Dissertação (mestrado)—Universidade de Brasília, Faculdade de Economia, Administração e Contabilidade, Programa de Pós-Graduação em Administração, Mestrado Profissional em Administração, 2016. / Submitted by Camila Duarte (camiladias@bce.unb.br) on 2017-01-23T13:43:48Z No. of bitstreams: 1 2016_RodrigoFáveroClemente.pdf: 2075366 bytes, checksum: 0e0e111e007e78805c4718605f30d768 (MD5) / Approved for entry into archive by Ruthléa Nascimento(ruthleanascimento@bce.unb.br) on 2017-02-20T17:04:01Z (GMT) No. of bitstreams: 1 2016_RodrigoFáveroClemente.pdf: 2075366 bytes, checksum: 0e0e111e007e78805c4718605f30d768 (MD5) / Made available in DSpace on 2017-02-20T17:04:01Z (GMT). No. of bitstreams: 1 2016_RodrigoFáveroClemente.pdf: 2075366 bytes, checksum: 0e0e111e007e78805c4718605f30d768 (MD5) / A responsividade democrática pressupõe que, em regimes democráticos, governos constituídos devem buscar correspondência entre suas ações e as expectativas e preferências dos governados. Além das formas tradicionais de participação política, foram estabelecidos recentemente no Brasil espaços institucionais mediante os quais a população pode informar, diretamente ou por meio de representantes, expectativas ou preferências aos governantes. A efetividade desses espaços, chamados de instituições participativas, depende da correspondência entre preferências da sociedade e atos do governo. Nesse contexto, a presente dissertação teve por objetivo identificar a efetividade das Conferências Nacionais de Saúde (CNS), realizadas entre os anos de 1996 e 2011, mediante a análise da responsividade do governo federal diante das propostas aprovadas nessas CNS. Para esse fim, estabeleceu-se como premissa que as propostas constantes nas CNS representam a opinião da sociedade em relação à saúde, devendo ser consideradas pelo governo federal no estabelecimento das políticas públicas voltadas à saúde. Para realizar a pesquisa empírica, as propostas foram categorizadas por temas, mediante análise de conteúdo, e posteriormente pareadas com os atos administrativos do governo federal voltados à saúde. Dessa forma foi possível a realização de testes de congruência, método pelo qual a responsividade democrática pode ser mensurada. Com base no referencial teórico utilizado, foi testado empiricamente um conjunto de hipóteses a respeito da responsividade do governo federal e a efetividade das CNS. A análise do conteúdo das propostas revelou que as CNS variaram pouco em relação aos temas tratados. Em relação aos testes de congruência, percebeu-se que a efetividade das CNS variou de acordo com a atuação dos governos. Essa variação apresentou-se não apenas em relação ao número de propostas respondidas, mas sobretudo em relação aos temas que apresentaram congruência. Na parte final da presente dissertação são propostas possíveis explicações para os resultados encontrados, bem como uma agenda de estudos futuros a respeito do tema. / The democratic responsiveness assumes that, in democratic regimes, constituted governments should seek correspondence between their actions and the expectations and preferences of the governed. In addition to traditional forms of political participation, institutional spaces through which the public can inform, directly or through representatives, expectations or preferences to the rulers have been established recently in Brazil. The effectiveness of these spaces, called Participatory institutions, depends on the correspondence between preferences of society and governmental actions. In this context, the present work aimed to identify the effectiveness of the National Conferences of Health (CNS), made between the years 1996 and 2011, through the analysis of federal government’s responsiveness on the proposals approved by these CNS. To this end, it established the premise that the subjects contained in the CNS’s proposals represent the views of society in relation to health, and as so should be considered by the federal government in the establishment of public policies on health. To conduct this empirical research, proposals were categorized by themes, through content analysis, and then paired with the administrative acts of the federal government focused on health. Thus it was possible to perform matching tests, method by which democratic responsiveness can be measured. Based on the theoretical framework, was empirically tested a set of assumptions about the responsiveness of the federal government and the effectiveness of the CNS. The content analysis of the proposals revealed that CNS varied little in relation to the topics covered. Regarding the congruence tests, it was noted that the effectiveness of CNS varied according to the actions of governments. This variation presented not only in the number of answered proposals, but especially for those issues that presented congruence. In the final part of present work some possible explanations are proposed for the findings, as well as an agenda for future studies on the subject.

In my diploma thesis I deal with the issues of hyperactivity that has recently been a very current and discussed topic. In the theoretic part of my thesis, I specify the concept of hyperactivity and talk further about its symptoms and consequent effects. I, as well, pay close attention to the current possibilities of reeducation with a consequent focus on the programs supporting hyperactive children and their parents. The practical part of the diploma thesis consists of three parts: an interview with a pedagogue working with hyperactive children; a case report of a hyperactive child; questionnaire survey and its subsequent interpretation. The practical part of the diploma thesis focuses on the particular symptoms of hyperactivity at hyperactive children in the school environment and their consequent effects. It tries to find out the possibilities of the reeducation and remedy at the same time. In detail, it targets the help to hyperactive children and their parents offered by schools and consequently the ratio of utilization by the parents. At the very end of the practical part of the diploma thesis, I dedicate my work to an assessment of hypothesis defined by myself.

Trichobilharzia regenti is a neurotropic fluke belonging to family Schistosomatidae. Larvae called schistosomula migrate in the definitive hosts (anseriform birds) throuth the central nervous system (CNS) to their final location in nasal mucosa, where they mature and lay eggs. In contrast with that, the infection of accidental mammalian hosts (including human), is often stopped already in the skin immediately after entering the host. However, some schistosomula are able to reach CNS of experimentally infected mice, and survive there temporarily. Reaction to the CNS infection of mice is usually provided by microglia, astrocytes or the other immune cells infiltrated from the hosts blood. Parasite protects itself against the host reaction with its tegument. It does not serve only as mechanical barrier, but also as main secretoric organ that is capable of active immune evasion. Changes within CNS of the vertebrate hosts, caused by migrating schistosomula of T. regenti, were already described by routine histological and immunohistochemical methods. Till now, there was a lack of informations about interactions of immune cells of the host and the tegument of the parasite on ultrastructural level. To fill this gap in knowledge, two different methods were used: (1) imunohistochemistry in light and electron...

Abstract Statement of problem Plasma angiotensin II (Ang II) increases blood pressure (BP) through the activation of brain angiotensinergic pathways and the aldosterone-mineralocorticoid receptors (MR)- epithelial Na+ channel (ENaC)-endogenous ouabain (EO) pathway. The response of BP to circulating Ang II is enhanced by high salt intake, but the central mechanisms mediating this elevated response are not known. Methods of investigation Study 1) Male Wistar rats were divided into 4 groups and treated with regular salt diet (0.4% NaCl), high salt diet (2% NaCl), sc Ang II infusion (150 ng/kg/min), or sc Ang II infusion together with 2% salt diet for 14 days; plasma aldosterone and corticosterone levels, CYP11B2 mRNA in adrenal cortex and the mRNA levels of Ang II type 1 receptors (AT1R), CYP11B1 (11-β hydroxylase), CYP11B2 (aldosterone synthase), MR, 11βHSD2, ENaC α, ENaC β and ENaC γ in the subfornical organ (SFO), paraventricular nucleus (PVN), supraoptic nucleus (SON) and rostral ventrolateral medulla (RVLM) were measured. Study 2) MR blockers (eplerenone, spironolactone), ENaC blocker (benzamil), AT1R blocker (losartan) or vehicles were centrally infused in rats treated with Ang II plus high salt, and BP and heart rate (HR) were recorded by telemetry; plasma aldosterone and corticosterone levels and CYP11B2 mRNA expression in adrenal cortex were measured. Major findings Ang II alone caused a small increase in BP. Ang II together with 2% salt diet markedly increased the BP and plasma aldosterone level. Sc Ang II decreased 11βHSD2 and MR mRNA expression in the PVN, increased AT1R and ENaC γ expression in the PVN, and increased AT1R mRNA expression in the RVLM. Other genes tested in the four brain nuclei were not affected by sc Ang II or high salt diet. BP and plasma aldosterone increases in response to Ang II and salt, as well as CYP11B2 mRNA expression in adrenal cortex, were largely prevented by central infusion of eplerenone, spironolactone, benzamil or losartan. Main conclusion BP and plasma aldosterone responses to Ang II-salt are under the control of central mechanisms, and MR-AT1R activation in the brain plays a critical role in Ang II-salt induced hypertension.

Encephalitis is a matter for serious public health concern because of the high morbidity and mortality associated with many cases. Epidemiological studies have shown that viral encephalitis (VE) is more common than the sum of encephalitis caused by all other pathogens. However, more than 95% of cases have no known cause. Thus, there is a significant need to develop a sensitive method for the diagnosis of these unknown cases. Previous sequence independent amplification (SIA) assays have proved successful in detecting new viruses in many biological samples but not in CSF samples. This may be due to the relatively low sensitivity of most available methods as CSF usually contains lower concentrations of pathogen than most other samples. A known problem with these types of assays is the annealing of the random primers to human DNA which facilitates preferential amplification of background human DNA. Thus, large scale sequencing is usually required to detect a virus, which in turn reduces the detection sensitivity to more than 1000 viral copies/µl, a CSF concentration that is rarely seen in cases of VE.This project was designed to develop a highly sensitive SIA assay for novel nucleic acid identification that could be used in testing CSF samples obtained from patients with neurological diseases of unknown cause. The study started with evaluation of two existing SIA assays commonly used for virus discovery; whole genome amplification (WGA) and random PCR (r-PCR). Sequential modification and adaptation of these methods was carried out to increase their sensitivity. Ultimately, a novel primer (Sa primer) that showed no binding to most human DNA sequences in GenBank was designed and synthesised. Its 3' end was tagged with 6 and 7 random nucleotides generating 2 r-primers; Sa-6 and Sa-7. The sensitivity of the r-primers was checked in a novel assay developed during this project and named Sa-SIA using known concentrations of HCMV and HSV-1. CSF samples from Malawian children were then tested using the developed assay. Results showed that adaptation of the existing WGA and r-PCR assays allowed detection of up to 1300 viral copies/µl. When the novel primers developed in this project were used in a random PCR assay (Sa-r-PCR), it was found that using Sa-6 primer 130, 13, and 1.3 HCMV copies/µl could be detected with 100, 60, and 50% efficiency respectively. When using Sa-7 primer, the same concentrations of virus were detected with 100, 42, and 28.6% efficiency. DNase-1 treatment of the samples pre-extraction resulted in an improvement in viral detection sensitivity in samples with a high background of host DNA. Starting with template concentrations of 11000, 110, 11, and 1.1 HSV-1 copies/µl, viral detection efficiency was increased from 33.3, 10, 0, and 0% to 92, 55.6, 16.7, and 0% respectively when pre-extraction DNase-1 treatment preceded Sa-r-PCR using Sa-6 primer. The final developed assay (Sa-SIA) consisted of centrifugation, DNase-1 treatment, DNA extraction, Sa-r-PCR using Sa-6 and Sa primers, gel electrophoresis, band excision, cloning, small scale sequencing (sequencing of ≤ 20 positive clones from one constructed DNA library), and bioinformatics. It had a detection sensitivity of 1.3-11 viral copies/µl. When this assay was applied to stored CSF samples, one 448bp sequence was identified which gave 96% coverage with 81% identity to Torque teno midi virus-1 and 93% coverage with 81% identity to small anellovirus-2. A 236bp sequence from another CSF sample showed 66% coverage with 97% homology to an unclassified sequence previously identified in a viral genomic survey of stool sample in an earlier published study. In conclusion, the standardised method had been shown to detect 1.3 to 11 viral copies/µl of two viruses; HCMV and HSV-1. The detection of these viruses was achieved with only small scale sequencing. Application of this method to CSF samples has shown promising results. However, this method could be followed by more advanced post amplification analyses such as next generation sequencing.

Numerous species of the genus Staphylococcus other than Staphylococcus aureus are important pathogens in human clinical practice and veterinary medicine. With improved methods of identification and more precise classification, we have speciated over 500 strains of staphylococci representing 17 species and subspecies of non-S. aureus Staphylococcus. We have examined these strains for possible virulence factors which may play a role in their pathogenesis. Using transmission electron microscopy (TEM), we have demonstrated that small to large amounts of glycocalyx are found on staphylococcal cells. Animal models have shown that staphylococci cause abscess formation in the presence or absence of a foreign body implant. Molecular characterization of cell extracts of Staphylococcus intermedius show that this species elaborates a protein which is serologically similar to the enterotoxins of Staphylococcus aureus in ELISA tests, but differs markedly in other characteristics.

Mucopolysaccharidosis (MPS) IIIB is a lysosomal storage disease with severe neurological manifestations due to a-N-acetylglucosaminidase (NaGlu) deficiency. The mechanism of neuropathology in MPS IIIB is unclear. This study investigates the role of immune responses in neurological disease of MPS IIIB in mice. By means of gene expression microarrays and realtime quantitative reverse transcriptase-polymerase chain reaction, we demonstrated significant up-regulation of numerous immune-related genes in MPS IIIB mouse brain involving a broad range of immune cells and molecules, including T cells, B cells, microglia/ macrophages, complement, major histocompatibility complex class I, immunoglobulin, Toll-like receptors, and molecules essential for antigen presentation. The significantly enlarged spleen and lymph nodes in MPS IIIB mice were due to an increase in splenocytes/lymphocytes, and functional assays indicated that the T cells were activated. An autoimmune component to the disease was further suggested by the presence of putative autoantigen or autoantigens in brain extracts that reacted specifically with serum IgG from MPS IIIB mice. We also demonstrated for the first time that immunosuppression with prednisolone alone can significantly slow the central nervous system disease progression. Our data indicate that immune responses contribute greatly to the neuropathology of MPS IIIB and should be considered as an adjunct treatment in future therapeutic developments for optimal therapeutic effect.

Increased survival of presynaptic-like neuronal profiles was found in cell cultures of rat cerebellum when the non-neuronal cell numbers were reduced with an antimitotic drug. In both treated and untreated cell cultures, neurites grew onto the polylysine-coated surface of sepharose beads and formed a swelling. The neuronal swelling contained an accumulation of synaptic vesicles and a membrane density at the site of contact with the bead and was called an apparent presynaptic element. The apparent presynaptic elements in untreated cultures increased in number from the time the beads were added to the culture to 7 days incubation and then showed a decrease to one half the 7-day value at 14 days incubation. A 75% reduction in cell division of non-neuronal cells was seen in cultures exposed to a 5 × 10-6 M cytosine arabinoside (Ara-C) for 2 days. Adding polylysine-coated beads to cultures treated with Ara-C showed at 14 days incubation a 7-fold increase in the number of apparent presynaptic elements as compared to untreated cultures. Additional experiments examined the numbers of neurites on the beads and found only small differences between treated and untreated cultures. A decrease, however, was shown in the number of glial fibrillary acidic protein staining astrocytes on the surface of the beads in treated cultures. The reduction of astrocytes by Ara-C appeared to enhance the survival of apparent presynaptic elements but did not enhance the growth of neurites. These results suggest that proliferating non-neuronal cells at a site of injury in the central nervous system may inhibit the formation of synaptic contacts and the growth of neurites through the site of injury.

Because endorphin receptor activation alters the function of the central noradrenergic system, opiates may change the regenerative sprouting of neurons in response to adrenergic neurotoxins. To test this hypothesis, newborn rats were treated with several opioids and 6-hydroxydopa (6-OHDOPA) and the development of the noradrenergic system was evaluated. In combination with 6-OHDOPA morphine and naloxone potentiated the development of norepinephrine (NE) levels in the pons-medulla and cerebellum by four weeks of age. β-Endorphin, Leu- and Met-enkephalin and d-Ala2-enkephalinamide produced a similar effect in the pons-medulla. The effect of morphine was partially attenuated by naloxone. Increased cerebellar noradrenergic histofluorescent staining was observed with the morphine + 6-OHDOPA and naloxone + 6-OHDOPA treatments. Both naloxone and morphine decreased NE levels in the pons-medulla of adult rats treated with 6-OHDOPA. These results suggest that opiates and endorphins may enhance sprouting of noradrenergic neurons following neonatal treatment with 6-OHDOPA, by increasing the toxicity of this neurotoxin.

The receptor tyrosine kinase, RON, is expressed on tissue-resident macrophages. RON functions by activating genes that promote wound repair and resolve inflammation, while repressing genes that perpetuate tissue damage and cell death. Chronic HIV infection is associated with dysregulated inflammation, and we hypothesize that diminished macrophage RON expression contributes to the development of end organ diseases including HIV-associated central nervous system (CNS) inflammation. We utilized CNS tissue from a SIV macaque model to examine the temporal regulation of RON in the brain during infection. Following prolonged SIV infection, RON expression was inversely correlated with the development of CNS disease: RON was highly expressed in animals that did not develop CNS lesions and lower in SIV infected macaques that demonstrated moderate to severe inflammatory lesions. Arginase-1 expression was low during late infection whereas expression of the inflammatory genes, IL-12 p40 and TNF &alpha, was elevated compared to uninfected animals. To validate a role for RON in regulating HIV, we infected human tonsillar tissue-resident macrophages. RON inhibited HIV replication in tissue-resident macrophages. Furthermore, HIV infection diminished RON in tonsil macrophages. We propose a model in which RON expression is decreased, genes that quell inflammation are repressed, and inflammatory mediators are induced to promote tissue inflammation following chronic HIV infection in the brain. The cyclin dependent kinase inhibitor p21 is a factor that, like RON, negatively regulates HIV transcription. Elevated expression of p21 in HIV+ elite controllers, or by ectopic expression in primary CD4+ T cells, resulted in reduced HIV expression. Furthermore, these elite controllers had increased binding of factors that negatively regulate transcription elongation at the HIV long terminal repeat. RON and p21 are examples of cellular factors that limit HIV transcription and contribute to HIV latency. Latently infected cells are not targeted by anti-retroviral therapy and permit rapid rebound of viremia following treatment interruption. Understanding intrinsic mechanisms that establish latency may provide targets for purging these HIV reservoirs or maintaining their transcriptionally silent state.