Platform Architectures for Policy-Based Network Access Control

Tämä diplomityö käsittelee sääntöpohjaisen verkkoon pääsyn hallinnan (NAC) ratkaisuja arkkitehtonisesta näkökulmasta. Työssä käydään läpi Trusted Computing Groupin, Microsoft Corporationin, Juniper Networksin sekä Cisco Systemsin NAC-ratkaisuja. NAC koostuu joukosta uusia sekä jo olemassa olevia teknologioita, jotka auttavat ennalta määriteltyyn sääntökantaan perustuen hallitsemaan suojattuun verkkoon pyrkivien laitteiden tietoliikenneyhteyksiä. Käyttäjän tunnistamisen lisäksi NAC pystyy rajoittamaan verkkoon pääsyä laitekohtaisten ominaisuuksien perusteella, esimerkiksi virustunnisteisiin ja käyttöjärjestelmäpäivityksiin liittyen ja paikkaamaan tietyin rajoituksin näissä esiintyviä puutteita verkkoon pääsyn sallimiseksi. NAC on verraten uusi käsite, jolta puuttuu tarkka määritelmä. Tästä johtuen nykymarkkinoilla myydään ominaisuuksiltaan puutteellisia tuotteita NAC-nimikkeellä. Standardointi eri valmistajien NAC-komponenttien yhteentoimivuuden takaamiseksi on meneillään, minkä perusteella ratkaisut voidaan jakaa joko avoimia standardeja tai valmistajakohtaisia standardeja noudattaviksi. Esitellyt NAC-ratkaisut noudattavat standardeja joko rajoitetusti tai eivät lainkaan. Mikään läpikäydyistä ratkaisuista ei ole täydellinen NAC, mutta Juniper Networksin ratkaisu nousee niistä potentiaalisimmaksi jatkokehityksen ja -tutkimuksen kohteeksi TietoEnator Processing & Networks Oy:lle. Eräs keskeinen ongelma NAC-konseptissa on työaseman tietoverkolle toimittama mahdollisesti valheellinen tietoturvatarkistuksen tulos, minkä perusteella pääsyä osittain hallitaan. Muun muassa tähän ongelmaan ratkaisuna voisi olla jo nykytietokoneista löytyvä TPM-siru, mikä takaa tiedon oikeellisuuden ja koskemattomuuden.

Factors Regulating Chondrogenic Differentiation

Chondrogenesis is a co-ordinated differentiation process in which mesenchymal cells condensate, differentiate into chondrocytes and begin to secrete molecules that form the extracellular matrix. It is regulated in a spatio-temporal manner by cellular interactions and growth and differentiation factors that modulate cellular signalling pathways and transcription of specific genes. Moreover, post-transcriptional regulation by microRNAs (miRNAs) has appeared to play a central role in diverse biological processes, but their role in skeletal development is not fully understood. Mesenchymal stromal cells (MSCs) are multipotent cells present in a variety of adult tissues, including bone marrow and adipose tissue. They can be isolated, expanded and, under defined conditions, induced to differentiate into multiple cell lineages including chondrocytes, osteoblasts and adipocytes in vitro and in vivo. Owing to their intrinsic capability to self-renew and differentiate into functional cell types, MSCs provide a promising source for cell-based therapeutic strategies for various degenerative diseases, such as osteoarthritis (OA). Due to the potential therapeutic applications, it is of importance to better understand the MSC biology and the regulatory mechanisms of their differentiation. In this study, an in vitro assay for chondrogenic differentiation of mouse MSCs (mMSCs) was developed for the screening of various factors for their chondrogenic potential. Conditions were optimized for pellet cultures by inducing mMSC with different bone morphogenetic proteins (BMPs) that were selected based on their known chondrogenic relevance. Characterization of the surface epitope profile, differentiation capacity and molecular signature of mMSCs illustrated the importance of cell population composition and the interaction between different populations in the cell fate determination and differentiation of MSCs. Regulation of Wnt signalling activity by Wnt antagonist sFRP-1 was elucidated as a potential modulator of lineage commitment. Delta-like 1 (dlk1), a factor regulating adipogenesis and osteogenesis, was shown to exhibit stage-specific expression during embryonic chondrogenesis and identified as a novel regulator of chondrogenesis, possibly through mediating the effect of TGF-beta1. Moreover, miRNA profiling demonstrated that MSCs differentiating into a certain lineage exhibit a specific miRNA expression profile. The complex regulatory network between miRNAs and transcription factors is suggested to play a crucial role in fine-tuning the differentiation of MSCs. These results demonstrate that commitment of mesenchymal stromal cells and further differentiation into specific lineages is regulated by interactions between MSCs, various growth and transcription factors, and miRNA-mediated translational repression of lineage-specific genes.

Målarpoeten Ellen Thesleff : teknik och konstnärligt uttryck

The aim of this thesis has been to illustrate the multifaceted talent in Ellen Thesleff's (1869 - 1954) work with particular emphasis on her technique and artistic expression. Why did Ellen Thesleff work withso many techniques? How did the technique affect the expression and what characterizes it? It would also be of interest to gather some idea of Ellen Thesleff's position among other artists. The investigation covers a representative selection of about 60 pictures, using nine different techniques, primarily as oils, woodcuts and monumental painting. The pictures illustrate three periods of time, the natural (1890 - 1905), the colourful (1906 - 1927) and the free period (1928 -1950). I describe the pictures in regard to their conception and subject matter and scrutinize their formal creation. Thereafter, I investigate the painting technique and artistic expression of each picture and position it, where applicable, in relation to other art. Ellen Thesleff's artistic quality is discussed in relation to her techniques and expression. Thesleff consciously chooses different techniques for related subjects in order to vary the expression. The progressing evolution within individual techniques and a cross-fertilization between them has evidently contributed to raising her artistic quality. I have studied how the techniques influence expression and found it possible to identify certain characteristic styles during the three periods: first a natural painting technique which reminds one of both French realism, paintings reflecting the Nordic mood and atmosphere, and symbolism, ascetism and synthetism; later an expressive Thesleff colourism with brilliant over- and underpainting in contrasting colours and last a free decorative painting in lines, with symbolistic undertones. Most characteristic is the lyrical expression which seems to be a common theme throughout Thesleff's entire artistry. I have found that Ellen Thesleff in her works had herown personal style compared to her contemporaries. Despite deep knowledge of styles and techniques she continually creates art from her inner self and with her own personal brush signature.

Identification and characterization of CIP2A as a novel oncogenic inhibitor of PP2A

Inhibition of the tumor suppressor protein phosphatase 2A (PP2A) activity has been identified as one of the five key alterations required for human cell transformation. Regardless of this crucial role in human cancer development, the detailed mechanisms by which PP2A inhibition occurs in human cancers remain largely uncharacterized. PP2A regulates a plethora of cellular signaling cascades. One of the targets of PP2A is Myc oncoprotein, which is destabilized and degraded in response to PP2A-mediated dephosphorylation of Myc serine 62. In this study we identify Cancerous Inhibitor of PP2A (CIP2A) as a previously uncharacterized endogenous inhibitor of PP2A in human cancer cells. CIP2A inhibits PP2A activity leading to subsequent stabilization of the Myc protein. CIP2A promotes malignant growth of cancer cells in vitro and xenograft tumor formation in vivo and is overexpressed in cancer. Moreover, we explored the effect of CIP2A on global transcriptional profiles and validated a CIP2A-dependent transcriptional signature. Analysis of the CIP2A signature revealed both Myc-dependent and -independent functions for CIP2A. Importantly, we demonstrate that the CIP2A signature has clinical relevance in human breast cancer subtypes. Finally, we identify the genes potentially mediating the long-term growth suppression in CIP2A depleted cancer cells. Taken together, this work identifies CIP2A as a novel human oncoprotein and describes its function in cancer cells. These results may open novel possibilities for patient stratification and therapeutic intervention of cancer.

Reactive oxygen species in inflammation

Chronic inflammation is the underlying cause of many common disabling conditions such as rheumatoid arthritis (RA), multiple sclerosis, coeliac disease, type I diabetes and coronary artery disease. NOX2 complex derived reactive oxygen species (ROS) are known to regulate joint inflammation in rats and mice, and additionally recent genetic evidence associates phagocyte ROS and the development RA in humans. Ncf1mutated mice have lost the functionality of their NOX2 complex and thus have no phagocyte ROS production. These mice suffer from exacerbated arthritis. The immune suppressive effect of the NOX2 complex derived ROS is mediated by monocytes/macrophages that downregulate the activation of autoreactive T cells. The aim of this thesis was to study how ROS modulate immune responses in different arthritis models and in tumor development. Additionally, genome wide gene expression profiling was carried out to assess the global effects of NOX2 complex derived ROS. Firstly, these results confirmed the potent anti-inflammatory nature of phagocyte ROS in arthritis models that were driven by the adaptive immune system. Secondly, arthritis models with predominantly innate immunity induced pathophysiology were moderately enhanced by phagocyte, more specifically, neutrophil derived ROS. Thirdly, the ROS induced immune suppression mediated by the adaptive immune system allowed development of bigger implanted tumors, while phagocyte ROS production did not affect the development of spontaneously growing tumors. Lastly, genome wide gene expression analysis revealed that both humans and mice with abrogated phagocyte NOX2 complex ROS production had an enhanced type I interferon signature in blood, reflecting their hyperinflammatory immune status.

Organisaatioprosessin muutos

Tutkimuksessa tarkasteltiin sähköiseen arkistointiin siirtymisen muutosprosessia Lahden kaupungin organisaatiossa vuosien 2013 – 2015 välillä. Tavoitteena oli sel-vittää kunnallisen organisaation sähköiseen arkistointiin siirtymisen vaatiman muu-tosprosessin toteutuksen ratkaisut sekä muutoksen vaikutukset hallinnollisiin pro-sesseihin. Työ on metodologialtaan kvalitatiivinen tapaustutkimus, jonka tutkimus-aineistona käytettiin haastatteluja sekä organisaation sisäisiä asiakirjoja. Tutkimuk-sessa haastateltiin kohdeorganisaation muutoksen suunnittelun sekä toteutuksen avainhenkilöitä. Tutkimustulokset osoittivat, että muutosprosessi noudatti tieteellisen tutkimuksen kuvaamaa muutoksen vaiheittaista etenemistä, vaikkakin vaiheet me-nivät osittain päällekkäin. Muutosprosessissa tulisi tulosten perusteella kiinnittää huomiota muutoksessa ilmenevien vastakkaisten mielipiteiden vaikutukseen muu-toksen hidastavana tekijänä ja ottaa jo suunnitteluvaiheessa tämä huomioon esi-merkiksi miettimällä toimiva kannustinjärjestelmä tukemaan muutoksen toteutusta. Merkittävimmät havainnot prosessien muuttumiseen liittyen olivat, että arkistointia ei voi ajatella erillisenä prosessina asiakirjan elinkaaren lopussa, vaan sähköisessä toimintaympäristössä arkistointi sulautuu kiinteäksi osaksi organisaation kaikkea toimintaa. Arkistoinnin tapaa muutettaessa on otettava huomioon eri toiminnoille asetetut vaatimukset, kuten tietoturva, allekirjoitus, arkistointi, arkistossa asiointi se-kä hävitys. Sähköiseen arkistointiin siirtyminen muuttaa organisaation eri toimijoi-den rooleja ja jossain kohden tuo säästöjä, mutta toisaalta vaatii organisaatiolta muuttumiskykyä. Osaltaan muutos vauhdittaa myös erilaisten toimintojen keskittä-misen mahdollisuuksia.

Targeting oncogenic signaling proteins : new insights using a FRET-based chemical biology approach

Cancer affects more than 20 million people each year and this rate is increasing globally. The Ras/MAPK-pathway is one of the best-studied cancer signaling pathways. Ras proteins are mutated in almost 20% of all human cancers and despite numerous efforts, no effective therapy that specifically targets Ras is available to date. It is now well established that Ras proteins laterally segregate on the plasma membrane into transient nanoscale signaling complexes called nanoclusters. These Ras nanoclusters are essential for the high-fidelity signal transmission. Disruption of nanoclustering leads to reduction in Ras activity and signaling, therefore targeting nanoclusters opens up important new therapeutic possibilities in cancer. This work describes three different studies exploring the idea of membrane protein nanoclusters as novel anti-cancer drug targets. It is focused on the design and implementation of a simple, cell-based Förster Resonance Energy Transfer (FRET)-biosensor screening platform to identify compounds that affect Ras membrane organization and nanoclustering. Chemical libraries from different sources were tested and a number of potential hit molecules were validated on full-length oncogenic proteins using a combination of imaging, biochemical and transformation assays. In the first study, a small chemical library was screened using H-ras derived FRET-biosensors. Surprisingly from this screen, commonly used protein synthesis inhibitors (PSIs) were found to specifically increase H-ras nanoclustering and downstream signalling in a H-ras dependent manner. Using a representative PSI, increase in H-ras activity was shown to induce cancer stem cell (CSC)-enriched mammosphere formation and tumor growth of breast cancer cells. Moreover, PSIs do not increase K-ras nanoclustering, making this screening approach suitable for identifying Ras isoform-specific inhibitors. In the second study, a nanoncluster-directed screen using both H- and K-ras derived FRET biosensors identified CSC inhibitor salinomycin to specifically inhibit K-ras nanocluster organization and downstream signaling. A K-ras nanoclusteringassociated gene signature was established that predicts the drug sensitivity of cancer cells to CSC inhibitors. Interestingly, almost 8% of patient tumor samples in the The Cancer Genome Atlas (TCGA) database had the above gene signature and were associated with a significantly higher mortality. From this mechanistic insight, an additional microbial metabolite screen on H- and K-ras biosensors identified ophiobolin A and conglobatin A to specifically affect K-ras nanoclustering and to act as potential breast CSC inhibitors. In the third study, the Ras FRET-biosensor principle was used to investigate membrane anchorage and nanoclustering of myristoylated proteins such as heterotrimeric G-proteins, Yes- and Src-kinases. Furthermore, Yes-biosensor was validated to be a suitable platform for performing chemical and genetic screens to identify myristoylation inhibitors. The results of this thesis demonstrate the potential of the Ras-derived FRETbiosensor platform to differentiate and identify Ras-isoform specfic inhibitors. The results also highlight that most of the inhibitors identified predominantly perturb Ras subcellular distribution and membrane organization through some novel and yet unknown mechanisms. The results give new insights into the role of Ras nanoclusters as promising new molecular targets in cancer and in stem cells.

CLEVER-1 as an immune suppressive molecule

The immune response and immune suppression are equally essential for the immune system to protect the host against an infection and to protect self-molecules in different pathophysiological conditions. Pregnancy is one of the conditions where the maternal immune system remains resistant against microbes and yet attains tolerance to protect the fetus, whose genetic material differs partially from the mother’s. However, if the balance of immune suppression is not precise in the host it can favor conditions which lead to diseases, such as cancer and autoimmune disorders. This study was initiated to investigate the expression and functions of CLEVER-1/Stabilin-1, a multifunctional protein expressed on subsets of endothelial cells and type II macrophages, as an immune suppressive molecule. Firstly, the expression of CLEVER-1/stabilin-1 and its function in human placental macrophages were examined. Secondly, the expression profile and functional significance of stabilin-1 on healthy human monocytes was investigated. The results clarified the expression of CLEVER-1/stabilin-1 on placental macrophages, and verified that CLEVER-1/stabilin-1 functions as an adhesion and scavenging molecule on these cells. The data from normal monocytes revealed that the monocytes with low stabilin-1 expression carried a pro-inflammatory gene signature, and that stabilin-1 can directly or indirectly regulate pro-inflammatory genes in monocytes. Finally, it was shown that monocyte CLEVER-1/stabilin-1 dampens IFN production by T cells. To conclude, CLEVER-1/stabilin-1 is defined as an immune suppressive molecule on monocytes and macrophages. Strikingly, anti-stabilin-1 antibodies may have the potential to promote the Th1 dependent inflammatory response and counteract the tumor induced immune suppression.

Analogisen tunnisteanalyysin käyttö korjauspalveluissa mikropiirien ylijännitteestä aiheutuneiden vaurioiden paikantamisen

Korjauspalveluissa aikaa vieviä tapauksia ovat mikropiirien vaikeasti paikannettavat viat. Tällaista vianetsintää varten yrityksemme oli ostanut Polar Fault Locator 780 –mittalaitteen, jolla voidaan mitata mikropiirien toimintaa käyttämällä analogista tunnisteanalyysiä. Diplomityön tavoitteena oli selvittää, miten mittaustapaa voidaan käyttää korjauspalveluissa. Tutkintaa lähestyttiin joidenkin tyypillisten komponenttien näkökulmasta, mutta pääpaino oli mikropiireissä. Joitain mikropiirejä vaurioitettiin tahallisesti, jolloin mittaustulokset uusittiin ja tutkittiin miten vaurioituminen näkyy mittaustuloksissa. Tutkimusmenetelmänä oli kirjallisuus ja empiirinen kokeellisuus. Diplomityön tuloksena oli, että tätä mittaustapaa käyttämällä mikropiirien kuntoa voidaan tutkia. Ongelmiksi osoittautuivat alkuperäinen oletus mittalaitteen tuloksien tulkinnasta ja taustamateriaalin heikko saatavuus. Täten mittalaite parhaiten soveltuu tilanteisiin, joissa sen antamia tuloksia verrataan suoraan toisen toimivaksi tunnetun yksikön mittaustuloksiin. Vaurioitettaessa komponenteissa oli kuitenkin havaittavissa selvä poikkeavuus.