Avaliação da técnica imunohistoquímica como método de deteção da apoptose a nível celular

A apoptose constitui um processo fisiológico de morte celular, caracterizado por alterações morfológicas distintas e mecanismos bioquímicos e moleculares bem definidos. O seu papel de destaque em numerosos eventos biológicos e importantes processos patológicos conduziu a um crescente interesse na investigação dos mecanismos celulares que regulam o processo apoptótico. A aplicação de metodologias capazes de identificar células apoptóticas despoletou um enorme desenvolvimento de técnicas. No entanto, as propriedades demonstradas por estes ensaios nem sempre se aplicam ao estudo de amostras tecidulares, pelo que a escolha dos diferentes métodos deverá ser criteriosamente avaliada, tendo em conta a aplicação pretendida e as alterações morfológicas que se pretendem detetar. Das várias técnicas disponíveis para deteção da apoptose em tecidos, muitos investigadores recomendam o método TUNEL, o qual se baseia na marcação de produtos endonucleossómicos resultantes da fragmentação do DNA. Outros métodos histoquímicos também disponíveis incluem a deteção do citocromo c, libertado da mitocôndria ou a deteção das proteínas pró e anti-apoptóticas, Bax, Bidm e Bcl-2, envolvidas nos mecanismos intrínsecos da apoptose. Mais recentemente, a marcação de produtos específicos resultantes da clivagem de proteínas alvo pelas caspases, tem vindo a ser considerada uma abordagem promissora. Como principal objectivo deste trabalho pretendeu-se avaliar a técnica imunohistoquímica como método de deteção da apoptose a nível celular, em tecidos animais, tendo por base o método TUNEL, o qual permite a deteção de fragmentos de DNA. Os resultados obtidos permitiram concluir que, apesar do método TUNEL possuir as suas limitações ao nível da sensibilidade e especificidade, o mesmo constitui um mecanismo imunohistoquímico útil na deteção de células apoptóticas. Contudo, segundo opinião de vários autores, adverte-se para a necessidade da aplicação de pelo menos dois métodos imunohistoquímicos como forma de validar a ocorrência do processo apoptótico, razão pela qual se optou pela deteção do citocromo c citosólico como método complementar, uma vez que a sua libertação para o espaço citosólico se encontra implicada na ativação da apoptose.

O papel do médico dentista no diagnóstico precoce de tumores malignos do seio maxilar

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

La patología celular : basada en el estudio fisiológico y patológico de los tejidos /

Mode of access: Internet.

Sistema penitenciario : el presidio, la prisión celular, la deportación /

Mode of access: Internet.

FATORES DETERMINANTES DA SOBREVIVÊNCIA DAS MICRO E PEQUENAS EMPRESAS DO SETOR INDUSTRIAL NA CIDADE DE SANTOS

A importância das micro e pequenas empresas na economia brasileira já é sentida desde há muito tempo, pois esse segmento é fundamental na geração de riqueza para a sociedade. Apesar da importância econômica, ainda persiste um elevado grau de descontinuidade nas mesmas, o que lhes outorga dificuldades de transporem os dois primeiros e mais críticos anos de existência. Esta pesquisa visa identificar os fatores determinantes da sobrevivência das micro e pequenas empresas do setor industrial na cidade de Santos. A relevância do estudo se intensifica em decorrência da cidade de Santos estar vivenciando um momento de desenvolvimento econômico impulsionado pelas descobertas de petróleo na camada do pré-sal. Isso poderá aumentar ainda mais o número de criação de MPEs, principalmente no setor industrial, e que provavelmente necessitarão de informações nas quais possam se espelhar. O estudo é de caráter exploratório e descritivo. Os dados foram colhidos por meio de uma pesquisa de campo com aplicação de um questionário baseado em quatro dimensões: empreendedorismo, ambiental, recursos financeiros e organização interna. A pesquisa apresentou como principais fatores de sobrevivência o tempo dedicado à empresa; a adequação dos produtos a demanda; a capacidade de assumir riscos e a liderança do gestor. A amostra de estudo foi definida por conveniência, constituída de MPEs pertencentes ao setor econômico industrial, pois é o mais aderente à cadeia de exploração de petróleo. Foram consideradas as empresas que ultrapassaram os dois primeiros anos de existência, de acordo com os critérios de avaliação de sobrevivência adotado pelo SEBRAE (2011).

A ANÁLISE DAS CAPACIDADES DE SOBREVIVÊNCIA DE PEQUENOS NEGÓCIOS NAS CIDADES DE ITANHAÉM E MONGAGUÁ: ESTUDO MULTICASO DE PEQUENAS EMPRESAS

O presente trabalho traz uma análise das capacidades de sobrevivência de três micro e pequenas empresas (MPEs) do ramo alimentício de doces de banana, estabelecidas na Região Metropolitana da Baixada Santista, especificamente nas cidades de Itanhaém e Mongaguá. Essas capacidades se construíram fundamentadas em dois pressupostos: efeito do gestor e o efeito da localidade, que podem ser determinantes para a sobrevivência e o desempenho de MPEs. A pesquisa tem por objetivo geral identificar as características do gestor e os fatores da localidade que influenciam a sobrevivência das três MPEs estudadas. Para alcançar o objetivo geral foram traçados os seguintes objetivos específicos: identificar as características da gestão que foram propulsoras da sobrevivência das três MPEs estudadas; identificar os recursos da localidade que foram propulsores à sobrevivência das três MPEs; verificar a existência de outros fatores da gestão e da localidade que possam ter influenciado a sobrevivência das MPEs no longo prazo. O estudo teve como expectativa contribuir para o aprofundamento da reflexão sobre o tema concernente às capacidades de sobrevivência dos pequenos negócios. A pesquisa bibliográfica esteve centrada no encadeamento de tópicos teóricos de forma a construir fundamentos necessários sobre as influências do gestor e da localidade na sobrevivência das pequenas empresas, noções necessárias para se atingir o objetivo geral. Para alcançar os objetivos propostos, a investigação, de caráter descritivo e indutivo, foi desenvolvida por meio de estudo multicaso, compreendendo pesquisa documental, observações e entrevistas semi-estruturadas, preliminar e de profundidade, tendo como sujeitos os gestores, os funcionários, os fornecedores e os clientes das três MPEs. Os resultados indicam que há fatores importantes do efeito gestor e do efeito localidade influenciando a sobrevivência das empresas locus da pesquisa. Os resultados apontaram fatores como acesso ao amplo mercado, atitude frente à tecnologia, a presença de laço forte e laço fraco, entre outros considerados importantes influenciadores na sobrevivência das empresas estudadas. Os resultados abrem perspectivas de estudos mais aprofundados que possam proporcionar e criar alternativas e programas para melhorar o desempenho e a sobrevivência das MPEs.(AU)

TELEFONE CELULAR: A nova mídia de massa a serviço da publicidade segmentada.

Ao final do ano de 2004 o Brasil atingiu o número de mais de 50 milhões de aparelhos celulares, ultrapassando inclusive o número de telefones fixos. No mesmo período, as operadoras de telefonia móvel lançaram o sinal de televisão para serem recebidos nestes aparelhos, que são pessoais e portáteis. O objetivo deste trabalho é verificar se em virtude deste crescimento na base de usuários e da alta tecnologia o aparelho de telefone celular tem potencial como mídia publicitária, sendo capaz de segmentar suas mensagens de acordo com o perfil dos clientes.

Os efeitos das queimadas na Amazônia em nível celular e molecular

The Amazon holds over half of the planet's remaining tropical forests and comprises the largest biodiversity in the world, accounting for approximately 60 % of the Brazilian territory. However, deforestation fires in the region causes serious problems to exposed human. The aim of this study was to evaluate the chemical compounds as well as the cellular and molecular effects after exposure to organic material extracted from particulate matter less than 10 µm (PM10) in the Amazon region. As for the chemical composition, n-alkanes analysis showed a prevalence of anthropogenic influence during the fires in the region. In addition, there was a predominance of monosaccharides from biomass burning markers. Also, the Polycyclic Aromatic Hydrocarbons (PAH) and their derivatives have also been identified in samples collected in the Amazon. By using the PAH concentrations was possible to calculate the BaP-equivalent and it was found that the dibenz(a) anthracene contributes with 83% to potential carcinogenic risk. As for the potential mutagenic risk, the benzo (a) pyrene is the HPA that has a major contribution in this analysis. It may be noted that the retene was the most abundant PAH. This compound was genotoxic and cause death by necrosis in the human lung cells. In biological tests, the data showed that organic PM10 is capable of causing genetic damage in both plant cells and in human lung cells. This damage cause an arrest in the G1 phase of the cell cycle exposed, increasing the expression of p53 and p21. Additionally, the PM10 caused cell death by apoptosis, increasing the foci of histone - H2AX. Given these results, it is important to emphasize the reduction and better control of biomass burning in the Amazon region thus improving the quality of health of the population being exposed. As clearly stated recently by the World Health Organization, the reduction of air pollution could save millions of lives annually.

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.

Moléculas coestimulatórias na leishmaniose visceral

Visceral leishmaniasis (VL) is endemic in many countries, including Brazil. The protozoan Leishmania infantum, is the etiological agent of VL, and is transmitted by the bite of female sandflies during the blood meal. The majority of subjects when exposed to the parasite do not develop the disease, because of development of Th1 cellular responses. Those who have develop signs of VL such as fever, weight loss, hepatosplenomegaly, have impairment of the cellular immune response, specific to the Leishmania antigens. We evaluated whether the specififc anergy during symptomatic VL, may be associated with changes in T cells costimulatory molecules or their ligands in CD14+ monocytes. There is an increase in CTLA-4 porcentage on CD4+ T lymphocytes (p=0.001) and ICOS on CD4+ and CD8+ T cells (p=0.002 to CD4+ and p=0.003 to CD8+), after stimulation by soluble Leishmania antigen (SLA) during active visceral leishmaniasis, and that there is a higher percentage of these molecules ex vivo, when comparing symptomatic to recovered individuals (p=0.04 to CTLA-4 in CD4+, and p=0.001 to ICOS in CD4+ and p=0.026 to CD8+). Moreover, we found a high gene expression of CTLA-4, OX-40 and ICOS during active VL. CD40, CD80, CD86, HLA-DR and ICOSL molecules do not suffer changes during disease. There is IFN-γ production by the peripheral blood cells, after SLA stimulation, by peripheral blood cells in symptomatic subjects; however, there is a decrease of the ratio IFN-γ/IL-10, which is reversed after clinical recovery. The impairment of some costimulatory molecules pathways during symptomatic VL could inhibit the ability of phagocytes to kill Leishmania and could facilitate their survival and the proliferation inside macrophages.

Avaliação do efeito da inibição do reparo de sítios abásicos na resposta inflamatória celular

Base excision repair (BER) proteins has been associated with functions beyond DNA repair. Apurynic/apyrimidinic endonuclease 1 (APE1) is a multifunctional protein involved in a plethora of cellular activities, such as redox activation of transcription factors, RNA processing and DNA repair. Some studies have described the action of the protein 8-oxoguanine (OGG1) in correcting oxidized lesions in promoters as a step in the transcription of pro-inflammatory cytokines. Despite being especially important in redox activation of transcription factors such as nuclear factor κB (NF-κB) and AP- 1, the repair activity of APE1 has not yet been associated with the inflammatory response. In this study, experimental and bioinformatic analysis approaches have been used to investigate the relationship between inhibition of the repair of abasic sites in DNA by MX, a synthetic molecule designed to inhibt the repair activity of APE1, and the modulation of the inflammatory response. The results showed that treatment of monocytes with lipopolysaccharide (LPS) and MX reduced the expression of cytokines, chemokines and toll-like receptors, and negatively regulated biological immune processes, as macrophages activation, and NF-κB and tumor necrosis factor (TNF-α) and interferon pathways, without inducing cell death. The transcriptomic analysis suggests that LPS/MX treatment induces mitochondrial dysfunction, endoplasmic reticulum stress and activation of autophagy pathways, probably activated by impairment of cellular energy and/or the accumulation of nuclear and mitochondria DNA damage. Additionally, it is proposed that the repair activity of APE1 is required for transcription of inflammatory genes by interaction with abasic sites at specific promoters and recruitment of transcriptional complexes during inflammatory signaling. This work presents a new perspective on the interactions between the BER activity and the modulation of inflammatory response, and suggests a new activity for APE1 protein as modulator of the immune response in a redox-independent manner.

“Acham que brotamos das fontes dessa cidade?”: uma etnografia sobre o cotidiano de sobrevivência de pessoas em situação de rua em Nata/RN

Through participant observation, held in the administrative region east of Natal / RN, this research aimed to understand about the daily life of men and women living on the streets in the city. How they relate to the space where they live? In this relationship, which uses and survival strategies triggered by this social segment? These were some of the questions that guided this research, in order to highlight the specificities of this way of life and the possible consequences that such a situation could reverberate. In this sense, there was an effort to keep up with people on the streets, their itineraries and seize their narratives. Along the search path - which took place intermittently between the years 2011-2015 - attended spaces of occupation and traffic of this population group in the street, as well as insert me and got involved in events, forums, seminars, meetings and subject of joints on the streets as a political movement (MNPR / RN). They are considered in this study as people on the street to those who occupy the street as their main space of survival and ordering of daily life: in the streets sleep, feed themselves, meet the physiological and hygienic needs, and is where draw sustenance. The street is taken in this research in its broadest sense, including all possible places relatively protected from the cold, rain and exposure to violence, thus includes both open and public spaces: as squares and parks; but also closed and private places: hostels, abandoned warehouses, prisons, etc. It was observed that in none of these spaces guys on the street are established in a fixed manner, in contrast, they experience the roaming, which in part is due to urban systems - which tends to stigmatize them and delete them places - and the very need to survive, because while living in the street differentiated practices are triggered, and these differ from the dominant mode of sedentary life.

“Acham que brotamos das fontes dessa cidade?”: uma etnografia sobre o cotidiano de sobrevivência de pessoas em situação de rua em Nata/RN

Through participant observation, held in the administrative region east of Natal / RN, this research aimed to understand about the daily life of men and women living on the streets in the city. How they relate to the space where they live? In this relationship, which uses and survival strategies triggered by this social segment? These were some of the questions that guided this research, in order to highlight the specificities of this way of life and the possible consequences that such a situation could reverberate. In this sense, there was an effort to keep up with people on the streets, their itineraries and seize their narratives. Along the search path - which took place intermittently between the years 2011-2015 - attended spaces of occupation and traffic of this population group in the street, as well as insert me and got involved in events, forums, seminars, meetings and subject of joints on the streets as a political movement (MNPR / RN). They are considered in this study as people on the street to those who occupy the street as their main space of survival and ordering of daily life: in the streets sleep, feed themselves, meet the physiological and hygienic needs, and is where draw sustenance. The street is taken in this research in its broadest sense, including all possible places relatively protected from the cold, rain and exposure to violence, thus includes both open and public spaces: as squares and parks; but also closed and private places: hostels, abandoned warehouses, prisons, etc. It was observed that in none of these spaces guys on the street are established in a fixed manner, in contrast, they experience the roaming, which in part is due to urban systems - which tends to stigmatize them and delete them places - and the very need to survive, because while living in the street differentiated practices are triggered, and these differ from the dominant mode of sedentary life.

Expressão de MMPs, marcadores angiogênicos e proliferação celular em tumores odontogênicos

INTRODUCTION AND OBJECTIVE: The study of biological behavior of odontogenic lesions is essential to the establishment of appropriate therapeutic approach and prognosis. The production of extracellular matrix metalloproteinases (MMPs), angiogenesis and cell proliferation contribute to tumor growth. This paper aims to review the literature on odontogenic tumors (OT) selected according to the new World Health Organization classification (WHO- 2005) by evaluating the expression of MMPs, angiogenic and cell proliferation. Furthermore, it aims to verify the relation between these markers and the biological behavior of these lesions. RESULTS: it was found that MMPs -1, -2, -7, -9 and -26 had a higher expression in both epithelial component and stroma, and 13 particularly in the stroma. Increased angiogenesis was observed in more aggressive OT. CD105 expression was higher in keratocystic odontogenic tumour (KOT) and CD34 in solid ameloblastomas (SA). It was observed a higher expression of Ki-67 and p53 in SA and KOT and a low cell proliferation rate in the adenomatoid odontogenic tumour (AOT). CONCLUSION: These results show that MMPs are involved in invasion and recurrence of some odontogenic lesions and are associated with the biological behavior of these tumors. Angiogenesis is critical to provide support to cell proliferation and these concomitant events are correlated with different levels of biological behavior in OT when compared to odontogenic cysts, hence the use of angiogenic inhibitors may be a potential therapeutic approach in these lesions.