Quantitative proteomics in the characterization of T helper lymphocyte differentiation

The term proteome is used to define the complete set of proteins expressed in cells or tissues of an organism at a certain timepoint. Respectively, proteomics is used to describe the methods, which are used to study such proteomes. These methods include chromatographic and electrophoretic techniques for protein or peptide fractionation, mass spectrometry for their identification, and use of computational methods to assist the complicated data analysis. A primary aim in this Ph.D. thesis was to set-up, optimize, and develop proteomics methods for analysing proteins extracted from T-helper (Th) lymphocytes. First, high-throughput LC-MS/MS and ICAT labeling methods were set-up and optimized for analysing the microsomal fraction proteins extracted from Th lymphocytes. Later, iTRAQ method was optimized to study cytokine regulated protein expression in the nuclei of Th lymphocytes. High-throughput LC-MS/MS analyses, like ICAT and iTRAQ, produce large quantities of data and robust software and data analysis pipelines are needed. Therefore, different software programs used for analysing such data were evaluated. Moreover, a pre-filtering algorithm was developed to classify good-quality and bad-quality spectra prior to the database searches. Th-lymphocytes can differentiate into Th1 or Th2 cells based on surrounding antigens, co-stimulatory molecules, and cytokines. Both subsets have individual cytokine secretion profiles and specific functions. Th1 cells participate in the cellular immunity against intracellular pathogens, while Th2 cells have important role in the humoral immunity against extracellular parasites. An abnormal response of Th1 and Th2 cells and imbalance between the subsets are charasteristic of several diseases. Th1 specific reactions and cytokines have been detected in autoimmune diseases, while Th2 specific response and cytokine profile is common in allergy and asthma. In this Ph. D. thesis mass spectrometry-based proteomics was used to study the effects of Th1 and Th2 promoting cytokines IL-12 and IL-4 on the proteome of Th lymphocytes. Characterization of microsomal fraction proteome extracted from IL-12 treated lymphobasts and IL-4 stimulated cord blood CD4+ cells resulted in finding of cytokine regulated proteins. Galectin-1 and CD7 were down-regulated in IL-12 treated cells, while IL-4 stimulation decreased the expression of STAT1, MXA, GIMAP1, and GIMAP4. Interestingly, the transcription of both GIMAP genes was up-regulated in Th1 polarized cells and down-regulated in Th2 promoting conditions.

Yksintyöskentely : yksintekeminen työmenetelmänä av-tuotannoissa

Opinnäytetyössäni käsittelen yksintekemiseen perustuvaa työskentelytapaa. Tutkin dokumentin, tv-ohjelmien ja uutisten parissa toimivia yksintekijöitä. Määrittelen yksintyöskentelijäksi sellaisen tekijän, joka kuvauksen lisäksi ohjaa tai haastattelee yksin eli on kuvaustilanteessa yksin. Tämän lisäksi yksintyöskentelijä voi tehdä kaikkia muita audiovisuaalisen tuotannon työtehtäviä kuten tuottaa, leikata ja käsitellä ääntä. Tarkastelen aihetta analysoimalla omaa tekemistäni yksintyöskentelynä tekemässäni dokumentissa sekä haastattelemalla kolmea erilaista yksintekijää: dokumentaristia, ajankohtaisohjelman tekijää ja uutisten tekijää. Kartoitan heidän motivaatiotaan ja toimintatapojaan kaikissa av-tuotannon vaiheissa. Haastattelujen sekä omien kokemusten perusteella voi todeta, että yksintyöskentelijälle kevyt ja helppokäyttöinen mutta samalla laadukas kuvauskalusto on erittäin tärkeä. Yksintyöskentely on edullinen tuotantotapa, joka mahdollistaa aikataulun joustavuuden ja nopean liikkeelle lähdön sekä mahdollisuuden intiimiin kuvaukseen ilman isoa kuvausryhmää ja välineitä. Yksintekemistä puolestaan hankaloittaa moneen asiaan keskittyminen yhtä aikaa sekä joissain tilanteissa välineiden rajoittuneisuus. Yksintyöskentelijä joutuu myös kantamaan vastuun yksin monista eri työvaiheista. Kaikille haastatelluilleni hakeutuminen yksintyöskentelijäksi on ollut luonnollinen tai sopiva vastaan tullut vaihtoehto. Eri alueilta uransa aloittaneet yksintyöskentelijät olivat kaikki motivoituneita ja monipuolisesti ammattitaitoisia. Heille yksintyöskentely oli toimiva tapa tehdä dokumentteja ja tv-ohjelmia.

Randomized study of reduced-intensity chemotherapy combined with imatinib in adults with Ph-positive acute lymphoblastic leukemia.

In this study, we randomly compared high doses of the tyrosine kinase inhibitor imatinib combined with reduced-intensity chemotherapy (arm A) to standard imatinib/hyperCVAD (cyclophosphamide/vincristine/doxorubicin/dexamethasone) therapy (arm B) in 268 adults (median age, 47 years) with Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia (ALL). The primary objective was the major molecular response (MMolR) rate after cycle 2, patients being then eligible for allogeneic stem cell transplantation (SCT) if they had a donor, or autologous SCT if in MMolR and no donor. With fewer induction deaths, the complete remission (CR) rate was higher in arm A than in arm B (98% vs 91%; P = .006), whereas the MMolR rate was similar in both arms (66% vs 64%). With a median follow-up of 4.8 years, 5-year event-free survival and overall survival (OS) rates were estimated at 37.1% and 45.6%, respectively, without difference between the arms. Allogeneic transplantation was associated with a significant benefit in relapse-free survival (hazard ratio [HR], 0.69; P = .036) and OS (HR, 0.64; P = .02), with initial white blood cell count being the only factor significantly interacting with this SCT effect. In patients achieving MMolR, outcome was similar after autologous and allogeneic transplantation. This study validates an induction regimen combining reduced-intensity chemotherapy and imatinib in Ph+ ALL adult patients and suggests that SCT in first CR is still a good option for Ph+ ALL adult patients. This trial was registered at www.clinicaltrials.gov as #NCT00327678.

Molecular mechanism of a green-shifted, pH-dependent red fluorescent protein mKate variant.

Fluorescent proteins that can switch between distinct colors have contributed significantly to modern biomedical imaging technologies and molecular cell biology. Here we report the identification and biochemical analysis of a green-shifted red fluorescent protein variant GmKate, produced by the introduction of two mutations into mKate. Although the mutations decrease the overall brightness of the protein, GmKate is subject to pH-dependent, reversible green-to-red color conversion. At physiological pH, GmKate absorbs blue light (445 nm) and emits green fluorescence (525 nm). At pH above 9.0, GmKate absorbs 598 nm light and emits 646 nm, far-red fluorescence, similar to its sequence homolog mNeptune. Based on optical spectra and crystal structures of GmKate in its green and red states, the reversible color transition is attributed to the different protonation states of the cis-chromophore, an interpretation that was confirmed by quantum chemical calculations. Crystal structures reveal potential hydrogen bond networks around the chromophore that may facilitate the protonation switch, and indicate a molecular basis for the unusual bathochromic shift observed at high pH. This study provides mechanistic insights into the color tuning of mKate variants, which may aid the development of green-to-red color-convertible fluorescent sensors, and suggests GmKate as a prototype of genetically encoded pH sensors for biological studies.

Conversa amb Th. S. Popkewitz: els ensenyants i la reforma de la formació del professorat als EUA, professionalització i poder

La meva hipòtesi és que els supòsits socials i epistemològics que s'han incorporat a les reformes nord-americanes de formació del profes.sorat tendeixen a mistificar la complexitat social i política de I'escolarització i a descontextualitzar les profundes qüestions sobre el poder i el coneixement a I'ensenyament. Les regles, els programes de recerca, els informes de les administracions estan expressats en els termes més humanistes i progressistes possibles. Pretenen portar-nos a tots la democracia, generar un creixement económic i una renovació cultural, i també promoure el desenvolupament individual. Peró a través del recorregut deis somnis i la retórica progressista hom pot observar la creació de tecnologies que s'internalitzen i organitzen la vida quotidiana, Ilurs esperances i anhels. Th .. S. Popkewitz (1993)

Planering av tidtabell, kostnader och arbetsplats för småhus

Det här examensarbete gjordes åt Seasong Travels Oy. I arbetet gjordes en kostnadsplan, tidtabell och dispositionsplan för en stugby på Norrkullalandet. Arbetet börjades med att bekanta sej med byggbranschens facklitteratur och ta reda på vilka faktorer som påverkar byggprojektets slutliga kostnader och tidtabell. På basen av områdets litteratur valdes kostnadsberäkningssättet att utföras med hjälp av Talo-80 nomenklatur och göra en byggnadsdelskalkyl. I arbetets undersökningsskede delades stugorna upp enligt Talo-80 nomenklatur i enskilda byggnadsdelar, varefter byggnadsdelarna prissattes. En tidtabell gjordes när alla byggnadsdelar var uppdelade och med hjälp av områdets litteratur räknades den effektiva byggtiden ut. Dispositionsplanen gjordes genom att studera områdets facklitteratur och själva byggplatsen.

Archives de l'art français : recueil de documents inédits relatifs à l'histoire des arts en France / publié sous la direction. de Ph. de Chennevières

1858 (T5)-1859.

Archives de l'art français : recueil de documents inédits relatifs à l'histoire des arts en France / publié sous la direction. de Ph. de Chennevières

1851 (T1)-1852.

Archives de l'art français : recueil de documents inédits relatifs à l'histoire des arts en France / publié sous la direction. de Ph. de Chennevières

1857 (T5)-1858.

Archives de l'art français : recueil de documents inédits relatifs à l'histoire des arts en France / publié sous la direction. de Ph. de Chennevières

1855 (T4)-1856.