RESUMEN
Riparian thickets of East Africa harbor a large number of endemic animal and plant species, but also provide important ecosystem services for the human being settling along streams. This creates a conflicting situation between nature conservation and land-use activities. Today, most of this former pristine vegetation is highly degraded and became replaced by the invasive exotic Lantana camara shrub species. In this study, we analyze the movement behavior and habitat use of a diverse range of riparian bird species and model the habitat availability of each of these species. We selected the following four riparian bird species: Bare-eyed Thrush Turdus tephronotus, Rufous Chatterer Turdoides rubiginosus, Zanzibar Sombre Greenbul Andropadus importunus insularis, and the Kenyan endemic Hinde's Babbler Turdoides hindei. We collected telemetric data of 14 individuals during a 2 months radio-tracking campaign along the Nzeeu River in southeast Kenya. We found that (1) all four species had similar home-range sizes, all geographically restricted and nearby the river; (2) all species mainly use dense thicket, in particular the invasive L. camara; (3) human settlements were avoided by the bird individuals observed; (4) the birds' movement, indicating foraging behavior, was comparatively slow within thickets, but significantly faster over open, agricultural areas; and (5) habitat suitability models underline the relevance of L. camara as suitable surrogate habitat for all understoreyed bird species, but also show that the clearance of thickets has led to a vanishing of large and interconnected thickets and thus might have negative effects on the population viability in the long run.
RESUMEN
BACKGROUND: Gene signatures of sporadic colorectal carcinoma tissues and microdissected colorectal tumor cells were analyzed to identify stromal and tumor cell-specific markers, respectively. METHODS: Serial sections of frozen colorectal tumors (n=29) were subjected to RNA isolation of (1) entire tissue sections with a various tumor cell content and of (2) microdissected invasive tumor cells. Three matching samples of microdissected normal colorectal epithelial and invasive tumor cells were similarly obtained. RNA samples were analyzed using the HG95A and HG95Av2 GeneChip microarrays (Affymetrix). The microarray data was evaluated by established methods and validated by Q-RT-PCR. RESULTS: Unsupervised hierarchical cluster analysis of 18 sample pairs (training set) clearly distinguished tumors from microdissected tumor cells. A 149-gene signature was identified using statistical methods, which was then validated by a hierarchical clustering analysis of 11 independent sample pairs (test set). Genes specifically associated with microdissected invasive tumor cells were for example CKS2 and NME1. In contrast, genes associated with stromal cells were for example MMP2, SDF1 and FBLN2. Finally, a 65-gene signature distinguished normal colorectal epithelial cells and invasive tumor cells, including down-regulation of BMP2 and ANPEP mRNA expression as well as up-regulation of TKT, SPARC, MCM5 mRNA expression. CONCLUSIONS: Our approach allowed precise evaluation of molecular signatures in morphologically defined cell populations and identified novel target genes related to stroma-tumor interactions in colorectal cancer. The approach enables further analysis of gene signatures in different tumor areas and cell types, such as within invasive margins to decipher molecular mechanisms of colorectal cancer invasion and metastasis.
Asunto(s)
Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Perfilación de la Expresión Génica/métodos , ARN Neoplásico/análisis , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/genética , Femenino , Marcadores Genéticos , Humanos , Masculino , Microdisección , Persona de Mediana Edad , Invasividad Neoplásica , Análisis de Secuencia por Matrices de Oligonucleótidos , Análisis de Matrices TisularesRESUMEN
Microarray technology has become a standard tool for generation of gene expression profiles to explore human disease processes. Being able to start from minute amounts of RNA extends the fields of application to core needle biopsies, laser capture microdissected cells, and flow-sorted cells. Several RNA amplification methods have been developed, but no extensive comparability and concordance studies of gene expression profiles are available. Different amplification methods may produce differences in gene expression patterns. Therefore, we compared profiles processed by a standard microarray protocol with three different types of RNA amplification: (i) two rounds of linear target amplification, (ii) random amplification, and (iii) amplification based on a template switching mechanism. The latter two methods accomplish target amplification in a nonlinear way using PCR technology. Starting from as little as 50 ng of total RNA, the yield of labeled cRNA was sufficient for hybridization to Affymetrix HG-U133A GeneChip array using the respective methods. Replicate experiments were highly reproducible for each method. In comparison with the standard protocol, all three approaches are less sensitive and introduced a minor but clearly detectable bias of the detection call. In conclusion, the three amplification protocols used are applicable for GeneChip analysis of small tissue samples.