TOXOPLASMOSE NA GESTAÇÃO

A toxoplasmose, uma antropozoonose causada pelo protozoário Toxoplasma gondii, é uma infecção frequente em todo o mundo. Na maioria dos casos, não traz repercussões importantes para o paciente, exceto em indivíduos imunodeprimidos e fetos, os quais podem apresentar graves sequelas. A infecção materna geralmente resulta da ingestão de oocistos presentes no meio ambiente ou da ingestão de bradizoítos ou taquizoítos presentes em carnes ou produtos derivados. A transmissão fetal relaciona-se diretamente à idade gestacional na qual ocorreu a infecção materna, sendo maior o risco no terceiro trimestre. Testes sorológicos representam o método mais comum para se estabelecer o diagnóstico e o tratamento, que objetiva diminuir o risco de transmissão placentária do parasita, é empregado logo que é estabelecido o diagnóstico de infecção materna aguda. Palavras-chave: Toxoplamose. Gestação. Toxoplasma gondii

TOXOPLASMOSE NA GESTAÇÃO

A toxoplasmose, uma antropozoonose causada pelo protozoário Toxoplasma gondii, é uma infecção frequente em todo o mundo. Na maioria dos casos, não traz repercussões importantes para o paciente, exceto em indivíduos imunodeprimidos e fetos, os quais podem apresentar graves sequelas. A infecção materna geralmente resulta da ingestão de oocistos presentes no meio ambiente ou da ingestão de bradizoítos ou taquizoítos presentes em carnes ou produtos derivados. A transmissão fetal relaciona-se diretamente à idade gestacional na qual ocorreu a infecção materna, sendo maior o risco no terceiro trimestre. Testes sorológicos representam o método mais comum para se estabelecer o diagnóstico e o tratamento, que objetiva diminuir o risco de transmissão placentária do parasita, é empregado logo que é estabelecido o diagnóstico de infecção materna aguda. Palavras-chave: Toxoplamose. Gestação. Toxoplasma gondii

Integrated proteomics identified up-regulated focal adhesion-mediated proteins in human squamous cell carcinoma in an orthotopic murine model

Understanding the molecular mechanisms of oral carcinogenesis will yield important advances in diagnostics, prognostics, effective treatment, and outcome of oral cancer. Hence, in this study we have investigated the proteomic and peptidomic profiles by combining an orthotopic murine model of oral squamous cell carcinoma (OSCC), mass spectrometry-based proteomics and biological network analysis. Our results indicated the up-regulation of proteins involved in actin cytoskeleton organization and cell-cell junction assembly events and their expression was validated in human OSCC tissues. In addition, the functional relevance of talin-1 in OSCC adhesion, migration and invasion was demonstrated. Taken together, this study identified specific processes deregulated in oral cancer and provided novel refined OSCC-targeting molecules.Understanding the molecular mechanisms of oral carcinogenesis will yield important advances in diagnostics, prognostics, effective treatment, and outcome of oral cancer. Hence, in this study we have investigated the proteomic and peptidomic profiles by co95e98208FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO2009/54067-3; 2010/19278-0; 2011/22421-2; 2009/53839-2; 2011/02267-9470567/2009-0; 470549/2011-4; 301702/2011-0; 470268/2013-1; 505413/2013-

Modula??o do influxo de c?lcio pelo pept?deo YY (3-36) em c?lulas do hipocampo de ratos

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS ([email protected]) on 2015-08-27T11:15:23Z No. of bitstreams: 1 474252 - Texto Completo.pdf: 3311481 bytes, checksum: b67f5f48a8325dd3ae5469933fff90ba (MD5)Made available in DSpace on 2015-08-27T11:15:23Z (GMT). No. of bitstreams: 1 474252 - Texto Completo.pdf: 3311481 bytes, checksum: b67f5f48a8325dd3ae5469933fff90ba (MD5) Previous issue date: 2015-03-02Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPESConselho Nacional de Pesquisa e Desenvolvimento Cient?fico e Tecnol?gico - CNPqPeptide YY (PYY) belongs to the neuropeptide Y (NPY) family, which also includes the neuropeptide Y (NPY) and pancreatic polipeptide (PP). These substances are biologically active, constituted of 36 aminoacids, and act via G protein coupled receptors. There are four functional subtypes of NPY family receptors in humans, namely Y1, Y2, Y4, and Y5. PYY is secreted by the intestinal L cells, being present in the blood stream in two active forms capable of crossing the blood brain barrier, PYY (1-36) and its cleavage product, PYY (3-36). PYY is a selective agonist for the Y2 receptor (Y2R) and has been identified as a modulator of appetite, promoting satiety sensation in mammals. Y2R are abundant in the brain hippocampus, where these receptors inhibit excitatory synaptic transmission and glutamatergic release when activated by potassium in hippocampal slices. Besides, knockout mice for Y2R present deficits in spatial and non-spatial memory tasks, showing a role of Y2R in learning and memory. The aim of this Master?s dissertation was searching for a better understanding of the interaction of PYY (3-36) and its Y2 receptor in CNS cells. For this purpose the activity of this peptide was investigated on Ca2+ influx in hippocampal cell cultures of Wistar neonate rats. We evaluated the influence of the presence of Ca2+ in the extracellular fluid, as well as the involvement of plasma membrane voltage-dependent Ca2+ and Na+ channels, and the influence of intracellular mechanisms related to the endoplasmic reticulum (ER), such as the SERCA pump, the inositol triphosphate (IP3) receptors and the ryanodine receptors (RyRs) in the responses induced by PYY (3-36) on the modulation of the [Ca2+]i, by using blockers specific for these channels. It was observed that the increase of the cytosolic [Ca2+] evoked by PYY (3-36) in hippocampal cells is independent of Ca2+ from the extracellular environment. Using a voltage-dependent Na+ channels blocker it was possible demonstrate that PYY (3-36) action is independent on Na+, suggesting that its activity on hippocampal cells does not induce or does not depend on cellular depolarization. In the experiments using RyRs or SERCA pump blockers it was observed an elevation of Ca2+ influx, that probably occurred due to the activation of SOCC, but with the concomitant presence of the a voltage-dependent Na+ channels blocker, this effect was abolished, suggesting a probable inhibition of SOCC channels in these conditions. In the experiments in the presence of an IP3Rs inhibitor, there was a decrease in cytosolic [Ca2+] evoked by PYY (3-36). The results from our experiments indicate that the action of PYY (3-36) on Ca2+ mobilization is mediated by intracellular receptors of the ER, suggesting that the observed elevation of [Ca2+]i is modulated especially by the activation of the intracellular Ca2+ signalling cascade through IP3 receptors.O pept?deo YY (PYY) pertence ? fam?lia do neuropept?dio Y que ? composta tamb?m pelo neuropept?dio Y (NPY) e pelo polipept?dio pancre?tico (PP). S?o pept?deos biologicamente ativos, compostos por 36 amino?cidos e atuam via receptores acoplados a prote?na G. Existem quatro subtipos de receptores para a fam?lia do NPY que s?o funcionais em humanos, denominados como: Y1, Y2, Y4 e Y5. O PYY ? secretado no intestino pelas c?lulas L e circula no organismo em duas formas ativas que atravessam a barreira hematoencef?lica, que s?o o PYY (1-36) e sua forma clivada PYY (3-36). O PYY (3-36) ? um agonista seletivo do receptor Y2 e tem sido evidenciado por seu papel como um modulador do apetite, promovendo a sensa??o de saciedade em mam?feros. O hipocampo ? uma regi?o rica em receptores Y2 e j? se demonstrou que a ativa??o destes receptores no hipocampo pode inibir a transmiss?o sin?ptica excitat?ria e a libera??o de glutamato estimulada por pot?ssio em fatias hipocampais. Em camundongos knockout para o receptor Y2 observou-se d?ficits na mem?ria espacial e na mem?ria n?o espacial nestes animais evidenciando tamb?m seu envolvimento na regula??o da fun??o cognitiva associada com o aprendizado e mem?ria. O desenvolvimento desta disserta??o teve por objetivo buscar uma melhor compreens?o da intera??o do PYY (3-36) e seu receptor Y2 em c?lulas do sistema nervoso central. Para isso investigou-se a atividade deste pept?deo no influxo de c?lcio (Ca2+) em c?lulas cultivadas do hipocampo de ratos Wistar neonatos. Avaliou-se a influ?ncia do Ca2+ presente no meio extracelular, bem como o envolvimento dos canais de Ca2+ e Na+ voltagem dependentes presentes na membrana plasm?tica, e a influ?ncia de mecanismos intracelulares presentes no ret?culo endoplasm?tico (RE), como a bomba do ret?culo sarco/endoplasm?tico Ca2+- ATPase (SERCA), os receptores de inositol trifofato (IP3Rs) e os receptores de rianodina (RyRs) nas respostas induzidas pelo PYY (3-36) sobre a modula??o da concentra??o de Ca2+ intracelular ([Ca2+]i), atrav?s da utiliza??o de bloqueadores espec?ficos para estes canais. Foi observado que o aumento da concentra??o de Ca2+ ([Ca2+]) citos?lico evocado pelo PYY (3-36) em c?lulas do hipocampo ? independente do Ca2+ do meio extracelular. E atrav?s da utiliza??o de bloqueadores dos canais de Na+ voltagem dependentes conseguiu-se demonstrar que a a??o do PYY (3-36) ? independente do influxo de Na+, sugerindo que a sua atividade sobre as c?lulas hipocampais n?o induz ou independe da despolariza??o celular. Nos experimentos utilizando bloqueadores dos RyRs ou da bomba SERCA observou-se uma eleva??o do influxo de Ca2+, o que provavelmente ocorreu devido a ativa??o dos canais de c?lcio operados por estoque (SOCC, do ingl?s store-operated calcium channels), por?m na presen?a concomitante de bloqueadores de canais de Na+ voltagem dependentes, este efeito foi bloqueado, sugerindo uma prov?vel inibi??o dos canais SOCC nessas condi??es. Finalmente, na presen?a de inibidores dos IP3Rs, ocorreu uma diminui??o da [Ca2+] citos?lico evocada pelo PYY (3-36) nas c?lulas. Os resultados evidenciam que a a??o do PYY (3-36) sobre a mobiliza??o do Ca2+ ? atrav?s de receptores intracelulares do RE, sugerindo que a eleva??o da [Ca2+]i observada, ? modulada principalmente pela ativa??o da cascata de sinaliza??o do Ca2+ intracelular pelos receptores de IP3

Integrated Proteomics Identified Up-Regulated Focal Adhesion-Mediated Proteins in Human Squamous Cell Carcinoma in an Orthotopic Murine Model

<div><p>Understanding the molecular mechanisms of oral carcinogenesis will yield important advances in diagnostics, prognostics, effective treatment, and outcome of oral cancer. Hence, in this study we have investigated the proteomic and peptidomic profiles by combining an orthotopic murine model of oral squamous cell carcinoma (OSCC), mass spectrometry-based proteomics and biological network analysis. Our results indicated the up-regulation of proteins involved in actin cytoskeleton organization and cell-cell junction assembly events and their expression was validated in human OSCC tissues. In addition, the functional relevance of talin-1 in OSCC adhesion, migration and invasion was demonstrated. Taken together, this study identified specific processes deregulated in oral cancer and provided novel refined OSCC-targeting molecules.</p></div

Talin-1 knockdown decreased cell adhesion, migration and invasion of SCC-9, A431 and SCC-9 LN1 cells.

<p>(a) Talin-1 showed lower protein expression in SCC-9/siRNA TLN-1 cells compared to SCC-9/control (scrambled) by immunoblotting. The total proteins (30 µg) were submitted to 1-D electrophoresis on 12% SDS-polyacrylamide gels, they were transferred onto nitrocellulose membrane and incubated with anti-talin-1 antibody. Anti-GAPDH antibody was used as loading control. The graph represents the normalized optical density. (b) Talin-1 mRNA expression levels in SCC-9/siRNA TLN-1 cells compared to SCC-9/control (scrambled) by qRT-PCR (n = 3, Student's <i>t</i>-test, p<0.05). The data were normalized with GAPDH gene. (c) SCC-9/control (scrambled) and SCC-9/siRNA TLN-1 cells, A431/control (scrambled) and A431/siRNA TLN-1 cells, SCC-9 LN1/control (scrambled) and SCC-9 LN1/siRNA TLN-1 cells were seeded in Matrigel coated 96-well plates. After 1 h, cells were stained and the cell adhesion was measured (<b>n = 3</b>, * indicates p<0.05, Student's <i>t</i>-test for each comparison). (d) SCC-9/control (scrambled) and SCC-9/siRNA TLN-1 cells, A431/control (scrambled) and A431/siRNA TLN-1 cells, SCC-9 LN1/control (scrambled) and SCC-9 LN1/siRNA TLN-1 cells were seeded in serum-free media in the upper chamber of transwell plates and were allowed to migrate towards the lower chamber containing 1% FBS supplemented media (n = 3, * indicates p<0.05, Student's <i>t</i>-test). (e) SCC-9/control (scrambled) and SCC-9/siRNA TLN-1 cells, A431/control (scrambled) and A431/siRNA TLN-1 cells, SCC-9 LN1/control (scrambled) and SCC-9 LN1/siRNA TLN-1 cells were seeded in serum-free media in the upper chamber of matrigel-coated transwell plates and were allowed to invade towards the lower chamber containing 10% FBS supplemented media (n = 2, * indicates p<0.05, Student's <i>t</i>-test).</p