Interferència mediada per RNA, una nova revolució genètica

Els RNA (o ARN, àcids ribonucleics) són biomolècules lineals de cadena senzilla, com un fil, formades per la unió seqüencial d'altres molècules més senzilles, els nucleòtids. Abans de la descoberta del fenòmen de RNAi es creia que el RNA era només un intermediari silenciós de la maquinària genètica, que transportava cegament les instruccions dels gens, en descodificava el missatge i el convertia en proteïnes, procés que es coneix amb el nom de flux d'informació genètica (del gen, que emmagatzema la informació i és format per ADN, a les proteïnes, que fan la feina especificada pel gen) [...].

Clinical usefulness of the Screen for Cognitive Impairment in Psychiatry (SCIP-S) scale in patients with type I bipolar disorder

Background: The relevance of persistent cognitive deficits to the pathogenesis and prognosis of bipolar disorders (BD) is understudied, and its translation into clinical practice has been limited by the absence of brief methods assessing cognitive status in Psychiatry. This investigation assessed the psychometric properties of the Spanish version of the Screen for Cognitive Impairment in Psychiatry (SCIP-S) for the detection of cognitive impairment in BD. Methods: After short training, psychiatrists at 40 outpatient clinics administered the SCIP three times over two weeks to a total of 76 consecutive type I BD admissions. Experienced psychologists also administered a comprehensive battery of standard neuropsychological instruments to clinical sample and 45 healthy control subjects. Results: Feasibility was supported by a brief administration time (approximately 15 minutes) and minimal scoring errors. The reliability of the SCIP was confirmed by good equivalence of forms, acceptable stability (ICC range 0.59 to 0.87) and adequate internal consistency (Chronbach's alpha of 0.74). Construct validity was granted by extraction of a single factor (accounting 52% of the variance), acceptable correlations with conventional neuropsychological instruments, and a clear differentiation between bipolar I and normal samples. Efficiency was also provided by the adequate sensitivity and specificity. Limitations: The sample size is not very large. The SCIP and the neurocognitive battery do not cover all potentially relevant cognitive domains. Also, sensitivity to change remains unexplored. Conclusion: With minimal training, physicians obtained a reliable and valid estimate of cognitive impairment in approximately 15 minutes from an application of the SCIP to type I BD patients.

Clinical usefulness of the Screen for Cognitive Impairment in Psychiatry (SCIP-S) scale in patients with type I bipolar disorder

Background: The relevance of persistent cognitive deficits to the pathogenesis and prognosis of bipolar disorders (BD) is understudied, and its translation into clinical practice has been limited by the absence of brief methods assessing cognitive status in Psychiatry. This investigation assessed the psychometric properties of the Spanish version of the Screen for Cognitive Impairment in Psychiatry (SCIP-S) for the detection of cognitive impairment in BD. Methods: After short training, psychiatrists at 40 outpatient clinics administered the SCIP three times over two weeks to a total of 76 consecutive type I BD admissions. Experienced psychologists also administered a comprehensive battery of standard neuropsychological instruments to clinical sample and 45 healthy control subjects. Results: Feasibility was supported by a brief administration time (approximately 15 minutes) and minimal scoring errors. The reliability of the SCIP was confirmed by good equivalence of forms, acceptable stability (ICC range 0.59 to 0.87) and adequate internal consistency (Chronbach's alpha of 0.74). Construct validity was granted by extraction of a single factor (accounting 52% of the variance), acceptable correlations with conventional neuropsychological instruments, and a clear differentiation between bipolar I and normal samples. Efficiency was also provided by the adequate sensitivity and specificity. Limitations: The sample size is not very large. The SCIP and the neurocognitive battery do not cover all potentially relevant cognitive domains. Also, sensitivity to change remains unexplored. Conclusion: With minimal training, physicians obtained a reliable and valid estimate of cognitive impairment in approximately 15 minutes from an application of the SCIP to type I BD patients.

”Käsitteitä tarpeen mukaan". Kirjallisuustieteelliset käsitteet lukion kirjallisuudenopetuksessa

Tavoitteeni tässä tutkimuksessa oli selvittää, miten kirjallisuustieteellisiä proosa-anayysin käsitteitä opetetaan opetussuunnitelman mukaisissa lukion äidinkielen ja kirjallisuuden oppikirjoissa ja miten hyvin kokelaat hallitsivat kertoja-käsitteen tekstitaidon ylioppilaskokeessa keväällä 2007. Samalla pohdin, minkälainen kirjallisuustieteellinen käsitteistö palvelisi tekstianalyysin opetusta koulussa, koska Lukion opetussuunnitelman perusteet 2003 ja äidinkielen ylioppilaskoe edellyttävät äidinkielen ja kirjallisuuden opetukselta ja oppilailta käsitteiden käyttöä. Tutkimusaineistonani olivat kaikki kuusi käytössä olevaa lukion äidinkielen ja kirjallisuuden oppikirjaa ja 440 kpl kevään 2007 äidinkielen tekstitaidon ylioppilaskokeen vastaustekstiä. Oppikirjoja tarkastelin soveltamalla niiden arviointiin Lev S. Vygotskin ajatuksia arkikäsitteiden ja tieteellisten käsitteiden opettamisesta ja Hans Aeblin esittämiä teoreettisia malleja käsitteiden opettamisesta ja oppimisesta. Tutkimukseni osoittaa, että opetussuunnitelmassa mainittujen proosa-analyysin käsitteiden kertoja, näkökulma, motiivi, aihe ja teema opetus on epätäsmällistä. Oppikirjoissa ei ole otettu huomioon sitä, että käsitteenoppiminen on monivaiheinen prosessi. Myöskään problematiikkaa, joka aiheutuu kyseisten käsitteiden määrittelyn kirjavuudesta ja käytöstä sekä arkikielen käsitteinä että tieteellisinä käsitteinä, ei oppikirjoissa käsitellä. Sama näkyy ylioppilaskoeaineistossa: oppilaat eivät hallitse käsitettä kertoja tieteellisenä käsitteenä. Tietoisuus kirjallisuustieteellisten käsitteiden määrittelyn problematiikasta ja arkikäsitteiden ja tieteellisten käsitteiden ontologisista kategorioista on onnistuneen käsitteenoppimisen edellytys. Kirjallisuustieteelliset käsitteet ovat metakäsitteitä, jotka edellyttävät oppilaiden metakäsitteellisen tietoisuuden ja motivaation hyödyntämistä opetuksessa, jossa olisi sovellettava monipuolisesti eri oppimiskäsitysten parhaita puolia hyödyntäviä lähestymistapoja, erilaisia pedagogisia diskursseja. Koulujen kirjallisuudenopetusta suunniteltaessa ja kirjallisuustieteellisiä käsitteitä opetettaessa on otettava huomioon niin kirjallisuustieteen kuin kasvatustieteen näkökulma. Opetussuunnitelman ja ylioppilaskokeen asettama vaatimus käsitteiden käytöstä on kohtuuton, mikäli ei sovita, miten käsitteet määritellään ja mitä käsitteitä kokelaiden oletetaan ylioppilaskokeessa hallitsevan. Kirjallisuustieteellisten käsitteiden puutteellisen opetuksen oppikirjoissa ja niiden epämääräisen käytön ylioppilaskokeen tehtävänannoissa ja arvioinnissa voi kärjistyneimmillään nähdä oppilaan oikeusturvakysymyksenä

Hjärtats vanor, tankens välvilja och handens gärning : dygd som vårdetiskt grundbegrepp

The main aim of the study is to elucidate the meaning and dimensions of the concept of „virtue‟, and to find the place of virtue in a caritative caring ethics, i.e. a caring ethics based on human love and mercy. The intention is to create a theory model which utilizes the possibilities of virtue in developing the caritative caring ethics as a whole. The caritative caring ethics has a universal potential – it is primarily not a professional ethics, but it may form a frame of reference and basis for formulating ethical codes, and for ethical discussions within different caring contexts. The hermeneutic approach of the study is inspired by Gadamer‟s philosophical hermeneutics combined with the view of hermeneutics as a hypothetical-deductive process. The study is guided by Eriksson‟s model of definition of concepts. The concept of „virtue‟ is studied focusing on its ethical dimensions. These ethical dimensions of virtue are seen as anchored to an inner ethos, whereas ethos stands for the ontological goodness, a basic notion of the Good that permeates the entity of the human being, and forms the base of the culture where he lives and acts. The overarching research questions are: 1. What is virtue? 2. What is „virtue‟ as a basic concept in caring science? 3. What place does virtue have in caritative caring ethics? The answer of the first question is mainly searched for by an ontological determination comprising partly an etymologic and semantic analysis of „virtue‟, and partly a determination of the essence of virtue. The answer to the second and third questions are mainly searched for using a contextual determination, where the purposive context and pragmatic features of virtue are studied in relation to caring ethics. The ontological and contextual determinations are brought together through hermeneutical interpretation, forming a new whole, which constitutes the results of the study. The results of the study are depicted in a theory model, in which the movement of virtue from ethos to deed is moulded as caritative caring ethics. The material of the study consists of dictionaries, texts written by Aristotle and St. Thomas Aquinas, articles, dissertations, and books, as well as parts of a pilot survey answered by 33 nurses. The results of the study show that the essence of virtue is primarily functional, not ethical. The ethical emerges when virtue is contextualized in a human communion. Virtue makes something fulfil its function well; makes the human being good, and gives him morals and morality. The human being needs prudence, love, and humility to acquire and develop the moral virtues. Virtue is a power, related to a value, which considering a caritative caring ethics consists of the caritas motif. Human love is shown through deeds, making the human being do what he is expected to do. Virtue, as an active power of becoming, affirms and clarifies the human being‟s ability to develop in the direction of the Good. Virtue becomes essential and unifying when morality appears in the human mind as auctoritas, an inner, prompting power based on divinity or a transcendental ethos. Together ethos and virtue create opportunities for an inner ethics based on voluntariness and joy in being and doing the true, the good, and the beautiful.

Haljennut habitus : Bourdieun viimeisiksi jääneet omakohtaiset teokset englanninkielisinä käännöksinä

Kirjallisuusarvostelu

Identification and characterisation of a novel adhesin of Streptococcus suis and its use as a target of adhesion inhibition and bacterial detection

Streptococcus suis is an important pig pathogen but it is also zoonotic, i.e. capable of causing diseases in humans. Human S. suis infections are quite uncommon but potentially life-threatening and the pathogen is an emerging public health concern. This Gram-positive bacterium possesses a galabiose-specific (Galalpha1−4Gal) adhesion activity, which has been studied for over 20 years. P-fimbriated Escherichia coli−bacteria also possess a similar adhesin activity targeting the same disaccharide. The galabiose-specific adhesin of S. suis was identified by an affinity proteomics method. No function of the protein identified was formerly known and it was designated streptococcal adhesin P (SadP). The peptide sequence of SadP contains an LPXTG-motif and the protein was proven to be cell wall−anchored. SadP may be multimeric since in SDS-PAGE gel it formed a protein ladder starting from about 200 kDa. The identification was confirmed by producing knockout strains lacking functional adhesin, which had lost their ability to bind to galabiose. The adhesin gene was cloned in a bacterial expression host and properties of the recombinant adhesin were studied. The galabiose-binding properties of the recombinant protein were found to be consistent with previous results obtained studying whole bacterial cells. A live-bacteria application of surface plasmon resonance was set up, and various carbohydrate inhibitors of the galabiose-specific adhesins were studied with this assay. The potencies of the inhibitors were highly dependent on multivalency. Compared with P-fimbriated E. coli, lower concentrations of galabiose derivatives were needed to inhibit the adhesion of S. suis. Multivalent inhibitors of S. suis adhesion were found to be effective at low nanomolar concentrations. To specifically detect galabiose adhesin−expressing S. suis bacteria, a technique utilising magnetic glycoparticles and an ATP bioluminescence bacterial detection system was also developed. The identification and characterisation of the SadP adhesin give valuable information on the adhesion mechanisms of S. suis, and the results of this study may be helpful for the development of novel inhibitors and specific detection methods of this pathogen.

Receptors and endocytosis of coxsackievirus A9

Coxsackievirus A9 (CV-A9) belongs to human enteroviruses within family Picornaviridae, which are the main cause of aseptic meningitis. In addition, CV-A9 causes a wide range of other clinical manifestations of acute disease including respiratory infections, myocarditis, encephalitis and severe generalized infections in newborns. In this study, the functions of integrins αVβ6 and αVβ3 in the attachment and cellular entry of CV-A9 were analyzed. Further, virus and cell surface interactions and endocytosis of CV-A9 were studied in specific cell lines. Also, a method for production of GFP-expressing CV-A9 particles by long PCR-mediated mutagenesis and in vivo transcription was developed. The results indicated that RGD-motif (arginine-glycine-asparagine) that resides in the viral capsid is important for CV-A9 infection particularly in cell lines expressing integrin αVβ6 and that this integrin serves as a high affinity attachment receptor for the virus. CV-A9 is also capable of infecting certain cell lines independently of αV-integrins by binding to the cell surface HSPA5 protein. Regardless of the attachment stage, the internalization of the virus occurs via the same entry pathway and is dependent on β2M, dynamin, and Arf6 but independent of clathrin and caveolin-1. Furthermore, the virus internalization occurs within Arf6-containing vesicles suggesting that Arf6 is central mediator of CV-A9 endocytosis. While in this study the results of CV-A9 endocytosis were based on microscopical visualization within individual fixed cells, a rapid method for generation of a virus for real-time imaging was also described.

Aspectos clínicos, histopatológicos e moleculares da dermatosparaxia em ovinos White Dorper

Dermatosparaxia em animais é uma doença autossômica recessiva do tecido conjuntivo caracterizada por fragilidade e hiperextensibilidade cutânea. A doença em ovinos White Dorper é provocada pela mutação c.421G>T no gene ADAMmetalopeptidase com trombospondina tipo 1 motif, 2 (ADAMTS2). O objetivo deste estudo foi descrever os achados clínicos, moleculares e histopatológicos da dermatosparaxia em ovinos White Dorper de um rebanho localizado no Centro-Oeste Paulista. O rebanho era composto por nove animais, sendo um reprodutor, quatro matrizes e seus respectivos borregos. Dos nove animais examinados, dois apresentavam sinais clínicos compatíveis com dermatosparaxia. O exame histopatológico de amostras cutâneas das lesões destes dois animais revelou também achados compatíveis com dermatosparaxia, sendo caracterizados por epiderme e anexos cutâneos preservados e sem características atípicas; colágeno displásico arranjado em feixes pequenos, fragmentados e com focos de degeneração, anexos cutâneos proeminentes e na região da derme foco hemorrágico intenso associado a moderado infiltrado neutrofílico na derme profunda. Com o objetivo de realizar o diagnóstico molecular da enfermidade, uma PCR foi padronizada utilizando primers específicos desenhados para amplificar a região do gene ADAMTS2 que continha a mutação c.421G>T e o DNA obtido de amostras de sangue de todos os animais do rebanho. O sequenciamento direto dos produtos da PCR, comprovou que os dois animais clinicamente afetados possuíam a mutação responsável pela dermatosparaxia. A metodologia descrita neste estudo possibilitou o diagnóstico definitivo da doença. Segundo a literatura consultada, esta é a primeira vez que a dermatosparaxia é descrita em ovinos White Dorper no Brasil. A metodologia aqui descrita poderá ser empregada em estudos futuros que avaliem a prevalência desta mutação no Brasil, possibilitando a adoção de medidas que previnam a disseminação dessa mutação no rebanho brasileiro de ovinos White Dorper.

Prevalence study of SNP c.421G>T in the ADAMTS2 gene responsible for dermatosparaxis in White Dorper sheep in Brazil

Abstract: Dermatosparaxis is an autosomal recessive disorder of connective tissue; the disorder is clinically characterized by skin fragility and hyperextensibility. Dermatosparaxis in White Dorper sheep is caused by a single nucleotide polymorphism (SNP) (c.421G>T) in the ADAM metalloproteinase with thrombospondin type 1 motif, 2 (ADAMTS2) gene. The aim of this study was to investigate the prevalence of this SNP in a White Dorper herd in São Paulo state, Brazil. In this study, we collected blood DNA samples from 303 White Dorper sheep and performed polymerase chain reaction to amplify the SNP region. The samples were sequenced to determine the presence of the SNP in the ADAMTS2 gene. The SNP prevalence in the studied population was 15.5%; this finding indicates that more effective control measures should be used to prevent the inheritance of SNP c.421G>T in the ADAMTS2 gene in Brazilian White Dorper herds.

Palinologia de espécies da tribo Eupatorieae (Compositae Giseke) ocorrentes na Restinga de Carapebus, Carapebus, Rio de Janeiro

Foram estudados os grãos de pólen de sete táxons pertencentes à tribo Eupatorieae, ocorrentes na Restinga de Carapebus, Carapebus, Estado do Rio de Janeiro. São eles: Barrosoa atlantica King & Robins., B. betonicaeformis (DC.) King & Robins., Mikania belemii King & Robins., M. cordifolia Willd., M. glomerata Spreng., M. micrantha H.B.K., M. trinervis Hook & Arn. e Trichogoniopsis podocarpa (DC.) King & Robins. A tribo apresentou em comum, grãos de pólen pequenos a médios, oblato-esferoidais a prolato-esferoidais, tricolporados, sexina espinhosa e cavada. Os táxons puderam ser separados quando foram consideradas a forma polínica, as dimensões do espinho e a distância entre eles. Assim, foi possível formar dois conjuntos de espécies identificados pela forma polínica: o primeiro, com forma oblato-esferoidal, composto por Barrosoa atlantica, Mikania micrantha e M. trinervis e o segundo, com forma prolato-esferoidal, composto por Barrosoa betonicaeformis, Mikania belemii, M. cordifolia, M. glomerata e Trichogoniopsis podocarpa. Os resultados obtidos, em comparação à literatura corrente, permitem concluir que a tribo Eupatorieae é, palinologicamente, homogênea, porém algumas espécies podem ser separadas pelo grão de pólen, exceto M. glomerata de T. podocarpa e M. belemii de M. cordifolia.

Selaginellaceae Willk. no estado de São Paulo, Brasil

O presente trabalho trata do levantamento florístico da família Selaginellaceae Willk. no Estado de São Paulo. De acordo com os dados obtidos, foi possível o reconhecimento de 14 espécies nativas, distribuídas em três subgêneros: Heterostachys Baker, Stachygynandrum (P. Beauv.) Baker e Tetragonostachys Jermy. Os subgêneros Heterostachys e Tetragonostachys estão representados no Estado por uma única espécie cada, Selaginella muscosa Spring e Selaginella sellowii Hieron., respectivamente. O subgênero Stachygynandrum está representado por 12 espécies: Selaginella contigua Baker, S. convoluta (Arn.) Spring, S. decomposita Spring, S. flexuosa Spring, S. macrostachya (Spring) Spring, S. marginata (Humb. & Bonpl. ex Willd.) Spring, S. mendoncae Hieron., S. microphylla (Kunth) Spring, S. suavis (Spring) Spring, S. sulcata (Desv. ex Poir.) Spring, S. tenuissima Fée e S. valida Alston. Selaginella mendoncae e Selaginella sellowii estão sendo citadas pela primeira vez para o Estado de São Paulo. Além dessas 14 espécies nativas, também foram encontradas quatro espécies introduzidas - Selaginella kraussiana (Kunze) A. Braun, S. pallescens (C. Presl) Spring, S. plana (Desv. ex Poir.) Hieron. e S. vogelii Spring. São apresentadas chaves de identificação e descrições para os subgêneros e espécies, bem como ilustrações, distribuição geográfica e comentários das espécies estudadas.

Snake venom metalloproteinases and disintegrins: interactions with cells

Metalloproteinases and disintegrins are important components of most viperid and crotalid venoms. Large metalloproteinases referred to as MDC enzymes are composed of an N-terminal Metalloproteinase domain, a Disintegrin-like domain and a Cys-rich C-terminus. In contrast, disintegrins are small non-enzymatic RGD-containing cysteine-rich polypeptides. However, the disintegrin region of MDC enzymes bears a high degree of structural homology to that of the disintegrins, although it lacks the RGD motif. Despite these differences, both components share the property of being able to recognize integrin cell surface receptors and thereby to inhibit integrin-dependent cell reactions. Recently, several membrane-bound MDC enzymes, closely related to soluble venom MDC enzymes, have been described in mammalian cells. This group of membrane-anchored mammalian enzymes is also called the ADAM family of proteins due to the structure revealing A Disintegrin And Metalloproteinase domains. ADAMs are involved in the shedding of molecules from the cell surface, a property which is also shared by some venom MDC enzymes.

Purification and binding analysis of the nitrogen fixation regulatory NifA protein from Azospirillum brasilense

NifA protein activates transcription of nitrogen fixation operons by the alternative sigma54 holoenzyme form of RNA polymerase. This protein binds to a well-defined upstream activator sequence (UAS) located at the -200/-100 position of nif promoters with the consensus motif TGT-N10-ACA. NifA of Azospirillum brasilense was purified in the form of a glutathione-S-transferase (GST)-NifA fusion protein and proteolytic release of GST yielded inactive and partially soluble NifA. However, the purified NifA was able to induce the production of specific anti-A. brasilense NifA-antiserum that recognized NifA from A. brasilense but not from K. pneumoniae. Both GST-NifA and NifA expressed from the E. coli tac promoter are able to activate transcription from the nifHDK promoter but only in an A. brasilense background. In order to investigate the mechanism that regulates NifA binding capacity we have used E. coli total protein extracts expressing A. brasilense nifA in mobility shift assays. DNA fragments carrying the two overlapping, wild-type or mutated UAS motifs present in the nifH promoter region revealed a retarded band of related size. These data show that the binding activity present in the C-terminal domain of A. brasilense NifA protein is still functional even in the presence of oxygen.

Association of hepatic nuclear factor-4 in the apolipoprotein B promoter: a preliminary report

Previous studies have examined the arrangement of regulatory elements along the apolipoprotein B (apoB) promoter region (-3067 to +940) and a promoter fragment extending from nucleotides -150 to +124 has been demonstrated to be essential for transcriptional activation of the apoB gene in hepatic and intestinal cells. It has also been shown that transcriptional activation of apoB requires a synergistic interaction between hepatic nuclear factor-4 (HNF-4) and CCAAT/enhancer-binding protein a (C/EBPa) transcription factors. Here, we have examined the hypothesis that HNF-4 factor binding to DNA may induce a DNA helix bend, thus facilitating the communication with a C/EBPa factor located one helix turn from this HNF-4 factor in the apoB promoter. A gel electrophoretic mobility shift assay using wild type double-stranded oligonucleotides or modified wild type duplex oligonucleotides with 10 nucleotides inserted between HNF-4 and C/EBPa factor motifs showed similar retarded complexes, indicating that HNF-4 and C/EBPa factors interact independently of the distance between binding sites. However, when only one base, a thymidine, was inserted at the -71 position of the apoB promoter, the complex shift was completely abolished. In conclusion, these results regarding the study of the mechanisms involving the interaction between HNF-4 and C/EBPa factors in the apoB promoter suggest that the perfect 5'-CCCTTTGGA-3' motif is needed in order to facilitate the interaction between the two factors.