Increased epithelial membrane protein 2 expression in glioblastoma after treatment with bevacizumab.

BACKGROUND: Antiangiogenic therapy with bevacizumab has failed to provide substantial gains in overall survival. Epithelial membrane protein 2 (EMP2) is a cell surface protein that has been previously shown to be expressed in glioblastoma, correlate with poor survival, and regulate neoangiogenesis in cell lines. Thus, the relationship between bevacizumab and EMP2 was investigated. METHODS: Tumor samples were obtained from 12 patients with newly diagnosed glioblastoma at 2 time points: (1) during the initial surgery and (2) during a subsequent surgery following disease recurrence post-bevacizumab treatment. Clinical characteristics and survival data from these patients were collected, and tumor samples were stained for EMP2 expression. The IVY Glioblastoma Atlas Project database was used to evaluate EMP2 expression levels in 270 samples by differing histological areas of the tumor. RESULTS: Patients with high EMP2 staining at initial diagnosis had decreased progression-free and overall survival after bevacizumab (median progression-free survival 4.6 months vs 5.9 months; log-rank P = .076 and overall survival 7.7 months vs 14.4 months; log-rank P = .011). There was increased EMP2 staining in samples obtained after bevacizumab treatment in both unpaired (mean H-score 2.31 vs 1.76; P = .006) and paired analyses (mean difference 0.571; P = .019). This expression increase correlated with length of bevacizumab therapy (R 2  = 0.449; Pearson P = .024). CONCLUSIONS: Bevacizumab treatment increased EMP2 protein expression. This increase in EMP2 correlated with reduced mean survival time post-bevacizumab therapy. We hypothesize a role of EMP2 in clinical bevacizumab resistance and as a potential antiangiogenic therapeutic target in glioblastoma.

Identification of HER2 Immunohistochemistry-Negative, FISH-Amplified Breast Cancers and Their Response to Anti-HER2 Neoadjuvant Chemotherapy.

Objectives: Either immunohistochemistry (IHC) or in situ hybridization (ISH) can be used to determine human epidermal growth factor receptor 2 (HER2) status. Breast cancers (BCs) with HER2 IHC-negative (IHC-) and ISH-amplified (ISH+) results have been rarely reported but not well studied. We investigated the frequency of HER2 IHC-/ISH+ BCs and their response to anti-HER2 neoadjuvant chemotherapy (NAC). Methods: Seventeen BCs with HER2 IHC-/ISH+ results were identified from 1,107 consecutive invasive BCs (1.5%, 17/1,107). Results: Gene protein assay confirmed the original HER2 IHC and ISH results. Increased HER2 RNA level was detected in HER2 IHC-/ISH+ cases compared with HER2 IHC-/ISH- cases. Eight patients had anti-HER2 NAC; three had pathologic complete response, and five had residual tumors. Conclusions: A small percentage of patients (1.5%) showed discordant HER2 IHC and ISH results (IHC-/ISH+) and would have lost the opportunity for potentially beneficial anti-HER2-targeted therapy if only HER2 IHC testing had been used.

Resección hepática por hepatocarcinoma: estudio comparativo entre pacientes menores y mayores de 70 años / Hepatic resection in hepatocarcinoma: A comparative study between patients under and over 70 years-old

Introducción: El manejo quirúrgico agresivo de ciertos tipos de tumores es cuestionado en el paciente anciano debido a la posibilidad de tener un aumento en la morbimortalidad. Este es el caso del paciente anciano con carcinoma hepatocelular. Comparamos los resultados obtenidos con la resección hepática entre pacientes mayores y menores de 70 años de edad con hepatocarcinoma. Material y métodos Se realizaron 36 resecciones hepáticas curativas para tratar cáncer hepatocelular. Dividimos nuestra población en 2 grupos (14 pacientes < 70 años y 22 pacientes ≥ 70 años de edad) y comparamos su morbimortalidad, periodo libre de enfermedad y sobrevida global utilizando curvas de Kaplan-Meir y prueba de log rank. Además buscamos factores de mal pronóstico en la población. Resultados Ambos grupos son similares en cuanto al estado pre-operatorio. El tiempo quirúrgico, tipo de procedimiento, estancia hospitalaria y morbimortalidad fueron similares. La sobrevida global para los pacientes jóvenes y seniles a 3 y 5 años fue de 85,7 vs. 68,7% y 47,6 vs. 60% respectivamente (p=0,813). El periodo libre de enfermedad a 3 y 5 años fue de 69,3 vs. 35,2% y 39,6 vs. 23,4% respectivamente (p=0,539). El análisis multivariado reveló la enfermedad multicéntrica y la alfa-feto-proteína elevada como factores pronósticos independientes de una sobrevida libre de enfermedad y global más cortas. Conclusión Los pacientes mayores de 70 años con carcinoma hepatocelular deben ser manejados en una manera similar a pacientes más jóvenes. Hay que tomar en cuenta factores de mal pronóstico como la multicentricidad y la alfa-feto-proteína elevadas (AU)

Introduction: The ageing population raises concerns about the adequacy of aggressive surgical procedures and their outcomes. The treatment of the elderly with hepatocellular carcinoma is one of the diseases that involve complicated management decisions. We set out to compare the results between an older and younger patient cohort with this disease are compared. Material and methods: A total of 36 hepatic resections were performed on patients with hepatocellular carcinoma between 2000 and 2011. The cohort was divided into 2 groups (14patients < 70 and 22 patients 70 years of age), and their results, disease free and overall survival were compared using Kaplan-Meir curves and log rank test. An attempt was also made at determining the predictive factors of a poor outcome among this patient cohort. Results: Both groups were similar with regards to their pre-operative status. Operation time, procedure, hospital stay, and morbidity and mortality were similar. Overall survival at 3and5 years comparing the younger vs. the elderly group was 85.7% vs. 68.7% and 47.6% vs. 60%,respectively (P =.813). Disease free survival at 3 and 5 years comparing the younger vs. the elderly group was 69.3% vs. 35.2% and 39.6% vs. 23.4%, respectively (P =.539). Multivariate analysis of the whole cohort revealed multicentric diseases and elevated alpha-fetoproteinas independent factors of poor disease free survival and overall survival, respectively. Conclusions: Elderly patients with hepatocellular carcinoma should be managed in a similar fashion to younger patients. Surgeons should expect similar post-operative complications, disease free and overall survival (AU)

[Hepatic resection in hepatocarcinoma: a comparative study between patients under and over 70 years-old]. / Resección hepática por hepatocarcinoma: estudio comparativo entre pacientes menores y mayores de 70 años.

INTRODUCTION: The ageing population raises concerns about the adequacy of aggressive surgical procedures and their outcomes. The treatment of the elderly with hepatocellular carcinoma is one of the diseases that involve complicated management decisions. We set out to compare the results between an older and younger patient cohort with this disease are compared. MATERIAL AND METHODS: A total of 36 hepatic resections were performed on patients with hepatocellular carcinoma between 2000 and 2011. The cohort was divided into 2 groups (14 patients < 70 and 22 patients ≥ 70 years of age), and their results, disease free and overall survival were compared using Kaplan-Meir curves and log rank test. An attempt was also made at determining the predictive factors of a poor outcome among this patient cohort. RESULTS: Both groups were similar with regards to their pre-operative status. Operation time, procedure, hospital stay, and morbidity and mortality were similar. Overall survival at 3 and 5 years comparing the younger vs. the elderly group was 85.7% vs. 68.7% and 47.6% vs. 60%, respectively (P=.813). Disease free survival at 3 and 5 years comparing the younger vs. the elderly group was 69.3% vs. 35.2% and 39.6% vs. 23.4%, respectively (P=.539). Multivariate analysis of the whole cohort revealed multicentric diseases and elevated alpha-fetoprotein as independent factors of poor disease free survival and overall survival, respectively. CONCLUSIONS: Elderly patients with hepatocellular carcinoma should be managed in a similar fashion to younger patients. Surgeons should expect similar post-operative complications, disease free and overall survival.

Development of automated brightfield double in situ hybridization (BDISH) application for HER2 gene and chromosome 17 centromere (CEN 17) for breast carcinomas and an assay performance comparison to manual dual color HER2 fluorescence in situ hybridization (FISH).

BACKGROUND: Human epidermal growth factor receptor 2 (HER2) fluorescence in situ hybridization (FISH) is a quantitative assay for selecting breast cancer patients for trastuzumab therapy. However, current HER2 FISH procedures are labor intensive, manual methods that require skilled technologists and specialized fluorescence microscopy. Furthermore, FISH slides cannot be archived for long term storage and review. Our objective was to develop an automated brightfield double in situ hybridization (BDISH) application for HER2 gene and chromosome 17 centromere (CEN 17) and test the assay performance with dual color HER2 FISH evaluated breast carcinomas. METHODS: The BDISH assay was developed with the nick translated dinitrophenyl (DNP)-labeled HER2 DNA probe and DNP-labeled CEN 17 oligoprobe on the Ventana BenchMark(R) XT slide processing system. Detection of HER2 and CEN 17 signals was accomplished with the silver acetate, hydroquinone, and H2O2 reaction with horseradish peroxidase (HRP) and the fast red and naphthol phosphate reaction with alkaline phosphatase (AP), respectively. The BDISH specificity was optimized with formalin-fixed, paraffin-embedded xenograft tumors, MCF7 (non-amplified HER2 gene) and BT-474 (amplified HER2 gene). Then, the BDISH performance was evaluated with 94 routinely processed breast cancer tissues. Interpretation of HER2 and CEN 17 BDISH slides was conducted by 4 observers using a conventional brightfield microscope without oil immersion objectives. RESULTS: Sequential hybridization and signal detection for HER2 and CEN 17 ISH demonstrated both DNA targets in the same cells. HER2 signals were visualized as discrete black metallic silver dots while CEN 17 signals were detected as slightly larger red dots. Our study demonstrated a high consensus concordance between HER2 FISH and BDISH results of clinical breast carcinoma cases based on the historical scoring method (98.9%, Simple Kappa = 0.9736, 95% CI = 0.9222 - 1.0000) and the ASCO/CAP scoring method with the FISH equivocal cases (95.7%, Simple Kappa = 0.8993%, 95% CI = 0.8068 - 0.9919) and without the FISH equivocal cases (100%, Simple Kappa = 1.0000%, 95% CI = 1.0000 - 1.0000). CONCLUSION: Automated BDISH applications for HER2 and CEN 17 targets were successfully developed and it might be able to replace manual two-color HER2 FISH methods. The application also has the potential to be used for other gene targets. The use of BDISH technology allows the simultaneous analyses of two DNA targets within the context of tissue morphological observation.

Ammonia metabolism in Aedes aegypti.

We investigated the mechanisms by which Aedes aegypti mosquitoes are able to metabolize ammonia. When females were given access to solutions containing NH(4)Cl or to a blood meal, hemolymph glutamine and proline concentrations increased markedly, indicating that ammonium/ammonia can be removed from the body through the synthesis of these two amino acids. The importance of glutamine synthetase was shown when an inhibitor of the enzyme was added to the meal causing the glutamine concentration in hemolymph to decrease significantly, while the proline concentration increased dramatically. Unexpectedly, we found an important role for glutamate synthase. When mosquitoes were fed azaserine, an inhibitor of glutamate synthase, the glutamine concentration increased and the proline concentration decreased significantly. This confirms the presence of glutamate synthase in mosquitoes and suggests that this enzyme contributes to the production of glutamate for proline synthesis. Several key enzymes related to ammonium/ammonia metabolism showed activity in homogenates of mosquito fat body and midgut. The mosquito genes encoding glutamate dehydrogenase, glutamine synthetase, glutamate synthase, pyrroline-5-carboxylate synthase were cloned and sequenced. The mRNA expression patterns of these genes were examined by a real-time RT-PCR in fat body and midgut. The results show that female mosquitoes have evolved efficient mechanisms to detoxify large loads of ammonium/ammonia.