RESUMO
Cold environments are more frequent than people think. They include deep oceans, cold lakes, snow, permafrost, sea ice, glaciers, cold soils, cold deserts, caves, areas at elevations greater than 3000 m, and also artificial refrigeration systems. These environments are inhabited by a diversity of eukaryotic and prokaryotic organisms that must adapt to the hard conditions imposed by cold. This adaptation is multifactorial and includes (i) sensing the cold, mainly through the modification of the liquid-crystalline membrane state, leading to the activation of a two-component system that transduce the signal; (ii) adapting the composition of membranes for proper functions mainly due to the production of double bonds in lipids, changes in hopanoid composition, and the inclusion of pigments; (iii) producing cold-adapted proteins, some of which show modifications in the composition of amino acids involved in stabilizing interactions and structural adaptations, e.g., enzymes with high catalytic efficiency; and (iv) producing ice-binding proteins and anti-freeze proteins, extracellular polysaccharides and compatible solutes that protect cells from intracellular and extracellular ice. However, organisms also respond by reprogramming their metabolism and specifically inducing cold-shock and cold-adaptation genes through strategies such as DNA supercoiling, distinctive signatures in promoter regions and/or the action of CSPs on mRNAs, among others. In this review, we describe the main findings about how organisms adapt to cold, with a focus in prokaryotes and linking the information with findings in eukaryotes.
Assuntos
Adaptação Fisiológica , Proteínas , Humanos , Adaptação Fisiológica/fisiologia , Proteínas/metabolismo , Aminoácidos , Oceanos e Mares , Solo , Temperatura BaixaRESUMO
Nuclear receptors are ligand-activated transcription factors capable of regulating the expression of complex gene networks. The family includes seven subfamilies of protein with a wide phylogenetic distribution. A novel subfamily with two DNA-binding domains (2DBDs) has been first reported in Schistosoma mansoni (Platyhelminth, Trematoda). Employing an ab initio protocol and homology modeling methods, the full-length 3D structure of the Eg2DBDα.1 nuclear receptor from Echinococcus granulosus (Platyhelminth, Cestoda) was generated. The model analysis reveals the presence of the conserved three-layered alpha-helical sandwich structure in the ligand binding domain, and a particularly long and flexible hinge region. Molecular dynamics simulations were performed previous to dock a conformational library of fatty acids and retinoic acids. Our results indicate that oleic and linoleic acids are suitable ligands to this receptor. The ligand-protein complex is stabilized mainly by hydrogen bonds and hydrophobic interactions. The fact that 2DBD nuclear receptors have not been identified in vertebrates confers particular interest to these nuclear receptors, not only concerning their structure and function but as targets of new anthelmintic drugs.Communicated by Ramaswamy H. Sarma.
Assuntos
Echinococcus granulosus , Animais , Echinococcus granulosus/metabolismo , Simulação de Dinâmica Molecular , Filogenia , Ligantes , Proteínas de Helminto/química , Receptores Citoplasmáticos e Nucleares , Simulação de Acoplamento MolecularRESUMO
The intestinal fatty acid binding protein (FABP) is a small protein expressed along the small intestine that bind long-chain fatty acids and other hydrophobic ligands. Several lines of evidence suggest that, once in the nucleus, it interacts with nuclear receptors, activating them and thus transferring the bound ligand into the nucleus. Previous work by our group suggests that FABP2 would participate in the cytoplasm-nucleus translocation of fatty acids. Because the consensus NLS is absent in the sequence of FABP2, we propose that a 3D signal could be responsible for its nuclear translocation. The results obtained by transfection assays of recombinant wild type and mutated forms of Danio rerio Fabp2 in Caco-2 cell cultures, showed that lysine 17, arginine 29 and lysine 30 residues, which are located in the helix-turn-helix region, would constitute a functional non-classical three-dimensional NLS.
Assuntos
Núcleo Celular/metabolismo , Proteínas de Ligação a Ácido Graxo/metabolismo , Sequência de Aminoácidos , Animais , Células CACO-2 , Proteínas de Ligação a Ácido Graxo/química , Proteínas de Ligação a Ácido Graxo/genética , Ácidos Graxos/metabolismo , Humanos , Microscopia Confocal , Mutagênese , Sinais de Localização Nuclear/química , Ligação Proteica , Conformação Proteica em alfa-Hélice , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/química , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoRESUMO
Nuclear receptors are ligand-activated transcription factors capable of regulating the expression of complex gene networks. The family includes seven subfamilies of proteins with a wide phylogenetic distribution. A novel subfamily with two DNA-binding domains (2DBDs) has been reported in Schistosoma mansoni (Platyhelminth, Trematoda). This work describes the cDNA cloning and bioinformatics analysis of Eg2DBDα, a 2DBD nuclear receptor isoform from the parasite Echinococcus granulosus (Platyhelminth, Cestoda). The Eg2DBDα gene coding domain structure was analysed. Although two additional 2DBD nuclear receptors are reported in the parasite database GeneDB, they are unlikely to be expressed in the larval stage. Phylogenetic relationships between these atypical proteins from different cestodes are also analysed including S. mansoni 2DBD nuclear receptors. The presence of two DNA binding domains confers particular interest to these nuclear receptors, not only concerning their function but to the development of new antihelminthic drugs.
Assuntos
Echinococcus granulosus/metabolismo , Proteínas de Helminto/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Animais , Biologia Computacional , Echinococcus granulosus/genética , Proteínas de Helminto/genética , Filogenia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Receptores Citoplasmáticos e Nucleares/genéticaRESUMO
Introdução: Esta comunicação compartilha a experiência de um trabalho interinstitucional, envolvendo atividades de promoção da saúde e prevenção de agravos à saúde vocal de professores da rede municipal de São Paulo, por meio de educação em saúde na modalidade de Ensino à Distância. Método: Participaram do processo estudantes, docentes da Pontifícia Universidade Católica de São Paulo e coordenadora do Programa Municipal de Saúde Vocal. A oficina continha 40 horas, com dois encontros presenciais e os demais on-line: 8 módulos, 8 Fóruns, 8 avaliações e 16 chats. 75 professores participaram da proposta e responderam um questionário aplicado ao final para avaliar forma e conteúdo da oficina. Resultados: A experiência mostrou-se positiva, tanto para o público-alvo (professores), como na formação dos profissionais (fonoaudiólogos) e gestores com responsabilidade de atender necessidades de saúde da população. A maioria dos professores informou que a oficina foi importante para o aprendizado profissional (94%); fez refletir sobre o conteúdo (94%); possibilitou expor ideias a outros participantes (68%); teve apoio de tutores (81%); sentiu-se encorajado a permanecer no curso (80%); e acredita que o curso teve tempo de duração ideal (77%). Conclusão: A integração entre universidade e serviços possibilita o desenvolvimento de experiências que vão ao encontro das necessidades de saúde da população. Ações de educação em saúde na modalidade à distância podem ser uma opção importante para o trabalho com maior número de pessoas, visando à promoção de saúde e prevenção de agravos e o autocuidado de professores.
Introduction: This paper shares the experience of interinstitutional work, involving health promotion activities and disease prevention to vocal health of teachers from public schools of São Paulo city, through health education in distance learning mode. Methods: Participated in the process: students, teachers at Pontifícia Universidade Católica de São Paulo and coordinator of the Municipal Health Program Vocal of São Paulo city. The workshop contained 40 hours, with two face meetings and others on-line: 8 modules, 8 Forums, 8 reviews and 16 chats. 75 teachers participated in the proposal. At the end, they answered a questionnaire to asses the form and content of workshop. Results: The experience was positive both for the target audience (teachers) and in the training of professionals (speech therapists) and managers with responsibility to serve the health needs of the population. Most teachers reported that the workshop was important for professional learning (94%); did reflect on the content (94%); enabled expose ideas to other participants (68%); we had support from tutors (81%); was encouraged to continue the course (80%); and believes the course was ideal duration (77%). Conclusion: Integration between university and service enables the development of experiences that meet the health needs of the population. Health education actions in distance learning mode can be an important option to work with more people in order to promote health, disease prevention and self-care teachers.
Introducción: Este comunicado comparte la experiencia de un trabajo inter-institucional, envolviendo actividades de promoción de la salud y prevención de agravas a la salud vocal de los profesores de la red municipal de San Pablo, por medio de la educación en salud en la modalidad de Enseñanza a Distancia. Método: participaron del estudio estudiantes, docentes de la Pontificia Universidad Católica de San Pablo y la coordinadora del Programa Municipal de Salud Vocal. El taller se duró de 40 horas, con dos encuentros presenciales y los demás on-line: 8 módulos, 8 Fórums, 8 evaluaciones y 16 chats. Participaron de la propuesta 75 profesores que contestaron un cuestionario ministrado al final para evaluar la forma y el contenido del taller. Resultados: La experiencia se mostró positiva tanto para el públicoobjetivo (profesores) como para la formación de los profesionales (fonoaudiólogos) y gestores responsables por la atención de las necesidades de salud de la población. La mayoría de los profesores informó que el taller fue importante para el aprendizaje profesional (94%); hizo reflexionar sobre el contenido (94%); posibilitó la exposición de ideas a otros participantes (68%); obtuvo apoyo de tutores (81%); se sintió encorajado en permanecer en el curso (80%), cree que el tiempo de duración del curso fue ideal (77%). Conclusión: la integración entre la universidad y los servicios posibilita el desarrollo de experiencias que van al encuentro de las necesidades de salud de la población. Acciones de educación en salud en la modalidad a distancia pueden ser una opción importante para el trabajo con mayor número de personas, visionando la promoción de la salud y prevención de agravas y autocuidado de los profesores.
Assuntos
Humanos , Adulto , Educação a Distância , Docentes , Promoção da Saúde , Colaboração Intersetorial , VozRESUMO
Two fatty acid binding proteins, EgFABP1 and EgFABP2, were isolated from the parasitic platyhelminth Echinococcus granulosus. These proteins bind fatty acids and have particular relevance in flatworms since de novo fatty acids synthesis is absent. Therefore platyhelminthes depend on the capture and intracellular distribution of host's lipids and fatty acid binding proteins could participate in lipid distribution. To elucidate EgFABP's roles, we investigated their intracellular distribution in the larval stage by a proteomic approach. Our results demonstrated the presence of EgFABP1 isoforms in cytosolic, nuclear, mitochondrial and microsomal fractions, suggesting that these molecules could be involved in several cellular processes.
Assuntos
Echinococcus granulosus/química , Proteínas de Ligação a Ácido Graxo/análise , Proteínas de Helminto/análise , Animais , Western Blotting , Bovinos , Doenças dos Bovinos/parasitologia , Equinococose/parasitologia , Equinococose/veterinária , Echinococcus granulosus/metabolismo , Echinococcus granulosus/ultraestrutura , Eletroforese em Gel Bidimensional , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Regulação da Expressão Gênica , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Interações Hospedeiro-Parasita , Larva/química , Larva/metabolismo , Larva/ultraestrutura , Metabolismo dos Lipídeos , Isoformas de Proteínas/análise , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteômica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , UruguaiRESUMO
Intracellular lipid binding proteins, including fatty acid binding proteins (FABPs) 1 and 2, are highly expressed in tissues involved in the active lipid metabolism. A zebrafish model was used to demonstrate differential expression levels of fabp1b.1, fabp1b.2, and fabp2 transcripts in liver, anterior intestine, and brain. Transcription levels of fabp1b.1 and fabp2 in the anterior intestine were upregulated after feeding and modulated according to diet formulation. Immunofluorescence and electron microscopy immunodetection with gold particles localized these FABPs in the microvilli, cytosol, and nuclei of most enterocytes in the anterior intestinal mucosa. Nuclear localization was mostly in the interchromatin space outside the condensed chromatin clusters. Native PAGE binding assay of BODIPY-FL-labeled FAs demonstrated binding of BODIPY-FLC(12) but not BODIPY-FLC(5) to recombinant Fabp1b.1 and Fabp2. The binding of BODIPY-FLC(12) to Fabp1b.1 was fully displaced by oleic acid. In vivo experiments demonstrated, for the first time, that intestinal absorption of dietary BODIPY-FLC(12) was followed by colocalization of the labeled FA with Fabp1b and Fabp2 in the nuclei. These data suggest that dietary FAs complexed with FABPs are able to reach the enterocyte nucleus with the potential to modulate nuclear activity.
Assuntos
Proteínas de Ligação a Ácido Graxo/metabolismo , Ácidos Graxos/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Peixe-Zebra/genética , Animais , Núcleo Celular/genética , Núcleo Celular/metabolismo , Citosol/metabolismo , Enterócitos/metabolismo , Proteínas de Ligação a Ácido Graxo/genética , Regulação da Expressão Gênica , Humanos , Mucosa Intestinal/metabolismo , Metabolismo dos Lipídeos/genética , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genéticaRESUMO
Two fatty acid binding proteins, MvFABPa and MvFABPb were identified in the parasite Mesocestoides vogae (Platyhelmithes, Cestoda). Fatty acid binding proteins are small intracellular proteins whose members exhibit great diversity. Proteins of this family have been identified in many organisms, of which Platyhelminthes are among the most primitive. These proteins have particular relevance in flatworms since de novo synthesis of fatty acids is absent. Fatty acids should be captured from the media needing an efficient transport system to uptake and distribute these molecules. While HLBPs could be involved in the shuttle of fatty acids to the surrounding host tissues and convey them into the parasite, FABPs could be responsible for the intracellular trafficking. In an effort to understand the role of MvFABPs in fatty acid transport of M. vogae larvae, we analysed the intracellular localization of both MvFABPs and the co-localization with in vivo uptake of fatty acid analogue BODIPY FL C16. Immunohistochemical studies on larvae sections using specific antibodies, showed a diffuse cytoplasmic distribution of each protein with some expression in nuclei and mitochondria. MvFABPs distribution was confirmed by mass spectrometry identification from 2D-electrophoresis of larvae subcellular fractions. This work is the first report showing intracellular distribution of MvFABPs as well as the co-localization of these proteins with the BODIPY FL C16 incorporated from the media. Our results suggest that fatty acid binding proteins could target fatty acids to cellular compartments including nuclei. In this sense, M. vogae FABPs could participate in several cellular processes fulfilling most of the functions attributed to vertebrate's counterparts.
Assuntos
Proteínas de Ligação a Ácido Graxo/metabolismo , Proteínas de Helminto/metabolismo , Mesocestoides/metabolismo , Sequência de Aminoácidos , Animais , Proteínas de Ligação a Ácido Graxo/química , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Helminto/química , Proteínas de Helminto/genética , Dados de Sequência Molecular , Transporte ProteicoRESUMO
Fatty acid (FA) binding proteins are small intracellular proteins whose members exhibit great diversity and low similarity at the primary structure level, but a highly conserved three-dimensional structure. Characterised by a high-affinity non-covalent binding of hydrophobic ligands, these proteins have a molecular mass of 14-15 kDa with a characteristic ß-barrel structure. Members of this family have been identified along the zoological scale, with Platyhelminthes being the more primitive organisms where they have been reported. Two FA binding proteins (FABPs), EgFABP1 and EgFABP2, with 88% similarity have been identified in Echinococcus granulosus. In an effort to understand why two such similar proteins are expressed by this organism, we performed an in silico analysis of the binding capabilities of both proteins. The crystallographic structure of EgFABP1 was utilised as a template to model EgFABP2, and both were docked against palmitate, oleate, linoleate and arachidonate. The docked structures were submitted to 4 ns molecular dynamics simulations, and their protein-ligand interaction energies were measured. The collected data demonstrated that linoleate and arachidonate had the higher interaction energies when bound to EgFABP1 and that palmitate and linoleate had the higher interaction energies when bound to EgFABP2. External and internal binding surfaces were analysed, showing differences at both levels. Internal surface compositions suggested that both proteins could have preferences for certain FAs. Comparisons of the holo and apo forms of each protein indicated that the ligand imposed subtle, but specific modifications that could trigger surface signals. The differences found between the proteins under study suggest that they could have functional uniqueness in the parasite's metabolism.
Assuntos
Echinococcus granulosus , Proteínas de Ligação a Ácido Graxo/química , Proteínas de Helminto/química , Simulação de Dinâmica Molecular , Sequência de Aminoácidos , Animais , Apoproteínas/química , Ácido Araquidônico/química , Sítios de Ligação , Ligação de Hidrogênio , Ligantes , Simulação de Acoplamento Molecular , Dados de Sequência Molecular , Ácidos Oleicos/química , Palmitatos/química , Ligação Proteica , Estrutura Secundária de Proteína , Homologia Estrutural de Proteína , Termodinâmica , Água/químicaRESUMO
Introdução: Muitos estudos, no campo da Fonoaudiologia, têm se preocupado em estudar a voz dos professores, a fim de pensar ações de promoção de saúde e/ou prevenção de alterações vocais. Objetivo:levantar o conhecimento dos professores acerca dos fatores predisponentes para o bem-estar vocal, além de verificar o uso de práticas que favorecem o bem-estar vocal, segundo o tempo de docência.Material e Método: A amostra deste estudo, de tipo transversal, foi constituída por 242 professores da rede pública do município de Jataí-GO, que antes de uma oficina educativa assistiram ao vídeo ?O que é bom para o dono é bom para a voz? e preencheram um instrumento com seis questões relacionadas ao bem-estar vocal, hábitos e exercícios vocais. Foram consideradas as respostas de duas questões (informações novas e hábitos praticados) tabuladas e analisadas (qui-quadrado), quanto ao tempo de docência. Resultados: Foram consideradas informações novas e não realizadas pelos professores as que faziam referência a exercícios de aquecimento e desaquecimento vocal; entre os de conhecimentoe praticado pelos professores destacaram-se hábitos que interferem na voz (falar alto, falar muito, tossir). Não foi registrada diferença na associação entre o tempo de profissão e a maioria dos aspectos(exceção a prática de ?bocejar?) Conclusão: As respostas indicaram que mesmo tendo conhecimento de fatores prejudiciais ao bem-estar vocal, os mesmos permanecem fazendo parte da rotina e da prática dos professores, fato que alerta o fonoaudiólogo na condução de suas ações junto a esse profissional...
Introduction: In the fi eld of Speech-Language Pathology, many studies have been focusing on the voice of teachers, in order to propose actions in health promotion and/or prevention of voice disorders. Aim: Raise teachers? knowledge about risk factors for the vocal well-being, and check the use of practices that favor the vocal well-being, according to the number of years of teaching. Material and Method: The sample of this cross-sectional study was composed by 242 teachers of the public school system of the city of Jataí-GO, who, before an educational workshop watched a specific video about teachers? voice, and filled out an instrument composed of six questions about vocal health, habits and vocal exercises.The answers to two questions (new information and habits) were considered and analyzed (chi-squaretest), according to years of teaching. Results: The considered data regarded new information not putinto practice by the teachers, the information that concerned vocal warm-up and cool-down. Among theinformation known and practiced by the teachers deleterious habits to the voice that were used (speckingloudly, speaking excessively, coughing) were highlighted. There was no difference in the association ofyears of teaching and most aspects (with exception of ?yawning?). Conclusion: The answers indicated thateven knowing about deleterious factors to vocal well-being, these continue to be a part of the teachers?routine and habits. This fact should alert Speech-Language Pathologists in conducting their practiceswith this professional category...
Introducción: Muchos estudios en el campo de la Logopedia, se han ocupado de estudiar la voz de los docentes, con el fi n de refl ejar las acciones de promoción de la salud y/ o prevención de trastornosde la voz. Objetivo: investigar el conocimiento de los profesores acerca de los factores predisponentes para el bienestar vocal, y verificar el uso de prácticas que favorezcan el bienestar vocal, en función del tiempo de docencia. Material y Método: La muestra para este estudio transversal se componía por 242 maestros de escuela pública en el municipio de Jataí-GO. Antes de un taller educativo asistieron el video ?O que é bom para o dono é bom para a voz? y respondieron un cuestionario con seis temas de la salud vocal, hábitos y ejercicios vocales. Se consideró dos preguntas (nuevas informaciones y los hábitos de práctica) y estos fueron tabulados e analizados (qui-quadrado), cuanto el tiempo de la enseñanza. Resultados: Se consideró que las nuevas informaciones y las no practicadas por los maestros que hacían referencia a los ejercicios de calentamiento y enfriamiento vocal; los aspectos conocidos e practicadospor los profesionales fueron los hábitos que interfi eren en la voz (hablar fuerte y en exceso, mucho. No hubo diferencias en la asociación del tiempo de la profesión y la mayoría de los aspectos (excepto en la práctica del ?bostezo?). Conclusión: Las respuestas indicaron que aún con conocimiento de los factores perjudiciales para el bienestar vocal, siguen siendo parte de la rutina e de la práctica de estos profesionales, esto alerta el logopedista para pensar las acciones con estos profesionales...
Assuntos
Humanos , Docentes , Fatores de Risco , VozRESUMO
Objetivo: avaliar a reprodutibilidade teste-reteste das questões referentes ao levantamento de sintomas vocais e sensações laringo-faríngeas, presentes no instrumento Condições de Produção Vocal do Professor (CPV-P). Método: Cem professoras, que compareceram a um hospital público de São Paulo com queixa de alteração vocal, responderam ao questionário CPV-P. Dessas, 36 responderam o mesmo instrumento com intervalo de 15 dias entre as aplicações. As questões sociodemográficas foram utilizadas para a caracterização da amostra e as variáveis referentes aos sintomas vocais e sensações laringofaríngeas foram analisadas de forma descritiva e comparadas quanto à situação teste-reteste, por meio da estatística Kappa (k) (p ? 0,05), a fi m de avaliar a concordância entre as duas aplicações do instrumento. Resultados: A concordância bruta dos sintomas vocais (rouquidão, falha na voz, voz grossa e voz fraca) e sensações laringofaríngeas (bolo na garganta, pigarro, dor ao falar, ardor na garganta, garganta seca, cansaço ao falar e esforço ao falar) analisados foi superior a 75% e os valores de kpara os sintomas vocais variaram de 0,52 a 0,74 e para as sensações laringo-faríngeas de 0,36 a 0,59. Conclusão: Os resultados indicaram no estudo de reprodutibilidade teste-reteste, nível de concordância de regular a forte...
Aim: to evaluate the test-retest reproducibility of the questions pertaining to the list of vocal symptoms and laryngeal-pharyngeal sensations present in the instrument named Vocal Production Conditions of Teachers (VPC-T). Method: One hundred teachers who sought attention at a public hospital in São Paulo with voice complaints responded to the VPC-T questionnaire. Of these, 36 completed the same instrument after an interval of 15 days. The social demographic questions were used in order to characterize thesample and the variables regarding vocal symptoms and laryngeal pharyngeal sensations were analyzed descriptively and compared for test-retest situations, using Kappa (k) statistics (p?0.05) in order to assessthe agreement between both questionnaire completions. Results: The gross agreement of the analyzed vocal symptoms (hoarseness, breaking voice, low-pitched voice and weak voice) and of laryngeal pharyngeal sensations (globus, phlegm, pain when speaking, sore throat, dry throat and strained speech) was higher than 75% and the k values for vocal symptoms varied in between 0,52 and 0,74 and for laryngeal pharyngeal sensations in between 0,36 and 0,59. Conclusion: The results showed agreement varying from regular to strong in the study for test-retest reproducibility...
Introducción: reproducibilidad test-retest es una medida que evalúa la estabilidad de un instrumento, por lo que los procedimientos de prueba de conducción y también en lo que respecta a las características evaluadas. Objetivo: Evaluar la reproducibilidad test-retest de las cuestiones relativas a la lista de los síntomas vocales y la laringe o de la faringe sensaciones presentes en el instrumento llamado condiciones vocales de producción de los maestros (CPV-M). Método: Un centenar de maestros que buscaban atención en un hospital público de São Paulo, con las quejas de voz respondieron al cuestionario CPV-M. De éstos, 36 completaron el mismo instrumento después de un intervalo de 15 días. Las preguntas demográficas sociales fueron utilizados para caracterizar la muestra y las variables relacionadas con los síntomas vocales y laríngeos sensaciones faríngeas se realizó un análisis descriptivo y comparado de las situacionesde test-retest, con Kappa (k) (p ? 0,05) con el fin de evaluar el acuerdo entre las terminaciones de los dos cuestionarios. Resultados: El acuerdo bruto de los síntomas analizados vocales (ronquera, voz entrecortada, la voz de tono bajo y la voz débil) y de la laringe sensaciones faríngeas (globo, flema, dolor al hablar, dolor de garganta, sequedad en la garganta y la voz tensa) fue mayor que 75% y los valores de k para los síntomas vocales variaron en entre 0,52 y 0,74 y para laríngeos sensaciones faríngeas en entre 0,36 y 0,59. Conclusión: Los resultados mostraron que varía de acuerdo de regular a fuerte en el estudio de la reproducibilidad test-retest...
Assuntos
Humanos , Adulto , Docentes , Reprodutibilidade dos Testes , Distúrbios da VozRESUMO
Objetivo: avaliar a reprodutibilidade teste-reteste das questões referentes ao levantamento de sintomas vocais e sensações laringo-faríngeas, presentes no instrumento Condições de Produção Vocal do Professor (CPV-P). Método: Cem professoras, que compareceram a um hospital público de São Paulo com queixa de alteração vocal, responderam ao questionário CPV-P. Dessas, 36 responderam o mesmo instrumento com intervalo de 15 dias entre as aplicações. As questões sociodemográficas foram utilizadas para a caracterização da amostra e as variáveis referentes aos sintomas vocais e sensações laringofaríngeas foram analisadas de forma descritiva e comparadas quanto à situação teste-reteste, por meio da estatística Kappa (k) (p ? 0,05), a fi m de avaliar a concordância entre as duas aplicações do instrumento. Resultados: A concordância bruta dos sintomas vocais (rouquidão, falha na voz, voz grossa e voz fraca) e sensações laringofaríngeas (bolo na garganta, pigarro, dor ao falar, ardor na garganta, garganta seca, cansaço ao falar e esforço ao falar) analisados foi superior a 75% e os valores de kpara os sintomas vocais variaram de 0,52 a 0,74 e para as sensações laringo-faríngeas de 0,36 a 0,59. Conclusão: Os resultados indicaram no estudo de reprodutibilidade teste-reteste, nível de concordância de regular a forte.(AU)
Aim: to evaluate the test-retest reproducibility of the questions pertaining to the list of vocal symptoms and laryngeal-pharyngeal sensations present in the instrument named Vocal Production Conditions of Teachers (VPC-T). Method: One hundred teachers who sought attention at a public hospital in São Paulo with voice complaints responded to the VPC-T questionnaire. Of these, 36 completed the same instrument after an interval of 15 days. The social demographic questions were used in order to characterize thesample and the variables regarding vocal symptoms and laryngeal pharyngeal sensations were analyzed descriptively and compared for test-retest situations, using Kappa (k) statistics (p?0.05) in order to assessthe agreement between both questionnaire completions. Results: The gross agreement of the analyzed vocal symptoms (hoarseness, breaking voice, low-pitched voice and weak voice) and of laryngeal pharyngeal sensations (globus, phlegm, pain when speaking, sore throat, dry throat and strained speech) was higher than 75% and the k values for vocal symptoms varied in between 0,52 and 0,74 and for laryngeal pharyngeal sensations in between 0,36 and 0,59. Conclusion: The results showed agreement varying from regular to strong in the study for test-retest reproducibility.(AU)
Introducción: reproducibilidad test-retest es una medida que evalúa la estabilidad de un instrumento, por lo que los procedimientos de prueba de conducción y también en lo que respecta a las características evaluadas. Objetivo: Evaluar la reproducibilidad test-retest de las cuestiones relativas a la lista de los síntomas vocales y la laringe o de la faringe sensaciones presentes en el instrumento llamado condiciones vocales de producción de los maestros (CPV-M). Método: Un centenar de maestros que buscaban atención en un hospital público de São Paulo, con las quejas de voz respondieron al cuestionario CPV-M. De éstos, 36 completaron el mismo instrumento después de un intervalo de 15 días. Las preguntas demográficas sociales fueron utilizados para caracterizar la muestra y las variables relacionadas con los síntomas vocales y laríngeos sensaciones faríngeas se realizó un análisis descriptivo y comparado de las situacionesde test-retest, con Kappa (k) (p ? 0,05) con el fin de evaluar el acuerdo entre las terminaciones de los dos cuestionarios. Resultados: El acuerdo bruto de los síntomas analizados vocales (ronquera, voz entrecortada, la voz de tono bajo y la voz débil) y de la laringe sensaciones faríngeas (globo, flema, dolor al hablar, dolor de garganta, sequedad en la garganta y la voz tensa) fue mayor que 75% y los valores de k para los síntomas vocales variaron en entre 0,52 y 0,74 y para laríngeos sensaciones faríngeas en entre 0,36 y 0,59. Conclusión: Los resultados mostraron que varía de acuerdo de regular a fuerte en el estudio de la reproducibilidad test-retest.(AU)
Assuntos
Humanos , Adulto , Reprodutibilidade dos Testes , Distúrbios da Voz , DocentesRESUMO
Introdução: Muitos estudos, no campo da Fonoaudiologia, têm se preocupado em estudar a voz dos professores, a fim de pensar ações de promoção de saúde e/ou prevenção de alterações vocais. Objetivo:levantar o conhecimento dos professores acerca dos fatores predisponentes para o bem-estar vocal, além de verificar o uso de práticas que favorecem o bem-estar vocal, segundo o tempo de docência.Material e Método: A amostra deste estudo, de tipo transversal, foi constituída por 242 professores da rede pública do município de Jataí-GO, que antes de uma oficina educativa assistiram ao vídeo ?O que é bom para o dono é bom para a voz? e preencheram um instrumento com seis questões relacionadas ao bem-estar vocal, hábitos e exercícios vocais. Foram consideradas as respostas de duas questões (informações novas e hábitos praticados) tabuladas e analisadas (qui-quadrado), quanto ao tempo de docência. Resultados: Foram consideradas informações novas e não realizadas pelos professores as que faziam referência a exercícios de aquecimento e desaquecimento vocal; entre os de conhecimentoe praticado pelos professores destacaram-se hábitos que interferem na voz (falar alto, falar muito, tossir). Não foi registrada diferença na associação entre o tempo de profissão e a maioria dos aspectos(exceção a prática de ?bocejar?) Conclusão: As respostas indicaram que mesmo tendo conhecimento de fatores prejudiciais ao bem-estar vocal, os mesmos permanecem fazendo parte da rotina e da prática dos professores, fato que alerta o fonoaudiólogo na condução de suas ações junto a esse profissional.(AU)
Introduction: In the fi eld of Speech-Language Pathology, many studies have been focusing on the voice of teachers, in order to propose actions in health promotion and/or prevention of voice disorders. Aim: Raise teachers? knowledge about risk factors for the vocal well-being, and check the use of practices that favor the vocal well-being, according to the number of years of teaching. Material and Method: The sample of this cross-sectional study was composed by 242 teachers of the public school system of the city of Jataí-GO, who, before an educational workshop watched a specific video about teachers? voice, and filled out an instrument composed of six questions about vocal health, habits and vocal exercises.The answers to two questions (new information and habits) were considered and analyzed (chi-squaretest), according to years of teaching. Results: The considered data regarded new information not putinto practice by the teachers, the information that concerned vocal warm-up and cool-down. Among theinformation known and practiced by the teachers deleterious habits to the voice that were used (speckingloudly, speaking excessively, coughing) were highlighted. There was no difference in the association ofyears of teaching and most aspects (with exception of ?yawning?). Conclusion: The answers indicated thateven knowing about deleterious factors to vocal well-being, these continue to be a part of the teachers?routine and habits. This fact should alert Speech-Language Pathologists in conducting their practiceswith this professional category(AU)
Introducción: Muchos estudios en el campo de la Logopedia, se han ocupado de estudiar la voz de los docentes, con el fi n de refl ejar las acciones de promoción de la salud y/ o prevención de trastornosde la voz. Objetivo: investigar el conocimiento de los profesores acerca de los factores predisponentes para el bienestar vocal, y verificar el uso de prácticas que favorezcan el bienestar vocal, en función del tiempo de docencia. Material y Método: La muestra para este estudio transversal se componía por 242 maestros de escuela pública en el municipio de Jataí-GO. Antes de un taller educativo asistieron el video ?O que é bom para o dono é bom para a voz? y respondieron un cuestionario con seis temas de la salud vocal, hábitos y ejercicios vocales. Se consideró dos preguntas (nuevas informaciones y los hábitos de práctica) y estos fueron tabulados e analizados (qui-quadrado), cuanto el tiempo de la enseñanza. Resultados: Se consideró que las nuevas informaciones y las no practicadas por los maestros que hacían referencia a los ejercicios de calentamiento y enfriamiento vocal; los aspectos conocidos e practicadospor los profesionales fueron los hábitos que interfi eren en la voz (hablar fuerte y en exceso, mucho. No hubo diferencias en la asociación del tiempo de la profesión y la mayoría de los aspectos (excepto en la práctica del ?bostezo?). Conclusión: Las respuestas indicaron que aún con conocimiento de los factores perjudiciales para el bienestar vocal, siguen siendo parte de la rutina e de la práctica de estos profesionales, esto alerta el logopedista para pensar las acciones con estos profesionales.(AU)
Assuntos
Humanos , Voz , Docentes , Fatores de RiscoRESUMO
BACKGROUND: Growth and maintenance of hydatid cysts produced by Echinococcus granulosus have a high requirement for host lipids for biosynthetic processes, membrane building and possibly cellular and developmental signalling. This requires a high degree of lipid trafficking facilitated by lipid transporter proteins. Members of the fatty acid binding protein (FABP) family have been identified in Echinococcus granulosus, one of which, EgFABP1 is expressed at the tegumental level in the protoscoleces, but it has also been described in both hydatid cyst fluid and secretions of protoscoleces. In spite of a considerable amount of structural and biophysical information on the FABPs in general, their specific functions remain mysterious. METHODOLOGY/PRINCIPAL FINDINGS: We have investigated the way in which EgFABP1 may interact with membranes using a variety of fluorescence-based techniques and artificial small unilamellar vesicles. We first found that bacterial recombinant EgFABP1 is loaded with fatty acids from the synthesising bacteria, and that fatty acid binding increases its resistance to proteinases, possibly due to subtle conformational changes induced on EgFABP1. By manipulating the composition of lipid vesicles and the ionic environment, we found that EgFABP1 interacts with membranes in a direct contact, collisional, manner to exchange ligand, involving both ionic and hydrophobic interactions. Moreover, we observed that the protein can compete with cytochrome c for association with the surface of small unilamellar vesicles (SUVs). CONCLUSIONS/SIGNIFICANCE: This work constitutes a first approach to the understanding of protein-membrane interactions of EgFABP1. The results suggest that this protein may be actively involved in the exchange and transport of fatty acids between different membranes and cellular compartments within the parasite.
Assuntos
Echinococcus granulosus/fisiologia , Proteínas de Ligação a Ácido Graxo/metabolismo , Membranas/metabolismo , Fosfolipídeos/metabolismo , Animais , Ligação ProteicaRESUMO
TWO MAIN FAMILIES OF LIPID BINDING PROTEINS HAVE BEEN IDENTIFIED IN PARASITIC PLATYHELMINTHES: hydrophobic ligand binding proteins (HLBPs) and fatty acid binding proteins (FABPs). Members of the former family of proteins are specific to the Cestoda class, while FABPs are conserved across a wide range of animal species. Because Platyhelminthes are unable to synthesize their own lipids, these lipid-binding proteins are important molecules in these organisms. HLBPs are a high molecular mass complex of proteins and lipids. They are composed of subunits of low molecular mass proteins and a wide array of lipid molecules ranging from CoA esters to cholesterol. These proteins are excretory-secretory molecules and are key serological tools for diagnosis of diseases caused by cestodes. FABPs are mainly intracellular proteins of low molecular weight. They are also vaccine candidates. Despite that the knowledge of their function is scarce, the differences in their molecular organization, ligand preferences, intra/extracellular localization, evolution, and phylogenetic distribution, suggest that platyhelminths HLBPs and FABPs should play different functions. FABPs might be involved in the removal of fatty acids from the inner surface of the cell membrane and in their subsequent targeting to specific cellular destinations. In contrast, HLBPs might be involved in fatty acid uptake from the host environment.
RESUMO
Tropomyosins (Trps) constitute a family of actin filament-binding proteins found in all eukaryotic cells. In muscle cells, they play a central role in contraction by regulating calcium-sensitive interaction of actin and myosin. In non-muscle cells, tropomyosins regulate actin filament organization and dynamics. Trps genes exhibit extensive cell type-specific isoform diversity generated by alternative splicing. Here, we report the characterization of tropomyosin gene transcribed sequences from the parasitic platyhelminth Echinococcus granulosus. Using RT-PCR approach we isolated three isoforms (egtrpA, egtrpB and egtrpC), which display significant homologies to know tropomyosins of different phylogenetic origin. The corresponding gene, egtrp (5656 bp), contains eight introns and nine exons. Southern blot hybridization studies showed that egtrp is present as single copy locus in E. granulosus. We demonstrated that egtrp expresses three different transcripts which differ in alternatively spliced exon 4 and intron VI. Interestingly, intron VI suffers intron retention and contains an internal stop codon in frame. Three major bands are also detected by Western blot analysis using a specific anti-rEgTrp antiserum. Immune-localization and in situ hybridization studies showed that egtrp transcription and translation is mostly localized at the protoscoleces suckers. This is the first report of alternative splicing in this parasite.
Assuntos
Echinococcus granulosus/genética , Expressão Gênica , Isoformas de Proteínas/genética , Tropomiosina/genética , Processamento Alternativo , Animais , Sequência de Bases , Southern Blotting , Western Blotting , Primers do DNA , Éxons , Hibridização In Situ , Íntrons , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
This work describes two new fatty acid binding proteins (FABPs) identified in the parasite platyhelminth Mesocestoides vogae (syn. corti). The corresponding polypeptide chains share 62% identical residues and overall 90% similarity according to CLUSTALX default conditions. Compared with Cestoda FABPs, these proteins share the highest similarity score with the Taenia solium protein. M. vogae FABPs are also phylogenetically related to the FABP3/FABP4 mammalian FABP subfamilies. The native proteins were purified by chromatographical procedures, and apparent molecular mass and isoelectric point were determined. Immunolocalization studies determined the localization of the expression of these proteins in the larval form of the parasite. The genomic exon-intron organization of both genes is also reported, and supports new insights on intron evolution. Consensus motifs involved in splicing were identified.
Assuntos
Evolução Molecular , Proteínas de Ligação a Ácido Graxo/química , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Helminto/química , Proteínas de Helminto/genética , Mesocestoides/metabolismo , Sequência de Aminoácidos , Animais , Éxons , Proteínas de Ligação a Ácido Graxo/isolamento & purificação , Proteínas de Helminto/isolamento & purificação , Íntrons , Microscopia Confocal , Dados de Sequência Molecular , Filogenia , Alinhamento de SequênciaRESUMO
Many parasites undergo sudden changes in environmental conditions at some stage during their life cycle. The molecular response to this variation is characterised by a rapid transcriptional activation of a specific set of genes coding for proteins generically known as stress proteins. They appear to be also involved in various biological processes including cell proliferation and differentiation. The platyhelminth parasite, Mesocestoides corti (Cestoda) presents important properties as a model organism. Under stress conditions, key molecules involved in metabolic pathways as well as in the growth and differentiation of the parasite can be identified. 2D protein expression profile of tetrathyridia of M. corti, submitted to nutritional starvation and cold stress is described, as well as the recovery pattern. A set of specifically expressed proteins was observed in each experimental condition. Quantitative and qualitative differences and stress recovery pattern are also reported. This work makes evident the high plasticity and resistance to extreme environmental conditions of these parasites at the molecular level.
Assuntos
Temperatura Baixa , Proteínas de Choque Térmico/análise , Proteínas de Helminto/análise , Mesocestoides/metabolismo , Animais , Eletroforese em Gel Bidimensional , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Choque Térmico/biossíntese , Proteínas de Choque Térmico/química , Proteínas de Choque Térmico/genética , Proteínas de Helminto/biossíntese , Proteínas de Helminto/química , Proteínas de Helminto/genética , Ponto Isoelétrico , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Masculino , Mesocestoides/genética , Mesocestoides/crescimento & desenvolvimento , Camundongos , Peso Molecular , Proteômica , Coloração pela PrataAssuntos
Echinococcus/crescimento & desenvolvimento , Echinococcus/metabolismo , Proteínas de Helminto/biossíntese , Tropomiosina/biossíntese , Sequência de Aminoácidos , Animais , Antígenos de Helmintos/genética , Antígenos de Helmintos/isolamento & purificação , Echinococcus/genética , Regulação da Expressão Gênica , Proteínas de Helminto/genética , Larva/genética , Larva/crescimento & desenvolvimento , Tropomiosina/genéticaRESUMO
This work describes a new gene coding for a fatty acid binding protein (FABP) in the parasite Echinococcus granulosus, named EgFABP2. The complete gene structure, including the promoter sequence, is reported. The genomic coding domain organisation of the previously reported E. granulosus FABP gene (EgFABP1) has been also determined. The corresponding polypeptide chains share 76% of identical residues and an overall 96% of similarity. The two EgFABPs present the highest amino acid homologies with the mammalian FABP subfamily containing heart-FABPs (H-FABPs). The coding sequences of both genes are interrupted by a single intron located in the position of the third intron reported for vertebrate FABP genes. Both genes are expressed in the protoscolex stage of the parasite. The promoter region of EgFABP2 presents several consensus putative cis-acting elements found in other members of the family, suggesting interesting possible mechanisms involved in the host-parasite adaptation.