28 resultados para Percolation by invasion

em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"


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Paracoccidioides brasiliensis probably uses many different mechanisms to establish itself in the host and cause disease. In this work, we assess an in vitro model system which uses cultured mammalian cells to investigate the virulence factors of P. brasiliensis. We were able to demonstrate an invasion process of the yeast form of this fungus in Vero cell cultures. We deduced that the overall invasive process involved three steps: adhesion, followed by invasion of individual epithelial cells and spread to adjacent cells.

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The Marsh Antwren (Stymphalornis acutirostris) is restricted to the lowlands between Antonina Bay, in the coastal plain of the state of Paraná, and Itapocu river, in the northern coastal plain of the state of Santa Catarina (from 0 to c. 5 m a.s.l.). It doesn't occur continuously in this region, being found in eight populations that span over an total area of about 6,060 ha (= area of occupancy; 4,856.67 in Paraná and c. 1,200 in Santa Catarina). Nine habitat types used by the Marsh Antwren were defined, based on vegetation physiognomy, localization, dominancy of botanical species, dominant life-form and history of the region. Five of these are herbaceous (marshes), while four have an upper arboreal stratum and an herbaceous lower stratum with marsh plants. According to the classification criteria of the Brazilian vegetation proposed by the Radambrasil Project, they were classified as Pioneering Formation of Fluvial Influence, Pioneering Formation of Fluvial-marine Influence, and/or Pioneering Formation of Lacustrine Influence. They occur as patches or narrow strips ranging from 0.001 to 203.0 ha in the state of Paraná. They are found mainly in the interior of bays, in the lower courses of rivers that drain into bays, in alluvial plains, and between sand dunes in the coastal plain. Characteristic herbaceous species are cattail (Typha domingensis), bulrush (Scirpus californicus), Crinum salsum, Panicum sp. cf. P. mertensii, saw grass (Cladium mariscus) and Fuirena spp. Hibiscus pernambucensis is the characteristic bush species, and Calophyllum brasiliense, Tabebuia cassinoides, Annona glabra and Laguncularia racemosa are the characteristic arboreal species. The Marsh Antwren lives in herbaceous vegetation, but also uses bushes and branches of small tress. It has low flight capacity and a single flight of more than 25 m was never recorded. Territories of 0.25 ha were estimated in one kind of habitat (tidal marsh) (= 8 individuals per hectare) and of 3.2 ha in another one (saw grass marsh) (= 0.62 individual per hectare). The global population estimate is of about 17,700 mature individuals (13,700 in Paraná and 4,000 in Santa Catarina). The species is really under threat of extinction, mainly because of it's restricted geographical distribution and habitat loss by human activities and biological contamination caused by invasion of exotic grasses (Urochloa arrecta and Brachiaria mutica).

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Paracoccidioides brasiliensis (Pb) yeast cells can enter mammalian cells and probably manipulate the host cell environment to favor their own growth and survival. We studied the uptake of strain Pb 18 into A549 lung and Vero epithelial cells, with an emphasis on the repercussions in the cytoskeleton and the apoptosis of host cells. Cytoskeleton components of the host cells, such as actin and tubulin, were involved in the P. brasiliensis invasion process. Cytochalasin D and colchicine treatment substantially reduced invasion, indicating the functional participation of microfilaments (MFs) and microtubules (MTs) in this mechanism. Cytokeratin could also play a role in the P. brasiliensis interaction with the host. Gp43 was recognized by anti-actin and anti-cytokeratin antibodies, but not by anti-tubulin. The apoptosis induced by this fungus in infected epithelial cells was demonstrated by various techniques: TUNEL, DNA fragmentation and Bak and Bcl-2 immunocytochemical expression. DNA fragmentation was observed in infected cells but not in uninfected ones, by both TUNEL and gel electrophoresis methods. Moreover, Bcl-2 and Bak did not show any differences until 24 h after infection of cells, suggesting a competitive mechanism that allows persistence of infection. Overexpression of Bak was observed after 48 h, indicating the loss of competition between death and survival signals. In conclusion, the mechanisms of invasion of host cells, persistence within them, and the subsequent induction of apoptosis of such cells may explain the efficient dissemination of P. brasiliensis. (C) 2004 Published by Elsevier SAS.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The 5-year survival rate for oral cavity cancer is poorer than for breast, colon or prostate cancer, and has improved only slightly in the last three decades. Hence, new therapeutic strategies are urgently needed. Here we demonstrate by tissue micro array analysis for the first time that RNA-binding protein La is significantly overexpressed in oral squamous cell carcinoma (SCC). Within this study we therefore addressed the question whether siRNA-mediated depletion of the La protein may interfere with known tumor-promoting characteristics of head and neck SCC cells. Our studies demonstrate that the La protein promotes cell proliferation, migration and invasion of lymph node-metastasized hypopharyngeal SCC cells. We also reveal that La is required for the expression of beta-catenin as well as matrix metalloproteinase type 2 (MMP-2) within these cells. Taken together these data suggest a so far unknown function of the RNA-binding protein La in promoting tumor progression of head and neck SCC.

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Background: Suppressor of cytokine signaling 3 (SOCS3) is an inducible endogenous negative regulator of signal transduction and activator of transcription 3 (STAT3). Epigenetic silencing of SOCS3 has been shown in head and neck squamous cell carcinoma (HNSCC), which is associated with increased activation of STAT3. There is scarce information on the functional role of the reduction of SOCS3 expression and no information on altered subcellular localization of SOCS3 in HNSCC.Methodology/Principal Findings: We assessed endogenous SOCS3 expression in different HNSCC cell lines by RT-qPCR and western blot. Immunofluorescence and western blot were used to study the subcellular localization of endogenous SOCS3 induced by IL-6. Overexpression of SOCS3 by CMV-driven plasmids and siRNA-mediated inhibition of endogenous SOCS3 were used to verify the role of SOCS3 on tumor cell proliferation, viability, invasion and migration in vitro. In vivo relevance of SOCS3 expression in HNSCC was studied by quantitative immunohistochemistry of commercially-available tissue microarrays. Endogenous expression of SOCS3 was heterogeneous in four HNSCC cell lines and surprisingly preserved in most of these cell lines. Subcellular localization of endogenous SOCS3 in the HNSCC cell lines was predominantly nuclear as opposed to cytoplasmic in non-neoplasic epithelial cells. Overexpression of SOCS3 produced a relative increase of the protein in the cytoplasmic compartment and significantly inhibited proliferation, migration and invasion, whereas inhibition of endogenous nuclear SOCS3 did not affect these events. Analysis of tissue microarrays indicated that loss of SOCS3 is an early event in HNSCC and was correlated with tumor size and histological grade of dysplasia, but a considerable proportion of cases presented detectable expression of SOCS3.Conclusion: Our data support a role for SOCS3 as a tumor suppressor gene in HNSCC with relevance on proliferation and invasion processes and suggests that abnormal subcellular localization impairs SOCS3 function in HNSCC cells.

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Marine biological invasions have been regarded as one of the major causes of native biodiversity loss, with shipping and aquaculture being the leading contributors for the introductions of alien species in aquatic ecosystems. In the present study, five aquatic alien species (one mollusk, three crustaceans and one fish species) were detected during dives, shore searches and from the fisheries on the coast of the Delta do Parnaiba Environmental Protection Area, in the States of Piaui and Maranhao, Northeastern Brazil. The species were the bicolor purse-oyster Isognomon bicolor, the whiteleg shrimp Litopenaeus vannamei, the giant river prawn Macrobrachium rosenbergii, the Indo-Pacific swimming crab Charybdis hellerii and, the muzzled blenny Omobranchus punctatus. Ballast water (I. bicolor, C. hellerii, and O. punctatus) and aquaculture activities (L. vannamei and M. rosenbergii) in adjacent areas are the most likely vectors of introduction. All exotic species found have potential impact risks to the environment because they are able to compete against native species for resources (food and habitat). Isognomon bicolor share the same habitat and food items with the native bivalve species of mussels and barnacles. Litopenaeus vannamei share the same habitat and food items with the native penaeids such as the pinkspot shrimp Farfantepenaeus brasiliensis, the Southern brown shrimp Farfantepenaeus subtilis, and the Southern white shrimp Litopenaeus schmitti, and in the past few years L. vannamei was responsible for a viral epidemics in the cultivation tanks that could be transmitted to native penaeid shrimps. Charybdis hellerii is also able to cause impacts on the local fisheries as the species can decrease the populations of native portunid crabs which are commercialized in the studied region. Macrobrachium rosenbergii may be sharing natural resources with the Amazon River prawn Macrobrachium amazonicum. Omobranchus punctatus shares habit with the native redlip blenny Ophioblennius atlanticus and other fishes, such as the frillfin goby Bathigobius soporator. Some immediate remedial measures to prevent further introductions from ballast water and shrimp farm ponds should be: (i) to prevent the release of ballast water by ship/vessels in the region; (ii) to reroute all effluent waters from shrimp rearing facilities through an underground or above-ground dry well; (iii) to install adequate sand and gravel filter which will allow passage of water but not livestock; (iv) outdoor shrimp pounds located on floodable land should be diked, and; (v) to promote environmental awareness of those directly involved with ballast water (crews of ship/vessels) and shrimp farms in the region. Rev. Biol. Trop. 58 (3): 909-923. Epub 2010 September 01.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment ( mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein - chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies.

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Background. Some skin carcinomas may be very aggressive. Intensity of angiogenesis, measured by intratumoral vessel density using expression of CD34, has been associated with tumor aggressiveness. In this study, the expression of CD34 in basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) with skull base invasion was compared with that in tumors with good outcome.Methods. Expression of CD34 was graded as mild, moderate, and intense, in 24 BCCs and 11 SCCs with skull base invasion. The control group included 23 BCCs and 10 SCCs.Results. Intense expression of CD34 was noted in 25.00% of BCCs with skull base invasion, compared with 4.35% in the control group (p =.058). Regarding SCCs, intense expression of CD34 was found in 54.55% of aggressive tumors, compared with 10.00% in the control group (p = 133).Conclusions. A trend toward denser microvascular angiogenesis was observed in both BCCs and SCCs with skull base invasion compared with less aggressive controls. (C) 2004 Wiley Periodicals, Inc.