Reproducibility of Vertebral Fracture Assessment Readings From Dual-energy X-ray Absorptiometry in Both a Population-based and Clinical Cohort: Cohen's and Uniform Kappa.

Vertebral fracture assessments (VFAs) using dual-energy X-ray absorptiometry increase vertebral fracture detection in clinical practice and are highly reproducible. Measures of reproducibility are dependent on the frequency and distribution of the event. The aim of this study was to compare 2 reproducibility measures, reliability and agreement, in VFA readings in both a population-based and a clinical cohort. We measured agreement and reliability by uniform kappa and Cohen's kappa for vertebral reading and fracture identification: 360 VFAs from a population-based cohort and 85 from a clinical cohort. In the population-based cohort, 12% of vertebrae were unreadable. Vertebral fracture prevalence ranged from 3% to 4%. Inter-reader and intrareader reliability with Cohen's kappa was fair to good (0.35-0.71 and 0.36-0.74, respectively), with good inter-reader and intrareader agreement by uniform kappa (0.74-0.98 and 0.76-0.99, respectively). In the clinical cohort, 15% of vertebrae were unreadable, and vertebral fracture prevalence ranged from 7.6% to 8.1%. Inter-reader reliability was moderate to good (0.43-0.71), and the agreement was good (0.68-0.91). In clinical situations, the levels of reproducibility measured by the 2 kappa statistics are concordant, so that either could be used to measure agreement and reliability. However, if events are rare, as in a population-based cohort, we recommend evaluating reproducibility using the uniform kappa, as Cohen's kappa may be less accurate.

Mesoscale numerical analysis of the historical November 1982 heavy precipitation event over Andorra (Eastern Pyrenees)

From 6 to 8 November 1982 one of the most catastrophic flash-flood events was recorded in the Eastern Pyrenees affecting Andorra and also France and Spain with rainfall accumulations exceeding 400 mm in 24 h, 44 fatalities and widespread damage. This paper aims to exhaustively document this heavy precipitation event and examines mesoscale simulations performed by the French Meso-NH non-hydrostatic atmospheric model. Large-scale simulations show the slow-evolving synoptic environment favourable for the development of a deep Atlantic cyclone which induced a strong southerly flow over the Eastern Pyrenees. From the evolution of the synoptic pattern four distinct phases have been identified during the event. The mesoscale analysis presents the second and the third phase as the most intense in terms of rainfall accumulations and highlights the interaction of the moist and conditionally unstable flows with the mountains. The presence of a SW low level jet (30 m s-1) around 1500 m also had a crucial role on focusing the precipitation over the exposed south slopes of the Eastern Pyrenees. Backward trajectories based on Eulerian on-line passive tracers indicate that the orographic uplift was the main forcing mechanism which triggered and maintained the precipitating systems more than 30 h over the Pyrenees. The moisture of the feeding flow mainly came from the Atlantic Ocean (7-9 g kg-1) and the role of the Mediterranean as a local moisture source was very limited (2-3 g kg-1) due to the high initial water vapour content of the parcels and the rapid passage over the basin along the Spanish Mediterranean coast (less than 12 h).

The impact of adopting a CLIL approach on EFL learners reading skills in Catalonia school

En els últims anys, un gran nombre de docents i escoles han estat introduint programes AICLE a les escoles catalanes. A la majoria d’escoles, però, només s'ensenya anglès com a llengua estrangera, la qual cosa, limita les oportunitats d'estar en contacte amb la llengua estrangera en un context diferent. L’objectiu d'aquest estudi és analitzar si els infants milloren en les seves habilitats lectores i comprovar si existeix un avantatge significatiu respecte els alumnes que només estudien anglès com a llengua estrangera. L’estudi empíric que us presento a continuació, compara els resultats obtinguts en dues proves de comprensió lectora. A les proves hi van participar dos grups d’alumnes de cinquè de d’Educació Primària. Un dels grups seguia un programa AICLE (1 hora a la setmana i , a més aprenia anglès coma llengua estrangera); l’altre grup només estudiava anglès com a llengua estrangera. L'objectiu principal d'aquest projecte d'investigació és esbrinar si els programes AICLE tenen un impacte en les habilitats lectores en llengua anglesa dels alumnes. Així doncs, humilment intentaré contribuir a la literatura existent sobre els beneficis de l’AICLE.

Safety and immunogenicity of a chimpanzee adenovirus-vectored Ebola vaccine in healthy adults: a randomised, double-blind, placebo-controlled, dose-finding, phase 1/2a study.

BACKGROUND: The ongoing Ebola outbreak led to accelerated efforts to test vaccine candidates. On the basis of a request by WHO, we aimed to assess the safety and immunogenicity of the monovalent, recombinant, chimpanzee adenovirus type-3 vector-based Ebola Zaire vaccine (ChAd3-EBO-Z). METHODS: We did this randomised, double-blind, placebo-controlled, dose-finding, phase 1/2a trial at the Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland. Participants (aged 18-65 years) were randomly assigned (2:2:1), via two computer-generated randomisation lists for individuals potentially deployed in endemic areas and those not deployed, to receive a single intramuscular dose of high-dose vaccine (5 × 10(10) viral particles), low-dose vaccine (2·5 × 10(10) viral particles), or placebo. Deployed participants were allocated to only the vaccine groups. Group allocation was concealed from non-deployed participants, investigators, and outcome assessors. The safety evaluation was not masked for potentially deployed participants, who were therefore not included in the safety analysis for comparison between the vaccine doses and placebo, but were pooled with the non-deployed group to compare immunogenicity. The main objectives were safety and immunogenicity of ChAd3-EBO-Z. We did analysis by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT02289027. FINDINGS: Between Oct 24, 2014, and June 22, 2015, we randomly assigned 120 participants, of whom 18 (15%) were potentially deployed and 102 (85%) were non-deployed, to receive high-dose vaccine (n=49), low-dose vaccine (n=51), or placebo (n=20). Participants were followed up for 6 months. No vaccine-related serious adverse events were reported. We recorded local adverse events in 30 (75%) of 40 participants in the high-dose group, 33 (79%) of 42 participants in the low-dose group, and five (25%) of 20 participants in the placebo group. Fatigue or malaise was the most common systemic adverse event, reported in 25 (62%) participants in the high-dose group, 25 (60%) participants in the low-dose group, and five (25%) participants in the placebo group, followed by headache, reported in 23 (57%), 25 (60%), and three (15%) participants, respectively. Fever occurred 24 h after injection in 12 (30%) participants in the high-dose group and 11 (26%) participants in the low-dose group versus one (5%) participant in the placebo group. Geometric mean concentrations of IgG antibodies against Ebola glycoprotein peaked on day 28 at 51 μg/mL (95% CI 41·1-63·3) in the high-dose group, 44·9 μg/mL (25·8-56·3) in the low-dose group, and 5·2 μg/mL (3·5-7·6) in the placebo group, with respective response rates of 96% (95% CI 85·7-99·5), 96% (86·5-99·5), and 5% (0·1-24·9). Geometric mean concentrations decreased by day 180 to 25·5 μg/mL (95% CI 20·6-31·5) in the high-dose group, 22·1 μg/mL (19·3-28·6) in the low-dose group, and 3·2 μg/mL (2·4-4·9) in the placebo group. 28 (57%) participants given high-dose vaccine and 31 (61%) participants given low-dose vaccine developed glycoprotein-specific CD4 cell responses, and 33 (67%) and 35 (69%), respectively, developed CD8 responses. INTERPRETATION: ChAd3-EBO-Z was safe and well tolerated, although mild to moderate systemic adverse events were common. A single dose was immunogenic in almost all vaccine recipients. Antibody responses were still significantly present at 6 months. There was no significant difference between doses for safety and immunogenicity outcomes. This acceptable safety profile provides a reliable basis to proceed with phase 2 and phase 3 efficacy trials in Africa. FUNDING: Swiss State Secretariat for Education, Research and Innovation (SERI), through the EU Horizon 2020 Research and Innovation Programme.

Perspectives interculturelles et interlinguistiques sur le discours académique – Crosscultural and Crosslinguistic Perspectives on Academic Discourse. Volume 1

The Department of French Studies of the University of Turku (Finland) organized an International Bilingual Conference on Crosscultural and Crosslinguistic Perspectives on Academic Discourse from 2022 May 2005. The event hosted specialists on Academic Discourse from Belgium, Finland, France, Germany, Italy, Norway, Spain, and the USA. This book is the first volume in our series of publications on Academic Discourse (AD hereafter). The following pages are composed of selected papers from the conference and focus on different aspects and analytical frameworks of Academic Discourse. One of the motivations behind organizing the conference was to examine and expand research on AD in different languages. Another one was to question to what extent academic genres are culturebound and language specific or primarily field or domain specific. The research carried out on AD has been mainly concerned with the use of English in different academic settings for a long time now – mainly written contexts – and at the expense of other languages. Alternatively the academic genre conventions of English and English speaking world have served as a basis for comparison with other languages and cultures. We consider this first volume to be a strong contribution to the spreading out of researches based on other languages than English in AD, namely Finnish, French, Italian, Norwegian and Romanian in this book. All the following articles have a strong link with the French language: either French is constitutive of the AD corpora under examination or the article was written in French. The structure of the book suggests and provides evidence that the concept of AD is understood and tackled to varying degrees by different scholars. Our first volume opens up the discussion on what AD is and backs dissemination, overlapping and expansion of current research questions and methodologies. The book is divided into three parts and contains four articles in English and six articles in French. The papers in part one and part two cover what we call the prototypical genre of written AD, i.e. the research article. Part one follows up on issues linked to the 13 Research Article (RA hereafter). Kjersti Fløttum asks wether a typical RA exists and concentrates on authors’ voices in RA (self and other dimensions), whereas Didriksen and Gjesdal’s article focuses on individual variation of the author’s voice in RA. The last article in this section is by Nadine Rentel and deals with evaluation in the writing of RA. Part two concentrates on the teaching and learning of AD within foreign language learning, another more or less canonical genre of AD. Two aspects of writing are covered in the first two articles: foreign students’ representations on rhetorical traditions (Hidden) and a contrastive assessment of written exercices in French and Finnish in Higher Education (Suzanne). The last contribution in this section on AD moves away from traditional written forms and looks at how argumentation is constructed in students’ oral presentations (Dervin and Fauveau). The last part of the book continues the extension by featuring four articles written in French exploring institutional and scientific discourses. Institutional discourses under scrutiny include the European Bologna Process (Galatanu) and Romanian reform texts (Moilanen). As for scientific discourses, the next paper in this section deconstructs an ideological discourse on the didactics of French as a foreign language (Pescheux). Finally, the last paper in part three reflects on varied forms of AD at university (Defays). We hope that this book will add some fuel to continue discussing diverse forms of and approches to AD – in different languages and voices! No need to say that with the current upsurge in academic mobility, reflecting on crosscultural and crosslinguistic AD has just but started.

Witness is presence : reading Stanley Hauerwas in a Nordic setting

I en postsekulär situation har förutsättningarna för att förstå och kommunicera en kristen tradition förändrats. De etablerade religionerna kan inte längre anta att de har monopol på den ”religiösa marknaden”. Ett centralt påstående i denna studie är att en postsekulär situation, som kännetecknas av minskande medlemskap i etablerade kyrkor och en ny synlighet av alternativa religiösa uttryck, medför ett behov av att reflektera över alternativa förståelser av kristenheten i sin kontext. Denna studie frågar hur man kan meningsfullt kommunicera en kristen tradition i en postsekulär situation. Traditionellt uttryckt: hur kan man förstå kristet vittnesbörd i vår situation? Det är till denna problematik som denna studie hävdar att Stanley Hauerwas ecklesiologi tillför ett viktigt perspektiv. Hauerwas ses som ett kritiskt korrektiv till vissa synsätt som ofta möter hos Nordiska, i denna studie främst finska och svenska, lutherska teologer. Det perspektiv som utvecklas i denna studie anses vara relevant eftersom kristna kyrkor i en postsekulär situation inte längre kan anta en makt- eller majoritetsposition. Därmed uppstår ett behov av att fråga sig hur man kan förstå kristenhet som en gemenskap i en minoritetsposition som bär vittnesbörd utan samhällelig makt. Denna studie hävdar att förkroppsligande av kristna övertygelser intar en central roll i en meningsfull postsekulär förståelse av vittnesbörd.

Building a creative city brand through an international mega-event : Case: Helsinki World Design Capital 2012

In order to grow, cities are increasingly competing for attention, jobs, investments, visitors, residents and significant events. Cities need to come up with creative solutions to keep up with the competition; they ought to become creative cities. Attracting talented and diverse inhabitants is a key factor in developing a creative city, which on is characterized by openness, tolerance, vibrancy and diversity. Along the need for renewed city images city brand building has become popular. Helsinki is the World Design Capital 2012 (WDC 2012) and this mega-event presents a meaningful opportunity for the city to broadcast itself globally. The purpose of this study is to evaluate how Helsinki brands itself as a creative city through an international mega-event. The sub-aims are to: 1) Map the factors behind the creative city and their relation to the city of Helsinki, 2) Describe the city branding process, 3) Evaluate the role of the Helsinki World Design Capital 2012 mega-event in Helsinki’s creative city brand building. First, the theory discusses the concept of the creative city that has gained growing attention during the past decade. Then, the city branding process is described and the benefits of hosting a mega-event are presented. Finally, co-branding a city and a mega-event in order to generate maximum benefit from the mega-event, is reviewed. This is a qualitative research for which data was collected through three face-to-face interviews, the World Design Capital 2012 bid, Helsinki’s economic development strategy, a consulting firm’s research report on the case city and web-pages. The research reveals that Helsinki has shown interest in the creative city discussion. The terminology around the concept is however approached carefully. Helsinki fits many of the creative city characteristics and recognizes its flaws for which improvement strategies have been planned. Bottlenecks keeping the city from promoting a more open mind were mainly revealed in its organizational structures. Helsinki has no official brand strategy; nonetheless pressure to develop one is present. The World Design Capital 2012 mega-event is seen as a meaningful stepping board to strengthen Helsinki’s identity and image, and start thinking about a city brand. The brand strategies of the mega-event support the values and virtues of the city itself, which enables benefits of co-branding introduces in the theory part. Helsinki has no official brand and doesn’t call itself a creative city, however this study shows signs of the city taking steps towards building a creative city brand with the help of the Helsinki World Design Capital 2012 mega-event.

Skill Development in Music Reading. The Eye-Movement Approach

This dissertation examined skill development in music reading by focusing on the visual processing of music notation in different music-reading tasks. Each of the three experiments of this dissertation addressed one of the three types of music reading: (i) sight-reading, i.e. reading and performing completely unknown music, (ii) rehearsed reading, during which the performer is already familiar with the music being played, and (iii) silent reading with no performance requirements. The use of the eye-tracking methodology allowed the recording of the readers’ eye movements from the time of music reading with extreme precision. Due to the lack of coherence in the smallish amount of prior studies on eye movements in music reading, the dissertation also had a heavy methodological emphasis. The present dissertation thus aimed to promote two major issues: (1) it investigated the eye-movement indicators of skill and skill development in sight-reading, rehearsed reading and silent reading, and (2) developed and tested suitable methods that can be used by future studies on the topic. Experiment I focused on the eye-movement behaviour of adults during their first steps of learning to read music notation. The longitudinal experiment spanned a nine-month long music-training period, during which 49 participants (university students taking part in a compulsory music course) sight-read and performed a series of simple melodies in three measurement sessions. Participants with no musical background were entitled as “novices”, whereas “amateurs” had had musical training prior to the experiment. The main issue of interest was the changes in the novices’ eye movements and performances across the measurements while the amateurs offered a point of reference for the assessment of the novices’ development. The experiment showed that the novices tended to sight-read in a more stepwise fashion than the amateurs, the latter group manifesting more back-and-forth eye movements. The novices’ skill development was reflected by the faster identification of note symbols involved in larger melodic intervals. Across the measurements, the novices also began to show sensitivity to the melodies’ metrical structure, which the amateurs demonstrated from the very beginning. The stimulus melodies consisted of quarter notes, making the effects of meter and larger melodic intervals distinguishable from effects caused by, say, different rhythmic patterns. Experiment II explored the eye movements of 40 experienced musicians (music education students and music performance students) during temporally controlled rehearsed reading. This cross-sectional experiment focused on the eye-movement effects of one-bar-long melodic alterations placed within a familiar melody. The synchronizing of the performance and eye-movement recordings enabled the investigation of the eye-hand span, i.e., the temporal gap between a performed note and the point of gaze. The eye-hand span was typically found to remain around one second. Music performance students demonstrated increased professing efficiency by their shorter average fixation durations as well as in the two examined eye-hand span measures: these participants used larger eye-hand spans more frequently and inspected more of the musical score during the performance of one metrical beat than students of music education. Although all participants produced performances almost indistinguishable in terms of their auditory characteristics, the altered bars indeed affected the reading of the score: the general effects of expertise in terms of the two eye- hand span measures, demonstrated by the music performance students, disappeared in the face of the melodic alterations. Experiment III was a longitudinal experiment designed to examine the differences between adult novice and amateur musicians’ silent reading of music notation, as well as the changes the 49 participants manifested during a nine-month long music course. From a methodological perspective, an opening to research on eye movements in music reading was the inclusion of a verbal protocol in the research design: after viewing the musical image, the readers were asked to describe what they had seen. A two-way categorization for verbal descriptions was developed in order to assess the quality of extracted musical information. More extensive musical background was related to shorter average fixation duration, more linear scanning of the musical image, and more sophisticated verbal descriptions of the music in question. No apparent effects of skill development were observed for the novice music readers alone, but all participants improved their verbal descriptions towards the last measurement. Apart from the background-related differences between groups of participants, combining verbal and eye-movement data in a cluster analysis identified three styles of silent reading. The finding demonstrated individual differences in how the freely defined silent-reading task was approached. This dissertation is among the first presentations of a series of experiments systematically addressing the visual processing of music notation in various types of music-reading tasks and focusing especially on the eye-movement indicators of developing music-reading skill. Overall, the experiments demonstrate that the music-reading processes are affected not only by “top-down” factors, such as musical background, but also by the “bottom-up” effects of specific features of music notation, such as pitch heights, metrical division, rhythmic patterns and unexpected melodic events. From a methodological perspective, the experiments emphasize the importance of systematic stimulus design, temporal control during performance tasks, and the development of complementary methods, for easing the interpretation of the eye-movement data. To conclude, this dissertation suggests that advances in comprehending the cognitive aspects of music reading, the nature of expertise in this musical task, and the development of educational tools can be attained through the systematic application of the eye-tracking methodology also in this specific domain.

Financial Slack and Bidding Firms’ Takeover Announcement Stock Returns – an Event Study

Despite the fact that the literature on mergers and acquisitions is extensive, relatively little effort has been made to examine the relationship between the acquiring firms’ financial slack and short-term post-takeover announcement abnormal stock returns. In this study, the case is made that the financial slack of a firm is not only an outcome of past business and financing activities but it also may affect the quality of acquisition decisions. We will hypothesize that the level of financial slack in a firm is negatively associated with the abnormal returns following acquisition announcements because slack reduces managerial discipline over the use of corporate funds and also because it may give rise to managerial self-serving behavior. In this study, financial slack is measured in terms of three financial statements ratios: leverage ratio, cash and equivalents to total assets ratio and free cash flow to total assets ratio. The data used in this paper is collected from two main sources. A list comprising 90 European acquisition announcements is retrieved from Thomson One Banker database. The stock price data and financial statements information for the respective firms is collected using Datastream. Our empirical analysis is two-fold. First, we conduct a two-sample t-test where we find that the most slack-rich firms experience lower abnormal returns than the most slack-poor firms in the event window [-1, +1], significant at 5% risk level. Second, we perform a cross sectional regression for sample firms using three financial statements ratios to explain cumulative abnormal returns (CAR). We find that leverage shows a statistically significant positive relationship with cumulative abnormal returns in event window [-1; +1] (significance 5%). Moreover, cash to total assets ratio showed a weak negative relationship with CAR (significant at 10%) in event window [-1; +1]. We conclude that our hypothesis for the inverse relationship between slack and abnormal returns receives empirical support. Based on the results of the event study we get empirical support for the hypothesis that the capital markets expect the acquisitions undertaken by slack-rich firms to more likely be driven by managerial self-serving behavior and hubris than do those undertaken by slackpoor firms, signaling possible agency problems and behavioral biases.

Elevers läs- och skrivförmåga över tid : en longitudinell kohortstudie av läsförståelse och stavning i årskurs 1-6 i en finlandssvensk skola

Reports have shown that a growing number of students are leaving school without adequate reading and writing skills. At the same time, finnish children have improved in reading lit-eracy over the past decade. The school´s primary mission is to support student´s reading and writing development and follow it over time. Not only individual students´development has to be followd over time, as seen in this study, to be able to plan interventions. The purpose of this licentiate thesis is to study how the results in grades 1–6 exhibit in reading compre-hension and spelling over time in a school for the Swedish-speaking minority in Finland. Even gender differences in reading comprehension and spelling are being examined, as well as students with low performance. The data collection has been done during a period of twelwe years, 1997–2009, where data from practically every two years has been used for this study. Results are reported for the years 1997, 2000, 2001, 2003, 2007 and 2009. The measuring instruments are standardized and normative tests in reading comprehension and spelling. The same instruments were used all of the years. The number of students was 1 037 and when possible, students partici-pated more than once. There was a minimal loss of data. The groups of students mostly came from urban districts, and a smaller number came from rural districts. Students with special needs were included in the group, but none of the students in special units participat-ed. The results show that the skills in reading comprehension and spelling haven´t changed appreciably over time. The trend shows slightly positive or unchanged direction except in spelling in grade 1 and reading comprehension in grade 5 where the trend is slightly nega-tive. The most obvious change can be noticed over time in reading comprehension in grade 2, where performance has improved significantly. Significant differences between boys´and girls´performances occur some years and in favor of the girls, but more often in spelling than in reading comprehension. In grade 4, there are no differences in results between boys and girls in any year, neither in reading comprehension nor in spelling, nor in grade 2 in reading comprehension. When the results shows gender differences, there are always differ-ences in both reading comprehension and spelling. Poor performances are also analyzed over time. Isolated reading comprehension difficulties were common for those who was performing poorly. In grade 2, as the only grade, most common was a double deficit in both reading comprehension and spelling, and in grade 4, most students who performed poorly had isolated difficulties in spelling. The amount of poor results varies over time, and no particular trend can be discerned. In grade 2, however, the poor results decreased over time, especially for the boys. Even in reading comprehension for grade 6 the amount of poor results, especially for the boys (10th percentile), has decreased over time, at least in the later years, while the poor results in-creased in spelling generally. In grade 4 the amount of poor results in spelling (10th perc.) seems to be reduced for the girls.

A comparative analysis of preprocessing techniques of cardiac event series for the study of heart rhythm variability using simulated signals

In the present study, using noise-free simulated signals, we performed a comparative examination of several preprocessing techniques that are used to transform the cardiac event series in a regularly sampled time series, appropriate for spectral analysis of heart rhythm variability (HRV). First, a group of noise-free simulated point event series, which represents a time series of heartbeats, was generated by an integral pulse frequency modulation model. In order to evaluate the performance of the preprocessing methods, the differences between the spectra of the preprocessed simulated signals and the true spectrum (spectrum of the model input modulating signals) were surveyed by visual analysis and by contrasting merit indices. It is desired that estimated spectra match the true spectrum as close as possible, showing a minimum of harmonic components and other artifacts. The merit indices proposed to quantify these mismatches were the leakage rate, defined as a measure of leakage components (located outside some narrow windows centered at frequencies of model input modulating signals) with respect to the whole spectral components, and the numbers of leakage components with amplitudes greater than 1%, 5% and 10% of the total spectral components. Our data, obtained from a noise-free simulation, indicate that the utilization of heart rate values instead of heart period values in the derivation of signals representative of heart rhythm results in more accurate spectra. Furthermore, our data support the efficiency of the widely used preprocessing technique based on the convolution of inverse interval function values with a rectangular window, and suggest the preprocessing technique based on a cubic polynomial interpolation of inverse interval function values and succeeding spectral analysis as another efficient and fast method for the analysis of HRV signals

Biphasic modulation of insulin receptor substrate-1 during goitrogenesis

Insulin receptor substrate-1 (IRS-1) is the main intracellular substrate for both insulin and insulin-like growth factor I (IGF-I) receptors and is critical for cell mitogenesis. Thyrotropin is able to induce thyroid cell proliferation through the cyclic AMP intracellular cascade; however, the presence of either insulin or IGF-I is required for the mitogenic effect of thyroid-stimulating hormone (TSH) to occur. The aim of the present study was to determine whether thyroid IRS-1 content is modulated by TSH in vivo. Strikingly, hypothyroid goitrous rats, which have chronically high serum TSH levels (control, C = 2.31 ± 0.28; methimazole (MMI) 21d = 51.02 ± 6.02 ng/mL, N = 12 rats), when treated with 0.03% MMI in drinking water for 21 days, showed significantly reduced thyroid IRS-1 mRNA content. Since goiter was already established in these animals by MMI for 21 days, we also evaluated IRS-1 expression during goitrogenesis. Animals treated with MMI for different periods of time showed a progressive increase in thyroid weight (C = 22.18 ± 1.21; MMI 5d = 32.83 ± 1.48; MMI 7d = 31.1 ± 3.25; MMI 10d = 33.8 ± 1.25; MMI 14d = 45.5 ± 2.56; MMI 18d = 53.0 ± 3.01; MMI 21d = 61.9 ± 3.92 mg, N = 9-15 animals per group) and serum TSH levels (C = 1.57 ± 0.2; MMI 5d = 9.95 ± 0.74; MMI 7d = 10.38 ± 0.84; MMI 10d = 17.72 ± 1.47; MMI 14d = 25.65 ± 1.23; MMI 18d = 35.38 ± 3.69; MMI 21d = 31.3 ± 2.7 ng/mL, N = 9-15 animals per group). Thyroid IRS-1 mRNA expression increased progressively during goitrogenesis, being significantly higher by the 14th day of MMI treatment, and then started to decline, reaching the lowest values by the 21st day, when a significant reduction was detected. In the liver of these animals, however, a significant decrease of IRS-1 mRNA was detected after 14 days of MMI treatment, a mechanism probably involved in the insulin resistance that occurs in hypothyroidism. The increase in IRS-1 expression during goitrogenesis may represent an important event associated with the increased rate of cell mitosis promoted by TSH and indicates that insulin and IGF-I are important co-mitogenic factors in vivo, possibly acting through the activation of IRS-1.

Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87% identity in amino acid sequence with Eur m 1 but only 80% with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96%), an extended strand (17.13%), a ß turn (5.61%), and a random coil (43.30%). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.