A comprehensive differential gene expression screen on a panel of 54 breast tumors and >200 normal tissue samples using DNA microarrays revealed 15 genes specifically overexpressed in breast cancer. One of the most prevalent genes found was trichorhinophalangeal syndrome type 1 (TRPS-1), a gene previously shown to be associated with three rare autosomal dominant genetic disorders known as the trichorhinophalangeal syndromes. A number of corroborating methodologies, including in situ hybridization, e-Northern analysis using ORF EST (ORESTES) and Unigene EST abundance analysis, immunoblot and immunofluorescence analysis of breast tumor cell lines, and immunohistochemistry, confirmed the microarray findings. Immunohistochemistry analysis found TRPS-1 protein expressed in >90% of early-and late-stage breast cancer, including ductal carcinoma in situ and invasive ductal, lobular, and papillary carcinomas. The TRPS-1 gene is also immunogenic with processed and presented peptides activating T cells found after vaccination of HLA-A2.1 transgenic mouse. Human T cell lines from HLA-A*0201 ؉ female donors exhibiting TRPS-1-specific cytotoxic T lymphocyte activity could also be generated.gene expression profiling ͉ immunohistochemistry

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Assessment of human epidermal growth factor receptor-2 status is standard practice in women with breast cancer. Most laboratories use immunohistochemistry as a screening test, with equivocal results confirmed by fluorescence in-situ hybridization (FISH). Chromogenic in-situ hybridization (CISH) is a relatively new method for detection of gene amplification using a peroxidase reaction, which can be viewed using a standard light microscope. This study was undertaken to validate CISH as a method for assessing human epidermal growth factor receptor-2 gene amplification. The gene amplification status of human epidermal growth factor receptor-2 immunohistochemistry negative (0/1 þ , n ¼ 69; Group 1), immunohistochemistry positive (3 þ , n ¼ 50; Group 2) and equivocal tumor samples (2 þ , n ¼ 135; Group 3) was evaluated by FISH and CISH, and the concordance between FISH and CISH results calculated. In Group 1, 67/69 cases did not show amplification by CISH and 69/ 69 showed no amplification by FISH. Two cases were discordant; therefore, fluorescence/CISH concordance was 97%. In Group 2, 46/50 cases were amplified by FISH and 47/50 cases were amplified by CISH; three cases were not amplified by either method (immunohistochemistry false-positives). Only one case showed discordant FISH and CISH results, making the fluorescence/CISH concordance 98%. In Group 3, 89/135 cases were not amplified and 37/135 were amplified by both methods. Nine cases were discordant, giving a fluorescence/CISH concordance of 93%. The discordant cases were those with very low or borderline amplification with FISH. The high level of concordance between FISH and CISH seen in this study suggests that CISH may be a viable alternative to FISH for use in the human epidermal growth factor receptor-2 testing algorithm.

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Toxicity data for tropical species are often lacking for ecological risk assessment. Consequently, tropical and subtropical countries use water quality criteria (WQC) derived from temperate species (e.g., United States, Canada, or Europe) to assess ecological risks in their aquatic systems, leaving an unknown margin of uncertainty. To address this issue, we use species sensitivity distributions of freshwater animal species to determine whether temperate datasets are adequately protective of tropical species assemblages for 18 chemical substances. The results indicate that the relative sensitivities of tropical and temperate species are noticeably different for some of these chemicals. For most metals, temperate species tend to be more sensitive than their tropical counterparts. However, for un-ionized ammonia, phenol, and some pesticides (e.g., chlorpyrifos), tropical species are probably more sensitive. On the basis of the results from objective comparisons of the ratio between temperate and tropical hazardous concentration values for 10% of species, or the 90% protection level, we recommend that an extrapolation factor of 10 should be applied when such surrogate temperate WQCs are used for tropical or subtropical regions and a priori knowledge on the sensitivity of tropical species is very limited or not available.

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Aim: Double-walled carbon nanotubes (DWNTs) are found in a variety of consumer products, but there are no ecotoxicity data of DWNTs into marine organisms. Materials & methods: Chronic toxicity of DWNTs was investigated with the diatom Thalassiosira pseudonana, copepod Tigriopus japonicus and medaka Oryzias melastigma. DWNTs were dispersed using sonication (so-DWNTs) and stirring (st-DWNTs) for comparison. Results: The median aggregation size (0.89 µm2) of so-DWNTs was smaller than that of st-DWNTs (21.8 µm2). Exposure to DWNTs led to growth inhibition of T. pseudonana with EC50s of 1.86 and 22.7 mg/l for so- and st-DWNTs, respectively. Population growth of T. japonicus was reduced to 0.1 mg/l for so-DWNTs and 10 mg/l for st-DWNTs. Growth inhibition in O. melastigma was observed at 10 mg/l for so-DWNTs but not for st-DWNTs. Conclusion: Given that so-DWNTs are consistently significantly more toxic than st-DWNTs, dispersion method and size of aggregations should be considered in DWNT toxicity testing.

Quality standards (QS) for dissolved metals in freshwaters have been proposed underthe European Water Framework Directive (WFD) and are based mainly upon laboratory ecotoxicity data. Uncertainties remain about laboratory-to-field extrapolation to establish QS that are neither over- nor underprotective. Freshwater benthic macroinvertebrates are a group of organisms of known sensitivity to heavy metals. We analyzed a dataset from England and Wales of dissolved metal concentrations (cadmium, chromium, copper, iron, nickel, lead, and zinc) and associated benthic invertebrate community metrics, using piecewise regression, quantile regression, and information on metal concentrations consistent with good quality status. Analysis of these field data suggests that dissolved metal QS proposed under the WFD are similar to metal concentrations in rivers associated with unimpaired benthic invertebrate assemblages in England and Wales. The only exceptions to this are QS for iron and zinc, where use of relatively large assessment factors leads to standards that are substantially below concentrations associated with impaired invertebrate assemblages in the field.