Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

<p>Abstract</p> <p>Background</p> <p><it>HER-2 </it>gene testing has become an integral part of breast cancer patient diagnosis. The most commonly used assay in the clinical setting for evaluating HER-2 status is immunohistochemistry (IHC) and fluorescence <it>in situ </it>hybridization (FISH). These procedures permit correlation between <it>HER-2 </it>expression and morphological features. However, FISH signals are labile and fade over time, making post-revision of the tumor difficult. CISH (chromogenic <it>in situ </it>hybridization) is an alternative procedure, with certain advantages, although still limited as a diagnostic tool in breast carcinomas.</p> <p>Methods</p> <p>To elucidate the molecular profile of HER-2 status, mRNA and protein expression in 75 invasive breast carcinomas were analyzed by real time quantitative RT-PCR (qRT-PCR) and IHC, respectively. Amplifications were evaluated in 43 of these cases by CISH and in 11 by FISH.</p> <p>Results</p> <p>The concordance rate between IHC and qRT-PCR results was 78.9%, and 94.6% for qRT-PCR and CISH. Intratumoral heterogeneity of <it>HER-2 </it>status was identified in three cases by CISH. The results of the three procedures were compared and showed a concordance rate of 83.8%; higher discordances were observed in 0 or 1+ immunostaining cases, which showed high-level amplification (15.4%) and <it>HER-2 </it>transcript overexpression (20%). Moreover, 2+ immunostaining cases presented nonamplified status (50%) by CISH and <it>HER-2 </it>downexpression (38.5%) by qRT-PCR. In general, concordance occurred between qRT-PCR and CISH results. A high concordance was observed between CISH/qRT-PCR and FISH. Comparisons with clinicopathological data revealed a significant association between <it>HER-2 </it>downexpression and the involvement of less than four lymph nodes (<it>P </it>= 0.0350).</p> <p>Conclusion</p> <p>Based on these findings, qRT-PCR was more precise and reproducible than IHC. Furthermore, CISH was revealed as an alternative and useful procedure for investigating amplifications involving the <it>HER-2 </it>gene.</p

The Effects of Land Use on the Mobility of Elderly and Disabled and Their Homecare Workers, and the Effects of Care on Client Mobility: Findings from Contra Costa, California

This study looks at the relationships among land use; the mobility of disabled and elderly recipients of public home healthcare; the mobility of their homecare workers; and how much care those homecare workers provide. The findings are based on nearly 1,300 survey responses from clients and homecare workers in the In-Home Supportive Services (IHSS) program in Contra Costa County, California, a publicly funded program for individuals with disabilities who have low incomes. The homecare workers I surveyed belong to the Service Employees International Union (SEIU). The qualitative data and descriptive statistics paint a portrait of both populations’ transportation habits and challenges. Regression analyses, controlling for variables such as car ownership, disability level, gender, age, and race, tested the interactions between the variables of interest in six hypotheses.The results are complex and occasionally conflicting, yet patterns appear. For example, the IHSS clients have car-use rates far lower than average, with only 10% driving themselves when they leave home, and almost half live alone; these facts, combined with their low incomes and disabilities, mean that IHSS clients are sensitive to how much transportation assistance they receive in terms of how often they leave home and what destinations they are able to reach. They also respond to land use characteristics, especially when measured at the neighborhood scale, with those living in higher density and accessibility areas generally experiencing greater mobility. The homecare workers similarly have low incomes and use alternative modes of transportation more often than do Contra Costa commuters on average. Unlike their clients, homecare workers living in higher density and accessibility areas generally experienced increased travel challenges. But living closer to their clients was associated with being able to provide more effective care, as was having an easier commute measured by other variables. The more care provided, the greater mobility their clients experienced.The populations of care recipients and professional homecare workers are growing as, among other trends, the proportion of senior citizens increases and families disperse across the country or world. Understanding mobility barriers as well as ways to facilitate efficient and effective care provision becomes all the more important. This study describes transportation problems that IHSS clients and caregivers encounter and points to certain possible responses, in particular expanding the transportation assistance that caregivers are able to provide