Nanoparticle formulation for the development of a dog nanovaccine against Cystic Echinococcosis.

Cystic Echinococcosis is a cosmopolitan zoonosis closely linked to poverty and ignorance. It affects both cattle and humans, causing significant losses to both human and animal health. To date, there is no effective way to combat this. Our proposal focused on the formulation of poly (lactic-co-glycolic acid (PLGA) and Eudragit-RSPO polymeric nanoparticles, which are suitable to encapsulate an antigen for oral administration in dogs. This antigen, named EgFABP1, belonging to the family of fatty acid-binding proteins, was isolated from the larval form of the parasite Echinococcus granulosus. Several reports point to proteins from this family from parasitic flatworms as candidates for a successful vaccine, considering the restricted lipid metabolism of these organisms. The encapsulation of the antigen yielded an efficiency higher than 50 %, and the nanoparticles showed the expected size range. In addition, antigen integrity was conserved and the formulation was resistant to artificial gastric and intestinal fluid effects.

Development of an oral nanovaccine for dogs against Echinococcus granulosus.

Dogs are the main source of animal and human cystic echinococcosis caused by the Cestode parasite Echinococcus granulosus. Dog vaccination seems to be a good strategy to control this parasitic disease. Here we present the development of a polymeric nanoparticle-based oral vaccine for dogs against Echinococcus granulosus delivered in enteric-coated capsules. To achieve our target, we encapsulated two recombinant antigens into biodegradable polymeric nanoparticles in the presence of Monophosphoryl lipid A as an adjuvant to ensure efficient delivery and activation of a protective mucosal immune response. The formulated delivery system showed a nanoparticle size less than 200 nm with more than 80 % antigen encapsulation efficiency and conserved integrity and immunogenicity. The nanoparticle surface was coated with chitosan to enhance adhesion to the gut mucosa and a subsequent antigen delivery. Chitosan-coated nanoparticles showed a higher cell internalization in murine macrophages and dendritic cells as well as a higher penetration into Caco-2 cells in vitro. Antigen-loaded nanoparticles were freeze-dried and enteric-coated capsules were filled with the obtained powder. The obtained results show a promising nanoparticles delivery system for oral vaccination.

Bem-estar vocal de professores: uma proposta de intervenção realizada à distância / Vocal welfare of teachers: a proposal for intervention developed by distance mode learning / Bien-estar vocal de professores: una propuesta de intervención realizada a distancia

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.

Identification of novel CAP superfamily protein members of Echinococcus granulosus protoscoleces.

Echinoccocus granulosus is the causative agent of Cyst Echinococcosis, a zoonotic infection affecting humans and livestock representing a public health and an economic burden for several countries. Despite decades of investigation an effective vaccine still remains to be found. Parasitic cysteine-rich secretory proteins, antigen 5 and pathogenesis-related 1 proteins (CAPs) have been proposed as vaccine candidates against helmith's infection. In this work we have identified two novel proteins of this superfamily expressed at the protoescoleces larval stage named EgVAL1 and EgVAL2. The open reading frame sequences were deduced. The aminoacidic sequence was analyzed and confronted against already known vertebrate' and helminth's proteins sequences in order to infer putative functions. Immunolocalization studies were also performed. The obtained data supported by immunolocalization studies and homology models suggest that these proteins could be involved in protease activity inhibition.

Echinococcus granulosus fatty acid binding proteins subcellular localization.

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.

Voz do professor: fatores predisponentes para o bem-estar vocal / Voz del profesor: los factores predisponentes del bienestar vocal

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...

Reprodutibilidade (teste-reteste) de sintomas vocais e sensações laringofaríngeas / Reproducibility (test-retest) of vocal symptoms and laryngeal-pharyngeal sensations

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...

Direct interaction between EgFABP1, a fatty acid binding protein from Echinococcus granulosus, and phospholipid membranes.

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.

An oral recombinant vaccine in dogs against Echinococcus granulosus, the causative agent of human hydatid disease: a pilot study.

Dogs are the main source of human cystic echinococcosis. An oral vaccine would be an important contribution to control programs in endemic countries. We conducted two parallel experimental trials in Morocco and Tunisia of a new oral vaccine candidate against Echinococcus granulosus in 28 dogs. The vaccine was prepared using two recombinant proteins from adult worms, a tropomyosin (EgTrp) and a fibrillar protein similar to paramyosin (EgA31), cloned and expressed in a live attenuated strain of Salmonella enterica serovar typhimurium.In each country, five dogs were vaccinated with the associated EgA31 and EgTrp; three dogs received only the vector Salmonella; and six dogs were used as different controls. The vaccinated dogs received two oral doses of the vaccine 21 d apart, and were challenged 20 d later with 75,000 living protoscoleces. The controls were challenged under the same conditions. All dogs were sacrificed 26-29 d postchallenge, before the appearance of eggs, for safety reasons.We studied the histological responses to both the vaccine and control at the level of the duodenum, the natural localization of the cestode. Here we show a significant decrease of parasite burden in vaccinated dogs (70% to 80%) and a slower development rate in all remaining worms. The Salmonella vaccine EgA31-EgTrp demonstrated a high efficacy against E. granulosus promoting its potential role in reducing transmission to humans and animals.

Two novel Mesocestoides vogae fatty acid binding proteins--functional and evolutionary implications.

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.

Genomic structure and expression of a gene coding for a new fatty acid binding protein from Echinococcus granulosus.

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.