Osakassopimus osakeyhtiön osakkaiden keskinäisten suhteiden sääntelyssä

Osakkeenomistajien keskinaisia suhteita sekä osakkeenomistajien suhdetta yhtiöön ja kolmanteen sääntelevät muun muassa osakeyhtiölain (29.9.1978/734) saannokset ja yhtiöjärjestyksen määräykset. Osakeyhtiölaissa on vain rajoitetusti otettu huomioon se, että osakeyhtiöiden tarkoitukset vaihtelevat ja että ne ovat erikokoisia. Saannokset soveltuvat parhaiten suuren tai keskisuuren osakeyhtiön osakaspiirin tarpeisiin. Osakassopimuksella pyritään tavallisesti siihen, että asiat olisivat ennen niiden käsittelemistä yhtiön elimissä jo edeltä kasin sovittu. Paatosten tekemistä ei kuitenkaan voida osakkeenomistajien keskinaisella sopimuksella siirtää osakeyhtiölaissa sääntelemättömille elimille. Osakassopimuksilla pyritään varmistamaan osakkaiden oikeudet yhtiön elinkaaren aikana muuttuvissa tilanteissa. Yhdistävänä tekijänä niissä on valta ja sen käyttö. Osakassopimukset ovat jaettavissa vallan keskittämistä, hajauttamista ja tasapainon ylläpitämistä koskeviin sopimuksiin. Osakassopimuksella saatetaan pyrkiä säilyttämään osakkeenomistajien osakassopimuksen tekohetken aikaiset vaikutusmahdollisuudet muuttumattomina heidän keskinaisissa suhteissaan ja suhteissa ulkopuolisiin intressiryhmiin.

Streptavidin - A Versatile Binding Protein for Solid-Phase Immunoassays

Streptavidin, a tetrameric protein secreted by Streptomyces avidinii, binds tightly to a small growth factor biotin. One of the numerous applications of this high-affinity system comprises the streptavidin-coated surfaces of bioanalytical assays which serve as universal binders for straightforward immobilization of any biotinylated molecule. Proteins can be immobilized with a lower risk of denaturation using streptavidin-biotin technology in contrast to direct passive adsorption. The purpose of this study was to characterize the properties and effects of streptavidin-coated binding surfaces on the performance of solid-phase immunoassays and to investigate the contributions of surface modifications. Various characterization tools and methods established in the study enabled the convenient monitoring and binding capacity determination of streptavidin-coated surfaces. The schematic modeling of the monolayer surface and the quantification of adsorbed streptavidin disclosed the possibilities and the limits of passive adsorption. The defined yield of 250 ng/cm² represented approximately 65 % coverage compared with a modelled complete monolayer, which is consistent with theoretical surface models. Modifications such as polymerization and chemical activation of streptavidin resulted in a close to 10-fold increase in the biotin-binding densities of the surface compared with the regular streptavidin coating. In addition, the stability of the surface against leaching was improved by chemical modification. The increased binding densities and capacities enabled wider high-end dynamic ranges in the solid-phase immunoassays, especially when using the fragments of the capture antibodies instead of intact antibodies for the binding of the antigen. The binding capacity of the streptavidin surface was not, by definition, predictive of the low-end performance of the immunoassays nor the assay sensitivity. Other features such as non-specific binding, variation and leaching turned out to be more relevant. The immunoassays that use a direct surface readout measurement of time-resolved fluorescence from a washed surface are dependent on the density of the labeled antibodies in a defined area on the surface. The binding surface was condensed into a spot by coating streptavidin in liquid droplets into special microtiter wells holding a small circular indentation at the bottom. The condensed binding area enabled a denser packing of the labeled antibodies on the surface. This resulted in a 5 - 6-fold increase in the signal-to-background ratios and an equivalent improvement in the detection limits of the solid-phase immunoassays. This work proved that the properties of the streptavidin-coated surfaces can be modified and that the defined properties of the streptavidin-based immunocapture surfaces contribute to the performance of heterogeneous immunoassays.

Highly functional binding surface for miniaturised solid-phase immunoassays. - A spot story -

Several bioaffinity assays are based on the detection of an analyte which is bound on a solid substrate via biochemical interaction. These so called solid phase assays are based on the adhesion of the primary binding partner on a solid surface, which then binds the analyte to be detected. In this thesis work a novel solid phase based assay technology, known as spot technology, was developed. The spot technology is based on combination of high-capacity solid phases, concentrated in a spot format, utilising modified streptavidin molecules and recombinant antibody fragments. The reduction of the solid phase binding surface to a size of a spot enabled denser binding of the target molecules, providing improved signal intensities and signal-to-background ratio when applied in different solid phase immunoassays. Streptavidin-biotin interactions are commonly utilised in numerous different bioaffinity assays and the ultimate nature of streptavidin to bind biotin is among the strongest non-covalent interaction reported between two biomolecules. In this study native core streptavidin was chemically modified to provide polymerised streptavidin molecules with altered adsorption properties. These streptavidin conjugates, when coated onto polystyrene surface, provided enhanced biotin binding capacity and surface stability when compared to a reference coating constructed with native streptavidin. Furthermore, the combination of chemically modified streptavidin, sitespecifically biotinylated antibody fragments and the spot coating technology provided highly dense solid phase coating with improved binding properties. The performance of the spot assay technology was further demonstrated in different immunoassay configurations. Human thyroid stimulating hormone (TSH) and human cardiac troponin I (cTnI) were used as model analytes to show the applicability of the highly sensitive spot-based solid-phase immunoassay for detection of very low levels of analytes. It was demonstrated that the spot technology provided an assay concept with enhanced sensitivity and short turn-around times, characteristics that are highly suitable for point-of-care applications.

Contract manufacturing networks structure and supply chain management

The usual objectives that companies have for subcontracting are studied in this thesis. The case company’s objectives for contract manufacturing now and in the future are identified. The main objective of the thesis is to create a focused model for the structure and supply chain management in the contract manufacturing network. This model is made for case company’s certain profit center. The different possibilities and their advantages and disadvantages for the structure and supply chain management are examined trough a theoretical review of literature. The possibilities found are then examined from the case company’s point of view. The case company point of view is established based on the opinions of the case company’s representatives. The outcome of the thesis is that the star shaped structure with supply chain management centralized to case company would be the best choice for the case company to manage the contract manufacture network.

Comparing service contract prices of outsourcing partner

The main objective of this Master’s thesis is to find out which one of the two pricing models is the most cost-effective. In this thesis there are two companies that have made an outsourcing contract, in which they have a possibility to choose between two different pricing models. The first model is so called FTE (Full Time Employee) -based. The total cost will be based on the amount of outsourced person-workyears. The second pricing model is the transaction-based, in which the price will be formed according to the amount of transactions. Changing the pricing model from FTE-based to the transaction-based will also incur other costs. It is very important that these other costs are also taken into consideration, so that it is possible to determine the total costs of the pricing models. These other costs are direct costs, indirect costs and performance related costs of outsourcing. Activity based-costing (ABC) was used in order to find out the trues indirect costs of the outsourced processes. Performance related costs are related to quality, so Pareto-analysis was used to analyse the costs. Based on all of that, a framework for service related cost analysis was developed. Quality costs were almost impossible to quantify, so quality had to be taken into consideration in a qualitative way. Furthermore, considering only the indirect and direct costs in a quantitative way and quality costs in a qualitative way, it was possible to find a conditional solution for the research question.

Conformational Regulation of alpha2beta1 Integrin in Ligand Binding

Integrins are heterodimeric cell adhesion receptors involved in cell-cell and cell-extracellular matrix (ECM) interactions. They transmit bidirectional signals across the cell membrane. This results in a wide range of biological events from cell differentiation to apoptosis. alpha2beta1 integrin is an abundant collagen receptor expressed on the surface of several cell types. In addition to ECM ligands, alpha2beta1 integrins are bound by echovirus 1 (EV1) which uses alpha2beta1 as a receptor to initiate its life cycle in the infected cell. The aim of this thesis project was to provide further insight into the mechanisms of alpha2beta1 integrin ligand recognition and receptor activation. Collagen fibrils are the principal tensile elements of the ECM. Yet, the interaction of alpha2beta1 integrin with the fibrillar form of collagen I has received relatively little attention. This research focused on the ability of alpha2beta1 integrin to act as a receptor for type I collagen fibrils. Also the molecular requirements of the EV1 interaction with alpha2beta1 were studied. Conventionally, ligand binding has been suggested to require integrin activation and the binding may further trigger integrin signalling. Another main objective of this study was to elucidate both the inside-out and outside-in signalling mechanisms of alpha2beta1 integrin in adherent cells. The results indicated that alpha2beta1 integrin is the principal integrin-type collagen receptor for type I collagen fibrils, and alpha2beta1 may participate in the regulation of pericellular collagen fibrillogenesis. Furthermore, alpha2beta1 integrin inside-out activation appeared to be synergistically regulated by integrin clustering and conformational activation. The triggering of alpha2beta1 integrin outside-in signalling, however, was shown to require both conformational changes and clustering. In contrast to ECM ligands, EV1 appeared to take advantage of the bent, inactive form of alpha2beta1 integrin in initiating its life cycle in the cell. This research together with other recent studies, has shed light on the molecular mechanisms of integrin activation. It is becoming evident that large ligands are able to bind to the bent form of integrin, which has been previously considered to be physiologically inactive. Consequently, our understanding of the conformational modulation of integrins upon activation is changing.

Sopimuksen muuttaminen liiketoimintasuhteessa

Tutkielman tavoitteena on selvittää sopimuksen muuttamiseen liittyviä kysymyksiä liiketoimintasuhteessa. Lähtökohtana kysymyksenasettelulle on kansallinen lainsäädäntö, sopimusoikeudelliset periaatteet sekä oikeuskirjallisuus. Sopimuksen muuttamisen liikesuhteissa mahdollistaa Suomen lainsäädännössä OikTL 36 §, jonka johdosta kohtuutonta sopimusta on mahdollista sovitella. Sovittelun mahdollisuus heikentää kuitenkin sopimuksen sitovuutta, sillä lähtökohtaisesti sopimus sitoo sovituilla ehdoilla. Sopimuksen muutostarpeen yhteydessä sopimuksen tulkinta ja osapuolten tarkoitus nousevat avainasemaan tarkasteltaessa edellytyksiä kohtuuttomuuteen vetoamiseen ja sovitteluun. Lain ohella merkityksellisessä asemassa ovat siten sopimukseen otetut ehdot ja sopimusrakenteet.

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.

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.

Excelling in procurement : IT Contract Management with Lean and Six Sigma. Case study on legacy contract management at Philips Global IT.

Few people see both opportunities and threats coming from IT legacy in current world. On one hand, effective legacy management can bring substantial hard savings and smooth transition to the desired future state. On the other hand, its mismanagement contributes to serious operational business risks, as old systems are not as reliable as it is required by the business users. This thesis offers one perspective of dealing with IT legacy – through effective contract management, as a component towards achieving Procurement Excellence in IT, thus bridging IT delivery departments, IT procurement, business units, and suppliers. It developed a model for assessing the impact of improvements on contract management process and set of tools and advices with regards to analysis and improvement actions. The thesis conducted case study to present and justify the implementation of Lean Six Sigma in IT legacy contract management environment. Lean Six Sigma proved to be successful and this thesis presents and discusses all the steps necessary, and pitfalls to avoid, to achieve breakthrough improvement in IT contract management process performance. For the IT legacy contract management process two improvements require special attention and can be easily copied to any organization. First is the issue of diluted contract ownership that stops all the improvements, as people do not know who is responsible for performing those actions. Second is the contract management performance evaluation tool, which can be used for monitoring, identifying outlying contracts and opportunities for improvements in the process. The study resulted in a valuable insight on the benefits of applying Lean Six Sigma to improve IT legacy contract management, as well as on how Lean Six Sigma can be applied in IT environment. Managerial implications are discussed. It is concluded that the use of data-driven Lean Six Sigma methodology for improving the existing IT contract management processes is a significant addition to the existing best practices in contract management.

Service Productization and Product Servitization in The Contract Manufacturing Company

The role of contract manufacturing and subcontracting has been seen in black and white in product and service point of view. It used to be seen either as a product or a service. In the thesis product-service system, offering combining products and services, was discussed. Theory was created from two perspectives; Service productization via Business Model generation and product servitization via New Service Development process. Target for the case study was to point out new ways of service thinking and ways for changing business environment in contract manufacturing, especially in customer satisfaction and profitability point of view. The case study is following the New Service Development process phases. First ideas were collected from literature and via sales management interviews. Service offering and tool for service requirement evaluation was created. Last financial results of example service scenarios were calculated. It is recommended to take service offering into internal use and further develop it into modular service model. It is also recommended to take created customer service requirement evaluation tool into use for capturing customer service needs but also for communicating those internally.

Muscarinic toxins : characterization of adrenoceptor binding and applicability of membrane anchoring via glycosylphosphatidylinositol tail

Adrenoceptors (ARs), G-protein coupled receptors (GPCRs) at the plasma membrane, respond to endogenous catecholamines noradrenaline and adrenaline. These receptors mediate several important physiological functions being especially important in the cardiovascular system and in the regulation of smooth muscle contraction. Impairments in the function of these receptors can thus lead to severe diseases and disorders such as to cardiovascular diseases and benign prostatic hyperplasia. The Eastern green mamba (Dendroaspis angusticeps) venom has been shown to contain toxins that can antagonize the functions of GPCRs. The most well-known are muscarinic toxins (MTs) targeting muscarinic acetylcholine receptors (mAChRs) with high affinity and selectivity. However, some reports have indicated that these toxins might also act on the α1- and α2-ARs which can be divided into various subtypes; the α1-ARs to α1A-, α1B- and α1D-ARs and α2-ARs to α2A-, α2B- and α2C-ARs. In this thesis, the interaction of four common MTs (MT1, MT3, MT7 and MTα) with the adrenoceptors was characterized. It was also evaluated whether these toxins could be anchored to the plasma membrane via glycosylphosphatidylinositol (GPI) tail. Results of this thesis reveal that muscarinic toxins are targeting several α-adrenoceptor subtypes in addition to their previously identified target receptors, mAChRs. MTα was found to interact with high affinity and selectivity with the α2B-AR whereas MT7 confirmed its selectivity for the M1 mAChR. Unlike MTα and MT7, MT1 and MT3 have a broad range of target receptors among the α-ARs. All the MTs characterized were found to behave as non-competitive antagonists of receptor action. The interaction between MTα and the α2B-AR was studied more closely and it was observed that the second extracellular loop of the receptor functions as a structural entity enabling toxin binding. The binding of MTα to the α2B-AR appears to be rather complex and probably involves dimerized receptor. Anchoring MTs to the plasma membrane did not interfere with their pharmacological profile; all the GPI-anchored toxins created retained their ability to block their target receptors. This thesis shows that muscarinic toxins are able to target several subtypes of α-ARs and mAChRs. These toxins offer thus a possibility to create new subtype specific ligands for the α-AR subtypes. Membrane anchored MTs on the other hand could be used to block α-AR and mAChR actions in disease conditions such as in hypertension and in gastrointestinal and urinary bladder disorders in a cell-specific manner and to study the physiological functions of ARs and mAChRs in vivo in model organisms.