Teollisuusyrityksen ekstranet –ratkaisut asiakaspalvelutoiminnan kehittämiseksi

Työssä tarkastellaan sähköisen liiketoiminnan ja asiakaspalvelutoiminnan työvälineenä käytettävän ekstranet–verkon sovellus- ja hyötypotentiaalia teollisuusyrityksissä. Lisäksi selvityksen kohteena ovat ekstranet –palvelu-kanavien strateginen suunnittelu ja kehitys yrityksissä sekä niiden liike-toiminnalliset vaikutukset. Tarkastelun kohteena on esimerkkiyrityksen ekstranet–palvelujärjestelmä ja siinä toimivat sovellukset toiminta-ominaisuuksineen. Lähitarkastelussa on yrityksen yhden ekstranet –moduulin konseptin kehittäminen, joka liittyy uuden sovelluksen pilottikehitys-hankkeeseen, jossa asiakkaalle on tarkoitus tarjota teknistä tukea ns. etädiagnostiikan avulla. Etäseurannan kohteena tässä moduulissa ovat asiakkaan liimausprosessin parametrit, jotka kuvaavat toimittajan liima-aineiden käyttäytymistä asiakkaan prosessissa. Parametritietojen siirtämisen relevanttiutta sekä asiakkaan prosessin problematiikkaa on selvitetty haastattelemalla yrityksen tuotekehityksen ja teknisen asiakaspalvelun avainhenkilöitä. Välittämällä asiakkaan prosessia koskevia tietoja ekstranetiin ja hyväksikäyttämällä niitä päästään merkittäviin molemminpuolisiin tehokkuus- ja kilpailuetuihin sekä kasvavaan asiakasuskollisuuteen. Työn tuloksena on esitetty kehitettävän sovelluksen konsepti, joka voi määritellä ennakkotiedot sitä rakentavalle ASP-toimittajalle sekä toimia vakiomallina sovelluksen käyttökohteita laajennettaessa.

Teollisuusjärjestelmien olosuhteiden hallinnan diagnostiikka

Tässä työssä tutkitaan kartonkikone- ja arkkisalin ilmastointijärjestelmän vaikutusta kartonkitehtaan olosuhteisiin. Ilmastoinnin ensisijainen tehtävä on taata prosessin häiriötön toiminta. Lisäksi hyvin toimivan ilmastointijärjestelmän avulla luodaan paremmat työskentelyolosuhteet sekä parannetaan rakennuksen toimintaolosuhteita. Kartonkitehtaan nykyiset olosuhteet selvitetään mittausten ja laskelmien avulla. Ilmastointijärjestelmän nykykuntoa ja tehokkuutta kartoitetaan havaintojen, mittausten ja laskelmien avulla. Olosuhteiden ja ilmastoinnin nykytilan kartoituksen avulla luodaan parannusehdotukset ilmastoinnin uusinnalle. Ilmastointijärjestelmän parannusehdotusten avulla hallitaan tuotannonlisäyksen aiheuttamat lisäkuormitukset sekä parannetaan työskentelyolosuhteita niin kesä- kuin talviolosuhteissa.

Konversio OBD:n ja sarjaliikenneportin välillä

Tämä insinöörityö on tehty Helsingin ammattikorkeakoululle osaksi suurempaa kokonaisuutta, jonka lopullisena tarkoituksena on lähettää auton OBD-diagnostiikkatiedot langattomasti paikkaan, jossa niitä voidaan analysoida, esimerkiksi autohuoltamolle. OBD2 (On-Board-Diagnostics II) on autoissa toimiva diagnostiikkajärjestelmä, joka tutkii auton eri osa-alueita, tallentaa tietoja näistä alueista ja ilmoittaa, jos jokin on vialla. Järjestelmä on nykyään laajalti käytössä. Ensimmäiset toteutukset OBD:stä nähtiin 80-luvun lopulla Yhdysvalloissa. Sitä seuraava toteutus OBD2 otettiin käyttöön 1996. EODB (European On Board Diagnostics) otettiin käyttöön Euroopassa vuonna 2001. Se on käytännössä sama kuin OBD2, mutta siihen on tehty muutamia muutoksia, jotka tekevät siitä paremmin sopivan Eurooppaan. Tässä insinöörityössä tutkitaan, kuinka tapahtuu konversio OBD-rajapinnan ja sarjaliikenneportin välillä. Tätä konversiota valittiin suorittamaan edullinen AGV2055-piirisarjaan perustuva OBD2-adapterin rakennussarja, joka kytketään tietokoneen sarjaliikenneporttiin. Tässä työssä perehdytään adapterin kokoamiseen ja tutustutaan sen toimintaan. Lisäksi kokeillaan kolmea eri ohjelmaa (yksi avoimen lähdekoodin ja kaksi kaupallista) suoritta-maan tulkkaamista ajoneuvon ja tietokoneen välillä.

Tuotekeskeisen palvelun prosessien kehittäminen teollisuusyrityksessä

Perinteisen valmistavan teollisuuden yritykset ovat pyrkineet kehittämään palveluja perinteisen tuotevalikoimansa rinnalle. Tuotekeskeinen palvelu on valmistettavaan laitteeseen liitettävä lisäpalvelu, jonka toimitus mahdollistuu siihen liitettyyn laitteeseen lisättyjen älykkäiden ominaisuuksien, kuten etädiagnostiikkalaitteiden avulla. Systemaattisesti kehitetyt prosessit ovat avainasemassa palvelun sujuvalle tuottamiselle. Tämän työn tavoitteena oli selvittää miten kohdeyrityksen tuotekeskeisen palvelun tuotantoon liittyvät prosessit voidaan integroida yrityksen muihin olemassa oleviin prosesseihin. Työn teoreettisessa osuudessa aihepiiriä lähestyttiin prosessijohtamisen ja siihen liittyvän termistön ja viitekehysten kautta, sekä lisäksi tutkittiin tuotekeskeisen palvelun käsitteistöä ja tähän kokonaisuuteen liittyviä prosesseja ja niiden kehittämistä. Teoreettista viitekehystä sovellettiin työn empiirisessä osassa kuvaamalla prosessien nykytilaa, sekä niihin liittyviä ongelmia ja kehityskohteita mm. haastattelujen ja olemassa olevan dokumentaation avulla. Nykytilan selvittäminen toimi lähtökohtana työn aikana käynnistetylle kehitysprojektille, jossa pyrittiin tuomaan tuotannollinen ja myynnillinen valmius valittuihin trukkimalleihin. Työssä syntyi toimintamalli niille tehtäville, joita yrityksessä täytyy toteuttaa tavoitteisiin pääsemiseksi, sekä laadittiin kuvaukset myynti- ja jälkiasennusprosesseihin.

”Kyllähän nyt pitäs olla jo semmonen aika, että pääsis niin kö keskustelemhan näistä asioista.” Tutkimus lappilaisten sotavankien, partisaanien uhrien ja huutolaisten elämänkulusta, voimavaroista, terveydestä ja sairauksista

Research into the course of life, mental stamina and health status of wartime prisoners, victims of Soviet partisan attacks, and paupers in Finnish Lapland The basis of this research comprised the issues raised during the interviews conducted in my work as a general practitioner in Lapland, regarding factors that have possibly affected the life stories and health conditions of Lappish people who had lived through the war as war prisoners, victims of partisan attacks, or paupers. The purpose of the study was to describe how the different life phases and experiences emerged from the interviewees’ stories and to identify their mental stamina. Another goal was to make observations on their health status, in which the main emphasis became to address mental symptoms. The cohort consisted of elderly Finns who lived in Lapland during the war and experienced war imprisonment, pauperism, or became victims of partisan attacks. All three groups consisted of 12 interviewees. The interview transcripts were read several times and then investigated using the content analysis methods applicable to the material. The research methodology was based on building awareness and understanding. Thematic tagging and data coding were used as structured analysis tools. In all three groups most of the interviewees clearly identified their mental stamina, the most fundamental of which were home, family and work. The war prisoners’ injuries and nervous sensibility symptoms had been shown in earlier studies on war prisoners, and on this basis they had been granted disability pensions. However, many of them had suppressed their traumatic experiences and mental difficulties, and they could not talk about these issues until at the time of these interviews held at old age. Four of them still suffered from a post-traumatic stress disorder. The victims of Soviet partisans had had to carry their mental load alone for decades before the cruel ravages on civilians in remote areas of Lapland became publicly known. Most of them still had disturbing nervous sensibility symptoms. Four interviewees had a post-traumatic stress disorder, and in addition to these, the mental symptoms of one had developed into a post-traumatic stress disorder during old age. Many of the interviewees who had been left paupers remembered their childhood as filled with grief and feelings of inferiority, and had nightmares relating to their wartime experiences. Yet none of them suffered from post-traumatic stress disorder. The results showed that the exceptional suffering caused by the war, the wartime imprisonment and the devastating attacks by Soviet partisans had led especially to mental difficulties. These were left almost completely unnoticed in the post-war conditions, and the war victims were unable to seek help on their own. Based on the results, our health care for the elderly should focus on familiarization with the individual experiences and life stories of each elderly person. This can facilitate geriatric diagnostics and individual therapy planning. Empathic familiarization with the life experiences of the elderly may strengthen their mental stamina and improve the quality of successful aging.

Bile duct stones, strictures and iatrogenic lesions: studies on early diagnosis and treatment.

Aims: This study was carried out to investigate the role of common liver function tests, and the degree of common bile duct dilatation in the differential diagnosis of extrahepatic cholestasis, as well as the occurrence, diagnosis and treatment of iatrogenic bile duct injuries. In bile duct injuries, special attention was paid to gender and severity distribution and long-term results. Patients and methods: All consecutive patients with diagnosed common bile duct stones or malignant strictures in ERCP between August 2000 and November 2003. Common liver function tests were measured in the morning before ERCP on all of these 212 patients, and their common bile duct diameter was measured from ERCP films. Between January 1995 and April 2002, 3736 laparoscopic cholecystectomies were performed and a total of 32 bile duct injuries were diagnosed. All pre-, per-, and postoperative data were collected retrospectively; and the patients were also interviewed by phone. Results: Plasma bilirubin proved to be the best discriminator between CBD stones and malignant strictures (p≤0.001 compared to other liver function tests and degree of common bile duct dilatation). The same effect was seen in Receiver Operating Characteristics curves (AUC 0.867). With a plasma bilirubin cut-off value of 145 μmol/l, four out of five patients could be classified correctly. The degree of common bile duct dilatation proved to be worthless in differential diagnostics. After laparoscopic cholecystectomy the total risk for bile duct injury was 0.86%, including cystic duct leaks. 86% of severe injuries and 88% of injuries requiring operative treatment were diagnosed in females. All the cystic duct leakages and 87% of the strictures were treated endoscopically. Good long-term results were seen in 84% of the whole study population. Conclusions: Plasma bilirubin is the most effective liver function test in differential diagnosis between CBD stones and malignant strictures. The only value of common bile duct dilatation is its ability to verify the presence of extrahepatic cholestasis. Female gender was associated with higher number of iatrogenic bile duct injuries, and in particular, most of the major complications occur in females. Most of the cystic duct leaks and common bile duct strictures can be treated endoscopically. The long-term results in our institution are at an internationally acceptable level.

Diagnostiikkayksikön kehittäminen rautatiesovellukseen

Suomessa rautatiellä käytetään muutamia tuhansia sähköisesti ohjattavia vaihteita, joiden toiminta varmistetaan määräajoin tehtävillä huolloilla. Työssä on tutkittu, onko kehitteillä olevalla vaihteenohjaimella mahdollista mitata ja analysoida vaihteen kunnonvalvontaan liittyviä suureita. Tutkimuksessa on tutustuttu rautatietekniikkaan sekä vaihdetta ohjaavaan vaihteenkääntölaitteeseen. Tutkimuksen perusteella on kehitetty sulautettuun järjestelmään perustuva mittaus- ja analysointijärjestelmäsovellus, joka tekee mittauksia, havaintoja ja johtopäätöksiä vaihteen toiminnasta. Mittaukset on suoritettu moottoria ohjaavista johtimista ja mittaustulokset on tallennettu analysointia ja jatkokäsittelyä varten. Ohjelmiston toimintaa on varmistettu mittauksilla sekä laboratorio-olosuhteissa että oikealla vaihteella testausympäristössä. Saatujen tulosten perusteella voidaan todeta, että kehitetyllä laitteistolla on mahdollista toteuttaa vaihdetta diagnosoivia mittauksia riittävän laadukkaasti ja tarkasti. Työssä kehitetty järjestelmäalusta soveltuu käytettäväksi vaihteen diagnostiikan jatkokehittämiselle.

Early Detection of Alzheimer's Disease Beta-amyloid Pathology -Applicability of Positron Emission Tomography with the Amyloid Radioligand 11C-PIB

Early Detection of Alzheimer's Disease Beta-amyloid Pathology -Applicability of Positron Emission Tomography with the Amyloid Radioligand ¹¹C-PIB Accumulation of beta amyloid (Abeta) in the brain is characteristic for Alzheimer’s disease (AD). Carbon-11 labeled 2-(4’-methylaminophenyl)-6-hydroxybenzothiazole (¹¹C-PIB) is a novel positron emission tomography (PET) amyloid imaging agent that appears to be applicable for in vivo Abeta plaque detection and quantitation. The biodistribution and radiation dosimetry of ¹¹C-PIB were investigated in 16 healthy subjects. The reproducibility of a simplified ¹¹C-PIB quantitation method was evaluated with a test-retest study on 6 AD patients and 4 healthy control subjects. Brain ¹¹C-PIB uptake and its possible association with brain atrophy rates were studied over a two-year follow-up in 14 AD patients and 13 healthy controls. Nine monozygotic and 8 dizygotic twin pairs discordant for cognitive impairment and 9 unrelated controls were examined to determine whether brain Abeta accumulation could be detected with ¹¹C-PIB PET in cognitively intact persons who are at increased genetic risk for AD. The highest absorbed radiation dose was received by the gallbladder wall (41.5 mjuGy/MBq). About 20 % of the injected radioactivity was excreted into urine, and the effective whole-body radiation dose was 4.7 mjuSv/MBq. Such a dose allows repeated scans of individual subjects. The reproducibility of the simplified ¹¹C-PIB quantitation was good or excellent both at the regional level (VAR 0.9-5.5 %) and at the voxel level (VAR 4.2-6.4 %). ¹¹C-PIB uptake did not increase during 24 months’ follow-up of subjects with mild or moderate AD, even though brain atrophy and cognitive decline progressed. Baseline neocortical ¹¹C-PIB uptake predicted subsequent volumetric brain changes in healthy control subjects (r = 0.725, p = 0.005). Cognitively intact monozygotic co-twins – but not dizygotic co-twins – of memory-impaired subjects exhibited increased ¹¹C-PIB uptake (117-121 % of control mean) in their temporal and parietal cortices and the posterior cingulate (p<0.05), when compared with unrelated controls. This increased uptake may be representative of an early AD process, and genetic factors seem to play an important role in the development of AD-like Abeta plaque pathology. ¹¹C-PIB PET may be a useful method for patient selection and follow-up for early-phase intervention trials of novel therapeutic agents. AD might be detectable in high-risk individuals in its presymptomatic stage with ¹¹C-PIB PET, which would have important consequences both for future diagnostics and for research on disease-modifying treatments.

Evolution of Bioaffinity Reagents by Phage Display

Antibodies are natural binding proteins produced in vertebrates as a response to invading pathogens and foreign substances. Because of their capability for tight and specific binding, antibodies have found use as binding reagents in research and diagnostics. Properties of cloned recombinant antibodies can be further improved by means of in vitro evolution, combining mutagenesis with subsequent phage display selection. It is also possible to isolate entirely new antibodies from vast naïve or synthetic antibody libraries by phage display. In this study, library techniques and phage display selection were applied in order to optimise binding scaffolds and antigen recognition of antibodies, and to evolve new and improved bioaffinity reagents. Antibody libraries were generated by random and targeted mutagenesis. Expression and stability were mainly optimised by the random methods whereas targeted randomisation of the binding site residues was used for optimising the binding properties. Trinucleotide mutagenesis allowed design of defined randomisation patterns for a synthetic antibody library. Improved clones were selected by phage display. Capture by a specific anti- DHPS antibody was exploited in the selection of improved phage display of DHPS. Efficient selection for stability was established by combining phage display selection with denaturation under reducing conditions. Broad-specific binding of a generic anti-sulfonamide antibody was improved by selection with one of the weakest binding sulfonamides. In addition, p9 based phage display was studied in affinity selection from the synthetic library. A TIM barrel protein DHPS was engineered for efficient phage display by combining cysteinereplacement with random mutagenesis. The resulting clone allows use of phage display in further engineering of DHPS and possibly use as an alternative-binding scaffold. An anti-TSH scFv fragment, cloned from a monoclonal antibody, was engineered for improved stability to better suite an immunoassay. The improved scFv tolerates 8 – 9 °C higher temperature than the parental scFv and should have sufficient stability to be used in an immunoanalyser with incubation at 36 °C. The anti-TSH scFv fragment was compared with the corresponding Fab fragment and the parental monoclonal antibody as a capturing reagent in a rapid 5-min immunoassay for TSH. The scFv fragment provided some benefits over the conventionally used Mab in anayte-binding capacity and assay kinetics. However, the recombinant Fab fragment, which had similar kinetics to the scFv, provided a more sensitive and reliable assay than the scFv. Another cloned scFv fragment was engineered in order to improve broad-specific recognition of sulfonamides. The improved antibody detects different sulfonamides at concentrations below the maximum residue limit (100 μg/kg in EU and USA) and allows simultaneous screening of different sulfonamide drug residues. Finally, a synthetic antibody library was constructed and new antibodies were generated and affinity matured entirely in vitro. These results illuminate the possibilities of phage display and antibody engineering for generation and optimisation of binding reagents in vitro and indicate the potential of recombinant antibodies as affinity reagents in immunoassays.

Studies on 68Ga-Based Agents for PET Imaging of Cancer and Inflammation

Studies on ⁶⁸Ga-Based Agents for PET Imaging of Cancer and Inflammation Positron emission tomography (PET) is based on the use of radiolabeled agents and facilitates in vivo imaging of biological processes, such as cancer. Because the detection of cancer is demanding and is often obscured by inflammation, there is a demand for better PET imaging agents. The aim was to preliminarily evaluate new PET agents for imaging cancer and inflammation using experimental models. ⁶⁸Ga-chloride and peptides, ⁶⁸Ga-labeled through 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), targeting matrix metalloproteinase-9 (MMP-9) were tested for tumor imaging. In addition, a ⁶⁸Ga-DOTA-conjugated peptide targeting vascular adhesion protein-1 (VAP-1), was tested for inflammation imaging. The ⁶⁸Ga-based imaging agents described here showed potential features by passing the essential in vitro tests, proceeding further to preclinical in vivo evaluation and being able to visualize the target. The target uptake and target-to-background ratios of ⁶⁸Ga-based agents were, however, not optimal. ⁶⁸Ga-chloride showed slow clearance caused by its binding to blood transferrin. In the case of ⁶⁸Ga-DOTA-peptides low in vivo stability and/or low lipophilicity led to too rapid blood clearance and urinary excretion. The properties of ⁶⁸Ga-labeled peptides are modifiable, as shown with matrix metalloproteinase-9 targeting ligands. In the conclusion of this PhD thesis, ⁶⁸Ga-based agents for PET imaging of cancer and inflammation could be applied in the development of drugs, earlier diagnostics and following-up of the efficacy of therapies.

Monitoring of centrifugal pump operation by a frequency converter

Centrifugal pumps are widely used in industrial and municipal applications, and they are an important end-use application of electric energy. However, in many cases centrifugal pumps operate with a significantly lower energy efficiency than they actually could, which typically has an increasing effect on the pump energy consumption and the resulting energy costs. Typical reasons for this are the incorrect dimensioning of the pumping system components and inefficiency of the applied pump control method. Besides the increase in energy costs, an inefficient operation may increase the risk of a pump failure and thereby the maintenance costs. In the worst case, a pump failure may lead to a process shutdown accruing additional costs. Nowadays, centrifugal pumps are often controlled by adjusting their rotational speed, which affects the resulting flow rate and output pressure of the pumped fluid. Typically, the speed control is realised with a frequency converter that allows the control of the rotational speed of an induction motor. Since a frequency converter can estimate the motor rotational speed and shaft torque without external measurement sensors on the motor shaft, it also allows the development and use of sensorless methods for the estimation of the pump operation. Still today, the monitoring of pump operation is based on additional measurements and visual check-ups, which may not be applicable to determine the energy efficiency of the pump operation. This doctoral thesis concentrates on the methods that allow the use of a frequency converter as a monitoring and analysis device for a centrifugal pump. Firstly, the determination of energy-efficiency- and reliability-based limits for the recommendable operating region of a variable-speed-driven centrifugal pump is discussed with a case study for the laboratory pumping system. Then, three model-based estimation methods for the pump operating location are studied, and their accuracy is determined by laboratory tests. In addition, a novel method to detect the occurrence of cavitation or flow recirculation in a centrifugal pump by a frequency converter is introduced. Its sensitivity compared with known cavitation detection methods is evaluated, and its applicability is verified by laboratory measurements for three different pumps and by using two different frequency converters. The main focus of this thesis is on the radial flow end-suction centrifugal pumps, but the studied methods can also be feasible with mixed and axial flow centrifugal pumps, if allowed by their characteristics.

Defects in Persistent Luminescence Materials

Persistent luminescence materials can store energy from solar radiation or artificial lighting and release it over a period of several hours without a continuous excitation source. These materials are widely used to improve human safety in emergency and traffic signalization. They can also be utilized in novel applications including solar cells, medical diagnostics, radiation detectors and structural damage sensors. The development of these materials is currently based on methods based on trial and error. The tailoring of new materials is also hindered by the lack of knowledge on the role of their intrinsic and extrinsic lattice defects in the appropriate mechanisms. The goal of this work was to clarify the persistent luminescence mechanisms by combining ab initio density functional theory (DFT) calculations with selected experimental methods. The DFT approach enables a full control of both the nature of the defects and their locations in the host lattice. The materials studied in the present work, the distrontium magnesium disilicate (Sr₂MgSi₂O₇) and strontium aluminate (SrAl₂O₄) are among the most efficient persistent luminescence hosts when doped with divalent europium Eu²⁺ and co-doped with trivalent rare earth ions R³⁺ (R: Y, La-Nd, Sm, Gd-Lu). The polycrystalline materials were prepared with the solid state method and their structural and phase purity was confirmed by X-ray powder diffraction. Their local crystal structure was studied by high-resolution transmission electron microscopy. The crystal and electronic structure of the nondoped as well as Eu²⁺, R^2+/3+ and other defect containing materials were studied using DFT calculations. The experimental trap depths were obtained using thermoluminescence (TL) spectroscopy. The emission and excitation of Sr₂MgSi₂O₇:Eu²+,Dy³⁺ were also studied. Significant modifications in the local crystal structure due to the Eu²⁺ ion and lattice defects were found by the experimental and DFT methods. The charge compensation effects induced by the R³⁺ co-doping further increased the number of defects and distortions in the host lattice. As for the electronic structure of Sr₂MgSi₂O₇ and SrAl₂O₄, the experimental band gap energy of the host materials was well reproduced by the calculations. The DFT calculated Eu²⁺ and R^2+/3+ 4fn as well as 4f^n-15d¹ ground states in the Sr₂MgSi₂O₇ band structure provide an independent verification for an empirical model which is constructed using rather sparse experimental data for the R³⁺ and especially the R²⁺ ions. The intrinsic and defect induced electron traps were found to act together as energy storage sites contributing to the materials’ efficient persistent luminescence. The calculated trap energy range agreed with the trap structure of Sr₂MgSi₂O₇ obtained using TL measurements. More experimental studies should be carried out for SrAl₂O₄ to compare with the DFT calculations. The calculated and experimental results show that the electron traps created by both the rare earth ions and vacancies are modified due to the defect aggregation and charge compensation effects. The relationships between this modification and the energy storage properties of the solid state materials are discussed.

Luminescence-based assay methods in drug discovery

The drug discovery process is facing new challenges in the evaluation process of the lead compounds as the number of new compounds synthesized is increasing. The potentiality of test compounds is most frequently assayed through the binding of the test compound to the target molecule or receptor, or measuring functional secondary effects caused by the test compound in the target model cells, tissues or organism. Modern homogeneous high-throughput-screening (HTS) assays for purified estrogen receptors (ER) utilize various luminescence based detection methods. Fluorescence polarization (FP) is a standard method for ER ligand binding assay. It was used to demonstrate the performance of two-photon excitation of fluorescence (TPFE) vs. the conventional one-photon excitation method. As result, the TPFE method showed improved dynamics and was found to be comparable with the conventional method. It also held potential for efficient miniaturization. Other luminescence based ER assays utilize energy transfer from a long-lifetime luminescent label e.g. lanthanide chelates (Eu, Tb) to a prompt luminescent label, the signal being read in a time-resolved mode. As an alternative to this method, a new single-label (Eu) time-resolved detection method was developed, based on the quenching of the label by a soluble quencher molecule when displaced from the receptor to the solution phase by an unlabeled competing ligand. The new method was paralleled with the standard FP method. It was shown to yield comparable results with the FP method and found to hold a significantly higher signal-tobackground ratio than FP. Cell-based functional assays for determining the extent of cell surface adhesion molecule (CAM) expression combined with microscopy analysis of the target molecules would provide improved information content, compared to an expression level assay alone. In this work, immune response was simulated by exposing endothelial cells to cytokine stimulation and the resulting increase in the level of adhesion molecule expression was analyzed on fixed cells by means of immunocytochemistry utilizing specific long-lifetime luminophore labeled antibodies against chosen adhesion molecules. Results showed that the method was capable of use in amulti-parametric assay for protein expression levels of several CAMs simultaneously, combined with analysis of the cellular localization of the chosen adhesion molecules through time-resolved luminescence microscopy inspection.

Transcriptional Profiling of Organ‐Specific Autoimmunity in Human

Our understanding of the pathogenesis of organ‐specific autoinflammation has been restricted by limited access to the target organs. Peripheral blood, however, as a preferred transportation route for immune cells, provides a window to assess the entire immune system throughout the body. Transcriptional profiling with RNA stabilizing blood collection tubes reflects in vivo expression profiles at the time the blood is drawn, allowing detection of the disease activity in different samples or within the same sample over time. The main objective of this Ph.D. study was to apply gene‐expression microarrays in the characterization of peripheral blood transcriptional profiles in patients with autoimmune diseases. To achieve this goal a custom cDNA microarray targeted for gene‐expression profiling of human immune system was designed and produced. Sample collection and preparation was then optimized to allow gene‐expression profiling from whole‐blood samples. To overcome challenges resulting from minute amounts of sample material, RNA amplification was successfully applied to study pregnancy related immunosuppression in patients with multiple sclerosis (MS). Furthermore, similar sample preparation was applied to characterize longitudinal genome‐wide expression profiles in children with type 1 diabetes (T1D) associated autoantibodies and eventually clinical T1D. Blood transcriptome analyses, using both the ImmunoChip cDNA microarray with targeted probe selection and genome‐wide Affymetrix U133 Plus 2.0 oligonucleotide array, enabled monitoring of autoimmune activity. Novel disease related genes and general autoimmune signatures were identified. Notably, down‐regulation of the HLA class Ib molecules in peripheral blood was associated with disease activity in both MS and T1D. Taken together, these studies demonstrate the potential of peripheral blood transcriptional profiling in biomedical research and diagnostics. Imbalances in peripheral blood transcriptional activity may reveal dynamic changes that are relevant for the disease but might be completely missed in conventional cross‐sectional studies.

Broadband excitation in the system identification of active magnetic bearing rotor systems

One of the targets of the climate and energy package of the European Union is to increase the energy efficiency in order to achieve a 20 percent reduction in primary energy use compared with the projected level by 2020. The energy efficiency can be improved for example by increasing the rotational speed of large electrical drives, because this enables the elimination of gearboxes leading to a compact design with lower losses. The rotational speeds of traditional bearings, such as roller bearings, are limited by mechanical friction. Active magnetic bearings (AMBs), on the other hand, allow very high rotational speeds. Consequently, their use in large medium- and high-speed machines has rapidly increased. An active magnetic bearing rotor system is an inherently unstable, nonlinear multiple-input, multiple-output system. Model-based controller design of AMBs requires an accurate system model. Finite element modeling (FEM) together with the experimental modal analysis provides a very accurate model for the rotor, and a linearized model of the magneticactuators has proven to work well in normal conditions. However, the overall system may suffer from unmodeled dynamics, such as dynamics of foundation or shrink fits. This dynamics can be modeled by system identification. System identification can also be used for on-line diagnostics. In this study, broadband excitation signals are adopted to the identification of an active magnetic bearing rotor system. The broadband excitation enables faster frequency response function measurements when compared with the widely used stepped sine and swept sine excitations. Different broadband excitations are reviewed, and the random phase multisine excitation is chosen for further study. The measurement times using the multisine excitation and the stepped sine excitation are compared. An excitation signal design with an analysis of the harmonics produced by the nonlinear system is presented. The suitability of different frequency response function estimators for an AMB rotor system are also compared. Additionally, analytical modeling of an AMB rotor system, obtaining a parametric model from the nonparametric frequency response functions, and model updating are discussed in brief, as they are key elements in the modeling for a control design. Theoretical methods are tested with a laboratory test rig. The results conclude that an appropriately designed random phase multisine excitation is suitable for the identification of AMB rotor systems.