MammaPrint molecular diagnostics on formalin-fixed, paraffin-embedded tissue.

MammaPrint, a prognostic 70-gene profile for early-stage breast cancer, has been available for fresh tissue. Improvements in RNA processing have enabled microarray diagnostics for formalin-fixed, paraffin-embedded (FFPE) tissue. Here, we describe method optimization, validation, and performance of MammaPrint using analyte from FFPE tissue. Laboratory procedures for enabling the assay to be run on FFPE tissue were determined using 157 samples, and the assay was established using 125 matched FFPE and fresh tissues. Validation of MammaPrint-FFPE, compared with MammaPrint-fresh, was performed on an independent series of matched tissue from five hospitals (n = 211). Reproducibility, repeatability, and precision of the FFPE assay (n = 87) was established for duplicate analysis of the same tumor, interlaboratory performance, 20-day repeat experiments, and repeated analyses over 12 months. FFPE sample processing had a success rate of 97%. The MammaPrint assay using FFPE analyte demonstrated an overall equivalence of 91.5% (95% confidence interval, 86.9% to 94.5%) between the 211 independent matched FFPE and fresh tumor samples. Precision was 97.3%, and repeatability was 97.8%, with highly reproducible results between replicate samples of the same tumor and between two laboratories (concordance, 96%). Thus, with 580 tumor samples, MammaPrint was successfully translated to FFPE tissue. The assay has high precision and reproducibility, and FFPE results are substantially equivalent to results derived from fresh tissue.

The molecular basis of cancer and the development of targeted therapy.

The sequencing of the human genome and the ability to rapidly identify genes and proteins, both normal and mutant, that are involved in tumorigenesis and malignant phenotypes, have changed the ability to understand malignant cells. Understanding and applying this information to the diagnosis and treatment of cancer are facilitated best with a multidisciplinary team. The cancer surgeon plays a pivotal role in this team. This article briefly summarizes: (1) the clinically relevant applications of molecular biology to the cancer surgeon, (2) the current understanding of the molecular basis for cancer, and (3) the current targeted agents and their clinical applications.

Melanoma vaccines.

Over the last century, vaccine studies have demonstrated that the human immune system, with appropriate help, can limit or prevent infection against otherwise lethal pathogens. Encouraged by these results, success in animal models and numerous well-documented reports of immune-mediated melanoma regression in humans, investigators developed melanoma vaccines. However, despite considerable laboratory evidence for vaccine-induced immune responses, clinical responses remain poor. Recent studies have elucidated several mechanisms that hinder or prevent the creation of successful vaccines and suggest novel approaches to overcome these barriers. Unraveling the mechanisms of autoimmunity, dendritic cell activation, regulatory T cells and Toll-like receptors will generate novel vaccines that, when used in conjunction with standard adjuvant therapies, may result in improved clinical outcomes. The objective of this review is to provide an overall summary of recent clinical trials with melanoma vaccines and highlight novel vaccine strategies to evaluate in the near future.

Penetrating gastric ulcer presenting as a subcapsular liver abscess.

This report describes a subcapsular liver abscess secondary to a penetrating gastric ulcer. The initial read on the CT scan misinterpreted the abscess cavity as an opacified loop of bowel, although it was very conspicuous on a retrospective review. A penetrating gastric ulcer was identified with esophagogastroduodenoscopy and the subcapsular liver abscess was subsequently detected using MRI. Although the conventional treatment of this condition is surgery, successful management was accomplished with a combination of percutaneous drainage, Helicobacter pylori eradication, and acid-suppressive therapy. A review of the literature is provided, including associated cases and the diagnostic modalities used in the evaluation of this condition. This case illustrates how one can arrive at the correct diagnosis with the use of multiple complementary modalities of investigation.