Orientador: Sônia Maria Alves Bueno / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-18T20:07:57Z (GMT). No. of bitstreams: 1 Pavan_GiseleLuiza_M.pdf: 8113456 bytes, checksum: 17ffd6547356e32bd63a48ccb9192979 (MD5) Previous issue date: 2011 / Resumo: Imunoglobulinas são anticorpos secretados por células imunitárias em resposta a introdução de um antígeno. A imunoglobulina G (IgG) de origem humana, com alto grau de pureza, tem sido empregada como prescrição terapêutica para inúmeros casos de doenças malignas, infecciosas, auto-imunes e inflamatórias. Normalmente, a IgG é purificada através da precipitação com etanol seguida de cromatografia de afinidade com proteína A ou G imobilizada, no entanto, por apresentar alto custo, se faz necessário o desenvolvimento de técnicas de afinidade alternativas. Membranas, nas quais podem ser imobilizados os ligantes de afinidade, tornam-se uma alternativa aos géis tradicionais apresentando maior capacidade hidrodinâmica. Neste contexto, estudou-se o efeito da remoção do sal em diferentes sistemas tamponantes na purificação de IgG a partir do soro humano por cromatografia de afinidade com íons imobilizados (IMAC), para o módulo de membranas de fibras ocas de PEVA-CM-Asp- Me2+, para os íons metálicos níquel e cobalto. De acordo com eletroforeses SDS-PAGE e análises de nefelometria das frações dos picos de proteína obtidos, a melhor condição utilizada para a purificação de IgG para os adsorventes PEVA-CM-Asp-Ni2+ e PEVA-CM-Asp-Co2+, foi em presença do sistema tamponante fosfato de sódio 25 mmol L-1 e imidazol 2 mmol L-1 a pH 7,0 utilizando o aumento da concentração de imidazol como estratégia de eluição, alcançando pureza superior a 93% para ambos íons metálicos. Determinada a melhor condição de purificação, por meio das isotermas de adsorção, foi determinada a capacidade máxima de adsorção e a constante de dissociação dos complexos CM-Asp-Ni2+-IgG e CM-Asp-Co2+-IgG que, de acordo com o ajuste dos parâmetros pelo modelo de Langmuir e Langmuir-Freundlich, demonstraram boa capacidade de adsorção e constantes de dissociação características de sistemas utilizando ligantes pseudobioespecíficos. Para as filtrações em módulo de fibras ocas, construído em nosso laboratório, determinaram-se as curvas de ruptura, sendo que os resultados obtidos para as membranas finamente cortadas foram melhores que os obtidos para os de filtração em módulo de fibras ocas / Abstract: Immunoglobulins are secreted by immune cells in response to the introduction of an antigen. Immunoglobulin G (IgG) obtained from human sources with high purity has been used as therapeutic prescription for many cases of malicious, infectious, autoimmune and inflammatory diseases. Typically, IgG is purified by precipitation with ethanol followed by affinity chromatography with immobilized protein A or G, however, due to its high cost, it is necessary to develop alternative techniques based on affinity. Affinity membranes, which may be immobilized on the affinity ligands, show a higher hydrodynamic capacity than typical gels. In this context, the effect of salt was studied in different buffer systems in the purification of IgG from human serum by Immobilized Metal-ion Affinity Chromatography (IMAC) for the module PEVA-CM-Asp-Me2+ using ions Ni2+ and Co2+. According to SDS-PAGE and nephelometric analysis fractions of protein peaks obtained, the best condition used for purification of IgG to the adsorbents PEVA-CM-Asp-Ni2+ and PEVA-CM-Asp-Co2+ was in the presence of sodium phosphate buffer system 25 mmol L-1 and 2 mmol L-1 imidazole at pH 7.0 using increasing concentration of imidazole as elution strategy, achieving a purity higher than 93% for both metal ions. The maximum adsorption capacity and dissociation constant of complexes CM-Asp-Ni2+-IgG and CM-Asp-Co2+-IgG by adsorption isotherms were determined on the best condition of purification and the adjustment of parameters by the Langmuir and Langmuir-Freundlich models, it demonstrated good adsorption capacity and dissociation constants characteristics of systems using pseudobioespecific ligands. For filtration in hollow fiber module, constructed in our laboratory, breakthrough curves were determined; also for the best condition of purification, and the results for finely chopped membranes were better than those obtained for the filtration module and hollow fibers / Mestrado / Desenvolvimento de Processos Biotecnologicos / Mestre em Engenharia Química

-&gt;Context. Optical Character Recognition (OCR), having a computer recognize text from an image, is not as intuitive as human recognition. Even small (to human eyes) degradations can thwart the OCR result. The problem is that random unknown degradations are unavoidable in a real-world setting. -&gt;Objectives. The noise tolerance of Tesseract, a state-of-the-art OCR engine, is evaluated in relation to how well it handles salt and pepper noise, a type of image degradation. Noise tolerance is measured as the percentage of aberrant pixels when comparing two images (one with noise and the other without noise). -&gt;Methods. A novel systematic approach for finding the noise tolerance of an OCR engine is presented. A simulated environment is developed, where the test parameters, called test cases (font, font size, text string), can be modified. The simulation program creates a text string image (white background, black text), degrades it iteratively using salt and pepper noise, and lets Tesseract perform OCR on it, in each iteration. The iteration process is stopped when the comparison between the image text string and the OCR result of Tesseract mismatches. -&gt;Results. Simulation results are given as changed pixels percentage (noise tolerance) between the clean text string image and the text string image the degradation iteration before Tesseract OCR failed to recognize all characters in the text string image. The results include 14400 test cases: 4 fonts (Arial, Calibri, Courier and Georgia), 100 font sizes (1-100) and 36 different strings (4*100*36=14400), resulting in about 1.8 million OCR attempts performed by Tesseract. -&gt;Conclusions. The noise tolerance depended on the test parameters. Font sizes smaller than 7 were not recognized at all, even without noise applied. The font size interval 13-22 was the peak performance interval, i.e. the font size interval that had the highest noise tolerance, except for the only monospaced font tested, Courier, which had lower noise tolerance in the peak performance interval. The noise tolerance trend for the font size interval 22-100 was that the noise tolerance decreased for larger font sizes. The noise tolerance of Tesseract as a whole, given the experiment results, was circa 6.21 %, i.e. if 6.21 % of the pixel in the image has changed Tesseract can still recognize all text in the image. / <p>42</p>

>Magister Scientiae - MSc / Salt pans are a characteristic feature of many dry deserts. The microbial communities inhabiting salt pans are thought to be particularly complex and are generally dominated by halophilic microorganisms. Although saline pools are frequently found within the hyper-arid Namib Desert, the microbial communities of these saline sites have been scarcely investigated. The aim of the present study was to characterise the archaeal, bacterial and cyanobacterial diversity inhabiting these extreme saline pools using three culture independent molecular techniques (DGGE, T-RFLP and 16S rRNA clone libraries). The physiochemical results, mainly the conductivity readings recorded from the sampling sites, indicated that the Gobabeb (103.0mS/cm) region was less saline than the two Swakopmund [(Sps01) (150.0mS/cm) and Sps02 (180.0mS/cm)] sites. Results obtained from DGGE and T-RFLP data were in agreement for both bacterial and cyanobacterial analysis indicating that the Gobabeb site was more diverse than the two Swakopmund sites (Sps01 and Sps02). In comparison, the archaeal community profiles for DGGE and T-RFLP analysis were in agreement illustrating that the archaeal community were more abundant in the two extreme Swakopmund saline sites. Phylogenetic data obtained from 16S rRNA gene clone libraries identified halophilic phylotypes (Rhodothermaceae, Idiomarinaceae Puniceicoccaceae and Cyanobacteria/Chloroplast, Family VII) normally associated with salt rich sites. In addition, a large number of unclassified taxa were identified. To conclude, the study highlighted the presence of a rich microbial diversity present within the salt pans of the Namib Desert and establishes a platform for future investigations. / National Research Foundation

Managed realignment is now widely seen as an important part of coastal management strategy, as an environmentally sustainable, cost-effective alternative to traditional, hard defences. However, the trajectory of salt marsh development in managed realignment schemes remains uncertain and it is unclear how sites should be managed to fulfil both coastal defence and biodiversity objectives. In this study, the overarching aim is to find out ways in which we can create salt marshes that are self-sustaining and function as closely to natural ecosystems as possible, by considering the linked biogeomorphological processes in salt marsh ecosystems. Such an approach will improve predictions of habitat development and recommendations for future practice in managed realignment schemes. The first important question is: How do pre-alignment plant species growing within managed realignment sites respond to salt water inundation upon re-exposure to flooding? Understanding the responses of the terrestrial vegetation community to initial seawater flooding may improve predictions of the short-term transition into salt marsh vegetation. To answer this, the effects of seawater inundation on pre-existing vegetation are initially examined in a greenhouse experiment in Chapter 3. It was demonstrated that one common plant species component of many coastal grasslands, Trifolium repens, responds poorly to simulated seawater soil flooding, but the response is population-, i.e. ecotype-, specific; therefore, the species consequently has an adaptive capacity to withstand short periods of soil inundation by seawater. In addition, I look at how and why the vegetation community of a restored site transitions following the reintroduction of tidal water, including the response of the original community of non-salt marsh plant species to salt water inundation and subsequent salt marsh plant community reassembly. After three years of tidal inundation at South Efford managed realignment site (SEM), terrestrial vegetation had decreased in cover and nearly all species recorded on the adjacent natural marsh had colonised. However, the cover of salt marsh species was limited by waterlogging, caused by modifications to the tidal regime by a self-regulating tidal gate. This leads on to the second question: How do new engineering techniques alter the tidal regime and what specific aspects of the new regime drive plant community reassembly and sedimentation patterns? In Chapters 4, 5 and 6, three years of ecological and geomorphological development are investigated in response to a variable inundation regime imposed by regulated tidal exchange at SEM. Inundation of the marsh surface was very regular, but water levels were not deep enough to encourage sufficient morphological development, sedimentation nor hydrochory. In contrast, ecological development was limited by waterlogging. Balancing the tidal regime with the drainage efficiency of managed realignment sites may be the most likely scenario under which restored salt marsh will develop with maximum biodiversity benefits. Otherwise, further management techniques, such as the excavation of tidal channels, may need to be employed to improve site drainage. Consequently, the final question is: How can biodiversity be maximized on realignment sites through the use of different management techniques and site design? Tidal channels on a range of managed and natural sites were shown to improve the drainage efficiency of adjacent soils (particularly channels of greater width and/or higher Strahler order). Plant species diversity was generally higher on channel banks in managed realignment sites. On sites with highly reduced soils, the colonisation and establishment of halophytes could be advanced on the banks of tidal channels. Additionally, topographic heterogeneity introduced by tidal channels created a variety of habitat niches, which allowed a range of salt marsh species to establish in the absence of highly competitive species, such as Elymus repens. Results from this study could contribute to the generation of a number of recommendations for the implementation of managed realignment schemes, particularly regarding the excavation of tidal creek networks.

Philosophiae Doctor - PhD / This study explores the roles of exogenously applied nitric oxide, exogenously applied caffeic acid and salt stress on the antioxidant system in cereal (exemplified by maize) and legume (using soybean as an example) plants together with their influence on membrane integrity and cell death.This study investigates changes in H2O2 content, root lipid peroxidation, root cell death and antioxidant enzymatic activity in maize roots in response to exogenously applied nitric oxide (NO) and salt stress. This part of the study is based on the partially understood interaction between NO and reactive oxygen species (ROS) such as H2O2 and the role of antioxidant enzymes in plant salt stress responses. The results show that application of salt (NaCl) results in elevated levels of H2O2 and an increase in lipid peroxidation, consequently leading to increased cell death. The study also shows that by regulating the production and detoxification of ROS through modulation of antioxidant enzymatic activities, NO plays a pivotal role in maize responses to salt stress. The study argues for NO as a regulator of redox homeostasis that prevents excessive ROS accumulation during exposure of maize to salinity stress that would otherwise be deleterious to maize. This study extends the role of exogenously applied NO to improve salt stress tolerance in cereals crops (maize) further to its role in enhancing salt stress tolerance in legumes. The effect of long-term exposure of soybean to NO and salt stress on root nodule antioxidant activity was investigated to demonstrate the role of NO in salt stress tolerance. The results show that ROS scavenging antioxidative enzymes like SOD, GPX and GR are differentially regulated in response to exogenous application of NO and salt stress. It remains to be determined if the NOinduced changes in antioxidant enzyme activity under salt stress are sufficient to efficiently reduce ROS accumulation in soybean root nodules to levels close to those of unstressed soybean root nodules. Furthermore, this study investigates the effect of long-term exposure of soybean to exogenous caffeic acid (CA) and salt stress, on the basis of the established role of CA as an antioxidant and the involvement of antioxidant enzymes in plant salt stress responses. The effect of CA on soybean nodule number, biomass (determined on the basis of nodule dry weight, root dry weight and shoot dry weight), nodule NO content, and nodule cyclic guanosine monophosphate (cGMP) content in response to salt stress was investigated. Additionally, CA-induced changes in nodule ROS content, cell viability, lipid peroxidation and antioxidant enzyme activity as well as some genes that encode antioxidant enzymes were investigated in the presence or absence of salt stress. The study shows that long-term exposure of soybean to salt stress results in reduced biomass associated with accumulation of ROS, elevated levels of lipid peroxidation and elevated levels of cell death. However, exogenously applied CA reversed the negative effects of salt stress on soybean biomass, lipid peroxidation and cell death. CA reduced the salt stress-induced accumulation of ROS by mediating changes in root nodule antioxidant enzyme activity and gene expression. These CA-responsive antioxidant enzymes were found to be superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione peroxidase (GPX), and glutathione reductase (GR), which contributed to the scavenging of ROS in soybean nodules under salt stress. The work reported in Chapter 2 has been published in a peer-reviewed journal [Keyster M, Klein A, Ludidi N (2012) Caspase-like enzymatic activity and the ascorbate-glutathione cycle participate in salt stress tolerance of maize conferred by exogenously applied nitric oxide. Plant Signaling and Behavior 7: 349-360]. My contribution to the published paper was all the work that is presented in Chapter 2,whereas the rest of the work in the paper (which is not included in Chapter 2) was contributed by Dr Marshall Keyster.

>Magister Scientiae - MSc / Nitric oxide (NO) has been shown to orchestrate multiple defense responses to both abiotic and biotic stress. Importantly, elevation of nitric oxide content in plants by using nitric oxide generating compounds has been shown to enhance plant tolerance to abiotic stresses such as salt and drought via up-regulation of genes involved in the regulation of plant responses to abiotic stress. In this study, the effect(s) of nitric oxide (generated from 10 μM of the nitric oxide donor DET/NO) on the expression of a novel soybean cystatin gene (Glyma20g08800), lipid peroxidation, caspase-like activity and cell death in salt (150 mM)-stressed soybean leaves, roots and nodules were investigated. Salt treatment resulted in elevated lipid peroxidation, caspase-like activity and increased cell death in organs studied while the observed detrimental effects of salt stress were reversed by NO treatment. Salt stress suppressed the expression of Glyma20g08800 while the levels of expression of Glyma20g08800 returned towards those of unstressed plants when the salt-stressed plants were supplemented with nitric oxide (DETA/NO). Furthermore, promoter sequences of GmCYS1p626 and three of its homologues (Glyma20g08800, Glyma14g04250 and Glyma18g12240) were analyzed for putative abiotic stress and/NO cisregulatory elements based on co-expression analyses using bioinformatics. Several abiotic stress induced transcription factors (TFs) were identified and were hypothesized to be co-acting either directly or indirectly through additional factors in the regulation of soybean cystatin expression in response to NO and abiotic stress. Taken together, these results highlight the possibility of using NO to drive high levels of expression of cystatins during salt stress and lead to accumulation of the cystatin to levels that are sufficient to inhibit salt stress-induced caspase-like activity, which will inhibit salt stress-induced cell death and thus enhance the tolerance of the plant to salt stress and possibly tolerance to drought stress as well.

The primary objective of this research is to develop an applied technology and provide an assessment for remotely measuring and analyzing the real time or near real time concentrations of used nuclear fuel (UNF) elements in electrorefiners (ER). Here, Laser-Induced Breakdown Spectroscopy (LIBS) in UNF pyroprocessing facilities was investigated. LIBS is an elemental analysis method, which is based on the emission from plasma generated by focusing a laser beam into the medium. This technology has been reported to be applicable in solids, liquids (includes molten metals), and gases for detecting elements of special nuclear materials. The advantages of applying the technology for pyroprocessing facilities are: (i) Rapid real-time elemental analysis; (ii) Direct detection of elements and impurities in the system with low limits of detection (LOD); and (iii) Little to no sample preparation is required. One important challenge to overcome is achieving reproducible spectral data over time while being able to accurately quantify fission products, rare earth elements, and actinides in the molten salt. Another important challenge is related to the accessibility of molten salt, which is heated in a heavily insulated, remotely operated furnace in a high radiation environment within an argon gas atmosphere. This dissertation aims to address these challenges and approaches in the following phases with their highlighted outcomes: 1. Aerosol-LIBS system design and aqueous testing: An aerosol-LIBS system was designed around a Collison nebulizer and tested using deionized water with Ce, Gd, and Nd concentrations from 100 ppm to 10,000 ppm. The average %RSD values between the sample repetitions were 4.4% and 3.8% for the Ce and Gd lines, respectively. The univariate calibration curve for Ce using the peak intensities of the Ce 418.660 nm line was recommended and had an R2 value, LOD, and RMSECV of 0.994, 189 ppm, and 390 ppm, respectively. The recommended Gd calibration curve was generated using the peak areas of the Gd 409.861 nm line and had an R2, LOD, and RMSECV of 0.992, 316 ppm, and 421 ppm, respectively. The partial least squares (PLS) calibration curves yielded similar results with RMSECV of 406 ppm and 417 ppm for the Ce and Gd curves, respectively. 2. High temperature aerosol-LIBS system design and CeCl3 testing: The aerosol-LIBS system was transitioned to a high temperature and used to measure Ce in molten LiCl-KCl salt within a glovebox environment. The concentration range studied was from 0.1 wt% to 5 wt% Ce. Normalization was necessary due to signal degradation over time; however, with the normalization the %RSD values averaged 5% for the mid and upper concentrations studied. The best univariate calibration curve was generated using the peak areas of the Ce 418.660 nm line. The LOD for this line was 148 ppm with the RMSECV of 647 ppm. The PLS calibration curve was made using 7 latent variables (LV) and resulting in the RMSECV of 622 ppm. The LOD value was below the expected rare earth concentration within the ER. 3. Aerosol-LIBS testing using UCl3: Samples containing UCl3 with concentrations ranging from 0.3 wt% to 5 wt% were measured. The spectral response in this range was linear. The best univariate calibration curves were generated using the peak areas of the U 367.01 nm line and had an R2 value of 0.9917. Here, the LOD was 647 ppm and the RMSECV was 2,290 ppm. The PLS model was substantially better with a RMSECV of 1,110 ppm. The LOD found here is below the expected U concentrations in the ER. The successful completion of this study has demonstrated the feasibility of using an aerosol-LIBS analytical technique to measure rare earth elements and actinides in the pyroprocessing salt.

Aluminium alloys are frequently pre-treated by a conversion coating before application of an organic coating in order to improve the corrosion resistance and adhesive properties of the surface and the corrosion resistance provided by the system. Chromate-containing conversion coatings are commonly used for this purpose. However, legislation limits future use of hexavalent chromium compounds due to their toxic and carcinogenic nature. Therefore, alternative, so-called chromium-free conversion coatings are being developed that are more environmentally-compliant.The purpose of the present work has therefore been to contribute to a better understanding of how the aluminium substrate affects the formation and properties of conversion coatings for adhesive bonding. In particular, a chrome-free zirconium-based conversion treatment process has been investigated as a possible replacement for conventional chromate conversion treatment. The influence of the conversion time on the thickness of the formed layer on pure aluminium was investigated using complementary surface analytical techniques. The conversion time was varied between 30 and 600 seconds.In this study, the structure and composition of zirconium-based chromium-free conversion coatings on magnetron sputtered superpure aluminium and a range of aluminium-copper alloys were characterised as a function of immersion time in the aqueous conversion bath to understand the mechanism of coating formation and protection. However, the presence of copper significantly influences the coating development and ultimately the performance of the conversion coatings formed on binary copper-containing aluminium alloys.The morphology and composition of the coatings have been probed using transmission electron microscopy, Rutherford backscattering spectroscopy and glow discharge optical emission spectroscopy, with loss of substrate through growth of the conversion coating also quantified. A comparison of the RBS spectra obtained for the superpure aluminium specimens after different immersion times revealed that zirconium (Zr) and oxygen (O) peaks were wider for longer immersion times, indicating thickening of the coating with increased immersion times. Thus, increasing the immersion time resulted in an increase in coating thickness but little change in coating composition occurred as determined by the RBS RUMP simulations. Alloying decreases the coating thickness, as well as metal consumption. Here, aspects of the corrosion behaviour of superpure aluminium and aluminium-copper alloys were also considered using electronoptical, electrochemical and surface analytical probing. The influence that short and prolonged treatment times exert on the performances of such conversion coating is discussed. The conversion coating formed after 60 s and 180 s of immersion in the zirconium-based conversion coating bath provide good corrosion resistance which can be attributed to the high stability of the compounds that constitute the surface oxide layer, and good adhesion properties.

No description available.

There is a paucity of published information on fish utilization of salt marshes outside North America. This dissertation represents the first intensive examination of the ichthyofauna associated with salt marshes in southern Africa and examines the species composition, diversity, size structure, distribution and estuarine dependence of fishes that utilize salt marshes in the Kariega Estuary. The research was concentrated on Taylor's salt marsh, with comparative studies being conducted on other salt marshes and habitats within the marine dominated Kariega Estuary. The possible importance of salt marshes as feeding and refuge areas for fishes was examined, as was the role of salt marshes in the food web of the estuary. The fishes frequenting salt marshes in the Kariega Estuary were predominantly the juveniles of marine species, with Mugilidae being the dominant family. The ichthyofauna was distributed primarily in the intertidal creeks with very few specimens captured on the vegetated Spartina maritima and Sarcocomia perennis flats. The different reaches of the intertidal creek were characterised by distinct fish assemblages. The fish assemblages associated with the intertidal salt marsh creeks were significantly different from those found in the eelgrass beds, the other dominant intertidal habitat in the Kariega Estuary. The eelgrass beds were dominated by estuarine fish species and had a higher density and standing stock of fishes when compared to the salt marsh creeks. The diversity of fishes in the two habitats was however similar. The intertidal salt marsh creek ichthyofauna also differed significantly from that found in the main estuary channel. The creek ichthyofauna was dominated by 0+ juveniles while the main channel had many subadult and adult fishes. The channel habitat also had numerous large piscivorous fishes which were absent from the salt marsh creeks. The low number of piscivorous fishes, together with limited fish predation from other sources, may be the reason why salt marshes provide a refuge for juvenile fishes that frequent these habitats. Unlike previous studies on North American, Australian and European salt marshes, the dominant fish species that frequented Taylor's marsh were not recorded feeding extensively on the marsh, and those that did had a limited distribution. The role of fishes in the transfer of energy off the Kariega salt marshes is therefore likely to be minimal. A stable carbon isotope study on the dominant primary producers and consumers within the Kariega Estuary revealed that detritus originating from the high lying salt marsh plants Sarcocornia perennis and Chenolea diffusa was not utilized by fishes in the Kariega Estuary. Preliminary results indicated that the cord grass Spartina maritima may be an important energy source to the fishes in the Kariega Estuary, but further research is needed to confirm this.