Analysis of CDKN1A polymorphisms: markers of cancer susceptibility?

The CDKN1A (TP21)(2) gene encodes a 21-kD protein that is a critical downstream mediator of wild-type TP53 and an important regulator of the cell cycle. Failure in the function of this gene would be expected to result in abnormal cell proliferation and transformation. Tumor-associated mutations of the coding region of the TP21 are rare. on the other hand, some TP21 polymorphisms have been identified and characterized by single base substitutions. In the present study, we investigated the potential role of TP21 gene polymorphisms in skin, head, and neck tumorigenesis. A total of 261 samples were examined by polymerase chain reaction single-strand conformational analysis, and one mutation at codon 31 and four polymorphisms in exons 2 (codon 55) and 3 [nucleotide (nt)590] and in promoter region (nt2298) were identified. In conclusion, this investigation confirmed the rarity of mutations in this gene, arguing against a role for TP21 mutations in skin, head, and neck cancers. Also, our results show significant differences in nt2298 allele frequencies between normal individuals and skin malignant tumors (P < 0.05). The results suggest that this polymorphism affects TP21 transactivator binding and may be important during the pathogenesis of skin cancer. (C) 2003 Elsevier B.V. All rights reserved.

Chromosome polymorphism and cytotype establishment in Bidens pilosa (Asteraceae)

Chromosome evolution, in the Bidens genus, was by polyploidy mechanisms. The basic number to the genus is X = 12 chromosomes, as all the Fitchinae subtribe. The association of chromosome number and environment in these plants was made some time ago, concerning the cytotypes of the species. In this paper, the chromosome number of nine Bidens pilosa populations was studied. The chromosome numbers found were 2n = 48, 70 and 72. In six of the nine populations analysed the most frequent number was 2n = 72. The Ibiuna population had 2n = 70 chromosomes and two had the lowest chromosome number (2n = 48). Observations of the B. pilosa population suggests that ploidy levels were related to a geographical gradient establishing the environmental altitudes. The data showed that lower ploidy levels, that is 2n = 48, were found at lower altitudes, while the populations higher ploidy levels were found in several environments, over a wide distribution.

Color polymorphism in Leptodactylus fuscus (Anura, Leptodactylidae): A defensive strategy against predators?

Color patterns are strongly related to defensive strategies in anurans. Some anurans present more than one morphotype. Leptodactylus fuscus, for example, present two morphotypes (with and without vertebral white line). The proportion of each pattern in nature is different, whereby there are always more individuals without stripes. Therefore, we speculated if this difference in the observed color pattern is due to unequal predation pressures (i.e. stronger over the striped morphotype), and/or if there is a genetic component related to autossomic heritage. To test the selective predation over the morphotypes, we prepared plasticine models of L. fuscus with both phenotypes and placed them in the field. We did not find evidence of predation selection and as we found significant relationships between the proportions of the phenotypes and Mendelian proportions, we suggest that the phenotypes observed in this species are genetically determined (involving dominant and recessive alleles) and may not have a defensive function.

DNA repair gene polymorphism is associated with the genetic basis of atherosclerotic coronary artery disease

Background: Atherosclerotic coronary artery disease (CAD) is a multifactorial process that appears to be caused by the interaction of environmental risk factors with multiple predisposing genes. It is nowadays accepted that increased levels of DNA damage induced by xenobiotics play an important role in the early phases of atherogenesis. Therefore, in this study, we focus on determining whether genetic variations in xenobiotic-metabolizing [glutathione-S-transferase theta 1 (GSTT1), glutathione-S-transferase mu 1 (GSTM1), cytochrome P450 IIEI (CYP2E1)] and DNA repair [X-ray cross-complementing group 1 (XRCC1)] genes might be associated with increased risk for CAD. Methods: A case-control study was conducted with 400 individuals who underwent subjected to coronary angiography. A total of 299 were patients diagnosed with effective coronary atherosclerosis (case group; >20% obstructive lesion), and 101 (control group) were individuals diagnosed as negative for CAD (<20% obstructive lesions). The polymorphism identifications for GSTM1 and GSTT1, and for CYP2E1 and XRCC1 genes were performed by polymerase chain reaction (PCR) amplification and by PCR-RFLP, respectively. Results and conclusions: The XRCC1 homozygous wild-type genotype Arg/Arg for codon 399 was statistically less pronounced in the case subjects (21.4%) than in controls (38.5%); individuals with the variant XRCC1 genotype had a 2.3-fold increased risk for coronary atherosclerosis than individuals with the wild-type genotype (OR=2.3, 95% CI=1.13-4.69). Conversely, no association between GSTM1, GSTT1, and CYP2E1gene polymorphisms and coronary atherosclerosis was detected. The results provide evidence of the role of DNA damage and repair in cardiovascular disease. © 2011 Elsevier Inc. All rights reserved.

Allelic variants of XRCC1 and XRCC3 repair genes and susceptibility of oral cancer in Brazilian patients

Background: The capacity for DNA repair is essential in maintaining cellular functions and homeostasis; however, this capacity can be altered based on DNA sequence variations in DNA repair genes, which may contribute to the onset of cancer. Many single-nucleotide polymorphisms (SNPs) in repair genes have been found to be associated with oral cancer. The aim of this study was to investigate the relationship between the presence of allelic variants Arg194Trp (rs:1799782) and Arg399Gln (rs: 25487) of XRCC1 gene and Thr241Met (rs: 861539) of XRCC3 gene and susceptibility to oral cancer. We also attempted to correlate the frequencies obtained for each of the SNPs to histopathological parameters. Methods: A case-control study was conducted with genomic DNA from 150 patients with oral squamous cell carcinomas and 150 controls. SNPs were genotyped by RFLP-PCR. Results: The presence of the polymorphic variants of the XRCC1 gene within codon 194 (OR 0.82, 95% CI: 0.44-1.51) and codon 399 (OR 0.94, 95% CI: 0.59-1.50) and within the XRCC3 gene (OR 0.72; 95% CI: 0.45-1.16) were not associated with an increased risk of oral cancer. A combinational analysis of SNPs in both genes indicated no association. The presence of the allelic variants of these two genes had no statistically significant effect on tumor differentiation, lymph node invasion or tumor size. Conclusions: These results suggest that allelic variants of XRCC1 and XRCC3 are not suitable markers for susceptibility to carcinomas of the oral cavity and are also not related to the later stages of such tumors. © 2012 John Wiley & Sons A/S.

Association among XRCC1, XRCC3, and BLHX gene polymorphisms and chromosome instability in lymphocytes from patients with endometriosis and ovarian cancer

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

Mannose-binding lectin gene polymorphism and risk factors for cardiovascular disease in postmenopausal women

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

Prevalence of the germline TP53 p.R337H mutation in families with multiple cases of cancer

Germline mutations in TP53 gene are associated with Li-Fraumeni syndrome (LFS) and its variants Li-Fraumeni-like (LFL). They predispose carriers to a wide variety of early onset tumors. In Brazil, there is a high frequency of a germline mutation in this gene (NC_000017.9: c.1010G>A; p.R337H) in Southern and Southeastern regions, due to a founder effect. It is estimated to be present in 0,3% ofthe local population, but only few families have been detected. Due to this significant divergence, the purpose of this study was to verify the effectiveness of wider criteria for detection of these individuals. Herein, clinical criteria were established, DNA samples were collected, analyzed by Restriction Fragment Length Polymorphism (RFLP) and sequenced. Thus, assessing the prevalence of this mutation in families with multiple cases of cancer. Based on our proposed criteria, one out of 31 patients (3,22%) was found to carry p.R337H mutation. The patient developed ductal invasive breast cancer at age 47, invasive adenocarcinoma of the lung at age 48 and soft-tissue sarcoma at age 49. In addition, an extensive cancer family history was referred, atypical for LFS, including a case of Ewing’s sarcoma. These outcomes indicate that the proposed criteria may detect probable carriers who did not fit previous LFS criteria. Nevertheless, additional studies, which might include a larger number of families and more stringent parameters, will be useful to improve screening sensibility

Fluorescent amplified fragment length polymorphism genotyping of human and animal Staphylococcus aureus isolates from dairy farms with manual milking

Fluorescence amplified fragment length polymorphism (fAFLP) was used to assess the genetic relatedness of 40 Staphylococcus aureus strains isolated from human and animal skin samples in seven dairy farms with manual milking. S. aureus was isolated from 11 out of 30 (36%) human skin samples and from 29 out of 100 (29%) teat skin samples from apparently healthy cows. Genomic DNA from each isolate was double-digested with EcoRI and MseI and complementary oligonucleotide adaptors were ligated to the restriction fragments. Pre-selective and selective, amplification reactions were performed, the amplified fragments were separated by electrophoresis in an ABI377 sequencer and analysed using GeneScan 3.1 and Genotyper 2.5. Three single isolates (a-c), a predominant cluster with 35 isolates (d) and another cluster with two isolates (e) were identified. Both clusters d and e included human and animal isolates genetically related, because the profiles had 90-100% homology. Since no cluster was comprised uniquely of human or animal isolates and given the close genetic relatedness among human and animal samples in the farms, the present findings support the. hypothesis that dairy workers can spread S. aureus through manual milking. (C) 2005 Elsevier B.V. All rights reserved.

Imunorreatividade da p53 associada à ausência de mutações no gene TP53 em linfomas caninos

Sabendo-se da influência das mutações no gene TP53 no desenvolvimento das neoplasias e da discrepância entre os resultados obtidos pelas técnicas de sequenciamento e imunoistoquímica, esta pesquisa teve como objetivo relacionar a sequência do TP53 com a imunorreatividade da p53. Foram obtidas amostras de linfoma de 12 cães. O diagnóstico histopatológico foi determinado pela classificação de Kiel. O imunofenótipo e a imunomarcação da p53 foram determinados por imunoistoquímica. Para reação com a p53, utilizou-se anticorpo policlonal anti-p53 (CM1) na diluição de 1:500. A região do gene TP53 compreendida entre os exons quatro e nove foi amplificada por PCR e submetida ao sequenciamento. Apesar dos resultados obtidos pela imunoistoquímica, nenhuma mutação foi encontrada nas sequências analisadas. Conclui-se que a imunorreatividade da p53 pela imunoistoquímica não pode ser atribuída à presença de mutações no domínio central do gene TP53.

Characterization and polymorphism screening of IGF-I and prolactin genes in Nelore heifers

Insulin growth factor I (IGF-I) and prolactin (PRL) are peptide hormones that exert complementary effects on reproductive traits by acting on folliculogenesis. In view of the lack of information about the IGF-I and PRL genes in Bos indicus, the objective of this study was to partially characterize the promoter regions of these genes and to screen animals of different ages at first pregnancy for the presence of polymorphisms in these regions. In addition, we determined whether polymorphisms influence the regulation of the two hormone genes, evaluating their association with sexual precocity.The animals were divided into three groups according to age at first pregnancy: 1) 100 heifers considered to be sexually precocious that became pregnant at 15-16 months of age, 2) 100 heifers that became pregnant during the normal breeding season at 24 months of age, and 3) 100 heifers that did not become pregnant until 24 months of age. For the IGF-I gene, PCR-RFLP-SnaBI analysis showed the presence of genotypes AB and BB at frequencies of 0.02 and 0.98, respectively. Sequencing of the IGF-I gene fragment revealed a single nitrogen base change from cytosine to thymine, corresponding to the restriction site of SnaBI. The polymorphisms identified in the 5'-flanking region of the IGF-I gene may serve as a basis for future studies of molecular markers in cattle. For the PRL gene, PCR-RFLP-HaeIII analysis showed the presence of only one migration pattern, a finding characterizing the region studied as monomorphic. The study of other regions in the IGF-I and PRL genes might provide molecular data that can be used in the future for the selection of sexually precocious animals.

Genetic diversity of natural populations of Anemopaegma arvense (Bignoniaceae) in the Cerrado of São Paulo State, Brazil

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

Estimativa de parâmetros genéticos da habilidade de permanência aos 48, 60 e 72 meses de idade em vacas da raça Caracu

Objetivou-se estimar a herdabilidade da característica habilidade de permanência (HP) em um rebanho de bovinos da raça Caracu visando sua utilização como critério de seleção. A característica em estudo foi definida como a probabilidade de a vaca estar presente no rebanho aos 48 (HP48), aos 60 (HP60) e aos 72 (HP72) meses, desde que possuíssem registros de pelo menos duas lactações nas específicas idades. Observações binárias, com zero (0 = fracasso) e um (1 = sucesso) foram designadas aos animais. Para análise da HP, foram utilizados dados de 5.487, 4.947 e 4.308 animais aos 48, 60 e 72 meses, respectivamente. Os componentes de variância e as herdabilidades foram estimados mediante inferência Bayesiana, via amostragem de Gibbs, pelo programa MTGSAM - threshold, utilizando-se um modelo touro. Foram utilizadas como variáveis explanatórias grupo de contemporâneos, classe de produção de leite na primeira lactação, classe de idade ao primeiro parto e sua interação. As análises forneceram estimativas médias de herdabilidade iguais a 0,28 ± 0,07 para HP48, 0,27 ± 0,07 para HP60 e 0,23 ± 0,07 para HP72. Os resultados evidenciaram que a característica HP apresentou variação aditiva em todas as idades estudadas e, portanto, pode ser empregada como critério de seleção para longevidade produtiva.

Polymorphism analysis of the hsp70 stress gene in Broiler chickens (Gallus gallus) of different breeds

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

FITOTOXICIDADE DE FUNGICIDAS, ACARICIDAS E INSETICIDAS, SOBRE O MAMOEIRO (Carica papaya L.) CULTIVAR SUNRISE SOLO IMPROVED LINE 72/12 em CONDIÇÕES DE CAMPO

A presente pesquisa teve como objetivo estudar os efeitos fitotóxicos de fungicidas, acaricidas e inseticidas e algumas associações entre eles, em plantas de mamoeiros (Carica papaya L.) cv. Sunrise Solo Improved Line 72/12, em condições de campo, no município de São Mateus -- ES, pertencente à maior região produtora do Estado. O experimento foi arranjado em delineamento de blocos casualizados, com 4 repetições e 03 plantas úteis por parcela. Foram utilizados os seguintes produtos, com as respectivas doses, para cada 100 L de água: chlorothalonil (Daconil PM-200g); mancozeb (Dithane PM -- 200g); oxicloreto de cobre (Reconil -- 400g); thiabendazole (Tecto 450 -- 100ml); dicofol + tetradifon (Carbax -- 200ml); triazophós (Hostathion 400 BR -- 150ml); óxido de fenbutatina (Torque 500 SC -- 60ml); e abamectin (Vertimec 18 CE -- 50ml): Analisou-se a fitotoxicidade dos produtos testados, em relação à altura da planta, nº de folhas, número de flores e frutos ; diâmetro do caule e queimaduras ou injúrias foliares. As datas das avaliações foram: 01 dia antes das pulverizações, 15 dias e 30 dias após as mesmas. Os fungicidas Daconil BR, Reconil e Tecto 450; o fungicida acaricida Dithane PM; os acaricidas Carbax e Torque 500 SC; e o inseticida-acaricida Vertimec 18 CE, aplicados isoladamente, não afetaram o crescimento e a produção das plantas, nem causaram injúrias nas folhas das mesmas. A associação de fungicidas e fungicida-acaricida, com os acaricidas, ou inseticida-acaricida, não mostrou nenhum efeito fitotóxico sobre os parâmetros de crescimento avaliados, nem causaram queimaduras ou injúrias foliares.