Tumor-suppressor gene promoter hypermethylation in saliva of head and neck cancer patients

Head and neck squamous cell carcinoma (HNSCC) accounts for a bulk of the oral and laryngeal cancers, the majority (70%) of which are associated with smoking and excessive drinking, major known risk factors for the development of HNSCC. In contrast to reports that suggest an inverse relationship between smoking and global DNA CpG methylation, hypermethylation of promoters of a number of genes was detected in saliva collected from patients with HNSCC. Using a sensitive methylation-specific polymerase chain reaction (MSP) assay to determine specific methylation events in the promoters of RASSF1A, DAPK1, and p16 genes, we demonstrate that we can detect tumor presence with an overall accuracy of 81% in the DNA isolated from saliva of patients with HNSCC (n = 143) when compared with the DNA isolated from the saliva of healthy nonsmoker controls (n = 31). The specificity for this MSP panel was 87% and the sensitivity was 80%(with a Fisher exact test P < .0001). In addition, the test panel performed extremely well in the detection of the early stages of HNSCCs, with a sensitivity of 94% and a specificity of 87%, and a high. concordance value of 0.8, indicating an excellent overall agreement between the presence of HNSCC and a positive MSP panel result. In conclusion, we demonstrate that the promoter methylation of RASSF1A, DAPK1, and p16 MSP panel is useful in detecting hypermethylation events in a noninvasive manner in patients with HNSCC.

The tumour-promoting receptor tyrosine kinase, EphB4, regulates expression of Integrin-β8 in prostate cancer cells

Background The EphB4 receptor tyrosine kinase is overexpressed in many cancers including prostate cancer. The molecular mechanisms by which this ephrin receptor influences cancer progression are complex as there are tumor-promoting ligand-independent mechanisms in place as well as ligand-dependent tumor suppressive pathways. Methods We employed transient knockdown of EPHB4 in prostate cancer cells, coupled with gene microarray analysis, to identify genes that were regulated by EPHB4 and may represent linked tumor-promoting factors. We validated target genes using qRT-PCR and employed functional assays to determine their role in prostate cancer migration and invasion. Results We discovered that over 500 genes were deregulated upon EPHB4 siRNA knockdown, with integrin β8 (ITGB8) being the top hit (29-fold down-regulated compared to negative non-silencing siRNA). Gene ontology analysis found that the process of cell adhesion was highly deregulated and two other integrin genes, ITGA3 and ITGA10, were also differentially expressed. In parallel, we also discovered that over-expression of EPHB4 led to a concomitant increase in ITGB8 expression. In silico analysis of a prostate cancer progression microarray publically available in the Oncomine database showed that both EPHB4 and ITGB8 are highly expressed in prostatic intraepithelial neoplasia, the precursor to prostate cancer. Knockdown of ITGB8 in PC-3 and 22Rv1 prostate cancer cells in vitro resulted in significant reduction of cell migration and invasion. Conclusions These results reveal that EphB4 regulates integrin β8 expression and that integrin β8 plays a hitherto unrecognized role in the motility of prostate cancer cells and thus targeting integrin β8 may be a new treatment strategy for prostate cancer.

Osobennosti nacional'noj ohoty na pisatelja : Literaturnaja bor'ba 2002 goda vokrug romana Vladimira Sorokina Goluboe salo

Tutkielman lähtökohtana on oikeusjuttu, joka Venäjällä nostettiin kirjailija Vladimir Sorokinia vastaan kesällä 2002. Sorokinia ja hänen kustantamoaan Ad Marginemia syytettiin pornografian levittämisestä romaanissa Goluboe salo (1999), tarkemmin sanoen sivuilla 256 262, joilla kuvataan kommunistijohtajien Stalinin ja Hruščëvin homoseksuaalinen akti. Syytteen takana oli konservatiivinen, presidentti Vladimir Putinia ihannoiva nuorisoliike Iduščie vmeste ("Yhdessäkulkijat"). Oikeusjuttu ja siihen liittyneet tapahtumat - prosessia kutsuttiin venäläisissä tiedotusvälineissä yksinkertaisesti "Sorokinin jutuksi" ("delo Sorokina") - nostattivat Venäjällä laajan ja vilkkaan ns. kirjasodan kaunokirjallisuuden moraalitehtävästä ja kirjailijan vastuusta/vapaudesta. Työssä selvitellään, miksi nimenomaan kirjailija Vladimir Sorokin ja nimenomaan romaani Goluboe salo aiheuttivat skandaalin, mitä argumentteja polemiikissa esiintyi ja miten tapahtumat heijastavat Venäjän yhteiskunnallista tilannetta, etenkin kaunokirjallisuuden nykyistä asemaa. Metodologisesti tutkielma sijoittuu kirjallisuussosiologian ja reseptiotutkimuksen (vastaanottotutkimuksen) alalle. Tutkielman aineisto - 71 lehtiartikkelia - on koottu 31 venäläisestä valtakunnallisesta sanoma- ja aikakauslehdestä Integrum-tietokantaa hyväksikäyttäen. "Sorokinin juttua" analysoidaan artikkeleiden valossa kolmesta näkökulmasta: suhteessa 1) seksuaalisuuden ja 2) kaunokirjallisuuden asemaan Venäjällä sekä lyhyemmin suhteessa 3) muutamiin yleisyhteiskunnallisiin aspekteihin. Taustaksi selvitellään venäläisen postmodernismin teoriaa ja nk. kirjallisuusinstituution toimintaa. Tutkielmassa esitetään, että skandaali kertoo osaltaan niistä ongelmista, joiden kanssa Venäjä 2000-luvulla joutuu painiskelemaan. "Sorokinin juttu" laajeni pitkälti yli perinteisen pornografiakysymyksen; kirjasodassa tuli tiedotusvälineissä käsiteltyä seksuaalisuuden ja kaunokirjallisuuden aseman lisäksi mm. sananvapautta, tapauksen poliittisia konnotaatioita, "älymystön" ja "kansan" suhdetta, neuvostomenneisyyden pimeitä puolia sekä Venäjän suhdetta länteen. Jupakka myös nähtiin erinomaisena PR:nä Sorokinille. Goluboe salo näyttää joutuneen oikeusprosessin kohteeksi, koska siinä a) tuodaan venäläiseen kaunokirjallisuuteen avoin ja yksityiskohtainen seksuaalisuuden ja ruumiintoimintojen kuvaus b) herjataan häpeämättömästi venäläisen kaunokirjallisuuden kanonisoituja klassikoita sekä kritisoidaan terävästi neuvostomenneisyyttä, jota Venäjä ei ole vielä kyennyt tyhjentävästi käsittelemään. Skandaalissa asettuivat vastakkain ns. traditionalistinen ja postmodernistinen kirjallisuuskäsitys; edellisen mukaan kirjailijoiden tulisi kantaa yhteiskunnallinen vastuunsa, jälkimmäinen tahtoo nähdä kaunokirjallisuuden itsenäisempänä ilmiönä. Tutkimuksen lopputuloksena valtaenemmistö aineistosta näyttää enemmän tai vähemmän puolustavan Sorokinia - tosin ei niinkään hänen romaaninsa kirjallisten ansioiden vuoksi (joita harvat kiittivät), vaan silkasta periaatteesta: sanan- ja valinnanvapautta ei tahdota nähdä enää kahlittavan. Avainsanat: Sorokin - pornografia - kaunokirjallisuus - kirjallisuussosiologia - vastaanotto

Liivin kielen ajan perusyksiköihin perustuvat ajan adverbiaalit

Pro gradu –työni tutkimuskohteena ovat liivin kielen ajan adverbiaalit, jotka on muodostettu ajan perusyksiköiden pohjalta. Tutkimus on aineistolähtöinen ja perustuu 563:een esimerkkilauseeseen. Ne on koottu kolmesta kielennäytekokoelmasta: E. N. Setälän ja Väinö Kyrölän kokoelmasta Näytteitä liivin kielestä (1953), Seppo Suhosen kokoelmasta Liivin kielen näytteitä (1975) sekä Julius Mägisten kokoelmasta Muistoja Liivinrannasta (2006). Lisäksi esimerkkejä on Lauri Kettusen liivin sanakirjasta Livisches Wörterbuch (1938). Käsittelemäni ajan perusyksiköt ovat aikaa nimeäviä substantiiveja, kuten vuorokaudenaikojen, viikonpäivien, kuukausien ja vuodenaikojen nimitykset sekä merkitykset ’vuosi’, ’kuukausi’, ’viikko’, ’tunti’, ’minuutti’ ja ’sekunti’. Liivin kielessä näistä muodostetaan ajan adverbiaaleja leksikaalisin ja syntaktisin keinoin. Leksikaaliset ajan adverbiaalit rajautuvat semanttisin perustein: ne ovat sijamuodoissa taivutettuja substantiiveja. Syntaktiset ajan adverbiaalit ovat useampisanaisia adpositiolausekkeita ja kiinteitä sanaliittoja. Ne on muodostettu adpositioiden, kvanttoripronominien, demonstratiivipronominien tai muiden määritteiden avulla ajan perusyksiköiden pohjalta. Tutkielmassani käsittelen aikaa myös hieman laajemmasta näkökulmasta: käsittelen aikaa ilmaisevien yksiköiden muodostumista sekä ajan kielellistämistä. Esittelen myös aiempaa tutkimusta ajan adverbiaaleista sekä tähän mennessä laaditut liivin kieliopin kuvaukset. Ne ovat keskittyneet enemmän morfologian kuin syntaksin esittelyyn. Ajan adverbiaalit ovat jääneet vain maininnan asteelle, joten käsittelemäni aihe on varsin tutkimatonta aluetta liivin kielessä. Adverbiaalien ja tarkemmin ajan adverbiaalien määrittelyn olen perustanut Ison suomen kieliopin pohjalle. Yleisestikin läpi työni kulkee liivin kielen rakenteiden ja ilmiöiden vertailu erityisesti suomeen ja viroon. Taustaksi tutkimukselleni esittelen ajan perusyksiköt ja niiden etymologiaa Itämerenalueen kielistä suomessa, virossa, saksassa, ruotsissa, latviassa ja venäjässä. Liivin aineiston käsittely alkaa myös ajan perusyksiköiden ja niistä muodostettujen leksikaalisten ajan adverbiaalien esittelyllä. Leksikaaliset ajan adverbiaalit muodostetaan liivissä usean eri sijamuodon avulla. Yleisin on partitiivi-illatiivi. Muut käytetyt sijat ovat nominatiivi-genetiivi, illatiivi, inessiivi ja adessiivi sekä epäproduktiivinen essiivi. Leksikaalisten ajan adverbiaalien yhteydessä viittaan myös objektin sijaiseen määrän adverbiaaliin eli osmaan. Syntaktisten ajan adverbiaalien yhteydessä käsittelen sekä niiden muodostamista yleensä että niitä muodostavien elementtien muotoa. Adpositiolausekkeiden yhteydessä käsittelen myös adpositioiden ilmenemistä itämerensuomalaisissa kielissä. Aineistossani esiintyy 11 eri adpositiota tai adposition tavoin käytettyä nominia. Kuvaan sekä adpositioiden itsensä muotoa että adpositioiden täydennyksen muotoa. Ajan adverbiaaleja muodostavia kvanttoripronomineja esiintyy aineistossani viisi. Osa niistä ajan adverbiaaleina esiintyessään taipuu ja usein myös kongruoi täydennyksensä kanssa, osa on taipumattomia ja niiden täydennys on lähes aina nominatiivi-genetiivissä. Ajan adverbiaaleja muodostetaan myös kolmen demonstratiivipronominin avulla. Demonstratiivipronomini ja sen pääsana voivat kongruoida tai pääsana voi esiintyä samassa muodossa kuin yksin leksikaalisena ajan adverbiaalina. Muita määritteitä aineistossani on neljä. Kolme niistä on nomineja ja yksi on partisiippi. Ne eivät useimmiten itse taivu ja niiden täydennys on tavallisesti nominatiivi-genetiivissä, mutta joissakin tapauksissa myös paikallissijoissa. Työssäni olen viitannut myös leksikaalistumiseen. Liivissä useat muissa itämerensuomalaisissa kielissä esiintyvät sijamuodot ovat muuttuneet epäproduktiivisiksi. Tällaisia ovat ulkopaikallissijat sekä essiivi ja instruktiivi. Näitä kaikkia kuitenkin esiintyy ajan ilmausten yhteydessä ja ulkopaikallissijoista erityisesti adessiivilla näyttäisi olevan tärkeä asema sekä ajan että paikan ilmauksissa.

Guaiac versus immunochemical tests: Faecal occult blood test screening for colorectal cancer in a rural community

Objective: To describe patient participation and clinical performance in a colorectal cancer (CRC) screening program utilising faecal occult blood test (FOBT). Methods: A community-based intervention was conducted in a small, rural community in north Queensland, 2000/01. One of two FOBT kits – guaiac (Hemoccult-ll) or immunochemical (Inform) – was assigned by general practice and mailed to participants (3,358 patients aged 50–74 years listed with the local practices). Results: Overall participation in FOBT screening was 36.3%. Participation was higher with the immunochemical kit than the guaiac kit (OR=1.9, 95% Cl 1.6-2.2). Women were more likely to comply with testing than men (OR=1.4, 95% Cl 1.2-1.7), and people in their 60s were less likely to participate than those 70–74 years (OR=0.8, 95% Cl 0.6-0.9). The positivity rate was higher for the immunochemical (9.5%) than the guaiac (3.9%) test (χ2=9.2, p=0.002), with positive predictive values for cancer or adenoma of advanced pathology of 37.8% (95% Cl 28.1–48.6) for !nform and 40.0% (95% Cl 16.8–68.7) for Hemoccult-ll. Colonoscopy follow-up was 94.8% with a medical complication rate of 2–3%. Conclusions: An immunochemical FOBT enhanced participation. Higher positivity rates for this kit did not translate into higher false-positive rates, and both test types resulted in a high yield of neoplasia. Implications: In addition to type of FOBT, the ultimate success of a population-based screening program for CRC using FOBT will depend on appropriate education of health professionals and the public as well as significant investment in medical infrastructure for colonoscopy follow-up.

Genetic Basis of Pituitary Adenoma Predisposition

Much of the global cancer research is focused on the most prevalent tumors; yet, less common tumor types warrant investigation, since A rare disorder is not necessarily an unimportant one . The present work discusses a rare tumor type, the benign adenomas of the pituitary gland, and presents the advances which, during the course of this thesis work, contributed to the elucidation of a fraction of their genetic background. Pituitary adenomas are benign neoplasms of the anterior pituitary lobe, accounting for approximately 15% of all intracranial tumors. Pituitary adenoma cells hypersecrete the hormones normally produced by the anterior pituitary tissue, such as growth hormone (GH) and prolactin (PRL). Despite their non-metastasizing nature, these adenomas can cause significant morbidity and have to be adequately treated; otherwise, they can compromise the patient s quality of life, due to conditions provoked by hormonal hypersecretion, such as acromegaly in the case of GH-secreting adenomas, or due to compressive effects to surrounding tissues. The vast majority of pituitary adenomas arise sporadically, whereas a small subset occur as component of familial endocrine-related tumor syndromes, such as Multiple Endocrine Neoplasia type 1 (MEN1) and Carney complex (CNC). MEN1 is caused by germline mutations in the MEN1 tumor suppressor gene (11q13), whereas the majority of CNC cases carry germline mutations in the PRKAR1A gene (17q24). Pituitary adenomas are also encountered in familial settings outside the context of MEN1 and CNC, but unlike in the latter syndromes, their genetic background until recently remained elusive. Evidence in previous literature supported the notion that a tumor suppressor gene on 11q13, residing very close to but still distinct from MEN1, causes genetic susceptibility to pituitary tumors. The aim of the study was to identify the genetic cause of a low penetrance form of Pituitary Adenoma Predisposition (PAP) in families from Northern Finland. The present work describes the methodological approach that led to the identification of aryl hydrocarbon receptor interacting protein (AIP) as the gene causing PAP. Combining chip-based technologies (SNP and gene expression arrays) with traditional gene mapping methods and genealogy data, we showed that germline AIP mutations cause PAP in familial and sporadic settings. PAP patients were diagnosed with mostly adenomas of the GH/PRL-secreting cell lineage. In Finland, two AIP mutations accounted for 16% of all patients diagnosed with GH-secreting adenomas, and for 40% of patients being younger than 35 years of age at diagnosis. AIP is suggested to act as a tumor suppressor gene, a notion supported by the nature of the identified mutations (most are truncating) and the biallelic inactivation of AIP in the tumors studied. AIP has been best characterized as a cytoplasmic interaction partner of aryl hydrocarbon receptor (AHR), also known as dioxin receptor, but it has other partners as well. The mechanisms that underlie AIP-mediated pituitary tumorigenesis are to date largely unknown and warrant further investigation. Because AIP was identified in the genetically homogeneous Finnish population, it was relevant to examine its contribution to PAP in other, more heterogeneous, populations. Analysis of pituitary adenoma patient series of various ethnic origins and differing clinical settings revealed germline AIP mutations in all cohorts studied, albeit with low frequencies (range 0.8-7.4%). Overall, PAP patients were typically diagnosed at a young age (range 8-41 years), mainly with GH-secreting adenomas, without strong family history of endocrine disease. Because many PAP patients did not display family history of pituitary adenomas, detection of the condition appeared challenging. AIP immunohistochemistry was tested as a molecular pre-screening tool on mutation-positive versus mutation-negative tumors, and proved to be a potentially useful predictor of PAP. Mutation screening of a large cohort of colorectal, breast, and prostate tumors did not reveal somatic AIP mutations. These tumors, apart from being the most prevalent among men and women worldwide, have been associated with acromegaly, particularly colorectal neoplasia. In this material, AIP did not appear to contribute to the pathogenesis of these common tumor types and other genes seem likely to play a role in such tumorigenesis. Finally, the contribution of AIP in pediatric onset pituitary adenomas was examined in a unique population-based cohort of sporadic pituitary adenoma patients from Italy. Germline AIP mutations may account for a subset of pediatric onset GH-secreting adenomas (in this study one of seven GH-secreting adenoma cases or 14.3%), and appear to be enriched among young (≤25 years old) patients. In summary, this work reveals a novel tumor susceptibility gene, namely AIP, which causes genetic predisposition to pituitary adenomas, in particular GH-secreting adenomas. Moreover, it provides molecular tools for identification of individuals predisposed for PAP. Further elaborate studies addressing the functional role of AIP in normal and tumor cells will hopefully expand our knowledge on endocrine neoplasia and reveal novel cellular mechanisms of pituitary tumorigenesis, including potential drug targets.

Molecular basis of colorectal cancer predisposition

Colorectal cancer (CRC) is one of the most frequent malignancies in Western countries. Inherited factors have been suggested to be involved in 35% of CRCs. The hereditary CRC syndromes explain only ~6% of all CRCs, indicating that a large proportion of the inherited susceptibility is still unexplained. Much of the remaining genetic predisposition for CRC is probably due to undiscovered low-penetrance variations. This study was conducted to identify germline and somatic changes that contribute to CRC predisposition and tumorigenesis. MLH1 and MSH2, that underlie Hereditary non-polyposis colorectal cancer (HNPCC) are considered to be tumor suppressor genes; the first hit is inherited in the germline and somatic inactivation of the wild type allele is required for tumor initiation. In a recent study, frequent loss of the mutant allele in HNPCC tumors was detected and a new model, arguing against the two-hit hypothesis, was proposed for somatic HNPCC tumorigenesis. We tested this hypothesis by conducting LOH analysis on 25 colorectal HNPCC tumors with a known germline mutation in the MLH1 or MSH2 genes. LOH was detected in 56% of the tumors. All the losses targeted the wild type allele supporting the classical two-hit model for HNPCC tumorigenesis. The variants 3020insC, R702W and G908R in NOD2 predispose to Crohn s disease. Contribution of NOD2 to CRC predisposition has been examined in several case-control series, with conflicting results. We have previously shown that 3020insC does not predispose to CRC in Finnish CRC patients. To expand our previous study the variants R702W and G908R were genotyped in a population-based series of 1042 Finnish CRC patients and 508 healthy controls. Association analyses did not show significant evidence for association of the variants with CRC. Single nucleotide polymorphism (SNP) rs6983267 at chromosome 8q24 was the first CRC susceptibility variant identified through genome-wide association studies. To characterize the role of rs6983267 in CRC predisposition in the Finnish population, we genotyped the SNP in the case-control material of 1042 cases and 1012 controls and showed that G allele of rs6983267 is associated with the increased risk of CRC (OR 1.22; P=0.0018). Examination of allelic imbalance in the tumors heterozygous for rs6983267 revealed that copy number increase affected 22% of the tumors and interestingly, it favored the G allele. By utilizing a computer algorithm, Enhancer Element Locator (EEL), an evolutionary conserved regulatory motif containing rs6983267 was identified. The SNP affected the binding site of TCF4, a transcription factor that mediates Wnt signaling in cells, and has proven to be crucial in colorectal neoplasia. The preferential binding of TCF4 to the risk allele G was showed in vitro and in vivo. The element drove lacZ marker gene expression in mouse embryos in a pattern that is consistent with genes regulated by the Wnt signaling pathway. These results suggest that rs6983267 at 8q24 exerts its effect in CRC predisposition by regulating gene expression. The most obvious target gene for the enhancer element is MYC, residing ~335 kb downstream, however further studies are required to establish the transcriptional target(s) of the predicted enhancer element.

Mode of action of antitumour antibiotics: Spectrophotometric studies on the interaction of chromomycin A3 with DNA and chromatin of normal and neoplastic tissue

The binding of chromomycin A3, an antitumour antibiotic, to various DNA and chromatin isolated from mouse and rat liver, mouse fibrosarcoma and Yoshida ascites sarcoma cells was studied spectrophotometrically at 29°C in 10−2 M Tris-HCl buffer, pH 8.0, containing small amounts of MgCl2 (4.5 · 10−5−25 · 10−5 M). An isobestic point at 415 nm was observed when chromomycin A3 was gradually titrated with Image and its spectrum shifted towards higher wavelength. The rates and extent of these spectral changes were found to be dependent on the concentration of Mg2+. The change in absorbance at 440 nm was used to calculate apparent binding constant (Ka p M−1) and sites per nucleotide (n) from Scatchard plots for various DNA and chromatins. As expected, values of n for chromatin (0.06–0.10) were found to be lower than that found for corresponding DNA (0.10–0.15). Apparently no such correlation exists between binding constants (Ka p M−1 · 10−4) of DNA (6.4–11.2) and of chromatin (3.1–8.3), but Ka p M−1 of chromatin isolated from mouse fibrosarcoma and Yoshida ascites sarcoma are 1.5–3 times higher than that found for mouse and rat liver chromatin. These differences may be taken to indicate structural difference in nucleoprotein complexes caused by neoplasia. The relevance of this finding to tumour suppressive action of chromomycin A3 is discussed.

Valoa itään? : Kansanlähetys ja Neuvostoliitto 1967-1973

Light to the East? The Finnish Lutheran Mission and the Soviet Union 1967 1973 The Cold War affected the lives of Christian churches, especially in Europe. Besides the official ecumenical relations between east and west, there existed unofficial activity from west to east, such as smuggling Bibles and distributing information about the severe condition of human rights in the USSR. This study examines this kind of unofficial activity originating in Finland. It especially concentrates on the missionary work to the Soviet Union done by the Finnish Lutheran Mission (FLM, Suomen Evankelisluterilainen Kansanlähetys) founded in 1967. The work for Eastern Europe was organised through the Department for the Slavic Missions. FLM was founded within the Evangelical Lutheran Church of Finland, but it was not connected to the church on an organisational level. In addition to the strong emphasis on the Lutheran confession, FLM presented evangelical theology. The fundamental work of the Department for the Slavic Missions was to organise the smuggling of Bibles and other Christian literature to the Soviet Union and other countries behind the iron curtain. They also financed several Christian radio programmes produced and aired mainly by the international Trans World Radio. The Department diversified its activity to humanitarian help by distributing material help such as clothes and shoes to the unregistered evangelical and baptist groups, which were called the underground churches . In Finland the Department focused on information services. It published its own magazine, Valoa idässä (Light in the East), 5 to 6 times per year. Through the magazine and by distributing samizdat material received from the unregistered Christian groups, it discussed and reported the violations of human rights in the Soviet Union, especially when the unregistered Christian groups were considered the victims. The resistance against the Soviet Union was not as much political but religious: the staff of the Department were religious and revivalist young people who thought, for instance, that communism was in some way an apocalyptic world power revealed in the Bible. Smuggling Bibles was discussed widely in the Finnish media and even in parliament and the Finnish Security Police (SUPO, Suojelupoliisi) and in the Lutheran Church. From the church s point of view, this kind of missionary work was understandable but bothersome. Through their ecumenical connections, the bishops knew the critical situation of churches behind the iron curtain very well, but wanted to act diplomatically and cautiously to prevent causing harm to ecumenical or political relations. The leftist media and members of parliament especially accused the work of the Department of being illegal and endangering relations between Finland and the Soviet Union. SUPO did not consider the work of the Department as illegal activity or as a threat to Finnish national security.

"Mikä ihana kuva: isä, äiti ja lapset Herran työssä" : Lähetystyöntekijöiden perhe-elämä Kiinassa 1910- ja 1920-luvuilla

Tutkimukseni käsittelee Suomen Lähetysseuran Kiinan-työssä olleita perheitä vuosina 1915–1928. Enemmistö tuolloin SLS:n lähetystössä olleista läheteistä oli perheellisiä, ja perheiden elämäntilanteet lähetyskentällä vaikuttivat koko SLS:n yhteisön työmahdollisuuksiin. Tutkimukseni kohdehenkilöt ovat Signe ja Väinö Kantele sekä Inkeri ja Toivo Koskikallio perheineen, ja he edustavat sitä kokonaisuutta, jonka perheelliset lähetit Kiinassa muodostivat. Tutkin pro gradu -työssäni, millaista lähetystyöntekijöiden perhe-elämä oli lähetyskentällä. Kysyn myös, miten lähetystyö vaikutti perheeseen, ja miten perhe vaikutti lähetystyön tekemiseen. Lisäksi selvitän, miten kiinalainen kulttuuri vaikutti lähettiperheiden elämään. Tutkimukseni tärkeimmät lähteet ovat Signe Kanteleen kirjeet omaisilleen sekä Inkeri Koskikallion kirjeet ja päiväkirjat. Lähetyshistoriaa ei ole aiemmin tutkittu perheen näkökulmasta, joten tutkimus on hyvin aineistolähtöinen. Tärkeimpään käyttämääni kirjallisuuteen kuuluvat SLS:n Kiinan-työn historiaan sekä naislähetystyöntekijöihin liittyvät tutkimukset. Lähetystyöntekijät solmivat Kiinassa keskenään useita avioliittoja. Jotkut läheteistä olivat avioituneet jo Suomessa. SLS:n johtokunta kontrolloi lähettien avioliittoja ja myös perheiden lapsia, joten perhe ei ollut pelkästään lähettien yksityisasia. 1910- ja 1920-luvut olivat erityisen lapsirikasta aikaa, mikä vaikutti merkittävästi SLS:n Kiinan-yhteisön toimintaan. Joihinkin perheisiin oli syntynyt lapsia jo Suomessa ja lähetyskentällä perheisiin syntyi tutkimusajankohtana 26 lasta. Monet lähettiperheiden haasteista liittyivät lähetystyön ja perhe-elämän yhdistämiseen. Perheenäideillä oli vahva lähetyskutsumus, mutta raskaudet, synnytykset ja elämä pienten lasten kanssa rajoittivat naisten työskentelymahdollisuuksia ja aiheuttivat rooliristiriitoja. Perheen arjessa haasteena oli myös perheenisän poissaolo lähetystyöhön liittyneiden matkojen takia. Erityisesti tuolloin korostui lähetysasemalla asuneiden muiden suomalaisten läsnäolon tärkeys, vaikka SLS:n yhteisön tiiviys myös rajoitti perheiden yksityisyyttä. Perhe-elämällä oli SLS:n tuki, sillä monien muiden protestanttisten lähetysseurojen tavoin SLS kannusti perheellisiä lähettejään hyödyntämään perhettään evankelioimistyössä ja toimimaan kristityn perheen esimerkkinä paikallisille. Perheen esimerkillisyyteen kannustamisesta huolimatta Suomen Lähetysseura oli virallisissa julkaisuissaan vaitonainen perhetapahtumien vaikutuksesta työhön lähetyskentällä. Työn tukijat haluttiin vakuuttaa työn häiriöttömyydestä, vaikka todellisuus Kiinassa oli ajoittain toinen. Perheenäitien ja lasten sairastelut pakottivat perheenisiä välillä vähentämään tai lopettamaan työskentelyä. Lähetit myös menettivät Kiinassa lapsia ja puolisoita, mikä luonnollisesti vaikutti lähettiyhteisön toimintaan. Suomalaisvanhemmat kokivat perhe-elämän kiinalaisen kulttuurin keskellä ajoittain haasteelliseksi. Ristiriitatilanteita aiheuttivat kiinalaisten erilaiset elintavat ja esimerkiksi suomalaisten mielestä kyseenalainen lastenhoito- ja kasvatuskulttuuri. Suomalaisperheiden elämä Kiinassa oli joiltain osin samanlaista kuin heidän aikakautenaan Suomessa, mutta lähetystyö ja kiinalainen kulttuuri toivat siihen oman erikoisleimansa. Elämä perheenä lähetystyössä sisälsi paljon ulkoapäin tulleita ja sisäisiä haasteita. Lähetit kertoivat näistä haasteista, mutta korostivat myös vahvaa hengellistä kutsumustaan, tyytyväisyyttään elämäänsä sekä perheensä onnellisuutta.

Herrat, jätkät ja sotataito : Kansalaissotilas- ja ammattisotilasarmeijan rakentuminen 1920- ja 1930-luvulla

Gentlemen, Lads and the Art of War The Construction of Citizen Soldier- and Professional Soldier Armies into the Miracle of the Winter War During the 1920s and 1930s The Miracle of the Winter War was not a myth - at least according to them, who were making that miracle to happen. This study is not just about the Armed Forces and society, but moreover a study about civil society inside the organization of armed forces. Conscription kept Finnish military organization (and is still keeping) very closely connected with civil society and therefore there is no need to locate the possible critical misunderstandings brought by two different identity-based approaches. The great performance of the Armed Forces during the Second World War was not made of superior art of war. It was not the high level of discipline either. Art of war is basically a (deep level) cultural level equation that has more to do with culturally absorbed schemes of meaning making than rational decision-making. Naturally attrition based approach to effect-making directed the organizational methods in attrition based organisational practices, where there were only minor possibilities to practice any manoeuvre-based organisational behaviour. The practice and method of leadership lent similarly to the attrition-based thinking, which directed the organisational cultural thoughts towards composition that confirmed antagonism between gentlemen and lads . This setting has been absorbed and learned through cultural socialisation and was therefore not a product of the military organisation itself. The Finnish Armed Forces included two different communities (gentlemen and lads) within the same organisation as there were both the official and the unofficial organisations presented. This caused problems as they both made meaning-making processes simultaneously. These organisations had their own overlapping and in most cases also contradictory social meanings. The unofficial organisation has been overshadowed by the vast number of studies concerning the official organisation. The main reason for this systematic neglect is based on the reality of the attitudes and living conditions of the micro-level organisation which produced (perhaps) too realistic and repulsive viewpoints that are presenting a picture of a national level identity process in a way that is separating it from the ideals made to verify the ethos of national values. Complaining, griping, grumbling and moaning are usually situated in a category of abnormal and unwanted behaviour. However, within the context of a citizen soldier army community this was more of a characteristic feature of that organisation (in Finland) and therefore it was crucially important to locate the context of that abnormal behaviour. According to this study, it was not a malicious act but moreover seriously formed efforts in trying to use common sense in the chaos citizen soldiers faced when they were uniformed and placed in an unfamiliar process of disciplinary measures and frictions and competition between different ranks. There is much evidence that reinforces the argument that what seemed to be the most unconventional behaviour was finally the most efficient in a sense of military performance.

Structure-Function Studies of GDNF Family Ligand-RET signalling

Glial cell line-derived neurotrophic factor (GDNF) and its family members neurturin (NRTN), artemin (ARTN) and persephin (PSPN) are growth factors, which are involved in the development, differentiation and maintenance of many neuron types. In addition, they function outside of the nervous system, e.g. in the development of kidney, testis and liver. GDNF family ligand (GFL) signalling happens through a tetrameric receptor complex, which includes two glycosylphosphatidylinositol (GPI)-anchored GDNF family receptor (GFRα) molecules and two RET (rearranged during transfection) receptor tyrosine kinases. Each of the ligands binds preferentially one of the four GFRα receptors: GDNF binds to GFRα1, NRTN to GFRα2, ARTN to GFRα3 and PSPN to GFRα4. The signal is then delivered by RET, which cannot bind the GFLs on its own, but can bind the GFL-GFRα complex. Under normal cellular conditions, RET is only phosphorylated on the cell surface after ligand binding. At least the GDNF-GFRα1 complex is believed to recruit RET to lipid rafts, where downstream signalling occurs. In general, GFRαs consist of three cysteine-rich domains, but all GFRα4s except for chicken GFRα4 lack domain 1 (D1). We characterised the biochemical and cell biological properties of mouse PSPN receptor GFRα4 and showed that it has a significantly weaker capacity than GFRα1 to recruit RET to the lipid rafts. In spite of that, it can phosphorylate RET in the presence of PSPN and contribute to neuronal differentiation and survival. Therefore, the recruitment of RET to the lipid rafts does not seem to be crucial for the biological activity of all GFRα receptors. Secondly, we demonstrated that GFRα1 D1 stabilises the GDNF-GFRα1 complex and thus affects the phosphorylation of RET and contributes to the biological activity. This may be important in physiological conditions, where the concentration of the ligand or the soluble GFRα1 receptor is low. Our results also suggest a role for D1 in heparin binding and, consequently, in the biodistribution of released GFRα1 or in the formation of the GFL-GFRα-RET complex. We also presented the crystallographic structure of GDNF in the complex with GFRα1 domains 2 and 3. The structure differs from the previously published ARTN-GFRα3 structure in three significant ways. The biochemical data verify the structure and reveal residues participating in the interactions between GFRα1 and GDNF, and preliminarily also between GFRα1 and RET and heparin. Finally, we showed that, the precursor of the oncogenic MEN 2B (multiple endocrine neoplasia type 2) form of RET gets phosphorylated already during its synthesis in the endoplasmic reticulum (ER). We also demonstrated that it associates with Src homology 2 domain-containing protein (SHC) and growth factor receptor-bound protein (GRB2) in the ER, and has the capacity to activate several downstream signalling molecules.

Structure and function of GDNF receptor alpha splice variants

The growth factors of the glial cell line-derived neurotrophic factor (GDNF) family consisting of GDNF, neurturin (NRTN), artemin (ARTN) and persephin (PSPN), are involved in the development, differentiation and maintenance of many types of neurons. They also have important functions outside the nervous system in the development of kidney, testis and thyroid gland. Each of these GFLs preferentially binds to one of the glycosylphosphatidylinositol (GPI)-anchored GDNF family receptors α (GFRα). GDNF binds to GFRα1, NRTN to GFRα2, ARTN to GFRα3 and PSPN to GFRα4. The GFLs in the complex with their cognate GFRα receptors all bind to and signal through the receptor tyrosine kinase RET. Alternative splicing of the mouse GFRα4 gene yields three splice isoforms. These had been described as putative GPI-anchored, transmembrane and soluble forms. My goal was to characterise the function of the different forms of mouse GFRα4. I firstly found that the putative GPI-anchored GFRα4 (GFRα4-GPI) is glycosylated, membrane-bound, GPI-anchored and interacts with PSPN and RET. We also showed that mouse GFRα4-GPI mediates PSPN-induced phosphorylation of RET, promotes PSPN-dependent neuronal differentiation of the rat pheochromocytoma cell line PC6-3 and PSPN-dependent survival of cerebellar granule neurons (CGN). However, although this receptor can mediate PSPN-signalling and activate RET, GFRα4-GPI does not recruit RET into lipid rafts. The recruitment of RET into lipid rafts has previously been thought to be a crucial event for GDNF- and GFL-mediated signalling via RET. I secondly demonstrated that the putative transmembrane GFRα4 (GFRα4-TM) is indeed a real transmembrane GFRα4 protein. Although it has a weak binding capacity for PSPN, it can not mediate PSPN-dependent phosphorylation of RET, neuronal differentiation or survival. These data show that GFRα4-TM is inactive as a receptor for PSPN. Surprisingly, GFRα4-TM can negatively regulate PSPN-mediated signalling via GFRα4-GPI. GFRα4-TM interacts with GFRα4-GPI and blocks PSPN-induced phosphorylation of RET, neuronal differentiation as well as survival. Taken together, our data show that GFRα4-TM may act as a dominant negative inhibitor of PSPN-mediated signaling. The most exciting part of my work was the finding that the putative soluble GFRα4 (GFRα4-sol) can form homodimers and function as an agonist of the RET receptor. In the absence of PSPN, GFRα4-sol can promote the phosphorylation of RET, trigger the activation of the PI-3K/AKT pathway, induce neuronal differentiation and support the survival of CGN. Our findings are in line with a recent publication showing the GFRα4-sol might contribute to the inherited cancer syndrome multiple endocrine neoplasia type 2. Our data provide an explanation to how GFRα4-sol may cause or modify the disease. Mammalian GFRα4 receptors all lack the first Cys-rich domain which is present in other GFRα receptors. In the final part of my work I have studied the function of this particular domain. I created a truncated GFRα1 construct lacking the first Cys-rich domain. Using binding assays in both cellular and cell-free systems, phosphorylation assays with RET, as well as neurite outgrowth assays, we found that the first Cys-rich domain contributes to an optimal function of GFRα1, by stabilizing the interaction between GDNF and GFRα1.

Obstetric and gynaecological aspects of HIV infection

The purpose of the present study was to examine the outcome of pregnancies among HIV-infected women in Helsinki, use of the levonorgestrel-releasing intrauterine system (LNG-IUS) among HIV-infected women and the prevalence and risk factors of cytological and histologically proven cervical lesions in this population. Between 1993 and 2003 a total of 45 HIV-infected women delivered 52 singleton infants. HIV infection was diagnosed during pregnancy in 40% of the mothers. Seventeen of the mothers received antiretroviral (ARV) medication prior to pregnancy and in 34 cases, the medication was started during pregnancy. A good virological response (i.e. HIV RNA load <1000/mL during the last trimester) to ARV medication was achieved in 36/40 (90%) of the patients in whom HI viral load measurements were performed. Of the infants, 92% were born at term, and their mean (±SD) birth weight was 3350±395 g. The Caesarean section rate was low, 25%. All newborns received ARV medication and none of the infants born to mothers with pre-delivery diagnosis of maternal HIV infection were infected. The safety and advantages of the LNG-IUS were studied prospectively (n=12) and retrospectively (n=6). The LNG-IUS was well tolerated and no cases of PID or pregnancy were noted. Menstrual bleeding was reduced significantly during use of the LNG-IUS; this was associated with a slight increase in haemoglobin levels. Serum oestradiol concentrations remained in the follicular range in all subjects. The key finding was that genital shedding of HIV RNA did not change after the insertion of the LNG-IUS. The mean annual prevalence of low-grade squamous intraepithelial lesions (SIL) was 15% and that of high-grade SIL was 5% among 108 systematically followed HIV-infected women during 1989 2003. A reduced CD4 lymphocyte count was associated with an increased prevalence of SIL, whereas duration of HIV infection, use of ARV medication and HI viral load were not. The cumulative risk of any type of SIL was 17% after one year and 48% after five years among patients with initially normal Pap smears. The risk of developing SIL was associated with young age and a high initial HI viral load. During the follow-up 51 subjects (n=153) displayed cervical intraepithelial neoplasia (CIN), (16% CIN1 and 18% CIN 2-3). Only one case of cancer of the uterine cervix was detected. Pap smears were reliable in screening for CIN. Both nulliparity (p<0.01) and bacterial vaginosis (p<0.04) emerged as significant risk factors of CIN. In conclusion, a combination of universal antenatal screening and multidisciplinary management allows individualized treatment and prevents vertical transmission of HIV. Use of the LNG-IUS is safe among HIV-infected women and cervicovaginal shedding of HIV RNA is not affected by use of the LNG-IUS. The risk of cervical pre-malignant lesions is high among HIV-infected women despite systematic follow-up.