Measurement of Bioactivity of Immunomodulatory Treatment of Relapsing Multiple Sclerosis. With Emphasis on MxA Protein

Background Multiple sclerosis (MS) is a demyelinating disease of the central nervous system, which mainly affects young adults. In Finland, approximately 2500 out of 6000 MS patients have relapsing MS and are treated with disease modifying drugs (DMD): interferon- β (INF-β-1a or INF-β-1b) and glatiramer acetate (GA). Depending on the used IFN-β preparation, 2 % to 40 % of patients develop neutralizing antibodies (NAbs), which abolish the biological effects of IFN-β, leading to reduced clinical and MRI detected efficacy. According to the Finnish Current Care Guidelines and European Federation of Neurological Societis (EFNS) guidelines, it is suggested tomeasure the presence of NAbs during the first 24 months of IFN-β therapy. Aims The aim of this thesis was to measure the bioactivity of IFN-β therapy by focusing on the induction of MxA protein (myxovirus resistance protein A) and its correlation to neutralizing antibodies (NAb). A new MxA EIA assay was set up to offer an easier and rapid method for MxA protein detection in clinical practice. In addition, the tolerability and safety of GA were evaluated in patients who haddiscontinued IFN-β therapy due to side effects and lack of efficacy. Results NAbs developed towards the end of 12 months of treatment, and binding antibodies were detectable before or parallel with them. The titer of NAb correlated negatively with the amount of MxA protein and the mean values of preinjection MxA levels never returned to true baseline in NAb negative patients, but tended to drop in the NAb positive group. The test results between MxA EIA and flow cytometric analysis showed significant correlation. GA reduced the relapse rate and was a safe and well-tolerated therapy in IFN-β-intolerant MS patients. Conclusions NAbs inhibit the induction of MxA protein, which can be used as a surrogate marker of the bioactivity of IFN-β therapy. Compared to flow cytometricanalysis and NAb assay, MxA-EIA seemed to be a sensitive and more practical method in clinical use to measure the actual bioactivity of IFN-β treatment, which is of value also from a cost-effective perspective.

A comparative performance study of four-pole induction motors and synchronous reluctance motors in variable speed drives

Design aspects of the Transversally Laminated Anisotropic (TLA) Synchronous Reluctance Motor (SynRM) are studied and the machine performance analysis compared to the Induction Motor (IM) is done. The SynRM rotor structure is designed and manufactured for a30 kW, four-pole, three-phase squirrel cage induction motor stator. Both the IMand SynRM were supplied by a sensorless Direct Torque Controlled (DTC) variablespeed drive. Attention is also paid to the estimation of the power range where the SynRM may compete successfully with a same size induction motor. A technicalloss reduction comparison between the IM and SynRM in variable speed drives is done. The Finite Element Method (FEM) is used to analyse the number, location and width of flux barriers used in a multiple segment rotor. It is sought for a high saliency ratio and a high torque of the motor. It is given a comparison between different FEM calculations to analyse SynRM performance. The possibility to take into account the effect of iron losses with FEM is studied. Comparison between the calculated and measured values shows that the design methods are reliable. A new application of the IEEE 112 measurement method is developed and used especially for determination of stray load losses in laboratory measurements. The study shows that, with some special measures, the efficiency of the TLA SynRM is equivalent to that of a high efficiency IM. The power factor of the SynRM at rated load is smaller than that of the IM. However, at lower partial load this difference decreases and this, probably, brings that the SynRM gets a better power factor in comparison with the IM. The big rotor inductance ratio of the SynRM allows a good estimating of the rotor position. This appears to be very advantageous for the designing of the rotor position sensor-less motor drive. In using the FEM designed multi-layer transversally laminated rotor with damper windings it is possible to design a directly network driven motor without degrading the motorefficiency or power factor compared to the performance of the IM.

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.

Fouling management and retention in nanofiltration of integrated paper mill effluents

Nanofiltration performance was studied with effluents from the pulp and paper industry and with model substances. The effect of filtration conditions and membrane properties on nanofiltration flux, retention, and fouling was investigated. Generally, the aim was to determine the parameters that influence nanofiltration efficiency and study how to carry out nanofiltration without fouling by controlling these parameters. The retentions of the nanofiltration membranes studied were considerably higher than those of tight ultrafiltration membranes, and the permeate fluxes obtained were approximately the same as those of tight ultrafiltration membranes. Generally, about 80% retentions of total carbon and conductivity were obtained during the nanofiltration experiments. Depending on the membrane and the filtration conditions, the retentions of monovalent ions (chloride) were between 80 and 95% in the nanofiltrations. An increase in pH improved retentions considerably and also the flux to some degree. An increase in pressure improved retention, whereas an increase in temperature decreased retention if the membrane retained the solute by the solution diffusion mechanism. In this study, more open membranes fouled more than tighter membranes due to higher concentration polarization and plugging of the membrane material. More irreversible fouling was measured for hydrophobic membranes. Electrostatic repulsion between the membrane and the components in the solution reduced fouling but did not completely prevent it with the hydrophobic membranes. Nanofiltration could be carried out without fouling, at least with the laboratory scale apparatus used here when the flux was below the critical flux. Model substances had a strong form of the critical flux, but the effluents had only a weak form of the critical flux. With the effluents, some fouling always occurred immediately when the filtration was started. However, if the flux was below the critical flux, further fouling was not observed. The flow velocity and pH were probably the most important parameters, along with the membrane properties, that influenced the critical flux. Precleaning of the membranes had only a small effect on the critical flux and retentions, but it improved the permeability of the membranes significantly.

Prognostic Markers and Prolonged Interferon-alpha Therapy in Renal Cell Carcinoma

Molekyylimarkkerit ja pitkäaikainen alfainterferonihoito munuaissyövässä Munuaissyöpäpotilaiden viiden vuoden elossaololuku on noin 50 %. Aikaisempien tutkimuksien mukaan viiden vuoden elossaololuku metastasoituneessa munuaissyövässä on 3-16 %, kun käytettiin alfainterferonia sisältävää hoitoa. Tyypillisesti alfainterferonia on käytetty vähemmäin kuin 6 kuukautta. Avoimia kysymyksiä ovat alfainterferonin optimaalinen hoitoannos ja hoidon kesto yksin tai yhdessä uusien täsmähoitojen kanssa. Tärkeimmät tavoitteet olivat tutkia 1) jaksotetun pitkäaikaisen alfainterferonihoidon tehoa ja siedettävyyttä metastasoituneessa munuaissyövässä ja 2) p53-, Ki-67- ja COX-2-proteiinituotannon ennusteellista merkitystä munuaissyövässä. Tutkimuksessa 117 metastasoituneelle munuaissyöpää sairastaneelle potilaalle etsittiin yksilöllinen hänen sietämänsä maksimaalinen hoitoannos rekombinanttia alfa2a-interferonia (Roferon-ATM). Hoitoa pyrittiin jatkamaan 24 kuukauden ajan. Kolmen hoitoviikon jälkeen pidettiin yhden viikon tauko. Hoito lopetettiin, jos ilmaantui vakavia haittavaikutuksia tai tauti eteni. Toisessa tutkimuksessa proteiinituotanto analysoitiin immunohistokemiallisesti munuaissyöpäpotilaiden kasvainnäytteistä, joita oli säilytetty parafiinissa. Kasvainnäytteet oli otettu talteen munuaisen poistoleikkauksen yhteydessä. Nämä potilaat jaettiin kolmeen eri ryhmään: metastasointi primaarivaiheessa (n=29), metastasointi myöhemmin (n=37) ja ei metastasointia (n=51). Keskimääräinen alfainterferonihoidon kesto oli 11 kuukautta (kk) [0,5 – 32 kk]. Objektiivinen hoitovaste todettiin 17 %:lla, tautitilanne pysyi ennallaan 42 %:lla ja myöhäinen vaste (yli 12 kk:tta hoidon aloittamisesta) todettiin 3 %:lla. Aika vasteen saavuttamisesta taudin etenemiseen oli keskimäärin 8 kk ja elinaika 19,1 kk. Viiden vuoden elossaololuku oli 16 %. Jos metastasoituneella munuaissyöpäpotilaalla oli keuhkometastasointi, hän selvisi todennäköisemmin viisi vuotta kuin muut potilaat. Henkeä uhkaavia sivuvaikutuksia ei todettu. Yli 12 kk:n ajan kestävä alfainterferonihoito on hyödyllistä niille potilaille, jotka ovat saaneet objektiivisen hoitovasteen tai tautitilanne on pysynyt ennallaan. Positiivinen p53- ja Ki-67-ekspressio yhdessä viittaavat suureen metastasoinnin todennäköisyyteen. Positiivinen COX-2-ekspressio viittaa viivästyneeseen metastaasien ilmaantumiseen. Metastasoituneilla potilailla positiiviset p53- ja Ki-67-ekspressiot viittaavat huonoon ennusteeseen, mutta positiivinen COX-2 ekspressio viittaa suotuisaan ennusteeseen. Positiivinen COX-2- ja negatiivinen Ki-67-ekspressio yhdessä viittaavat parantuneeseen ennusteeseen metastasoituneessa munuaissyövässä.

Molecular profiling of human endometrium and endometriosis

Endometriosis is a common hormone-dependent gynecological disease leading to severe menstrual and/or chronic pelvic pain with or without subfertility. The disease is defined by the presence of endometrium-like tissue outside the uterine cavity, primarily on the pelvic peritoneum, ovaries and infiltrating organs of the peritoneal cavity. The current tools for diagnosis and treatment of endometriosis need to be improved to ensure reliable diagnosis and effective treatment. In addition, endometriosis is associated with increased risk of ovarian cancer and, therefore, the differential diagnosis between the benign and malignant ovarian cysts is of importance. The long-term objective of the present study was to support the discovery of novel tools for diagnosis and treatment of endometriosis. This was approached by exploiting genome-wide expression analysis of endometriosis specimens. A novel expression profiling -based classification of endometriosis indicated specific subgroups of lesions partially consistent with the clinical appearance, but partially according to unknown factors. The peritoneum of women with endometriosis appeared to be altered in comparison to that of healthy control subjects, suggesting a novel aspect on the pathogenesis of the disease. The evaluation of action and metabolism of sex hormones in endometrium and endometriosis tissue indicated a novel role of androgens in regulation of the tissues. In addition, an enzyme involved in androgen and neurosteroid metabolism, hydroxysteroid (17beta) dehydrogenase 6, was found to be highly up-regulated in endometriosis tissue as compared to healthy endometrium. The enzyme may have a role in the pathogenesis of endometriosis or in the endometriosis associated pain generation. Finally, a new diagnostic biomarker, HE4, was discovered distinguishing patients with ovarian endometriotic cysts from those with malignant ovarian cancer. The information acquired in this study enables deeper understanding of endometriosis and facilitates the development of improved diagnostic tools and more specific treatments of the disease

Online Ultrasound Measurements of Membrane Compaction

There are several filtration applications in the pulp and paper industry where the capacity and cost-effectiveness of processes are of importance. Ultrafiltration is used to clean process water. Ultrafiltration is a membrane process that separates a certain component or compound from a liquid stream. The pressure difference across the membrane sieves macromolecules smaller than 0.001-0.02 μm through the membrane. When optimizing the filtration process capacity, online information about the conditions of the membrane is needed. Fouling and compaction of the membrane both affect the capacity of the filtration process. In fouling a “cake” layer starts to build on the surface of the membrane. This layer blocks the molecules from sieving through the membrane thereby decreasing the yield of the process. In compaction of the membrane the structure is flattened out because of the high pressure applied. The higher pressure increases the capacity but may damage the structure of the membrane permanently. Information about the compaction is needed to effectively operate the filters. The objective of this study was to develop an accurate system for online monitoring of the condition of the membrane using ultrasound reflectometry. Measurements of ultrafiltration membrane compaction were made successfully utilizing ultrasound. The results were confirmed by permeate flux decline, measurements of compaction with a micrometer, mechanical compaction using a hydraulic piston and a scanning electron microscope (SEM). The scientific contribution of this thesis is to introduce a secondary ultrasound transducer to determine the speed of sound in the fluid used. The speed of sound is highly dependent on the temperature and pressure used in the filters. When the exact speed of sound is obtained by the reference transducer, the effect of temperature and pressure is eliminated. This speed is then used to calculate the distances with a higher accuracy. As the accuracy or the resolution of the ultrasound measurement is increased, the method can be applied to a higher amount of applications especially for processes where fouling layers are thinner because of smaller macromolecules. With the help of the transducer, membrane compaction of 13 μm was measured in the pressure of 5 bars. The results were verified with the permeate flux decline, which indicated that compaction had taken place. The measurements of compaction with a micrometer showed compaction of 23–26 μm. The results are in the same range and confirm the compaction. Mechanical compaction measurements were made using a hydraulic piston, and the result was the same 13 μm as obtained by applying the ultrasound time domain reflectometry (UTDR). A scanning electron microscope (SEM) was used to study the structure of the samples before and after the compaction.

Influenza in children – Diagnosis, treatment and prevention

Background: The burden of influenza on children is substantial. Although mortality rates are low, the incidence of influenza is highest in children, among whom also complications are frequent. A more accurate recognition of influenza in children could enable the rational use of antiviral drugs and help to avoid unnecessary courses of antibiotics. Limited data exists on the efficacy of oseltamivir treatment and the trivalent inactivated influenza vaccine (TIV) in children. Aims and methods: We sought for signs and symptoms that could help clinicians to diagnose influenza on clinical grounds in a case-control study in children <13 years of age. We further assessed the feasibility of different diagnostics methods during the early stage of the illness in children aged 1-3 years. The efficacy of early oseltamivir treatment (started <24h from the onset of symptoms) was evaluated in a randomized controlled trial (RCT) conducted in children 1-3 years of age, and the effectiveness of TIV to prevent laboratory-confirmed influenza was determined in a prospective, observational cohort study conducted among children aged 9 months to 3 years of age. Results: Fever was the only symptom predicting influenza in children. The sensitivity of conventionally used laboratory methods to detect influenza during the first 24h of illness was 92%. The sensitivity of the influenza rapid test in the same setting was 90% for influenza A and 25% for influenza B. Early oseltamivir treatment shortened the duration of the illness in children with influenza A by 3.5-4.0 days, but no efficacy was observed against influenza B. The effectiveness of TIV was 84% against the wellmatched influenza A, while no effectiveness against the mismatched influenza B was observed. Conclusions: Laboratory diagnostics are needed for a reliable diagnosis of influenza in children and were found sensitive already during the early stage of the illness. Early oseltamivir treatment was highly effective against influenza A, but no efficacy was seen against influenza B. TIV is effective also in young children if a good match between the vaccine and circulating strain is achieved.

Treatment of Acute Otitis Media

Background: Acute otitis media (AOM) is the most common bacterial infection in young children, but the optimal management of AOM remains controversial. The aim of this study was to assess the efficacy of antimicrobial treatment, either immediate or delayed, for AOM and to compare parental experiences regarding the management of AOM in two countries with very different treatment guidelines. Methods: Altogether, 322 children participated in a randomized, double-blind, placebocontrolled trial. Children 6–35 months of age with AOM received amoxicillin-clavulanate or placebo for 7 days. The primary outcome was the time to treatment failure. In the second study, the delayed antimicrobial treatment group consisted of recipients of placebo who had received rescue treatment. The immediate antimicrobial treatment group consisted of children allocated to amoxicillin-clavulanate group. Parental expectations and opinions were evaluated by questionnaires sent via public day care in Turku, Finland, and Utrecht, the Netherlands. Results: Treatment failure occurred significantly more often in children receiving placebo as compared to antimicrobial treatment (45% vs. 19%, P<0.001). Delayed initiation of antimicrobial treatment did not worsen the recovery from AOM, but it was associated with worsening of the child’s condition, prolongation of symptoms, and absenteeism from day care and parental absenteeism from work. According to the comparative questionnaire, antimicrobial use was more common in Finland than in the Netherlands. Finnish parents believed more often than Dutch parents that antimicrobials are necessary in the treatment of AOM. Conclusions: Children with AOM benefit from antimicrobial treatment. Delayed initiation of antimicrobial does not worsen the overall recovery from AOM, but it might increase the symptom burden and create economic losses. Treatment practices and parental expectations seem to interact with each other. This needs to be considered when AOM treatment guidelines are updated.

Situational awareness concept in a multinational collaboration environment : challenges in the information sharing framework

Technological capabilities are built to support different types of collaboration, and this gives the justification to widely observe, how activity environments are influenced by technology. Technology as an enabler can be addressed from different perspectives, other than merely technological. Dynamic, evolving environment is at the same time interesting but also challenging. As a multinational collaboration environment, the maritime surveillance is an good example of time critical and evolving environment, where technological solutions enable new ways of collaboration. Justification for the inspiration to use maritime environment as the baseline for understanding the challenges in creating and maintaining adequate level of situational awareness, derives from the complexity of the collaboration and information sharing environment elements, needed to be taken into account, when analyzing criticalities related to decision making. Situational awareness is an important element supporting decision making, and challenges related to it can also be observed in the maritime environment. This dissertation describes the structures and factors involved in this complex setting, found from the case studies that should be taken into account when trying to understand, how these elements affect the activities. This dissertation focuses on the gray area that is between a life threatening situation and normal everyday activities. From the multinational experimentation series case studies, MNE5 and MNE6 it was possible to observe situations that were not life threatening for the participants themselves, but not also basic every day activities. These case studies provided a unique possibility to see situations, where gaining of situational awareness and decision making are challenged with time critical crisis situations. Unfortunately organizations do not normally take the benefit from the everyday work to prepare themselves for possible emerging crisis situations. This dissertation focuses on creating a conceptual model and a concept that supports organizations – also outside the maritime community – to improve their ability to support gaining of situational awareness from the individual training level, all the way to changes in organizational structures in aiming for better support for decision making from the individual level to the highest decision making level. Quick changes and unpredictability are reality in organizations and organizations do not have the possibility to control all the factors that affect their functioning. Since we cannot be prepared for everything, and predict every crisis, individual activities inside teams and as a part of organizations, need to be supported with guidance, tools and training in order to support acting in challenging situations. In fact the ideology of the conceptual model created, lies especially in the aim of not controlling everything in beforehand, but supporting organizations with concrete procedures to help individuals to react in different, unpredictable situations, instead of focusing on traditional risk prevention and management. Technological capabilities are not automatically solutions for functional challenges; this is why it is justified to broaden the problem area observation from the technological perspective. This dissertation demonstrates that it is possible to support collaboration in a multinational environment with technological solutions, but it requires the recognition of technological limitations and accepting the possible restrictions related to technological innovations. Technology should not be considered value per se, the value of technology should be defined according to the support of activities, including strategic and operational environment evaluation, identification of organizational elements, and taking into account also the social factors and their challenges. Then we are one step closer to providing technological solutions that support the actual activities by taking into account the variables of the activity environment in question. The multidisciplinary view to approach the information sharing and collaboration framework, is derived especially from the complexity of decision making and building of situational awareness, since they are not build or created in vacuity, but in the organizational framework by the people doing it with the technological capabilities, enabled by the organizational structures. Introduced case studies were related to maritime environment, but according to the research results, it is valid to argue, that based on the lessons learned it is possible to create and further develop conceptual model and to create a general concept to support a wider range of organizations in their attempt to gain better level of situational awareness (SA) and to support decision making. To proof the versatile usage of the developed concept, I have introduced the case study findings to the health care environment and reflected the identified elements from the trauma center to the created concept. The main contribution to complete this adventure is the presented situational awareness concept created in the respect to NATO concept structure. This has been done to tackle the challenge of collaboration by focusing on situational awareness in the information sharing context by providing a theoretical ground and understanding, of how these issues should be approached, and how these elements can be generalized and used to support activities in other environments as well. This dissertation research has been a several year evolving process reflecting and affecting presented case studies and this learning experience from the case studies has also affected the goals and research questions of this dissertation. This venture has been written from a retro perspective according to ideology of process modeling and design rationale to present to the reader how this entire journey took place and what where the critical milestones that affected the end result, conceptual model. Support in a challenging information sharing framework can be provided with the right type of combination of tools, procedures and individual effort. This dissertation will provide insights to those with a new approach to war technology for the organizations to gain a better level of awareness and to improve the capabilities in decision making. This dissertation will present, from the war technology starting point, a new approach and possibility for the organizations to create a better level of awareness and support for decision making with the right combination of tools, procedures and individual effort.

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.

Liquid cooling solutions for rotating permanent magnet synchronous machines

In the design of electrical machines, efficiency improvements have become very important. However, there are at least two significant cases in which the compactness of electrical machines is critical and the tolerance of extremely high losses is valued: vehicle traction, where very high torque density is desired at least temporarily; and direct-drive wind turbine generators, whose mass should be acceptably low. As ever higher torque density and ever more compact electrical machines are developed for these purposes, thermal issues, i.e. avoidance of over-temperatures and damage in conditions of high heat losses, are becoming of utmost importance. The excessive temperatures of critical machine components, such as insulation and permanent magnets, easily cause failures of the whole electrical equipment. In electrical machines with excitation systems based on permanent magnets, special attention must be paid to the rotor temperature because of the temperature-sensitive properties of permanent magnets. The allowable temperature of NdFeB magnets is usually significantly less than 150 ˚C. The practical problem is that the part of the machine where the permanent magnets are located should stay cooler than the copper windings, which can easily tolerate temperatures of 155 ˚C or 180 ˚C. Therefore, new cooling solutions should be developed in order to cool permanent magnet electrical machines with high torque density and because of it with high concentrated losses in stators. In this doctoral dissertation, direct and indirect liquid cooling techniques for permanent magnet synchronous electrical machines (PMSM) with high torque density are presented and discussed. The aim of this research is to analyse thermal behaviours of the machines using the most applicable and accurate thermal analysis methods and to propose new, practical machine designs based on these analyses. The Computational Fluid Dynamics (CFD) thermal simulations of the heat transfer inside the machines and lumped parameter thermal network (LPTN) simulations both presented herein are used for the analyses. Detailed descriptions of the simulated thermal models are also presented. Most of the theoretical considerations and simulations have been verified via experimental measurements on a copper tooth-coil (motorette) and on various prototypes of electrical machines. The indirect liquid cooling systems of a 100 kW axial flux (AF) PMSM and a 110 kW radial flux (RF) PMSM are analysed here by means of simplified 3D CFD conjugate thermal models of the parts of both machines. In terms of results, a significant temperature drop of 40 ̊C in the stator winding and 28 ̊C in the rotor of the AF PMSM was achieved with the addition of highly thermally conductive materials into the machine: copper bars inserted in the teeth, and potting material around the end windings. In the RF PMSM, the potting material resulted in a temperature decrease of 6 ̊C in the stator winding, and in a decrease of 10 ̊C in the rotor embedded-permanentmagnets. Two types of unique direct liquid cooling systems for low power machines are analysed herein to demonstrate the effectiveness of the cooling systems in conditions of highly concentrated heat losses. LPTN analysis and CFD thermal analysis (the latter being particularly useful for unique design) were applied to simulate the temperature distribution within the machine models. Oil-immersion cooling provided good cooling capability for a 26.6 kW PMSM of a hybrid vehicle. A direct liquid cooling system for the copper winding with inner stainless steel tubes was designed for an 8 MW directdrive PM synchronous generator. The design principles of this cooling solution are described in detail in this thesis. The thermal analyses demonstrate that the stator winding and the rotor magnet temperatures are kept significantly below their critical temperatures with demineralized water flow. A comparison study of the coolant agents indicates that propylene glycol is more effective than ethylene glycol in arctic conditions.

Cholesterol, cardiovascular disease and statin use in older persons

Older age increases the risk of developing a chronic atherosclerotic cardiovascular disease (CVD), such as coronary heart disease. Complications of CVDs, myocardial infarction or stroke often lead to loss of functional capacity or premature death. Dyslipidemia, high serum levels of total or low-density lipoprotein cholesterol (LDL-c) and low levels of high-density lipoprotein cholesterol (HDL-c), is among the most important modifiable risk factors for CVDs; it can be treated with lifestyle modifications, and with lipid-lowering drugs, primarily statins. In older persons, however, the association of cholesterol levels with cardiovascular and all-cause mortality has been inconsistent in previous studies. Furthermore, the beneficial effects of statins in older persons without previous CVD are still somewhat unclear, and older persons are more prone to adverse effects from statins. This thesis presents a prospective cohort study (TUVA), exploring associations of cholesterol levels with mortality and the changes in cholesterol levels of a 70-year-old population in long-term follow-up. Further, prevalence of CVDs, risk factors and preventive medication use in the TUVA cohort is compared with respective prevalences in another age-matched cohort (UTUVA) 20 years later in order to examine the changes in cardiovascular risk over time. Additionally, to evaluate statin use patterns among older persons, an observational register study was conducted covering the total Finnish population aged 70 and older during 2000-2008. Based on individual-level data retrieved from national health registries, the population was classified into low, moderate and high risk groups according to estimated CVD risk. The prevalence, incidence and persistence of statin use among the risk groups was then evaluated based upon yearly statin purchases tracked from the Prescription Register. The prospective cohort study demonstrated that low total cholesterol, LDL-c and HDL-c were associated with higher mortality in a cohort of home-dwelling 70-year-olds. However, after adjusting for traditional cardiovascular risk factors and cancer this association disappeared. Further, low total cholesterol seemed to be protective, whereas low HDL-c strongly predicted increased risk of CVD death. Cholesterol levels of those elderly who remained available for follow-up and were still home-dwelling at the age of 85 seemed to improve with advancing age. Compared to the TUVA cohort, the later born UTUVA cohort had less CVDs and their risk factors were better controlled, which was reflected in the higher use of preventive medications such as statins and antihypertensives. The register studies confirmed that statin use has increased significantly during 2000-2008 among older persons, especially among the oldest (80+) age groups and among those at high risk for cardiovascular events. Two-thirds of new statin users persisted with their use during the four years of follow-up; the most discontinuations were made during the first year of use. In conclusion, statins are commonly used among older age groups in Finland. Most of the older statin users had a high cardiovascular event risk, indicating that the treatment is well directed towards those who are likely to benefit from it the most. No age-limits should be put on the screening and treatment of dyslipidemia in older persons, but the benefits and adverse effects of statin treatment should be carefully weighed based on an individual assessment of the person’s general health status and functional capacity. Physicians should pay more attention to medication adherence, especially when prescribing preventive medications.