Anti-TNF treatment in juvenile idiopathic arthritis and associated uveitis : Clinical perspectives on growth, uveitis, and drug survival

Juvenile idiopathic arthritis (JIA) is a heterogeneous group of childhood chronic arthritides, associated with chronic uveitis in 20% of cases. For JIA patients responding inadequately to conventional disease-modifying anti-rheumatic drugs (DMARDs), biologic therapies, anti-tumor necrosis factor (anti-TNF) agents are available. In this retrospective multicenter study, 258 JIA-patients refractory to DMARDs and receiving biologic agents during 1999-2007 were included. Prior to initiation of anti-TNFs, growth velocity of 71 patients was delayed in 75% and normal in 25%. Those with delayed growth demonstrated a significant increase in growth velocity after initiation of anti-TNFs. Increase in growth rate was unrelated to pubertal growth spurt. No change was observed in skeletal maturation before and after anti-TNFs. The strongest predictor of change in growth velocity was growth rate prior to anti-TNFs. Change in inflammatory activity remained a significant predictor even after decrease in glucocorticoids was taken into account. In JIA-associated uveitis, impact of two first-line biologic agents, etanercept and infliximab, and second-line or third-line anti-TNF agent, adalimumab, was evaluated. In 108 refractory JIA patients receiving etanercept or infliximab, uveitis occurred in 45 (42%). Uveitis improved in 14 (31%), no change was observed in 14 (31%), and in 17 (38%) uveitis worsened. Uveitis improved more frequently (p=0.047) and frequency of annual uveitis flares was lower (p=0.015) in those on infliximab than in those on etanercept. In 20 patients taking adalimumab, 19 (95%) had previously failed etanercept and/or infliximab. In 7 patients (35%) uveitis improved, in one (5%) worsened, and in 12 (60%) no change occurred. Those with improved uveitis were younger and had shorter disease duration. Serious adverse events (AEs) or side-effects were not observed. Adalimumab was effective also in arthritis. Long-term drug survival (i.e. continuation rate on drug) with etanercept (n=105) vs. infliximab (n=104) was at 24 months 68% vs. 68%, and at 48 months 61% vs. 48% (p=0.194 in log-rank analysis). First-line anti-TNF agent was discontinued either due to inefficacy (etanercept 28% vs. infliximab 20%, p=0.445), AEs (7% vs. 22%, p=0.002), or inactive disease (10% vs. 16%, p=0.068). Females, patients with systemic JIA (sJIA), and those taking infliximab as the first therapy were at higher risk for treatment discontinuation. One-third switched to the second anti-TNF agent, which was discontinued less often than the first. In conclusion, in refractory JIA anti-TNFs induced enhanced growth velocity. Four-year treatment survival was comparable between etanercept and infliximab, and switching from first-line to second-line agent a reasonable therapeutic option. During anti-TNF treatment, one-third with JIA-associated anterior uveitis improved.

Resistance responses blocking systemic movement of Potato virus A in potato

Peruna kestää A-virusta estämällä sen leviämistä Peruna on maissin ohella maailman kolmanneksi tärkein ravintokasvi vehnän ja riisin jälkeen. Perunaa lisätään kasvullisesti mukuloita istuttamalla, jolloin virukset siirtyvät sairaiden siemenmukuloiden välityksellä kasvukaudesta toiseen. Virustauteja voi torjua ainoastaan terveen siemenperunan ja kestävien lajikkeiden avulla. Kestävyys perustuu usein siihen, että kasvi estää viruksen leviämisen tartuntakohdasta välttyäkseen virustaudilta. Tässä työssä tutkittiin kolmea perunan A-viruksen (PVA) liikkumista estävää kestävyysmekanismia perunassa. Lisäksi työn kokeelliseen osaan oleellisesti kuuluvaa virustartutusta varten kehitettiin uusi paranneltu versio geenipyssystä. Tämä itse rakennettu laite optimoitiin PVA:n tartuttamiseen mahdollisimman helposti ja pienin käyttökustannuksin. Tutkimuksen kohteena olleessa perunan risteytysjälkeläistössä oli PVA:ta kestäviä kasveja (ryhmä nnr), jotka estivät viruksen liikkumisen aiheuttamatta oireita tartutuskohdassa, sekä kasveja, joissa PVA aiheutti kuolioläikkinä näkyvän yliherkkyysvasteen (ryhmä HR). Molemmissa kestävyystyypeissä virus pystyi monistumaan ja leviämään solusta soluun paikallisesti, mutta liikkuminen muihin kasvinosiin nilan kautta estyi. Ryhmän nnr kasveissa PVA-tartunta ei aiheuttanut tilastollisesti merkitsevää muutosta useimpien geenien ilmenemiseen tartuntakohdassa. Ainoastaan geeniperhe, joka ilmentää tiettyä proteinaasi-inhibiittoria (PI), reagoi PVA:han 24 tuntia tartutuksesta. Kun tämän PVA:han reagoivan geeniperheen jäsenet hiljennettiin nnr- perunalinjoissa, ne muuttuivat alttiiksi PVA:lle ja virus levisi tartuntakohdasta muihin kasvinosiin. Tulos osoittaa, että PI on viruskestävyystekijä. Lisäksi muut tutkimuksessa saadut tulokset tukevat mahdollisuutta, että PI estää PVA:n P1-proteinaasin toimintaa. HR-linjoissa todettiin erilaisiin puolustusvasteisiin liittyvien PR-geenien aktivoitumista PVA-tartunnan seurauksena, mutta myös ilman sitä kasvien kasvettua mullassa noin neljä viikkoa. Sen sijaan solukkoviljelyssä tai vasta kaksi viikkoa mullassa kasvaneissa kasveissa vastaavaa ei vielä todettu. Tulos viittaa siihen, että HR-perunat reagoivat herkemmin ympäristöön ja/tai kasvin kehitysasteeseen laukaisten puolustusvasteita, jotka saattavat parantaa kestävyyttä taudinaiheuttajia vastaan. Kolmas tutkittu kestävyystyyppi havaittiin Pito-perunalajikkeessa. Se muistutti nnr-kestävyyttä siten, että myös siinä viruksen liikkuminen nilassa muihin kasvinosiin estyi. PVA:n todettiin pysähtyvän vasta lehtiruodin tyvelle muodostuvaan irtoamisvyöhykkeeseen, mitä havainnollistettiin käyttämällä muunnettua PVA-rotua, joka tuotti UV-valossa fluoresoivaa vihreää valoa. Tulos viittaa siihen, että virus ei pääse kulkemaan vyöhykkeeseen kuuluvan suojaavan kerroksen läpi, jollei sillä ole pääsyä nilaan. Tällainen kestävyys on tarpeen, jotta virus ei voi korvata nilakuljetusta solusta soluun leviämisellä. Tulokset tuovat uusia näkökulmia kasvien viruskestävyyteen ja auttavat selittämään viruksen nilakuljetuksen estymistä sekä solusta soluun leviämisen pysähtymistä kestävissä kasveissa.

Integrin trafficking regulated by Rab21 is necessary for cytokinesis

Adherent cells undergo remarkable changes in shape during cell division. However, the functional interplay between cell adhesion turnover and the mitotic machinery is poorly understood. The endo/exocytic trafficking of integrins is regulated by the small GTPase Rab21, which associates with several integrin alpha subunits. Here, we show that targeted trafficking of integrins to and from the cleavage furrow is required for successful cytokinesis, and that this is regulated by Rab21. Rab21 activity, integrin-Rab21 association, and integrin endocytosis are all necessary for normal cytokinesis, which becomes impaired when integrin-mediated adhesion at the cleavage furrow fails. We also describe a chromosomal deletion and loss of Rab21 gene expression in human cancer, which leads to the accumulation of multinucleate cells. Importantly, reintroduction of Rab21 rescued this phenotype. In conclusion, Rab21-regulated integrin trafficking is essential for normal cell division, and its defects may contribute to multinucleation and genomic instability, which are hallmarks of cancer.

Kansalaisvaikuttamista? Habermasilaisen julkisuusteorian anti suomalaiselle ympäristöliikkeelle

Tässä pro gradu -tutkielmassa analysoidaan suomalaista ympäristöliikettä Jürgen Habermasin julkisuuden käsitteen näkökulmasta. Työn tutkimusongelmana on tarkastella julkisuutta ympäristövaikuttamisen keinona. Ponnahduslautana tutkimuksessa käytetään 2000-luvun Lapin metsien käyttöä koskevia kiistoja, joiden ratkomisyrityksiin moni Suomessa toimiva ympäristöjärjestö on osallistunut. Pitkään ratkaisemattomina olleet kiistat ovat johtaneet tilanteeseen, jossa on perusteltua kysyä, mikä ongelmien selvittämisessä on mennyt pieleen. Olisiko ennen kaikkea ammattimaiseen ja institutionalisoituneeseen vaikuttamiseen nojanneet ympäristöjärjestöt voineet toimia kiistojen yhteydessä jollakin vaihtoehtoisella tavalla? Vaihtoehdoksi esitetään Jürgen Habermasin tarjoamaa julkisuuden mallia, jonka on nähty olevan keskeinen modernin yhteiskunnan mahdollisuuksia ja rajoitteita arvioitaessa. Käsitys julkisuudesta sisältää ajatuksen kriittiseen keskusteluun pohjautuvasta, kaikille avoimesta kansalaisvaikuttamisesta. Näin muodostuneesta yleisestä mielipiteestä tulee poliittisen päätöksenteon ohjenuora. Julkisuuden ihanteen rinnalla Habermasin teoria sisältää kuvauksen julkisuuden rakennemuutoksesta – kriittisen julkisuuden alennustilasta myöhäiskapitalistisessa kulutusyhteiskunnassa. Tässä tutkimuksessa käytän näitä kahta teoreettista näkökulmaa arvioidessani nykyaikaista ympäristövaikuttamista. Keskeisin lähdeteos on Habermasin Julkisuuden rakennemuutos ja sen teoreettista perinnettä jatkaneet tutkimukset. Aineistoni koostuu kuudesta teemahaastattelusta, jotka toteutettiin keväällä 2009 Greenpeacessa, Suomen luonnonsuojeluliitossa ja WWF:ssä. Tutkimusmenetelmänä käytän teemoittelua. Sen tuloksena esiin nousevista teemoista keskeisin kertoo ympäristöjärjestöjen ammattimaistumisesta. Tulosteni mukaan ympäristövaikuttamista leimaa institutionalisoituneisuus ja hallinnollisuus, jolloin julkisuusperiaatteen toimintaedellytykset ovat heikentyneet. Julkisuuden käsitteelle löytyy kuitenkin myös varovaista tilausta suomalaisten ympäristöjärjestöjen keskuudessa.

Interrelations between Dry Eye Syndrome and Tear Fluid Phospholipid Transfer Protein

The simplified model of human tear fluid (TF) is a three-layered structure composed of a homogenous gel-like layer of hydrated mucins, an aqueous phase, and a lipid-rich outermost layer found in the tear-air interface. It is assumed that amphiphilic phospholipids are found adjacent to the aqueous-mucin layer and externally to this a layer composed of non-polar lipids face the tear-air interface. The lipid layer prevents evaporation of the TF and protects the eye, but excess accumulation of lipids may lead to drying of the corneal epithelium. Thus the lipid layer must be controlled and maintained by some molecular mechanisms. In the circulation, phospholipid transfer protein (PLTP) and cholesteryl ester transfer protein (CETP) mediate lipid transfers. The aim of this thesis was to investigate the presence and molecular mechanisms of lipid transfer proteins in human TF. The purpose was also to study the role of these proteins in the development of dry eye syndrome (DES). The presence of TF PLTP and CETP was studied by western blotting and mass spectrometry. The concentration of these proteins was determined by ELISA. The activities of the enzymes were determined by specific lipid transfer assays. To study the molecular mechanisms involved in PLTP mediated lipid transfer Langmuir monolayers and asymmetrical flow field-flow fractionation (AsFlFFF) was used. Ocular tissue samples were stained with monoclonal antibodies against PLTP to study the secretion route of PLTP. Heparin-Sepharose affinity chromatography was used for PLTP pull-down experiments and co-eluted proteins were identified with MALDI-TOF mass spectrometry or Western blot analysis. To study whether PLTP plays any functional role in TF PLTP-deficient mice were examined. The activity of PLTP was also studied in dry eye patients. PLTP is a component of normal human TF, whereas CETP is not. TF PLTP concentration was about 2-fold higher than that in human plasma. Inactivation of PLTP by heat treatment or immunoinhibition abolished the phospholipid transfer activity in tear fluid. PLTP was found to be secreted from lacrimal glands. PLTP seems to be surface active and is capable of accepting lipid molecules without the presence of lipid-protein complexes. The active movement of radioactively labeled lipids and high activity form of PLTP to acceptor particles suggested a shuttle model of PLTP-mediated lipid transfer. In this model, PLTP physically transports lipids between the donor and acceptor. Protein-protein interaction assays revealed ocular mucins as PLTP interaction partners in TF. In mice with a full deficiency of functional PLTP enhanced corneal epithelial damage, increased corneal permeability to carboxyfluorescein, and decreased corneal epithelial occludin expression was demonstrated. Increased tear fluid PLTP activity was observed among human DES patients. These results together suggest a scavenger property of TF PLTP: if the corneal epithelium is contaminated by hydrophobic material, PLTP could remove them and transport them to the superficial layer of the TF or, alternatively, transport them through the naso-lacrimal duct. Thus, PLTP might play an integral role in tear lipid trafficking and in the protection of the corneal epithelium. The increased PLTP activity in human DES patients suggests an ocular surface protective role for this lipid transfer protein.

Globalization and the Political : in the Borderlands with the Zapatista Movement

This study explores the meaning, content and significance of the political as manifest in the Mexican Zapatista movement as historically and geopolitically situated struggle. The case study undertakes a critical analysis of the development, organization, practice and discourse of the movement by drawing on fieldwork experiences, interviews, discussions, documents, films and other material produced by the movement, and the critical engagement with the research of others, especially in Latin America and Mexico. The dissertation poses the need to reconsider what constitutes and what we understand by the political , related particularly to the challenges provided by the critical globalization literature, decolonization and the study of social movements. The analysis encompasses several inter-related levels: the theoretical knowledge regarding the conceptualization of the political; the methodological level, regarding how such research can and should be conducted and knowledge claims formulated given the inescapable context and effects of global power relations; and the substantive level of adding specific information and analytical insights to existing knowledge of the Zapatista movement. As a result of conceptualization of a range of practices and processes, distinct understandings of the political can be underlined. Firstly, the conception of the indigenous and the struggles as indigenous movements as specifically political, not just a cultural or ethnic identity or a static quality but rather, an active consciousness integrally linked both to a longer history of oppression and as political articulation in the concrete context and lived experience of contemporary struggle. Secondly, the practice of autonomy as central to an understanding of the political in the context of the Zapatista struggle as a practical response to the situation of oppression, counter-insurgency, siege and conflict in Chiapas, as well as a positively informed mode of political self-understanding, expression and practice in its own right. Thirdly, the notion of geopolitical positioning as important to understanding of the political that encompasses the historicity of specific context and the power relations which shape that context, developed in two different ways: in regard to the positioning of the researcher and knowledge production with and about the Zapatistas, and in regard to the practice and knowledge of the Zapatistas as a decolonizing force in their encounters, interaction and relations with others, especially the global civil society. Finally, the role of silence, absence, invisibility, revelation and hiding in political practice as a deliberate strategy in response to oppressive power. -

Pre-eclampsia : The role of soluble VEGF receptor-1 and related anti-angiogenic factors beyond

Pre-eclampsia is a pregnancy complication that affects about 5% of all pregnancies. It is known to be associated with alterations in angiogenesis -related factors, such as vascular endothelial growth factor (VEGF). An excess of antiangiogenic substances, especially the soluble receptor-1 of VEGF (sVEGFR-1), has been observed in maternal circulation after the onset of the disease, probably reflecting their increased placental production. Smoking reduces circulating concentrations of sVEGFR-1 in non-pregnant women, and in pregnant women it reduces the risk of pre-eclampsia. Soluble VEGFR-1 acts as a natural antagonist of VEGF and placental growth factor (PlGF) in human circulation, holding a promise for potential therapeutic use. In fact, it has been used as a model to generate a fusion protein, VEGF Trap , which has been found effective in anti-angiogenic treatment of certain tumors and ocular diseases. In the present study, we evaluated the potential use of maternal serum sVEGFR-1, Angiopoietin-2 (Ang-2) and endostatin, three central anti-angiogenic markers, in early prediction of subsequent pre-eclampsia. We also studied whether smoking affects circulating sVEGFR-1 concentrations in pregnant women or their first trimester placental secretion and expression in vitro. Last, in order to allow future discussion on the potential therapy based on sVEGFR-1, we determined the biological half-life of endogenous sVEGFR-1 in human circulation, and measured the concomitant changes in free VEGF concentrations. Blood or placental samples were collected from a total of 268 pregnant women between the years 2001 2007 in Helsinki University Central Hospital for the purposes above. The biomarkers were measured using commercially available enzyme-linked immunosorbent assays (ELISA). For the analyses of sVEGFR-1, Ang-2 and endostatin, a total of 3 240 pregnant women in the Helsinki area were admitted to blood sample collection during two routine ultrasoundscreening visits at 13.7 ± 0.5 (mean ± SD) and 19.2 ± 0.6 weeks of gestation. Of them, 49 women later developing pre-eclampsia were included in the study. Their disease was further classified as mild in 29 and severe in 20 patients. Isolated early-onset intrauterine growth retardation (IUGR) was diagnosed in 16 women with otherwise normal medical histories and uncomplicated pregnancies. Fifty-nine women remaining normotensive, non-proteinuric and finally giving birth to normal-weight infants were picked to serve as the control population of the study. Maternal serum concentrations of Ang-2, endostatin and sVEGFR-1, were increased already at 16 20 weeks of pregnancy, about 13 weeks before the clinical manifestation of preeclampsia. In addition, these biomarkers could be used to identify women at risk with a moderate precision. However, larger patient series are needed to determine whether these markers could be applied for clinical use to predict preeclampsia. Intrauterine growth retardation (IUGR), especially if noted at early stages of pregnancy and not secondary to any other pregnancy complication, has been suggested to be a form of preeclampsia compromising only the placental sufficiency and the fetus, but not affecting the maternal endothelium. In fact, IUGR and preeclampsia have been proposed to share a common vascular etiology in which factors regulating early placental angiogenesis are likely to play a central role. Thus, these factors have been suggested to be involved in the pathogenesis of IUGR. However, circulating sVEGFR-1, Ang-2 and endostatin concentrations were unaffected by subsequent IUGR at early second trimester. Furthermore, smoking was not associated with alterations in maternal circulating sVEGFR-1 or its placental production. The elimination of endogenous sVEGFR-1 after pregnancy was calculated from serial samples of eight pregnant women undergoing elective Caesarean section. As typical for proteins in human compartments, the elimination of sVEGFR-1 was biphasic, containing a rapid halflife of 3.4 h and a slow one of 29 h. The decline in sVEGFR-1 concentrations after mid-trimester legal termination of pregnancy was accompanied with a simultaneous increase in the serum levels of free VEGF so that within a few days after pregnancy VEGF dominated in the maternal circulation. Our study provides novel information on the kinetics of endogenous sVEGFR-1, which serves as a potential tool in the development of new strategies against diseases associated with angiogenic imbalance and alterations in VEGF signaling.

Tsoi Yuen Resistance and Post-80s Generation: A new wave of domestic social movement

Hong Kong was once a British colony and has been under the sovereignty of People’s Republic of China (PRC) since 1997. However, some of the unjust practices and colonial legacies are infiltrated into the development ideology as well as the social structures. The construction of intercity express railway project announced in 2008 causing the demolishment of Tsoi Yuen Tsuen, a “non-indigenous” agricultural village in Hong Kong, was one of the current examples. Tsoi Yuen village was established under the former colonial sovereignty sixty years ago. Approximately 450 populations were affected that they had to relocate their homeland involuntarily. However, these villagers were very attached to their homelands and were unwilling to move, and meanwhile they found that they were absent in the government’s consultation and decision-making process. Soon they began their resistance and demanded for “No Move! No Demolish!”. Their movement was strongly supported by a group of “Post-80s generation” and turned into the most important social movement of the city in recent years. In fact, demolition of Tsoi Yuen Village for city development is not an isolated case in the city. Meanwhile the situation is getting worse in Mainland China. I chose the case study of Tsoi Yuen Resistance from 2008 to 2011 for revelation of the complicated colonial history and postcolonial era of Hong Kong. I focused on discussing the Tsoi Yuen Resistance and the Post-80s movement, and how they have exposed the tension between top-down urban planning and development and public movements fighting for a more democratic process in choosing their way of living. Through the study of a village movement which as well as the rationale behind the Post-80s’ support, I hoped to illustrate how this movement has awaken a different sense of living for the new generations in the midst of the high-sounding urban development. It is an opportunity to examine Hong Kong’s colonial epoch in a different perspective: through studying the Tsoi Yuen Village, let them (subalterns) speak for themselves. Furthermore, the significance of this resistance, taking place eleven years after the handover to the PRC, is an important fact that I shall not miss in later discussion. Last but not least, during the resistance, advanced technology and social networks such as Facebook, Twitter, iPhone were used by Post 80s generation to spread the latest information in order to attract public’s concern and participation. Therefore, apart from studying Tsoi Yuen Resistance as a local social movement, I also regard it as a part of the global movement in perusing ecological lifestyle and civil society. How Post 80s’ generation manipulates the global idea in a local context will also be examined.

Tracing water movement by means of Escherichia coli bacteriophages

Tiivistelmä: Escherichia coli bacteriofaagit merkkiaineena vesien kulkeutumistutkimuksissa