Comparison between hydrographically conditioned digital elevation models in the morphometric charaterization of watersheds

The aim of this study was to compare the hydrographically conditioned digital elevation models (HCDEMs) generated from data of VNIR (Visible Near Infrared) sensor of ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer), of SRTM (Shuttle Radar Topography Mission) and topographical maps from IBGE in a scale of 1:50,000, processed in the Geographical Information System (GIS), aiming the morphometric characterization of watersheds. It was taken as basis the Sub-basin of São Bartolomeu River, obtaining morphometric characteristics from HCDEMs. Root Mean Square Error (RMSE) and cross validation were the statistics indexes used to evaluate the quality of HCDEMs. The percentage differences in the morphometric parameters obtained from these three different data sets were less than 10%, except for the mean slope (21%). In general, it was observed a good agreement between HCDEMs generated from remote sensing data and IBGE maps. The result of HCDEM ASTER was slightly higher than that from HCDEM SRTM. The HCDEM ASTER was more accurate than the HCDEM SRTM in basins with high altitudes and rugged terrain, by presenting frequency altimetry nearest to HCDEM IBGE, considered standard in this study.

A Farewell to Flat Biology. Three-dimensional Cell Culture Models in Cancer Drug Target Identification and Validation

Cells of epithelial origin, e.g. from breast and prostate cancers, effectively differentiate into complex multicellular structures when cultured in three-dimensions (3D) instead of conventional two-dimensional (2D) adherent surfaces. The spectrum of different organotypic morphologies is highly dependent on the culture environment that can be either non-adherent or scaffold-based. When embedded in physiological extracellular matrices (ECMs), such as laminin-rich basement membrane extracts, normal epithelial cells differentiate into acinar spheroids reminiscent of glandular ductal structures. Transformed cancer cells, in contrast, typically fail to undergo acinar morphogenic patterns, forming poorly differentiated or invasive multicellular structures. The 3D cancer spheroids are widely accepted to better recapitulate various tumorigenic processes and drug responses. So far, however, 3D models have been employed predominantly in the Academia, whereas the pharmaceutical industry has yet to adopt a more widely and routine use. This is mainly due to poor characterisation of cell models, lack of standardised workflows and high throughput cell culture platforms, and the availability of proper readout and quantification tools. In this thesis, a complete workflow has been established entailing well-characterised 3D cell culture models for prostate cancer, a standardised 3D cell culture routine based on high-throughput-ready platform, automated image acquisition with concomitant morphometric image analysis, and data visualisation, in order to enable large-scale high-content screens. Our integrated suite of software and statistical analysis tools were optimised and validated using a comprehensive panel of prostate cancer cell lines and 3D models. The tools quantify multiple key cancer-relevant morphological features, ranging from cancer cell invasion through multicellular differentiation to growth, and detect dynamic changes both in morphology and function, such as cell death and apoptosis, in response to experimental perturbations including RNA interference and small molecule inhibitors. Our panel of cell lines included many non-transformed and most currently available classic prostate cancer cell lines, which were characterised for their morphogenetic properties in 3D laminin-rich ECM. The phenotypes and gene expression profiles were evaluated concerning their relevance for pre-clinical drug discovery, disease modelling and basic research. In addition, a spontaneous model for invasive transformation was discovered, displaying a highdegree of epithelial plasticity. This plasticity is mediated by an abundant bioactive serum lipid, lysophosphatidic acid (LPA), and its receptor LPAR1. The invasive transformation was caused by abrupt cytoskeletal rearrangement through impaired G protein alpha 12/13 and RhoA/ROCK, and mediated by upregulated adenylyl cyclase/cyclic AMP (cAMP)/protein kinase A, and Rac/ PAK pathways. The spontaneous invasion model tangibly exemplifies the biological relevance of organotypic cell culture models. Overall, this thesis work underlines the power of novel morphometric screening tools in drug discovery.

Comparison of full-field electroretinogram in diabetic and non diabetic dogs with cataracts

Being the commonest ocular disorder, dense cataracts disable fundoscopic examination and the diagnosis of retinal disorders, which dogs may be predisposed. The aim of this study was to compare the electroretinographic responses recorded according to the International Society for Clinical Electrophysiology of Vision human protocol to evaluate retinal function of diabetic and non diabetic dogs, both presenting mature or hypermature cataracts. Full-field electroretinogram was recorded from 66 dogs, with ages varying from 6 to 15 years old allocated into two groups: (1) CG, non diabetic cataractous dogs, and (2) DG, diabetic cataractous dogs. Mean peak-to-peak amplitude (microvolts) and b-wave implicit time (milliseconds) were determined for each of the five standard full-field ERG responses (rod response, maximal response, oscillatory potentials, single-flash cone response and 30 Hz flicker). Comparing CG to DG, ERGs recorded from diabetic dogs presented lower amplitude and prolonged b-wave implicit time in all ERG responses. Prolonged b-wave implicit time was statistically significant (p< 0.05) at 30 Hz flicker (24.0 ms versus 22.4 ms). These data suggests full-field ERG is capable to record sensible alterations, such as flicker's implicit time, being useful to investigate retinal dysfunction in diabetic dogs.

Normal values of b-wave responses of full-field electroretinogram in Lhasa Apso dogs with cataracts according to age

Lhasa Apso dogs with immature, mature or hypermature cataracts were divided into four groups according to their age (G1: 1 to 3 years old, G2: 4 to 7 years old, G3: 8 to 11 years old, G4: more than 12 years old). All animals were evaluated under the same sedation protocol to allow the performance of the electroretinogram (ERG) exam to determine normal value of b-wave response of the full-field ERG according to age. Three ERG responses were recorded: rod, maximal and cone responses. The amplitude values and b-wave implicit time of the responses of all groups were compared and analyzed by Kruskal-Wallis test (variance analysis for non-repeated measures), followed by the Dunn post-test (when p<0,05). A significant decrease was observed in maximal responses' amplitude, when comparing the G4 group with G1 and G2. No statistically relevant differences were observed in the b-wave implicit time values between groups. The ERG values are directly influenced by the animal's age. Older patients presented a decrease in the amplitude of the maximal response. The study determined the normal parameters of ERG b-waves for Lhasa Apso dogs with cataract according to their age group.

Globe fringerush (Fimbristylis miliacea) cross resistance to als-inhibitor herbicides under field conditions in irrigated rice in the south of Brazil

ALS-inhibiting herbicides usually provide adequate weed control in irrigated rice fields. After consecutive years of use, the Cyperaceae species, globe fringerush (Fimbristylis miliacea) began to show resistance to ALS (acetolactate synthase) inhibitors. Globe fringerush is one of the most problematic herbicide-resistant weeds in irrigated rice in the state of Santa Catarina in the South of Brazil. The objective of this research was to examine cross resistance of globe fringerush to ALS inhibitors, under field conditions. Two experiments were conducted in a rice field naturally infested with ALS-resistant globe fringerush in Santa Catarina, in the 2008/09 and 2009/10 cropping seasons. The experimental units were arranged in randomized complete block design, with five replicates, consisting of two factors (herbicide and dose) in a 4 x 5 factorial arrangement. ALS herbicides included bispyribac-sodium, ethoxysulfuron, pyrazosulfuron-ethyl and penoxsulam. Six-leaf globe fringerush was sprayed with herbicide doses of 0, 0.5, 1, 2 and 4X the recommended doses in a spray volume of 200 L ha-1. The number of rice culm, filled and sterile grains, plant height, dry shoot biomass and grain yield were recorded. Globe fringerush control was evaluated 28 and 70 days after herbicide application (DAA); shoots were harvested at 13 weeks after herbicide application and dry weight recorded. Competition with globe fringerush reduced the number of culm and rice grain yield. The globe fringerush biotype in this field was resistant to all ALS herbicides tested. Penoxsulam had the highest level of activity among treatments at 28 and 70 DAA, but the control level was only 50% and 42%, respectively, in the second year of assessment. This was not enough to prevent rice yield loss. Alternative herbicides and weed control strategies are necessary to avoid yield losses in rice fields infested with ALS-resistant biotypes of globe fringerush.

The potential of natural regeneration of rocky outcrop vegetation on rupestrian field soils in "Serra do Cipó", Brazil

An area's innate potential to regenerate represents a crucial factor for its conservation and management. The seed rain and seed bank are important agents in the regeneration process. Seed banks are particularly important in communities where there is a high proportion of obligate seeders. Rocky outcrops are habitats where most part of the plant species depends on their seeds to reproduce and maintain viable populations. Therefore, seed banks ought to be important in this vegetation physiognomy. We test the hypotheses that the seed bank of the rocky outcrops found in the rupestrian fields of "Serra do Cipó", Brazil, is richer in species and denser than those formed on different vegetation physiognomies neighboring the outcrops. We then compared species abundance, species richness and composition in the rocky outcrops' seed banks with those of sandy and peaty bogs, forests, gallery forests, and "cerrados". Furthermore, we report on the natural regeneration potential of these soils by assessing a greenhouse study on seedling emergence. Soil samples were collected from 0 to 5 and 5 to 10 cm of depth. Rocky outcrops had the poorest in species and less dense seed bank and showed segregation in species composition. Emergence was greater in the most superficial layer. However, soils on rocky outcrops showed the greatest proportion of endemic threatened species in their seed banks, demonstrating their importance for biodiversity conservation of the "Serra do Cipó" rupestrian fields.

Evidencing and Sharing Value with Decision Making Models in an Industrial Maintenance Network

The objective of this thesis is to concretize the potential benefits that the industrial maintenance case network could achieve through using the value-based life-cycle model and the flexible asset management model. It is also inspected what factors prevent value creation and sharing in the maintenance contract practices of the case network. This thesis is a case study which utilizes modelling. Four scenarios were developed to demonstrate value creation in the future. The data was partly provided by the collaborating company, partly gathered from public financial statement information. The results indicate that value has been created through the past maintenance of the collaborating company’s rod mill and that profitability of the collaborating company has been mostly on satisfactory level during the past few years. Potential value might be created by increasing the share of proactive maintenance of the rod mill in the future. Profitability of the network could be improved in the future through flexible asset management operations. The main obstacle for value creation and sharing seems to be the lack of sufficient trust between the network members.

Future of Newspaper Business Models in Finland

The purpose of this Master’s thesis was to study the business model development in Finnish newspaper industry during the next then years through scenario planning. The objective was to see how will the business models develop amidst the many changes in the industry, what factors are affecting the change, what are the implications of these changes for the players in the industry and how should the Finnish newspaper companies evolve in order to succeed in the future. In this thesis the business model change is studied based on all the elements of business models, as it was discovered that the industry is too often focusing on changes in only few of those elements and a more broader view can provide valuable information for the companies. The results revealed that the industry is affected by many changes during the next ten years. Scenario planning provides a good tool for analyzing this change and for developing valuable options for businesses. After conducting series of interviews and discovering forces affecting the change, four different scenarios were developed centered on the role that newspaper will take and the level at which they are providing the content in the future. These scenarios indicated that there are varieties of options in the way the business models may develop and that companies should start making decisions proactively in order to succeed. As the business model elements are interdepended, changes made in the other elements will affect the whole model, making these decisions about the role and level of content important for the companies. In the future, it is likely that the Finnish newspaper industry will include many different kinds of business models, some of which can be drastically different from the current ones and some of which can still be similar, but take better into account the new kind of media environment.

Effect of race, genetic population structure, and genetic models in two-locus association studies: clustering of functional renin-angiotensin system gene variants in hypertension association studies

Previous genetic association studies have overlooked the potential for biased results when analyzing different population structures in ethnically diverse populations. The purpose of the present study was to quantify this bias in two-locus association studies conducted on an admixtured urban population. We studied the genetic structure distribution of angiotensin-converting enzyme insertion/deletion (ACE I/D) and angiotensinogen methionine/threonine (M/T) polymorphisms in 382 subjects from three subgroups in a highly admixtured urban population. Group I included 150 white subjects; group II, 142 mulatto subjects, and group III, 90 black subjects. We conducted sample size simulation studies using these data in different genetic models of gene action and interaction and used genetic distance calculation algorithms to help determine the population structure for the studied loci. Our results showed a statistically different population structure distribution of both ACE I/D (P = 0.02, OR = 1.56, 95% CI = 1.05-2.33 for the D allele, white versus black subgroup) and angiotensinogen M/T polymorphism (P = 0.007, OR = 1.71, 95% CI = 1.14-2.58 for the T allele, white versus black subgroup). Different sample sizes are predicted to be determinant of the power to detect a given genotypic association with a particular phenotype when conducting two-locus association studies in admixtured populations. In addition, the postulated genetic model is also a major determinant of the power to detect any association in a given sample size. The present simulation study helped to demonstrate the complex interrelation among ethnicity, power of the association, and the postulated genetic model of action of a particular allele in the context of clustering studies. This information is essential for the correct planning and interpretation of future association studies conducted on this population.

Influence of Pulsed-Electric Field Irradiation in Crystallization

The growing pharmaceutical interest, among others, in the polymorphic composition of the emerging solid end-products from production processes has been traced to the need for attainment of high product purity. This is more so as the presence of different polymorphs may constitute physical impurity of the product. Hence, the need for optimization of the yield of desired product component(s) through controlled crystallization kinetics for instance. This study was carried out to investigate the impact of pulsed electric field (PEF) irradiation on the crystal morphology of glycine obtained by cooling crystallization (without seeding) from commercial glycine sample in distilled deionized water solution. In doing so, three different pulse frequencies (294, 950 and 145 Hz) and a case without PEF were studied at three cooling rates (5, 10 and 20 ºC/h). The crystal products obtained were analyzed for polymorphic composition by powder x-ray diffraction (PXRD) and Fourier transform infrared (FTIR) spectroscopy while the particles characterization was done on Morphologi G3. The results obtained from this study showed that pulsed electric field irradiation had significant impact on metastability of the aqueous solution as well as on the polymorphic composition of the end product. With increasing PEF frequency applied, nucleation started earlier and the γ-glycine polymorph content of the product crystals increased. These were found to have been aided by cooling rate, as the most significant effect was observed at 5 ºC/h. It was also discovered that PEF application had no measurable impact on the pH of the aqueous solution as well as the size distribution of the particles. Cooling on the contrary was believed to be responsible for the broadening of the particle size distribution with a downward shift of the lower limit of the raw material from about 100 μm to between 10 and 50 μm.

Three-dimensional conformal breast irradiation in the prone position

The prone position can be used for the planning of adjuvant radiotherapy after conservative breast surgery in order to deliver less irradiation to lung and cardiac tissue. In the present study, we compared the results of three-dimensional conformal radiotherapy planning for five patients irradiated in the supine and prone position. Tumor stage was T1N0M0 in four patients and T1N1M0 in one. All patients had been previously submitted to conservative breast surgery. Breast size was large in three patients and moderate in the other two. Irradiation in the prone position was performed using an immobilization foam pad with a hole cut into it to accommodate the breast so that it would hang down away from the chest wall. Dose-volume histograms showed that mean irradiation doses reaching the ipsilateral lung were 8.3 ± 3.6 Gy with the patient in the supine position and 1.4 ± 1.0 Gy with the patient in the prone position (P = 0.043). The values for the contralateral lung were 1.3 ± 0.7 and 0.3 ± 0.1 Gy (P = 0.043) and the values for cardiac tissue were 4.6 ± 1.6 and 3.0 ± 1.7 Gy (P = 0.079), respectively. Thus, the dose-volume histograms demonstrated that lung tissue irradiation was significantly lower with the patient in the prone position than in the supine position. Large-breasted women appeared to benefit most from irradiation in the prone position. Prone position breast irradiation appears to be a simple and effective alternative to the conventional supine position for patients with large breasts, since they are subjected to lower pulmonary doses which may cause less pulmonary side effects in the future.

Experimental models in vaccine research: malaria and leishmaniasis

Animal models have a long history of being useful tools, not only to test and select vaccines, but also to help understand the elaborate details of the immune response that follows infection. Different models have been extensively used to investigate putative immunological correlates of protection against parasitic diseases that are important to reach a successful vaccine. The greatest challenge has been the improvement and adaptation of these models to reflect the reality of human disease and the screening of vaccine candidates capable of overcoming the challenge of natural transmission. This review will discuss the advantages and challenges of using experimental animal models for vaccine development and how the knowledge achieved can be extrapolated to human disease by looking into two important parasitic diseases: malaria and leishmaniasis.

Air blast freezing of fruit pulp models in commercial boxes: influence of preferential channels in the bed on freezing times estimating

The freezing times of fruit pulp models packed and conditioned in multi-layered boxes were evaluated under conditions similar to those employed commercially. Estimating the freezing time is a difficult practice due to the presence of significant voids in the boxes, whose influence may be analyzed by means of various methods. In this study, a procedure for estimating freezing time by using the models described in the literature was compared with experimental measurements by collecting time/temperature data. The following results show that the airflow through packages is a significant parameter for freezing time estimation. When the presence of preferential channels was considered, the predicted freezing time in the models could be 10% lower than the experimental values, depending on the method. The isotherms traced as a function of the location of the samples inside the boxes showed the displacement of the thermal center in relation to the geometric center of the product.