Selecting antibodies to detect HER2 overexpression by immunohistochemistry in invasive mammary carcinomas.

There is an increasing clinical demand for HER2 analysis in breast cancer, especially since the release of trastuzumab. The authors assessed the ability of immunohistochemistry to detect HER2 overexpression in invasive mammary carcinomas (IMC) using five antibodies. Paraffin-embedded samples of 86 IMCs (T2N0) were used to compare the immunohistochemical overexpression of HER2 using two polyclonal antibodies (HercepTest [DAKO] and A0485 [DAKO]) and three monoclonal antibodies (CB11 from two different laboratories, Biogenex and Novocastra, and 4D5 [Genentech]). All immunostainings were scored according to the FDA-approved HercepTest recommendations. The HercepTest-positive cases were compared with gene amplification by FISH (Oncor Inform, Ventana). The HercepTest was positive in 31 of the 86 cases (36.1%). The DAKO antibody A0485 was positive in 25 of the 66 (37.8%). Monoclonal antibody 4D5 was positive in only 15 of the 86 cases (17.4%). There was almost total agreement in results between the two CB11 antibodies: 25 of the 86 positive cases (29.1%). All cases positive for CB11 or 4D5 were HercepTest positive. Most of the HercepTest 2+ cases were negative when using either monoclonal antibody. FISH was positive in 19 of the 20 HercepTest 3+ cases and negative in 5 HercepTest 2+ cases. Three CB11-2+ cases showed no amplification by FISH. In three FISH-positive cases the immunohistochemistry showed no overexpression by all antibodies used. These findings suggest that immunohistochemistry may be used reliably as a primary methodology for evaluating HER2; however, the use of polyclonal antibodies may not be adequate to assess HER2 overexpression. CB11, regardless of the manufacturer (Biogenex or Novocastra), showed better concordance with FISH (kappa=0.83) than did the polyclonal antibodies.     

Facilitating written work using computer word processing and word prediction.

OBJECTIVE: The purpose of this study was to investigate whether occupational therapy intervention that focused on teaching children to use word processing, either alone or with word prediction, was effective in improving the written communication skills of children with learning disabilities and handwriting problems. METHOD: A single-subject alternating treatments design was replicated across three children in grades 4 and 5. During the baseline phase the children wrote stories by hand; during the intervention phase, the children wrote stories, alternating among handwriting, word processing, and word processing with word prediction. Dependent variables focused on percentages of legible words, percentages of correctly spelled words, total amount written, and rate of writing. Data were analyzed by visual inspection. RESULTS: Results were variable. Two children had clear improvements in legibility when using either word processing alone or with word prediction. These same children demonstrated clear improvements in spelling when using word prediction. Though rate of writing was best for two children when using handwriting, relative to total amount produced, one method was not clearly preferable to another. CONCLUSION: Occupational therapy intervention involving word processing with word prediction improves the legibility and spelling of written assignments completed by some children with learning disabilities and handwriting difficulties. It is important to evaluate each child individually and provide training and ongoing support for technology use.     

Can a leukocyte depletion filter (LDF) reduce the risk of reintroduction of hepatocellular carcinoma cells?

During liver transplantation for hepatocellular carcinoma (HCC) patients, HCC could theoretically be introduced into the systemic circulation when salvaged blood is used with an autotransfusion device. Several reports have shown that some types of leukocyte depletion filters (LDFs) could completely reduce the risk for reintroducing some types of tumor cells. In this study, we tested the ability of the LDF (RCEZ1T, Pall Biomedical Co, NY, USA) to reduce the risk for reintroducing HCC cells in vitro by using a very sensitive detection method. We divided the test group into 6 groups: group I was 10 cells, group II was 20 cells, group III was 2 x 10(3) cells, group IV was 2 x 10(5) cells, group V was 2 x 10(6) cells, and group VI was 2 x 10(7) cells. The counted cells in 200 mL saline were passed through the RCEZ1T using the force of gravity. To identify the presence of cells, the pellet was resuspended, and polymerase chain reaction (PCR) was performed. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a housekeeping gene, was used as a primer. In groups I and II, the HCC cells were completely filtered in all experiments. However, in groups III, IV, and V, the HCC cells were not completely filtered in a few of the repeated experiments, with the unfiltered rate of tumor cells being between 8% and 20%. In group VI, the HCC cells were not completely filtered in all the repeated experiments. In conclusion, the RCEZ1T filter markedly reduced the risk for reintroduction of HCC cells. However, at high HCC cell load the filter cannot completely remove all the tumor cells. Further studies are required to assess the impact in clinical settings.     

Expression of Epidermal Growth Factor Receptors in Human Breast Cancer: Mass Versus Ligand Binding Capacity

Abstract: Thirty percent of lymph node-negative women with primary breast cancers are at high risk for early recurrence of metastatic disease and diminished survival. Identification of these high-risk patients is a long-term objective of our laboatory, and the epidermal growth factor receptor (EGFR) was investigated as a prognostic factor, EGFR was measured by a ligand binding assay developed in house, in the competition mode that served as the "gold standard" for assessing receptor content and activity. In contrast, measurements of mass (content) were performed by an enzyme immunoassay (EIA) from Ciba Corning Diagnostics (Alameda, CA) and by an enzyme-linked immunosorbent assay (ELISA) from Oncogene Science (Uniondale, NY). A total of 78 breast carcinomas were examined. The median binding capacity measured with [¹²⁵I]EGF was 13 fmol/mg membrane protein (range 0–981), that measured by EIA was 8 fmol/mg (range 1–125), while EGFR measured by ELISA was 135 (range 0–751). Distribution of EGFR did not appear to vary as a function of patient age. Neither the results from EIA nor those from ELISA correlated with those obtained by the ligand binding assay. However, there was a good correlation of results obtained by the two antibody-based assays despite the fact that the calibration of standards was considerably different. These data suggest that EGFR should be measured by ligand binding assay for clinical studies; mass assays based on antibody reagents will require further development.     

Postnephrectomy arteriovenous fistula of the renal pedicle treated with detachable balloons: A case report

A case of postnephrectomy arteriovenous fistula of the right renal pedicle is reported here. The diagnosis was confirmed by angiography, and successful treatment was achieved using detachable balloon.     

Natural history of atopic dermatitis and its relationship to serum total immunoglobulin E in a population-based birth cohort study

We investigated the natural history of atopic dermatitis (AD) in a population-based birth cohort and assessed whether children at risk of visible eczema at 5 years of age can be identified from total immunoglobulin E (IgE) levels measured at 8, 12 and 18 months. AD data collected included a whole body examination for visible eczema at 49 months (4 years) and 61 months (5 years) of age and parent completed questionnaire data throughout their early lives. Children were divided into four groups based on their natural history of early AD: persistent (AD at 1, 6, 18, 30 and 42 months, n  = 34), intermittent early onset (before 18  months of age, n  = 495), intermittent late onset (18–42 months of age, n  = 273) and unaffected ( n  = 429). Visible eczema at 5 years of age was present in 12.2% (117/957) (95% confidence interval [CI] 10.1–14.3%) of the children. Levels of total IgE at 8, 12 and 18 months of age were associated with early onset of AD, but not with AD of later onset. For all four natural history groups, the geometric mean total IgE at 12 months was higher in those who subsequently had visible eczema than those who did not. However, the degree of overlap was such that total IgE at 12 months of age was a poor predictor of eczema at age five. A cutoff point of 78 kU/l had the highest positive predictive value for visible eczema at 5 years of age of 28.6%, with a sensitivity of 12% and specificity of 95%.     

Periosteal remodeling at the femoral neck in nonhuman primates.

Periosteal bone turnover is poorly understood. We documented intramembranous periosteal bone turnover in the femoral neck in intact nonhuman primates and an increase in osteoclast numbers at the periosteal surface in sex steroid-deficient animals. Our studies are the first to systematically document periosteal turnover at the femoral neck. INTRODUCTION: Bone size is an important determinant of bone strength, and cellular events at the periosteal surface could alter bone dimensions. We characterized periosteal cellular activity with dynamic histomorphometric studies of nonhuman primate femoral neck and shaft. MATERIALS AND METHODS: Femur specimens from 16 intact adult male and female nonhuman primates (Rhesus [Macaca mulatta, n = 9] and Japanese Macaque [Macaca fuscata, n = 7]) were analyzed. Animals were double-labeled with tetracycline, and necropsy was performed 2-7 days after the last dose. We characterized periosteal resorptive activity in an additional group of five intact and four castrate female animals. Multiple group comparisons in intact animals were performed by one-way ANOVA followed by a Fisher PLSD posthoc test. In gonadectomized animals, Fisher's exact test was used for dichotomous and Mann-Whitney U-test for continuous variables. RESULTS: Bone turnover in the periosteum of the femoral neck in intact animals was more rapid than at the femoral shaft but slower than in femoral neck cancellous bone. Similarly, in these intact animals, the eroded surface of cortical bone at the femoral neck periosteal surface was significantly greater than in the cancellous bone compartment (p < 0.0001) or on the femoral shaft (p < 0.0001). Gonadectomized female animals showed an increase in osteoclast number on the periosteal surface compared with intact controls (p < 0.01). CONCLUSIONS: We documented intramembranous periosteal bone turnover in the femoral neck by histomorphometric analyses. The tissue level bone formation rate was sufficient to add substantively to femoral neck size over time. Periosteal osteoclastic activity was not the result of the emergence of intracortical tunneling at the bone surface. Sex steroid deficiency produced an increase in osteoclast numbers at the periosteal surface. This is the first systematic documentation of periosteal turnover at the femoral neck.