RESUMO
BACKGROUND: The molecular messages which drive angiogenesis in the adventitia of an aneurysmal aorta are uncertain. The emergence of molecular phenotyping by cDNA expression arrays provides a simple and rapid method for a preliminary approach to the analysis of molecular messengers for neovascularization in cultured cells. In the present experiment, fibroblasts cultured from the aorta of a patient with abdominal aortic aneurysm (AAA) were compared with normal dermal fibroblasts, on an array that evaluates several mRNAs with known roles in angiogenesis. METHODS: RNA was isolated from fibroblasts and purified. Labelled cDNA probes were generated from a mixture of RNA and CDS primers. Atlas Array membranes (Clontech) were prehybridized by ExpressHyb buffer. The cDNA probes were then added to the membranes, which were exposed to Phospholmage Screen (Molecular Dynamics) and analysed by a dedicated computer program. RESULTS: The most significantly upregulated mRNAs in AAA (by comparison to dermal fibroblasts) were: MCAF, MDNCF, EGR-1, VEGF, FGF-7, Mal protein, Mac Marcks, Transducin, Interleukin-9 receptor, and TNF. CONCLUSION: VEGF and TNF were upregulated, as expected. However, the upregulation of monocyte chemotactic and activating factor (MCAF) and monocyte-derived neutrophil chemotactic factor (MDNCF) suggest that the fibroblast may be more significantly involved in driving the inflammatory response that leads to AAA than previously realized.
Assuntos
Aneurisma da Aorta Abdominal/genética , Aneurisma da Aorta Abdominal/fisiopatologia , Fibroblastos/fisiologia , Expressão Gênica , Neovascularização Fisiológica/genética , Fenômenos Fisiológicos da Pele , Aneurisma da Aorta Abdominal/patologia , Células Cultivadas , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/metabolismo , Pele/citologiaRESUMO
The response characteristics of some iodide-selective solvent polymeric membrane electrodes based on with N,N'-bis(salicylaldehyde-n-octyl) diimine cobalt(II) (Co(II)SAODI) which is a more lipophilic substitute for a previously reported iodide-carrier are described. The electrode doped with Co(II)SAODI into a plasticized poly(vinyl chloride) membrane exhibits an anti-Hofmeister selectivity pattern with high selectivity toward iodide, long lifetime and small interference from H(+). Quartz crystal microgravimetric measurements and ac impedance experiments show that the excellent selectivity for iodide is related to the unique interaction between the carriers and iodide and steric effect associated with the structure of the Schiff base ligands.
RESUMO
Halo assay, a method for quantitative analysis of chromatin structure in cell, was developed recently. Due to visualization, feasibility, rapidity, and sensitivity of the method, it has been applied in radiobiology and cancer research. According to previous reports, with some improvement, this method has been established in our laboratory. Employing halo assay, the chromatin structure of two kinds of cell strain with similar genetic background and different radiosensitivity was analyzed and the results show that radiosensitive strain has looser chromatin structure than radioresistant one. It is possible that this difference is related to their different radiosensitivity, thus it's a proof for the relation of chromatin structure and radiosensitivity.
Assuntos
Cromatina/química , Animais , Processamento de Imagem Assistida por Computador , Microscopia de Fluorescência , Neoplasias Experimentais/patologia , Tolerância a Radiação , Células Tumorais Cultivadas/químicaRESUMO
Fluorometric method was modified and used to detect DNA interstrand crosslinks (ISC) in tumor cells after treatment with cisplatin. Linear dose-response curve was obtained. The difference of DNA ISC formation between HeLa S3 (containing O6-methylguanine methyltransferase, Mer+) and HeLa MR (devoid of O6-methyguanine methyltransferase, Mer-) cells was studied after treatment with alkylating agent 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride (ACNU). The survival fraction was also observed in Mer+ and Mer- cells treated with ACNU. It seems that DNA ISC formation may be used as one of the possible criteria in estimating the sensitivity of tumor cells to bifunctional alkylating agent and forecasting the efficacy of tumor chemotherapy.