Characterizing Meat- and Milk/Dairy-like Vegetarian Foods and Their Counterparts Based on Nutrient Profiling and Food Labels.

Vegetarian foods are plant-based (PB) foods, often perceived as healthier foods than animal-based (AB) foods. The objective of this study was to analyze the nutritional quality of a set of PB foods (meat, milk and dairy products) marketed in Spain, and to compare their nutrient profiles with respect to some AB counterparts. Nutritional information per 100 g or mL, ingredients, and nutritional declarations, as well as the Nutri-Score, NOVA, and Eco-Score of each food were collected from Open Food Facts. Differences in the nutrient compositions between PB foods and their counterparts, and between the different groups of PB foods, were assessed at a 5% significance level. A total of 544 PB foods and 373 AB foods were identified. Overall, PB foods had a higher median content of fiber and carbohydrates, but a lower amount of proteins (except PB "meat" analogues: 14 g) and saturated fats (except PB "cheese alternatives": 12.5 g), than the AB counterparts (p < 0.05). PB "milk alternatives", particularly oat "milk", showed a higher median content of total carbohydrates (8 g) and sugars (5.5 g) compared to cow milks (4.7 g carbohydrates/sugars, on average; p < 0.001). PB "meat alternatives" also had a significantly higher value of carbohydrates (9 g) than AB meats (2 g, on average; p < 0.001). PB foods were mostly classified as Nutri-Score A and B (86%). However, more than half of them were of NOVA groups 3 and 4. Thus, there is a great diversity of PB meat and milk/dairy product alternatives on the Spanish market. Despite being products of good nutritional quality compared to AB foods, they also carry drawbacks that could have an impact on nutritional health.

Vitamin D Deficiency and COVID-19: A Biological Database Study on Pathways and Gene-Disease Associations.

Vitamin D (VD) is a fat-soluble vitamin, and pivotal for maintaining health. Several genetic markers have been related to a deficient VD status; these markers could confer an increased risk to develop osteoporosis and other chronic diseases. A VD deficiency could also be a determinant of a severe COVID-19 disease. This study aimed to interrogate genetic/biological databases on the biological implications of a VD deficiency and its association with diseases, to further explore its link with COVID-19. The genetic variants of both a VD deficiency and COVID-19 were identified in the genome-wide association studies (GWAS) catalog and other sources. We conducted enrichment analyses (considering corrected p-values < 0.05 as statistically significant) of the pathways, and gene-disease associations using tools, such as FUMA, REVIGO, DAVID and DisGeNET, and databases, such as the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO). There were 26 and 46 genes associated with a VD deficiency and COVID-19, respectively. However, there were no genes shared between the two. Genes related to a VD deficiency were involved in the metabolism of carbohydrates, retinol, drugs and xenobiotics, and were associated with the metabolic syndrome and related factors (obesity, hypertension and diabetes mellitus), as well as with neoplasms. There were few enriched pathways and disease connections for the COVID-19-related genes, among which some of the aforementioned comorbidities were also present. In conclusion, genetic factors that influence the VD levels in the body are most prominently associated with nutritional and metabolic diseases. A VD deficiency in high-risk populations could be therefore relevant in a severe COVID-19, underlining the need to examine whether a VD supplementation could reduce the severity of this disease.

Characterization and application of Croton blanchetianus Baill extract for lamb ribs preservation.

This research evaluated the antioxidant, antimicrobial and toxicity potential of the leaf extract of Croton blanchetianus Baill (ExCb) and its effect on the conservation of lamb ribs. The ExCb (control treatment) revealed higher concentration of 2,5-dihydroxybenzoic acid (190.10 mg/g), catechin (84.10 mg/g), rosmarinic acid (56.01 mg/g), 4-hydroxybenzoic acid (52.05 mg/g) and myricetin (40.00 mg/g). And it showed high phenolic content (204.05 mg GAE/g), antioxidant potential (11.78 µg/mL by DPPH and 140.40 mmol Sulf Fer/g by FRAP) and antimicrobial activity with inhibition for Staphylococcus aureus, Listeria innocua, Salmonella enterica, Escherichia coli and Aspergillus flavus. However, it showed toxicity against brine shrimp (Artemia Salina) (LD50 of 66.26 µg/mL). The 2,5-dihydroxybenzoic acid was indicated as the main compound responsible for the toxicity of ExCb. After treatment in an oven at 110 °C for 15 min, the toxicity of ExCb was reduced by over 7 times, the compound 2,5-dihydroxybenzoic acid was not identified, and still maintained the phenolic compounds content of 94.35% and antioxidant activity compared to the control (without thermal treatment). The application of absorbent containing 50 mg/mL of ExCb added to the packaging maintained the quality and prevented the lipid oxidation of lamb ribs during 10 days of refrigerated storage.

Rheological and technological characterization of red rice modified starch and jaboticaba peel powder mixtures.

Properties of modified starch and its interaction with functional raw materials are of great interest to the food industry. Thus, this study aimed to evaluate the rheological and technological characterization of starches modified by the action of the enzymes α-amylase and amyloglucosidase and their mixtures with jaboticaba peel powder. The parameters of firmness, gumminess, and final viscosity of starches paste increased, and the tendency to setback was reduced with the addition of jaboticaba peel powder. Starches and mixtures presented shear-thinning behavior. The addition of jaboticaba peel powder to starches increased water, oil, and milk absorption capacity, while syneresis remained stable over the storage period. The addition of jaboticaba peel powder had a positive effect on native and modified starches' rheological and technological properties, qualifying it as an alternative for developing new functional food products.

Influence of enzymatic hydrolysis on the properties of red rice starch.

Starch is a technologically challenging polysaccharide that arouses the global interest in its modification to increase its applicability. Thus, the objective of the present work was to study the influence of enzymatic modification on the properties of red rice starch. The enzymatic modification was performed sequentially through the actions of α-amylase enzymes and amyloglucosidase using a design of experiments where several process conditions were used at sub-gelatinization temperatures. Then the native and modified starches were characterized for its physicochemical, physical, centesimal, phytochemical, morphological, and thermal properties. The enzymatic hydrolysis of the starch granules with α-amylase resulted in 4.46% hydrolysis and more 8.63% after the action amyloglucosidase. Results revealed that surface adsorption is more limiting than diffusion. The enzymatic hydrolysis process resulted in a reduction of the amylase and amylopectin content, as well as on all phytochemicals. After hydrolysis, granules starches presented surface alterations and tendency to become spherical. Small differences on crystallinity revealed that the enzymes had more influence on the amorphous fractions of the starch granules.

Assessing the New American Society of Clinical Oncology/College of American Pathologists Guidelines for HER2 Testing by Fluorescence In Situ Hybridization: Experience of an Academic Consultation Practice.

Context .- Evaluation of HER2 gene amplification by fluorescence in situ hybridization (FISH) was changed by recent American Society of Clinical Oncology/College of American Pathologists (ASCO-CAP) guidelines. Objective . -To determine frequencies and assess patterns of HER2 protein expression for each ASCO-CAP guideline FISH category among 7526 breast cancers accrued to our consultation practice. Design .- We retrospectively reevaluated the HER2 FISH status of breast cancers in our consultation practice according to ASCO-CAP FISH guidelines, and documented HER2 protein levels in each category. Results . -According to new guidelines, 17.7% of our consultation breast cancers were "ISH-positive" with HER2:CEP17 FISH ratios ≥2.0 and average HER2 gene copies per cell ≥4.0 (group 1); 0.4% were "ISH-positive" with ratios ≥2.0 and average copies <4.0 (group 2); 0.6% were "ISH-positive" with ratios <2.0 and average copies ≥6.0 (group 3); 4.6% were "ISH-equivocal" with ratios <2.0 and average copies ≥4.0 and <6.0 (group 4); and 76.7% were "ISH-negative" with ratios <2.0 and average copies <4.0 (group 5). However, only groups 1 (HER2 amplified) and 5 (HER2 not amplified) agreed with our previously reported status, and only these groups demonstrated the expected immunohistochemistry status, overexpression and low expression, respectively. Groups 2 and 4 breast cancers lacked overexpression, whereas group 3 was not significantly associated with either increased or decreased HER2 expression. Conclusions .- Although the status of approximately 95% of our cases (groups 1 and 5) is not affected by the new guidelines, those of the other 5% (groups 2-4) conflict with previous HER2 gene amplification status and with HER2 status by immunohistochemistry.

Hybrid Electrodes by In-Situ Integration of Graphene and Carbon-Nanotubes in Polypyrrole for Supercapacitors.

Supercapacitors also known as electrochemical capacitors, that store energy via either Faradaic or non-Faradaic processes, have recently grown popularity mainly because they complement, and can even replace, conventional energy storage systems in variety of applications. Supercapacitor performance can be improved significantly by developing new nanocomposite electrodes which utilizes both the energy storage processes simultaneously. Here we report, fabrication of the freestanding hybrid electrodes, by incorporating graphene and carbon nanotubes (CNT) in pyrrole monomer via its in-situ polymerization. At the scan rate of 5 mV s(-1), the specific capacitance of the polypyrrole-CNT-graphene (PCG) electrode film was 453 F g(-1) with ultrahigh energy and power density of 62.96 W h kg(-1) and 566.66 W kg(-1) respectively, as shown in the Ragone plot. A nanofibrous membrane was electrospun and effectively used as a separator in the supercapacitor. Four supercapacitors were assembled in series to demonstrate the device performance by lighting a 2.2 V LED.

PTEN Loss Is Associated with Worse Outcome in HER2-Amplified Breast Cancer Patients but Is Not Associated with Trastuzumab Resistance.

PURPOSE: To investigate the clinical relevance of PTEN in HER2-amplified and HER2-nonamplified disease. EXPERIMENTAL DESIGN: We assessed PTEN status in two large adjuvant breast cancer trials (BCIRG-006 and BCIRG-005) using a PTEN immunohistochemical (IHC) assay that was previously validated in a panel of 33 breast cancer cell lines and prostate cancer tissues with known PTEN gene deletion. RESULTS: In the HER2-positive patient population, absence of tumor cell PTEN staining occurred at a rate of 5.4% and was independent of ER/PR status. In contrast, 15.9% of HER2-negative patients exhibited absence of PTEN staining with the highest frequency seen in triple-negative breast cancer (TNBC) subgroup versus ER/PR-positive patients (35.1% vs. 10.9%). Complete absence of PTEN staining in tumor cells was associated with poor clinical outcome in HER2-positive disease. Those patients whose cancers demonstrated absent PTEN staining had a significant decrease in disease-free survival (DFS) and overall survival (OS) compared with patients with tumors exhibiting any PTEN staining patterns (low, moderate, or high). Trastuzumab appeared to provide clinical benefit even for patients lacking PTEN staining. In the HER2-negative population, there were no statistically significant differences in clinical outcome based on PTEN status. CONCLUSIONS: This study is the largest to date examining PTEN status in breast cancer and the data suggest that the rate and significance of PTEN status differ between HER2-positive and HER2-negative disease. Furthermore, the data clearly suggest that HER2-positive patients with PTEN loss still benefit from trastuzumab.

Characterization of HER2 status by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC).

The use of human epidermal growth factor receptor type 2 (HER2) gene amplification and overexpression as a molecular predictive marker has become critically important for proper selection of breast cancer patients for treatment with targeted therapeutic agents such as trastuzumab, lapatinib, pertuzumab, and T-DM1. A high level of sensitivity and specificity of molecular tests for this alteration is desirable. The American Society of Clinical Oncology and College of American Pathology have jointly established consensus guidelines to standardize characterization of this alteration in breast cancers. This chapter provides a brief overview of pre-analytic and analytical processing of breast specimens as well as subsequent molecular evaluation for HER2 status.

Confirmation and further delineation of the 3q26.33-3q27.2 microdeletion syndrome.

Recently, 3 unrelated children with a potentially novel 3q26.33-3q27.2 microdeletion syndrome were reported. We now report a new 9 ½ years old Caucasian boy with a 2 Mb deletion of the same genomic region in combination with Klinefelter syndrome. He presented with facial dysmorphism, developmental delay, Asperger syndrome, thrombocytopenia, recurrent infections and hypogammaglobulinemia. The deletion in our patient improves upon the minimum region of the novel 3q26.33-3q27.2 microdeletion, and provides additional insights into the underlying genetic basis of the observed phenotypes. Consistent with two of three previously described patients, our patient also presents with thrombocytopenia, which we postulate is caused by haploinsufficiency of THPO. In addition, haploinsufficiency of LAMP3, a lymphoid and dendritic cell expressed protein that is implicated in bacterial and viral infections, pulmonary surfactant protein transport and amelogenin degradation, may be a novel cause for the immune deficiency, lung disease and dental abnormalities respectively as seen in these patients.

Alteration of topoisomerase II-alpha gene in human breast cancer: association with responsiveness to anthracycline-based chemotherapy.

PURPOSE: Approximately 35% of HER2-amplified breast cancers have coamplification of the topoisomerase II-alpha (TOP2A) gene encoding an enzyme that is a major target of anthracyclines. This study was designed to evaluate whether TOP2A gene alterations may predict incremental responsiveness to anthracyclines in some breast cancers. METHODS: A total of 4,943 breast cancers were analyzed for alterations in TOP2A and HER2. Primary tumor tissues from patients with metastatic breast cancer treated in a trial of chemotherapy plus/minus trastuzumab were studied for amplification/deletion of TOP2A and HER2 as a test set followed by evaluation of malignancies from two separate, large trials for changes in these same genes as a validation set. Association between these alterations and clinical outcomes was determined. RESULTS: Test set cases containing HER2 amplification treated with doxorubicin and cyclophosphamide (AC) plus trastuzumab, demonstrated longer progression-free survival compared to those treated with AC alone (P = .0002). However, patients treated with AC alone whose tumors contain HER2/TOP2A coamplification experienced a similar improvement in survival (P = .004). Conversely, for patients treated with paclitaxel, HER2/TOP2A coamplification was not associated with improved outcomes. These observations were confirmed in a larger validation set, where HER2/TOP2A coamplification was again associated with longer survival when only anthracycline-containing chemotherapy was used for treatment compared with outcome in HER2-positive cancers lacking TOP2A coamplification. CONCLUSION: In a study involving nearly 5,000 breast malignancies, both test set and validation set demonstrate that TOP2A coamplification, not HER2 amplification, is the clinically useful predictive marker of an incremental response to anthracycline-based chemotherapy. Absence of HER2/TOP2A coamplification may indicate a more restricted efficacy advantage for breast cancers than previously thought.

Acute adrenal crisis in an asthmatic child treated with inhaled fluticasone proprionate.

Adrenal suppression secondary to prolonged inhaled corticosteroid use is usually limited to biochemical abnormalities, with no obvious clinical effects. Acute adrenal crisis is much rarer event but has been reported with increasing frequency. We report a case of a 7-year-old asthmatic child who presented with an acute history of lethargy after a respiratory infection. He was maintained on 220 mug/day of fluticasone propionate for several years. Initial evaluation revealed severe adrenal suppression, with undetectable cortisol levels and minimal response after stimulation with ACTH. After fluticasone was discontinued, a gradual recovery of the adrenal axis was seen. This case shows that acute adrenal crisis may be a consequence even at the usual prescribed doses, stressing the importance of using the lowest dose of inhaled steroids needed to control symptoms and having an increased awareness of this complication.

Early presentation of familial paraganglioma with SDHB mutation in a 13 year old child and its mother.

Paragangliomas are extradadrenal neuroendocrine tumors, recently associated with gene mutations in the succinate dehydrogenase complex (SDH). These mutations are thought to be responsible for the familial paraganglioma syndrome. Average age of tumor diagnosis for SDH mutation carriers is about 30 years of age, but patients younger than 10 years have been reported. We present the case of a 13 year-old boy with abdominal paraganglioma, whose mother also had a history of thoracic paraganglioma diagnosed at 14 years of age. Both were found to carry a mutation in exon 4 of the SDHB gene, heterozygous for c.418G>T, p.Val140Phe sequence. Compared to the other SDH subtypes, SDHB associated tumors have been found to be much more aggressive. This has led to current recommendations that tumor screening of asymptomatic SDHB carriers should start as early as 10 years of age. An even earlier initiation seems warranted for the identified carriers in this family.

Chromosome 17 polysomy without human epidermal growth factor receptor 2 amplification does not predict response to lapatinib plus paclitaxel compared with paclitaxel in metastatic breast cancer.

PURPOSE: It has been suggested that a subgroup of human epidermal growth factor receptor 2 (HER2)-negative breast cancer patients with chromosome 17 (Chr-17) polysomy benefit from HER2-directed therapy. This hypothesis was examined using the data from a phase III trial that randomized patients with HER2-negative or HER2-untested metastatic breast cancer to first-line therapy with paclitaxel along with either lapatinib or placebo. EXPERIMENTAL DESIGN: HER2 expression level by immunohistochemistry, fluorescence in situ hybridization (FISH), and mean HER2 ratio of Chr-17 values were determined centrally using archival tissue. Polysomy means of 2.0 and 2.2 served as thresholds. RESULTS: Of 580 patients on the original trial, 406 were HER2 negative by FISH. Progression-free survival (PFS) data were available for 405 patients, of whom 44 (11%) met the definition of polysomy (Chr-17 >or=2.2, FISH negative for HER2). Median PFS in the polysomy group was 20.9 and 24.4 weeks for paclitaxel plus lapatinib and paclitaxel plus placebo, respectively. In the nonpolysomy group, median PFS was 24.6 and 23.1 weeks for paclitaxel plus lapatinib and paclitaxel plus placebo, respectively. Log-rank testing showed no treatment advantage for either group. Similar results were found using a Chr-17 polysomy cutoff of 2.0. Response rates in the polysomy group were 17% for paclitaxel plus lapatinib and 10% for paclitaxel plus placebo. In the nonpolysomy group, response rates were 32% for paclitaxel plus lapatinib and 25% for paclitaxel plus placebo. Neither comparison was statistically significant. CONCLUSION: This analysis could not confirm the hypothesis that Chr-17 polysomy in HER2-nonamplified patients improved chemotherapy outcome when lapatinib is added as a HER2-targeted treatment.

Qualidade dos grãos de milho utilizados em uma indústria alimenticia de Campina Grande-PB, no período de 2004–2005 / Quality of corn grain used in a food industry of Campina Grande-PB

Grain corn is predisposed to fungal growth and biosynthesis of aflatoxins. This research has the objective of evaluating the quality of grain corns used as raw material to make corn powder, regarding the presence of fungi and aflatoxins. The achieved results were compared to humidity level and water activity. The grain corn samples were analyzed from July 2004 to July 2005. The macrobiotic research of fungi was carried out via the macro and micromorphological identification and via the aflatoxins research, using the Thin Layer Chromatography Technique (TLC) as well as Bright Greenish Yellow Fluorescence (BGYF). In order to evaluate the humidity level and the water activity (Aw), the standard greenhouse method and the static method with the NOVASINA equipment were respectively used. The 40 samples analyzed did not present the aflatoxins B1, B2, G1 or G2. Eight samples presented number of fluorescent points when the BGYF technique was used. The humidity did not exceed the limit allowed by the Brazilian legislation – 14.50 per cent. There was growth of Aspergillus sppin 100 per cent of the samples; and there was growth of A. flavus in 22.5 per cent of the samples analyzed. The grain corns analyzed follow the rules of the Brazilian legislation.

O milho é um grão susceptível ao desenvolvimento fúngico e à biossíntese das aflatoxinas. A presente pesquisa teve como objetivo avaliar a qualidade dos grãos de milho utilizados como matéria-prima para fabricação de farinha de milho, quanto à presença de fungos e aflatoxinas. Os resultados obtidos foram comparados com o teor de umidade e a atividade de água. As amostras foram analisadas no período de julho de 2004 a julho de 2005. A pesquisa da macrobiota fúngica realizou-se através de identificação macro e micromorfológica e a pesquisa de aflatoxinas empregando-se atécnica de Cromatografia em Camada Delgada (CCD) e Brigth Greenish Yellow Fluorescence (BGYF). Para a avaliação do teor de umidade e a atividade de água (Aw), utilizou-se o método padrão de estufa e o método estático com aparelho NOVASINA, respectivamente. Nas 40 amostras analisadas não foi detectada a presença das aflatoxinas B1, B2, G1 e G2. Pela técnica BGYF, 8 amostras apresentaram número de pontos fluorescentes. A umidade não excedeu o limite permitido pela legislação brasileira – 14,50 por cento. Em 100 por cento das amostras houve crescimento de Aspergillus spp.; em 22,5 por cento das amostras analisadas houve crescimento de A. flavus. Os grãos de milho analisados estão de acordo com a legislação vigente do país.

Laparoscopic excision of a familial paraganglioma.

Pheochromocytomas are rare neuroendocrine tumors that secrete catecholamines and usually arise from the adrenal medulla. Catecholamine-producing tumors that arise from extra-adrenal chromaffin tissue are referred to as paragangliomas, or extra-adrenal pheochromocytomas. Contrary to the traditional "Rule of Tens," as many as 25% of pheochromocytomas occur in hereditary tumor syndromes, such as multiple endocrine neoplasia-2, von Hippel-Lindau disease, neurofibromatosis-1, or hereditary or familial paraganglioma syndrome. Surgical resection remains the only curative therapy for pheochromocytomas, and advances in minimally invasive techniques have shown laparoscopic adrenalectomy to be safe and effective. Paragangliomas are extremely rare tumors, especially in children, and the role of laparoscopy in their treatment is evolving. This case report and the accompanying video demonstrate that the laparoscopic approach to retroperitoneal paraganglioma resection provides excellent exposure of the tumor and surrounding structures. Given proper patient preparation and perioperative monitoring, laparoscopic exploration and resection of paragangliomas can be safely and successfully accomplished in children. In addition, genetic analysis should be considered for all patients presenting with these tumors.

HER-2 gene amplification, HER-2 and epidermal growth factor receptor mRNA and protein expression, and lapatinib efficacy in women with metastatic breast cancer.

PURPOSE: Biomarkers from two randomized phase III trials were analyzed to optimize selection of patients for lapatinib therapy. EXPERIMENTAL DESIGN: In available breast cancer tissue from EGF30001 (paclitaxel +/- lapatinib in HER-2-negative/unknown metastatic breast cancer, n = 579) and EGF100151 (capecitabine +/- lapatinib in HER-2-positive metastatic breast cancer, n = 399), HER-2 gene amplification by fluorescence in situ hybridization (FISH), HER-2 mRNA by reverse transcription-PCR (RT-PCR), HER-2 protein expression by HercepTest immunohistochemistry (IHC), epidermal growth factor receptor (EGFR) mRNA level by RT-PCR, and EGFR protein by IHC were analyzed and compared with clinical outcome. HER-2 was determined by FISH in an academic reference/research laboratory and in a large, high-volume commercial reference laboratory. RESULTS: The HER-2 gene was amplified in 47% (344 of 733) and IHC was 3+ in 35% (279 of 798), with significant correlation (P < 0.01) between FISH and IHC. Positive EGFR immunostaining (IHC 1+, 2+, or 3+) in 28% (213 of 761) correlated with EGFR mRNA levels by RT-PCR (r = 0.59; P < 0.01). HER-2 gene amplification/overexpression was associated with improved clinical outcomes (progression-free survival; P < 0.001) in both trials. A significant improvement in outcome was seen in FISH-positive and IHC 0, 1+, or 2+ patients. HER-2 mRNA expression correlated with HER-2 FISH (r = 0.83) and IHC status (r = 0.72; n = 138). No correlation was found between EGFR expression (IHC or mRNA) and responsiveness to lapatinib regardless of HER-2 status. Although a significant correlation with lapatinib responsiveness was observed among "HER-2-negative" breast cancer patients in the large, high-volume commercial reference laboratory, this was not confirmed in the academic reference/research laboratory. CONCLUSIONS: Women with HER-2-positive metastatic breast cancer benefit from lapatinib, whereas women with HER-2-negative metastatic breast cancer derive no incremental benefit from lapatinib.

Diagnostic evaluation of HER-2 as a molecular target: an assessment of accuracy and reproducibility of laboratory testing in large, prospective, randomized clinical trials.

PURPOSE: To critically assess the accuracy and reproducibility of human epidermal growth factor receptor type 2 (HER-2) testing in outside/local community-based hospitals versus two centralized reference laboratories and its effect on selection of women for trastuzumab (Herceptin)-based clinical trials. EXPERIMENTAL DESIGN: Breast cancer specimens from 2,600 women were prospectively evaluated by fluorescence in situ hybridization (FISH) for entry into Breast Cancer International Research Group (BCIRG) clinical trials for HER-2-directed therapies. RESULTS: HER-2 gene amplification by FISH was observed in 657 of the 2,502 (26%) breast cancers successfully analyzed. Among 2,243 breast cancers with central laboratory immunohistochemistry (10H8-IHC) analysis, 504 (22.54%) showed overexpression (2+ or 3+). Outside/local laboratories assessed HER-2 status by immunohistochemistry in 1,536 of these cases and by FISH in 131 cases. Overall, the HER-2 alteration status determined by outside/local immunohistochemistry showed a 79% agreement rate [kappa statistic, 0.56; 95% confidence interval (95% CI), 0.52-0.60], with FISH done by the central laboratories. The agreement rate comparing BCIRG central laboratory 10H8-IHC and outside/local laboratory immunohistochemistry was 77.5% (kappa statistic, 0.51; 95% CI, 0.46-0.55). Finally, HER-2 status, determined by unspecified FISH assay methods at outside/local laboratories, showed a 92% agreement rate (kappa statistic, 0.83; 95% CI, 0.73-0.93), with FISH done at the BCIRG central laboratories. CONCLUSIONS: Compared with the HER-2 status determined at centralized BCIRG reference laboratories, these results indicate superiority of FISH to accurately and reproducibly assess tumors for the HER-2 alteration at outside/local laboratories for entry to clinical trials.