independent of the alc gene switch, was subsequently examined by bombarding the 35S Rep/RepA cassette into leaf and immature leaf whorl cells derived from InPAct-GUS transgenic sugarcane plants. No GUS expression was observed in leaf tissue, whereas weak and irregular GUS expression was observed in immature leaf whorl tissue derived from two InPAct- GUS lines and two InPAct-GUS + 35S P-AlcR-AlcA P-GUS lines. The most plausible reason to explain the inconsistent and low levels of GUS expression in leaf whorls is a combination of low numbers of sugarcane cells in the DNA replication-conducive S-phase and the irregular and random nature of sugarcane cells bombarded with Rep/RepA. This study details the first report to develop a TbYDV-based InPAct system under control of the alc switch to produce PHB in tobacco and sugarcane. Despite the inability to detect quantifiable levels of PHB levels in either tobacco or sugarcane, the findings of this study should nevertheless assist in the further development of both the InPAct system and the alc system, particularly for sugarcane and ultimately lead to an ethanol-inducible InPAct gene expression system for the production of bioplastics and other proteins of commercial value in plants.
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Listening comprehension is the primary channel of learning a language. Yet of the four dominant macro-skills (listening, speaking, reading and writing), it is often difficult and inaccessible for second and foreign language learners due to its implicit process. The secondary skill, speaking, proceeds listening cognitively. Aural/oral skills precede the graphic skills, such as reading and writing, as they form the circle of language learning process. However, despite the significant relationship with other language skills, listening comprehension is treated lightly in the applied linguistics research. Half of our daily conversation and three quarters of classroom interaction are virtually devoted to listening comprehension. To examine the relationship of listening skill with other language skills, the outcome of 1800 Iranian participants undertaking International English Language Testing System (IELTS) in Tehran indicates the close correlation between listening comprehension and the overall language proficiency.
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Short-duration (5- or 10-day) deployments of pop-up satellite archival tags were used to estimate survival of white marlin (Tetrapturus albidus) released from the western North Atlantic recreational fishery. Forty-one tags, each recording temperature, pressure, and light level readings approximately every two minutes for 5-day tags (n= 5) or four minutes for 10-day tags (n= 36), were attached to white marlin caught with dead baits rigged on straight-shank (“J”) hooks (n =21) or circle hooks (n=20) in offshore waters of the U.S. Mid-Atlantic region, the Dominican Republic, Mexico, and Venezuela. Forty tags (97.8%) transmitted data to the satellites of the Argos system, and 33 tags (82.5%) transmitted data consistent with survival of tagged animals over the deployment duration. Approximately 61% (range: 19−95%) of all archived data were successfully recovered from each tag. Survival was significantly (P<0.01) higher for white marlin caught on circle hooks (100%) than for those caught on straight-shank (“J”) hooks (65%). Time-to-death ranged from 10 minutes to 64 hours following release for the seven documented mortalities, and five animals died within the first six hours after release. These results indicate that a simple change in hook type can significantly increase the survival of white marlin released from recreational fis
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An effective face detection system used for detecting multi pose frontal face in gray images is presented. Image preprocessing approaches are applied to reduce the influence of the complex illumination. Eye-analog pairing and improved multiple related template matching are used to glancing and accurate face detecting, respectively. To shorten the time cost of detecting process, we employ prejudge rules in checking candidate image segments before template matching. Test by our own face database with complicated illumination and background, the system has high calculation speed and illumination independency, and obtains good experimental results.
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In this paper we present a robust face location system based on human vision simulations to automatically locate faces in color static images. Our method is divided into four stages. In the first stage we use a gauss low-pass filter to remove the fine information of images, which is useless in the initial stage of human vision. During the second and the third stages, our technique approximately detects the image regions, which may contain faces. During the fourth stage, the existence of faces in the selected regions is verified. Having combined the advantages of Bottom-Up Feature Based Methods and Appearance-Based Methods, our algorithm performs well in various images, including those with highly complex backgrounds.
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Huazhong Univ Sci & Technol, Natl Tech Univ Ukraine, Huazhong Normal Univ, Harbin Inst Technol, IEEE Ukraine Sect, I& M/CI Joint Chapter
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A feature detection system has been developed for real-time identification of lines, circles and people legs from laser range data. A new method sutable for arc/circle detection is proposed: the Inscribed Angle Variance (IAV). Lines are detected using a recursive line fitting method.
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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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Over the past decade or two, restorative justice has become a popular approach for the criminal justice system to take in Canada, New Zealand, and Australia. In part, this is due in all three countries to an appalling disproportionality in the incarceration rates for racialized minorities. As the authors of "Will the Circle Be Unbroken?" point out, however, governments have been attracted to restorative justice for cost-cutting reasons as well. A burning question, therefore, is whether restorative justice works.
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We have done a new analysis of the available observations of the GJ581 exoplanetary system. Today this system is controversial due to choices that can be done in the orbital determination. The main ones are the occurrence of aliases and the additional bodies-the planets f and g-announced in Vogt et al. (Astrophys J 723:954-965, 2010). Any dynamical study of exoplanets requires the good knowledge of the orbital elements and the investigations involving the planet g are particularly interesting, since this body would lie in the habitable zone (HZ) of the star GJ581. This region, for this system, is very attractive of the dynamical point of view due to several resonances of two and three bodies present there. In this work, we investigate the conditions under which the planet g may exist. We stress the fact that the planet g is intimately related with the orbital elements of the planet d; more precisely, we conclude that it is not possible to disconnect its existence from the determination of the eccentricity of the planet d. Concerning the planet f, we have found one solution with period a parts per thousand 450 days, but we are judicious about any affirmation concerning this body because its signal is in the threshold of detection and the high period is in a spectral region where the occurrence of aliases is very common. Besides, we outline some dynamical features of the HZ with the dynamical map and point out the role played by some resonances laying there.
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We report the discovery of two low-mass companions to the young A0V star HD 1160 at projected separations of 81 +/- 5 AU (HD 1160 B) and 533 +/- 25 AU (HD 1160 C) by the Gemini NICI Planet-Finding Campaign. Very Large Telescope images of the system taken over a decade for the purpose of using HD 1160 A as a photometric calibrator confirm that both companions are physically associated. By comparing the system to members of young moving groups and open clusters with well-established ages, we estimate an age of 50(-40)(+50) Myr for HD 1160 ABC. While the UVW motion of the system does not match any known moving group, the small magnitude of the space velocity is consistent with youth. Near-IR spectroscopy shows HD 1160 C to be an M3.5 +/- 0.5 star with an estimated mass of 0.22(-0.04)(+0.03) M-circle dot, while NIR photometry of HD 1160 B suggests a brown dwarf with a mass of 33(-9)(+12) M-Jup. The very small mass ratio (0.014) between the A and B components of the system is rare for A star binaries, and would represent a planetary-mass companion were HD 1160 A to be slightly less massive than the Sun.