Characterization of gene expression profiling of mouse tissues obtained during the postmortem interval.

Attempts to establish a tissue bank from autopsy samples have led to uncovering of the secrets of many diseases. Here, we examined the length of time that the RNA from postmortem tissues is available for microarray analysis and reported the gene expression profile for up- and down-regulated genes during the postmortem interval. We extracted RNA from fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) brains and livers of three different groups of mice: 1) mice immediately after death, 2) mice that were stored at room temperature for 3h after death, and 3) mice that were stored at 4°C for 18h after death, as this storage resembles the human autopsy process in Japan. The RNA quality of the brain and the liver was maintained up to 18h during the postmortem interval. Based on the microarray analysis, we selected genes that were altered by >1.3-fold or <0.77-fold and classified these genes using hierarchical cluster analysis following DAVID gene ontology analysis. These studies revealed that cytoskeleton-related genes were enriched in the set of up-regulated genes, while serine protease inhibitors were enriched in the set of down-regulated genes. Interestingly, although the RNA quality was maintained due to high RNA integrity number (RIN) values, up-regulated genes were not validated by quantitative PCR, suggesting that these genes may become fragmented or modified by an unknown mechanism. Taken together, our findings suggest that under typical autopsy conditions, gene expression profiles that reflect disease pathology can be examined by understanding comprehensive recognition of postmortem fluctuation of gene expression.

Patient surveillance after treatment for soft-tissue sarcoma.

About 1% of all cancers are soft tissue sarcomas (STS); about 60% of these occur in the extremities. Post-treatment surveillance programs are designed to identify recurrence, new primary cancers, and complications of therapy early enough to increase survival duration and quality of life. The intensity of surveillance varies among surgeons. We hypothesized that geographic factors would account for much of this variation. The 1,592 members of the Society of Surgical Oncology were surveyed regarding their personal postoperative STS surveillance strategy using standardized clinical vignettes and a questionnaire based on the vignettes. Practice patterns were analyzed by US Census Region, Metropolitan Statistical Area (MSA), and managed care organization (MCO) penetration rate, using repeated measures analysis of variance. The study end-point was surveillance intensity. Mean follow-up intensity for the 12 surveillance modalities on the questionnaire was highly correlated with tumor size, grade, and year post surgery. Controlling for tumor stage, grade, and year post surgery, the practice location of the surgeon infrequently impacted surveillance intensity. MSA was a significant (p<0.05) predictor only of office visit frequency. MCO penetration rate significantly predicted only the frequency of urinalysis and tumor-site MRI. US Census Region significantly predicted only the frequency of LFTs. Geographic factors do not generally predict self-reported surveillance practice patterns for patients after curative-intent STS surgery. The overall variation in follow-up intensity appears to reflect factors not evaluated, such as the absence of high-quality evidence supporting any particular strategy and the quality of patients' insurance.

Multiple calcifying fibrous pseudotumors disseminated in the pleura.

Calcifying fibrous pseudotumor is an extremely rare benign lesion that develops in the pleura. We describe a case of multiple lesions in the pleura. The patient is a 52-year-old woman who had a subpleural mass on chest roentgenogram. The diagnosis was established by percutaneous needle biopsy. The largest tumor was hanging down from the parietal pleura, and additional small nodules were disseminated throughout the pleural cavity. Only a few tumors arising from the largest one were resected, and the others were left unresected. The resected tumors consisted of collagenous fibrous tissue, calcifications, and spindle cells that were positive only for vimentin immunostaining.

Metastasis of the gastrointestinal tract: FDG-PET imaging.

We assess the usefulness of F-18-fluoro-deoxyglucose (FDG) positron emission tomography (PET) in the evaluation of gastrointestinal metastases. Four cases (five lesions) in which metastases from three lung cancers and one malignant fibrous histiocytoma (MFH) of the femur were found in the gastrointestinal tract were reviewed (men/women 3 : 1, age 63-78 years, mean 72 years). The five lesions were duodenal, jejunal metastasis, and two stomach metastases from lung carcinoma, and rectal metastasis from MFH of the femur. FDG-PET was unable to detect small masses, but it was able to detect unforeseen lesions such as gastrointestinal metastases because FDG-PET is a whole-body scan in a single-operation examination. FDG-PET imaging provided valuable information for the diagnosis of gastrointestinal metastasis.

Long-term management of patients after potentially curative treatment of extremity soft tissue sarcoma: practice patterns of members of the Society of Surgical Oncology.

The optimal strategy for follow-up of extremity soft tissue sarcoma patients after primary treatment with curative intent is not known. The 1592 members of the Society of Surgical Oncology (SSO) were sent a survey to determine their practices. Office visits and chest X-rays were the most frequently performed items for all years. Complete blood count and liver function tests were the most commonly ordered blood tests, but many respondents did not order any blood tests routinely. Imaging studies of the extremities were recommended for most patients during the first postoperative year. Postoperative surveillance strategies after sarcoma treatment vary appreciably among surgeons. Surgeon age, tumor size, tumor grade, and geographic location of the surgeon account for only a modest amount of the variation, suggesting that lack of evidence supporting any particular strategy is the factor responsible for most of the variation. Efforts to provide high-quality evidence on this topic are warranted.

Extremity soft tissue sarcoma patient follow-up: tumor grade and size affect surveillance strategies after potentially curative surgery.

A large recent survey disclosed considerable variation in the intensity of follow-up after potentially curative surgery for extremity soft tissue sarcoma among highly credentialed clinicians. The source of the variation is not known. The survey was reanalyzed to investigate whether tumor grade and size account for the variation. The 1,592 members of the Society of Surgical Oncology were surveyed to measure their follow-up practices for 10 years after initial therapy for extremity sarcoma. Repeated-measures analysis of variance was used to measure the relationship among follow-up practice patterns and tumor grade, tumor size, and years after surgery. The 318 respondents who performed surgery and also carried out long-term follow-up for their patients with extremity soft tissue sarcoma were considered evaluable; 94% were general surgeons and 5% were orthopedic surgeons. Office visit, complete blood count, liver function tests, chest X-ray, chest computed tomography, extremity computed tomography, and extremity magnetic resonance imaging were ordered significantly more frequently with increasing tumor grade and size. Tumor grade and size significantly impacted physician practice patterns in post-treatment follow-up, although the degree of variation attributable to these variables was modest. These data permit rational design of a randomized clinical trial of two alternative follow-up plans.

How surgeon age affects surveillance strategies for extremity soft tissue sarcoma patients after potentially curative treatment.

BACKGROUND: The optimal strategy for follow-up of extremity soft tissue sarcoma patients after potentially curative treatment remains unknown. We investigated whether the date of completion of formal surgical training affects choice of surveillance strategy. MATERIALS AND METHODS: The 1,592 members of the Society of Surgical Oncology were asked how often they use 12 separate surveillance modalities during years 1-5 and 10 postsurgery. The motivation underlying follow-up was assessed separately. Repeated-measures analysis of variance was used to compare practice patterns by the year in which the surgeon's formal surgery training was completed, controlling for tumor grade, tumor size, and year postsurgery. RESULTS: Of the 716 respondents, 318 performed surgery and also provided long-term postoperative surveillance for their patients. These respondents were considered evaluable. Erythrocyte sedimentation rate, extremity X ray, and bone scan were the follow-up tests which differed significantly among physician age groups. Surgeons who completed training more than 30 years ago ordered erythrocyte sedimentation rate more frequently (P < 0.001). Surgeons in the 21-30 year category ordered extremity X ray and bone scan more frequently (P < 0.05), but the absolute differences among age groups were quite small. Older surgeons were also significantly more likely to believe that follow-up is clinically worthwhile. CONCLUSIONS: The posttreatment surveillance practice patterns of the members of the Society of Surgical Oncology caring for extremity soft tissue sarcoma patients vary only marginally with the length of time since completion of training. Postgraduate education may be one factor homogenizing surgeon behavior in this important aspect of cancer patient care.

Expression of glial cell line-derived neurotrophic factor correlates with perineural invasion of bile duct carcinoma.

BACKGROUND: Perineural invasion is one of the important prognostic factors for patients with bile duct carcinoma, and extensive surgery has not always improved their prognosis. It is necessary, therefore, to investigate not only clinicopathologic characteristics but also molecular mechanisms in patients with perineural invasion. The authors studied the relation between perineural invasion in patients with bile duct carcinoma and the expression of glial cell line-derived neurotrophic factor (GDNF), GDNF family receptor alpha1 (GFRalpha1), and RET receptor tyrosine kinase, which are expressed in both central and peripheral nerve tissues. METHODS: Immunohistochemical staining of GDNF, GFRalpha1, and RET was performed in 58 paraffin embedded tissue sections, including 38 sections from patients with bile duct carcinoma with perineural invasion and 20 sections from patients with bile duct carcinoma without perineural invasion. The migration of cells that expressed GDNF was analyzed by cocultivation with cells that expressed both RET and GFRalpha1. RESULTS: Moderate to strong staining of GDNF in tumor cells was observed more frequently in the sections with perineural invasion compared with the sections without invasion (P < 0.05), whereas GFRalpha1 expression in the same sections was not correlated with perineural invasion. RET expression was undetectable in specimens of bile duct carcinoma. Conversely, RET and GFRalpha1 expression were detected consistently in peripheral nerve tissues. An in vitro cell migration assay revealed that the migration of cells that expressed GDNF was enhanced by cocultivation with cells that expressed RET and GFRalpha1. The cell migration was also enhanced by the conditioned media from GDNF-treated cells that expressed RET and GFRalpha1. CONCLUSIONS: The results suggest that GDNF expression in tumor cells and GFRalpha1 and RET expression in peripheral nerve tissues may play a role in perineural invasion in patients with bile duct carcinoma through chemoattraction among these molecules.