Three-dimensional tumor visualization of invasive breast carcinomas using whole-mount serial section histopathology: implications for tumor size assessment.

PURPOSE: Linear tumor size (T-size) estimated with conventional histology informs breast cancer management. Previously we demonstrated significant differences in margin and focality estimates using conventional histology versus digital whole-mount serial sections (WMSS). Using WMSS we can measure T-size or volume. Here, we compare WMSS T-size with volume, and with T-size measured conventionally. We also compare the ellipsoid model for calculating tumor volume to direct, WMSS measurement. METHODS: Two pathologists contoured regions of invasive carcinoma and measured T-size from both WMSS and (simulated) conventional sections in 55 consecutive lumpectomy specimens. Volume was measured directly from the contours. Measurements were compared using the paired t-test or Spearman's rank-order correlation. A five-point 'border index' was devised and assigned to each case to parametrize tumor shape considering 'compactness' or cellularity. Tumor volumes calculated assuming ellipsoid geometry were compared with direct, WMSS measurements. RESULTS: WMSS reported significantly larger T-size than conventional histology in the majority of cases [61.8%, 34/55; means = (2.34 cm; 1.99 cm), p < 0.001], with a 16.4% (9/55) rate of 'upstaging'. The majority of discordances were due to undersampling. T-size and volume were strongly correlated (r = 0.838, p < 0.001). Significantly lower volume was obtained with WMSS versus ellipsoid modeling [means = (1.18 cm3; 1.45 cm3), p < 0.001]. CONCLUSIONS: Significantly larger T-size is measured with WMSS than conventionally, due primarily to undersampling in the latter. Volume and linear size are highly correlated. Diffuse tumors interspersed with normal or non-invasive elements may be sampled less extensively than more localized masses. The ellipsoid model overestimates tumor volume.

3D Pathology Volumetric Technique: A Method for Calculating Breast Tumour Volume from Whole-Mount Serial Section Images.

Tumour size, most commonly measured by maximum linear extent, remains a strong predictor of survival in breast cancer. Tumour volume, proportional to the number of tumour cells, may be a more accurate surrogate for size. We describe a novel "3D pathology volumetric technique" for lumpectomies and compare it with 2D measurements. Volume renderings and total tumour volume are computed from digitized whole-mount serial sections using custom software tools. Results are presented for two lumpectomy specimens selected for tumour features which may challenge accurate measurement of tumour burden with conventional, sampling-based pathology: (1) an infiltrative pattern admixed with normal breast elements; (2) a localized invasive mass separated from the in situ component by benign tissue. Spatial relationships between key features (tumour foci, close or involved margins) are clearly visualized in volume renderings. Invasive tumour burden can be underestimated using conventional pathology, compared to the volumetric technique (infiltrative pattern: 30% underestimation; localized mass: 3% underestimation for invasive tumour, 44% for in situ component). Tumour volume approximated from 2D measurements (i.e., maximum linear extent), assuming elliptical geometry, was seen to overestimate volume compared to the 3D volumetric calculation (by a factor of 7x for the infiltrative pattern; 1.5x for the localized invasive mass).

Whole-specimen histopathology: a method to produce whole-mount breast serial sections for 3-D digital histopathology imaging.

AIMS: To develop a method for preparing diagnostic-quality, whole-mount serial sections of breast specimens while preserving 3-D conformation. This required supporting the fresh specimen prior to breadloafing and refining the conventional tissue processing method. The overall goal is to use digital images of whole-specimen histopathology to improve the estimation of extent of disease. METHODS AND RESULTS: To maintain a 3-D conformation, the specimen is suspended in 3.5% agar at 55 degrees C. The block is sliced at 5-mm intervals. Sectioning is performed after extended fixation in 4% formaldehyde from paraformaldehyde in 0.1 m Millonig's buffer, followed by paraffin processing using a non-routine schedule and extended paraffin infiltration. Whole-mount serial breast sections are produced with features of equal or superior quality to that which can be achieved using conventional methods. The method is compatible with some immunohistochemical stains but requires further optimization for others. CONCLUSIONS: The technique is currently suitable for research applications. With the reduction in processing time achievable with microwave-assisted processing, there is the potential for its use as a routine clinical method. This tool may improve the accuracy of margin estimates and identification of multifocality in breast cancer; further evaluation is necessary.

Malignant mullerian mixed tumors of the uterine cervix: a report of nine cases of a neoplasm with morphology often different from its counterpart in the corpus.

Malignant mullerian mixed tumors (MMMTs) of the uterine cervix are rare; less than 30 cases have been reported in the literature and only 14 have been described in detail. As a result, the clinical and pathologic features of these tumors are not well characterized. The clinicopathologic features of nine cervical MMMTs (all cases referred because of problems in differential diagnosis) are reported here and the literature on the previously described cases is reviewed. The patients ranged in age from 23 to 87 (mean 65) years. The initial manifestations were mainly vaginal bleeding or spotting or, less commonly, an abnormal Pap smear. All of the patients had a cervical mass on examination. Of the eight patients for whom staging information was available, seven were stage Ib and one was stage II. Treatment in six patients was hysterectomy with lymphadenectomy in five; postoperative radiation therapy, chemotherapy, or both were given to two of these patients. The remaining three patients were treated by local excision (with lymphadenectomy in one) followed by radiation therapy, chemotherapy, or both in two. Follow-up, available for seven patients, revealed recurrent pelvic tumor in two patients at 1.6 and 3.0 years, respectively; the former patient died from tumor at 3.5 years whereas the latter was alive with tumor at 4.5 years. Another patient was well for II years but died 13 years postoperatively from colonic adenocarcinoma. Four other patients were alive with no evidence of tumor at postoperative intervals of less than 2 years. Gross examination revealed polypoid or pedunculated masses 1.1 to 10.0 cm in maximal dimension that invaded the cervical wall in 50% of the hysterectomy specimens. On microscopic examination, three tumors contained a predominant or exclusive epithelial component of basaloid carcinoma, two contained squamous cell carcinoma, and four contained adenocarcinoma (endometrioid in three and nonspecific in one). In seven tumors, the sarcomatous component was homologous, usually resembling fibrosarcoma or endometrial stromal sarcoma; in four of these tumors, myxoid change was prominent. Two tumors contained heterologous sarcomatous elements. In three patients, a pure carcinoma abutted the MMMT: an adenoid basal carcinoma in two (with a minor component of in situ and invasive squamous cell carcinoma in each) and an endometrioid endocervical adenocarcinoma in one. These findings, combined with analysis of the previously reported cases, indicate that cervical MMMTs, compared to their counterparts in the corpus, are more commonly confined to the uterus at presentation, may have a better prognosis, and frequently have a nonglandular epithelial component.

Modulation of phenytoin teratogenicity and embryonic covalent binding by acetylsalicylic acid, caffeic acid, and alpha-phenyl-N-t-butylnitrone: implications for bioactivation by prostaglandin synthetase.

Teratogenicity of the anticonvulsant drug phenytoin is thought to involve its bioactivation by cytochromes P-450 to a reactive arene oxide intermediate. We hypothesized that phenytoin also may be bioactivated to a teratogenic free radical intermediate by another enzymatic system, prostaglandin synthetase. To evaluate the teratogenic contribution of this latter pathway, an irreversible inhibitor of prostaglandin synthetase, acetylsalicylic acid (ASA), 10 mg/kg intraperitoneally (ip), was administered to pregnant CD-1 mice at 9:00 AM on Gestational Days 12 and 13, 2 hr before phenytoin, 65 mg/kg ip. Other groups were pretreated 2 hr prior to phenytoin administration with either the antioxidant caffeic acid or the free radical spin trapping agent alpha-phenyl-N-t-butylnitrone (PBN). Caffeic acid and PBN were given ip in doses that respectively were up to 1.0 to 0.05 molar equivalents to the dose of phenytoin. Dams were killed on Day 19 and the fetuses were assessed for teratologic anomalies. A similar study evaluated the effect of ASA on the in vivo covalent binding of radiolabeled phenytoin administered on Day 12, in which case dams were killed 24 hr later on Day 13. ASA pretreatment produced a 50% reduction in the incidence of fetal cleft palates induced by phenytoin (p less than 0.05), without significantly altering the incidence of resorptions or mean fetal body weight. Pretreatment with either caffeic acid or PBN resulted in dose-related decreases in the incidence of fetal cleft palates produced by phenytoin, with maximal respective reductions of 71 and 82% at the highest doses of caffeic acid and PBN (p less than 0.05). Caffeic acid and PBN also significantly reduced the incidence of fetal resorptions produced by phenytoin, but not the fetal weight loss. In viable embryos, ASA pretreatment reduced the covalent binding of phenytoin to embryonic protein by 43% (p less than 0.05). Binding of phenytoin to embryonic resorptions was equally high with and without ASA pretreatment, and within each treatment group was 3- to 10-fold higher than that in the respective placentas and associated viable embryos (p less than 0.05). These results suggest that prostaglandin synthetase may contribute to the enzymatic bioactivation of phenytoin to a teratogenic free radical intermediate.