Vitamin B12 Deficiency in the aged: Laboratory Diagnosis, Prevalence and Clinical Profile

B₁₂-vitamiinin puute iäkkäillä: laboratoriodiagnostiikka, yleisyys ja yhteys sairastavuuteen Tausta: B12-vitamiinin puute on yleistä iäkkäillä ja se tulisi todeta riittävän varhaisessa vaiheessa palautumattomien vaurioiden estämiseksi. On epäselvää pitäisikö diagnostiikka kohdistaa tiettyihin riskiryhmiin vai mahdollisesti seuloa valikoimatonta vanhusväestöä. Myöskään yksimielisyyttä laboratoriotutkimusten valinnasta ei ole. Tavoitteet: Tutkimuksen tarkoituksena oli evaluoida uutta HoloTC RIA menetelmää ja tuottaa viitearvot sille, selvittää B12-vitamiinin puutteen yleisyys, yhteys sairastavuuteen ja mahdolliset riskitekijät suomalaisessa vanhusväestössä, arvioida munuaisfunktion vaikusta B12-vitamiinin puutteen laboratoriotutkimuksiin ja näiden perusteella ehdottaa suomalaiseen terveydenhuoltoon sopivaa laboratoriotutkimusstrategiaa. Aineisto ja menetelmät: Liedon iäkkäät -tutkimuksen vanhusaineisto on edustava otos yhden kunnan yli 65-vuotiaasta väestöstä, yhteensä 1260 henkilöä. Tutkittavat kävivät lääkärintarkastuksessa, ja heistä on käytettävissä runsaasti laboratoriotutkimuksia sekä tiedot sairauksista, ruokavaliosta, lääkkeiden ja vitamiinivalmisteiden käytöstä, dementiaseula ja depressiokysely. Viitearvoaineistoa varten kerättiin näytteet 84 vapaaehtoisesta terveestä aikuisesta ja menetelmäevaluaatiota varten 107 sairaalapotilaasta. Tulokset: HoloTC RIA menetelmän toistettavuus oli hyvä manuaalimenetelmäksi. 95%:n viiteväli holotranskobalamiinille oli 37-171 pmol/l. Kaikilla tutkittavilla, joilla oli muilla laboratoriotutkimuksilla osoitettu todennäköinen B12-vitamiinin puute, myös holotranskobalamiini oli viitealueen alarajaa pienempi. Suurentuneella kystatiini C-pitoisuudella osoitettu munuaisten vajaatoiminta korreloi voimakkaasti homokysteiinin (rs=0.53, p<0.001) ja metyylimalonihapon (rs=0.27, p<0.001) pitoisuuksiin, mutta ei kokonais-B12-vitamiinin (rs=- 0.04, p=0.227) tai holotranskobamiinin (rs=-0.01, p=0.817) pitoisuuksiin. Suomalaisessa vanhusväestössä B12-vitamiinin puutteen prevalenssi oli 12%. Kokonais- B12-vitamiinin pitoisuus oli matala (<150 pmol/l) 6%:lla. Miessukupuoli (OR 1.9, 95% CI 1.2-2.9), ikä ≥75 (OR 2.2, 95% CI 1.4-3.4) ja maitotuotteiden välttäminen (OR 2.3, 95% CI 1.2-4.4) lisäsivät B12-vitamiinin puutteen riskiä, mutta anemia (OR 1.3, 95% CI 0.7-2.3) tai makrosytoosi (OR 1.2, 95% CI 0.6-2.7) eivät. Päätelmät: Diagnosoimaton B12-vitamiinin puute on yleistä iäkkäillä, mutta kliinisesti merkityksellistä spesifistä riskiryhmää ei löydy. Koska anemian ja makrosytoosin puuttuminen ei poissulje B12-vitamiinin puutetta ja munuaisten vajaatoiminta heikentää metabolisten merkkiaineiden käyttökelpoisuutta, kokonais-B12-vitamiinia suositellaan ensisijaiseksi laboratoriotutkimukseksi epäiltäessä B12-vitamiinin puutetta ja tarvittaessa varmentavina tutkimuksina käytetään homokysteiiniä ja holotranskobalamiinia.

Effect of physico-chemical conditions and operating parameters on flux and retention of different components in ultrafiltration and nanofiltration fractionation of sweet whey

Tässä väitöstutkimuksessa tutkittiin fysikaaliskemiallisten olosuhteiden ja toimintaparametrien vaikutusta juustoheran fraktiointiin. Kirjallisuusosassa on käsitelty heran ympäristövaikutusta, heran hyödyntämistä ja heran käsittelyä kalvotekniikalla. Kokeellinen osa on jaettu kahteen osaan, joista ensimmäinen käsittelee ultrasuodatusta ja toinen nanosuodatusta juustoheran fraktioinnissa. Ultrasuodatuskalvon valinta tehtiin perustuen kalvon cut-off lukuun, joka oli määritetty polyetyleeniglykoliliuoksilla olosuhteissa, joissa konsentraatiopolariosaatioei häiritse mittausta. Kriittisen vuon konseptia käytettiin sopivan proteiinikonsentraation löytämiseksi ultrasuodatuskokeisiin, koska heraproteiinit ovat tunnetusti kalvoa likaavia aineita. Ultrasuodatuskokeissa tutkittiin heran eri komponenttien suodattumista kalvon läpi ja siihen vaikuttavia ominaisuuksia. Herapermeaattien peptidifraktiot analysoitiin kokoekskluusiokromatografialla ja MALDI-TOF massaspektrometrillä. Kokeissa käytettävien nanosuodatuskalvojen keskimääräinen huokoskoko analysoitiin neutraaleilla liukoisilla aineilla ja zeta-potentiaalit virtauspotentiaalimittauksilla. Aminohappoja käytettiin malliaineina tutkittaessa huokoskoon ja varauksen merkitystä erotuksessa. Aminohappojen retentioon vaikuttivat pH ja liuoksen ionivahvuus sekä molekyylien väliset vuorovaikutukset. Heran ultrasuodatuksessa tuotettu permeaatti, joka sisälsi pieniä peptidejä, laktoosia ja suoloja, nanosuodatettiin happamassa ja emäksisessä pH:ssa. Emäksisissä oloissa tehdyssä nanosuodatuksessa foulaantumista tapahtui vähemmän ja permeaattivuo oli parempi. Emäksisissä oloissa myös selektiivisyys laktoosin erotuksessa peptideistä oli parempi verrattuna selektiivisyyteen happamissa oloissa.

Critical flux and fouling in ultrafiltration of proteins

In this thesis different parameters influencing critical flux in protein ultrafiltration and membrane foul-ing were studied. Short reviews of proteins, cross-flow ultrafiltration, flux decline and criticalflux and the basic theory of Partial Least Square analysis (PLS) are given at the beginning. The experiments were mainly performed using dilute solutions of globular proteins, commercial polymeric membranes and laboratory scale apparatuses. Fouling was studied by flux, streaming potential and FTIR-ATR measurements. Critical flux was evaluated by different kinds of stepwise procedures and by both con-stant pressure and constant flux methods. The critical flux was affected by transmembrane pressure, flow velocity, protein concentration, mem-brane hydrophobicity and protein and membrane charges. Generally, the lowest critical fluxes were obtained at the isoelectric points of the protein and the highest in the presence of electrostatic repulsion between the membrane surface and the protein molecules. In the laminar flow regime the critical flux increased with flow velocity, but not any more above this region. An increase in concentration de-creased the critical flux. Hydrophobic membranes showed fouling in all charge conditionsand, furthermore, especially at the beginning of the experiment even at very low transmembrane pressures. Fouling of these membranes was thought to be due to protein adsorption by hydrophobic interactions. The hydrophilic membranes used suffered more from reversible fouling and concentration polarisation than from irreversible foul-ing. They became fouled at higher transmembrane pressures becauseof pore blocking. In this thesis some new aspects on critical flux are presented that are important for ultrafiltration and fractionation of proteins.

The central histaminergic neurons in vitro, and their neuroprotective role in excitotoxic cell death in the postnatal rat hippocampus.

Histamine acts as a neurotransmitter in the central nervous system. Brain histamine in synthesized in neurons located to the posterior hypothalamus, from where these neurons send their projections to different parts of the brain. Released histamine participates in the regulation of several physiological functions such as arousal, attention and body homeostasis. Disturbances in the histaminergic system have been detected in diseases such as epilepsy, sleep disorders, anxiety, depression, Alzheimer’s disease, and schizophrenia. The purpose of this thesis was to develop optimal culture conditions for the histaminergic neurons, to study their detailed morphology, and to find out their significance in the kainic acid (KA)-induced neuronal death in the immature rat hippocampus. The morphology of the histaminergic neurons in vitro was comparable with the earlier findings. Histamine-containing vesicles were found in the axon but also in the cell body and dendrites suggesting a possibility for the somatodendritic release. Moreover, histamine was shown to be colocalized with the vesicular monoamine transporter 2 (VMAT2) suggesting that VMAT2 transports histamine to the subcellular storage vesicles. Furthermore, histamine was localized with γ-aminobutyric acid (GABA) in distinct storage vesicles and with neuropeptide galanin partly in the same storage vesicles suggesting different corelease mechanisms for GABA and galanin with histamine. In the organotypic hippocampal slice cultures, KA-induced neuronal death was first detected 12 h after the treatment being restricted mainly to the CA3 subregion. Moreover, cell death was irreversible, since the 48 h recovery period did not save the cells, but instead increased the damage. Finally, neuronal death was suggested to be necrotic, since intracellular apoptotic pathways were not activated, and the morphological changes detected with the electron microscopy were characteristic for necrosis. In the coculture system of the hippocampal and posterior hypothalamic slices, histaminergic neurons significantly decreased epileptiform burst activity and neuronal death in the hippocampal slices, this effect being mediated by histamine 1 (H1) and 3 (H3) receptors. In conclusion, the histaminergic neurons were maintained succesfully in the in vitro conditions exhibiting comparable morphological characteristics as detected earlier in vivo. Moreover, they developed functional innervations within the hippocampal slices in the coculture system. Finally, the KA-induced regionspecific, irreversible and necrotic hippocampal pyramidal cell damage was significantly decreased by the histaminergic neurons through H1 and H3 receptors.

Reaction Kinetics and Viscosity Modelling in the Fusion Syntheses of Ca- and Ca/Mg-resinates

Rosin is a natural product from pine forests and it is used as a raw material in resinate syntheses. Resinates are polyvalent metal salts of rosin acids and especially Ca- and Ca/Mg- resinates find wide application in the printing ink industry. In this thesis, analytical methods were applied to increase general knowledge of resinate chemistry and the reaction kinetics was studied in order to model the non linear solution viscosity increase during resinate syntheses by the fusion method. Solution viscosity in toluene is an important quality factor for resinates to be used in printing inks. The concept of critical resinate concentration, c crit, was introduced to define an abrupt change in viscosity dependence on resinate concentration in the solution. The concept was then used to explain the non-inear solution viscosity increase during resinate syntheses. A semi empirical model with two estimated parameters was derived for the viscosity increase on the basis of apparent reaction kinetics. The model was used to control the viscosity and to predict the total reaction time of the resinate process. The kinetic data from the complex reaction media was obtained by acid value titration and by FTIR spectroscopic analyses using a conventional calibration method to measure the resinate concentration and the concentration of free rosin acids. A multivariate calibration method was successfully applied to make partial least square (PLS) models for monitoring acid value and solution viscosity in both mid-infrared (MIR) and near infrared (NIR) regions during the syntheses. The calibration models can be used for on line resinate process monitoring. In kinetic studies, two main reaction steps were observed during the syntheses. First a fast irreversible resination reaction occurs at 235 °C and then a slow thermal decarboxylation of rosin acids starts to take place at 265 °C. Rosin oil is formed during the decarboxylation reaction step causing significant mass loss as the rosin oil evaporates from the system while the viscosity increases to the target level. The mass balance of the syntheses was determined based on the resinate concentration increase during the decarboxylation reaction step. A mechanistic study of the decarboxylation reaction was based on the observation that resinate molecules are partly solvated by rosin acids during the syntheses. Different decarboxylation mechanisms were proposed for the free and solvating rosin acids. The deduced kinetic model supported the analytical data of the syntheses in a wide resinate concentration region, over a wide range of viscosity values and at different reaction temperatures. In addition, the application of the kinetic model to the modified resinate syntheses gave a good fit. A novel synthesis method with the addition of decarboxylated rosin (i.e. rosin oil) to the reaction mixture was introduced. The conversion of rosin acid to resinate was increased to the level necessary to obtain the target viscosity for the product at 235 °C. Due to a lower reaction temperature than in traditional fusion synthesis at 265 °C, thermal decarboxylation is avoided. As a consequence, the mass yield of the resinate syntheses can be increased from ca. 70% to almost 100% by recycling the added rosin oil.

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.

Jatkuvatoimisen natriumhypokloriittireaktorin mallinnus

Natriumhypokloriittia voidaan valmistaa kloorista ja lipeästä jatkuvatoimisessa absorberissa. Tässä työssä tutkittiin kokeellisesti, miten kaasun ja nesteen virtausnopeudet vaikuttavat täytekappalekolonnin tulvimiseen ja painehäviöön, kuinka nopeasti kloori absorboituu lipeään ja kuinka suuri hypokloriittiliuoksen kierrätys tarvitaan, ettei hypokloriitti ala hajota. Lisäksi luotiin matemaattinen malli, jolla voidaan mitoittaa jatkuvatoiminen vastavirtaperiaatteella toimiva natriumhypokloriittireaktori. Kloori–lipeäsysteemin havaittiin tulvivan suuremmilla virtausnopeuksilla kuin ilma–vesisysteemin. Tosin osa kloorista absorboituu jo ennen täytekappalekerrosta, minkä vuoksi kaasun todellinen virtausnopeus täytekappalekerroksen alaosassa on pienempi kuin mitattu arvo. Kolonnin painehäviö nousee erittäin jyrkästi tulvimispisteen läheisyydessä. Koska kloori absorboituu lähes täydellisesti ja vain kolonnin alaosa tulvii, voidaan kolonnia painehäviön kannalta operoida lähellä tulvimispistettä. Sekä mallinnuksen että koetulosten perusteella yli 99,99 % kloorista absorboituu koeolosuhteissa kahden metrin täytekappalekerroksessa. Nopea absorptio johtuu erittäin nopeasta, irreversiibelistä kloorin reaktiosta ja prosessille tyypillisestä natriumhydroksidikonversion rajoittamisesta alle 94 %:iin. Jotta varmistetaan, ettei hypokloriitti ala hajota, valmista hypokloriittiliuosta täytyy kierrättää kolonniin vähintään noin 4-kertainen määrä tuoreen lipeän syöttömäärän nähden.

Kainic acid-induced seizures: inflammation and excitotoxic neuronal damage in the developing rat hippocampus

Epileptic seizures are harmful to the developing brain. During epileptic seizures, overactivation of glutamate receptors (GluR) leads to neuronal degeneration, defined as excitotoxicity. The hippocampus is especially vulnerable to excitotoxic neuronal death, but its mechanism has remained incompletely known in the developing brain. Recently, signs of activation of inflammatory processes after epileptic seizures have been detected in the hippocampus. The purpose of this thesis was to study the inflammatory reaction and death mechanisms in excitoxic neurodegeneration induced by the glutamate analogue kainic acid (KA) in the developing hippocampus. Organotypic hippocampal slice cultures (OHCs), prepared from 6-7-day-old rats (P6-7) and treated with KA, served as an in vitro model. KA-induced status epilepticus in P9 and P21 rats was used as an in vivo model. The results showed that the pyramidal cell layers of the hippocampus were the most susceptible to irreversible and age-specific neurodegeneration, which occurred in the juvenile (P21), but not in the immature (P9), rat hippocampus. The primary death mechanism was necrosis as there were no significant changes in the expression of selected apoptosis markers and morphological cellular features of necrosis were found. Inflammatory response was similarly age-dependent after KA treatment as a rapid, fulminant and wide response was detected in the juvenile, but not in the immature, rat brain. An anti-inflammatory drug treatment, given before KA, was not neuroprotective in OHCs, possibly because of the timing of the treatment. In summary, the results suggest that KA induces an age-dependent inflammatory response and necrotic neurodegeneration, which may cause disturbances in hippocampal connectivity and promote epileptogenesis.

Ferromagnetic properties of pyrolytic nanocarbon

The work reported in this thesis is dedicated to irreversible magnetic properties in pyrolytic nanocarbon samples. Based on atomic force microscope images, the samples consist of carbon clusters with radius 30..120 nm. These are treated as single-domain nanoparticles. Magnetic hysteresis, field cooled, zero field cooled and thermoremanent magnetization measurements were performed using an RF SQUID magnetometer and ferromagnetic behaviour was observed. Analysis suggests that the ferromagnetic ordering is associated with defects in a thin surface layer, whose thickness is independent of particle size. Critical radius for single-domain particles, critical radius for coherent rotation, magnetic layer thickness, distance between elementary magnetic moments, saturation magnetization, exchange stiffness constant and anisotropy energy density are also presented.

Ultrasuodatuskalvon modifiointi paineen, lämpötilan ja emäksen avulla

Työssä tutkittiin polymeerisen ultrasuodatuskalvon modifiointimahdollisuuksia prosessiolosuhteita muuttamalla. Kalvon modifioimisella pyritään sen suodatusominaisuuksien muuttumiseen, joka voi lisätä kalvon käyttökohteita ja parantaa kalvon soveltuvuutta tiettyjen yhdisteiden suodatukseen. Hydrofiilisiä, tiukkoja polymeerisiä ultrasuodatuskalvoja on kaupallisesti saatavilla vähän, joten työssä tutkittiin niiden valmistusta modifioimalla markkinoilla olevaa, löysempää, hydrofiilistä, polymeeristä ultrasuodatuskalvoa. Ultrasuodatuskalvo modifioitiin paineen, lämpötilan ja emäksen avulla. Modifioinnin aiheuttamat muutokset voidaan jakaa pysyviin, osittain palautuviin tai palautuviin muutoksiin. Kalvon rakenteen muuttuessa pysyvästi voidaan kalvo modifioida ennen suodatuksen aloittamista. Tällöin modifioinnissa käytetyt olosuhteet eivät vaikuta suodatukseen kuten muissa tapauksissa. Modifioinnin vaikutusta kalvoon voidaan analysoida eri menetelmillä. Näitä ovat esimerkiksi elektronimikroskopia ja kalvon vuon tai retention analysointi. Mikroskooppikuvia ei voida ottaa suodatuksen aikana, vaan kalvosta saada tietoa ainoastaan alku- ja lopputilanteissa suodatusolosuhteista poistettuna. Vuon ja retention avulla saadaan reaaliaikaista tietoa modifioidun kalvon suodatuskapasiteetin ja erotuskyvyn muutoksista. Työssä modifioinnin vaikutusta seurattiin vuo- ja retentiomittausten avulla ja kalvon rakenteessa tapahtuvia muutoksia tutkittiin pyyhkäisyelektronimikroskooppikuvien ja mikrometrimittausten avulla. Korkeampaa painetta tai lämpötilaa käytettäessä havaittiin vuon alenevan modifioitaessa enemmän kuin matalammissa paineissa tai lämpötiloissa. Korkeampi puristuslämpötila kasvatti myös retentiota. Modifiointiolosuhteiden ollessa emäksisiä aleni permeabiliteetti neutraaleissa olosuhteissa tehtyä puristusta enemmän. Myös retentio aleni emäksen avulla tehdyssä modifioinnissa. Kalvon rakenteessa tapahtuneiden muutosten palautuminen riippui modifiointilämpötilasta, korkeassa lämpötilassa modifioidussa kalvossa palautumista ei tapahtunut. Modifioinnin aiheuttamat kalvojen paksuuden muutokset tukivat retentio- ja vuomittauksia. Pyyhkäisyelektronimikroskooppikuvista voitiin havaita kalvon huokosrakenteen puristuneen modifioinnin aikana.

Ring Dark Solitons in Toroidal Bose-Einstein Condensates

In this Thesis, we study various aspects of ring dark solitons (RDSs) in quasi-two-dimensional toroidally trapped Bose-Einstein condensates, focussing on atomic realisations thereof. Unlike the well-known planar dark solitons, exact analytic expressions for RDSs are not known. We address this problem by presenting exact localized soliton-like solutions to the radial Gross-Pitaevskii equation. To date, RDSs have not been experimentally observed in cold atomic gases, either. To this end, we propose two protocols for their creation in experiments. It is also currently well known that in dimensions higher than one, (ring) dark solitons are susceptible, in general, to an irreversible decay into vortex-antivortex pairs through the snake instability. We show that the snake instability is caused by an unbalanced quantum pressure across the soliton's notch, linking the instability to the Bogoliubov-de Gennes spectrum. In particular, if the angular symmetry is maintained (or the toroidal trapping is restrictive enough), we show that the RDS is stable (long-lived with a lifetime of order seconds) in two dimensions. Furthermore, when the decay does take place, we show that the snake instability can in fact be reversible, and predict a previously unknown revival phenomenon for the original (many-)RDS system: the soliton structure is recovered and all the point-phase singularities (i.e. vortices) disappear. Eventually, however, the decay leads to an example of quantum turbulence; a quantum example of the laminar-to-turbulent type of transition.

Maximizing of water recovery in nanofiltration of mine process waters by using antiscalants

Reverse osmosis and nanofiltration are among the most effective and widely used desalination and water softening technologies. They can also be used to treat mining wastewaters and are capable of producing water of extremely high purity, regardless of the high concentrations of toxic heavy metals and extreme pH and salinity. However, challenges with recovering the salts and metals from mining wastewaters in exploitable form, as well as problems with scaling still limit the process efficiency and the ratio of purified water recoverable from process waters. To address the problem of membrane scaling caused by calcium sulfate, batch filtration experiments with the Desal-5 DL nanofiltration membrane, three commercial antiscalants and actual mine process water from a copper mine were performed. The aim of these experiments was to find process conditions where maximum water recovery would be achieved before significant scaling or irreversible membrane fouling would occur and to further improve water recovery by addition of antiscalants. Water recovery of 70 % was reached with the experimental setups by optimizing process conditions. PC-504T antiscaling agent was determined to be the most effective of the three antiscalants used and the addition of 5 ppm of PC-504T allowed the water recovery to be further increased from 70 % to 85 % before major scaling was observed. In these conditions 92 % calcium rejection was achieved.

Imaging Neuroinflammation in Progressive Multiple Sclerosis

Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system CNS), where inflammation and neurodegeneration lead to irreversible neuronal damage. In MS, a dysfunctional immune system causes auto‐reactive lymphocytes to migrate into CNS where they initiate an inflammatory cascade leading to focal demyelination, axonal degeneration and neuronal loss. One of the hallmarks of neuronal injury and neuroinflammation is the activation of microglia. Activated microglia are found not only in the focal inflammatory lesions, but also diffusely in the normal‐appearing white matter (NAWM), especially in progressive MS. The purine base, adenosine is a ubiquitous neuromodulator in the CNS and also participates in the regulation of inflammation. The effect of adenosine mediated via adenosine A2A receptors has been linked to microglial activation, whereas modulating A2A receptors may exert neuroprotective effects. In the majority of patients, MS presents with a relapsing disease course, later advancing to a progressive phase characterised by a worsening, irreversible disability. Disease modifying treatments can reduce the severity and progression in relapsing MS, but no efficient treatment exists for progressive MS. The aim of this research was to investigate the prevalence of adenosine A2A receptors and activated microglia in progressive MS by using in vivo positron emission tomography (PET) imaging and [11C]TMSX and [11C](R)‐PK11195 radioligands. Magnetic resonance imaging (MRI) with diffusion tensor imaging (DTI) was performed to evaluate structural brain damage. Non‐invasive input function methods were also developed for the analyses of [11C]TMSX PET data. Finally, histopathological correlates of [11C](R)‐PK11195 radioligand binding related to chronic MS lesions were investigated in post‐mortem samples of progressive MS brain using autoradiography and immunohistochemistry. [11C]TMSX binding to A2A receptors was increased in NAWM of secondary progressive MS (SPMS) patients when compared to healthy controls, and this correlated to more severe atrophy in MRI and white matter disintegration (reduced fractional anisotropy, FA) in DTI. The non‐invasive input function methods appeared as feasible options for brain [11C]TMSX images obviating arterial blood sampling. [11C](R)‐PK11195 uptake was increased in the NAWM of SPMS patients when compared to patients with relapsing MS and healthy controls. Higher [11C](R)‐PK11195 binding in NAWM and total perilesional area of T1 hypointense lesions was associated with more severe clinical disability, increased brain atrophy, higher lesion load and reduced FA in NAWM in the MS patients. In autoradiography, increased perilesional [11C](R)‐PK11195 uptake was associated with increased microglial activation identified using immunohistochemistry. In conclusion, brain [11C]TMSX PET imaging holds promise in the evaluation of diffuse neuroinflammation in progressive MS. Being a marker of microglial activation, [11C](R)‐ PK11195 PET imaging could possibly be used as a surrogate biomarker in the evaluation of the neuroinflammatory burden and clinical disease severity in progressive MS.

Threat modeling a factory environment using Microsoft Security Development Lifecycle methodology

The number of security violations is increasing and a security breach could have irreversible impacts to business. There are several ways to improve organization security, but some of them may be difficult to comprehend. This thesis demystifies threat modeling as part of secure system development. Threat modeling enables developers to reveal previously undetected security issues from computer systems. It offers a structured approach for organizations to find and address threats against vulnerabilities. When implemented correctly threat modeling will reduce the amount of defects and malicious attempts against the target environment. In this thesis Microsoft Security Development Lifecycle (SDL) is introduced as an effective methodology for reducing defects in the target system. SDL is traditionally meant to be used in software development, principles can be however partially adapted to IT-infrastructure development. Microsoft threat modeling methodology is an important part of SDL and it is utilized in this thesis to find threats from the Acme Corporation’s factory environment. Acme Corporation is used as a pseudonym for a company providing high-technology consumer electronics. Target for threat modeling is the IT-infrastructure of factory’s manufacturing execution system. Microsoft threat modeling methodology utilizes STRIDE –mnemonic and data flow diagrams to find threats. Threat modeling in this thesis returned results that were important for the organization. Acme Corporation now has more comprehensive understanding concerning IT-infrastructure of the manufacturing execution system. On top of vulnerability related results threat modeling provided coherent views of the target system. Subject matter experts from different areas can now agree upon functions and dependencies of the target system. Threat modeling was recognized as a useful activity for improving security.

Clinical and genetic markers in disease progression – a study among subjects with obstructive lung diseases

Asthma, COPD, and asthma and COPD overlap syndrome (ACOS) are chronic pulmonary diseases with an obstructive component. In COPD, the obstruction is irreversible and the disease is progressive. The aim of the study was to define and analyze factors that affected disease progression and patients’ well-being, prognosis and mortality in Chronic Airway Disease (CAD) cohort. The main focus was on COPD and ACOS patients. Retrospective data from medical records was combined with genetic and prospective follow-up data. Smoking is the biggest risk factor for COPD and even after the diagnosis of the disease, smoking plays an important role in disease development and patient’s prognosis. Sixty percent of the COPD patients had succeeded in smoking cessation. Patients who had managed to quit smoking had lower mortality rates and less psychiatric diseases and alcohol abuse although they were older and had more cardiovascular diseases than patients who continued smoking. Genetic polymorphism rs1051730 in the nicotinic acethylcholine receptor gene (CHRNA3/5) associated with heavy smoking, cancer prevalence and mortality in two Finnish independent cohorts consisting of COPD patients and male smokers. Challenges in smoking cessation and higher mortality rates may be partly due to individual patient’s genetic composition. Approximately 50% of COPD patients are physically inactive and the proportion was higher among current smokers. Physically active and inactive patients didn’t differ from each other in regard to age, gender or comorbidities. Bronchial obstruction explained inactivity only in severe disease. Subjective sensation of dyspnea, however, had very strong association to inactivity and was also associated to low health related quality of life (HRQoL). ACOS patients had a significantly lower HRQoL than either the patients with asthma or with COPD even though they were younger than COPD patients, had better lung functions and smaller tobacco exposure.