CDHI promoter hypermethylation and E-cadherin protein expression in infiltrating breast cancer

Background: the E-cadherin gene (CDH1) maps, at chromosome 16q22.1, a region often associated with loss of heterozygosity (LOH) in human breast cancer. LOH at this site is thought to lead to loss of function of this tumor suppressor gene and was correlated with decreased disease-free survival, poor prognosis, and metastasis. Differential CpG island methylation in the promoter region of the CDH1 gene might be an alternative way for the loss of expression and function of E-cadherin, leading to loss of tissue integrity, an essential step in tumor progression.Methods: the aim of our study was to assess, by Methylation-Specific Polymerase Chain Reaction (MSP), the methylation pattern of the CDH1 gene and its possible correlation with the expression of E-cadherin and other standard immunohistochemical parameters (Her-2, ER, PgR, p53, and K-67) in a series of 79 primary breast cancers ( 71 infiltrating ductal, 5 infiltrating lobular, 1 metaplastic, 1 apocrine, and 1 papillary carcinoma).Results: CDH1 hypermethylation was observed in 72% of the cases including 52/71 ductal, 4/5 lobular carcinomas and 1 apocrine carcinoma. Reduced levels of E-cadherin protein were observed in 85% of our samples. Although not statistically significant, the levels of E-cadherin expression tended to diminish with the CDH1 promoter region methylation. In the group of 71 ductal cancinomas, most of the cases of showing CDH1 hypermethylation also presented reduced levels of expression of ER and PgR proteins, and a possible association was observed between CDH1 methylation and ER expression ( p = 0.0301, Fisher's exact test). However, this finding was not considered significant after Bonferroni correction of p-value.Conclusion: Our preliminary findings suggested that abnormal CDH1 methylation occurs in high frequencies in infiltrating breast cancers associated with a decrease in E-cadherin expression in a subgroup of cases characterized by loss of expression of other important genes to the mammary carcinogenesis process, probably due to the disruption of the mechanism of maintenance of DNA methylation in tumoral cells.

Polymorphisms of CYP17A1, CYP19, and androgen in Brazilian women with uterine leiomyomas

Background: Uterine leiomyomas are common, benign, smooth muscle tumors representing a significant public health problem. The aim of this study was to investigate CYP17A1, CYP19, and androgen (AR) polymorphisms, their relative risks for uterine leiomyomas and possible associations with clinical parameters.Methods: Uterine leiomyoma tissues and blood samples were obtained from 87 patients, as were peripheral blood samples from 68 control women. Clinical data were recorded in both groups. The CYP17A1 (rs743572) polymorphism was analyzed by PCR-RFLP, and the CYP19 [TTTA](n) repeat and AR [CAG](n) repeat were analyzed using PCR-based GeneScan analysis. AR loss of heterozygosity (LOH) and microsatellite instability were also evaluated, while samples exhibiting LOH were analyzed for X inactivation.Results: Clinical parameters related to disease development did not differ between cases and controls. CYP17A1 *A2/*A2 genotype was prevalent in non-white women. CYP17A1, CYP19, and AR genotypes and alleles did not differ between groups. However, alleles presenting [TTTA](7) repeats in intron 4 of CYP19 were more frequent in the control group (p=0.0550). Shorter and longer [CAG]n repeat alleles of AR were exclusive to the leiomyoma group. The LOH assay showed allele losses at AR locus in four informative tumors and X chromosome inactivation analysis revealed that these tumors retained the active allele.Conclusions: The overall lack of association between uterine leiomyomas with polymorphisms involved in steroidogenesis or steroid metabolism is consistent with the hypothesis that these polymorphisms do not substantially contribute to the development of these tumors.

Genetic diversity of three size classes of seeds of Eucalyptus globulus ssp. globulus

Variation in seed size is often observed in samples of eucalypt seeds and this leads to heterogeneous populations of plants, principally through variation in the early stages of plant development. It follows that samples of seeds more uniform in size could produce more uniform populations of plants. In studies of Eucalyptus globulus ssp. globulus it was of interest to determine whether or not the genetic diversity within a population, through the use of isozyme markers, was altered in the subpopulations developed from seeds of different size classes. A commercial sample of seed was separated by seed size into three subpopulations and the percentage germination and mean fresh weight of the seedlings were determined. Proteins extracted from leaves of the seedlings were separated by electrophoresis and tested for activity of eight different enzymes. These eight enzymes showed activity at 20 loci and mean genetic diversity and fixation index were determined using 13 of these loci. The subpopulation of the smallest seeds contained a greater proportion of abnormal seeds and had a lower percentage germination and plant weight compared to the other subpopulations. No significant differences were found in the number of alleles per locus, percentage of polymorphic loci, mean heterozygosity. The major part of the endogamy, indicated by F statistic, was found within the subpopulations: F-(IS) = 0.518; F-(ST) = 0.010 and F(IT) = 0.523. We conclude that the use of seeds of uniform size will lead to more uniform germination and plant growth without alteration in overall genetic diversity.

Isozyme characterization of Eucalyptus urophylla (S.T. Blake) and E-grandis (Hill ex Maiden) populations in Brazil

Seedling taken from 2 species of Eucalyptus growing in Brazil were electrophoretically analysed at 14 isozyme loci representing 6 enzyme systems: alpha-EST, beta-EST, SKDH, IDH, MDH, and LAP. Genetic variability measures were determined using 11 putative isozyme loci. on average, 81.8% and 54.5% of the loci were found to be polymorphic by the criterion of 95% in E. urophylla and E. grandis, respectively. The mean number of alleles per loci was 3.0 in E. urophylla and 2.5 in E. grandis. Observed mean heterozygosity was 0.283 in E. urophylla and 0.166 in E. grandis. Levels of genetic diversity in these species were similar to those in other Eucalyptus species which have widespread distributions. The possible hybridization of E. urophylla with E. alba is also discussed.

Chromosome 22q a frequent site of allele loss in head and neck carcinoma

Background. Loss of heterozygosity (LOH) correlates with inactivated tumor suppressor genes. LOH at chromosome arm 22q has been found in a variety of human neoplasms, suggesting that this region contains a tumor suppressor gene(s) other than NF2 important to tumorigenesis. The aim of this study was to evaluate the presence of LOH on chromosome 22q11.2-13 and determine whether there was a relationship between loss in this genomic region and tumor histologic parameters, anatomic site, and survival in patients with squamous cell carcinoma of the head and neck (HNSCC).Methods. Fifty matched blood and HNSCC tumor samples taken at the time of surgical treatment were evaluated for LOH by use of four microsatellite markers mapping to 22q11.2-q13. Clinical information was available for all patients. The frequency and distribution of LOH was correlated with clinical (age, sex, use of tobacco and alcohol, site of primary tumor, clinical stage, adjuvant therapy and overall survival) and histologic parameters (histopathologic stage, tumor differentiation).Results. LOH at 22q was found in 19 of 50 (38%) informative tumors. The respective incidence of allelic loss for the patients was as follows: 28% at D22S421, 10% at D22S277, 8% at D22S44S, and 4% at D22S280. No statistical differences were apparent with a mean follow-up of 30 months. Laryngeal tumors showed a higher incidence of LOH compared with oral tumors.Conclusions. These results suggest that the D22S277 locus may be closely linked to a tumor suppressor gene (TSG) and involved in upper aerodigestive tract carcinogenesis. In particular, laryngeal tumors may harbor another putative TSG on 22q11.2-q12.3 that may play a role in aggressive stage III/IV disease. (C) 2000 John Wiley & Sons, Inc.

Isoenzymatic variation in the germplasm of Brazilian races of maize (Zea mays L.)

Existem mais de 200 raças de milho (Zea mays L.), as quais são divididas em três grupos (raças comerciais antigas, raças comerciais recentes e raças indígenas). As raças indígenas, embora não tenham valor comercial, possuem muitas características importantes que podem ser utilizadas em programas de melhoramento de milho. A maior parte do germoplasma brasileiro das raças de milho indígena foi coletada, no mínimo, 40 anos atrás e nada é conhecido sobre a variabilidade presente neste germoplasma. Quinze populações de 4 raças indígenas de milho (Caingang, Entrelaçado, Lenha e Moroti) e 5 cultivares indígenas foram analisados utilizando-se 5 sistemas isoenzimáticos codificados por 14 locos. A análise revelou um baixo nível de variabilidade entre as amostras estudadas. O número médio de alelos/loco foi três, com 64,3% de locos polimórficos e uma heterozigosidade média esperada de 0,352. Por população, a média de número de alelos por loco polimórfico foi 1,6, em média 47,5% dos locos foram polimórficos e a heterozigosidade média foi 0,195. A distância genética média entre as populações foi 0,821 e a proporção da variabilidade genética, que é atribuída ao componente entre populações (Gst), foi 0,156. Os dados sugerem que um efeito de fundador poderia explicar a baixa variabilidade detectada.

Allozyme differentiation of two populations of the genus Neoplecostomus Eigenmann & Eigenmann, 1888 (Teleostei, Loricariidae) from the upper Paraná River basin, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genetic diversity in section Rhizomatosae of the genus Arachis (Fabaceae) based on microsatellite markers

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Isolation and characterization of microsatellite DNA markers for spinner dolphin (Stenella longirostris)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Search for Genomic Alterations in Monozygotic Twins Discordant for Cleft Lip and/or Palate

Phenotypically discordant monozygotic twins offer the possibility of gene discovery through delineation of molecular abnormalities in one member of the twin pair. One proposed mechanism of discordance is postzygotically occurring genomic alterations resulting from mitotic recombination and other somatic changes. Detection of altered genomic fragments can reveal candidate gene loci that can be verified through additional analyses. We investigated this hypothesis using array comparative genomic hybridization; the 50K and 250K Affymetrix GeneChip (R) SNP arrays and an Illumina custom array consisting of 1,536 SNPs, to scan for genomic alterations in a sample of monozygotic twin pairs with discordant cleft lip and/or palate phenotypes. Paired analysis for deletions, amplifications and loss of heterozygosity, along with sequence verification of SNPs with discordant genotype calls did not reveal any genomic discordance between twin pairs in lymphocyte DNA samples. Our results demonstrate that postzygotic genomic alterations are not a common cause of monozygotic twin discordance for isolated cleft lip and/or palate. However, rare or balanced genomic alterations, tissue-specific events and small aberrations beyond the detection level of our experimental approach cannot be ruled out. The stability of genomes we observed in our study samples also suggests that detection of discordant events in other monozygotic twin pairs would be remarkable and of potential disease significance.

Biochemical evidence of a possible new species of Neoplecostomus (Teleostei : Loricariidae) from the upper Rio Parana basin, Brazil

Neoplecostomus paranensis Langeam, 1990, from the upper Rio Parana, is the only Neoplecostomus species described in this basin and is distinguished from its congeners by the lack or reduction of the adipose fin. Neoplecostomus specimens with a normal and always present adipose fin were caught in the Rio Corumba, upper Rio Parana basin. In the present study two samples of populations, one from a tributary of Rio Paranapanema (identified as a typical N. paranensis) and the other from the Rio Corumba were compared through allozyme electrophoresis. Six diagnostic loci were found, Acp-A, Adh-A, Est-A, Gpi-A, Ldh-A and Ldh-B. In addition, the locus Gpi-B showed significant differences between allelic frequencies for the two samples. Nei's genetic identity between the populations was 0.731. The expressive genetic divergence together with the presence of an adipose fin show that the sample from the Rio Corumba is distinct from N. paranensis and probably represents a new species. (C) 2003 Published by Elsevier Ltd.

Isolation and characterization of eight microsatellite loci in Leporinus macrocephalus (Characiformes : Anostomidae) and cross-species amplification

Leporinus macrocephalus is a species endemic to the Paraguay River basin and an important fishery resource, as well as a valuable species in aquaculture programs. A total of eight polymorphic microsatellite loci were isolated and characterized. A population survey was conducted involving 45 specimens whereby a large number of alleles (range 5-17 among loci), a highly observed (0.1667-0.6129) and an expected (0.6967-0.9448) heterozygosity was detected, indicating its usefulness in population genetics studies. Cross-species amplification was successful in eight Characiformes species.

Cell death evaluation in benzo[a]pyrene-transformed human breast epithelial cells after microcell-mediated transfer of chromosomes 11 and 17

The incidence of apoptosis and nuclear instability, including the incidence of catastrophic death, were investigated in benzo[a]pyrene (BP)-transformed human breast epithelial cells (BP1-E cell line) after microcell-mediated transfer of chromosomes 11 and 17. Since the introduction of normal chromosomes 11 and 17 into tumorigenic human breast BP1-E cells reverts some of these cells' characteristics (especially those affected by microsatellite instabilities and loss of heterozygosity) those of parental non-transformed MCF-10F cells, it was expected that the cell death rates would also be affected by this treatment. The transfer of the mentioned chromosomes, especially chromosome 17, to tumorigenic BP1-E cells increased the apoptotic ratios and decreased the nuclear instability ratios, thus showing that the microsatellite instability and loss of heterozygosity induced by BP in these chromosomes of MCF-10F cells affect the control of cell death mechanisms. (C) 2003 Elsevier B.V. All rights reserved.

Isolation and characterization of microsatellite loci in Pseudoplatystoma corruscans (Siluriformes : Pimelodidae) and cross-species amplification

A total of five polymorphic microsatellites loci from Pseudoplatystoma corruscans were isolated and characterized. A population survey involving 43 specimens resolved a large number of alleles (range seven to eight among loci) and high observed heterozygosity (0.500-0.615), indicating its usefulness in population genetics studies. Cross-species amplification was successful in four other Pimelodidae species.

Genetic differentiation among ten populations of the genus Neoplecostomus (Teleostei: Loricariidae) from the upper Parana River basin

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)