N-ethyl-N-nitrosourea (ENU) induced mutations within the Klotho gene lead to ectopic calcification and reduced lifespan in mouse models

Ectopic calcification (EC), which is the pathological deposition of calcium and phosphate in extra-skeletal tissues, may be associated with hypercalcaemic and hyperphosphataemic disorders, or it may occur in the absence of metabolic abnormalities. In addition, EC may be inherited as part of several monogenic disorders and studies of these have provided valuable insights into the metabolic pathways regulating mineral metabolism. For example, studies of tumoural calcinosis, a disorder characterised by hyperphosphataemia and progressive EC, have revealed mutations of fibroblast growth factor 23 (FGF23), polypeptide N-acetyl galactosaminyltransferase 3 (GALNT3) and klotho (KL), which are all part of a phosphate-regulating pathway. However, such studies in humans are limited by the lack of available large families with EC, and to facilitate such studies we assessed the progeny of mice treated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) for EC. This identified two mutants with autosomal recessive forms of EC, and reduced lifespan, designated Ecalc1 and Ecalc2. Genetic mapping localized the Ecalc1 and Ecalc2 loci to a 11.0 Mb region on chromosome 5 that contained the klotho gene (Kl), and DNA sequence analysis identified nonsense (Gln203Stop) and missense (Ile604Asn) Kl mutations in Ecalc1 and Ecalc2 mice, respectively. The Gln203Stop mutation, located in KL1 domain, was severely hypomorphic and led to a 17-fold reduction of renal Kl expression. The Ile604Asn mutation, located in KL2 domain, was predicted to impair klotho protein stability and in vitro expression studies in COS-7 cells revealed endoplasmic reticulum retention of the Ile604Asn mutant. Further phenotype studies undertaken in Ecalc1 (kl203X/203X) mice demonstrated elevations in plasma concentrations of phosphate, FGF23 and 1,25-dihydroxyvitamin D. Thus, two allelic variants of Kl that develop EC and represent mouse models for tumoural calcinosis have been established. © 2015 Esapa et al.

A novel LEMD3 mutation common to patients with osteopoikilosis with and without melorheostosis

Recent studies have reported loss of function mutations in the LEMD3 gene, encoding an inner nuclear membrane protein that influences Smad signaling, as a cause of osteopoikilosis, Buschke-Ollendorff syndrome, and melorheostosis. We investigated LEMD3 in a three-generation family with osteopoikilosis from the Azores, an affected father and daughter from Ireland with osteopoikilosis (the daughter also had melorheostosis), and two other individuals from the UK with isolated melorheostosis. We found a novel C to T substitution at position 2032 bp (cDNA) in exon 8 of LEMD3, resulting in a premature stop codon at amino acid position 678. This mutation co-segregates with the osteopoikilosis phenotype in both the Azorean family and the Irish family. It was not detected in any of the six unaffected family members or in 342 healthy Caucasian individuals. No LEMD3 mutations were detected in the two patients with sporadic melorheostosis. The LEMD3 mutation reported was clearly the cause of osteopoikilosis in the two families but its relationship to melorheostosis in one of the family members is still unclear. Perhaps unsurprisingly in what is a segmental disease, we did not find LEMD3 mutations in peripheral-blood-derived DNA from the two other individuals with sporadic melorheostosis. The nature of the additional genetic and/or environmental influences required for the development of melorheostosis in those with osteopoikilosis requires further investigation.

Genetic associations and functional characterization of M1 aminopeptidases and immune-mediated diseases

Endosplasmic reticulum aminopeptidase 1 (ERAP1), endoplasmic reticulum aminopeptidase 2 (ERAP2) and puromycin-sensitive aminopeptidase (NPEPPS) are key zinc metallopeptidases that belong to the oxytocinase subfamily of M1 aminopeptidase family. NPEPPS catalyzes the processing of proteosome-derived peptide repertoire followed by trimming of antigenic peptides by ERAP1 and ERAP2 for presentation on major histocompatibility complex (MHC) Class I molecules. A series of genome-wide association studies have demonstrated associations of these aminopeptidases with a range of immune-mediated diseases such as ankylosing spondylitis, psoriasis, Behçet's disease, inflammatory bowel disease and type I diabetes, and significantly, genetic interaction between some aminopeptidases and HLA Class I loci with which these diseases are strongly associated. In this review, we highlight the current state of understanding of the genetic associations of this class of genes, their functional role in disease, and potential as therapeutic targets.

The atmospheric extinction of light

An experiment is described that enables students to understand the properties of atmospheric extinction due to Rayleigh scattering. The experiment requires the use of red, green and blue lasers attached to a traveling microscope or similar device. The laser beams are passed through an artificial atmosphere, made from milky water, at varying depths, before impinging on either a light meter or a photodiode integral to a Picotech Dr. DAQ ADC. A plot of measured spectral intensity verses depth reveals the contribution Rayleigh scattering has to the extinction coefficient. For the experiment with the light meter, the extinction coefficient for red, green and blue light in the milky sample of water were 0.27, 0.36 and 0.47 cm-1 respectively and 0.032, 0.037 and 0.092 cm-1 for the Picotech Dr. DAQ ADC.

An account and analysis of the implementation of various ebook business models at QUT, Australia

QUT (Queensland University of Technology) is a leading university based in the city of Brisbane, Queensland, Australia and is a selectively research intensive university with 2,500 higher degree research students and an overall student population of 45,000 students. The transition from print to online resources is largely completed and the library now provides access to 450,000 print books, 1,000 print journals, 600,000 ebooks, 120,000 ejournals and 100,000 online videos. The ebook collection is now used three times as much as the print book collection. This paper focuses on QUT Library’s ebook strategy and the challenges of building and managing a rapidly growing collection of ebooks using a range of publishers, platforms, and business and financial models. The paper provides an account of QUT Library’s experiences in using Patron Driven Acquisition (PDA) using eBook Library (EBL); the strategic procurement of publisher and subject collections by lease and outright purchase models, the more recent transition to Evidence Based Selection (EBS) options provided by some publishers, and its piloting of etextbook models. The paper provides an in-depth analysis of each of these business models at QUT, focusing on access verses collection development, usage, cost per use, and value for money.

Text and Intensification of Its Impact in Chishti Samā`

In my master’s thesis I analyse mystical Islamic poetry in ritualistic performance context, samā` , focusing on the poetry used by the Chishti Sufis. The work is based on both literary sources and ethnographic material collected in India. The central textual source is Surūd-i Rūhānī, a compilation of mystical poetry. Textual sources, however, can be understood properly only in relation to the living performance context and therefore I also utilise interviews of Sufis and performers of mystical music and recordings of samā` assemblies along with texts. First part of the thesis concentrates on thematic overview of the poems and the process of selecting a suitable text for performance. The poems are written in three languages, viz. in Persian, Urdu and Hindi. Among the authors are both Sufis and non-Sufis. The poems, mystical and non-mystical alike, share the same poetic images and they acquire a mystical meaning when they are set to qawwali music and performed in samā` assemblies. My work includes several translations of verses not previously translated. Latter part of the thesis analyses the musical idiom of qawwali and the ways in which the impact of text on listeners is intensified in performance. Typically the intensification is accomplished in the level of a single poem through three different techniques: using introductory verses, inserting verses between the verses of the main poem and repeating individual units of text. The former two techniques are tied to creating a mystical state in the listeners while the latter aims at sustaining it. It is customary that a listener enraptured by mystical experience offers a monetary contribution to the performers. Thus, intensification of the text’s impact aims at enabling the listeners to experience mystical states.

Ivanov-mistik : Okkul'tnye motivy v poèzii i proze Vjačeslava Ivanova (1907-1919)

The subject of this work is the mysticism of Russian poet, critic and philosopher Vjacheslav Ivanov (1866-1949). The approach adopted involves the textual and discourse analysis and findings of the history of ideas. The subject has been considered important because of Ivanov's visions of his dead wife, writer Lydia Zinovieva-Annibal, which were combined with audible messages ("automatic writings"). Several automatic writings and descriptions of the visions from Ivanov's archive collections in St.Petersburg and Moscow are presented in this work. Right after the beginning of his hallucinations in the autumn of 1907, Ivanov was totally captivated by the theosophical ideas of Anna Mintslova, the background figure for this work. Anna Mintslova, a disciple of Rudolf Steiner's Esoteric School, offered Ivanov the theosophical concept of initiation to interpret paranormal phenomena in his intimate life. The work is divided into three main chapters, an introduction and aconclusion. The first chapter is called The Mystical Person: Anthropology of Ivanov and describes the role of the inner "Higher Self" in Ivanov's views on the nature of human consciousness. The political implications of the concepts, "mystical anarchism" and "sobornost" (religious unity) are also examined. The acquaintance and contacts with Anna Mintslova during 1906-1907 gave a framework to Ivanov's search for an organic society and personal religious experience. The second part, Mystics of Initiation and Visionary Aesthetics describes the influence of the initiation concept on Ivanov's aesthetic views (mainly "realistic symbolism"). On the other hand, some connections between the imagery of his visions and symbols in his verses of that period are established. Since Mintslova represented the ideas of Rudolf Steiner in Russia, several symbols shared by Steiner and Ivanov ("rose", "rose and cross") have been another subject of investigation. The preference for strict verse form in the lyrics of Ivanov's visionary period is interpreted as an attempt to place his own poetic creation within two traditions, a mystical and literary one. The third part of this work, Mystics of Hope and Terror, examines Ivanov's conception of Russia in connection with Mintslova's ideas of occult danger from the East. Ivanov's view of the "Russian idea" and his nationalistic idea during World War I are considered as a representation of the fear of the danger. Ivanov's interpretation of the October revolution is influenced by the theosophical concept of the "keeper of the threshold" which occurs in the context of the discourse of occult danger.

Konstrukcija mnimosti : K poètike "Voskovoj persony" Ju. Tynjanova

The subject of this work is the poetics of «The Wax Effigy», a short novel or novella by Jurii Tynianov, Russian writer, literary critic, historian of literature and prominent literary theoretician. The plot structure of the novel is based upon a real event, the creation by Bartolomeo Carlo Rastrelli in 1725 of a wax sculpture of the first Russian emperor, Peter the Great. «Construction of the Sham» consists of three chapters, an introduction and a conclusion. Due to the fact that Tynianov was at the same time a prose writer and theoretician of literature it seemed important to consider the reception of his prose and his works on literary theory in relationship to each other. The introduction is devoted to this task. The first chapter is about the history of the creation of the novel and its reception. Tynianov stopped writing one short story in order to write the novel; these two works have some common traits. It seems almost obvious that his work on the first text was a real step toward the creation of the second. In the first story there is an opposition of dead/alive which is semantic prefiguring of a central motif in «The Wax Effigy». An analysis of the reception of the novel demonstrated that almost every critic writing about the novel has described it as nonsense. Critics considered Tynianov's work in terms of «devices» and «content» and could not understand how devices are related to the content of the novel: the novel was thought as a signifier without any signified. Implicitly, critics thought the signified of the novel as a traditional one of the historical novel, as the historiosophical «idea», embodied in the system of literary devices. In this case literature becomes something instrumental, a kind of expression of extraliterary content. In contradistinction to that Tynianov considered literary semantics as an effect of the literary structure. From his point of view the literary sense is immanent to the process of signification accomplished inside the literary text. The second chapter is devoted to a rhetorical analysis of the opposition dead/alive. Tynianov systematically compares both terms of the opposition. As a result of this strategy the wax effigy of the dead emperor becomes «as if» alive and the world of living people «as if» dead. The qualifier «as if» refers to the fact that Tynianov creates an ambiguous semantic system. This rhetoric is related to European Romanticism and his «fantastic literature» (Merimé, Hoffmann, Maupassant etc.). But Tynianov demonstrates a linguistic origin of the strange fantoms created by romantics; he demystifies these idols by parodying the fantastic literature, that is, showing «how it was done». At the same time, the opposition mentioned above refers to his idea of «incongruity» which plays a prominent role in Tynianov s theory but has never been conceptualised. The incongruity is a inner collision of the literary text; from Tynianov's point of view the meaning of the work of literature is always a dynamic collision of semantically heterogeneous elements struggling with each other. In «The Wax Effigy» Tynianov creates a metalevel of the work demonstrating the process of creation of the literary sense. The third chapter is a reconstruction of Tynianov's conception of the historical prose, specifically of the mechanisms by which historical facts are transformed into literary events. Tynianov thought that the task of the historical novelist is to depict his hero as an actor, to demonstrate that as a wearer of many masks he is a creator of appearances, ambiguities. Here, in the «figure of fiction» (Andrei Belyi), the very idea of the historical prose and rhetoric employed in «The Wax Effigy», history and literature meet each other. In his last theoretical work, «On parody» Tynianov writes about the so-called sham structure of parody. In his opinion every parody is a text about other texts and «serious» work which could be read at the same time as a text about «reality». This twofold structure of parody is that of «The Wax Effigy»: that text speaks about ambiguities of the history and about ambiguities of the literary sense, about social reality of the past and - about the working of the literature itself. «The Wax Effigy» is written as a autoreflective text, as an experiment in literary semantics, as a system of literary ambiguities - of hero, rhetoric and the text itself. The meaning of the novel is created not by the embodiment extraliterary idea, but by the process of signification accomplished inside the work of literature. In this sense Tynianov's novel is parody, a break with the tradition of the historical novel preceding «The Wax Effigy».

Kääntäjän kyydissä uudelle vuosituhannelle : Miten Kornei Tšukovskin lastenrunot kotoutuvat suomalaiseen nykyaikaan?

Tutkielmassani tarkastelen venäläisen lastenkirjailijan ja käännösteoreetikon Kornei Tšukovskin (Kornej Čukovskij) riimisatuja ja runoja kääntäjän kannalta. Tavoitteenani on selvittää, miten noista lähes sata vuotta vanhoista teksteistä saisi suomennoksia, jotka täyttäisivät lastenkirjallisuudelle kulttuurissamme asetetut normit ja puhuttelisivat nykyistä lukijapolvea. Arvioin myös, olisivatko jo olemassa olevat, noin kolmekymmentä vuotta vanhat Tšukovski-suomennokset yhä riittävän ajanmukaisia julkaistaviksi uusintapainoksina. Tutkielman aineisto sisältää Tšukovskin tunnetuimmat, suurimmaksi osaksi 1920-luvulta peräisin olevat riimisadut ja runot sekä saduista tähän mennessä tehdyt suomennokset. Tutkielman teoreettisena taustana ovat manipulaatioteoria ja siitä edelleen kehitetty polysysteemiteoria. Kohdetekstipainotteisuutensa vuoksi ne muodostavat ihanteellisen viitekehyksen lastenkirjallisuuden kääntämiselle, jossa kohdekulttuurin sisäiset normit ovat aina voimakkaina läsnä. Sekä lähde- että kohdeteksteissä kiinnitän erityistä huomiota sellaisiin aineksiin, jotka vaatisivat ajanmukaistamista, mutta lisäksi paneudun lastenkirjallisuuden kääntämisen yleisiin kysymyksiin, kuten kotouttamiseen, ideologisiin ja pedagogisiin näkökohtiin, tekstin luettavuuteen, kuvan ja sanan suhteeseen sekä rytmiin ja riimiin. Suomennoksia arvioidessani pyrin ottamaan huomioon myös kirjailijan omat käännösihanteet. Käännösteoreetikkona Tšukovski piti kääntämisen tärkeimpänä lähtökohtana ehdotonta uskollisuutta lähdetekstin kirjoittajalle. Tutkielmani empiirisessä osassa käytän kahta eri tutkimusmenetelmää. Aluksi arvioin jo olemassa olevien käännösten ajanmukaisuutta. Sen jälkeen valitsen uusia tekstejä suomennettaviksi ja käännän niitä itse selvittääkseni, millaista manipulointia niiden nykyaikaistaminen edellyttäisi. Tutkimuksestani käy ilmi, että useimmat aiemmista Tšukovski-suomennoksista voisi julkaista vielä tänäkin päivänä. Niistä välittyy alkuperäisten tekstien nonsense-tyyli, ja ne täyttävät edelleen lastenkirjallisuutemme poeettiset normit. Ne pienet korjaukset, joita niihin saatettaisiin joutua tekemään, johtuvat lähinnä kohdekulttuurin arvoissa ja asenteissa tapahtuneista muutoksista. Esimerkiksi väkivalta ja rasismi ovat nykyään voimakkaampia tabuja kuin suomennosten ilmestymisen aikaan. Lisäksi Tšukovskilla on vielä suomentamattomia riimisatuja ja runoja, joista olisi mahdollista tehdä ajanmukaisia käännöksiä uusille lukijapolville. Tämä edellyttäisi kuitenkin tekstien manipulointia: alkuperäiset tekstit sisältävät aineksia, jotka ovat ristiriidassa lastenkirjallisuuden nykynormien kanssa. Osa riimisaduista vaatisi niin voimakasta manipulointia, että tuloksena olisi pikemminkin mukaelma kuin käännös. Koska tekstit ovat riimillisiä, jo pelkästään poeettiset näkökohdat aiheuttavat usein muutoksia semanttiseen sisältöön. Tšukovskin peräämä uskollisuus alkuperäiselle kirjailijalle tuntuukin jossakin määrin vanhentuneelta nykyisten käännösnormien aikana, jona kääntäminen katsotaan enemmän kulttuurienväliseksi viestinnäksi kuin lähde- ja kohdetekstien välisen samuuden etsimiseksi. Toivon tutkimukseni antavan hyödyllisiä viitteitä kustantajille, joka etsivät julkaistavakseen korkealuokkaista venäläistä lastenkirjallisuutta. Tšukovskin teokset kuuluvat maailman lastenkirjallisuuden klassikoihin, ja siksi myös nykyajan suomalaisille lukijoille kannattaisi tarjota mahdollisuutta tutustua niihin. Avainsanat – Nyckelord – Keywords Kornei Tšukovski, lastenkirjallisuuden kääntäminen, manipulaatio, polysysteeemi, normi, ideologia, rytmi, riimi Säilytyspaikka – Förvaringställe – Where deposited Humanistisen tiedekunnan kirjasto

Aavan meren täällä puolen : Arkkityyppiset piirteet ja amerikkalaisvaikutteet suosituimmissa suomalaisissa molli-iskelmissä

What are the musical features that turn a song into a hit? The aim of this research is to explore the musical features of hit tunes by studying the 224 most popular Finnish evergreens from the 1930s to the 1990s. It is remarkable, that 80-90% of Finnish oldies are in a minor key, though parallel major keys have also been widely employed within single pieces through, for example, modulations. Furthermore, melodies are usually diatonic, staying mostly in the same key. Consequently, chromatically altered tones in the melody and short modulations in the bridge sections become more prominent. I have concentrated in particular on the melodic lines in order to find the most typical melodic formulas from the data. These analyzed melodic formulas play an important role, because they serve as leading phrases and punchlines in songs. Analysis has revealed three major melodic formulas, which most often appear in the melodic lines of hit tunes. All of these formulas share common thematic ground, because they originate from the triadic tonic chord. Because the tonic chord is the most conventional opening chord in the verse parts, it is logical that these formulas occur most often in verses. The strong dominance of these formulas is very much a result of the rhythmic flexibility they possess; for instance, they can be found in every musical style from waltz to foxtrot. Alongside the major formulas lies a miscellaneous group of other tonic-related melodic formulas. One group of melodic formulas consists of melodic quotations. These quotations appear in a different musical context, for instance in a harmonically altered form, and are therefore often difficult to recognize as such. Yet despite the contextual manipulation, the distinctive character of the cited melody usually remains the same. Composers have also made use of certain popular chord-progressions in order to create new but familiar-sounding melodies. The most important individual progression in this case is what is known as a "circle of fifths" and its shortened, prolonged and altered versions. Because that progression is harmonically strong, it is also a contrastive tool used especially in chorus parts and middle sections (AABA). I have also paid attention to ragtime and jazz influences, which can be found in harmony parts and certain melody notes, which extend, suspend or alter the accompaning chords. Other influences from jazz and ragtime in the Finnish evergreen are evident in the use of typical Tin Pan Alley popular song forms. The most important is the AABA form, which dominates over the data along with the verse/chorus-type popular song form. To briefly illustrate the main results, the basic concept of the hit tune can be traced back to Tin Pan Alley songs, whereas the major stylistic aspects, such as minor keys and musical styles, bear influences from Russian, Western European, and Finnish traditions.

Auditory - Visual Matching in Learning Disabilities : Intervention Studies from Finland and Sweden

The present thesis discusses relevant issues in education: 1) learning disabilities including the role of comorbidity in LDs, and 2) the use of research-based interventions. This thesis consists of a series of four studies (three articles), which deepens the knowledge of the field of special education. Intervention studies (N=242) aimed to examine whether training using a nonverbal auditory-visual matching computer program had a remedial effect in different learning disabilities, such as developmental dyslexia, Attention Deficit Disorder (ADD) and Specific Language Impairment (SLI). These studies were conducted in both Finland and Sweden. The intervention’s non-verbal character made an international perspective possible. The results of the intervention studies confirmed, that the auditory-visual matching computer program, called Audilex had positive intervention effects. In Study I of children with developmental dyslexia there were also improvements in reading skills, specifically in reading nonsense words and reading speed. These improvements in tasks, which are thought to rely on phonological processing, suggest that such reading difficulties in dyslexia may stem in part from more basic perceptual difficulties, including those required to manage the visual and auditory components of the decoding task. In Study II the intervention had a positive effect on children with dyslexia; older students with dyslexia and surprisingly, students with ADD also benefited from this intervention. In conclusion, the role of comorbidity was apparent. An intervention effect was evident also in students’ school behavior. Study III showed that children with SLI experience difficulties very similar to those of children with dyslexia in auditory-visual matching. Children with language-based learning disabilities, such as dyslexia and SLI benefited from the auditory-visual matching intervention. Also comorbidity was evident among these children; in addition to formal diagnoses, comorbidity was explored with an assessment inventory, which was developed for this thesis. Interestingly, an overview of the data of this thesis shows positive intervention effects in all studies despite learning disability, language, gender or age. These findings have been described by a concept inter-modal transpose. Self-evidently these issues need further studies. In learning disabilities the aim in the future will also be to identify individuals at risk rather than by deficit; this aim can be achieved by using research-based interventions, intensified support in general education and inclusive special education. Keywords: learning disabilities, developmental dyslexia, attention deficit disorder, specific language impairment, language-based learning disabilities, comorbidity, auditory-visual matching, research-based interventions, inter-modal transpose

Using bacteria in banana for efficient use of fertilizer

This project aims to develop sustainable banana production practices by improving efficiency of fertilizer use. We will investigate how we can reintroduce endophytic beneficial bacteria that can be established inside the banana host to provide lasting and durable benefits to growth. In the current research we have been able to isolate bacteria from inside banana and preliminary characterisation indicates isolates with a range of attributes for improved efficiency of fertilizer uptake. Experimentation will include evaluation in vitro and pot trials and field trials using bacteria most likely to increase plant access to nutrients as well to compare nutrient impacts from conventional verses slow release fertilizer.

The Role Of Fumarase (FH) In Tumorigenesis

In this study, a predisposing gene for a recently characterized cancer syndrome, hereditary leiomyomatosis and renal cell cancer (HLRCC), was identified and the role of the gene was investigated in other familial cancers and in nonsyndromic tumorigenesis. HLRCC is a dominantly inherited disorder predisposing predominantly to uterine and skin leiomyomas, and also to renal cell cancer and uterine leiomyosarcoma. The disease gene was recently localized in Finnish families to 1q42-q43 by a genome-wide linkage search. Independently in the UK, a clinically similar condition, multiple cutaneous and uterine leiomyomata (MCUL), was linked to the same chromosomal region, strongly suggesting that HLRCC and MCUL are actually a single syndrome. Linkage results were confirmed by detecting loss of heterozygosity (LOH) at the disease locus in most of the patients' tumors, suggesting that this predisposing gene acts as a tumor suppressor. Through detailed investigation by genotyping of microsatellite markers and haplotype construction in Finnish and UK HLRCC/MCUL families we were able to narrow the disease locus down to 1.6 Mb. Extensive mutation screening of known and predicted transcripts in the target region resulted in identification of the HLRCC predisposing gene, fumarase (fumarate hydratase, FH). FH is a key enzyme in energy metabolism, catalyzing fumarate to malate in the tricarboxylic acid cycle (TCAC) in mitochondria. Germline alterations in FH segregating with the disease were detected in 25 of 42 HLRCC/MCUL families including whole-gene deletions, truncating small deletions/insertions and nonsense mutations, as well as substitutions or deletions of highly conserved amino acids. Biallelic inactivation was detected in almost all studied tumors of HLRCC patients. Furthermore, FH enzyme activity was reduced in the patients' normal tissues and was completely or virtually absent from tumors. Based on these findings, we extensively demonstrated that mutations in FH underlie the HLRCC/MCUL syndrome. In our studies of other familial cancers, evidence for involvement of FH defects was not found in familial prostate and breast cancers. To investigate the role of FH in sporadic tumorigenesis, we analyzed 652 lesions, including a series of 353 nonsyndromic counterparts of tumor types associated with HLRCC. Mutations in nonsyndromic tumors were rare and appeared to be limited to tumor types observed in the hereditary form of the disease. Biallelic inactivation of FH was detected in a uterine leiomyosarcoma, a cutaneous leiomyoma, a soft-tissue sarcoma, and in two uterine leiomyomas. In the uterine leiomyosarcoma and the cutaneous lesion FH mutations originated from the germline whereas the soft-tissue sarcoma harbored purely somatic changes. In uterine leiomyomas somatic mutations were detected in the two out of five tumors with LOH at the FH locus. Our findings demonstrate that FH inactivation is also involved in nonhereditary tumor development, and further support the hypothesis that FH acts as a tumor suppressor. The role of FH in predisposition to malignancies, renal cell carcinoma and leiomyosarcoma is important in the diagnosis and prevention of cancer among HLRCC patients. This study is of general clinical interest, because prior to our findings, little was known about the molecular genetics of uterine leiomyomas, the most common tumors of women.

The molecular basis of Marinesco-Sjögren syndrome

Marinesco-Sjögren syndrome (MSS) is a rare autosomal recessive neurodegenerative disorder characterized by cerebellar ataxia due to cerebellar cortical atrophy, infantile- or childhood-onset bilateral cataracts, progressive myopathy, and mild to severe mental retardation. Additional features include hypergonadotropic hypogonadism, various skeletal abnormalities, short stature, and strabismus. The neuroradiologic hallmarks are hypoplasia of both the vermis and cerebellar hemispheres. The histopathologic findings include severe cerebellar atrophy and loss of Purkinje and granule cells. The common pathologic findings in muscle biopsy are variation in muscle fiber size, atrophic fibers, fatty replacement, and rimmed vacuole formation. The presence of marked cerebellar atrophy with myopathy distinguishes MSS from another rare syndrome, the congenital cataracts, facial dysmorphism, and neuropathy syndrome (CCFDN). Previously, work by others had resulted in the identification of an MSS locus on chromosome 5q31. A subtype of MSS with myoglobinuria and neuropathy had been linked to the CCFDN locus on chromosome 18qter, at which mutations in the CTDP1 gene had been identified. We confirmed linkage to the previously identified locus on chromosome 5q31 in two Finnish families with eight affected individuals, reduced the critical region by fine-mapping, and identified SIL1 as a gene underlying MSS. We found a common homozygous founder mutation in all Finnish patients. The same mutation was also present in patient samples from Norway and Sweden. Altogether, we identified eight mutations in SIL1, including nonsense, frameshift, splice site alterations, and one missense mutation. SIL1 encodes a nucleotide exchange factor for the endoplasmic reticulum (ER) resident heat-shock protein 70 chaperone GRP78. GRP78 functions in protein synthesis and quality control of the newly synthesized polypeptides. It senses and responds to stressful cellular conditions. We showed that in mice, SIL1 and GRP78 show highly similar spatial and temporal tissue expression in developing and mature brain, eye, and muscle. Studying endogenous proteins in mouse primary hippocampal neurons, we found that SIL1 and GRP78 colocalize and that SIL1 localizes to the ER. We studied the subcellular localization of two mutant proteins, a missense mutant found in two patients and an artificial mutant lacking the ER retrieval signal, and found that both mutant proteins formed aggregates within the ER. Well in line with our findings and the clinical features of MSS, recent work by Zhao et al. showed that a truncation of SIL1 causes ataxia and cerebellar Purkinje cell loss in the naturally occurring woozy mutant mouse. Prior to Purkinje cell degeneration, the unfolded protein response is initiated and abnormal protein accumulations are present. MSS thus joins the group of protein misfolding and accumulation diseases. These findings highlight the importance of SIL1 and the role of the ER in neuronal function and survival. The results presented in this thesis provide tools for the molecular genetic diagnostics of MSS and give a basis for future studies on the molecular pathogenesis of MSS. Understanding the mechanisms behind this pleiotropic syndrome may provide insights into more common forms of ataxia, myopathy, and neurodegeneration.

Genetic basis of hereditary colorectal cancers: Hereditary nonpolyposis colorectal cancer and Familial adenomatous polyposis

Hereditary nonpolyposis colorectal cancer (HNPCC) and familial adenomatous polyposis (FAP) are characterized by a high risk and early onset of colorectal cancer (CRC). HNPCC is due to a germline mutation in one of the following MMR genes: MLH1, MSH2, MSH6 and PMS2. A majority of FAP and attenuated FAP (AFAP) cases are due to germline mutations of APC, causing the development of multiple colorectal polyps. To date, over 450 MMR gene mutations and over 800 APC mutations have been identified. Most of these mutations lead to a truncated protein, easily detected by conventional mutation detection methods. However, in about 30% of HNPCC and FAP, and about 90% of AFAP families, mutations remain unknown. We aimed to clarify the genetic basis and genotype-phenotype correlation of mutation negative HNPCC and FAP/AFAP families by advanced mutation detection methods designed to detect large genomic rearrangements, mRNA and protein expression alterations, promoter mutations, phenotype linked haplotypes, and tumoral loss of heterozygosity. We also aimed to estimate the frequency of HNPCC in Uruguayan CRC patients. Our expression based analysis of mutation negative HNPCC divided these families into two categories: 1) 42% of families linked to the MMR genes with a phenotype resembling that of mutation positive, and 2) 58% of families likely to be associated with other susceptibility genes. Unbalanced mRNA expression of MLH1 was observed in two families. Further studies revealed that a MLH1 nonsense mutation, R100X was associated with aberrant splicing of exons not related to the mutation and an MLH1 deletion (AGAA) at nucleotide 210 was associated with multiple exon skipping, without an overall increase in the frequency of splice events. APC mutation negative FAP/AFAP families were divided into four groups according to the genetic basis of their predisposition. Four (14%) families displayed a constitutional deletion of APC with profuse polyposis, early age of onset and frequent extracolonic manifestations. Aberrant mRNA expression of one allele was observed in seven (24%) families with later onset and less frequent extracolonic manifestations. In 15 (52%) families the involvement of APC could neither be confirmed nor excluded. In three (10%) of the families a germline mutation was detected in genes other than APC: AXIN2 in one family, and MYH in two families. The families with undefined genetic basis and especially those with AXIN2 or MYH mutations frequently displayed AFAP or atypical polyposis. Of the Uruguayan CRC patients, 2.6% (12/461) fulfilled the diagnostic criteria for HNPCC and 5.6% (26/461) were associated with increased risk of cancer. Unexpectedly low frequency of molecularly defined HNPCC cases may suggest a different genetic profile in the Uruguayan population and the involvement of novel susceptibility genes. Accurate genetic and clinical characterization of families with hereditary colorectal cancers, and the definition of the genetic basis of "mutation negative" families in particular, facilitate proper clinical management of such families.