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Biomedical subjects

F Gonzales

Publications and source records attributed to F Gonzales.

22 records · Page 2Linked to original sources

Invasion of an artificial blood vessel wall by human fibrosarcoma cells.

Artificial blood vessel walls constructed by the addition of bovine arterial endothelial cells to multilayers of rat smooth muscle cells were used as substrates for the human fibrosarcoma cell line HT1080. The extracellular matrix proteins elaborated by the smooth muscle cells were prelabeled with [3H]-proline; therefore, their subsequent digestion could be followed by the appearance of radioactivity in the culture medium. The fibrosarcoma cells rapidly hydrolyzed smooth muscle multilayers in the absence of endothelial cells, but an endothelial layer markedly retarded the destructive ability of the tumor cells. The protective effect of the endothelium was not due to a lack of penetration of this cell layer, since HT1080 cells were observed by light and electron microscopy to be in the subendothelial area 24 hr after plating. Subsequently, the tumor cells multiplied in the region between the endothelial and smooth muscle layers and, although their degradative ability was retarded, they were ultimately capable of destroying the structure. Endothelial cells also inhibited hydrolysis of the smooth muscle layers if added simultaneously or up to 1 week after HT1080 cells, but the degree of inhibition was not as great as that seen with a preestablished endothelial layer. Measurable inhibition of tumor cell degradative activity was observed at fibrosarcoma:endothelial cell ratios of 25:1, demonstrating the potency of endothelial cells in modulating this aspect of the invasive phenotype. Although the HT1080 cells only slowly degraded the preexisting matrix proteins in artificial vessel wall cultures, they interfered with the production of new connective tissue proteins which occurred in control cultures. These experiments therefore suggest that endothelial cells have profound effects on tumor cell proteolytic activity, and the significance of these observations to tumor cell extravasation in vivo is discussed.

Animals↗

The relationship of ribosomal RNA synthesis to the formation of segregated nucleoli and nucleolus-like bodies.

The relationship of ribosomal RNA (rRNA) synthesis to nucleolar ultrastructure was studied in partial nucleolar mutants of Xenopus laevis. These mutations are the result of a partial deletion of rRNA genes and therefore alow studies on nucleolar structure and function without using drugs that inhibit rRNA synthesis. Ultrastructural studies demonstrated that normal embryos have reticulated nucleoli that are composed of a loose meshwork of granules and fibrils and a typical nucleolonema. In contrast, partial nucleolar mutants in which rRNA synthesis is reduced to less than 50% of the normal rate have compact nucleoli and nucleolus-like bodies. The compace nucleoli contain granules and fibrils, but they are segregated into distinct regions, and a nucleolonema is never seen. Since other species of RNA are synthesized normally by partial nucleolar mutants, these results demonstrate that nucleolar segragation is related specifically to a reduction in rRNA synthesis. The nucleolus-like bodies are composed mainly of fibrils,and the number of such bodies are composed mainly of fibrils, and the number of such bodies present in the different nucleolar mutants is inversely related to the relative rate of rRNA synthesis. Although the partial nucleolar organizers produce segregated nucleoli in these mutants, they organize morphologically normal, but smaller, nucleoli in heterozygous embryos. Alternative explanations to account for these results are discussed.

Animals↗

Enumeration of waterborne Escherichia coli with petrifilm plates: comparison to standard methods.

Escherichia coli is often monitored in environmental waters as an indicator of the possible presence of human pathogens associated with feces. Petrifilm E. coli/coliform count plates (3M, Minneapolis, MN), previously validated for enumerating E. coli in food, were tested for monitoring E. coli in environmental water. Escherichia coli counts in environmental water samples enumerated with Petrifilm were significantly correlated (R > 0.9; slope = 0.9-1.0; p < 0.001) with counts obtained with three commonly used methods, mTEC (Becton Dickinson, Sparks, MD), m-ColiBlue (Hach, Loveland, CO), and Colilert-18/IDEXX Quanti-Tray 2000 (IDEXX, Westbrook, ME). Blue colonies on Petrifilm plates were most reliably identified as E. coli when accompanied by gas formation, as determined by characterization of the colonies on MacConkey agar plates (PML Microbiologicals, Mississauga, ON, Canada) and by polymerase chair reaction (PCR) with E. coli-specific primers. The main disadvantage of Petrifilm plates for environmental water testing is the small volume (1 mL per sample) that can be tested; however, the plates appear to be suitable for screening and locating sites that exceed criteria for total body and partial body contact. Simplicity of use and storage, reliability, and relatively low cost make Petrifilm plates suitable for volunteer-based and educational water quality monitoring applications, particularly when used as a preliminary screening method to identify problem sites.

DNA, Bacterial↗