Meän tapa puhua : Tornionlaakso pellolaisnuorten subjektiivisena paikkana ja murrealueena

This folk linguistic and human geographic study deals with dialect awareness, dialect use and place attachment. The study discusses theoretical and methodological issues current in sociolinguistics suggesting that the study of attitudes should be regarded as a core area in the study of variation and change. Furthermore, it is suggested that instead of putting effort into improving mental mapping methodology (adopted into folk linguistics from behavioural geography of the 1960 s), the more up-to-date thinking of space in geography should be adopted. The region and the dialect are treated as perceptual constructs in the study. The dialect perceptions of high school seniors in the Finnish Tornio Valley are examined trough a triangulation method involving a questionnaire, interviews and dialect recognition test as the research methods. The h in non-initial syllables (e.g. lähethä(ä)n, saunhaan ~ sauhnaan let s go into sauna ) turns out, expectedly, as the most salient feature in the dialect awareness of the locals and in terms of local identity construction. This feature is no longer heard in most of the present dialects of Finnish but is still thriving in the Tornio Valley in the cross-border dialect area. The metathetic variant (saunhaan > sauhnaan into sauna , käymhään > käyhmään to go ) is a characteristic feature of the Tornio Valley dialect. However, individual differences have long been found in the use of the h. This study challenges the essentialist variationist view of social categories (gender) by analysing variation from a quantitative but emic and human geographic point of view. The study shows that the variation of the h is statistically significantly patterned in terms of the degree of feeling of insideness vs. outsideness. New light is shed on the gender differences found in earlier sociolinguistic studies: differences in dialect use between and inside gender groups are illuminated by the fact that, in this case, it is young women who are generally less attached to the local community than young men, but this does not hold for all the individuals. The ideological motivation for preservation of the h seems to be based on the imagined community of Tornio Valley covering both the Swedish and the Finnish valley area. The general image of the dialect area and it s speakers, the shared cognitive dialect boundaries of the locals and the particularly deep level of awaress of the linguistic variation of the h are notable resources of the Tornio valley identity. Hyperdialectic forms analogical to the most frequently attested metathetic forms are found in the interview data, predicting that in this dialect the h will be maintained also in the future.

Kaunhiista kauhniiseen, satheesta satteeseen : Jälkitavuissa säilyneen h:n variaatiosta Pellossa vuoden 1995 aineiston valossa

Tutkielmassa tarkastellaan jälkitavuissa säilyneen, alun perin vokaalienvälisen h:n nykyedustusta Tornionlaakson Pellossa sosiolingvistisen teorian valossa. Keskeisenä tutkimusongelmana on variaation jasen muuttumisen suunnan selvittäminen, mitä tutkitaan sekä näennäis- että reaaliajallisen metodin turvin. Lisäksi pohditaan, millaiset kielensisäiset tekijät muutoksia ohjaavat. Reaaliaikaisen vertailun mahdollistavat Kirsi Kunnarin vuonna 1983 valmistuneen pro gradu -työn jälkitavujen h:ta koskevat tulokset Pellosta (informantit syntyneet vuosina 1898-1967). Pääpaino on kuitenkin näennäisajallisessa vertailussa; variaatiota tarkastellaan yksityiskohtaisesti erityisesti kielelliseltä kannalta, mikä on vaatinut perusteellista aineiston käsittelyä. Näennäisajallisessa tarkastelussa verrataankahden haastatellun ikäryhmän kieltä: nuorten (syntyneet vuosina 1986-1977) ja keski-ikäisten (syntyneet vuosina 1964-1945). Nuoria informantteja, joita on 14, on haastateltu pääosin pareittan. Vertailuryhmä, keski-ikäiset, koostuu heidän vanhem mistaan (7 informanttia). Nauhatunteja on yhteensä n. 13. Tulokset osoittavat, että toisin kuin muualla Peräpohjolassa, yleiskielen mukainen katovariantti (kou-luun) ei ole merkittävästi yleistynyt pellolaisten puhekielessä viimeisten neljän sukupolven aikana. Vokaalienvälisessä asemassa (koulu-h-un) h:ta ei nykypellolaisilla enää tapaa, ja ns. sisäheittoinenvariantti (koul-huun) on nuorten ryhmässä selvästi vähemmän suosiossa kuin keski-ikäisillä. Tornionjoki­laaksolainen, metateettinen h-variantti (kouh-luun) on yhä elinvoimainen: sen käyttö on nuorillakin lisääntynyt. Metateesi ei ole kuitenkaan kvantitatiivisesti juurikaan yleistynyt niissä fonotaktisissa ympäristöissä, joissa se aiheesta tehdyn väitöskirjan (1992) mukaan on ollut aiemmilla sukupolvilla harvinainen. Tätä saattaa osittain selittää tiettyjen puhekielisyyksien yleistyvä käyttö, mikä vaikuttaa eniten juuri näissä fonotaktisissa ympäristöissä siten, ettei metateesin toteutumiselle ole edellytyksiä. Pellon nykyisessä jälkitavujen h:n variaatiossa on osallisena vielä yksi variantti: assimiloitunut h (talhoon > talloon; toisheen > toisseen; kauphaan > kauppaan). Aiemmin tutkituilla sukupolvilla assimilaatiomuutos on ollut harvinainen, mutta nyt se on yleistynyt erityisesti nuorilla tytöillä. h:n assimilaatiossa syntyvät muodot lankeavat usein yhteen alueella jokseenkin fonemaattisena esiintyvän yleisgeminaation kanssa, osa muodoista muistuttaa erikoisgeminaatiota ja pieni osa yleiskielisiä muotoja. Tutkielmassa kyseenalaistetaan kielellisten seikkojen nojalla kyseisten muotojen tulkitseminen h:n assimilaatiosta johtuvaksi: assimiloituneiksi tulkituissa h-tapauksissa sekä konsonantin että sitä seuraavan vokaalin pituusdistribuutiot tukevat ajatusta, että ainakin yleisgeminaatiokonteksteissa assimilaatiomuodot ovat puhujan kannalta "mentaalista geminaatiota". Assimilaatiomuodot rinnastetaan tutkielmassa katomuotoihin sikäli, että niissäkin h on tasoittunut. Merkittävä ero on tavujen kvantiteettisuhteissa: assimilaatiomuodoissa ne säilyvät ennallaan (tal-hoon > tal-loon), katomuodoissa muuttuvat (ta-loon). h:n assimilaatiolla ja metateesilla epäillään aineistonperusteella olevan yhteisen tehtävän: sisäheittoisten talhoon, satheen -muotojen karttaminen tavujen kavantiteettisuhteita rikkomatta. Esitetyn hypoteesin mukaan syynä ovat suomen fonotaksille vieraat konsonantin + h:n yhtymät, jotka aikoinaan syntyivät murteeseen vokaalin heityttyä h:n edeltä.

HNbWO6 and HTaWO6: Novel oxides related to ReO3 formed by ion exchange of rutile-type LiNbWO6 and LiTaWO6

Both LiNbWO6 and LiTaWO6 undergo ion exchange in hot aqueous H2SO4 yielding the hydrates HMWO6 · H2O (M = Nb or Ta). The reaction is accompanied by a structural transformation from the rutile to the ReO3 structure. The cell constants are a = 3.783(3)Å for HNbWO6 · H2O and a = 3.785(5)Å for HTaWO6 · H2O. The ReO3 structure is retained by the dehydration products HMWO6 and MWO5.5 as well. HMWO6 phases yield H1+xMWO6 hydrogen bronzes on exposure to hydrogen in the presence of platinum catalyst.

Identtisten ja epäidenttisten kaksosten puhe : Monotsygoottisten ja ditsygoottisten kaksosten puheen vertailua kesto-, nopeus-, tauko- ja taajuusmittauksin

This thesis presents an experimental study of the speech prosody of identical and non-identical twins. Speech fluency, pauses, speech rate, utterance length and speech frequency were examined phonetically, auditorily, semantically and statistically. The methods included both reading tasks (reading the alphabet, numerical lists, sentences with foreign loan words, holiday theme questions as well as 1.5 pages of text with long sentences and complex words) and spontaneous speech tasks (picture description and answering holiday theme questions). The subjects were Finnish-speaking 22-28-year-old female twins: 8 identical (monozygotic) and 10 non-identical (dizygotic) pairs. One pair was male-female. Comparisons were made between twin groups and between sisters. The data was regathered from four twin pairs, to make it possible to investigate some subjects intra-individually. In addition phoneticians, phonetic students and people without knowledge of phonetic science were tested in two listening experiments. The results showed that the dizygotic twins differed more from each other than monozygotic twins and that monozygotic twin sisters shared more similarities than dizygotic twin sisters. For example, between monozygotic twin sisters smaller differences were found between word count, utterance length and speech rate in spontaneous speech tasks. Dizygotic twin sisters made more different kinds of reading mistakes with the same target words than monozygotic twin sisters, while monozygotic twin sisters made more of the same reading mistakes with the same target words than dizygotic twin sisters. The listening experiments showed that only professional phoneticians were able to recognize the twin sisters. Even though the twins had the possibility to freely choose their speech rate, pausing and speech frequency, they used their own speech patterns; these included the same average speech frequency, average speech rate, type of pausing routine or filled pauses, and other speech mannerisms throughout their speech.

Sosiaalisen verkostoanalyysin soveltuminen yhteisöllisen verkko-oppimisen tutkimiseen

Tietotekniikan kehittyessä lisääntyy sekä tietotyön tekijöiden tarve että paine tehostaa opetusta tietotekniikan avulla. Tietotyöläisten tarvitsemien taitojen opettamiseen tarvitaan uutta pedagogiikkaa, jossa korostetaan tietojen ja sääntöjen seuraamisen sijaan taitoja, itsenäisyyttä ja oppimaan oppimista. Tietotekniikankäyttäminen opetuksessa tuo omat haasteensa oppimistilanteisiin. Sosiokonstruktiivinen, yhteisöllinenverkko"-oppiminen onkin nousemassa perinteisen opettajajohtoisen opetuksen haastajaksi. Suomessa kehitetty Tutkiva oppiminen on yksi tämän viitekehyksen mukaisista pedagogisista menetelmistä. Koska sosiokonstruktiivinen teoria korostaa ihmisten välisiä suhteita yli yksittäisen ihmisen toiminnan, alan tutkimuksenkin on otettava kantaa tutkimustilanteessa esiintyviin vuorovaikutussuhteisiin. Tähän perinteiset psykologiset ja pedagogiset menetelmät eivät sovellu, kun taas sosiaalitieteissä suhteellisen vanha analyysimenetelmä, SVA eli sosiaalinen verkostoanalyysi, on tarkoitettu nimen\-omaan ihmisryhmien analysoimiseen. SVA:n sovellukset psykologian ja yhteisöllisen oppimisen alueilla ovat kuitenkin vasta alustavia, eikä luotettavaa näyttöä menetelmän soveltumisesta saatika hyödyllisistä tuloksista vielä ole. Tämän tutkimuksen on tarkoitus selvittää, onko SVA sovellettavissa tälle tutkimuksen alueelle. Tässä tutkimuksessa SVA:ta käytettiin 23 peruskoulu- ja lukioluokan Tutkivan oppimisen menetelmällä toteutettujen oppimistilanteiden tutkimiseen. Tulokset osoittavat, että SVA soveltuu yhteisöllisen verkko-oppimisen tutkimiseen, sillä tästä aineistosta löytyi alustavia SVA:n tunnuslukuja, jotka ovat yhteydessä oppimistilanteen laatuun. Tulokset ovat myös pitkälti sosiokonstruktiivisen teorian mukaisia: kurssin hyöty nousee, kun useat oppilaat kirjoittavat korkealaatuisia viestejä ja osallistuvat kattavasti eri keskusteluihin. Avainsanat: Tutkiva oppiminen, sosiokonstruktivismi, pedagogiikka, yhteisöllisyys, yhteisöllinen verkko-oppiminen, Fle3, SVA, sosiaalinen verkostoanalyysi, koulu, opetus.

"Sitte se menee koulus" : Eri kieli- ja kulttuuritaustaisen lapsen suomen kielen taito esiopetuksen päättövaiheessa

A child learns new things, creates social relationships and participates in play with the help of language. How can a child overcome these challenges if the surrounding language is not his mother tongue? The objective of learning a new language in the Pre-school education is an active bilingualism in all fields of the language. Theoretical context of the research rises from bilingualism, learning a language, language skills and evaluating them. Object of the research was to understand language skills of a child from a different linguistic and cultural background in the final stage of Pre-school education and to clarify how learning Finnish was supported during the Pre-school year. Answers to the research issues will be searched with the following questions: 1) What kind of language skills does a child from a different linguistic and cultural backgrounds have at the final stage of Pre-school education?, 1.1) What kind of listening comprehension skills?, 1.2) What kind of speech and vocabulary skills?, 1.3) What kind of structural skills?, 2) What kind of individual differences are there in language skills of children from different linguistic and cultural backgrounds?, and 3) How has a child from a different linguistic and cultural background been supported in learning Finnish during the Pre-school education? The view of language skills in this research is holistic even though it will be analysed in separate fields. The aim of this research is to form an overall impression of Finnish skills of the children participating in the research. Eight Pre-school-aged children with different linguistic and cultural backgrounds and their kindergarten teachers participated in this research. The children had taken part in Finnish activities for about three years. The research material consists of the test series (KITA), which evaluate children’s language skills – and of the questionnaire to the kindergarten teachers. The purpose of the questionnaire was to provide additional information on children’s language skills in Pre-school teaching situations and on supporting Finnish in Pre-school education. This research is qualitative and processing of the material is based on content analysis. According to the kindergarten teachers, the children’s social language skills were sufficient to cope in everyday life but children needed assistance with longer instructions. The same phenomenon could also be seen with the KITA tests – in which long and abstract instructions turned out to be difficult. Individual differences of the children were perceived in productivity skills, which were realised in fluent or influent speech. The children were supported in learning Finnish individually, in small-groups and in the activities of a whole group. ‘Finnish as the second language’ small-groups were the most common form of support in learning the language. The support at understanding activities was emphasized in whole group situations as well as in individual situations while assisting the child’s language skills. Generally, the children’s language skills were in the same level with developing basic language skills. The data of this research help to understand children’s language skills after three years of adopting Finnish. The results can be utilised in planning and evaluation of teaching another language.

Expression of functional GABAc receptors in the brain

γ-aminobutyric acid (GABA) is the main inhibitory transmitter in the nervous system and acts via three distinct receptor classes: A, B, and C. GABAC receptors are ionotropic receptors comprising ρ subunits. In this work, we aimed to elucidate the expression of ρ subunits in the postnatal brain, the characteristics of ρ2 homo-oligomeric receptors, and the function of GABAC receptors in the hippocampus. In situ hybridization on rat brain slices showed ρ2 mRNA expression from the newborn in the superficial grey layer of the superior colliculus, from the first postnatal week in the hippocampal CA1 region and the pretectal nucleus of the optic tract, and in the adult dorsal lateral geniculate nucleus. Quantitative RT-PCR revealed expression of all three ρ subunits in the hippocampus and superior colliculus from the first postnatal day. In the hippocampus, ρ2 mRNA expression clearly dominated over ρ1 and ρ3. GABAC receptor protein expression was confirmed in the adult hippocampus, superior colliculus, and dorsal lateral geniculate nucleus by immunohistochemistry. From the selective distribution of ρ subunits, GABAC receptors may be hypothesized to be specifically involved in aspects of visual image motion processing in the rat brain. Although previous data had indicated a much higher expression level for ρ2 subunit transcripts than for ρ1 or ρ3 in the brain, previous work done on Xenopus oocytes had suggested that rat ρ2 subunits do not form functional homo-oligomeric GABAC receptors but need ρ1 or ρ3 subunits to form hetero-oligomers. Our results demonstrated, for the first time, that HEK 293 cells transfected with ρ2 cDNA displayed currents in whole-cell patch-clamp recordings. Homomeric rat ρ2 receptors had a decreased sensitivity to, but a high affinity for picrotoxin and a marked sensitivity to the GABAC receptor agonist CACA. Our results suggest that ρ2 subunits may contribute to brain function, also in areas not expressing other ρ subunits. Using extracellular electrophysiological recordings, we aimed to study the effects of the GABAC receptor agonists and antagonists on responses of the hippocampal neurons to electrical stimulation. Activation of GABAC receptors with CACA suppressed postsynaptic excitability and the GABAC receptor antagonist TPMPA inhibited the effects of CACA. Next, we aimed to display the activation of the GABAC receptors by synaptically released GABA using intracellular recordings. GABA-mediated long-lasting depolarizing responses evoked by high-frequency stimulation were prolonged by TPMPA. For weaker stimulation, the effect of TPMPA was enhanced after GABA uptake was inhibited. Our data demonstrate that GABAC receptors can be activated by endogenous synaptic transmitter release following strong stimulation or under conditions of reduced GABA uptake. The lack of GABAC receptor activation by less intensive stimulation under control conditions suggests that these receptors are extrasynaptic and activated via spillover of synaptically released GABA. Taken together with the restricted expression pattern of GABAC receptors in the brain and their distinctive pharmacological and biophysical properties, our findings supporting extrasynaptic localization of these receptors raise interesting possibilities for novel pharmacological therapies in the treatment of, for example, epilepsy and sleep disorders.

Xanthine oxidoreductase in essential hypertension and metabolic syndrome : Experimental studies on rodent models

Hypertension, obesity, dyslipidemia and dysglycemia constitute metabolic syndrome, a major public health concern, which is associated with cardiovascular mortality. High dietary salt (NaCl) is the most important dietary risk factor for elevated blood pressure. The kidney has a major role in salt-sensitive hypertension and is vulnerable to harmful effects of increased blood pressure. Elevated serum urate is a common finding in these disorders. While dysregulation of urate excretion is associated with cardiovascular diseases, present studies aimed to clarify the role of xanthine oxidoreductase (XOR), i.e. xanthine dehydrogenase (XDH) and its post-translational isoform xanthine oxidase (XO), in cardiovascular diseases. XOR yields urate from hypoxanthine and xanthine. Low oxygen levels upregulate XOR in addition to other factors. In present studies higher renal XOR activity was found in hypertension-prone rats than in the controls. Furthermore, NaCl intake increased renal XOR dose-dependently. To clarify whether XOR has any causal role in hypertension, rats were kept on NaCl diets for different periods of time, with or without a XOR inhibitor, allopurinol. While allopurinol did not alleviate hypertension, it prevented left ventricular and renal hypertrophy. Nitric oxide synthases (NOS) produce nitric oxide (NO), which mediates vasodilatation. A paucity of NO, produced by NOS inhibition, aggravated hypertension and induced renal XOR, whereas NO generating drug, alleviated salt-induced hypertension without changes in renal XOR. Zucker fa/fa rat is an animal model of metabolic syndrome. These rats developed substantial obesity and modest hypertension and showed increased hepatic and renal XOR activities. XOR was modified by diet and antihypertensive treatment. Cyclosporine (CsA) is a fungal peptide and one of the first-line immunosuppressive drugs used in the management of organ transplantation. Nephrotoxicity ensue high doses resulting in hypertension and limit CsA use. CsA increased renal XO substantially in salt-sensitive rats on a high NaCl diet, indicating a possible role for this reactive oxygen species generating isoform in CsA nephrotoxicity. Renal hypoxia, common to these rodent models of hypertension and obesity, is one of the plausible XOR inducing factors. Although XOR inhibition did not prevent hypertension, present experimental data indicate that XOR plays a role in the pathology of salt-induced cardiac and renal hypertrophy.

Androgen receptor in transcriptional regulation and carcinogenesis

Androgens control a variety of developmental processes that create the male phenotype and are important for maintaining male fertility and normal functions of tissues and organs that are not directly involved in procreation. Androgen receptor (AR) that mediates the biological actions of androgens is a member of the nuclear receptor superfamily of ligand-inducible transcription factors. Although AR was cloned over 15 years ago, the mechanisms by which it regulates gene expression are not well understood. A growing body of in vitro experimental evidence suggests that a complex network of proteins is involved in the androgen-dependent transcriptional regulation. However, the process of AR-dependent transcriptional regulation under physiological conditions is largely elusive. In the present study, a series of experiments were performed, including quantitative chromatin immunoprecipitation (ChIP) assays, to investigate AR-mediated transcription process using living prostate cancer cells. Our results show that the loading of AR and recruitment of coactivators and RNA polymerase II (Pol II) to both the promoter and enhancer of AR target genes are a transient and cyclic event that in addition to hyperacetylation, also involves dynamic changes in methylation, phosphorylation of core histone H3 in androgen-treated LNCaP cells. The dynamics of testosterone (T)-induced loading of AR onto the proximal promoters of the genes clearly differed from that loaded onto the distal enhancers. Significantly, more holo-AR was loaded onto the enhancers than the promoters, but the principal Pol II transcription complex was assembled on the promoters. By contrast, the pure antiandrogen bicalutamide (CDX) complexed to AR elicited occupancy of the PSA promoter, but was unable to load onto the PSA enhancer and was incapable of recruiting Pol II, coactivators and following changes of covalent histone modifications. The partial antagonist cyproterone acetate (CPA) and mifepristone (RU486) were capable of promoting AR loading onto both the PSA promoter and enhancer at a comparable efficiency with androgen in LNCaP cells expressing mutant AR. However, CPA- and RU486-bound AR not only recruited Pol II and coactivator p300 and GRIP1 onto the promoter and enhancer, but also recruited the corepressor NCoR onto the promoter as efficiently as CDX. In addition, we demonstrate that both proteasome and protein kinases are implicated in AR-mediated transcription. Even though proteasome inhibitor MG132 and protein kinase inhibitor DRB (5, 6-Dichlorobenzimidazole riboside) can block ligand-dependent accumulation of PSA mRNA with same efficiency, their use results in different molecular profiles in terms of the formation of AR-mediated transcriptional complex. Collectively, these results indicate that transcriptional activation by AR is a complicated process, which includes transient loading of holo-AR and recruitment of Pol II and coregulators accompanied by a cascade of distinct covalent histone modifications; This process involves both the promoter and enhancer elements, as well as other general components of the cell machineries e.g. proteasome and protein kinase; The pure antiandrogen CDX and the partial antagonist CPA and RU486 exhibit clearly different profiles in terms of their ability to induce the formation of AR-dependent transcriptional complexes and the histone modifications associated with the target genes in human prostate cancer cells. Finally, by using quantitative RT-PCR to compare the expression of sixteen AR co-regulators in prostate cancer cell lines, xenografts, and clinical prostate cancer specimens we suggest that AR co-regulators protein inhibitor of activated STAT1 (PIAS1) and steroid receptor coactivator 1(SRC1) could be involved in the progression of prostate cancer.

Monolithic surface acoustic wave resonator oscillator

A monolithic surface acoustic wave (SAW) resonator operating at 156 MHz, in which the frequency controlling element is a Fabry–Perot type of SAW resonator and the gain element is a monolithic SAW amplifier (SiOx/InSb/SiOx structure located inside the SAW resonator cavity) is described and experimental details presented. Based on the existing experimental data, an uhf monolithic ring resonator oscillator is proposed. Journal of Applied Physics is copyrighted by The American Institute of Physics.

Microstructural study of rapidly solidified titanium eutectoid alloysstar, open

Rapid solidification of Ti-7.3wt.%Cu (near-eutectoid composition), Ti-36.2wt.%Ni and Ti-34.3wt.% Ni-5.8wt.%Si alloys has been carried out by electron beam melting and splat quenching on a water-cooled rotating copper disc. The product obtained was in the form of thin ribbons 60–100 μm thick. Transmission electron microscopy studies of Ti---Cu alloy splats showed that the microstructure consisted of a mixture of martensite and a lamellar eutectoid product. The formation of the intermetallic compound Ti2Cu involved a diffusionless ω transformation and spinodal clustering. In the case of Ti---Ni alloy the as-quenched microstructure is complex, consisting of α, transformed β and intermetallic phases. This could have arisen possibly as a result of local variation in cooling rates. Rapid solidification of Ti---Ni---Si alloy resulted in partial quenching of an amorphous phase. The amorphous phase was seen to be extremely hard (a Vickers hardness of about 800 HV).

Molecular Mechanisms of Androgen Receptor Interactions

The androgen receptor (AR) mediates the effects of the male sex-steroid hormones (androgens), testosterone and 5?-dihydrotestosterone. Androgens are critical in the development and maintenance of male sexual characteristics. AR is a member of the steroid receptor ligand-inducible transcription factor family. The steroid receptor family is a subgroup of the nuclear receptor superfamily that also includes receptors for the active forms of vitamin A, vitamin D3, and thyroid hormones. Like all nuclear receptors, AR has a conserved modular structure consisting of a non-conserved amino-terminal domain (NTD), containing the intrinsic activation function 1, a highly conserved DNA-binding domain, and a conserved ligand-binding domain (LBD) that harbors the activation function 2. Each of these domains plays an important role in receptor function and signaling, either via intra- and inter-receptor interactions, interactions with specific DNA sequences, termed hormone response elements, or via functional interactions with domain-specific proteins, termed coregulators (coactivators and corepressors). Upon binding androgens, AR acquires a new conformational state, translocates to the nucleus, binds to androgen response elements, homodimerizes and recruits sequence-specific coregulatory factors and the basal transcription machinery. This set of events is required to activate gene transcription (expression). Gene transcription is a strictly modulated process that governs cell growth, cell homeostasis, cell function and cell death. Disruptions of AR transcriptional activity caused by receptor mutations and/or altered coregulator interactions are linked to a wide spectrum of androgen insensitivity syndromes, and to the pathogenesis of prostate cancer (CaP). The treatment of CaP usually involves androgen depletion therapy (ADT). ADT achieves significant clinical responses during the early stages of the disease. However, under the selective pressure of androgen withdrawal, androgen-dependent CaP can progress to an androgen-independent CaP. Androgen-independent CaP is invariably a more aggressive and untreatable form of the disease. Advancing our understanding of the molecular mechanisms behind the switch in androgen-dependency would improve our success of treating CaP and other AR related illnesses. This study evaluates how clinically identified AR mutations affect the receptor s transcriptional activity. We reveal that a potential molecular abnormality in androgen insensitivity syndrome and CaP patients is caused by disruptions of the important intra-receptor NTD/LBD interaction. We demonstrate that the same AR LBD mutations can also disrupt the recruitment of the p160 coactivator protein GRIP1. Our investigations reveal that 30% of patients with advanced, untreated local CaP have somatic mutations that may lead to increases in AR activity. We report that somatic mutations that activate AR may lead to early relapse in ADT. Our results demonstrate that the types of ADT a CaP patient receives may cause a clustering of mutations to a particular region of the receptor. Furthermore, the mutations that arise before and during ADT do not always result in a receptor that is more active, indicating that coregulator interactions play a pivotal role in the progression of androgen-independent CaP. To improve CaP therapy, it is necessary to identify critical coregulators of AR. We screened a HeLa cell cDNA library and identified small carboxyl-terminal domain phosphatase 2 (SCP2). SCP2 is a protein phosphatase that directly interacts with the AR NTD and represses AR activity. We demonstrated that reducing the endogenous cellular levels of SCP2 causes more AR to load on to the prostate specific antigen (PSA) gene promoter and enhancer regions. Additionally, under the same conditions, more RNA polymerase II was recruited to the PSA promoter region and overall there was an increase in androgen-dependent transcription of the PSA gene, revealing that SCP2 could play a role in the pathogenesis of CaP.

Studies on the pro-oxidative properties and neurotoxic potential of Angeli's salt-derived nitroxyl (HNO/NO-)

The biological function of nitric oxide and its oxidized forms has received a great deal of attention over the past two decades. However much less attention has been focused on the reduced nitric oxide, nitroxyl (HNO). Unlike NO, HNO is highly reactive species and thus it needs to be generated by using donor compounds under experimental conditions. Currently there is only one donor available, Angeli s salt, which releases HNO in a controlled fashion under pysiological conditions. Prior studies have shown the pro-oxidative and cytotoxic potential of Angeli s salt compared to NO donors. The high reactivity of HNO with cysteine thiols is considered to form the biochemical basis for its unique properties compared to other nitrogen oxides. Such thiol modification cold result in disturbances of vital cellular functions and subsequently to death of disturbance sensitive cells, such as neurons. Therefore modification of proteins and lipids was studied in vitro and the potential neurotoxicity was studied in vivo by local infusion of Angeli s salt into the rat central nervous system. The results show that under aerobic in vitro conditions, HNO can, subsequent to autoxidation, cause irreversible oxidative modification of proteins and lipids. These effects are not however seen in cell culture or following infusion of Angeli s salt directly into the rat central nervous tissue likely due to presence of lower oxygen and higher thiol concentration. However, due to high reactivity with thiols, HNO can cause irreversible inactivation of cysteine modification sensitive enzymes such as cysteine proteases papain in vitro and cathepsin B in cell culture. Furthermore it was shown that infusion of HNO releasing Angeli s salt into the rat central nervous system causes necrotic cell death and motor dysfunction following infusion into the lumbal intrathecal space. In conclusion, the acute neurotoxic potential of Angeli s salt was shown to be relatively low, but still higher compared to NO donors. HNO was shown to affect numerous cellular processes which could result in neurotoxicity if HNO was produced in vivo.

Role of Basement Membrane and Extracellular Matrix Proteins in the Adhesion and Spreading of Immortalized Human Corneal Epithelial Cells

The repair of corneal wounds requires both epithelial cell adhesion and migration. Basement membrane (BM) and extracellular matrix (ECM) proteins function in these processes via integrin and non-integrin receptors. We have studied the adhesion, spreading and migration of immortalized human corneal epithelial (HCE) cells and their interactions with the laminins (Lms), fibronectins and tenascins produced. Human corneal BM expresses Lms-332 and -511, while Lm-111 was not found in these experiments. HCE cells produced both processed and unprocessed Lm-332, whereas neither Lm-111 nor Lm-511 was produced. Because HCE cells did not produce Lm-511, although it was present in corneal BM, we suggest that Lm-511 is produced by stromal keratocytes. The adhesion of HCE cells to Lms-111, -332 and -511 was studied first by determining the receptor composition of HCE cells and then by using quantitative cell adhesion assays. Immunofluorescence studies revealed the presence of integrin α2, α3, α6, β1 and β4 subunits. Among the non-integrin receptors, Lutheran (Lu) was found on adhering HCE cells. The cells adhered via integrin α3β1 to both purified human Lms-332 and -511 as well as to endogenous Lm-332. However, only integrin β1 subunit functioned in HCE cell adhesion to mouse Lm-111. The adhesion of HCE cells to Lm-511 was also mediated by Lu. Since Lm-511 did not induce Lu into focal adhesions in HCE cells, we suggest that Lm-511 serves as an ECM ligand enabling cell motility. HCE cells produced extradomain-A fibronectin, oncofetal fibronectin and tenascin-C (Tn-C), which are also found during corneal wound healing. Monoclonal antibodies (MAbs) against integrins α5β1 and αvβ6 as well as the arginine-glycine-aspartic acid (RGD) peptide inhibited the adhesion of HCE cells to fibronectin. Although the cells did not adhere to Tn-C, they adhered to the fibronectin/Tn-C coat and were then more efficiently inhibited by the function-blocking MAbs and RGD peptide. During the early adhesion, HCE cells codeposited Lm-332 and the large subunit of tenascin-C (Tn-CL) beneath the cells via the Golgi apparatus and microtubules. Integrin β4 subunit, which is a hemidesmosomal component, did not mediate the early adhesion of HCE cells to Lm-332 or Lm-332/Tn-C. Based on these results, we suggest that the adhesion of HCE cells is initiated by Lm-332 and modulated by Tn-CL, as it has been reported to prevent the assembly of hemidesmosomes. Thereby, Tn-CL functions in the motility of HCE cells during wound healing. The different distribution of processed and unprocessed Lm-332 in adhering, spreading and migrating HCE cells suggests a distinct role for these isoforms. We conclude that the processed Lm-332 functions in cell adhesion, whereas the unprocessed Lm-332 participates in cell spreading and migration.

Familial aggregation of type 1 diabetes and diabetic nephropathy in Finland

Type 1 diabetes (T1D) is a common, multifactorial disease with strong familial clustering. In Finland, the incidence of T1D among children aged 14 years or under is the highest in the world. The increase in incidence has been approximately 2.4% per year. Although most new T1D cases are sporadic the first-degree relatives are at an increased risk of developing the same disease. This study was designed to examine the familial aggregation of T1D and one of its serious complications, diabetic nephropathy (DN). More specifically the study aimed (1) to determine the concordance rates of T1D in monozygotic (MZ) and dizygotic (DZ) twins and to estimate the relative contributions of genetic and environmental factors to the variability in liability to T1D as well as to study the age at onset of diabetes in twins; (2) to obtain long-term empirical estimates of the risk of T1D among siblings of T1D patients and the factors related to this risk, especially the effect of age at onset of diabetes in the proband and the birth cohort effect; (3) to establish if DN is aggregating in a Finnish population-based cohort of families with multiple cases of T1D, and to assess its magnitude and particularly to find out whether the risk of DN in siblings is varying according to the severity of DN in the proband and/or the age at onset of T1D: (4) to assess the recurrence risk of T1D in the offspring of a Finnish population-based cohort of patients with childhood onset T1D, and to investigate potential sex-related effects in the transmission of T1D from the diabetic parents to their offspring as well as to study whether there is a temporal trend in the incidence. The study population comprised of the Finnish Young Twin Cohort (22,650 twin pairs), a population-based cohort of patients with T1D diagnosed at the age of 17 years or earlier between 1965 and 1979 (n=5,144) and all their siblings (n=10,168) and offspring (n=5,291). A polygenic, multifactorial liability model was fitted to the twin data. Kaplan-Meier analyses were used to provide the cumulative incidence for the development of T1D and DN. Cox s proportional hazards models were fitted to the data. Poisson regression analysis was used to evaluate temporal trends in incidence. Standardized incidence ratios (SIRs) between the first-degree relatives of T1D patients and background population were determined. The twin study showed that the vast majority of affected MZ twin pairs remained discordant. Pairwise concordance for T1D was 27.3% in MZ and 3.8% in DZ twins. The probandwise concordance estimates were 42.9% and 7.4%, respectively. The model with additive genetic and individual environmental effects was the best-fitting liability model to T1D, with 88% of the phenotypic variance due to genetic factors. The second paper showed that the 50-year cumulative incidence of T1D in the siblings of diabetic probands was 6.9%. A young age at diagnosis in the probands considerably increased the risk. If the proband was diagnosed at the age of 0-4, 5-9, 10-14, 15 or more, the corresponding 40-year cumulative risks were 13.2%, 7.8%, 4.7% and 3.4%. The cumulative incidence increased with increasing birth year. However, SIR among children aged 14 years or under was approximately 12 throughout the follow-up. The third paper showed that diabetic siblings of the probands with nephropathy had a 2.3 times higher risk of DN compared with siblings of probands free of nephropathy. The presence of end stage renal disease (ESRD) in the proband increases the risk three-fold for diabetic siblings. Being diagnosed with diabetes during puberty (10-14) or a few years before (5-9) increased the susceptibility for DN in the siblings. The fourth paper revealed that of the offspring of male probands, 7.8% were affected by the age of 20 compared with 5.3% of the offspring of female probands. Offspring of fathers with T1D have 1.7 times greater risk to be affected with T1D than the offspring of mothers with T1D. The excess risk in the offspring of male fathers manifested itself through the higher risk the younger the father was when diagnosed with T1D. Young age at onset of diabetes in fathers increased the risk of T1D greatly in the offspring, but no such pattern was seen in the offspring of diabetic mothers. The SIR among offspring aged 14 years or under remained fairly constant throughout the follow-up, approximately 10. The present study has provided new knowledge on T1D recurrence risk in the first-degree relatives and the risk factors modifying the risk. Twin data demonstrated high genetic liability for T1D and increased heritability. The vast majority of affected MZ twin pairs, however, remain discordant for T1D. This study confirmed the drastic impact of the young age at onset of diabetes in the probands on the increased risk of T1D in the first-degree relatives. The only exception was the absence of this pattern in the offspring of T1D mothers. Both the sibling and the offspring recurrence risk studies revealed dynamic changes in the cumulative incidence of T1D in the first-degree relatives. SIRs among the first-degree relatives of T1D patients seems to remain fairly constant. The study demonstrates that the penetrance of the susceptibility genes for T1D may be low, although strongly influenced by the environmental factors. Presence of familial aggregation of DN was confirmed for the first time in a population-based study. Although the majority of the sibling pairs with T1D were discordant for DN, its presence in one sibling doubles and presence of ESRD triples the risk of DN in the other diabetic sibling. An encouraging observation was that although the proportion of children to be diagnosed with T1D at the age of 4 or under is increasing, they seem to have a decreased risk of DN or at least delayed onset.