Riscos laborais associados à manipulação de nanomateriais

De um modo geral, os nanomateriais manufaturados (NM) são definidos como materiais fabricados deliberadamente e que contêm partículas com pelo menos uma dimensão externa na gama de tamanhos compreendida entre 1 e 100 nanómetros (Comissão Europeia, 2011). A sua pequena dimensão confere-lhes propriedades físicas, químicas e biológicas que podem diferir bastante das propriedades dos materiais com a mesma composição química mas utilizados numa escala não nanométrica. São as propriedades mecânicas, óticas, elétricas e magnéticas inerentes aos materiais na escala “nano” que os tornam vantajosos para as mais diversas aplicações industriais e biomédicas. Contudo, a enorme expansão que tem vindo a acontecer ao nível da síntese, produção industrial e utilização de NMs contrasta com uma ainda insuficiente avaliação de risco para a saúde humana e para o ambiente. Efectivamente, A European Agency for Safety and Health at Work (EU-OSHA,2009) considerou a exposição a NM como o risco emergente mais premente no contexto da saúde ocupacional, estimando que entre 300.000 a 400.000 postos de trabalho lidavam já directamente com as nanotecnologias. Neste seminário será abordada a possibilidade de exposição ocupacional ao longo do ciclo de vida dos NM, bem como as suas potenciais implicações para a saúde dos trabalhadores. Nesta palestra são apresentados alguns estudos realizados no Instituto Nacional de Saúde Doutor Ricardo Jorge que produziram evidência científica que poderá contribuir para o esforço internacional da regulação da produção e aplicação de nanomateriais, salvaguardando a saúde humana face às suas aplicações inovadoras.

Conservação de material genético de espécies silvestres do bioma caatinga utilizando a manipulação oócitos inclusos em folículos ovarianos pré antrais (MOIFOPA)

The objective of the present thesis was to use the manipulation of oocytes enclosed in preantral follicles (MOEPF) as a tool for the female gametes rescue and optimization, from wild species of Caatinga biome. The thesis was divided into 4 experiments. At first experiment, it was performed the estimative and description of the agouti (Dasyprocta leporina) preantral follicles (PF) histologic and ultrastructural features, in which it was estimated 4419.8 ± 532.26 and 5397.52 ± 574.91 follicles for the right and left ovary, respectively, and the majority (86,63%) belonged to the primordial follicles category (P<0.05). Most of the population consists of morphologically normal follicles (70.78%), presenting a large and central nuclei and uniform cytoplasm. At ultrastructural evaluation it was verified the presence of a great number of round mitochondrias associated to lipid droplets. In the second experiment, it was performed the estimative and description of yellow-toothed cavies (Galea spixii) PF characteristics, also, the evaluation of the effect of solid surface vitrification (SSV) on the in situ PF morphology. The total of 416.0 ± 342.8 PF was estimated for the ovary pair and the presence of a large quantity of primary follicles (P<0.05) was evidenced. Most of the PF was morphologically normal (94.6%), in which the oocyte nuclei presented condensed granules of heterochromatin. Round or elongated shaped mitochondria constituted the most abundant organelles. In regard of the SSV, the protocol using the dimethylsulfoxide (DMSO) 3M possibility the preservation of 69.5% of morphologically normal PF, which was evidenced by the light and transmission electronic microscopy. At third experiment, the evaluation of the SSV procedure on the morphology and viability in situ PF form collared peccaries (Pecari tajacu) was performed. No differences were observed among treatments, in which the use of DMSO, ethylene glycol (EG) and dimethylformamide (DMF) as cryoprotectants, regardless its concentration, promoted the morphology preservation of much than 70% of PF. Concerning the PF viability, the DMSO and EG promoted the best preservation. The fourth experiment aimed to evaluate the effect of α MEM+ or TCM199 associated or not to 50 ng of FSHr on the morphology, activation and growth of collared peccaries PF, in vitro cultured (IVC) during 1 or 7 days and the effect on the extracellular matrix (ECM). After 7 days of IVC only the use of TCM199/FSH maintained the proportion of intact PF, similar to day 1(63.2%), however, no differences were observed among treatments (P>0.05). Also, an improvement of the proportion of intact growing PF was verified (P>0.05). By the Ag-NOR analysis it was observed that only the treatment using TCM199/FSH promoted the maintenance of cell proliferation similar to day 1 (P>0.05). The picrosirius red stain revealed that ECM remained intact in all treatments (P>0.05). Thus, as the general conclusion, the use of MOEPF in the refereed species allowed the knowledge of aspects related to its reproductive morphology and physiology, enabling the germplasm conservation, with the possibility of germplasm bank formation, as the elucidation of mechanisms related to the PF survive and in vitro development.

Plasticidade dependente da experiência induzida por manipulação da matriz extracelular

The primary somatosensory cortex (S1) receives inputs from peripheral tactile receptors and plays a crucial role on many important behaviors. However, the plastic potential of this region is greatly reduced during adulthood, limiting functional recovery after injuries. This fact is due to the presence, in the brain parenchima, of structures and substances that have an inhibitory effect on plasticity, such as chondroitin sulfate proteoglicans (CSP) present in the perineuronal.nets (PNNs) surrounding a subset of neurons. Maturation of PNNs coincide with the closure of critical periods of plasticity in cortical areas, since CSP act to stabilize synaptic contacts. Removal of CSP is proven to be an effective therapeutic approach to restore plasticity and increase the odds of functional recovery after cortical lesion. In the present work, we removed CSP from the sensorimotor cortex of rats to restore plasticity and promote the compensatory morphofunctional regeneration of cortical circuits modified by removal of mystacial vibrissae during the critical period. Treatment with the CSP-digesting enzyme chondroitinase ABC proved efficient to restore plasticity in S1 circuits, as evidenced by morphological rearrangements and functional recovery.

Plasticidade dependente da experiência induzida por manipulação da matriz extracelular

The primary somatosensory cortex (S1) receives inputs from peripheral tactile receptors and plays a crucial role on many important behaviors. However, the plastic potential of this region is greatly reduced during adulthood, limiting functional recovery after injuries. This fact is due to the presence, in the brain parenchima, of structures and substances that have an inhibitory effect on plasticity, such as chondroitin sulfate proteoglicans (CSP) present in the perineuronal.nets (PNNs) surrounding a subset of neurons. Maturation of PNNs coincide with the closure of critical periods of plasticity in cortical areas, since CSP act to stabilize synaptic contacts. Removal of CSP is proven to be an effective therapeutic approach to restore plasticity and increase the odds of functional recovery after cortical lesion. In the present work, we removed CSP from the sensorimotor cortex of rats to restore plasticity and promote the compensatory morphofunctional regeneration of cortical circuits modified by removal of mystacial vibrissae during the critical period. Treatment with the CSP-digesting enzyme chondroitinase ABC proved efficient to restore plasticity in S1 circuits, as evidenced by morphological rearrangements and functional recovery.

Manipulação gênica do protozoário Neospora caninum como ferramenta para o estudo das interações parasito-hospedeiro

Neospora caninum is an obligate intracellular parasite classified in the phylum Apicomplexa, characterized by the presence of the apical complex composed by micronemes proteins, rhoptries and dense granules, used by parasite during the adhesion and invasion process of the host cell. This is the mean event in infection pathogenesis generated by N. caninum and other parasites from the phylum Apicomplexa, promoting influence in the parasite biology and the interface between the parasite and its host. Therefore, molecular tools have been developed in order to identify and characterize these possible virulence factors. Thus, the present study sought to establish a specific system of genetic manipulation of N. caninum, searching for the improvement of the genetics manipulation of this parasite. So, we developed genetically depleted N. caninum to Rop9 rhoptry using the pU6-Universal CRISPR-Cas9 plasmid of T. gondii modified by the insertion of Ku80. The Rop9 depleted parasite showed important during initial phase of invasion and replication of the parasite, however it was not characterized as a potential virulence fator for N. caninum. Furthermore, T. gondii proteins were expressed in N. caninum by the use of specific vectors for this parasite, showing an heterologous system for the study of Toxoplasma proteins, due to the fact that Gra15 or Gra24 of type II T. gondii and Rop16 of type I T. gondii were expressed in N. caninum tachyzoites in a stable way and keept its biological phenotype, as already presented the former parasite, that naturaly expresses these proteins. In addition, it was observed that N. caninum induced an inflammasome activation through NLRP3, ASC and Caspase-1. IL-1R/MyD88 demonstrated an indirect pathway in the control of parasite replication. Furthermore, it was observed that this activation is dependent of the potassium efflux and that different strains of N. caninum keep this activation profile. However, T. gondii strains block this activation, making necessary a prior signal in order to active the inflamosome pathway. Type I T. gondii Rop16 was identified as responsible for blocking this activation, in a dependent way to the STAT3 activation. Therefore, the development of molecular tools and their application in N. caninum may prove to be useful to identify and characterize virulent factors involved in the pathogenesis by these two protozoans.

Luz-personagem: o jogo de manipulação da iluminação cênica em cenas cômicas

This research has as a central point of discussion the relationship between the action and the stage lighting, directed to scenes in which, from the exchange between actor and professional lighting, you can develop comic situations. The particularity of these scenes is the involvement of lighting as presentification feature a character in the scene, made possible by game created with the actor. In order to develop this discussion, the author divides the text into three chapters, the first being a memorial of his artistic career and his creative processes, enabling the reader to situate the paths taken by the author for the study. Then discusses possible relationship between performance and lighting, pointing situations where the two are directly related. Finally, an analysis and comparison of two comic scenes in which the actor play with the lighting suggests a character through presentification lighting on the scene. Leads the discussion to the conclusion, proposing the term light-character to set the lighting feature, suggesting characters in the scene.

Contributo do sistema integrado de comunicação e manipulação no desenvolvimento e aprendizagem de crianças com paralisia cerebral em contexto educativo/escolar

Dissertação apresentada à Escola Superior de Educação de Lisboa para obtenção do grau de mestre em Educação Especial – ramo de Problemas de Cognição e Multideficiência

O ator que canta um conto: a manipulação de parâmetros musicais na voz do ator

This research started with an investigation about the theatrical speaking. Through an empirical methodology that analyzed a practical experience of creating a play and bibliographical research, the actress/researcher investigated ways to manipulate musical parameters as tools in the creation process of the actor s voice. The actress/researcher attempted to connect theory and practice, moved by the desire to find a vocal expression in theater that unfolds as living and transforming movement. This dissertation also contains the report of pedagogical experiences, in which the actress/researcher explored strategies to teach the appropriation of musical parameters in the construction of the vocal work of the actor. Considering that speaking in theater is closer to singing than everyday speech, she concluded that the actor may compose music as a music composer does in the elaborating process of building vocal scores. Therefore, she demonstrated that it is of fundamental importance a musical training in the development of the actor.

O ator que canta um conto: a manipulação de parâmetros musicais na voz do ator

This research started with an investigation about the theatrical speaking. Through an empirical methodology that analyzed a practical experience of creating a play and bibliographical research, the actress/researcher investigated ways to manipulate musical parameters as tools in the creation process of the actor s voice. The actress/researcher attempted to connect theory and practice, moved by the desire to find a vocal expression in theater that unfolds as living and transforming movement. This dissertation also contains the report of pedagogical experiences, in which the actress/researcher explored strategies to teach the appropriation of musical parameters in the construction of the vocal work of the actor. Considering that speaking in theater is closer to singing than everyday speech, she concluded that the actor may compose music as a music composer does in the elaborating process of building vocal scores. Therefore, she demonstrated that it is of fundamental importance a musical training in the development of the actor.

Association of insulin resistance and GLP-2 secretion in obesity: a pilot study

OBJECTIVE: The objective of this pilot study was to determine whether glugagon-like peptide 2 (GLP-2) secretion relates to insulin sensitivity (IS) in obese subjects. SUBJECTS AND METHODS: Twenty four obese subjects [body mass index (BMI) 40.0 ± 3.0 kg/m² (mean ± standard deviation)] were included, nine of which were male, age 43 ± 8 years. Twelve subjects had type 2 diabetes, all treated with oral anti-diabetic agents only. The subjects were submitted to standard meal tolerance test (MTT) for dosage of the curves: glucose, insulin, and GLP-2. Insulin sensitivity was measured by HOMA-IR, and OGIS was derived from the MTT. Spearman linear correlations and partial correlations were obtained. RESULTS: There was an inverse relationship between the GLP-2 secretion and IS: HOMA-IR correlated with GLP-2 AUC (R = 0.504; p = 0.012), and OGIS correlated with GLP-2 incremental AUC (R = -0.54; p = 0.054). The correlation persisted after controlling for BMI. CONCLUSION: We found an association of GLP-2 secretion and insulin resistance (IR). The understanding of the underlying mechanisms may provide future directions in the pharmacological manipulation of incretins, and in the treatment of obesity and related metabolic disorders.

Cinética de remoção de lactato na definição de pausas para treinamento intervalado de alta intensidade

Universidade Estadual de Campinas . Faculdade de Educação Física

Dopamina, óxido nítrico e suas interações em modelos para o estudo da esquizofrenia

Modelos experimentais baseados no aumento da neurotransmissão dopaminérgica mimetizam aspectos comportamentais e neuroquímicos característicos da esquizofrenia. Psicoestimulantes, como a anfetamina, são utilizados com esta finalidade, pois aumentam os níveis de dopamina extracelular nas vias mesocorticolímbica e mesoestriatal. As limitações da manipulação direta do sistema dopaminérgico nos modelos animais incentivam abordagens complementares. O óxido nítrico (NO), um neurotransmissor atípico que inibe a recaptação de dopamina e estimula sua liberação, parece modular comportamentos controlados pelo sistema dopaminérgico. O teste de inibição pré-pulso revela uma deficiência no filtro sensório-motor, verificada em esquizofrênicos ou após tratamentos com psicotomiméticos, podendo ser prevenida pela inibição do NO. Esta revisão apresenta evidências da interação do NO com o sistema dopaminérgico em modelos para o estudo da esquizofrenia como uma nova ferramenta de investigação desta patologia.