Characterizing cancer cell signaling at the protein level : from targeted to proteome and phosphoproteome­‐wide analyses

Proteins are the effectors of cellular functions and the constituting elements of cellular signaling cascades. The ability to analyze the abundances and the dynamics of proteins is central to dissect cellular signaling and its effects on cell physiology. The aim of this thesis is to gain insight into protein level regulatory mechanisms that contribute to the development of cancer, by optimizing and employing targeted and largescale methods. Specifically, to examine mechanisms regulating protein stability, localization, protein-protein interactions, and to characterize targets of a protein phosphatase enzymatic activity. Additionally, to optimize a workflow for quantitative phosphoproteomics analysis with the goal of improving the sensitivity and lower the requirement in terms of sample quantity of current methods. Study I elucidated a mechanism by which S100A4 interacts with p53 in the nucleus thereby promoting its degradation, and the effects of this interaction on the growth and survival of lung and cervix adenocarcinoma cell lines, by employing targeted methods for the analyses of protein stability, protein localization and protein-protein interactions. Study II elucidated a mechanism by which TRAP promotes metastasis-related cell properties in breast cancer cells via the TGFβ-pathway and CD44, by a combination of proteomics and phosphoproteomics analyses with targeted methods. Furthermore, a moderate-depth phosphoproteomic profiling of TRAP overexpressing cells was achieved by peptide fractionation by high-resolution isoelectric focusing (HiRIEF) on IPG strips pH range 2.5- 3.7, and provided a list of putative targets of TRAP phosphatase activity. Study III developed a workflow for in-depth quantitative phosphoproteomics analysis based on high-resolution isoelectric focusing (HiRIEF) fractionation on a wide pH range (2.5-10). The workflow employs phospho-enrichment by titanium dioxide coupled with isobaric labeling by TMT, and provides for good analytical depth and sensitivity, requiring a low amount of starting material. Application of this workflow for the analysis of cervix adenocarcinoma cells HeLa revealed 1,264 novel phosphorylation sites, of which 165 phospho-sites that are suggested to have a regulatory function during the mitotic phase, based on kinase-association analysis. In summary, the work presented in this thesis contributes to the collective effort of improving and applying targeted and large-scale methods for the analysis of protein level regulatory mechanisms, particularly by focusing on the optimization of a workflow for phosphoproteomics analysis. Development of these methods and improvements in integrating discovery and validation efforts, will be central in the coming years and offer unprecedented opportunities for increasing our understanding of life and to discover new treatments and cures for diseases

Fee Arrangements and Fee Shifting: Lessons From the Experience in Ontario

About one-third of oestrogen receptor alpha-positive breast cancer patients treated with tamoxifen relapse. Here we identify the nuclear receptor retinoic acid receptor alpha as a marker of tamoxifen resistance. Using quantitative mass spectrometry-based proteomics, we show that retinoic acid receptor alpha protein networks and levels differ in a tamoxifen-sensitive (MCF7) and a tamoxifen-resistant (LCC2) cell line. High intratumoural retinoic acid receptor alpha protein levels also correlate with reduced relapse-free survival in oestrogen receptor alpha-positive breast cancer patients treated with adjuvant tamoxifen solely. A similar retinoic acid receptor alpha expression pattern is seen in a comparable independent patient cohort. An oestrogen receptor alpha and retinoic acid receptor alpha ligand screening reveals that tamoxifen-resistant LCC2 cells have increased sensitivity to retinoic acid receptor alpha ligands and are less sensitive to oestrogen receptor alpha ligands compared with MCF7 cells. Our data indicate that retinoic acid receptor alpha may be a novel therapeutic target and a predictive factor for oestrogen receptor alpha-positive breast cancer patients treated with adjuvant tamoxifen

Käytösoireisen muistisairaan lääkkeettömät hoitotyön keinot : Opas sairaanhoitajille

Aiheena opinnäytetyölle oli Käytösoireisen muistisairaan lääkkeettömät hoitotyön keinot – opas sairaanhoitajille. Opinnäytetyön tavoitteena oli tuottaa tietoa muistisairaan käytöshäiriöiden syistä ja lisäksi lisätä ymmärrystä sairaanhoitajille muistisairaan haasteellisen käytöksen lisääntymisestä. Opinnäytetyön tarkoituksena oli tuottaa yhteistyökumppanille opas, joka sisältää yleisimmät muistisairaudet ja niihin liittyvien käytöshäiriöiden ilmenemismuodot aiheuttajineen ja vaikuttavine tekijöineen. Tämän lisäksi opas sisältää hoitotyön keinoja, joiden avulla sairaanhoitaja pystyy havainnoimaan ja lieventämään muistisairaan käytösoireita. Opinnäytetyö on tehty yhteistyössä Suupohjan peruspalveluliikelaitoskuntayhtymän hoidon ja hoivan alueen kanssa. Etenevien muistisairauksien diagnostiikkaan kuuluu käytösoireiden lisääntyminen. Käytösoireita on muistisairauksien kaikissa vaiheissa ja niiden ilmaantuminen voi johtaa liialliseen ja turhaan rauhoittavien lääkkeiden määräämiseen ja käyttöön, on kuitenkin hyvä huomioida kokonaisvaltainen hoito, joka on potilaan yksilöllisen tilanteen huomioon ottava. Muistisairaiden määrä kasvaa nopeasti, varhaisen diagnosoinnin avulla pystytään ylläpitämään sairastuneen toimintakykyä ja huomioimaan sairastuneen oman elämänlaadun pysyminen hyvänä, unohtamatta hänen läheisiään. Käytösoireisen potilaan hoitolinja tulisi valita arvioimalla oireita ja selvittämällä niiden syy. Lääkkeettömän hoidon tarkoitus on, että muistisairaasta huolehditaan kokonaisvaltaisesti ja mahdollisimman hyvin hänen tarpeensa huomioon ottaen. Sairastuneen toimintakyvyn tukeminen on tärkeää, silloin hän tuntee olonsa turvatuksi ja arvostetuksi. Hyvien elämäntapojen huomioiminen, riittävän unen ja aktiviteetin turvaaminen tukevat sairastuneen tasapainon tunnetta. Käytöshäiriöiden syntyyn vaikuttaa myös ympäristössä tapahtuvat muutokset. Sairaanhoitajan on tärkeä luoda sairastuneelle tässä tilanteessa rauhallinen ja turvattu ympäristö.The subject for the thesis is non-drug nursing methods of a patient with memory disease and behavioural disorder. The aim of the thesis was to provide information on the causes of behavioural disorders of a patient with memory disease, and in addition, to increase nurses’ understanding about the negative behaviour of memory patients. The purpose of the thesis was to produce a guide containing the most common memory disorders and related manifestations of behavioural disorders, with their causes and contributing factors. In addition, the guide includes nursing tools that help the nurse to observe and mitigate the behavioural disorders of a patient with a memory disease. The thesis has been carried out in cooperation with the treatment and care area of The Suupohja Area Health and Social Services Joint Municipal Board. The diagnostics for progressive memory diseases include an increase in behavioural disorders. There are behavioural disorders at all stages of memory disorders, and their appearance may lead to the prescription and use of excessive and unnecessary medication. However, it is good to take into account the holistic treatment that is appropriate to the patient's individual condition. The number of patients with memory disease is increasing rapidly. With early diagnosis, it is pos-sible to maintain patients’ functional ability and take account of their quality of life, not forgetting their close relatives. The treatment line for the patient with behavioural disorder should be selected by evaluating the symptoms and finding out their cause. The purpose of non-drug nursing is that the patient with memory disorder is taken care of comprehensively, and his or her needs are taken into account as well as possible. Supporting the functional ability of the patient is important, and he or she feels secure and appreciated. Paying attention to a good lifestyle, ensuring adequate sleep and activity support the balance feeling of the patient. Changes in the environment also affect the appearance of behavioural disorders. It is important for the nurse to create a calm and secure environment for the patient in this situation

Efficacy of a Cancer Vaccine against ALK-Rearranged Lung Tumors

Non-small cell lung cancer (NSCLC) harboring chromosomal rearrangements of the anaplastic lymphoma kinase (ALK) gene is treated with ALK tyrosine kinase inhibitors (TKIs), but is successful for only a limited amount of time; most cases relapse due to the development of drug resistance. Here we show that a vaccine against ALK induced a strong and specific immune response that both prophylactically and therapeutically impaired the growth of ALK-positive lung tumors in mouse models. The ALK vaccine was efficacious also in combination with ALK TKI treatment and significantly delayed tumor relapses after TKI suspension. We found that lung tumors containing ALK rearrangements induced an immunosuppressive microenvironment, regulating the expression of PD-L1 on the surface of lung tumor cells. High PD-L1 expression reduced ALK vaccine efficacy, which could be restored by administration of anti-PD-1 immunotherapy. Thus, combinations of ALK vaccine with TKIs and immune checkpoint blockade therapies might represent a powerful strategy for the treatment of ALK-driven NSCLC

Initial examples of the SOLUS multimodal potential

We present initial evidence of the SOLUS potential for the multimodal non-invasive diagnosis of breast cancer by describing the correlation between optical and standard radiological data and analyzing a case study

Atti normativi

Raccolta aggiornata delle fonti concernenti la giustizia costituzionale

Giustizia costituzionale, 5. ed.

Manuale aggiornato e di livello specialistico di giustizia costituzional

Giustizia costituzionale

La trattazione è quella di un manuale riferito ad una parte specifica del diritto costituzionale, quale la giustizia costituzionale, ormai da tempo assurta a disciplina autonoma