Preservation of RNA for functional genomic studies: a multidisciplinary tumor bank protocol.

Few human tumors are collected such that RNA is preserved for molecular analysis. Completion of the Human Genome Project will soon result in the identification of more than 100,000 new genes. Consequently, increasing attention is being diverted to identifying the function of these newly described genes. Here we describe a multidisciplinary tumor bank procurement protocol that preserves both the integrity of tissue for pathologic diagnosis, and the RNA for molecular analyses. Freshly excised normal skin was obtained from five patients undergoing wound reconstruction following Mohs micrographic surgery for cutaneous neoplasia. Tissues treated for 24 hours with RNAlater were compared histologically and immunohistochemically to tissues not treated with RNAlater. Immunohistochemical stains studied included: CD45, CEA, cytokeratin AE1/3, vimentin, S-100, and CD34 on formalin-fixed, paraffin embedded tissue and CD45 staining of frozen tissue. Slides were blinded and evaluated independently by three pathologists. The histologic and immunohistochemical parameters of tissue stored in RNAlater were indistinguishable from tissue processed in standard fashion with the exception of S-100 stain which failed to identify melanocytes or Langerhan's cells within the epidermis in any of the RNAlater-treated tissues. Interestingly, nerve trunks within the dermis stained appropriately for S-100. Multiple non-cutaneous autopsy tissues were treated with RNAlater, formalin, liquid nitrogen (LN2), and TRIzol Reagent. The pathologists were unable to distinguish between tissues treated with RNAlater, formalin, or frozen in LN2, but could easily distinguish tissues treated with TRIzol Reagent because of extensive cytolysis. RNA was isolated from a portion of the tissue treated with RNAlater and used for molecular studies including Northern blotting and microarray analysis. RNA was adequate for Northern blot analysis and mRNA purified from RNAlater-treated tissues consistently provided excellent templates for reverse transcription and subsequent microarray analysis. We conclude that tissues treated with RNAlater before routine processing are indistinguishable histologically and immunohistochemically from tissues processed in routine fashion and that the RNA isolated from these tissues is of high quality and can be used for molecular studies. Based on this study, we developed a multidisciplinary tumor bank procurement protocol in which fresh tissue from resection specimens are routinely stored in RNAlater at the time of preliminary dissection. Thus, precious human tissue can be utilized for functional genomic studies without compromising the tissue's diagnostic and prognostic qualities.

Vulvar melanoma in childhood.

BACKGROUND: Malignant melanoma is rarely diagnosed during childhood. Approximately 2% of malignant melanomas occur in patients younger than 20 years, with 0.3% to 0.5% of cases occurring in prepubescent children. In adult females malignant melanoma of the vulva and vagina is 100-fold less common than malignant melanoma of nongenital skin. Malignant melanoma of the vulva occurring in a child has been reported once before. OBSERVATIONS: We report 2 cases of childhood vulvar malignant melanoma presenting in preteenage girls. In both cases, the lesions were asymptomatic enlarging hyperpigmented macules on the labium minus. In addition to features diagnostic of malignant melanoma, histological evidence of lichen sclerosus et atrophicus was identified in both lesions. Local excision with conservative margins was the treatment modality of choice in both cases, with good preservation of anatomic structure and function. CONCLUSIONS: This report is of 2 cases of vulvar melanoma in childhood, a rare, yet potentially devastating, presentation of melanoma. Biopsies on suspicious pigmented lesions on the vulva of prepubescent children should be done to rule out malignant change.

Intraoperative radiation therapy and Mohs micrographic surgery on an outpatient basis.

BACKGROUND: Intraoperative radiation therapy (IORT) is used to deliver therapeutic doses of radiation to a surgically exposed tumor or tumor bed while minimizing the radiation dose to adjacent normal tissue. It is traditionally given with the patient under general anesthesia. OBJECTIVE: To report a case of a recurrent squamous cell carcinoma treated with Mohs micrographic surgery and IORT on an outpatient basis. METHODS: Mohs surgery was used to clear the tumor in all fields except for the area of the spinal accessory nerve, which was preserved. IORT was then delivered to the area of nerve with possible residual tumor. RESULTS: The patient remains clinically tumor free 42 months posttreatment. There were no complications. CONCLUSION: Intraoperative radiation therapy can be effectively used in the outpatient setting as an adjunctive therapy after Mohs micrographic surgery.

Facial cutaneous reconstructive surgery: facial flaps.

Basic principles of aesthetics and function important for successful facial cutaneous reconstructive surgery with local flaps include the use of appropriate tissue donor sources, knowledge of the mechanisms, dynamics, and effects of tissue movement, and techniques for scar camouflage. Useful flaps for defects in each of the facial cosmetic units are reviewed with these principles used as guidelines.

Facial cutaneous reconstructive surgery: general aesthetic principles.

The performance of cutaneous reconstructive surgery requires understanding and application of many important principles. This article reviews the critical factors to consider in the management of surgical wounds by second-intention healing, primary closure, skin grafting, and repair with local flaps. For certain defects, reconstruction with local flaps offers several advantages over other alternatives. Key concepts useful in flap choice and implementation are discussed, and surgical techniques that maximize the aesthetic outcome of reconstructive surgery are reviewed.