Negative regulation of UCP2 by TGFß signaling characterizes low and intermediate-grade primary breast cancer.

The histological manifestation of growth-regulating and differentiation-inducing signals in cancer cells is considered as a key component for clinical outcome prediction and commonly defined as tumor differentiation grade. However, the molecular and functional framework underlying this clinical parameter remains poorly understood. Our correlative data display a significant association (P>0.001) between mitochondrial uncoupling protein 2 (UCP2) and tumor grade in primary breast cancer (n=234). Through mechanistic analyses, we show a synergistic link between UCP2 and established cellular pathways in conferring grade-associated functional phenotypes. Here, the application of well to moderately differentiated primary tumor cell lines has enabled direct observation of SMAD recruitment to the UCP2 promoter underlying repression of gene transcription. In contrast, poorly differentiated tumor cells, known to be TGFß resistant, displayed aberrant UCP2 regulation, and consequently, gene overexpression, which reduced mitochondrial calcium and facilitated the maintenance of mitochondrial membrane potential, thereby significantly decreasing oxidative stress and inhibiting cell death. Conversely, UCP2 silencing in such cells rapidly led to the induction of apoptosis and cell differentiation, concurrent with reduced cell survival and proliferation, confirming gene-specific effects. Demonstration of a biologically driven role for UCP2 dysregulation in promoting multiple characteristics of tumor aggressiveness strongly endorses assessment of gene expression at clinical presentation to augment therapeutic decision-making and improve patient outcome through personalized targeting approaches.

Psychometric properties of child- and teacher-reported curl-up scores in children ages 10-12 years.

The purpose of this study was to examine the psychometric properties of child- and teacher-reported curl-up (CU) scores in children ages 10-12 years in both a norm-referenced (NR) and criterion-referenced (CR) framework. Eighty-four children, 36 boys and 48 girls, performed the FITNESSGRAM (Cooper Institute for Aerobics Research, 1992) CU test on 2 days separated by 48-72 hr. Two video cameras were used to record students' CU performances. Two students performed the CU at the same time, with each child's performance recorded by one camera. The test was terminated when the child stopped due to fatigue or after two form errors occurred. Teacher-reported scores were the average of two independent ratings of each video performance, while child-reported scores came from data collected and recorded by the children. Single trial norm-referenced reliability was R = .75 for girls and R = .80 for boys for teacher-reported CU and R = .69 and R = .70 for child-reported CU for girls and boys, respectively. CR reliability was examined using P, proportion of students who consistently passed or failed the test across 2 days, and km, defined as reliability with chance removed. For teacher-reported scores, P = .89 and km = .78 for boys and P = .81 and km = .62 for girls. For child-reported scores, P = .86 and km = .72 for boys, while P = .79 and km = .58 for girls. For teacher-reported data, 39% of boys passed and 50% failed the test on both days, while for girls the percentages were 27% pass and 54% fail. For child-reported data, 64% of boys passed and 22% failed on both days, while 54% of girls passed and 25% failed. NR validity was examined by correlating teacher and child-reported scores. The resultant coefficient was r = .42 (95% CI = .11-.66) for boys and r = .67 (95% CI = .58-.74) for girls. Additionally, child-reported scores were significantly higher than teacher-reported scores. CR validity was examined with a contingency coefficient, and results indicated C = .55 with 44% false master errors for boys and C = .65 with 29% false master errors for girls. The findings of this study suggest that while NR reliability estimates were moderate for teacher-reported scores, single trial estimates suggest that child-reported CU should be viewed with caution. In regard to CR reliability, both teacher-reported and child-reported reliability were moderate. However, there were marked differences between teacher- and child-reported scores, with children reporting higher percentages of students passing and lower percentage of student failing the test when compared with scores reported by teachers. Validity was rather moderate when viewed in either a NR and CR framework. It is suggested that problems with child-reported scores may be due to the need for additional practice or simplification of the testing protocol.

Genomic alterations in tubular breast carcinomas.

Tubular carcinoma of the breast is a well-differentiated variant of invasive ductal carcinoma and has been shown to have an exceptionally favorable prognosis, as manifested by a low incidence of lymph node metastases and an excellent overall survival. It is unknown whether this subtype represents an early step along the continuum of development to a more aggressive, poorly differentiated ductal cancer, or whether these cancers are destined to remain well differentiated with limited metastatic potential. We undertook an analysis of 18 pure tubular carcinomas of the breast using comparative genomic hybridization to evaluate the chromosomal changes in these tumors. An average of 3.6 chromosomal alterations of the genome were identified per case. The most frequent change involved loss of 16q (in 78% of tumors) and gain of 1q (in 50% of tumors). All but one case with 1q gain also exhibited a concomitant 16q loss. Other frequent changes involved 16p gain in 7 of 18 cases (39%) and distal 8p loss in 5 of 18 cases (28%). Comparison with known genomic alterations in a mixed group of invasive cancers shows tubular cancer to have fewer overall chromosomal changes per tumor (P <.01), higher frequency of 16q loss (P <.001), and lower frequency of 17p loss (P =.007). These results strongly suggest that tubular carcinomas are a genetically distinct group of breast cancers.

Comparison of P300 from passive and active tasks for auditory and visual stimuli.

The P300 event-related brain potential (ERP) was elicited with a stimulus-sequence paradigm for auditory and visual stimuli in separate active and passive task response conditions. Auditory stimuli in the passive task yielded P300 waveforms similar to those obtained from the active task condition. Visual stimuli in the passive task yielded much smaller P300 waveforms that were not morphologically consistent with those from the active task. The results suggest that auditory stimuli produce more robust P300 components than visual stimuli in passive task situations.

Medullary carcinoma is associated with expression of intercellular adhesion molecule-1. Implication to its morphology and its clinical behavior.

The histological hallmarks for the diagnosis of medullary breast cancer are circumscription, syncytial architecture, diffuse inflammatory infiltrate, and highly atypical nuclei. The biological and prognostic implication is a lower propensity to metastasize. We studied 19 medullary carcinomas for expression of the intercellular adhesion molecule-1 and lymphocyte-function-associated antigen-1, Neu differentiation factor, tumor necrosis factor-alpha, and the expression of HER-2/neu, HER-4, and HER-3 receptors. Our study revealed that all of the 19 medullary carcinomas expressed the intercellular adhesion molecule-1 and lymphocyte function associated antigen. Eighteen of 19 cancers expressed Neu differentiation factor and tumor necrosis factor-alpha. All medullary cancers expressed the HER-2/neu receptor, however, in the majority of the cases, the staining was confined to the cytoplasm. Only 4 of 12 cancers expressed HER-4 and none of the eight medullary cancers tested expressed HER-3. By comparison, in a control group of infiltrating ductal carcinomas, expression of intercellular adhesion molecule-1, lymphocyte function associated antigen-1, and Neu differentiation factor was positive in about 25 to 30% of the cases, HER-4 was expressed in 75% and HER-3 in 95% of the cases. Taken together, our observations suggest that the expression of intercellular adhesion molecule-1, lymphocyte function associated antigen, Neu differentiation factor, and tumor necrosis factor-alpha as factors that may affect the special morphology and the biological behavior that characterizes medullary carcinomas.

Protocol for the pathologic examination and tissue processing of the mammographically directed breast biopsy.

Achieving the maximum yield of breast cancers detected by mammography has required certain changes in tissue handling and examination of the mammographically directed breast biopsy. This new radiographic technique, increased use of breast-conserving surgical approaches for the treatment of breast cancer, more enlightened and demanding patients, and increasing medical-legal exposure have all contributed to changes in the way surgical pathologists should process and sample breast biopsy specimens.

Impact of mammographic screening on the size and the relative frequency of invasion in breast cancers seen in a community hospital from 1975-1988.

At CPMC routine mammographic screening was introduced in late 1975. The total volume of breast cancers, other than outside consultations, seen in the department increased from 71 in 1975 to 164 by 1988, an increase of 230%. This expansion in volume was due largely to surgical removal of mammographically detected occult, in situ duct and relatively small invasive duct carcinomas. In 1974, prior to routine mammographic screening, in situ carcinomas represented only 4% of all breast cancers seen in the Department of Pathology at CPMC. However, after the introduction of mammographic screening, the proportion of in situ cancers increased steadily. By 1988, 45% of all breast cancers seen in our hospital were found by mammography. While data on the size distribution of invasive breast cancers are not available at our hospital prior to 1976, an appreciable effect of mammography is still evident when the numbers of relatively small invasive cancers detected in 1976 are compared with those detected in 1988. Invasive breast cancers 10 mm in diameter or less represented only 6% of all cancers in our series in 1976, but 33% in 1988. These findings confirm observations made by Gibbs on the pathology of breast cancers found in mammographically screened and unscreened populations. The detection of increasing numbers of relatively small invasive duct carcinomas produced an overall reduction in the average diameters of invasive cancers seen at CPMC. The average dropped from 30 mm in 1975 to a low of 14.8 mm in 1987. Mammography did not appear to be effective in the early detection of invasive lobular cancers and had no impact on reducing their size. The implications of early discovery of in situ duct and relatively small invasive duct carcinomas are for improved patient survival through: (1) preventing progression of in situ duct to invasive duct cancers, and (2) the removal of invasive duct cancers before reaching a size where there is a high risk of metastasis.

Processing the small invasive carcinoma detected in the mammographically directed breast biopsy.

During the next few years pathologists can expect to be called upon with increasing frequency to extract data on a number of cell cycle, biochemical, and genetic features of breast cancers as a routine part of the specimen examination. Obtaining this information without compromising the histologic diagnosis or losing morphologic-based prognostic information is challenging, particularly with small invasive breast cancers, but is a service pathologists should be able to provide.

Significance of DNA content and proliferative rate of the invasive carcinoma found in the mammographically directed breast biopsy.

During the last few years considerable effort has been made to find genetic and biochemical markers that could be used to predict the behavior of cancers with greater accuracy than that obtained by histologic grading or the assessment of other established morphologic features. Because there is considerable evidence that malignant neoplasms have cytogenetic abnormalities, and because atypical features of the nucleus such as pleomorphism and hyperchromasia are known to be prognostically unfavorable findings, quantitative analysis of nuclear DNA was a prime candidate for evaluation. Also, the rate at which malignant tumor cells replicate, and the fraction of tumor cells participating in the proliferative process, are of considerable interest to oncologists for use in assessing patient prognosis and for guiding decisions on the use of various endocrine and cytotoxic agents for adjuvant endocrine and chemotherapy.

Ultrastructural and morphometric features of poorly differentiated and undifferentiated lung tumors.

Initial results are reported from a study of lung carcinomas and carcinoid tumors performed with electron microscopy and morphometric analysis of electron micrographs. The ultrastructural findings are being used to define the range of ultrastructures of the different tumor categories diagnosed by light microscopy using the revised World Health Organization classification. Morphometric data reveal a considerable degree of overlap in cell and nuclear size among different lung tumor types.

Phenamil, an amiloride analogue, inhibits differentiation of Friend murine erythroleukemic cells.

Amiloride has been reported to inhibit Friend murine erythroleukemic (MEL) cell commitment to differentiate by inhibiting the MEL cell plasma membrane Na+-Ca2+ antiporter (R. L. Smith, I. G. Macara, R. Levenson, D. Housman, and L. Cantley. J. Biol. Chem. 257: 773-780, 1982). We therefore screened a series of amiloride analogues to determine whether a more potent and specific inhibitor of MEL cell differentiation could be found. In our experiments, as in those of Lubin (J. Cell. Physiol. 124: 539-544, 1985), amiloride itself did not inhibit MEL cell differentiation. However, we did find that the amiloride analogue phenamil reversibly inhibits dimethyl sulfoxide (DMSO)-induced MEL cell commitment to differentiate with a K1/2 of 2.5-5.0 microM (in plasma clot assay). At an extracellular concentration of 15 microM, phenamil inhibits commitment to differentiate by approximately 90% in the plasma clot assay while having a minimal effect on growth. Phenamil is not metabolized but is rapidly taken up by MEL cells. Phenamil was most effective as an inhibitor when present during the first 12 h of DMSO treatment, indicating that phenamil affects the early commitment process rather than later steps involved in hemoglobin synthesis. Phenamil does not, however, inhibit the early differentiation-induced decrease in [Na+]i and the concomitant drop in the Na+-K+ pump rate. A specific binding site for phenamil is suggested because some analogues in which the phenamil structure is slightly modified are unable to inhibit differentiation.

The ultrastructure and immunocytochemistry of renal cell carcinoma.

The spectrum of ultrastructural features of a series of primary renal cell adenocarcinomas, including clear cell, granular, oncocytic, and sarcomatoid types, and a group of metastatic renal cell adeno-carcinomas has been studied. The contributions of electron microscopy and immunocytochemistry as adjuncts to light microscopy and histochemistry in the differential diagnosis of metastatic renal cell adenocarcinoma in various anatomic locations are reviewed.

Comparison of T-antigen expression in normal, premalignant, and malignant human colonic tissue using lectin and antibody immunohistochemistry.

The Thomsen-Friedenreich antigen has been implicated as a cancer-associated antigen in some human organs including the colon. Most previous studies of Thomsen-Friedenreich antigen expression in the colon used peanut agglutinin (PNA) to identify the immunodeterminant in tissues. However, evidence from other organs suggests that anti-T antibodies have specificities which differ from those of peanut lectin. To elucidate the nature of the T-immunodeterminant in colonic mucosa, we compared staining by PNA to that of a polyclonal (PAb) and monoclonal (MAb) anti-T antibody. PNA demonstrated the best sensitivity (91%) in cancer tissues but the lowest specificity (68%) in normal mucosa. Staining with MAb was only 76% sensitive but 100% specific. Sensitivity and specificity of PAb were intermediate between PNA and MAb. MAb stained fewer adenomatous polyps than either PNA or PAb, but staining appeared to correlate with premalignant features of the polyps. PNA-binding sites were more prevalent than either PAb or MAb in hyperplastic polyps. Cell cytoplasm was stained by both antibodies more often than by PNA. The majority of fetal colonic specimens stained with all three reagents suggesting that Thomsen-Friedenreich antigen may be an oncodevelopmental antigen in human colon. Differences in staining patterns in some tissues may be due to different antigenic specificities among PNA, PAb, and MAb.

Cancer and aging--pathology.

The results of research in the areas of cytogenetics, molecular biology and cancer epidemiology have recently come together to provide insights into possible mechanisms whereby the aging process may be related to without being directly responsible for carcinogenesis. While the mechanism(s) of age-associated neoplasia have not been determined, current evidence suggests that: (1) Cancer does not appear to be the inevitable consequence of aging, that is a preprogrammed genetic time bomb ticking away just waiting to go off. (2) Cancer is an age-related disease that may arise in a hereditary form--usually in children, and in a sporadic form--usually in adults. (3) Chromosomal abnormalities associated with certain forms of cancer tend to be tumor specific and appear to be related to the activities of oncogenes. Age-related factors may mediate or modulate tumor production and progression, possibly through the conversion of proto-oncogenes to oncogenes.

Metastatic islet cell tumor in von Hippel-Lindau disease.

A 56-year-old woman with many unusual manifestations of von Hippel-Lindau syndrome is described. In addition to retinal hemangioblastomas, pheochromocytoma, renal cell carcinoma, and multiple organ cysts, she had a cerebellar astrocytoma, pancreatic exocrine insufficiency, diabetes mellitus, thyrotoxicosis, and a metastatic calcitonin-secreting islet cell carcinoma. This case report documents the first example of a metastatic islet cell tumor in a patient with von Hippel-Lindau disease. The possible relationship between this disorder, the other neurocutaneous syndromes, and the multiple endocrine neoplasia syndromes is discussed.