Conventional cytogenetic characterization of a new cell line, ACP01, established from a primary human gastric tumor

Gastric cancer is the second most frequent type of neoplasia and also the second most important cause of death in the world. Virtually all the established cell lines of gastric neoplasia were developed in Asian countries, and western countries have contributed very little to this area. In the present study we describe the establishment of the cell line ACP01 and characterize it cytogenetically by means of in vitro immortalization. Cells were transformed from an intestinal-type gastric adenocarcinoma (T4N2M0) originating from a 48-year-old male patient.This is the first gastric denocarcinoma cell line established in Brazil. The most powerful application of the cell line ACP01 is in the assessment of cytotoxicity. Solid tumor cell lines from different origins have been treated with several conventional and investigational anticancer drugs. The ACP01 cell line is triploid, grows as a single, non-organized layer, similar to fibroblasts, with focus formation,heterogeneous division, and a cell cycle of approximately 40 h. Chromosome 8 trisomy, present in 60% of the cells, was the most frequent cytogenetic alteration. These data lead us to propose a multifactorial triggering of gastric cancer which evolves over multiple stages involving progressive genetic changes and clonal expansion.

Trajetórias tecnológicas e sistemas agrários: qualificação territorial utilizando geoprocessamento em Paragominas e Santarém/PA

A Amazônia tem sido observada principalmente por meio do fenômeno do desmatamento, utilizando recursos tradicionais do sensoriamento remoto como a quantificação de área desflorestada e seu posterior incremento anual, que parece se constituir em uma metodologia eficaz. Ratificando este raciocínio, constatei num levantamento de 16.591 multas aplicadas pelo IBAMA/PA no período de 2000 até 2008, que mais de 85,0% das autuações estavam relacionadas apenas à componente flora;e na jurisdição da Gerência do IBAMA em Santarém, oeste do Pará, em 2008, quase 60% das multas se deu por conta de desflorestamento, identificados via sensoriamento remoto. Ressalta-se que as análises de imagens de satélites por si só não definem elementos da superfície terrestre, contribuindo pouco para o entendimento e posterior intervenção da realidade. Nesse contexto, foram investigados e vetorizados 479 estabelecimentos rurais nas regiões de Paragominas e Santarém, estado do Pará, que possuem históricos de uso e ocupação distintos, qualificando-os segundo suas trajetórias tecnológicas prevalentes, na perspectiva apresentada por Costa, concretizando um importante passo para correção das distorções no desenvolvimento econômico, agregando informação ao dado de sensoriamento remoto. Aplicaram-se recursos geotecnológicos de métricas de paisagem, construíram-se banco de dados celular integrado com estatísticas e algoritmos de otimização probabilística, associando dados de classificação não supervisionada isodata (validadas com kappa= 0,87, classificação considerada“excelente”) com os tipos de produção coletados em campo, gerando uma classificação final "integrada"(kappa= 0,78, classificação “muito boa”). Na região de Paragominas, foram qualificadas 3 tipos de trajetórias tecnológicas, a camponesa T8 (domínio de culturas temporárias), a camponesa T3 e patronal T4 (especializadas em pecuária de corte). Em Santarém, revelaram-se 2 trajetórias, a camponesa T2 (forte presença de culturas permanentes, temporárias e sistemas agro florestais) e a patronal T7 (mutação da T4, aumentando a participação das culturas temporárias). A metodologia aplicada logrou êxito, espacializando as propriedades rurais segundo seus tipos de trajetórias tecnológicas e gerando classes de uso mais representativas, como cultura temporária e pastagem, mas que na classificação via sensoriamento remoto isodata é englobada pela classe “agropecuária”, possibilitando uma visão mais realista das atividades de produção desenvolvidas na área investigada, concretizando a geração de informações espaciais integrando dados de diferentes fontes e o aumento do poder de leitura do pixel.

Propagação de espécies de Annonaceae com estacas caulinares

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência da variação sazonal de atributos de fertilidade do solo sobre a biomassa microbiana em uma cronosseqüencia de plantio de palma de óleo (Elaeis oleifera (Kunth) Cortés) no Nordeste Paraense

O Brasil possui cerca de 32 milhões de hectares de áreas com aptidão para a expansão da cultura da palma de óleo e mais de 90% destas estão na Amazônia. Há necessidade de compreender a interação de novos plantios e de seu desenvolvimento com o meio ambiente. Este estudo foi conduzido em plantios comerciais da empresa Dendê do Pará SA (DENPASA) em Castanhal – Pará e avaliou o efeito da variação sazonal da precipitação sobre a fertilidade do solo e desta sobre a biomassa microbiana do solo em uma cronosseqüencia de plantios de palma de óleo com 5, 8 e 12 anos e floresta secundária. Foram mensurados/estimados e correlacionados os atributos do solo carbono da biomassa microbiana (C_BMS), carbono total (C_TOTAL), nitrogênio (N_TOTAL), respiração basal (RBS), quociente metabólico (qCO₂), relação carbono da biomassa microbiana/ carbono total (C_BMS:C_TOTAL), relação carbono/nitrogênio (C/N), umidade gravimétrica (Ug), fósforo (P), potássio (K), cálcio (Ca), magnésio (Mg), alumínio (Al), capacidade de troca efetiva de cátions (CTC) e saturação por alumínio (m). O C_BMS foi o atributo mais sensível para diferenciar as áreas do estudo e os períodos seco e chuvoso. Os atributos químicos de fertilidade do solo e a biomassa microbiana do solo apresentaram correlações significativas mais fortes e em maior número no período chuvoso. Os índices microbianos qCO₂ e C_BMS:C_TOTAL comprovaram que áreas de plantio convencional podem ser relativamente eficientes em relação à dinâmica do C em comparação a área de floresta secundária.

Protocolo para micropropagação de bananeira ‘Thap maeo’

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Propagação do pinhão manso por estaquia, enxertia e sementes

Pós-graduação em Agronomia - FEIS

Molecular characterization of methicillin-resistant Staphylococcus aureus isolated from a Brazilian university hospital

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

Estatuto do índio, direito indígena e impacto ambiental: aldeias Guarani do Jaraguá (SP)

In Brazil, there are three main codes that surround the issue of right or possession of various lands according to concepts of ownership or purchase. When it comes to indigenous issues in Brazil is difficult to say which legal code applies more fairly the native population of the country. In the case of the Indians who have their reserves near urban areas there is such a conflict of laws becomes more evident and takes even greater than in other regions of Brazil. As is the case in the indigenous villages of the District of Jaragua in Greater São Paulo, the Tekoá YTU and Tekoá Pyau. The two villages are located in northeastern São Paulo and currently are surrounded by continuous growth and disorderly city of Sao Paulo while their inhabitants fight to preserve the customs and traditions of the Guarani people. They prevail on the City Statute (2001), the Indian Statute (EDI) Environmental Laws and the National Council of Environment (CONAMA) and the Forestry Code, the latter solely because they are near the State Park Jaragua - area environmental preservation

Desempenho de tractores florestais processadores

A time-and-motion study in wood processing of the Eucalyptus harvester's operational cycle was developed. The objective was to evaluate wood harvesters working under several site conditions. The research includes two kinds of harvesters while processing wood to supply paper and cellulose factories, as follows: a machine with twin tires and other one with large tracks, both connected to a different head harvester models. The forestry species were Eucalyptus saligna Smith and Eucalyptus grandis Hill ex Maiden, seven and eight years old, in slope terrains ranging from 0% to 10% degrees, under harvesting systems used by Votorantim and Suzano forestry companies, in São Paulo State, Brazi. Considering all field research, results disclosed that, for some conditions, both machines have shown mathematical correlation between some mechanical wood operation within the processing operational cycle and the saw logs production. All the mathematical correlations were found to express logarithmical models.

Mapeamento de solos e da produtividade de plantações de Eucalyptus grandis, com uso de sistema de informação geográfica

Detailed environmental land characterization is essential for technical and financial planning, for both the scientific point of view and technological application. This work aimed at the physiographic and pedological characterization and eucalyptus productivity mapping at Itatinga Forest Sciences Experimental Station (southeastern Brazil), using geographic information systems in order to identify possible cause-effect relationships between forest productivity and soil attributes. The digital cartographic dataset was structured as follows: as primary source of data, aerial photograph and field survey were used and, as a secondary source, topographical, geological and land use occupation maps were used. For mapping wood productivity at age six (MAI6, Mean Annual Increment), inventory data of permanent plots (same species, provenance and age) were used, which were obtained from Eucalyptus grandis plantations. Using simple linear correlation and backward stepwise multiple regression analysis, the dependent variable (MAI) was related with physical and chemical characteristics of the soils. Two standards of contour curves were identified, one with close curves, narrow and surrounding the drainage network, in the steeper and lower altitude areas; the other, with spaced contour lines, in the areas of higher altitude and with plane relief. Six types of soils were characterized as being highly related to the physiographic patterns of the area: loamy sandy to sandy clayey Typic Hapludox (LVAd, 47.5%), clayey Rhodic Hapludox (LVd1, 33.4%), sandy clay Rhodic Hapludox (LVd2, 6%), clayey Rhodic Hapludox (LVdf, 9.1%), Entisols (G, 3.4%) and Fluvents soil (RY, 0.6%). There were large variations in wood productivity in the Eucalyptus grandis plantations, characterized in six classes, ranging from 26 to 52 m(3) ha(-1) yr(-1). These productivity changes were strictly related to soil mapping units. Through multiple regression analysis, we found that clay and organic matter contents were the attributes which most strongly explained the productivity differences.

Novel techniques for the remote sensing of photosynthetic processes

Remote sensing (RS) techniques have evolved into an important instrument to investigate forest function. New methods based on the remote detection of leaf biochemistry and photosynthesis are being developed and applied in pilot studies from airborne and satellite platforms (PRI, solar-induced fluorescence; N and chlorophyll content). Non-destructive monitoring methods, a direct application of RS studies, are also proving increasingly attractive for the determination of stress conditions or nutrient deficiencies not only in research but also in agronomy, horticulture and urban forestry (proximal RS). In this work I will focus on some novel techniques recently developed for the estimation of photochemistry and photosynthetic rates based (i) on the proximal measurement of steady-state chlorophyll fluorescence yield, or (ii) the remote sensing of changes in hyperspectral leaf reflectance, associated to xanthophyll de-epoxydation and energy partitioning, which is closely coupled to leaf photochemistry and photosynthesis. I will also present and describe a mathematical model of leaf steady-state fluorescence and photosynthesis recently developed in our group. Two different species were used in the experiments: Arbutus unedo, a schlerophyllous Mediterranean species, and Populus euroamericana, a broad leaf deciduous tree widely used in plantation forestry. Results show that ambient fluorescence could provide a useful tool for testing photosynthetic processes from a distance. These results confirm also the photosynthetic reflectance index (PRI) as an efficient remote sensing reflectance index estimating short-term changes in photochemical efficiency as well as long-term changes in leaf biochemistry. The study also demonstrated that RS techniques could provide a fast and reliable method to estimate photosynthetic pigment content and total nitrogen, beside assessing the state of photochemical process in our plants’ leaves in the field. This could have important practical applications for the management of plant cultivation systems, for the estimation of the nutrient requirements of our plants for optimal growth.

Recruitment of EB1, a master regulator of microtubule dynamics, to the surface of the Theileria annulata schizont

The apicomplexan parasite Theileria annulata transforms infected host cells, inducing uncontrolled proliferation and clonal expansion of the parasitized cell population. Shortly after sporozoite entry into the target cell, the surrounding host cell membrane is dissolved and an array of host cell microtubules (MTs) surrounds the parasite, which develops into the transforming schizont. The latter does not egress to invade and transform other cells. Instead, it remains tethered to host cell MTs and, during mitosis and cytokinesis, engages the cell's astral and central spindle MTs to secure its distribution between the two daughter cells. The molecular mechanism by which the schizont recruits and stabilizes host cell MTs is not known. MT minus ends are mostly anchored in the MT organizing center, while the plus ends explore the cellular space, switching constantly between phases of growth and shrinkage (called dynamic instability). Assuming the plus ends of growing MTs provide the first point of contact with the parasite, we focused on the complex protein machinery associated with these structures. We now report how the schizont recruits end-binding protein 1 (EB1), a central component of the MT plus end protein interaction network and key regulator of host cell MT dynamics. Using a range of in vitro experiments, we demonstrate that T. annulata p104, a polymorphic antigen expressed on the schizont surface, functions as a genuine EB1-binding protein and can recruit EB1 in the absence of any other parasite proteins. Binding strictly depends on a consensus SxIP motif located in a highly disordered C-terminal region of p104. We further show that parasite interaction with host cell EB1 is cell cycle regulated. This is the first description of a pathogen-encoded protein to interact with EB1 via a bona-fide SxIP motif. Our findings provide important new insight into the mode of interaction between Theileria and the host cell cytoskeleton.

Analysis of the Soehner-Dmochowski strain of Moloney murine sarcoma virus

The Soehner-Dmochowski strain of murine sarcoma virus (MuSV-SD) was derived from a bone tumor of a New Zealand Black (NZB) rat infected with the Moloney strain of MuSV, which carries the gene encoding the v-mos protein. Serial passage of cell-free tumor extracts both decreased the latent period and resulted in osteosarcomas. Cells from a late passage tumor were established in culture, cell-free extracts frozen, and later inoculated into newborn NZB rats. One of the resulting bone tumors was established in culture and clonal cell lines derived, of which S4 was selected for the present study. The objectives of the study were two-fold: an examination of the genetic organization of MuSV-SD, and an examination of the biochemical characteristics of the viral proteins, since this is an acutely transforming virus which may yield insights into the mechanism of transformation caused by the v-mos protein. Blot hybridization of digested S4 genomic DNA reveals three candidate MuSV-SD integrated viral DNAs. The largest of these, MuSV-SD-6.5, was cloned from an S4 cosmid library, and the complete MuSV-SD-mos sequence was determined. The predicted amino acid sequence of the v-mos protein was compared to that of MuSV-124 and Ht-1, which show a 96.5% and 97.1% similarity, respectively. To characterize the MuSV-SD-mos protein further, immunochemical assays were performed using anti-mos antisera. The immunoblot analysis and immunoprecipitation assays demonstrated that similar levels of the v-mos protein were present in cells chronically infected with either MuSV-SD or MuSV-124; however, the immune complex kinase assay revealed greatly reduced in vitro serine kinase activity of the MuSV-SD-mos protein compared to that of MuSV-124. Sequence analysis demonstrated that the serine at amino acid residue 358 of the MuSV-SD-mos protein, like that of MuSV-Ht-1, had been mutated to a glycine. Mutations of this serine residue have been shown to affect the detectable in vitro kinase activity, however, v-mos proteins containing this mutation still retain transforming properties. Therefore, although the characteristic in vitro kinase activity of the MuSV-SD-mos protein has not been demonstrated, it is clear that this virus is a potent transforming agent. ^

Gradual Phenotypic Conversion Associated with Immortalization of Cultured Human Mammary Epithelial Cells

Examination of the process of immortal transformation in early passages of two human mammary epithelial cell (HMEC) lines suggests the involvement of an epigenetic step. These lines, 184A1 and 184B5, arose after in vitro exposure of finite lifespan 184 HMEC to a chemical carcinogen, and both are clonally derived. Although early-passage mass cultures of 184A1 and 184B5 maintained continuous slow growth, most individual cells lost proliferative ability. Uniform good growth did not occur until 20–30 passages after the lines first appeared. Early-passage cultures expressed little or no telomerase activity and telomeres continued to shorten with increasing passage. Telomerase activity was first detected when the telomeres became critically short, and activity levels gradually increased thereafter. Early-passage cultures had little or no ability to maintain growth in transforming growth factor-β (TGFβ); however, both mass cultures and clonal isolates showed a very gradual increase in the number of cells displaying progressively increased ability to maintain growth in TGFβ. A strong correlation between capacity to maintain growth in the presence of TGFβ and expression of telomerase activity was observed. We have used the term “conversion” to describe this process of gradual acquisition of increased growth capacity in the absence or presence of TGFβ and reactivation of telomerase. We speculate that the development of extremely short telomeres may result in gradual, epigenetic-based changes in gene expression. Understanding the underlying mechanisms of HMEC conversion in vitro may provide new insight into the process of carcinogenic progression in vivo and offer novel modes for therapeutic intervention.

Forest health conditions in Alaska.

Latest issue consulted: 2004.