Prognostic value of DNA flow cytometry in sympathoadrenal paragangliomas.

OBJECTIVE: To determine whether ploidy patterns are related to prognosis in sympathoadrenal paragangliomas (SAP) using flow cytometry. STUDY DESIGN: DNA flow cytometric analysis of formalin-fixed, paraffin-embedded tumor samples from 36 patients with SAP was performed. Eight cases fulfilled at least one of the following malignancy criteria: (1) extensive invasion of adjacent structures (5 cases), (2) local recurrence (3 cases), or (3) metastases (4 cases). RESULTS: Of the 36 tumors, 22 (61%) showed nondiploid patterns (12 aneuploid, 10 tetraploid). All diploid tumors were benign, while all malignant cases showed nondiploid patterns (P = .0131). The differences between diploid and aneuploid tumors and between diploid and tetraploid tumors, with regard to the malignancy of the disease, were statistically significant (P = .03311 and .01976, respectively). Only one malignant tumor had a DNA index < 1.75 (P = .00259). CONCLUSION: Anomalous DNA ploidy patterns are frequent in SAP, without necessarily implying malignancy. However, diploid DNA content may be a marker of a good prognosis. The likelihood of malignancy is greater in the tetraploid and peritetraploid range.

[DNA ploidy determination with flow cytometry, Ki-67 index and overexpression of p53 protein in 121 T1 superficial bladder carcinomas. Retrospective studies. Part II: Prognostic value and usefulness in the indication for prophylactic treatment with BCG]. / Determinación de ploidía de ADN mediante citometría de flujo, indice Ki-67 y sobre-expresión de proteína p53 en 121 carcinomas superficiales de vejiga T1. Estudio retrospectivo. 2a Parte: Valor pronóstico y utilidad en la indicación de tratamiento profiláctico con BCG.

OBJECTIVE: Evaluate the utility of Ki-67 label index, p53 expression and flow cytometry-DNA ploidy in the selection of groups to be treated with prophylactic BCG and the prognostic value compared with the classic variables (grade, lymphatic permeation, multiplicity, volume, primary). MATERIAL & METHOD: 121 superficial bladder tumors T1. 10% Cut-off level for Ki-67 and p53. Aneuplody is defined as a tumor with DNA index different of 1 or more than 20% in G2-M phase. 71 (58.7%) received BCG. RESULTS: In uni and multivariate analysis positivity to Ki-67 is correlated with recurrence. Progression is correlated with lymphatic permeation (p .0003), volume (p .016), ploidy (p .022) and positivity to p53 (p .007). In multivariate analysis, volume and positivity to p53 are independent variables. None were of utility to prevent recurrence, but Ki-67 positive or aneuploid treated tumors had less progression (p .025 and p .009 respectively). The p53 negative treated tumors had less progression too. CONCLUSIONS: Only Ki-67 is correlated with tumoral recurrence. P53 and tumor volume are correlated with stage progression. If the results are confirmed with bigger series, the Ki-67 positive and/or aneuploid tumors would obtain benefits of prophylactic treatment with BCG.

[DNA ploidy determination with flow cytometry, Ki-67 index, and overexpression of p53 protein in 121 T1 superficial bladder carcinoma. Retrospective study. Correlation with classic variables]. / Determinación de ploidía de ADN mediante citometría de flujo, índice Ki-67 y sobreexpresión de proteína P53 en 121 carcinomas superficiales de vejiga T1. Estudio retrospectivo. Correlación con las variables clásicas.

OBJECTIVE: Observe the correlation between Ki-67 label index, p53 expression and flow cytometry-DNA ploidy with the classic variables (grade, lymphatic permeation, multiplicity, volume, primary). MATERIAL AND METHOD: 121 superficial bladder tumors T1. 10% Cut-off level for Ki-67 and p53. Aneuploidy is defined as a tumor with DNA index different of 1 or more than 20% in G2-M phase. RESULTS: Statistical correlation with grade and lymphatic permeation. Ki-67 label index and p53 expression can distinguish between G1, G2 vs G3 and Lx, L0 vs. L1. The volume correlates with positivity to p53. CONCLUSIONS: Aneuploidy and positivity to Ki-67 and p53 increase with grade and lymphatic permeation.

cDNA sequence and genomic structure of the rat RET proto-oncogene.

The RET proto-oncogene, a member of the Receptor Tyrosine Kinase family, plays a crucial role during the development of the excretory system and the enteric nervous system, as demonstrated by in vivo animal studies and by its involvement in the pathogenesis of several human neurocristopathies like Hirschsprung disease and Multiple Endocrine Neoplasia type 2. Using a multistep RT-PCR approach we have isolated and sequenced the cDNA of the whole rat RET proto-oncogene, reporting the deduced amino acid sequence in comparison with the human and mouse counterparts. Moreover, two different isoforms (RET9 and RET51) have been confirmed in the rat, while a third RET isoform demonstrated in human (RET43) has not resulted to be conserved in this species. Finally, we have determined the genomic structure of the rat RET proto-oncogene comparing the exon-intron boundaries and intron sizes with the known structure of the human homologous gene. Our findings will facilitate the molecular study of appropriate rat models of RET related human diseases.

Value of ploidy pattern and nuclear texture in the diagnosis of primary hyperparathyroidism.

OBJECTIVE: To determine if there are subvisual differences (quantitation of DNA and study of chromatin texture) that distinguish single-gland parathyroid disease (adenoma) from multiple-gland disease (hyperplasia). STUDY DESIGN: Forty-eight parathyroid glands from 41 patients with primary hyperparathyroidism were studied. Cytometric differences between the chief cells and clear cells were sought. An image cytometer was used to study histologic sections stained with DNA stain reagent. In each case a total of 200 cells were measured in consecutive fields. To distinguish chief cells from clear cells, a sample of 15,600 cells collected in consecutive fields in 78 histologic sections was analyzed. RESULTS: The results indicated that none of the continuous variables differentiated between single-gland and multiple-gland parathyroid disease. The most common ploidy pattern was diploid (25/45), followed by hypodiploid (7/45) and polyploid (5/45). A tetraploid population > 3% was found in 18/45 of the cases and was more frequent in single-gland lesions (15/34) than in multiple-gland lesions (3/11). The most significant discrete variables were growth pattern and binucleation or multinucleation. Clear and chief cells were classified correctly in 61.3% of the crossover validation tests using the standard deviation of nuclear shape karyometric variable and in 76.3% using the standard deviation of maximal correlation coefficient karyometric variable. Although ploidy pattern did not show significant differences, polyploidy, aneuploidy and tetraploidy > 3% were slightly more common in chief cells. CONCLUSION: In primary hyperparathyroidism there are no subvisual differences that can distinguish adenoma from hyperplasia. Nonetheless, by cytometry, most parathyroid cells can be classified correctly as clear or chief cells.