Planning and ramping up a new SW Testing site

During recent years, mobile phone markets have changed significantly. Asian markets have become vital for the manufacturers with their millions of end users and multiple major mobile network operators. This has resulted in software development as global companies have research and development sites running in multiple locations, including Asia. The reasons behind this are not only in reducing labor costs but also in capitalizing on the local knowledge and knowhow. A ramp-up site has multiple effects in the software development and software release activities. This thesis focuses on representing the importance of software testing as part of software development process and highlighting issues that need to be considered during ramp-up activities. In addition this work tries to emphasize the importance of communication between parties and information gathering prior to setting up the ramp-up site. The output of this thesis was successful software testing site ramp-up within the set time limits. The quality of software testing work was assured and the ramp-up -project requirements were achieved.

The Contribution of Mobile Device Manufacturers’ Site Usage to Brand Satisfaction and Loyalty

Recently most of the mobile phone manufacturer companies started to pay extra attention to their websites. This research investigates the sources of value creation on the mobile phone manufacturers’ website and the affect visiting the manufacturers’ website has on brand loyalty and brand satisfaction. The results show a correlation between positive website usage experience, brand loyalty and brand satisfaction. Moreover there is a relation between Novelty, Efficiency, Lock-in and the perceived usefulness the manufacturers’ website has on the mobile phone device. And finally the main reason behind Finnish student’s mobile phone brand loyalty is its country of origin.

Membrane emulsification in preparation of single site catalyst

In the theory part the membrane emulsification was studied. Emulsions are used in many industrial areas. Traditionally emulsions are prepared by using high shear in rotor-stator systems or in high pressure homogenizer systems. In membrane emulsification two immiscible liquids are mixed by pressuring one liquid through the membrane into the other liquid. With this technique energy could be saved, more homogeneous droplets could be formed and the amount of surfactant could be decreased. Ziegler-Natta and single-site catalysts are used in olefin polymerization processes. Nowadays, these catalysts are prepared according to traditional mixing emulsification. More homogeneous catalyst particles that have narrower particle size distribution might be prepared with membrane emulsification. The aim of the experimental part was to examine the possibility to prepare single site polypropylene catalyst using membrane emulsification technique. Different membrane materials and solidification techniques of the emulsion were examined. Also the toluene-PFC phase diagram was successfully measured during this thesis work. This phase diagram was used for process optimization. The polytetrafluoroethylene membranes had the largest contact angles with toluene and also the biggest difference between the contact angles measured with PFC and toluene. Despite of the contact angle measurement results no significant difference was noticed between particles prepared using PTFE membrane or metal sinter. The particle size distributions of catalyst prepared in these tests were quite wide. This would probably be fixed by using a membrane with a more homogeneous pore size distribution. It is also possible that the solidification rate has an effect on the particle sizes and particle morphology. When polymeric membranes are compared PTFE is probably still the best material for the process as it had the best chemical durability.

Control of Asian soybean rust with mancozeb, a multi-site fungicide

An experiment conducted in the field the action of mancozeb, a fungicide of multi-site action was tested, to control soybean rust caused by Phakopsora pachyrhizi. Its performance was compared to that of the mixture cyproconazole (DMI) + azoxystrobin (QoI). The soybean cultivar NA 7337 RR was used with a population of 400,000 plants/ha cultivated in 20m2 plots. Treatments consisted of mancozeb levels (1.5 and 2.0 kg/ha) applied four, six and eight times. The DMI + QoI mixture was applied three times at 0.3 L/ha + Nimbus. Rust severity was assessed six times in the plots and data were integrated as the area under the disease progress curve (AUDPC). The plots were harvested and grain yield was expressed as kg/ha. Data on AUDPC and yield were subjected to analysis of variance and means compared according to Turkey's test (p = 0.005). Treatments with mancozeb were superior to DMI + QoI mixture both for rust control and grain yield. Four applications of 2.0 k/ha mancozeb were more efficient than three applications of the mixture used as standard. Mancozeb has the potential to be added to fungicide mixtures in the establishment of soybean rust anti-resistance strategy.

Disseminação seletiva da informação na área da saúde: o caso do web site Amedeo

As atividades de produção/divulgação de informações foram otimizadas pela facilidade de comunicação oferecida pela internet, e a produção científica cresce cada vez mais. Com foco nas facilidades de acesso à informação científica oferecidas pela tecnologia, o objetivo é evidenciar os benefícios da aplicação da Disseminação Seletiva da Informação (SDI) na formação de profissionais da saúde, a partir da análise do web site Amedeo. Faz-se um estudo comparativo, de modo a verificar como os artigos disseminados pelo Amedeo podem ser acessados no próprio e numa biblioteca universitária - no caso, analisou-se o acesso através do site do Sistema de Bibliotecas da Unicamp e, ainda, recorre-se à literatura para identificar relações entre a Ciência da Informação e a área de Ciências da Saúde, principalmente quanto à prática da Medicina Baseada em Evidências. Apresentam-se sugestões aos profissionais da informação e professores interessados em identificar possibilidades de uso dos recursos do Amedeo e demais serviços de SDI, em bibliotecas universitárias, de modo a colaborar com o processo educacional dos profissionais da área da saúde.

Contexts, non-specificity, and minimalism

Atlas (2007) argues that semantic minimalism (as defended by Cappelen & Lepore 2005) fails because it cannot deal with semantic non-specificity. I argue that thereis a plausible version of minimalism-viz., situated minimalism-which doesn't succumb to the non-specificity charge insofar as non-specificity can be dealt with at a postsemantic level. Thus, pragmatics plays no rolewhen it comes to determining the (minimal) proposition expressed. Instead, pragmatic and other extra-semantic considerations enter the scene in characterizing the situation vis-à-vis which the proposition is evaluated. For this reason a plausible form of minimalism must embrace a form of truth-relativism: a proposition is not universally true/false, but true/false only relative to a situation. I show how the position defended is not only (i) more cognitively plausible than either (semantic) minimalism as proposed by Cappelen & Lepore or the positions appealing to pragmaticintrusion into the proposition expressed, but is also (ii) in accordance with ordinary people's intuitions.

Dry biomass distribution in a cerrado sensu stricto site in Brazil central

The Cerrado has been the main source of firewood and charcoal in Brazil, but despite being one of the hot spots for conservation of the world's biodiversity, neither plantations of native species nor sustainable management has been adopted in the region. The aim of this work was to investigate the biomass distribution and the potential for energy production of the cerrado species. The study was conducted in a cerrado sensu stricto site at the Água Limpa Farm (15º 56'14'' S and 47º 46'08'' W) in the Cerrado Biosphere Reserve. An area of 63.54ha was divided in 20 x 50m plots and, a random sample consisting of ten of these plots, representing 1.56% of the study-site, was assessed. All woody individuals from 5 cm diameter at 30 cm above ground level were identified and measured. Each individual was felled, the twigs thinner than 3cm were discarded while the larger branches and the trunks, both with bark, were weighted separately. After that, 2.5cm transverse sections of the trunk with bark were taken at 0, 25, 50, 75 and 100% of the length. A similar sample was also taken at the base of each branch. A total of 47 species in 35 genera and 24 families were found, with an average density of 673 individuals per ha. The diameter distribution showed a reversed-J shape with 67% of the individuals up to 13cm, while the maximum diameter was 32.30cm. Seven species represented 72% of the total biomass. In general, the species with higher production per tree were among those with higher production per ha. This content was distributed by diameter classes, reaching a maximum of 2.5ton/ha between 9 to 13cm and then, decreasing to 0.96 ton/ha between 29 to 33cm diameter. Carbon sequestering was 6.2ton/ha (until the actual stage of cerrado) based on an average 50% carbon content in the dry matter. The heat combustion of the wood varied from 18,903kj/kg to 20,888kj/kg with an average of 19,942kj/kg. The smaller diameter classes fix more carbon due to the large number of small plants per ha. But, for a species that reached larger dimensions and contained individuals in all diameter classes, Vochysia thyrsoidea, one can verify an increase in carbon fixation from 1.41 kg/ha in the first class (5 to 9cm) to 138,3kg/ha in the last (25 to 33cm). That indicates that it is possible to select species that reach larger size with a higher capacity of carbon accumulation per plant. The species that reached larger dimensions, with a production per tree above average and had high calorific power values were Dalbergia miscolobium, Pterodon pubescens and Sclerolobium paniculatum. These species have potential for use in fuelwood plantations and sustainable management.

DPS-Like Peroxide Resistance Protein: Structural and Functional Studies on a Versatile Nanocontainer

Oxidative stress is a constant threat to almost all organisms. It damages a number of biomolecules and leads to the disruption of many crucial cellular functions. It is caused by reactive oxygen species (ROS), such as hydrogen peroxide (H2O₂), superoxide (•O₂ -), and hydroxyl radical (•OH). The most harmful of these compounds is •OH, which is only formed in cells in the presence of redox-cycling transition metals, such as iron and copper. Bacteria have developed a number of mechanisms to cope with ROS. One of the most widespread means employed by bacteria is the DNA-binding proteins from starved cells (Dps). Dps proteins protect the cells by binding and oxidizing Fe2+, thus greatly reducing the production of •OH. The oxidized iron is stored inside the protein as an iron core. In addition, Dps proteins bind directly to DNA forming a protective coating that shields DNA from harmful agents. Moreover, Dps proteins have been found to elicit other protective functions in cells and to participate in bacterial virulence. Dps proteins are of special importance to Streptococci owing to the lack of catalase in this genus of bacteria.This study was focused on structural and functional characterization of streptococcal Dpslike peroxide resistance (Dpr) proteins. Initially, crystal structures of Streptococcus pyogenes Dpr were determined. The data confirmed the presence of a di-metal ferroxidase center (FOC) in Dpr proteins and revealed the presence of a novel N-terminal helix as well as a surface metal-binding site. The crystal structures of Streptococcus suis Dpr complexed with transition metals demonstrated the metal specificity of the FOC. Solution binding studies also indicated the presence of a di-metal FOC. These results suggested a possible role for Dpr in the detoxification of various metals. Iron was found to mineralize inside the protein as ferrihydrite based on X-ray absorption spectroscopy data. The iron core was found to exhibit clear superparamagnetic behaviour using magnetic and Mössbauer measurements. The results from this study are expected to further increase our understanding on the binding, oxidation, and mineralization of iron and other metals in Dpr proteins. In particular, the structural and magnetic properties of the iron core can form a basis for potential new applications in nanotechnology. From the streptococcal viewpoint, the results would help in understanding better the complicated picture of bacterial pathogenesis. Dpr proteins may also provide a novel target for drug design due to their tight involvement in bacterial virulence.

JNK, a versatile regulator of kinesin-1 transport and cytoskeleton dynamics

C-Jun N-terminal kinase (JNK) is traditionally recognized as a crucial factor in stress response and inducer of apoptosis upon various stimulations. Three isoforms build the JNK subfamily of MAPK; generally expressed JNK1 and JNK2 and brain specific JNK3. Degenerative potency placed JNK in the spotlight as potential pharmacological option for intervention. Unfortunately, adverse effects of potential drugs and observation that expression of only JNK2 and JNK3 are induced upon stress, restrained initial enthusiasm. Notably, JNK1 demonstrated atypical high constitutive activity in neurons that is not responsive to cellular stresses and indicated existence of physiological activity. This thesis aimed at revealing the physiological functions of JNK1 in actin homeostasis through novel effector MARCKS-Like 1 (MARCKSL1) protein, neuronal trafficking mediated by major kinesin-1 motor protein and microtubule (MT) dynamics via STMN2/SCG10. The screen for novel physiological JNK substrates revealed specific phosphorylation of C-terminal end of MARCKSL1 at S120, T148 and T183 both ex vivo and in vitro. By utilizing site-specific mutagenesis, various actin dynamics and migrations assays we were able to demonstrate that JNK1 phosphorylation specifically facilitates F-actin bundling and thus filament stabilisation. Consecutively, this molecular mechanism was proved to enhance formation of filopodia; cell surface projections that allow cell sensing surrounding environment and migrate efficiently. Our results visualize JNK dependent and MARCKSL1 executed induction of filopodia in neurons and fibroblast indicating general mechanism. Subsequently, inactivation of JNK action on MARCKSL1 shifts cellular actin machinery into lamellipodial dynamic arrangement. Tuning of actin cytoskeleton inevitably melds with cell migration. We observed that both active JNK and JNK pseudo-phosphorylated form of MARCKSL1 reduce actin turnover in intact cells leading to overall diminished cell motility. We demonstrate that tumour transformed cells from breast, prostate, lung and muscle-derived cancers upregulate MARCKSL1. We showed on the example of prostate cancer PC-3 cell line that JNK phosphorylation negatively controls MARCKSL1 ability to induce migration, which precedes cancer cell metastasis. The second round of identification of JNK physiological substrates resulted in detection of predominant motor protein kinesin-1 (Kif5). Mass spectrometry detailed analysis showed evident endogenous phosphorylation of kinesin-1 on S176 within motor domain that interacts with MT. In vitro phosphorylation of bacterially expressed kinesin heavy chain by JNK isoforms displayed higher specificity of JNK1 when compared to JNK3. Since, JNK1 is constitutively active in neurons it signified physiological aspect of kinesin-1 regulation. Subsequent biochemical examination revealed that kinesin-1, when not phosphorylated on JNK site, exhibits much higher affinity toward MTs. Expression of the JNK non-phosphorable kinesin-1 mutant in intact cells as well as in vitro single molecule imaging using total internal reflection fluorescence microscopy indicated that the mutant loses normal speed and is not able to move processively into proper cellular compartments. We identify novel kinesin-1 cargo protein STMN2/SCG10, which along with known kinesin-1 cargo BDNF is showing impaired trafficking when JNK activity is inhibited. Our data postulates that constitutive JNK activity in neurons is crucial for unperturbed physiologically relevant transport of kinesin-1 dependant cargo. Additionally, my work helps to validate another novel physiological JNK1 effector STMN2/SCG10 as determinant of axodendritic neurites dynamics in the developing brain through regulation of MT turnover. We show successively that this increased MT dynamics is crucial during developmental radial migration when brain layering occurs. Successively, we are able to show that introduction of JNK phosphorylation mimicking STMN2/SCG10 S62/73D mutant rescues completely JNK1 genetic deletion migration phenotype. We prove that STMN2/SCG10 is predominant JNK effector responsible for MT depolymerising activity and neurite length during brain development. Summarizing, this work describes identification of three novel JNK substrates MARCKSL1, kinesin-1 and STMN2/SCG10 and investigation of their roles in cytoskeleton dynamics and cargo transport. This data is of high importance to understand physiological meaning of JNK activity, which might have an adverse effect during pharmaceutical intervention aiming at blocking pathological JNK action.

Prediction of 3D structure and ligand-binding properties : ABC transporters and flavodiiron proteins from Synechocystis sp. strain PCC 6803

Cyanobacteria are unicellular, non-nitrogen-fixing prokaryotes, which perform photosynthesis similarly as higher plants. The cyanobacterium Synechocystis sp. strain PCC 6803 is used as a model organism in photosynthesis research. My research described herein aims at understanding the function of the photosynthetic machinery and how it responds to changes in the environment. Detailed knowledge of the regulation of photosynthesis in cyanobacteria can be utilized for biotechnological purposes, for example in the harnessing of solar energy for biofuel production. In photosynthesis, iron participates in electron transfer. Here, we focused on iron transport in Synechocystis sp. strain PCC 6803 and particularly on the environmental regulation of the genes encoding the FutA2BC ferric iron transporter, which belongs to the ABC transporter family. A homology model built for the ATP-binding subunit FutC indicates that it has a functional ATPbinding site as well as conserved interactions with the channel-forming subunit FutB in the transporter complex. Polyamines are important for the cell proliferation, differentiation and apoptosis in prokaryotic and eukaryotic cells. In plants, polyamines have special roles in stress response and in plant survival. The polyamine metabolism in cyanobacteria in response to environmental stress is of interest in research on stress tolerance of higher plants. In this thesis, the potd gene encoding an polyamine transporter subunit from Synechocystis sp. strain PCC 6803 was characterized for the first time. A homology model built for PotD protein indicated that it has capability of binding polyamines, with the preference for spermidine. Furthermore, in order to investigate the structural features of the substrate specificity, polyamines were docked into the binding site. Spermidine was positioned very similarly in Synechocystis PotD as in the template structure and had most favorable interactions of the docked polyamines. Based on the homology model, experimental work was conducted, which confirmed the binding preference. Flavodiiron proteins (Flv) are enzymes, which protect the cell against toxicity of oxygen and/or nitric oxide by reduction. In this thesis, we present a novel type of photoprotection mechanism in cyanobacteria by the heterodimer of Flv2/Flv4. The constructed homology model of Flv2/Flv4 suggests a functional heterodimer capable of rapid electron transfer. The unknown protein sll0218, encoded by the flv2-flv4 operon, is assumed to facilitate the interaction of the Flv2/Flv4 heterodimer and energy transfer between the phycobilisome and PSII. Flv2/Flv4 provides an alternative electron transfer pathway and functions as an electron sink in PSII electron transfer.

On-site calibration of a phase fraction meter by an inverse technique

The formal calibration procedure of a phase fraction meter is based on registering the outputs resulting from imposed phase fractions at known flow regimes. This can be straightforwardly done in laboratory conditions, but is rarely the case in industrial conditions, and particularly for on-site applications. Thus, there is a clear need for less restrictive calibration methods regarding to the prior knowledge of the complete set of inlet conditions. A new procedure is proposed in this work for the on-site construction of the calibration curve from total flown mass values of the homogeneous dispersed phase. The solution is obtained by minimizing a convenient error functional, assembled with data from redundant tests to handle the intrinsic ill-conditioned nature of the problem. Numerical simulations performed for increasing error levels demonstrate that acceptable calibration curves can be reconstructed, even from total mass measured within a precision of up to 2%. Consequently, the method can readily be applied, especially in on-site calibration problems in which classical procedures fail due to the impossibility of having a strict control of all the input/output parameters.

Methods of genetic diversity creation and functional display for directed evolution experiments

Protein engineering aims to improve the properties of enzymes and affinity reagents by genetic changes. Typical engineered properties are affinity, specificity, stability, expression, and solubility. Because proteins are complex biomolecules, the effects of specific genetic changes are seldom predictable. Consequently, a popular strategy in protein engineering is to create a library of genetic variants of the target molecule, and render the population in a selection process to sort the variants by the desired property. This technique, called directed evolution, is a central tool for trimming protein-based products used in a wide range of applications from laundry detergents to anti-cancer drugs. New methods are continuously needed to generate larger gene repertoires and compatible selection platforms to shorten the development timeline for new biochemicals. In the first study of this thesis, primer extension mutagenesis was revisited to establish higher quality gene variant libraries in Escherichia coli cells. In the second study, recombination was explored as a method to expand the number of screenable enzyme variants. A selection platform was developed to improve antigen binding fragment (Fab) display on filamentous phages in the third article and, in the fourth study, novel design concepts were tested by two differentially randomized recombinant antibody libraries. Finally, in the last study, the performance of the same antibody repertoire was compared in phage display selections as a genetic fusion to different phage capsid proteins and in different antibody formats, Fab vs. single chain variable fragment (ScFv), in order to find out the most suitable display platform for the library at hand. As a result of the studies, a novel gene library construction method, termed selective rolling circle amplification (sRCA), was developed. The method increases mutagenesis frequency close to 100% in the final library and the number of transformants over 100-fold compared to traditional primer extension mutagenesis. In the second study, Cre/loxP recombination was found to be an appropriate tool to resolve the DNA concatemer resulting from error-prone RCA (epRCA) mutagenesis into monomeric circular DNA units for higher efficiency transformation into E. coli. Library selections against antigens of various size in the fourth study demonstrated that diversity placed closer to the antigen binding site of antibodies supports generation of antibodies against haptens and peptides, whereas diversity at more peripheral locations is better suited for targeting proteins. The conclusion from a comparison of the display formats was that truncated capsid protein three (p3Δ) of filamentous phage was superior to the full-length p3 and protein nine (p9) in obtaining a high number of uniquely specific clones. Especially for digoxigenin, a difficult hapten target, the antibody repertoire as ScFv-p3Δ provided the clones with the highest affinity for binding. This thesis on the construction, design, and selection of gene variant libraries contributes to the practical know-how in directed evolution and contains useful information for scientists in the field to support their undertakings.