300 resultados para Danner, Harley


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Acknowledgements BP Exploration Co. is thanked for funding, and particularly the Carbonate Team (Anna Matthews, Teresa Sabato Ceraldi, and Darryl G. Green) for supporting this research and for fruitful discussions. Mark Anderson, Kim Rosewell, and Tony Sinclair (University of Hull) are thanked for laboratory assistance, and for SEM sample preparation and set-up respectively. The technical and human support from Prof. Jörg Hardege and Maggy A. Harley (University of Hull) was key to perform these experiments. We would like to acknowledge an anonymous reviewer for the detailed and constructive comments, and Brian Jones's editorial handling of the manuscript which is greatly appreciated.

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The jabuticaba fruit tree from classified in the Myrtaceae family and Plinia genre. There are about nine species of this fruit tree, that include as most important, Plinia trunciflora (jabuticaba de cabinho), naturally occurring in southwestern Paraná State, Brazil, P. cauliflora (jabuticaba Paulista or Jabuticaba Açu) and P. jaboticaba (Vell) (jabuticaba sabará), with all the over species producing fruit for the industry or fresh consumption. Nevertheless, there aren‟t commercial orchards with this culture, with highest yield part from extractive. This fact can be combined with lack of technical knowledge for the plants produce in the field. As these species are found in the forest, the first point is whether they can adapt to other light intensity conditions. The aim of this work was to identify the adaptive behavior of jabuticaba fruit seedling and tree when they were put in different light intensities and what this can be considered ideal for the growth, as well as, its influence in the leaves secondary compounds production. Two experiments were conducted, with the first involved with the study of the seedlings and the second with plants in the field. The work was carried out at Universidade Tecnológica Federal do Paraná – Câmpus Dois Vizinhos, Paraná State - Brazil. The experimental design was a completely randomized and a block design with four treatments and four replications of 10 seedlings or two plants per plot, according to nursery or orchard conditions, respectively. The treatments were base according to the light intensity. The treatments used were, 1 - full sun, similar the orchard condition, with 0% shading; 2 - side cover with shade cloth and top with transparent plastic, representing a gap forest condition; 3 - side and top cover with shade cloth, representing stage where the forest canopy is closing, focusing only indirect sunlight; 4 - side and top cover with shade cloth, simulating a closed canopy condition, with PPD (photon flux density) of 10% (90% shading); 5 - side and top cover with shade cloth, simulating a more open canopy condition with PPD 65% (35% shading). The growth and development seedling and plant characteristics were evaluated once by month, as also, during time part in the plants the secondary metabolites leaves, soil activity microbiological and the fresh and dry matter root and shoot and, root length from seedlings. For the growth and development of jabuticaba Açú Paulista seedling recommend to use of side cover with shade cloth and top with transparent plastic, representing a gap forest condition. In orchard, for the growth and development of plants jabuticaba Híbrida tree it was recommended the use of side and top cover with shade cloth of some type. For production of secondary metabolites of leaves, the plant must to be full sunlight condition orchard.

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To design strategies for the conservation and use of genetic resources of tree species such as jaboticaba tree, it is essential to make the characterization. In southwestern Paraná region, there are several forest fragments containing native jaboticaba tree (Plinia cauliflora), whose materials have broad potential for commercial orchards or breeding programs. As is the potential genetic diversity of a population to produce different genotypes, it would be able to start in such a characterization one of these fragments. The aim was to characterize fruits of jaboticaba tree (P. caulifora) of forest fragment kept in Clevelândia - PR for the presence of phenotypic variability, seeking to identify those superiors named for future selection as farming or male parent, as well as estimate genetic divergence between them, as a complementary tool for this purpose. Also, verify the regeneration and spatial distribution of the species. For the study was defined portion of a hectare (10.000 m²), with all individuals identified, mapped, with local coordinate system, and measured height and diameter. Fruits were characterized by sensory and biochemical characteristics in two years, 70 genotypes at 2013 and 56 at 2014, and of these 33 genotypes in both years. As a pre-selection criteria was adopted the choice of 20% of the genotypes that showed the highest frequency of superiority in the evaluated characteristics of the fruit. Genetic divergence among 33 genotypes per year was analyzed. The distribution pattern and spatial association was evaluated by Ripley's K function. It was classified for the first time the following ontogenetic stages of jaboticaba tree, by plant height, seedling (from 0.01 to 0.99 m), juvenile (1.0 to 4.99 m), immature (> 5.0 m, non-reproductive), adult (reproductive). It was also have been describe for the first time the naturally occurring juxtaposed seedlings, indicating polyembryony. The number of regenerating identified in the population (seedlings: n = 2163; juveniles: n = 330; immature: n = 59) was much larger than the number of adults (n = 132). The species showed reverse J-shaped size structure standard, with high concentration of regenerating. The regeneration distribution occurs in aggregate pattern and there is seedling-adult dependence, due seed dispersal and seedling emergence closest to mothers. The jaboticaba tree regeneration is sufficient to maintain the species for long term in this population, which should serve as reference to regeneration success for other studies of this important fruiting species from Ombrofile Mixed Forests. Has been pre-selected the jaboticaba trees 7, 42, 43, 47, 54, 91, 97, 104, 105, 118, 134, 153, 154, 157, 163, 169, 177, 186, 212, J7-01 and J7- 02, and 16 and 194 the ones that can now be selected by the superior characteristics of both cycles. It was recommended to carry out hybridization between genotypes 79 and 119, and 96 to 148. The quality of fruit analyzed showed potential for use as a dual purpose serving both in natura market or processing.

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The most native fruit trees are belonging to Myrtaceae family, which it have as main marketing potential their fruit. Despite the wide acceptance of the fruits of these native fruit cultura, the establishment of commercial orchards is still necessary, because if it prevails extraction in the forest. To start the cultivo in the orchard, the first point is on the mother plant choice, which should provide superior characteristics when compared to other genotypes. Then, it is necessary to choose the method to can produce satisfactory amount of seedlings and preferably without it to lose the mother plant characteristics. For this, it adopts the asexual thechniques, with option for grafting, cuttings and air layering. These techniques when tested with native fruits tree, it proved limiting in theses results, with this, it should to test other it to recommend its use, especially, those fruit native of higher potential as jabuticaba tree, pitanga tree, sete capote tree and araça amarelo tree. The aim of this study was to test the use of asexual propagation through mini-cuttings in these native fruit trees, according to the time of collection, the mini-cutting length and concentration of IBA, as well as, it to relate the results of rooting with tryptophan extracted at certain times. The work was carried out at Universidade Tecnológica Federal do Paraná – Câmpus Dois Vizinhos, Brazil. The samples were collected each two months. The mini-cutting were prepared with 6 or 8 cm, with a pair of leaves reduced to 25% of the original size. The mini-cuttings had their base immersed in liquid solution of indole-butyric acid (IBA) in the concentrations of 0, 3000 and 6000 mg L-1 and then were placed in tubes containing commercial substrate. The experimental design was completely randomized with factorial 2 x 3 x 6 (mini-cutting length x IBA concentration x time of collection), with four replications, it being each plot varied according to the amount of shoots obtained by period time. After 120 days, the rooting and callus formation (%), average number of roots per mini-cutting and the average length of the roots were evaluated. After 60 days of these evaluations, the survival of mini-cuttings rooted after transplant was evaluated. It was evaluated also the production of mini-cuttings of each size in each period time. At the end of the experiment it was evaluated the percentage of survival of mother plantlets. For analysis of tryptophan was used materials branches, leaves and twigs with leaves, taken from the materials used for the production of mini-cutting. It was recommended for hybrid jabuticaba tree the use mini-cutting with eight cm, treated with 6000 mg L-1 of IBA and collected in June. For jabuticaba tree of cabinho and araça amarelo tree the period for propagation by mini-cuttings should be in August, regardless of IBA concentration and length of the mini-cutting. In the jabuticaba tree sabara and sete capote tree is important to obtain more satisfactory results realized the collect in October or December, with the same independence of other levels tested in other factors. However, for sete capote tree should test other techniques to increase the efficiency of propagation. And with pitanga tree recommended to the collection in June, but with 6cm the application of 3000 mg L-1 of IBA and 8 cm with 6000 mg L-1 of IBA.

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The instability of environment between years in climates of subtropical regions difficult to obtain peach trees genotypes with wide adaptation and stable production, contributing to poor crop. The climate instability can affect development stages as flower bud and vegetative bud formation. The factors understanding that control the bud formation, presents elementary importance for effective solutions search to these problems. The objective this work is verify the temperature effect, relative humidity and rainfall on bud density and length shoot (Brindilas) and identify genotypes with more adaptability and stability for this character. Was used 12 peach trees genotypes growing in experimental orchard in the Technology Federal of Paraná State University, Campus Pato Branco with Cfa Köppen climate according to the classification. Data of rainfall, hourly temperature were collected by the weather station of Simepar. They were used three plants for genotype (rehearsal), identify five shoots per tree, in May of each year. Were carried analyzes of length shoot CR (cm), count number of flower bud (GF) and vegetative bud (GV). Also calculated the relationship between GF/GV and flower bud density and vegetative bud density. Evaluations were performer annual 2007-2014. With these data adaptability and stability analyzes were performed using Biplot methodology and correlations analyzes (Pearson) with climates variables. They used the weather data to calculate the sums of hours with temperatures below 20 °C, temperatures between 20-25 °C, temperature between 25-30 °C and temperature above 30 °C, considering the period of August 1fst of the previous period to February 28 of the following year. Pearson correlation coefficients were used for path analysis, GF and DGF as basic variables. For CR, GV and GF the highest average occurred in 2009/10 period. The genotypes ‘BRS Kampai’ and ‘BRS Libra’ highest CR. They are considered stable and adapted as the CR genotypes ‘Casc. 967’ and ‘BRS Kampai’. There was negative correlation between CR and GV for Σh <20 ° C, Σh> 30 °C and Σh with URA <50% and positive correlation between these variables and Σh 25-30 °C and Σh with URA> 70%. The evaluation of GV ‘Cons. 681’ and ‘Casc. 1055’ can be considered adapted and stable. The lowest average was presented by the genotype ‘Sta. Áurea’ though the genotype is also stable. In GF evaluation genotypes are considered adapted ‘BRS Bonão’, ‘Casc. 1055’, ‘Cons. 681’ with adaptability to all evaluated period. In path analysis was direct effect Σh 25-30 °C on flower bud density. In evaluating DGV and DGF and the variations are due to genetic effect. The most adapted and stable genotypes for DGV were ‘T. Beauty’, ‘T. Snow’, ‘Casc. 1055’ and ‘Cons. 681’. CR and GV variables are strongly affected by environment. GF is strongly affected by genetic conditions and moderately affected by environment. DGV and DGF are affected basically by genetic conditions.

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Membrane proteins, which reside in the membranes of cells, play a critical role in many important biological processes including cellular signaling, immune response, and material and energy transduction. Because of their key role in maintaining the environment within cells and facilitating intercellular interactions, understanding the function of these proteins is of tremendous medical and biochemical significance. Indeed, the malfunction of membrane proteins has been linked to numerous diseases including diabetes, cirrhosis of the liver, cystic fibrosis, cancer, Alzheimer's disease, hypertension, epilepsy, cataracts, tubulopathy, leukodystrophy, Leigh syndrome, anemia, sensorineural deafness, and hypertrophic cardiomyopathy.1-3 However, the structure of many of these proteins and the changes in their structure that lead to disease-related malfunctions are not well understood. Additionally, at least 60% of the pharmaceuticals currently available are thought to target membrane proteins, despite the fact that their exact mode of operation is not known.4-6 Developing a detailed understanding of the function of a protein is achieved by coupling biochemical experiments with knowledge of the structure of the protein. Currently the most common method for obtaining three-dimensional structure information is X-ray crystallography. However, no a priori methods are currently available to predict crystallization conditions for a given protein.7-14 This limitation is currently overcome by screening a large number of possible combinations of precipitants, buffer, salt, and pH conditions to identify conditions that are conducive to crystal nucleation and growth.7,9,11,15-24 Unfortunately, these screening efforts are often limited by difficulties associated with quantity and purity of available protein samples. While the two most significant bottlenecks for protein structure determination in general are the (i) obtaining sufficient quantities of high quality protein samples and (ii) growing high quality protein crystals that are suitable for X-ray structure determination,7,20,21,23,25-47 membrane proteins present additional challenges. For crystallization it is necessary to extract the membrane proteins from the cellular membrane. However, this process often leads to denaturation. In fact, membrane proteins have proven to be so difficult to crystallize that of the more than 66,000 structures deposited in the Protein Data Bank,48 less than 1% are for membrane proteins, with even fewer present at high resolution (< 2Å)4,6,49 and only a handful are human membrane proteins.49 A variety of strategies including detergent solubilization50-53 and the use of artificial membrane-like environments have been developed to circumvent this challenge.43,53-55 In recent years, the use of a lipidic mesophase as a medium for crystallizing membrane proteins has been demonstrated to increase success for a wide range of membrane proteins, including human receptor proteins.54,56-62 This in meso method for membrane protein crystallization, however, is still by no means routine due to challenges related to sample preparation at sub-microliter volumes and to crystal harvesting and X-ray data collection. This dissertation presents various aspects of the development of a microfluidic platform to enable high throughput in meso membrane protein crystallization at a level beyond the capabilities of current technologies. Microfluidic platforms for protein crystallization and other lab-on-a-chip applications have been well demonstrated.9,63-66 These integrated chips provide fine control over transport phenomena and the ability to perform high throughput analyses via highly integrated fluid networks. However, the development of microfluidic platforms for in meso protein crystallization required the development of strategies to cope with extremely viscous and non-Newtonian fluids. A theoretical treatment of highly viscous fluids in microfluidic devices is presented in Chapter 3, followed by the application of these strategies for the development of a microfluidic mixer capable of preparing a mesophase sample for in meso crystallization at a scale of less than 20 nL in Chapter 4. This approach was validated with the successful on chip in meso crystallization of the membrane protein bacteriorhodopsin. In summary, this is the first report of a microfluidic platform capable of performing in meso crystallization on-chip, representing a 1000x reduction in the scale at which mesophase trials can be prepared. Once protein crystals have formed, they are typically harvested from the droplet they were grown in and mounted for crystallographic analysis. Despite the high throughput automation present in nearly all other aspects of protein structure determination, the harvesting and mounting of crystals is still largely a manual process. Furthermore, during mounting the fragile protein crystals can potentially be damaged, both from physical and environmental shock. To circumvent these challenges an X-ray transparent microfluidic device architecture was developed to couple the benefits of scale, integration, and precise fluid control with the ability to perform in situ X-ray analysis (Chapter 5). This approach was validated successfully by crystallization and subsequent on-chip analysis of the soluble proteins lysozyme, thaumatin, and ribonuclease A and will be extended to microfluidic platforms for in meso membrane protein crystallization. The ability to perform in situ X-ray analysis was shown to provide extremely high quality diffraction data, in part as a result of not being affected by damage due to physical handling of the crystals. As part of the work described in this thesis, a variety of data collection strategies for in situ data analysis were also tested, including merging of small slices of data from a large number of crystals grown on a single chip, to allow for diffraction analysis at biologically relevant temperatures. While such strategies have been applied previously,57,59,61,67 they are potentially challenging when applied via traditional methods due to the need to grow and then mount a large number of crystals with minimal crystal-to-crystal variability. The integrated nature of microfluidic platforms easily enables the generation of a large number of reproducible crystallization trials. This, coupled with in situ analysis capabilities has the potential of being able to acquire high resolution structural data of proteins at biologically relevant conditions for which only small crystals, or crystals which are adversely affected by standard cryocooling techniques, could be obtained (Chapters 5 and 6). While the main focus of protein crystallography is to obtain three-dimensional protein structures, the results of typical experiments provide only a static picture of the protein. The use of polychromatic or Laue X-ray diffraction methods enables the collection of time resolved structural information. These experiments are very sensitive to crystal quality, however, and often suffer from severe radiation damage due to the intense polychromatic X-ray beams. Here, as before, the ability to perform in situ X-ray analysis on many small protein crystals within a microfluidic crystallization platform has the potential to overcome these challenges. An automated method for collecting a "single-shot" of data from a large number of crystals was developed in collaboration with the BioCARS team at the Advanced Photon Source at Argonne National Laboratory (Chapter 6). The work described in this thesis shows that, even more so than for traditional structure determination efforts, the ability to grow and analyze a large number of high quality crystals is critical to enable time resolved structural studies of novel proteins. In addition to enabling X-ray crystallography experiments, the development of X-ray transparent microfluidic platforms also has tremendous potential to answer other scientific questions, such as unraveling the mechanism of in meso crystallization. For instance, the lipidic mesophases utilized during in meso membrane protein crystallization can be characterized by small angle X-ray diffraction analysis. Coupling in situ analysis with microfluidic platforms capable of preparing these difficult mesophase samples at very small volumes has tremendous potential to enable the high throughput analysis of these systems on a scale that is not reasonably achievable using conventional sample preparation strategies (Chapter 7). In collaboration with the LS-CAT team at the Advanced Photon Source, an experimental station for small angle X-ray analysis coupled with the high quality visualization capabilities needed to target specific microfluidic samples on a highly integrated chip is under development. Characterizing the phase behavior of these mesophase systems and the effects of various additives present in crystallization trials is key for developing an understanding of how in meso crystallization occurs. A long term goal of these studies is to enable the rational design of in meso crystallization experiments so as to avoid or limit the need for high throughput screening efforts. In summary, this thesis describes the development of microfluidic platforms for protein crystallization with in situ analysis capabilities. Coupling the ability to perform in situ analysis with the small scale, fine control, and the high throughput nature of microfluidic platforms has tremendous potential to enable a new generation of crystallographic studies and facilitate the structure determination of important biological targets. The development of platforms for in meso membrane protein crystallization is particularly significant because they enable the preparation of highly viscous mixtures at a previously unachievable scale. Work in these areas is ongoing and has tremendous potential to improve not only current the methods of protein crystallization and crystallography, but also to enhance our knowledge of the structure and function of proteins which could have a significant scientific and medical impact on society as a whole. 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Vol. 2 was presented at the Ninth Pacific Science Congress, Bangkok, Thailand, Nov. 18-30, 1957.