The Alzheimer`s Association external quality control program for cerebrospinal fluid biomarkers

Background: The cerebrospinal fluid (CSF) biomarkers amyloid beta (A beta)-42, total-tau (T-tau), and phosphorylated-tau (P-tau) demonstrate good diagnostic accuracy for Alzheimer`s disease (AD). However, there are large variations in biomarker measurements between studies, and between and within laboratories. The Alzheimer`s Association has initiated a global quality control program to estimate and monitor variability of measurements, quantify batch-to-batch assay variations, and identify sources of variability. In this article, we present the results from the first two rounds of the program. Methods: The program is open for laboratories using commercially available kits for A beta, T-tau, or P-tau. CSF samples (aliquots of pooled CSF) are sent for analysis several times a year from the Clinical Neurochemistry Laboratory at the Molndal campus of the University of Gothenburg, Sweden. Each round consists of three quality control samples. Results: Forty laboratories participated. Twenty-six used INNOTEST enzyme-linked immunosorbent assay kits, 14 used Luminex xMAP with the INNO-BIA AlzBio3 kit (both measure A beta-(1-42), P-tau(181P), and T-tau), and 5 used Mesa Scale Discovery with the A beta triplex (A beta N-42, A beta N-40, and A beta N-38) or T-tau kits. The total coefficients of variation between the laboratories were 13% to 36%. Five laboratories analyzed the samples six times on different occasions. Within-laboratory precisions differed considerably between biomarkers within individual laboratories. Conclusions: Measurements of CSF AD biomarkers show large between-laboratory variability, likely caused by factors related to analytical procedures and the analytical kits. Standardization of laboratory procedures and efforts by kit vendors to increase kit performance might lower variability, and will likely increase the usefulness of CSF AD biomarkers. (C) 2011 The Alzheimer`s Association. All rights reserved.

Rare genomic structural variants in complex disease: lessons from the replication of associations with obesity.

The limited ability of common variants to account for the genetic contribution to complex disease has prompted searches for rare variants of large effect, to partly explain the 'missing heritability'. Analyses of genome-wide genotyping data have identified genomic structural variants (GSVs) as a source of such rare causal variants. Recent studies have reported multiple GSV loci associated with risk of obesity. We attempted to replicate these associations by similar analysis of two familial-obesity case-control cohorts and a population cohort, and detected GSVs at 11 out of 18 loci, at frequencies similar to those previously reported. Based on their reported frequencies and effect sizes (OR≥25), we had sufficient statistical power to detect the large majority (80%) of genuine associations at these loci. However, only one obesity association was replicated. Deletion of a 220 kb region on chromosome 16p11.2 has a carrier population frequency of 2×10(-4) (95% confidence interval [9.6×10(-5)-3.1×10(-4)]); accounts overall for 0.5% [0.19%-0.82%] of severe childhood obesity cases (P = 3.8×10(-10); odds ratio = 25.0 [9.9-60.6]); and results in a mean body mass index (BMI) increase of 5.8 kg.m(-2) [1.8-10.3] in adults from the general population. We also attempted replication using BMI as a quantitative trait in our population cohort; associations with BMI at or near nominal significance were detected at two further loci near KIF2B and within FOXP2, but these did not survive correction for multiple testing. These findings emphasise several issues of importance when conducting rare GSV association, including the need for careful cohort selection and replication strategy, accurate GSV identification, and appropriate correction for multiple testing and/or control of false discovery rate. Moreover, they highlight the potential difficulty in replicating rare CNV associations across different populations. Nevertheless, we show that such studies are potentially valuable for the identification of variants making an appreciable contribution to complex disease.

Meta-analysis identifies 13 new loci associated with waist-hip ratio and reveals sexual dimorphism in the genetic basis of fat distribution.

Waist-hip ratio (WHR) is a measure of body fat distribution and a predictor of metabolic consequences independent of overall adiposity. WHR is heritable, but few genetic variants influencing this trait have been identified. We conducted a meta-analysis of 32 genome-wide association studies for WHR adjusted for body mass index (comprising up to 77,167 participants), following up 16 loci in an additional 29 studies (comprising up to 113,636 subjects). We identified 13 new loci in or near RSPO3, VEGFA, TBX15-WARS2, NFE2L3, GRB14, DNM3-PIGC, ITPR2-SSPN, LY86, HOXC13, ADAMTS9, ZNRF3-KREMEN1, NISCH-STAB1 and CPEB4 (P = 1.9 × 10⁻⁹ to P = 1.8 × 10⁻⁴⁰) and the known signal at LYPLAL1. Seven of these loci exhibited marked sexual dimorphism, all with a stronger effect on WHR in women than men (P for sex difference = 1.9 × 10⁻³ to P = 1.2 × 10⁻&supl;³). These findings provide evidence for multiple loci that modulate body fat distribution independent of overall adiposity and reveal strong gene-by-sex interactions.

Genome-wide association and genetic functional studies identify autism susceptibility candidate 2 gene (AUTS2) in the regulation of alcohol consumption.

Alcohol consumption is a moderately heritable trait, but the genetic basis in humans is largely unknown, despite its clinical and societal importance. We report a genome-wide association study meta-analysis of ∼2.5 million directly genotyped or imputed SNPs with alcohol consumption (gram per day per kilogram body weight) among 12 population-based samples of European ancestry, comprising 26,316 individuals, with replication genotyping in an additional 21,185 individuals. SNP rs6943555 in autism susceptibility candidate 2 gene (AUTS2) was associated with alcohol consumption at genome-wide significance (P = 4 × 10(-8) to P = 4 × 10(-9)). We found a genotype-specific expression of AUTS2 in 96 human prefrontal cortex samples (P = 0.026) and significant (P < 0.017) differences in expression of AUTS2 in whole-brain extracts of mice selected for differences in voluntary alcohol consumption. Down-regulation of an AUTS2 homolog caused reduced alcohol sensitivity in Drosophila (P < 0.001). Our finding of a regulator of alcohol consumption adds knowledge to our understanding of genetic mechanisms influencing alcohol drinking behavior.

Genetic variants in novel pathways influence blood pressure and cardiovascular disease risk.

Blood pressure is a heritable trait influenced by several biological pathways and responsive to environmental stimuli. Over one billion people worldwide have hypertension (≥140 mm Hg systolic blood pressure or  ≥90 mm Hg diastolic blood pressure). Even small increments in blood pressure are associated with an increased risk of cardiovascular events. This genome-wide association study of systolic and diastolic blood pressure, which used a multi-stage design in 200,000 individuals of European descent, identified sixteen novel loci: six of these loci contain genes previously known or suspected to regulate blood pressure (GUCY1A3-GUCY1B3, NPR3-C5orf23, ADM, FURIN-FES, GOSR2, GNAS-EDN3); the other ten provide new clues to blood pressure physiology. A genetic risk score based on 29 genome-wide significant variants was associated with hypertension, left ventricular wall thickness, stroke and coronary artery disease, but not kidney disease or kidney function. We also observed associations with blood pressure in East Asian, South Asian and African ancestry individuals. Our findings provide new insights into the genetics and biology of blood pressure, and suggest potential novel therapeutic pathways for cardiovascular disease prevention.

Novel loci for adiponectin levels and their influence on type 2 diabetes and metabolic traits: a multi-ethnic meta-analysis of 45,891 individuals.

Circulating levels of adiponectin, a hormone produced predominantly by adipocytes, are highly heritable and are inversely associated with type 2 diabetes mellitus (T2D) and other metabolic traits. We conducted a meta-analysis of genome-wide association studies in 39,883 individuals of European ancestry to identify genes associated with metabolic disease. We identified 8 novel loci associated with adiponectin levels and confirmed 2 previously reported loci (P = 4.5×10(-8)-1.2×10(-43)). Using a novel method to combine data across ethnicities (N = 4,232 African Americans, N = 1,776 Asians, and N = 29,347 Europeans), we identified two additional novel loci. Expression analyses of 436 human adipocyte samples revealed that mRNA levels of 18 genes at candidate regions were associated with adiponectin concentrations after accounting for multiple testing (p<3×10(-4)). We next developed a multi-SNP genotypic risk score to test the association of adiponectin decreasing risk alleles on metabolic traits and diseases using consortia-level meta-analytic data. This risk score was associated with increased risk of T2D (p = 4.3×10(-3), n = 22,044), increased triglycerides (p = 2.6×10(-14), n = 93,440), increased waist-to-hip ratio (p = 1.8×10(-5), n = 77,167), increased glucose two hours post oral glucose tolerance testing (p = 4.4×10(-3), n = 15,234), increased fasting insulin (p = 0.015, n = 48,238), but with lower in HDL-cholesterol concentrations (p = 4.5×10(-13), n = 96,748) and decreased BMI (p = 1.4×10(-4), n = 121,335). These findings identify novel genetic determinants of adiponectin levels, which, taken together, influence risk of T2D and markers of insulin resistance.

A genome-wide association search for type 2 diabetes genes in African Americans.

African Americans are disproportionately affected by type 2 diabetes (T2DM) yet few studies have examined T2DM using genome-wide association approaches in this ethnicity. The aim of this study was to identify genes associated with T2DM in the African American population. We performed a Genome Wide Association Study (GWAS) using the Affymetrix 6.0 array in 965 African-American cases with T2DM and end-stage renal disease (T2DM-ESRD) and 1029 population-based controls. The most significant SNPs (n = 550 independent loci) were genotyped in a replication cohort and 122 SNPs (n = 98 independent loci) were further tested through genotyping three additional validation cohorts followed by meta-analysis in all five cohorts totaling 3,132 cases and 3,317 controls. Twelve SNPs had evidence of association in the GWAS (P<0.0071), were directionally consistent in the Replication cohort and were associated with T2DM in subjects without nephropathy (P<0.05). Meta-analysis in all cases and controls revealed a single SNP reaching genome-wide significance (P<2.5×10(-8)). SNP rs7560163 (P = 7.0×10(-9), OR (95% CI) = 0.75 (0.67-0.84)) is located intergenically between RND3 and RBM43. Four additional loci (rs7542900, rs4659485, rs2722769 and rs7107217) were associated with T2DM (P<0.05) and reached more nominal levels of significance (P<2.5×10(-5)) in the overall analysis and may represent novel loci that contribute to T2DM. We have identified novel T2DM-susceptibility variants in the African-American population. Notably, T2DM risk was associated with the major allele and implies an interesting genetic architecture in this population. These results suggest that multiple loci underlie T2DM susceptibility in the African-American population and that these loci are distinct from those identified in other ethnic populations.

Blood pressure loci identified with a gene-centric array.

Raised blood pressure (BP) is a major risk factor for cardiovascular disease. Previous studies have identified 47 distinct genetic variants robustly associated with BP, but collectively these explain only a few percent of the heritability for BP phenotypes. To find additional BP loci, we used a bespoke gene-centric array to genotype an independent discovery sample of 25,118 individuals that combined hypertensive case-control and general population samples. We followed up four SNPs associated with BP at our p < 8.56 × 10(-7) study-specific significance threshold and six suggestively associated SNPs in a further 59,349 individuals. We identified and replicated a SNP at LSP1/TNNT3, a SNP at MTHFR-NPPB independent (r(2) = 0.33) of previous reports, and replicated SNPs at AGT and ATP2B1 reported previously. An analysis of combined discovery and follow-up data identified SNPs significantly associated with BP at p < 8.56 × 10(-7) at four further loci (NPR3, HFE, NOS3, and SOX6). The high number of discoveries made with modest genotyping effort can be attributed to using a large-scale yet targeted genotyping array and to the development of a weighting scheme that maximized power when meta-analyzing results from samples ascertained with extreme phenotypes, in combination with results from nonascertained or population samples. Chromatin immunoprecipitation and transcript expression data highlight potential gene regulatory mechanisms at the MTHFR and NOS3 loci. These results provide candidates for further study to help dissect mechanisms affecting BP and highlight the utility of studying SNPs and samples that are independent of those studied previously even when the sample size is smaller than that in previous studies.

Discovery and refinement of loci associated with lipid levels.

Levels of low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides and total cholesterol are heritable, modifiable risk factors for coronary artery disease. To identify new loci and refine known loci influencing these lipids, we examined 188,577 individuals using genome-wide and custom genotyping arrays. We identify and annotate 157 loci associated with lipid levels at P < 5 × 10(-8), including 62 loci not previously associated with lipid levels in humans. Using dense genotyping in individuals of European, East Asian, South Asian and African ancestry, we narrow association signals in 12 loci. We find that loci associated with blood lipid levels are often associated with cardiovascular and metabolic traits, including coronary artery disease, type 2 diabetes, blood pressure, waist-hip ratio and body mass index. Our results demonstrate the value of using genetic data from individuals of diverse ancestry and provide insights into the biological mechanisms regulating blood lipids to guide future genetic, biological and therapeutic research.

New gene functions in megakaryopoiesis and platelet formation.

Platelets are the second most abundant cell type in blood and are essential for maintaining haemostasis. Their count and volume are tightly controlled within narrow physiological ranges, but there is only limited understanding of the molecular processes controlling both traits. Here we carried out a high-powered meta-analysis of genome-wide association studies (GWAS) in up to 66,867 individuals of European ancestry, followed by extensive biological and functional assessment. We identified 68 genomic loci reliably associated with platelet count and volume mapping to established and putative novel regulators of megakaryopoiesis and platelet formation. These genes show megakaryocyte-specific gene expression patterns and extensive network connectivity. Using gene silencing in Danio rerio and Drosophila melanogaster, we identified 11 of the genes as novel regulators of blood cell formation. Taken together, our findings advance understanding of novel gene functions controlling fate-determining events during megakaryopoiesis and platelet formation, providing a new example of successful translation of GWAS to function.

Common variants associated with plasma triglycerides and risk for coronary artery disease.

Triglycerides are transported in plasma by specific triglyceride-rich lipoproteins; in epidemiological studies, increased triglyceride levels correlate with higher risk for coronary artery disease (CAD). However, it is unclear whether this association reflects causal processes. We used 185 common variants recently mapped for plasma lipids (P < 5 × 10(-8) for each) to examine the role of triglycerides in risk for CAD. First, we highlight loci associated with both low-density lipoprotein cholesterol (LDL-C) and triglyceride levels, and we show that the direction and magnitude of the associations with both traits are factors in determining CAD risk. Second, we consider loci with only a strong association with triglycerides and show that these loci are also associated with CAD. Finally, in a model accounting for effects on LDL-C and/or high-density lipoprotein cholesterol (HDL-C) levels, the strength of a polymorphism's effect on triglyceride levels is correlated with the magnitude of its effect on CAD risk. These results suggest that triglyceride-rich lipoproteins causally influence risk for CAD.

Naamion takaa : näyttelijäntyön palapelini

The subject of this bachelor's thesis has its origins in the frequently asked question of an author; whether any of their own theatrical techniques correlate with those of any well-known theatrical personality. The aim of this bachelor's thesis, consequently, was to find out the true nature of the author as an actor and theatrical professional. The author started to put together a puzzle, from different fields of acting techniques, and came up with a solid picture. Different acting techniques and the basics of them were clarified and tested in practice. Techniques were reflected on, through great personalities in the field of theatre, via discussion. The selected personalities were Jouko Turkka, Antonin Artaud, Konstantin Stanislavski, Kaisa Korhonen and Bertolt Brecht. The toughness and physicality of acting played a strong role in this thesis. "Black pedagogy" was strongly criticized. The methods of Jouko Turkka were thoroughly investigated and, surprisingly to the author, turned out usable, although the author questioned the brutal ways that Turkka used them. The author carried out some of the physical exercises that corresponded to the different acting methods, and did "human tests" to understand the true meaning of the methods. Antonin Artaud abandoned all of his past life and sacrificed himself to art, which the author refuses to even consider it. The author found a soul-mate in Kaisa Korhonen and agreed with most of her methods. Many people who had worked with Kaisa Korhonen were interviewed to find out if she actually used the techniques. The author debated objectivity with Bertolt Brecht but, whereas Brecht saw it as the basis of everything, the author couldn't even consider it a usable technique. The author also had a discussion with Konstantin Stanislavski concerning the basics of acting, the study of singing, dancing and oral expression, and other beneficial skills. In the end, the author put herself on a plate, chopped herself into tiny pieces, started to study the pieces, and found out how she became as she is.

Esite rokotesuojauksesta ja terveysriskien ennaltaehkäisystä Aasiaan ja Etelä-Amerikkaan matkustaville virolaisille

Tämä opinnäytetyö liittyy Helsingin ammattikorkeakoulu Stadian ja Länsi-Tallinnan sairaalaan yhteiseen STaLT-projektiin. Opinnäytetyön tarkoituksena oli tuottaa yhteistyökumppanin toivomuksesta esite rokotesuojauksesta ja terveysriskien ennaltaehkäisystä Aasiaan ja Etelä- Amerikkaan matkustaville virolaisille matkailijoille. Ensimmäisenä tavoitteena oli kerätä tietoa matkailusta ja virolaisten matkailutottumuksista, terveysviestinnästä ja matkailijan terveysneuvonnasta. Erityisesti kartoitettiin tutkittua tietoa matkailijoiden terveysriskeistä ja näiden ennaltaehkäisevistä toimenpiteistä kuten rokotesuojauksesta. Toisena tavoitteena oli koota yhteenveto yleisimmistä matkailijaa kohtaavista tartuntataudeista sekä niiden ennaltaehkäisystä painottuen rokotesuojaukseen. Kolmantena tavoitteena oli suunnitella esite, josta hyödyn saavat etenkin Aasiaan ja Etelä-Amerikkaan matkustavat virolaiset sekä soveltuvin osin myös muut virolaiset matkailijat. Opinnäytetyö koostuu teoriaosuudesta sekä esitteestä. Teoriaosuus koostuu kirjallisuudesta ja tutkimuksista löytyvästä aineistosta, jonka perusteella työstimme matkailijoiden rokotesuojauksesta ja terveysriskien ennaltaehkäisystä esitteen, jota yhteistyökumppani voi käyttää oheismateriaalina toteuttaessaan terveysneuvontaa virolaisille matkailijoille. Kansainvälisen matkailun nopea kasvu nopeuttaa tartuntatautien leviämistä maailmassa. Laadukkaalla matkailijan terveysneuvonnalla pystytään ehkäisemään matkusteluun liittyviä terveysriskejä.

Seksuaalisuus ja ihmissuhteet

Toiminnallisen opinnäytetyömme tarkoituksena oli järjestää kehitysvammaisille nuorille Seksuaalisuus ja ihmissuhteet-kurssi yhteistyössä Helsingin Kehitysvammatuki 57 ry:n kanssa. Kurssi tarjosi osallistujille tukea ja ohjausta ihmissuhteisiin ja seksuaalisuuteen liittyvissä kysymyksissä, minkä tavoitteena oli lisätä nuorten sosiaalisia valmiuksia ja parantaa heidän kykyään tehdä itsenäisiä päätöksiä. Opinnäytetyömme lähtökohtana olivat ihmissuhteiden merkitys ja Bengt Nirjen normalisaatioperiaate, jonka mukaan kehitysvammaisten tulisi saada elää samoin arkielämän ehdoin kuin muidenkin. Seksuaalisuuden ilmaiseminen ja tyydyttävien ihmissuhteiden luominen on tärkeä osa jokaisen ihmisen elämää. Seksuaalisuus ja ihmissuhteet-kurssi edisti normalisaatioperiaatteen toteutumista normaalin elämänkaaren, itsemääräämisoikeuden ja seksuaalisuuden mallien alueilla. Seksuaalisuus ja ihmissuhteetkurssi toteutettiin syksyllä 2006 ja se sisälsi seitsemän kontaktikertaa. Kurssilla oli kahdeksan iältään 16-26-vuotiasta osallistujaa. Työskentelymenetelminä käytettiin luovia menetelmiä ja pienryhmätyöskentelyä. Lisäksi hyödynsimme selkokielistä materiaalia. Kurssipalaute kerättiin osallistujilta keskustelun ja lähipiiriltä palautelomakkeen avulla. Saadun palautteen perusteella kurssi onnistui tavoitteiden mukaisesti ja seksuaalisuuteen liittyvien kurssien järjestämistä pidettiin tärkeänä myös tulevaisuudessa. Seksuaalisuuteen ja ihmissuhteisiin liittyville kursseille on suuri tarve, johon vastaaminen vaatii ennakkoluulottomuutta sekä kurssitoiminnan järjestäjiltä että kehitysvammaisten lähipiireiltä.

Uhkaava ennenaikainen synnytys: hoitotyön prosessi Naistenklinikan osastolla 42 kätilöiden kokemana

Opinnäytetyön tarkoituksena oli selvittää ja kuvata miten hoitotyön prosessi etenee uhkaavassa ennenaikaisessa synnytyksessä Naistenklinikan osastolla 42 kätilöiden näkökulmasta. Lisäksi selvitettiin, minkälaisia haasteita uhkaavan ennenaikaisen synnytyksen hoitotyön prosessissa ilmenee. Opinnäytetyön tutkimusaineisto koottiin haastattelemalla viittä Naistenklinikan osastolla 42 työskentelevää kätilönä. Haastattelut toteutettiin teemahaastatteluina. Teemahaastatteluiden avulla pyrittiin saamaan vastauksia tutkimusongelmiin, jotka oli laadittu perehtymällä aihetta käsitteleviin tutkimuksiin ja kirjallisuuteen. Saatu tutkimusaineisto analysoitiin induktiivisen sisällönanalyysin avulla. Tulosten mukaan uhkaavan ennenaikaisen synnytyksen hoidossa kätilön rooli korostuu potilaan kokonaistilanteen kartoittamisessa sekä äidin fyysisen, psyykkisen ja sosiaalisen hyvinvoinnin tukemisessa. Osastolla toteutettavaa hoitoa ohjaavat lääkärin antamat yksilölliset hoito-ohjeet ja määräykset, mutta kätilöillä on myös oma hoitotyön näkökulma työskentelyssään äitien kanssa. Kätilötyön näkökulmasta uhkaavan ennenaikaisen synnytyksen hoito koostuu äidin ja sikiön fyysisen hyvinvoinnin sekä äidin psyykkisen ja sosiaalisen hyvinvoinnin tukemisesta. Fyysisen hyvinvoinnin tukemiseen kuuluvat lepohoidosta ja lääkehoidosta huolehtiminen, supistusten, vuotojen ja infektio-oireiden seuranta sekä sikiön voinnin tarkkailu. Psyykkisestä ja sosiaalisesta hyvinvoinnista huolehditaan pyrkimällä antamaan äideille riittävästi tukea, tietoa ja ohjausta sekä huomioimalla myös potilaan muun perheen jaksaminen. Uhkaavan ennenaikaisen synnytyksen hoito on moniammatillista yhteistyötä, jossa kätilö toimii hoitotiimin koordinaattorina. Haasteellisimmaksi asiaksi hoitotyössä haastatteluihin osallistuneet kätilöt nimesivät äitien psyykkisen hyvinvoinnin tukemisen. Tämän opinnäytetyön tutkimustuloksia voidaan hyödyntää uhkaavan ennenaikaisen synnytyksen hoitotyön prosessin arvioinnissa ja kehittämisessä Naistenklinikan osastolla 42. Lisäksi tätä opinnäytetyötä tullaan käyttämään lähteenä, kun osastolla 42 tehdään prosessikuvaus uhkaavan ennenaikaisen synnytyksen hoidosta ja hoitopolusta.