Polimorfismos de nucleotídeo simples (SNPs) em genes codificadores de citocinas e suas correlações com parâmetros clínicos e laboratoriais de pacientes portadores do vírus da imunodeficiência humana

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Astenia dérmica regional hereditária equina: diagnóstico, ocorrência no Brasil e caracterização clinica

Pós-graduação em Medicina Veterinária - FMVZ

Challenges for Brazilian MNCs' international human resources management

Multinationals from emerging countries such as Brazil now take the path of internationalisation where early movers have already been. However, these companies have to develop new tools to deal with their own challenges, since they come from countries with different historical backgrounds and specificities. This paper explores challenges for Brazilian MNCs in terms of HRM when operating abroad. It presents the results of six cases of Brazilian MNCs through a grounded theory study. Results show these companies had to deal with their former economic turbulence, shortage of qualified workforce to work internationally and the need to develop HRM competencies to operate globally.

Different approaches to identify markers for meat quality and production traits in pigs: analysis of the transcriptome in post mortem skeletal muscle and investigation of candidate genes

Due to the growing attention of consumers towards their food, improvement of quality of animal products has become one of the main focus of research. To this aim, the application of modern molecular genetics approaches has been proved extremely useful and effective. This innovative drive includes all livestock species productions, including pork. The Italian pig breeding industry is unique because needs heavy pigs slaughtered at about 160 kg for the production of high quality processed products. For this reason, it requires precise meat quality and carcass characteristics. Two aspects have been considered in this thesis: the application of the transcriptome analysis in post mortem pig muscles as a possible method to evaluate meat quality parameters related to the pre mortem status of the animals, including health, nutrition, welfare, and with potential applications for product traceability (chapters 3 and 4); the study of candidate genes for obesity related traits in order to identify markers associated with fatness in pigs that could be applied to improve carcass quality (chapters 5, 6, and 7). Chapter three addresses the first issue from a methodological point of view. When we considered this issue, it was not obvious that post mortem skeletal muscle could be useful for transcriptomic analysis. Therefore we demonstrated that the quality of RNA extracted from skeletal muscle of pigs sampled at different post mortem intervals (20 minutes, 2 hours, 6 hours, and 24 hours) is good for downstream applications. Degradation occurred starting from 48 h post mortem even if at this time it is still possible to use some RNA products. In the fourth chapter, in order to demonstrate the potential use of RNA obtained up to 24 hours post mortem, we present the results of RNA analysis with the Affymetrix microarray platform that made it possible to assess the level of expression of more of 24000 mRNAs. We did not identify any significant differences between the different post mortem times suggesting that this technique could be applied to retrieve information coming from the transcriptome of skeletal muscle samples not collected just after slaughtering. This study represents the first contribution of this kind applied to pork. In the fifth chapter, we investigated as candidate for fat deposition the TBC1D1 [TBC1 (tre-2/USP6, BUB2, cdc16) gene. This gene is involved in mechanisms regulating energy homeostasis in skeletal muscle and is associated with predisposition to obesity in humans. By resequencing a fragment of the TBC1D1 gene we identified three synonymous mutations localized in exon 2 (g.40A>G, g.151C>T, and g.172T>C) and 2 polymorphisms localized in intron 2 (g.219G>A and g.252G>A). One of these polymorphisms (g.219G>A) was genotyped by high resolution melting (HRM) analysis and PCR-RFLP. Moreover, this gene sequence was mapped by radiation hybrid analysis on porcine chromosome 8. The association study was conducted in 756 performance tested pigs of Italian Large White and Italian Duroc breeds. Significant results were obtained for lean meat content, back fat thickness, visible intermuscular fat and ham weight. In chapter six, a second candidate gene (tribbles homolog 3, TRIB3) is analyzed in a study of association with carcass and meat quality traits. The TRIB3 gene is involved in energy metabolism of skeletal muscle and plays a role as suppressor of adipocyte differentiation. We identified two polymorphisms in the first coding exon of the porcine TRIB3 gene, one is a synonymous SNP (c.132T> C), a second is a missense mutation (c.146C> T, p.P49L). The two polymorphisms appear to be in complete linkage disequilibrium between and within breeds. The in silico analysis of the p.P49L substitution suggests that it might have a functional effect. The association study in about 650 pigs indicates that this marker is associated with back fat thickness in Italian Large White and Italian Duroc breeds in two different experimental designs. This polymorphisms is also associated with lactate content of muscle semimembranosus in Italian Large White pigs. Expression analysis indicated that this gene is transcribed in skeletal muscle and adipose tissue as well as in other tissues. In the seventh chapter, we reported the genotyping results for of 677 SNPs in extreme divergent groups of pigs chosen according to the extreme estimated breeding values for back fat thickness. SNPs were identified by resequencing, literature mining and in silico database mining. analysis, data reported in the literature of 60 candidates genes for obesity. Genotyping was carried out using the GoldenGate (Illumina) platform. Of the analyzed SNPs more that 300 were polymorphic in the genotyped population and had minor allele frequency (MAF) >0.05. Of these SNPs, 65 were associated (P<0.10) with back fat thickness. One of the most significant gene marker was the same TBC1D1 SNPs reported in chapter 5, confirming the role of this gene in fat deposition in pig. These results could be important to better define the pig as a model for human obesity other than for marker assisted selection to improve carcass characteristics.

PTCH promoter methylation at low level in sporadic basal cell carcinoma analysed by three different approaches

Please cite this paper as: PTCH promoter methylation at low level in sporadic basal cell carcinoma analysed by three different approaches. Experimental Dermatology 2010. Abstract: Basal cell carcinoma (BCC) is the most common form of skin cancer. Mutations of the PTCH hallmark gene are detected in about 50-60% of BCCs, which raises the question whether other mechanisms such as promoter methylation can inactivate PTCH. Therefore, we performed methylation analysis of the PTCH promoter in a total of 56 BCCs. The sensitivity of three different methods, including direct bisulphite sequencing PCR, MethyLight and high-resolution melting (HRM), was applied and compared. We found that HRM analysis of DNA from fresh tissue [rather than formalin-fixed and paraffin-embedded tissue (FFPE)] was the most sensitive method to detect methylation. Low-level methylation of the PTCH promoter was detected in five out of 16 analysed BCCs (31%) on DNA from fresh tissue but only in two (13%) samples on short-time stored FFPE DNA from the very same tumors. In contrast, we were unable to detect methylation by HRM on long-time stored DNA in any of the remaining 40 BCC samples. Our data suggest that (i) HRM on DNA extracted from fresh tissue is the most sensitive method to detect methylation and (ii) methylation of the PTCH promoter may only play a minor role in BCC carcinogenesis.

Chicago classification criteria of esophageal motility disorders defined in high resolution esophageal pressure topography

The Chicago Classification of esophageal motility was developed to facilitate the interpretation of clinical high resolution esophageal pressure topography (EPT) studies, concurrent with the widespread adoption of this technology into clinical practice. The Chicago Classification has been an evolutionary process, molded first by published evidence pertinent to the clinical interpretation of high resolution manometry (HRM) studies and secondarily by group experience when suitable evidence is lacking.

Public service motivation and performance: a critical perspective

Purpose – A growing body of literature points to the importance of public service motivation (PSM) for the performance of public organizations. The purpose of this paper is to assess the method predominantly used for studying this linkage by comparing the findings it yields without and with a correction suggested by Brewer (2006), which removes the common-method bias arising from employee-specific response tendencies. Design/methodology/approach – First, the authors conduct a systematic review of published empirical research on the effects of PSM on performance and show that all studies found have been conducted at the individual level. Performance indicators in all but three studies were obtained by surveying the same employees who were also asked about their PSM. Second, the authors conduct an empirical analysis. Using survey data from 240 organizational units within the Swiss federal government, the paper compares results from an individual-level analysis (comparable to existing research) to two analyses where the data are aggregated to the organizational level, one without and one with the correction for common-method bias suggested by Brewer (2006). Findings – Looking at the Attraction to Policy-Making dimension of PSM, there is an interesting contrast: While this variable is positively correlated with performance in both the individual-level analysis and the aggregated data analysis without the correction for common-method bias, it is not statistically associated with performance in the aggregated data analysis with the correction. Originality/value – The analysis is the first to assess the robustness of the performance-PSM linkage to a correction for common-method bias. The findings place the validity of at least one part of the individual-level linkage between PSM and performance into question.

Inter-observer agreement for diagnostic classification of esophageal motility disorders defined in high-resolution manometry

High-resolution esophageal manometry (HRM) is a recent development used in the evaluation of esophageal function. Our aim was to assess the inter-observer agreement for diagnosis of esophageal motility disorders using this technology. Practitioners registered on the HRM Working Group website were invited to review and classify (i) 147 individual water swallows and (ii) 40 diagnostic studies comprising 10 swallows using a drop-down menu that followed the Chicago Classification system. Data were presented using a standardized format with pressure contours without a summary of HRM metrics. The sequence of swallows was fixed for each user but randomized between users to avoid sequence bias. Participants were blinded to other entries. (i) Individual swallows were assessed by 18 practitioners (13 institutions). Consensus agreement (≤2/18 dissenters) was present for most cases of normal peristalsis and achalasia but not for cases of peristaltic dysmotility. (ii) Diagnostic studies were assessed by 36 practitioners (28 institutions). Overall inter-observer agreement was 'moderate' (kappa 0.51) being 'substantial' (kappa > 0.7) for achalasia type I/II and no lower than 'fair-moderate' (kappa >0.34) for any diagnosis. Overall agreement was somewhat higher among those that had performed >400 studies (n = 9; kappa 0.55) and 'substantial' among experts involved in development of the Chicago Classification system (n = 4; kappa 0.66). This prospective, randomized, and blinded study reports an acceptable level of inter-observer agreement for HRM diagnoses across the full spectrum of esophageal motility disorders for a large group of clinicians working in a range of medical institutions. Suboptimal agreement for diagnosis of peristaltic motility disorders highlights contribution of objective HRM metrics.

Heme is an effector molecule for iron-dependent degradation of the bacterial iron response regulator (Irr) protein

The bacterial iron response regulator (Irr) protein mediates iron-dependent regulation of heme biosynthesis. Pulse–chase and immunoprecipitation experiments showed that Irr degraded in response to 6 μM iron with a half-life of ≈30 min and that this regulated stability was the principal determinant of control by iron. Irr contains a heme regulatory motif (HRM) near its amino terminus. A role for heme in regulation was implicated by the retention of Irr in heme synthesis mutants in the presence of iron. Addition of heme to low iron (0.3 μM) cultures was sufficient for the disappearance of Irr in cells of the wild-type and heme mutant strains. Spectral and binding analyses of purified recombinant Irr showed that the protein bound heme with high affinity and caused a blue shift in the absorption spectrum of heme to a shorter wavelength. A Cys29 → Ala substitution within the HRM of Irr (IrrC29A) abrogated both high affinity binding to heme and the spectral blue shift. In vivo turnover experiments showed that, unlike wild-type Irr, IrrC29A was stable in the presence of iron. We conclude that iron-dependent degradation of Irr involves direct binding of heme to the protein at the HRM. The findings implicate a regulatory role for heme in protein degradation and provide direct evidence for a functional HRM in a prokaryote.

Down-regulation of dendritic spine and glutamic acid decarboxylase 67 expressions in the reelin haploinsufficient heterozygous reeler mouse

Heterozygous reeler mice (HRM) haploinsufficient for reelin express ≈50% of the brain reelin content of wild-type mice, but are phenotypically different from both wild-type mice and homozygous reeler mice. They exhibit, (i) a down-regulation of glutamic acid decarboxylase 67 (GAD67)-positive neurons in some but not every cortical layer of frontoparietal cortex (FPC), (ii) an increase of neuronal packing density and a decrease of cortical thickness because of neuropil hypoplasia, (iii) a decrease of dendritic spine expression density on basal and apical dendritic branches of motor FPC layer III pyramidal neurons, and (iv) a similar decrease in dendritic spines expressed on the basal dendrite branches of CA1 pyramidal neurons of the hippocampus. To establish whether the defect of GAD67 down-regulation observed in HRM is responsible for neuropil hypoplasia and decreased dendritic spine density, we studied heterozygous GAD67 knockout mice (HG67M). These mice exhibited a down-regulation of GAD67 mRNA expression in FPC (about 50%), but they expressed normal amounts of reelin and had no neuropil hypoplasia or down-regulation of dendritic spine expression. These findings, coupled with electron-microscopic observations that reelin colocalizes with integrin receptors on dendritic spines, suggest that reelin may be a factor in the dynamic expression of cortical dendritic spines perhaps by promoting integrin receptor clustering. These findings are interesting because the brain neurochemical and neuroanatomical phenotypic traits exhibited by the HRM are in several ways similar to those found in postmortem brains of psychotic patients.

A relação entre o ambiente institucional, a gestão de recursos humanos e os resultados organizacionais

Esta tese tem como objetivo verificar as similaridades e diferenças nas práticas de Gestão De Recursos Humanos (GRH) adotadas em empresas inseridas em diferentes tipos de Ambientes Institucionais e suas relações com a percepção dos Resultados Organizacionais. Para isso, adotou-se como premissas: (i) as forças institucionais advindas do Ambiente Institucional brasileiro fazem com que as práticas de GRH das empresas que operam neste contexto tendam a convergir apresentando similaridades e que, concomitantemente, (ii) as forças oriundas das pressões do Ambiente Institucional dos países de origem das empresas fazem com que as práticas de GRH tendam a divergir das práticas locais, apresentando diferenças. Buscou-se embasamento em três teorias: a teorias da Variedades de Capitalismo; a Teoria do Sistema Nacional de Negócios e a tipologia de Economia de Mercado Hierárquico operante na América Latina, Brasil. De natureza positivista, este estudo descritivo e de abordagem contextualista contou com uma população de aproximadamente 22.052 organizações e amostra de 326 empresas. Realizou-se uma survey e fez-se uso de técnicas descritivas e de modelagem de equações estruturais no tratamento e na análise dos dados coletados. Constatou-se que, em geral, há mais similaridades do que diferenças nas práticas de GRH das empresas que operam no Brasil, indo ao encontro da premissa (i). Validou-se que o Ambiente Institucional afeta a GRH no Brasil e esta influencia na percepção dos Resultados Organizacionais das empresas, tanto as com sedes corporativas localizadas no Brasil quanto as com sede em países com economia de mercado liberal. Entretanto, a GRH não demonstrou ser influente nas organizações com sede corporativa localizadas em países de economia coordenada. Comprovou-se que a GRH faz a mediação da relação entre o Ambiente Institucional e os Resultados Organizacionais. Como contribuições, aos práticos destaca-se a reflexão sobre a forma como a GRH concebe suas práticas, quais os limites do papel da área e seu favorecimento à competitividade das empresas. Para a Rede Cranet e acadêmicos, somam-se as evidências sobre o Brasil e seu Ambiente Institucional, ainda pouco explorado. Acredita-se, também, que as análises contribuem para a tomada de decisões dos líderes empresariais ao envolver a relação ambiente e empresa. A contribuição social desta tese situa-se na possibilidade de seu uso em discussões sobre Políticas Públicas referentes a emprego, trabalho, renda, economia e educação.

A gestão de recursos humanos e o desempenho individual

O presente estudo analisou a relação entre a percepção do sistema de práticas de recursos humanos e a percepção de desempenho individual, bem como o papel do compromisso afectivo, enquanto variável mediadora desta relação. Mais especificamente, tendo como referência o modelo de gestão de recursos humanos de Guest, avaliou-se se a percepção que os trabalhadores tinham do sistema de práticas de recursos humanos estava associada à sua percepção de desempenho individual, bem como se o compromisso afectivo, enquanto ligação sócio-emocional que os trabalhadores estabelecem com a sua organização, poderia desenvolver-se como resposta afectiva à percepção do sistema de gestão de recursos humanos e, deste modo, contribuir para explicar a percepção do desempenho individual. As hipóteses foram testadas com modelos de equações estruturais numa amostra de 1885 trabalhadores de uma empresa do sector da banca que responderam a um questionário on-line. Os resultados demonstraram que, tal como esperado, a percepção do sistema de gestão de recursos humanos estava positivamente associada à percepção de desempenho individual e que esta relação era parcialmente mediada pelo compromisso afectivo. No seu conjunto, os resultados deste estudo apontam alguns dos benefícios de uma gestão estratégica de recursos humanos centrada no compromisso dos trabalhadores, sendo discutidas diversas implicações para a gestão de recursos humanos.

A Multiplex Real-Time PCR with High Resolution Melting Analysis for the Characterization of Antimicrobial Resistance in Neisseria gonorrhoeae.

Resistance to antibiotics used against Neisseria gonorrhoeae infections is a major public health concern. Antimicrobial resistance (AMR) testing relies on time-consuming culture-based methods. Development of rapid molecular tests for detecting AMR determinants could provide valuable tools for surveillance, epidemiological studies and to inform individual case management. We developed a fast (<1.5 hrs) SYBR-green based real-time PCR method with high resolution melting (HRM) analysis. One triplex and three duplex reactions included two sequences for N. gonorrhoeae identification and seven determinants of resistance to extended-spectrum cephalosporins (ESCs), azithromycin, ciprofloxacin, and spectinomycin. The method was validated by testing 39 previously fully-characterized N. gonorrhoeae strains, 19 commensal Neisseria spp., and an additional panel of 193 gonococcal isolates. Results were compared with culture-based AMR determination. The assay correctly identified N. gonorrhoeae and the presence or absence of the seven AMR determinants. There was some cross-reactivity with non-gonococcal Neisseria species and the detection limit was 10(3)-10(4) gDNA copies/reaction. Overall, the platform accurately detected resistance to ciprofloxacin (sensitivity and specificity, 100%), ceftriaxone (sensitivity 100%, specificity 90%), cefixime (sensitivity 92%, specificity 94%), azithromycin and spectinomycin (both sensitivity and specificity, 100%). In conclusion, our methodology accurately detects mutations generating resistance to antibiotics used to treat gonorrhea. Low assay sensitivity prevents direct diagnostic testing of clinical specimens but this method can be used to screen collections of gonococcal isolates for AMR more quickly than with current culture-based AMR testing.

Att lansera en artist : En studie av kommunikationskanalerna i den digitala musikvärlden

Syftet med denna uppsats har varit att undersöka hur kommunikationskanaler används av konsumenter i samband med musik, för att lyssna samt att hitta ny musik och artister. Detta för att i förlängningen kunna utreda och förstå vilka av dessa kommunikationskanaler som passar bäst för att lansera artister och göra deras musik tillgänglig på marknaden.   Vi har tagit hjälp av tidigare forskning och teorier om musikindustrins utveckling kopplat till digitaliseringen och uppkomsten av de nya kommunikationskanalerna för att få en bredare bild samt kunna koppla dessa till den kvantitativa enkätundersökning vi genomfört. Teorier kring gräsrotsdriven kommunikation, Word of Mouth och digitalisering har exempelvis varit centrala. Varumärke samt strategisk kommunikation finns även med som begrepp.   Själva metodramen är som sagt av kvantitativ form, där en enkätundersökning ligger till grund för datainsamlingen. Denna har sedan brutits ner och analyserats för att få insikt om respondenternas åsikter och beteende när de konsumerar musik, utifrån vilka kanaler de använder för att lyssna på musik, hitta ny musik samt eventuellt dela och sprida den till vänner och bekanta. Det har även ställts frågor kring vikten av varumärke samt eventuella icke-musikaliska attribut i dagens musikvärld, ifall de spelar in för konsumenten när ny musik uppsöks.   Undersökningen visar att det finns ett tydligt samband kring hur respondenterna dels lyssnar, dels hittar ny musik. Det är genom de nya digitala kommunikationskanalerna detta sker i störst utsträckning. Det visade sig även att det tycks finnas ett dalande förtroende gentemot klassisk uttalad marknadsföring och dess kommunikationskanaler. En majoritet av respondenterna i undersökningen föredrog gräsrotsdrivna kanaler samt mer personliga tips på musik snarare än ren kommersiell marknadsföring. Det var även vanligt att attribut utöver det musikaliska sågs som något positivt, ett mervärde till musiken. När det gäller varumärke kopplat till artister ansåg en majoritet att artister idag faktiskt är att betrakta som ett varumärke.