A marcha humana: circuitos neuronais entre estruturas corticais vs sub-corticais

As conexões entre os níveis corticais e sub-corticais na activação da marcha Humana carecem de discussão. As alterações da marcha por lesão no território da Artéria Cerebral Média (ACM) podem ser explicadas pela disfunção de circuitos neuronais dos Núcleos da Base ao córtex e Núcleos Pedúnculo-Pontino. Este trabalho tem como objectivo identificar os percursos anatómicos das principais conexões entre as estruturas encefálicas referidas no território da ACM. Com base na topografia das conexões neuronais, é aceitável que as alterações da marcha sejam explicadas por alterações na função dos Núcleos da Base, através das suas conexões, enquanto moduladores da actividade motora.The connections between cortical and sub-cortical levels in human gait activation require discussion. The gait alterations after a lesion in the territory of the Middle Cerebral Artery (MCA) can be explained by the dysfunction of neuronal circuits of the basal ganglia to the cortex and peduncle-pontine nuclei. The purpose of this work is to identify the anatomical pathways of the main connections between the brain structures mentioned on MCA territory. Based on the topography of neuronal connections it´s acceptable that gait alterations may also be due to dysfunctions in basal ganglia through their connections, as modulators of motor activity

Strategies of plasmid DNA production and their influence on therapeutic applications

Current developments in gene therapy and DNA vaccination, plasmid DNA vectors are becoming increasingly appealing as therapeutics towards a large number of diseases such as cancer, infectious and cardiovascular diseases. This popularity is creating the demand for high quantities of highly purified plasmid DNA which in turn requires the design of high pDNA yield bioprocesses. However, opposed to recombinant protein production, research on pDNA production is still needed, in order to have a clear comprehension of all the challenges and bottlenecks faced during the production of plasmid DNA (pDNA). The design of a plasmid DNA production process usually begins with the choice of a suitable culture medium to cultivate the expression system containing the therapeutic plasmid. After defining all medium components, the influence of culture conditions, such as pH, temperature and dissolved oxygen, on biomass and plasmid yields is generally studied. Since the appropriate conditions for maximizing biomass production and plasmid replication are not usually the same, a compromise solution is usually considered in these cases. When designing a large scale plasmid DNA production process, the employment of a correct fermentation strategy is also necessary in order to improve yields while reducing production costs. So far, reports on plasmid DNA production in E. coli are focused essentially on the influence of different medium composition, fermentation conditions and feeding strategies on overall biomass and pDNA mass and volumetric titres. Nevertheless, due to the complex nature of microbial growth and the application of several modes of operation such as batch, fed-batch and continuous cultivations, the constant monitoring and control of pDNA bioprocesses represents an engineering challenge that should not be disregarded. As a highest yield process may not correspond necessarily to the best fermentation design, the improvement in off-line, at-line and online monitoring techniques should be seen as a crucial task in the design of the fermentation. Two of the most relevant factors for fermentation performance are the existence of host cell metabolic stress and plasmid instability; hence, the characterization of cell physiology and plasmid segregational stability has to be considered and monitored during the process. With this thesis, we attempted to improve plasmid DNA yields while gaining new insights on plasmid DNA fermentation processes through the use of novel monitoring techniques such as flow cytometry and real-time quantitative PCR. We started this work studying the influence of growth temperature and tryptone concentration on plasmid DNA production in a previously developed semi-defined medium. The analysis of pDNA yields and E. coli morphology revealed that higher pDNA specific yields were obtained at higher temperatures (37 and 40 ºC). Also, at these temperatures, E. coli filamentation was observed, possibly indicating a higher metabolic stress due to higher plasmid replication and higher culture temperatures. When analyzing the influence of tryptone concentration on plasmid yield, the best results were achieved with the lowest tryptone concentration used. The use of limiting tryptone concentrations at a temperature of 37 ºC was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure (supercoiled isoform); thus favouring the attainment of product quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved, proving that this strategy is able to increase pDNA yield even at small scale. Since this first study revealed some evidence of E. coli metabolic stress during cultivation, the next task consisted in the development of new flow cytometric methodologies that allowed single cell physiology monitoring during cultivation for a better understanding of cultivation conditions influence in the host metabolic activity. Because one of the parameters that enable a better characterization of cell metabolic cell as well as population heterogeneity is cell cycle progression, in the second part of the work, we developed a flow cytometric method to evaluate cell cycle progression in E. coli cultivation using a newly developed far-red dye, DRAQ5. In this study we demonstrated that the use of DRAQ5 as a DNA-specific labelling stain provided an easy assessment of intracellular DNA content and cell-cycle phases in the Gram-negative bacteria E. coli. Besides the previously reported method for cell cycle analysis, another method for assessing cell viability was implemented using flow cytometry. In this method, a propidium iodide/bis- (1,3-dibutylbarbituric acid)trimethine oxonol (BOX) dual staining was used to distinguish between three different populations: healthy cells (no staining), cells with depolarized membranes (stained with BOX) and cells with permeabilized membranes (stained with propidium iodide and BOX). In order to evaluate plasmid copy number (PCN) throughout the fermentation, a real-time quantitative PCR method was developed for absolute plasmid copy number quantification in whole E. coli cells. After developing and implementing these methods, we evaluated the impact of several plasmid DNA induction strategies, such as amino acid limitation, AMP addition and temperature up-shift, on cell physiology and plasmid segregational stability. This study showed that all induction strategies caused cell filamentation, due to an increase in forward scatter values, and decreased viability at the end of fermentation, as was seen by an increase in the percentage of depolarized and permeabilized cells. The results also suggest that an amino acid limitation with AMP addition induction strategy resulted in the highest specific yields and, concomitantly, highest PCN values. In conclusion, amino acid limitation-based amplification strategies seemed to be suitable approaches to be implemented at a large scale level since they do not require any additional energy and also had proved to be efficient in plasmid amplification, without causing any detrimental effects in plasmid stability and cellular viability. The last step of this work aimed at improving plasmid DNA yield through the study of different batch and fed-batch fermentations in bioreactors. Also, the influence of different glycerol and tryptone concentrations and different non-feedback feeding profiles, namely exponential and constant feed rates, on cell physiology and plasmid stability was evaluated by means of flow cytometry and real-time qPCR, respectively; investigating the potential of these two techniques as valuable tools for bioprocess monitoring and design. The results showed that all fermentation strategies caused a slight decrease of cell viability at the end of fermentation, being this decrease more pronounced in fed-batch fermentations than in batch fermentations. The time-course assessment of plasmid copy number revealed that PCN values suffered an increase at the end of batch fermentations, which is in agreement with our previous results obtained in batch fermentations performed in shake flasks. However, in fedbatch fermentations, there were pronounced fluctuations in PCN values throughout the fermentations, indicating some plasmid segregational instability. As supposed, fed-batch fermentations with exponential or constant feeding profiles yielded higher biomass and plasmid DNA than batch fermentations with the highest biomass and plasmid yields being obtained with a fed-batch strategy with an exponential feed rate of 0.2 h-1. Notwithstanding the high biomass (95.64 OD600) and plasmid yields (344.30 mg pDNA/L) obtained, this fermentation also exhibited higher plasmid instability and lower percentage of viable cells. This work showed that the fermentation strategy used, not only influences product yield, but also cell physiology and pDNA segregational stability. Furthermore, the new findings described herein draw attention towards the relevance of monitoring bioprocess performance and not just overall biomass and product yields. In conclusion, in this thesis we evaluated and improved pVAX1-LacZ plasmid production in Escherichia coli DH5 alpha taking into account not only the overall biomass and plasmid yields, but also considering that cell physiology and plasmid segregational stability, that are two pivotal features to the design and development of these production bioprocesses. In order to study these two factors, several techniques were implemented and were later used to evaluate the influence of several fermentation parameters such as induction strategies and fermentation strategies on overall process performance.Work financed by the Portuguese Foundation for Science and Technology (SFRH/BD/41521/2007) under the programe QREN - POPH - Type 4.1 –Advanced Training, cofunded by the European Social Fund and by national funds from the MCTES

Severidade e prognóstico no acidente vascular encefálico: revisão scoping [artigo]

O acidente vascular encefálico (AVE) tem vindo a apresentar uma taxa de incidência estável e um considerável declínio na taxa de mortalidade, o que corresponde a um aumento na prevalência de sobreviventes. O conhecimento das alterações funcionais que podem surgir após o AVE, da sua severidade e das estratégias disponíveis para avaliar a disfunção, facilita a construção de um plano de reabilitação, com objetivos para os profissionais de saúde, para os pacientes e para a família dentro do potencial de recuperação. A severidade surge, como um conceito abrangente associado à presença de deficiências das estruturas, deficiências das funções, limitações das atividades e restrições da participação social. Deficiências, limitações e restrições mais severas fazem prever uma recuperação mais difícil e mais prolongada. A determinação do prognóstico em indivíduos com AVE engloba não só o risco de morte a curto prazo, como também a probabilidade de recuperar a função a longo prazo. Avaliar o panorama acerca da informação existente sobre o nível de severidade e prognóstico em AVE. A revisão scoping baseou-se na metodologia de Arksey & O’Malley (2005), sendo constituída por seis passos: 1) identificação da questão; 2) identificação da literatura relevante; 3) seleção da literatura; 4) mapeamento dos dados; 5) recolha, sumário e transcrição dos resultados; 6) consultoria (opcional). Foram analisados 47 estudos observacionais. Noventa e cinco por cento dos autores referem-se à severidade como sendo a quantidade de défices neurológicos apresentados pelos indivíduos após o AVE e avaliam-na através de instrumentos de medida específicos para a avaliação de défices neurológicos (76% dos autores utilizaram a NIHSS na sua metodologia). O prognóstico no AVE surge associado à funcionalidade afetada (89%); probabilidade/índice de mortalidade (54%); e encaminhamento após a alta (15%). O prognóstico pode ser influenciado por fatores pessoais e ambientais, fatores clínicos e por algumas comorbilidades, entre outros. Os estudos de severidade e prognóstico em AVE poderão não refletir a condição real do indivíduo e induzir em erro a aplicação destes conceitos na prática clínica, influenciando o prognóstico esperado.info:eu-repo/semantics/publishedVersio

Radiology Case

Lactente do sexo masculino com seis meses. Antecedentes pessoais: gestação de termo, vigiada e sem intercorrências, somatometria ao nascimento adequada, índice de Apgar 10/10 no 1º e 5º minuto e período neonatal sem intercorrências.

Ajustes posturais antecipatórios no movimento de alcançar em indivíduos pós-acidente vascular encefálico

Pretendeu-se verificar a existência de alterações nos timings de activação dos músculos estabilizadores da omoplata em indivíduos pós-acidente vascular encefálico e a sua relação com a velocidade. Metodologia: Amostra (n = 17) dividida nos Grupos 1 (n = 10 indivíduos sem patologia) e 2 (n = 7 indivíduos pós-acidente vascular encefálico). Análise electromiográfica e cinemática no alcance de um copo. Resultados: Nas fibras inferiores do trapézio esquerdo (p = 0,043) e grande dentado bilateralmente (p = 0,028), encontraram-se diferenças entre os Grupos. Não se verificou uma relação com a velocidade. Conclusões: Foram encontrados ajustes posturais antecipatórios em alguns dos músculos.This study aimed to verify the existence of impairments at the activation timings of the scapular stabilizers in post-stroke subjects and their relation to the velocity. Methods: The sample (n = 17) was divided in Groups 1 (n = 10 healthy subjects) and 2 (n = 7 post-stroke subjects). Electromyographic and kinematic analysis was performed during reaching. Results: Differences between the groups were found at the inferior fibers of the left trapezius (p = 0,043) and serratus anterior bilaterally (p = 0,028). No relation was found to the velocity. Conclusions: In some muscles were found anticipatory postural adjustments

Reliability of two methods for identifying the postural phase of gait initiation in healthy and post-stroke subjects

This study aims to compare two methods of assessing the postural phase of gait initiation as to intrasession reliability, in healthy and post-stroke subjects. As a secondary aim, this study aims to analyse anticipatory postural adjustments during gait initiation based on the centre of pressure (CoP) displacements in post-stroke participants. The CoP signal was acquired during gait initiation in fifteen post-stroke subjects and twenty-three healthy controls. Postural phase was identified through a baseline-based method and a maximal displacement based method. In both healthy and post-stroke participants higher intra-class correlation coefficient and lower coefficient of variation values were obtained with the baseline-based method when compared to the maximal displacement based method. Post-stroke participants presented decreased CoP displacement backward and toward the first swing limb compared to controls when the baseline-based method was used. With the maximal displacement based method, there were differences between groups only regarding backward CoP displacement. Postural phase duration in medial-lateral direction was also increased in post-stroke participants when using the maximal displacement based method. The findings obtained indicate that the baseline-based method is more reliable detecting the onset of gait initiation in both groups, while the maximal displacement based method presents greater sensitivity for post-stroke participants

Em tempos de visitas : inquisição, circulação e oralidade escrava na Bahia (1590-1620)

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Humanas, Programa de Pós-Graduação em História, 2014.Com base nos registros de visitações do Santo Ofício à capitania da Bahia, este estudo formula uma análise sobre o lugar dos escravos na circulação de informações na Bahia do final do século XVI e início do século XVII. Expõe o modo como estavam normatizadas as visitações por meio dos regimentos de 1552 e 1613 e qual o uso feito deste corpo de normas no cotidiano das pessoas que habitavam esse espaço. Aponta para a existência de uma agência escrava sustentada por meio de uma circulação de pessoas e informações que geravam boatos e rumores até chegar à mesa da Inquisição. Investiga as interações entre os membros dessa sociedade pautando as relações entre os vizinhos, escravos, cristãos-velhos e cristãos-novos. Nesse sentido, tanto como acusadores, quanto acusados, os escravos em sua agência tornaram-se importante vetor de comunicação utilizado pela comunidade baiana e Inquisição, o que demonstra certa complexidade em relação à circulação de informações e ideias existentes na época. _______________________________________________________________________________________ ABSTRACTBased on the records of visitations of the Holy Office to the captaincy of Bahia, this study makes an analysis of the hole of slaves in the stream of information, from late sixteenth and early seventeenth century. It outlines how the visitations were normalized by the regiments of 1552 and 1613 and how this body of standards was engaged in the daily lives of local people. This text also points the existence of a slave agency suported by a movement of people and information, a grid of gossip and rumors that used to end on Inquisition's table. The interactions between the members of that society, mainly relations between the neighbors, slaves, Old Christians and New Christians, are also investigate for the proposes of this text. In this sense, as accusers and as accused as well, the slaves by their agency became important vector of local communication used by the Inquisition and Bahian community, this shows the complexity of the stream of information and ideas that existed in that time

Interlimb relation during the double support phase of gait: an electromyographic, mechanical and energy based analysis

The purpose of this study is to analyse the interlimb relation and the influence of mechanical energy on metabolic energy expenditure during gait. In total, 22 subjects were monitored as to electromyographic activity, ground reaction forces and VO2 consumption (metabolic power) during gait. The results demonstrate a moderate negative correlation between the activity of tibialis anterior, biceps femoris and vastus medialis of the trailing limb during the transition between midstance and double support and that of the leading limb during double support for the same muscles, and between these and gastrocnemius medialis and soleus of the trailing limb during double support. Trailing limb soleus during the transition between mid-stance and double support was positively correlated to leading limb tibialis anterior, vastus medialis and biceps femoris during double support. Also, the trailing limb centre of mass mechanical work was strongly influenced by the leading limbs, although only the mechanical power related to forward progression of both limbs was correlated to metabolic power. These findings demonstrate a consistent interlimb relation in terms of electromyographic activity and centre of mass mechanical work, being the relations occurred in the plane of forward progression the more important to gait energy expenditure

Ankle anticipatory postural adjustments during gait initiation in healthy and post-stroke subjects

Anticipatory postural adjustments during gait initiation have an important role in postural stability but also in gait performance. However, these first phase mechanisms of gait initiation have received little attention, particularly in subcortical post-stroke subjects, where bilateral postural control pathways can be impaired. This study aims to evaluate ankle anticipatory postural adjustments during gait initiation in chronic post-stroke subjects with lesion in the territory of middle cerebral artery. Methods: Eleven subjects with post-stroke hemiparesis with the ability to walk independently and twelve healthy controls participated in this study. Bilateral electromyographic activity of tibialis anterior, soleus and medial gastrocnemius was collected during gait initiation to assess the muscle onset timing, period of activation/deactivation and magnitude of muscle activity during postural phase of gait initiation. This phase was identified through centre of pressure signal. Post-stroke group presented only half of the tibialis anterior relative magnitude observed in healthy subjects in contralesional limb (t=2.38, p=0.027) and decreased soleus deactivation period (contralesional limb, t=2.25, p=0.04; ipsilesional limb, t=3.67, p=0.003) as well its onset timing (contralesional limb, t=3.2. p=0.005; ipsilesional limb, t=2.88, p=0.033) in both limbs. A decreased centre of pressure displacement backward (t=3.45, p=0.002) and toward the first swing limb (t=3.29, p=0.004) was observed in post-stroke subjects. These findings indicate that chronic post-stroke subjects with lesion at middle cerebral artery territory present dysfunction in ankle anticipatory postural adjustments in both limbs during gait initiation

Ações anticorrupção no Brasil : previsão normativa e efeitos causados pela Operação Lava-jato no campo político, econômico e social em todo país

Trabalho de Conclusão de Curso (especialização)—Universidade de Brasília, Faculdade de Economia, Administração, Contabilidade e Gestão de Políticas Públicas, Departamento de Administração, Curso de Especialização em Gestão Pública Municipal, 2019.Contexto: O Brasil nos últimos anos vem sendo tomado por escândalos que envolvem a temática corrupção, onde os campos político e econômico do país tornam-se fragilizados diante da quantidade de atos ilícitos cometidos. Objetivo: analisar as normas brasileiras existentes relacionadas ao combate à corrupção e a decisão do agente diante de um ato corrupto. Metodologia: Trata-se de uma pesquisa bibliográfica e descritiva, que terá como fonte livros, jurisprudências, doutrinas, artigos e revistas. Considerações finais: Vale ressaltar que é possível evidenciar que na legislação brasileira existem leis que visam coibir os atos de corrupção. Estes instrumentos buscam, de todos os modos, cercarem e proverem uma administração pública que respeite a dignidade da pessoa humana, por meio de políticas públicas eficazes, além de mitigar práticas de corrupção