Phylogenetic Analysis of Avidins and Expression of Novel Avidins from Lactrodectus hesperus and Hoeflea phototrophica

Avidins (Avds) are homotetrameric or homodimeric glycoproteins with typically less than 130 amino acid residues per monomer. They form a highly stable, non-covalent complex with biotin (vitamin H) with Kd = 10-15 M (for chicken Avd). The best-studied Avds are the chicken Avd from Gallus gallus and streptavidin from Streptomyces avidinii, although other Avd studies have also included Avds from various origins, e.g., from frogs, fishes, mushrooms and from many different bacteria. Several engineered Avds have been reported as well, e.g., dual-chain Avds (dcAvds) and single-chain Avds (scAvds), circular permutants with up to four simultaneously modifiable ligand-binding sites. These engineered Avds along with the many native Avds have potential to be used in various nanobiotechnological applications. In this study, we made a structure-based alignment representing all currently available sequences of Avds and studied the evolutionary relationship of Avds using phylogenetic analysis. First, we created an initial multiple sequence alignment of Avds using 42 closely related sequences, guided by the known Avd crystal structures. Next, we searched for non-redundant Avd sequences from various online databases, including National Centre for Biotechnology Information and the Universal Protein Resource; the identified sequences were added to the initial alignment to expand it to a final alignment of 242 Avd sequences. The MEGA software package was used to create distance matrices and a phylogenetic tree. Bootstrap reproducibility of the tree was poor at multiple nodes and may reflect on several possible issues with the data: the sequence length compared is relatively short and, whereas some positions are highly conserved and functional, others can vary without impinging on the structure or the function, so there are few informative sites; it may be that periods of rapid duplication have led to paralogs and that the differences among them are within the error limit of the data; and there may be other yet unknown reasons. Principle component analysis applied to alternative distance data did segregate the major groups, and success is likely due to the multivariate consideration of all the information. Furthermore, based on our extensive alignment and phylogenetic analysis, we expressed two novel Avds, lacavidin from Lactrodectus Hesperus, a western black widow spider, and hoefavidin from Hoeflea phototrophica, an aerobic marine bacterium, the ultimate aim being to determine their X-ray structures. These Avds were selected because of their unique sequences: lacavidin has an N-terminal Avd-like domain but a long C-terminal overhang, whereas hoefavidin was thought to be a dimeric Avd. Both these Avds could be used as novel scaffolds in biotechnological applications.

Analysis of the Change Management Process in a Multinational Company

The purpose of this thesis was to study change management and find out how the theory applies to practise with the help of studying an organizational change process. The goal was to investigate the different ways of the change management and how those could be utilized in practise. The purpose was also to see what kind of tools and information packages for change management the company involved has developed and taken in to use and check if these could have been utilized in the change process studied in this thesis. This thesis was established by studying the theory of change management and interviewing the people involved in the organizational change and the stakeholders. The questions were formed in line with the change management theory. The main theory used was John P. Kotter’s Eight Steps of Change. The study revealed valuable details about change processes in real life and concrete improvement ideas were recognized from the interviews. Overall the people were quite happy with the outcome of the change process. There were also some failures identified in this change process. Most probably those can be avoided in the future if people planning the change are familiar with the company’s new information packages and tools for change management. Change management is a complex area that is still today quite often forgotten by companies. Effective change management can give a huge competitive advantage for a company. Acknowledging that change process is always complex and not easy is already a good step forward in handling change processes. All changes need change management and understanding the way people react on change. A good and efficient change management is the key to make the change process smooth and easier for the people involved.

Offshoring and outsourcing under demand uncertainty -a supply chain management analysis

Outsourcing and offshoring or any combinations of these have not just become a popular phenomenon, but are viewed as one of the most important management strategies due to the new possibilities from globalization. They have been seen as a possibility to save costs and improve customer service. Executing offshoring and offshore outsourcing successfully can be more complex than initially expected. Potential cost savings resulting from of offshoring and offshore outsourcing are often based on lower manufacturing costs. However, these benefits might be conflicted by a more complex supply chain with service level challenges that can respectively increase costs. Therefore analyzing the total cost effects of offshoring and outsourcing is necessary. The aim of this Master´s Thesis was to to construct a total cost model using academic literature to calculate the total costs and analyze the reasonability of offshoring and offshore outsourcing production of a case company compared to insourcing production. The research data was mainly quantitative and collected mainly from the case company past sales and production records. In addition management level interviews from the case company were conducted. The information from these interviews was used for the qualification of the necessary quantitative data and adding supportive information that could not be gathered from the quantitative data. Both data collection and analysis were guided by a theoretical frame of reference that was based on academic literature concerning offshoring and outsourcing, statistical calculation of demand and total costs. The results confirm the theories that offshoring and offshore outsourcing would reduce total costs as both offshoring and offshore outsourcing options result in lower total annual costs than insourcing mainly due to lower manufacturing costs. However, increased demand uncertainty would make the alternative of offshore outsourcing more risky and difficult to manage. Therefore when assessing the overall impact of the alternatives, offshoring is the most preferable option. As the main cost savings in offshore outsourcing came from lower manufacturing costs, more specifically labour costs, the logistics costs in this case company did not have an essential effect in total costs. The management should therefore pay attention initially to manufacturing costs and then logistics costs when choosing the best production sourcing option for the company.

Software for Transport Accessibility Analysis: The Case of Moscow

In this thesis the process of building a software for transport accessibility analysis is described. The goal was to create a software which is easy to distribute and simple to use for the user without particular background in the field of the geographical data analysis. It was shown that existing tools do not suit for this particular task due to complex interface or significant rendering time. The goal was accomplished by applying modern approaches in the process of building web applications such as maps based on vector tiles, FLUX architecture design pattern and module bundling. It was discovered that vector tiles have considerable advantages over image-based tiles such as faster rendering and real-time styling.

FT-IR and MAS NMR analysis of montmorillonite K10 supported MF2 reagents and their activity as catalysis /

ZnF2, CdF2, and CUF2 have been adsorbed onto the surface of montmorillonite K10, and the infrared and 19F, 27 AI, and 29Si MAS NMR spectra of the reagents over a range of loadings have been obtained. CUF2 was observed to attack the Si02 layer and form the complex CuSiF6, Zn F2 tends to attack the aluminium oxide layer, in which Zn isomorphously replaces AI, and forms AIF3 and AIF4 - complexes. All the spectroscopic evidence ruled out the formation of any AI-F and/or Si-F free species as CdF2 is adsorbed on the surface of montmorillonite K10. The reactivity of MF2-K10 reagents towards Friedel-Crafts benzylation of benzene with benzyl chloride varied from one reagent to another. ZnF2-K10 was observed to be the most reactive and CUF2 was the least reactive.

Reactivity characteristics of cytochrome c, cytochrome oxidase and the electrostatic cytochrome c - cytochrome oxidase complex /

The mechanism whereby cytochrome £ oxidase catalyses elec-. tron transfer from cytochrome £ to oxygen remains an unsolved problem. Polarographic and spectrophotometric activity measurements of purified, particulate and soluble forms of beef heart mitochondrial cytochrome c oxidase presented in this thesis confirm the following characteristics of the steady-state kinetics with respect to cytochrome £: (1) oxidation of ferrocytochrome c is first order under all conditions. -(2) The relationship between sustrate concentration and velocity is of the Michaelis-Menten type over a limited range of substrate. concentrations at high ionic strength. (3) ~he reaction rate is independent from oxygen concentration until very low levels of oxygen. (4) "Biphasic" kinetic plots of enzyme activity as a function of substrate concentration are found when the range of cytochrome c concentrations is extended; the biphasicity ~ is more apparent in low ionic strength buffer. These results imply two binding sites for cytochrome £ on the oxidase; one of high affinity and one of low affinity with Km values of 1.0 pM and 3.0 pM, respectively, under low ionic strength conditions. (5) Inhibition of the enzymic rate by azide is non-c~mpetitive with respect to cytochrome £ under all conditions indicating an internal electron transfer step, and not binding or dissociation of £ from the enzyme is rate limiting. The "tight" binding of cytochrome '£ to cytochrome c oxidase is confirmed in column chromatographic experiments. The complex has a cytochrome £:oxidase ratio of 1.0 and is dissociated in media of high ionic strength. Stopped-flow spectrophotometric studies of the reduction of equimolar mixtures and complexes of cytochrome c and the oxidase were initiated in an attempt to assess the functional relevance of such a complex. Two alternative routes -for reduction of the oxidase, under conditions where the predominant species is the £ - aa3 complex, are postulated; (i) electron transfer via tightly bound cytochrome £, (ii) electron transfer via a small population of free cytochrome c interacting at the "loose" binding site implied from kinetic studies. It is impossible to conclude, based on the results obtained, which path is responsible for the reduction of cytochrome a. The rate of reduction by various reductants of free cytochrome £ in high and low ionic strength and of cytochrome £ electrostatically bound to cytochrome oxidase was investigated. Ascorbate, a negatively charged reagent, reduces free cytochrome £ with a rate constant dependent on ionic strength, whereas neutral reagents TMPD and DAD were relatively unaffected by ionic strength in their reduction of cytochrome c. The zwitterion cysteine behaved similarly to uncharged reductants DAD and TI~PD in exhibiting only a marginal response to ionic strength. Ascorbate reduces bound cytochrome £ only slowly, but DAD and TMPD reduce bound cytochrome £ rapidly. Reduction of cytochrome £ by DAD and TMPD in the £ - aa3 complex was enhanced lO-fold over DAD reduction of free £ and 4-fold over TMPD reduction of free c. Thus, the importance of ionic strength on the reactivity of cytochrome £ was observed with the general conclusion being that on the cytochrome £ molecule areas for anion (ie. phosphate) binding, ascorbate reduction and complexation to the oxidase overlap. The increased reducibility for bound cytochrome £ by reductants DAD and TMPD supports a suggested conformational change of electrostatically bound c compare.d to free .£. In addition, analysis of electron distribution between cytochromes £ and a in the complex suggest that the midpotential of cytochrome ~ changes with the redox state of the oxidase. Such evidence supports models of the oxidase which suggest interactions within the enzyme (or c - enzyme complex) result in altered midpoint potentials of the redox centers.

Structural analysis of the Paint Lake Deformation Zone, Northern Ontario /

The Paint Lake Deformation Zone (PLDZ), located within the Superior Province of Canada, demarcates a major structural and lithological break between the Onaman-Tashota Terrane to the north and the Beardmore-Geraldton Belt to the south. The PLDZ is an east-west trending lineament, approximately 50 km in length and up to 1 km in width, comprised of an early ductile component termed the Paint Lake Shear Zone and a late brittle component known as the Paint Lake Fault. Structures associated with PLDZ development including S-, C- and C'-fabrics, stretching lineations, slickensides, C-C' intersection lineations, Z-folds and kinkbands indicate that simple shear deformation dominated during a NW-SE compressional event. Movement along the PLDZ was in a dextral sense consisting of an early differential motion with southside- down and a later strike-slip motion. Although the locus of the PLDZ may in part be lithologically controlled, mylonitization which accompanied shear zone development is not dependent on the lithological type. Conglomerate, intermediate and mafic volcanic units exhibit similar mesoscopic and microscopic structures where transected by the PLDZ. Field mapping, supported by thin section analysis, defines five strain domains increasing in intensity of deformation from shear zone boundary to centre. A change in the dominant microstructural deformation mechanism from dislocation creep to diffusion creep is observed with increasing strain during mylonitization. C'-fabric development is temporally associated with this change. A decrease in the angular relationship between C- and C'-fabrics is observed upon attaining maximum strain intensity. Strain profiling of the PLDZ demonstrates the presence of an outer primary strain gradient which exhibits a simple profile and an inner secondary strain gradient which exhibits a more complex profile. Regionally metamorphosed lithologies of lower greenschist facies outside the PLDZ were subjected to retrograde metamorphism during deformation within the PLDZ.

Concept analysis in social studies

This project was undertaken because of a need to analyse concepts in social science more specifically and sequence them more carefully in a social science program. Concepts have been identified vaguely on many curriculum documents or left in isolation from each other when they are specifically identified. The project's aim was to identify a method for analysing concepts and sequencing their teaching on some rational basis. Once the method for analysing concepts was identified a questionnaire was designed and administered to a random sample of students at the grade three, five and eight levels. The questionnaire attempted to measure their comprehension of specific social science concepts at several levels which became progressively more complex. The major hypothesis was that there would be a direct correlation between age and achievement on the questionnaire. The raw scores were seriated and correlated with the ages of the students using the rankdifference- squared method. For the majority of areas tested it was found that there was a significant correlation between age and achievement on the questionnaire. Variation in the correlation coefficients generated suggests that comprehension of social science concepts is not simply a function of age but is probably a function of several inter-related factors such as reading ability, skill in Basic Thinking Skills and age. Thirty students completed each test. There were three tests in the questionnaire.

Rotation-electronic interaction in the 3p-complex Rydberg state of water

An energy theory is formulated for the rotational energy levels in a p-complex Rydberg state of an asymmetric top molecule of symmetry C2v. The effective Hamiltonian used consists of the usual rigid rotor Hamiltonian augmented with terms representing electronic spin and orbital angular momentum effects. Criteria for assigning symmetry species to the rotational energy levels, following Houganfs scheme that uses the full molecular group,are established and given in the form of a table. This is particularly suitable when eigenvectors are calculated on a digital computer. Also, an intensity theory for transitions to the Rydberg p-complex singlet states is presented and selection rules in terms of symmetry species of energy states are established. Finally, applications to HpO and DpO are given.

The Ontario Municipal Freedom of Information and Protection of Individual Privacy Act: an analysis of its application to school boards

In Canada freedom of information must be viewed in the context of governing -- how do you deal with an abundance of information while balancing a diversity of competing interests? How can you ensure people are informed enough to participate in crucial decision-making, yet willing enough to let some administrative matters be dealt with in camera without their involvement in every detail. In an age when taxpayers' coalition groups are on the rise, and the government is encouraging the establishment of Parent Council groups for schools, the issues and challenges presented by access to information and protection of privacy legislation are real ones. The province of Ontario's decision to extend freedom of information legislation to local governments does not ensure, or equate to, full public disclosure of all facts or necessarily guarantee complete public comprehension of an issue. The mere fact that local governments, like school boards, decide to collect, assemble or record some information and not to collect other information implies that a prior decision was made by "someone" on what was important to record or keep. That in itself means that not all the facts are going to be disclosed, regardless of the presence of legislation. The resulting lack of information can lead to public mistrust and lack of confidence in those who govern. This is completely contrary to the spirit of the legislation which was to provide interested members of the community with facts so that values like political accountability and trust could be ensured and meaningful criticism and input obtained on matters affecting the whole community. This thesis first reviews the historical reasons for adopting freedom of information legislation, reasons which are rooted in our parliamentary system of government. However, the same reasoning for enacting such legislation cannot be applied carte blanche to the municipal level of government in Ontario, or - ii - more specifially to the programs, policies or operations of a school board. The purpose of this thesis is to examine whether the Municipal Freedom of Information and Protection of Privacy Act, 1989 (MFIPPA) was a neccessary step to ensure greater openness from school boards. Based on a review of the Orders made by the Office of the Information and Privacy Commissioner/Ontario, it also assesses how successfully freedom of information legislation has been implemented at the municipal level of government. The Orders provide an opportunity to review what problems school boards have encountered, and what guidance the Commissioner has offered. Reference is made to a value framework as an administrative tool in critically analyzing the suitability of MFIPPA to school boards. The conclusion is drawn that MFIPPA appears to have inhibited rather than facilitated openness in local government. This may be attributed to several factors inclusive of the general uncertainty, confusion and discretion in interpreting various provisions and exemptions in the Act. Some of the uncertainty is due to the fact that an insufficient number of school board staff are familiar with the Act. The complexity of the Act and its legalistic procedures have over-formalized the processes of exchanging information. In addition there appears to be a concern among municipal officials that granting any access to information may be violating personal privacy rights of others. These concerns translate into indecision and extreme caution in responding to inquiries. The result is delay in responding to information requests and lack of uniformity in the responses given. However, the mandatory review of the legislation does afford an opportunity to address some of these problems and to make this complex Act more suitable for application to school boards. In order for the Act to function more efficiently and effectively legislative changes must be made to MFIPPA. It is important that the recommendations for improving the Act be adopted before the government extends this legislation to any other public entities.

Verbal ability, previous practice and load on short-term memory as determiners of differences in a complex learning task: an experimental study

Traditional psychometric theory and practice classify people according to broad ability dimensions but do not examine how these mental processes occur. Hunt and Lansman (1975) proposed a 'distributed memory' model of cognitive processes with emphasis on how to describe individual differences based on the assumption that each individual possesses the same components. It is in the quality of these components ~hat individual differences arise. Carroll (1974) expands Hunt's model to include a production system (after Newell and Simon, 1973) and a response system. He developed a framework of factor analytic (FA) factors for : the purpose of describing how individual differences may arise from them. This scheme is to be used in the analysis of psychometric tes ts . Recent advances in the field of information processing are examined and include. 1) Hunt's development of differences between subjects designated as high or low verbal , 2) Miller's pursuit of the magic number seven, plus or minus two, 3) Ferguson's examination of transfer and abilities and, 4) Brown's discoveries concerning strategy teaching and retardates . In order to examine possible sources of individual differences arising from cognitive tasks, traditional psychometric tests were searched for a suitable perceptual task which could be varied slightly and administered to gauge learning effects produced by controlling independent variables. It also had to be suitable for analysis using Carroll's f ramework . The Coding Task (a symbol substitution test) found i n the Performance Scale of the WISe was chosen. Two experiments were devised to test the following hypotheses. 1) High verbals should be able to complete significantly more items on the Symbol Substitution Task than low verbals (Hunt, Lansman, 1975). 2) Having previous practice on a task, where strategies involved in the task may be identified, increases the amount of output on a similar task (Carroll, 1974). J) There should be a sUbstantial decrease in the amount of output as the load on STM is increased (Miller, 1956) . 4) Repeated measures should produce an increase in output over trials and where individual differences in previously acquired abilities are involved, these should differentiate individuals over trials (Ferguson, 1956). S) Teaching slow learners a rehearsal strategy would improve their learning such that their learning would resemble that of normals on the ,:same task. (Brown, 1974). In the first experiment 60 subjects were d.ivided·into high and low verbal, further divided randomly into a practice group and nonpractice group. Five subjects in each group were assigned randomly to work on a five, seven and nine digit code throughout the experiment. The practice group was given three trials of two minutes each on the practice code (designed to eliminate transfer effects due to symbol similarity) and then three trials of two minutes each on the actual SST task . The nonpractice group was given three trials of two minutes each on the same actual SST task . Results were analyzed using a four-way analysis of variance . In the second experiment 18 slow learners were divided randomly into two groups. one group receiving a planned strategy practioe, the other receiving random practice. Both groups worked on the actual code to be used later in the actual task. Within each group subjects were randomly assigned to work on a five, seven or nine digit code throughout. Both practice and actual tests consisted on three trials of two minutes each. Results were analyzed using a three-way analysis of variance . It was found in t he first experiment that 1) high or low verbal ability by itself did not produce significantly different results. However, when in interaction with the other independent variables, a difference in performance was noted . 2) The previous practice variable was significant over all segments of the experiment. Those who received previo.us practice were able to score significantly higher than those without it. J) Increasing the size of the load on STM severely restricts performance. 4) The effect of repeated trials proved to be beneficial. Generally, gains were made on each successive trial within each group. S) In the second experiment, slow learners who were allowed to practice randomly performed better on the actual task than subjeots who were taught the code by means of a planned strategy. Upon analysis using the Carroll scheme, individual differences were noted in the ability to develop strategies of storing, searching and retrieving items from STM, and in adopting necessary rehearsals for retention in STM. While these strategies may benef it some it was found that for others they may be harmful . Temporal aspects and perceptual speed were also found to be sources of variance within individuals . Generally it was found that the largest single factor i nfluencing learning on this task was the repeated measures . What e~ables gains to be made, varies with individuals . There are environmental factors, specific abilities, strategy development, previous learning, amount of load on STM , perceptual and temporal parameters which influence learning and these have serious implications for educational programs .

A Foucaultian analysis of power relations in child protection case work with youth and families

This thesis takes some steps in examining the child protection system from a position that is rarely discussed. Specifically, I explore how Foucault's concept of disciplinary power can be used to demonstrate how power operates within the client/worker relationship. This relationship is shown to be quite complex with power flowing bidirectionally, rather than hierarchically. Instead of viewing power imbalances as a function of state control, I show how the client/worker relationship is constituted by the worker, the client, the organization and the social body. A postmodern auto ethnography is used to document my journey as I expose the disciplinary practices and instruments that I was subject to and used with my clients. 2 Given that the child protection system is constantly shifting and changing in order to improve its ability to safeguard children a greater emphasis is required to examine how workers operate within this complex, overwhelming and multi-dimensional world. This thesis has shown that by engaging in a reflexive examination of my position of power different approaches to making intervention beneficial to all involved become available. This is important if child protection work aims to work with clients rather than on clients.

Smart girls : a deconstructive discourse analysis of smartness

This project is a deconstructive discourse analysis of smart girlhood. From a feminist post structural framework, with a focus on discourse and performative identity, I scrutinize three dominant discourses of smartness that are prevalent and academic and popular press. These constructions frame smart girls as being either Losers, Have-It-All Girls, or Imposters. By conducting semi-structured group interviews with six self-identified smart girls, I explore the question of how smart girls perform their smart girl identities in their current sociocultural context. After analyzing the data from the group interviews, I outline five themes that seem to be prevalent in the stories told by the smart girls in this thesis. Finally, I discuss how the performative identities of the smart girls in my thesis appear to be much more complex, multiple and rhizomatic than the discourses under review allow.

Analysis of the Ethical, Social and Legal Issues Related to the Delivery of the Prenatal Genetic Testing in Argentina

L'introduction de nouvelles biotechnologies dans tout système de soins de santé est un processus complexe qui est étroitement lié aux facteurs économiques, politiques et culturels, et, par conséquent, demande de remettre en cause plusieurs questions sociales et éthiques. Dans la situation particulière de l’Argentine - c’est-à-dire: de grandes inégalités sociales entre les citoyens, la rareté des ressources sanitaires, l’accès limité aux services de base, l’absence de politiques spécifiques - l'introduction de technologies génétiques pose de sérieux défis qui doivent impérativement être abordés par les décideurs politiques. Ce projet examine le cas des tests génétiques prénataux dans le contexte du système de santé argentin pour illustrer comment leur introduction peut être complexe dans une nation où l’accès égale aux services de santé doit encore être amélioré. Il faut également examiner les restrictions légales et les préceptes religieux qui influencent l'utilisation des technologies génétiques, ce qui souligne la nécessite de développer un cadre de référence intégral pour le processus d'évaluation des technologies afin d’appuyer l’élaboration de recommandations pour des politiques cohérentes et novatrices applicables au contexte particulier de l’Argentine.

José Antonio Gómez’s Ynvitatorio, Himno y 8 Responsorios : historical context and music analysis of a manuscript ; José Antonio Gómez y Olguín, Ynvitatorio, Himno y 8 Responsorios, orchestral score

La version intégrale de cette thèse est disponible uniquement pour consultation individuelle à la Bibliothèque de musique de l’Université de Montréal (www.bib.umontreal.ca/MU).