Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of nonsentinel nodes following completion lymphadenectomy for melanoma.

INTRODUCTION: Most melanoma patients with sentinel lymph nodes (SLN) that are histologically positive for metastasis have no additional positive lymph nodes found upon completion lymph node dissection (CLND). Therefore, it has been suggested that CLND may not be required for all patients with positive SLN. This study was undertaken to determine the frequency with which nonsentinel nodes contain melanoma cells detected by RT-PCR. METHODS: Negative control lymph nodes were obtained from patients with breast and colon cancer. Positive control lymph nodes contained histologic evidence of melanoma. Nonsentinel nodes were harvested from melanoma patients undergoing CLND for a positive SLN. RT-PCR analysis for melanoma markers tyrosinase, gp100, MART-1, and MAGE-3 was performed, with Southern blot detection. The RT-PCR test was considered positive for the presence of melanoma cells if tyrosinase and at least one other marker were detected above background levels. RESULTS: RT-PCR analysis detected the presence of melanoma cells in 0/100 (0%) of negative control lymph nodes and 28/29 (97%) of positive control lymph nodes. A total of 117 histologically negative nonsentinel nodes from 13 patients who underwent CLND for positive SLN were evaluated. RT-PCR analysis was positive in 18/117 histologically negative nonsentinel nodes (15%) from 7/13 patients (54%). CONCLUSION: RT-PCR analysis suggests that when the SLN contains histologic evidence of melanoma, the remaining nodes in that basin are at risk for metastatic disease, despite the fact that these nonsentinel nodes are infrequently histologically positive.

The role of the ultrasonically activated shears and vascular cutting stapler in hepatic resection.

Hemorrhage and liver failure are the two greatest concerns for patients undergoing major liver resection. Inflow occlusion (Pringle maneuver) is often used to minimize blood loss, but hepatic ischemia results in an increased risk of postoperative hepatic dysfunction. We report our experience with the Harmonic Scalpel ultrasonically activated shears (UAS; Ethicon Endo-Surgery, Cincinnati, OH) and a vascular stapler for hepatic resection as technological advances that aid in minimizing blood loss and thereby reduce the need for inflow occlusion. We retrospectively reviewed liver resections performed from September 1997 through July 1998, in which the UAS and articulating vascular endoscopic linear cutting stapler were used. The vascular stapler was used to divide the appropriate portal vein branch and hepatic vein(s) before parenchymal transection. Parenchymal dissection was performed with UAS to a depth of approximately 2 to 3 cm, and the remainder of the liver parenchyma was divided by a clamp crush and clip and suture ligate technique. Patients underwent segmental resection (n = 12), lobectomy (n = 13), or extended lobectomy (n = 11). Resection was performed for metastatic disease, primary liver tumors, or benign disease in 21, 8, and 7 patients, respectively. A Pringle maneuver was performed in 7 of 36 patients (mean clamp time, 8 minutes). The median required intraoperative blood transfusion was 0 units of packed red blood cells. Major and minor complications occurred in 12 and 3 patients, respectively. Two deaths were related to pneumonia and abdominal infection. The vascular stapler safely and securely divides portal vein branches and hepatic veins. The UAS initiates parenchymal transection with minimal blood loss. These two technologies facilitate the surgeon's aim of liver resection without blood transfusion or Pringle maneuver.

Primary squamous cell carcinoma of the breast presenting as a breast abscess.

Primary squamous cell carcinoma (SCC) of the breast is a very rare neoplasm, with only 75 cases reported in the English literature. Herein, we report four new cases and discuss the diagnostic and therapeutic challenges of this unusual tumor in a retrospective review of all cases of SCC of the breast at our institution from 1990 to 1998. Four patients with breast SCC were identified, with a mean age of 70 years. Two patients with "pure" SCC (no features of ductal carcinoma) were initially treated for breast abscess. Two other patients with features of both SCC and ductal carcinoma had skin erythema associated with an underlying mass, and infectious etiology was considered in each case. Mean tumor size was 4.9 cm. Both patients with pure SCC underwent extensive evaluation for primary tumors at other sites. Two patients developed early systemic metastasis. SCC of the breast is often diagnosed at an advanced stage and may be confused with breast abscess. For this reason, breast biopsy should be considered in cases of breast abscess. Treatment of primary SCC of the breast is similar to that of more common types of breast cancer (i.e., breast conservation is possible and lymph node dissection is recommended). Because metastasis to the breast from other primary tumor sites has been reported (lung, cervix, skin, and esophagus), patients with pure SCC should undergo evaluation to exclude this possibility.

Infiltrating ductal carcinoma of the breast: the survival impact of race.

PURPOSE: Breast cancer has a poorer prognosis among black women than among white women. This review was conducted to determine whether this disparity reflects the direct impact of race on likelihood of cure or on time to death from breast cancer or stems from the interaction of race with tumor stage and patient age. PATIENTS AND METHODS: We analyzed data from 115,838 patients with localized (node-negative) and regionally metastatic (node-positive) breast cancer from the Surveillance, Epidemiology, and End-Results (SEER) Program of the National Cancer Institute. Parametric analysis was used to determine the independent prognostic value of age, stage, and race. Linear regression and distribution analyses were also used to examine the interaction of these covariates. RESULTS: The prevalence of regionally metastatic disease, relative to localized disease, declined with increased age among white patients and those classified as "other," but remained relatively constant among black patients. Parametric analysis showed a smaller cured fraction and shorter time to death when patients with regional disease were compared with those with localized disease. A similar disparity was found when black patients were compared with those classified as white or other. CONCLUSION: Age and race have a significant association with tumor stage. In addition, our data show that race has an independent impact on the clinical course of breast cancer and diminishes both the likelihood of cure and time to death among uncured patients.

Analysis of minocycline by high-performance liquid chromatography in tissue and serum.

A sensitive and rapid reversed-phase high-performance liquid chromatography assay can be used to accurately determine serum and tissue minocycline concentrations. Minocycline is a broad spectrum tetracycline derivative with many applications. Tissue and serum samples were obtained from guinea pigs that had received either topical or intravenous minocycline. Samples were extracted using a Sep-Pak C18 cartridge and were injected into a microBondapak C18 column with an isocratic methanol mobile phase. Samples were analyzed using UV detection and produced sharp peaks with a retention time of 2.5 min. The lower limit of detection was 100 ng and drug recovery was 61%. This method greatly facilitated the analysis of minocycline while allowing for sensitivity.

Absorbable, delayed-release antibiotic beads reduce surgical wound infection.

Absorbable, polyglycolic acid (PGA) beads were evaluated as a new vehicle for local antibiotic delivery. Incisions on the dorsa of guinea pigs were contaminated with 1 x 10(8) Escherichia coli and 1 x 10(8) Staphylococcus aureus. PGA beads containing either minocycline, amikacin, or no antibiotic (placebo) were placed into these wounds and compared to animals treated with systemic minocycline or amikacin alone. The diameter of wound erythema was measured daily for 7 days. Serial blood and wound quantitative cultures were obtained, and serum and wound antibiotic concentrations were determined. Both minocycline-PGA and amikacin-PGA-treated wounds exhibited less erythema than placebo-PGA wounds (P < 0.05). All minocycline-PGA and amikacin-PGA-treated wounds healed primarily, whereas 67 per cent of placebo-PGA wounds developed purulence, dehisced, and healed secondarily. Local antibiotic delivery was more effective than systemic administration in reducing wound erythema and the number of bacteria in wound quantitative cultures. E. coli and S. aureus were quantitatively reduced (P < 0.05) in wounds of antibiotic PGA-treated animals compared to those in placebo-PGA-treated and systemic minocycline and systemic amikacin-treated animals. Measurable minocycline and amikacin concentrations were present in antibiotic-PGA-treated wounds through day 3, without detectable serum levels. Delayed-release, absorbable, antibiotic-containing PGA beads effectively prevent infection in contaminated wounds and have the advantage of not requiring vehicle removal.

Suppository administration of chemotherapeutic drugs with concomitant radiation for rectal cancer.

PURPOSE: Preoperative radiation with combined chemotherapy is effective in shrinking advanced rectal cancer locally and facilitating subsequent surgery. Suppository delivery of 5-fluorouracil is associated with less toxicity and higher rectal tissue concentrations than intravenous administration. This prompted us to evaluate suppository and intravenous administration of 5-fluorouracil and mitomycin C with concomitant radiation to determine associated toxicity. METHODS: Rectal, liver, lymph node, and lung tissue and systemic and portal blood were collected serially from male Sprague Dawley rats to determine drug concentrations following suppository or intravenous delivery of 5-fluorouracil or mitomycin C. Thirty-six animals were randomly assigned to treatment groups and received 5-fluorouracil suppositories, mitomycin C suppositories, or an equivalent intravenous dose of 5-fluorouracil or mitomycin C 30 minutes before radiation therapy. Before and 3, 6, 10, and 15 days following this treatment, blood was collected, colonoscopy was performed, and rectal tissue was harvested for histologic examination. RESULTS: Mitomycin C suppository was significantly less toxic compared with intravenous delivery, and higher rectal tissue concentrations were observed from 10 to 30 minutes (P < 0.05). Compared with intravenous 5-fluorouracil administration and radiation, 5-fluorouracil suppository and radiation resulted in additive myelosuppression at day 6 (P < 0.05) with rapid recovery. CONCLUSIONS: 5-Fluorouracil and mitomycin C suppository delivery combined with radiation causes less systemic toxicity and is more effective than intravenous administration.

HPLC analysis of 5-FU and FdUMP in tissue and serum.

Tissue and serum concentrations of 5-fluorouracil (5-FU) and 5-fluoro-2'-deoxyuridine 5' monophosphate (FdUMP) can be determined using a simple and sensitive reverse phase HPLC technique. Samples are extracted with ethyl acetate, evaporated to dryness, resuspended in the mobile phase (water/methanol [85:15]), and injected into a C18 muBondapack column (250 mm x 30 mm) with a flow rate of 1 ml/min. Both fluorescence (266 lambda Excitation and 350 lambda Emission) and UV (250 nm) are used for detection. Standard solutions of 5-FU and FdUMP produce a linear relationship (r2 = 0.985) with a detection sensitivity of 50 pg. Recovery following this extraction procedure is 78%. This analytical method is simple and convenient to use for the simultaneous detection of 5-FU and FdUMP.