Alpha2-adrenoceptors: Structure and ligand binding properties at the molecular level

Alpha₂-Adrenoceptors: structure and ligand binding properties at the molecular level The mouse is the most frequently used animal model in biomedical research, but the use of zebrafish as a model organism to mimic human diseases is on the increase. Therefore it is considered important to understand their pharmacological differences from humans also at the molecular level. The zebrafish Alpha_2-adrenoceptors were expressed in mammalian cells and the binding affinities of 20 diverse ligands were determined and compared to the corresponding human receptors. The pharmacological properties of the human and zebrafish Alpha_2--adrenoceptors were found to be quite well conserved. Receptor models based on the crystal structures of bovine rhodopsin and the human Beta₂-adrenoceptor revealed that most structural differences between the paralogous and orthologous Alpha_2--adrenoceptors were located within the second extracellular loop (XL2). Reciprocal mutations were generated in the mouse and human Alpha_2--adrenoceptors. Ligand binding experiments revealed that substitutions in XL2 reversed the binding profiles of the human and mouse Alpha_2--adrenoceptors for yohimbine, rauwolscine and RS-79948-197, evidence for a role for XL2 in the determination of species-specific ligand binding. Previous mutagenesis studies had not been able to explain the subtype preference of several large Alpha_2--adrenoceptor antagonists. We prepared chimaeric Alpha_2--adrenoceptors where the first transmembrane (TM1) domain was exchanged between the three human Alpha_2--adrenoceptor subtypes. The binding affinities of spiperone, spiroxatrine and chlorpromazine were observed to be significantly improved by TM1 substitutions of the Alpha_2a--adrenoceptor. Docking simulations indicated that indirect effects, such as allosteric modulation, are more likely to be involved in this phenomenon rather than specific side-chain interactions between ligands and receptors.

Landscape structure in the northern coast of Paraná state, a hotspot for the brazilian Atlantic Forest conservation

The "Serra do Mar" region comprises the largest remnant of the Brazilian Atlantic Forest. The coast of the Paraná State is part of the core area of the "Serra do Mar" corridor and where actions for biodiversity conservation must be planned. In this study we aimed at characterizing the landscape structure in the APA-Guaraqueçaba, the largest protected area in this region, in order to assist environmental policies of this region. Based on a supervised classification of a mosaic of LANDSAT-5-TM satellite images (from March 2009), we developed a map (1:75,000 scale) with seven classes of land use and land cover and analyzed the relative quantities of forests and modified areas in slopes and lowlands. The APA-Guaraqueçaba is comprised mainly by the Dense Ombrophilous Forest (68.6% of total area) and secondary forests (9.1%), indicating a forested landscape matrix; anthropogenic and bare soil areas (0.8%) and the Pasture/Grasslands class (4.2%) were less representative. Slopes were less fragmented and more preserved (96.3% of Dense Ombrophilous Forest and secondary forest) than lowlands (71.3%), suggesting that restoration initiatives in the lowlands must be stimulated in this region. We concluded that most of the region sustains well-conserved ecosystems, highlighting the importance of Paraná northern coast for the biodiversity maintenance of the Atlantic Forest.

Making Sense of Shared Leadership. A case study of leadership processes and practices without formal leadership structure in the team context

Leadership is essential for the effectiveness of the teams and organizations they are part of. The challenges facing organizations today require an exhaustive review of the strategic role of leadership. In this context, it is necessary to explore new types of leadership capable of providing an effective response to new needs. The presentday situations, characterized by complexity and ambiguity, make it difficult for an external leader to perform all leadership functions successfully. Likewise, knowledge-based work requires providing professional groups with sufficient autonomy to perform leadership functions. This study focuses on shared leadership in the team context. Shared leadership is seen as an emergent team property resulting from the distribution of leadership influence across multiple team members. Shared leadership entails sharing power and influence broadly among the team members rather than centralizing it in the hands of a single individual who acts in the clear role of a leader. By identifying the team itself as a key source of influence, this study points to the relational nature of leadership as a social construct where leadership is seen as social process of relating processes that are co-constructed by several team members. Based on recent theoretical developments concerned with relational, practice-based and constructionist approaches to the study of leadership processes, this thesis proposes the study of leadership interactions, working processes and practices to focus on the construction of direction, alignment and commitment. During the research process, critical events, activities, working processes and practices of a case team have been examined and analyzed with the grounded theory –approach in the terms of shared leadership. There are a variety of components to this complex process and a multitude of factors that may influence the development of shared leadership. The study suggests that the development process of shared leadership is a common sense -making process and consists of four overlapping dimensions (individual, social, structural, and developmental) to work with as a team. For shared leadership to emerge, the members of the team must offer leadership services, and the team as a whole must be willing to rely on leadership by multiple team members. For these individual and collective behaviors to occur, the team members must believe that offering influence to and accepting it from fellow team members are welcome and constructive actions. Leadership emerges when people with differing world views use dialogue and collaborative learning to create spaces where a shared common purpose can be achieved while a diversity of perspectives is preserved and valued. This study also suggests that this process can be supported by different kinds of meaning-making and process tools. Leadership, then, does not reside in a person or in a role, but in the social system. The built framework integrates the different dimensions of shared leadership and describes their relationships. This way, the findings of this study can be seen as a contribution to the understanding of what constitutes essential aspects of shared leadership in the team context that can be of theoretical value in terms of advancing the adoption and development process of shared leadership. In the real world, teams and organizations can create conditions to foster and facilitate the process. We should encourage leaders and team members to approach leadership as a collective effort that the team can be prepared for, so that the response is rapid and efficient.

Genetic structure and diversity of Copaifera langsdorffii Desf. in Cerrado fragments of the São Paulo State, Brazil

The loss of large areas of Cerrado (Brazilian savanna) in Brazil can lead to reduced biodiversity and to the extinction of species. Therefore, the present study aimed to investigate the genetic fragility of populations of Copaifera langsdorffii Desf exposed to different anthropic conditions in fragments of Cerrado in the state of São Paulo. The study was carried out in two Experimental Stations operated by the Forest Institute (Assis and Itirapina), in one fully protected conservation unit (Pedregulho) and in one private property (Brotas). Analyses were conducted using leaf samples from 353 adult specimens and eight pairs of microsatellite loci. The number of alleles per locus ranged from 13 to 15 in all populations, but the mean number of effective alleles was approximately half this value (7.2 to 9-1). Observed heterozygosity was significant and lower than the expected in all populations. Consequently, all populations deviated from Hardy-Weinberg expected frequencies. Fixation indexes were significant for all populations, with the Pedregulho population having the lowest value (0.189) and Itirapina having the highest (0.283). The analysis of spatial genetic structure detected family structures at distance classes of 20 to 65 m in the populations studied. No clones were detected in the populations. Estimates of effective population size were low, but the area occupied by each population studied was large enough for conservation, medium and long term. Recent reductions or bottlenecks were detected in all four populations. Mean Gst’ (genetic divergence) indicated that most of the variation was within populations. Cluster structure analysis based on the genotypes detected K= 4 clusters with distinct allele frequencies patterns. The genetic differentiation observed among populations is consistent with the hypothesis of genetic and geographic isolation. Therefore, it is essential to adopt conservation strategies that raise the gene flow between fragments.

Frequency-Domain and Wide-Pulse Time-Domain Measurements of Lanthanide Luminescence and Lanthanide-Based Resonance Energy Transfer

Resonance energy transfer (RET) is a non-radiative transfer of the excitation energy from the initially excited luminescent donor to an acceptor. The requirements for the resonance energy transfer are: i) the spectral overlap between the donor emission spectrum and the acceptor absorption spectrum, ii) the close proximity of the donor and the acceptor, and iii) the suitable relative orientations of the donor emission and the acceptor absorption transition dipoles. As a result of the RET process the donor luminescence intensity and the donor lifetime are decreased. If the acceptor is luminescent, a sensitized acceptor emission appears. The rate of RET depends strongly on the donor–acceptor distance (r) and is inversely proportional to r6. The distance dependence of RET is utilized in binding assays. The proximity requirement and the selective detection of the RET-modified emission signal allow homogeneous separation free assays. The term lanthanide-based RET is used when luminescent lanthanide compounds are used as donors. The long luminescence lifetimes, the large Stokes’ shifts and the intense, sharply-spiked emission spectra of the lanthanide donors offer advantages over the conventional organic donor molecules. Both the organic lanthanide chelates and the inorganic up-converting phosphor (UCP) particles have been used as donor labels in the RET based binding assays. In the present work lanthanide luminescence and lanthanide-based resonance energy transfer phenomena were studied. Luminescence lifetime measurements had an essential role in the research. Modular frequency-domain and time-domain luminometers were assembled and used successfully in the lifetime measurements. The frequency-domain luminometer operated in the low frequency domain ( 100 kHz) and utilized a novel dual-phase lock-in detection of the luminescence. One of the studied phenomena was the recently discovered non-overlapping fluorescence resonance energy transfer (nFRET). The studied properties were the distance and temperature dependences of nFRET. The distance dependence was found to deviate from the Förster theory and a clear temperature dependence was observed whereas conventional RET was completely independent of the temperature. Based on the experimental results two thermally activated mechanisms were proposed for the nFRET process. The work with the UCP particles involved the measurement of the luminescence properties of the UCP particles synthesized in our laboratory. The goal of the UCP particle research is to develop UCP donor labels for binding assays. In the present work the effect of the dopant concentrations and the core–shell structure on the total up-conversion luminescence intensity, the red–green emission ratio, and the luminescence lifetime was studied. Also the non-radiative nature of the energy transfer from the UCP particle donors to organic acceptors was demonstrated for the first time in aqueous environment and with a controlled donor–acceptor distance.

Influence of stiffness in bolted connections in wooden plane structure of truss type

Trusses are structural systems commonly used in projects, being employed mainly in roof structures, present in most rural buildings. The design of trusses, as well as other structural systems, requires the determination of displacements, strains and stresses. However, the project is developed from an ideal model of calculation, considering free rotation between the elements of a connection. This paper presents a computer program for the analysis of bidimensional wooden trusses with connections formed with two screws per node. The formulation is based on the flexibility method, taking into account the influence of the effect of semi-rigid connections formed by two screws. An example of a structure is presented and analyzed by the program developed here, highlighting the importance of behavior analysis on semi-rigid connections.

Probabilistic structure of an annual extreme rainfall series of a coastal area of the State of São Paulo, Brazil

This study aimed to describe the probabilistic structure of the annual series of extreme daily rainfall (Preabs), available from the weather station of Ubatuba, State of São Paulo, Brazil (1935-2009), by using the general distribution of extreme value (GEV). The autocorrelation function, the Mann-Kendall test, and the wavelet analysis were used in order to evaluate the presence of serial correlations, trends, and periodical components. Considering the results obtained using these three statistical methods, it was possible to assume the hypothesis that this temporal series is free from persistence, trends, and periodicals components. Based on quantitative and qualitative adhesion tests, it was found that the GEV may be used in order to quantify the probabilities of the Preabs data. The best results of GEV were obtained when the parameters of this function were estimated using the method of maximum likelihood. The method of L-moments has also shown satisfactory results.

Improving Transportation and Warehousing Efficiency with Simulation-Based Decision Support Systems

Transportation and warehousing are large and growing sectors in the society, and their efficiency is of high importance. Transportation also has a large share of global carbondioxide emissions, which are one the leading causes of anthropogenic climate warming. Various countries have agreed to decrease their carbon emissions according to the Kyoto protocol. Transportation is the only sector where emissions have steadily increased since the 1990s, which highlights the importance of transportation efficiency. The efficiency of transportation and warehousing can be improved with the help of simulations, but models alone are not sufficient. This research concentrates on the use of simulations in decision support systems. Three main simulation approaches are used in logistics: discrete-event simulation, systems dynamics, and agent-based modeling. However, individual simulation approaches have weaknesses of their own. Hybridization (combining two or more approaches) can improve the quality of the models, as it allows using a different method to overcome the weakness of one method. It is important to choose the correct approach (or a combination of approaches) when modeling transportation and warehousing issues. If an inappropriate method is chosen (this can occur if the modeler is proficient in only one approach or the model specification is not conducted thoroughly), the simulation model will have an inaccurate structure, which in turn will lead to misleading results. This issue can further escalate, as the decision-maker may assume that the presented simulation model gives the most useful results available, even though the whole model can be based on a poorly chosen structure. In this research it is argued that simulation- based decision support systems need to take various issues into account to make a functioning decision support system. The actual simulation model can be constructed using any (or multiple) approach, it can be combined with different optimization modules, and there needs to be a proper interface between the model and the user. These issues are presented in a framework, which simulation modelers can use when creating decision support systems. In order for decision-makers to fully benefit from the simulations, the user interface needs to clearly separate the model and the user, but at the same time, the user needs to be able to run the appropriate runs in order to analyze the problems correctly. This study recommends that simulation modelers should start to transfer their tacit knowledge to explicit knowledge. This would greatly benefit the whole simulation community and improve the quality of simulation-based decision support systems as well. More studies should also be conducted by using hybrid models and integrating simulations with Graphical Information Systems.

Influence of Link Structure on Search Engine Result Pages of Online Nursery Shops

The primary objective of this thesis is to assess how the backlink portfolio structure and off site Search Engine Optimisation (SEO) elements influence ranking of UK based online nursery shops. The growth of the internet use demanded significant effort from companies to optimize and increase their online presence in order to cope with the increasing online competition. The new e-Commerce technology - called Search Engine Optimisation - has been developed that helped increase website visibility of companies. The SEO process involves on site elements (i.e. changing the parameters of the company's website such as keywords, title tags and meta descriptions) and off site elements (link building and social media marketing activity). Link Building is based on several steps of marketing planning including keyword research and competitor analysis. The underlying goal of keyword research is to understand the targeted market through identifying relevant keyword queries that are used by targeted costumer group. In the analysis, three types (geographic, field and company’s strategy related) and seven sources of keywords has been identified and used as a base of analysis. Following the determination of the most popular keywords, allinanchor and allintitle search has been conducted and the first ten results of the searches have been collected to identify the companies with the most significant web presence among the nursery shops. Finally, Link Profiling has been performed where the essential goal was to understand to what extent other companies' link structure is different that the base company's backlinks. Significant difference has been found that distinguished the top three companies ranking in the allinanchor and allintitle search. The top three companies, „Mothercare”, „Mamas and Papas” and „Kiddicare” maintained significantly better metrics regarding domain and page authority on the main landing pages, the average number of outbound links for link portfolio metric and in number of backlinks. These companies also ranked among the highest in page authority distribution and followed external linking.

Fatigue study of the fillet weld waving on weld toe and the penetration in the weld root based on the effective notch stress approach

The fatigue failure of structures under fluctuating loads in fillet weld joints raises a demand to determine the parameters related to this type of loading. In this study, the stress distribution in the susceptible area of weld toe and weld root in fillet welded models analyzed by finite element method applying FEMAP software. To avoid the geometrical singularity on the path of analytical stress analysis in the toe and root area of a weld model the effective notch stress approach applied by which a proper fictitious rounding that mostly depend on the material of structure is applied. The models with different weld toe waving width and radius are analyzed while the flank angle of weld varied in 45 and 30 degrees. The processed results shows that the waving compare to the straight weld toe makes differences in the value of stress and consequently the stress concentration factor between the tip and depth of the waves in the weld toe which helps to protect the crack of propagation and gives enough time and tools to be informed of the crack initiation in the structure during the periodical observation of structure. In the weld root study the analyses among the models with the welding penetration percentage from non-penetration to the full-penetration shows a slightly increase in the root area stress value which comparing with the stiffening effect of penetration conclude that the half-penetration can make an optimization between the stress increase and stiffening effect of deep penetration.

Quasiclassical Approach to the Vortex State in Iron-based Superconductors

The quasiclassical approach was applied to the investigation of the vortex properties in the ironbased superconductors. The special attention was paid to manifestation of the nonlocal effects of the vortex core structure. The main results are as follows: (i) The effects of the pairing symmetries (s+ and s₊₊) on the cutoff parameter of field distribution, ξh, in stoichiometric (like LiFeAs) and nonstoichiometric (like doped BaFe₂As₂) iron pnictides have been investigated using Eilenberger quasiclassical equations. Magnetic field, temperature and impurity scattering dependences of ξh have been calculated. Two opposite behavior have been discovered. The ξh /ξc2 ratio is less in s+ symmetry when intraband impurity scattering (Γ₀) is much larger than one and much larger than interband impurity scattering (Γπ), i.e. in nonstoichiometric iron pnictides. Opposite, the value ξh /ξc2 is higher in s+ case and the field dependent curve is shifted upward from the "clean" case (Γ₀ = Γπ = 0) for stoichiometric iron pnictides (Γ₀ = Γπ ≪ 1). (ii) Eilenberger approach to the cutoff parameter, ξh, of the field distribution in the mixed state of high

Power cycling lifetime estimation of IGBT power modules based on chip temperature modeling

In this doctoral thesis, methods to estimate the expected power cycling life of power semiconductor modules based on chip temperature modeling are developed. Frequency converters operate under dynamic loads in most electric drives. The varying loads cause thermal expansion and contraction, which stresses the internal boundaries between the material layers in the power module. Eventually, the stress wears out the semiconductor modules. The wear-out cannot be detected by traditional temperature or current measurements inside the frequency converter. Therefore, it is important to develop a method to predict the end of the converter lifetime. The thesis concentrates on power-cycling-related failures of insulated gate bipolar transistors. Two types of power modules are discussed: a direct bonded copper (DBC) sandwich structure with and without a baseplate. Most common failure mechanisms are reviewed, and methods to improve the power cycling lifetime of the power modules are presented. Power cycling curves are determined for a module with a lead-free solder by accelerated power cycling tests. A lifetime model is selected and the parameters are updated based on the power cycling test results. According to the measurements, the factor of improvement in the power cycling lifetime of modern IGBT power modules is greater than 10 during the last decade. Also, it is noticed that a 10 C increase in the chip temperature cycle amplitude decreases the lifetime by 40%. A thermal model for the chip temperature estimation is developed. The model is based on power loss estimation of the chip from the output current of the frequency converter. The model is verified with a purpose-built test equipment, which allows simultaneous measurement and simulation of the chip temperature with an arbitrary load waveform. The measurement system is shown to be convenient for studying the thermal behavior of the chip. It is found that the thermal model has a 5 C accuracy in the temperature estimation. The temperature cycles that the power semiconductor chip has experienced are counted by the rainflow algorithm. The counted cycles are compared with the experimentally verified power cycling curves to estimate the life consumption based on the mission profile of the drive. The methods are validated by the lifetime estimation of a power module in a direct-driven wind turbine. The estimated lifetime of the IGBT power module in a direct-driven wind turbine is 15 000 years, if the turbine is located in south-eastern Finland.

The Structure and Function of αI Domains in Collagen Receptor and Leukocyte Integrins

Integrins are heterodimeric, signaling transmembrane adhesion receptors that connect the intracellular actin microfilaments to the extracellular matrix composed of collagens and other matrix molecules. Bidirectional signaling is mediated via drastic conformational changes in integrins. These changes also occur in the integrin αI domains, which are responsible for ligand binding by collagen receptor and leukocyte specific integrins. Like intact integrins, soluble αI domains exist in the closed, low affinity form and in the open, high affinity form, and so it is possible to use isolated αI domains to study the factors and mechanisms involved in integrin activation/deactivation. Integrins are found in all mammalian tissues and cells, where they play crucial roles in growth, migration, defense mechanisms and apoptosis. Integrins are involved in many human diseases, such as inflammatory, cardiovascular and metastatic diseases, and so plenty of effort has been invested into developing integrin specific drugs. Humans have 24 different integrins, four of which are collagen receptor (α1β1, α2β1, α10β1, α11β1) and five leukocyte specific integrins (αLβ2, αMβ2, αXβ2, αDβ2, αEβ7). These two integrin groups are quite unselective having both primary and secondary ligands. This work presents the first systematic studies performed on these integrin groups to find out how integrin activation affects ligand binding and selectivity. These kinds of studies are important not only for understanding the partially overlapping functions of integrins, but also for drug development. In general, our results indicated that selectivity in ligand recognition is greatly reduced upon integrin activation. Interestingly, in some cases the ligand binding properties of integrins have been shown to be cell type specific. The reason for this is not known, but our observations suggest that cell types with a higher integrin activation state have lower ligand selectivity, and vice versa. Furthermore, we solved the three-dimensional structure for the activated form of the collagen receptor α1I domain. This structure revealed a novel intermediate conformation not previously seen with any other integrin αI domain. This is the first 3D structure for an activated collagen receptor αI domain without ligand. Based on the differences between the open and closed conformation of the αI domain we set structural criteria for a search for effective collagen receptor drugs. By docking a large number of molecules into the closed conformation of the α2I domain we discovered two polyketides, which best fulfilled the set structural criteria, and by cell adhesion studies we showed them to be specific inhibitors of the collagen receptor integrins.

The impact of a new partnership outsourcing arrangement: a routine-based case study in detail engineering

This thesis investigated the contemporary phenomenon of detail engineering outsourcing. The case organization had pursued a new outsourcing approach with a trusted partner. The goal of this empirical study was to examine the impact of the consequential partnership outsourcing arrangement. Particularly, the beneficence of the arrangement was evaluated based on the underlying organizational routine and the long-term economic implications of its performance outcome. The case study was needed, as the unit will likely have to rely on such distance outsourcing arrangements more and more in the future, and understanding on the impact of such operations is needed. The main findings revealed that the new outsourcing arrangement is not currently a very attractive strategic option for organizing production. The benefits which stem from the emerged, unique engineering project routine are not significant enough to make the arrangement an advantageous one, especially since increasing partnering costs are being met. This conclusion was drawn via the extended transaction cost view. Benchmarking was done in reliance to an old arrangement from which the new pursuit was a departure from. The case study then enlightened the engineering unit on the impact of its strategic maneuver by combining the routines-theory framework with contemporary methods of governance structure evaluation. Through this, it was shown that greater efforts are needed to make the new outsourcing approach a more beneficial one. However, the studied arrangement was seen to inhold potential for better results. The findings can be used to capitalize on this.