CA IX is a tumour-associated carbonic anhydrase with proposed roles in pH modulation and intercellular communication. Its distribution was examined in normal, benign and malignant breast tissues and compared with expression of breast tumour markers including oestrogen receptor, c-erbB2, c-erbB3 and CD44. Tissue specimens were analysed using immunohistochemistry and/or reverse transcriptase-polymerase chain reaction (RT-PCR). CA IX was detected by IHC in 12/26 (46%) malignant tissues, 4/36 (11%) benign lesions, but not in 10 normal breasts. Staining was mostly confined to plasma membranes of abnormal epithelial cells, but in five cases was found in adjacent stroma. Semi-quantitative RT-PCR detected CA9 mRNA in 25/39 (64%) malignant tumours, 11/33 (33%) benign lesions, but in none of three normal breasts. Comparative RT-PCR analysis of malignant tissues revealed a relationship between CA9 positivity and c-erbB2 overexpression (p=0.05). Moreover, CA9-positive specimens displayed a significantly higher median level of c-erbB2 than CA9-negative ones (p=0.02). No significant association was found with the other markers. The results of this study support the possible importance of CA IX for breast carcinogenesis and suggest its potential use as a breast tumour marker.
Amifostine protects normal cells from DNA damage induction by ionizing radiation or chemotherapeutics, whereas cancer cells typically remain uninfluenced. While confirming this phenomenon, we have revealed by comet assay and currently the most sensitive method of DNA double strand break (DSB) quantification (based on γH2AX/53BP1 high-resolution immunofluorescence microscopy) that amifostine treatment supports DSB repair in γ-irradiated normal NHDF fibroblasts but alters it in MCF7 carcinoma cells. These effects follow from the significantly lower activity of alkaline phosphatase measured in MCF7 cells and their supernatants as compared with NHDF fibroblasts. Liquid chromatography-mass spectrometry confirmed that the amifostine conversion to WR-1065 was significantly more intensive in normal NHDF cells than in tumor MCF cells. In conclusion, due to common differences between normal and cancer cells in their abilities to convert amifostine to its active metabolite WR-1065, amifostine may not only protect in multiple ways normal cells from radiation-induced DNA damage but also make cancer cells suffer from DSB repair alteration.
) and organic nitrogen (DON) were measured in cloud water samples collected over the northern lower peninsula of Michigan. Within a given cloud field, several altitudes were sampled to examine changes in concentration and speciation with altitude. Several samples were analyzed for bacterial content and activity. Convective cumulus (cumulus congestus) were more concentrated than fair weather cumulus (cumulus humilis) for all major ions and DON, with the cloudy air DON concentrations in convective cumulus being twice as large as for fair weather cumulus, and for all other ions, the droplets were 4-6 times more concentrated. The molar average distribution of nitrogen in the cloud water was 43 (±10, 1s)% ammonium, 39 (±7)% nitrate and 18 (±11)% DON. High concentrations of bacteria were observed in the clouds with an average concentration of 2.9 Â 10 5 (±1.0 Â 10 5 , 1s) bacteria m À3 of cloudy air but which contributed less than 1% of the nitrogen in the cloud water. In addition, nitrifying bacteria were identified, indicating bacterial processing of nitrogen in the cloud water may occur. Air mass origin and altitude influence observed cloud water concentrations, with the exception of DON. The correlation of ammonium and sulfate, and calcium and nitrate suggest that ammonium sulfate and calcium nitrate aerosol may be important sources of these ions.
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