OBJECTIVES: The phosphatidylinositol 3-kinase/AKT axis is an important cell-signaling pathway that mediates cell proliferation and survival, two biological processes that regulate malignant cell growth. The phosphatidylinositol 3-kinase CA gene encodes the p110α subunit of the phosphatidylinositol 3-kinase protein. There are phosphatidylinositol 3-kinase CA mutations in several types of human tumors, and they are frequently observed in breast cancer. However, these mutations have not been investigated in Brazilian breast cancer patients. METHODS: PCR-SSCP and direct DNA sequencing were performed to identify phosphatidylinositol 3-kinaseCA exon 9 and exon 20 mutations in 86 patients with sporadic breast cancer. The relationships between PIK3CA mutations and patient clinicopathological characteristics and survival were analyzed. The presence of the TP53 mutation was also examined. RESULTS: Twenty-three (27%) of the 86 primary breast tumors contained PIK3CA mutations. In exons 9 and 20, we identified the hotspot mutations E542K, E545K, and H1047R, and we identified two new missense mutations (I1022V and L1028S) and one nonsense (R992X) mutation. Phosphatidylinositol 3-kinase CA exon 20 mutations were associated with poor overall survival and TP53 gene mutations. CONCLUSIONS: Phosphatidylinositol 3-kinase CA mutations are common in tumors in Brazilian breast cancer patients, and phosphatidylinositol 3-kinase CA and TP53 mutations are not mutually exclusive. Phosphatidylinositol 3-kinase CA exon 20 mutations are associated with poor survival, and they may be useful biomarkers for identifying breast cancer patients with aggressive tumors and for predicting the response to treatment with PI3K pathway inhibitors.

CONTEXTO: Mutações nos genes ras têm sido associadas a diversos tumores sólidos humanos. Membros da família de genes ras (Ki-ras, Ha-ras e N-ras) são estruturalmente relacionados e codificam para uma proteína (p21) que desempenha papel importante na regulação da transdução de sinal e crescimento celular.OBJETIVOS: Estudar a ocorrência de mutações nos genes Ki-ras e Ha-ras. Também estudamos a expressão do gene Ha-ras em 32 dos tumores de cabeça e pescoço. TIPO DE ESTUDO: Série de casos.PARTICIPANTES: 60 tumores de cabeça e pescoço e 28 nasoangiofibromas obtidos através de cirurgia.LOCAL: Hospital A. C. Camargo e Hospital das Clínicas da Universidade de São Paulo, respectivamente.TESTE DIAGNÓSTICO: Pela técnica de PCR-SSCP examinamos a ocorrência de mutações nos genes Ki-ras e Ha-ras. O nível relativo do mRNA de Ha-ras em 32 dos tumores de cabeça e pescoço foi examinado por Northern blot. RESULTADOS: Nenhum dos tumores de cabeça e pescoço nem os nasoangiofibromas apresentaram evidência de mutação nos codons 12, 13, 59 e 61 dos genes Ki-ras e Ha-ras; contudo, 17 (53%) dos tumores, onde a expressão gênica pode ser examinada, demonstraram níveis aumentados do mRNA de Ha-ras quando comparados com o tecido normal do mesmo paciente.CONCLUSÕES: Nossos resultados demonstram, pela primeira vez, que mutações nos genes Ki-ras e Ha-ras não estão associadas ao desenvolvimento dos nasoangiofibromas e confirmam trabalhos anteriores que demonstram que mutações que ativam o gene ras estão ausentes ou raramente associadas aos tumores de cabeça e pescoço de pacientes ocidentais. Além disso, nossos resultados sugerem que o aumento de expressão de Ha-ras, e não a mutação desse gene, possa ser um fator importante no desenvolvimento e progressão dos tumores de cabeça e pescoço.

CONETXT: Ras gene mutations have been associated to a wide range of human solid tumors. Members of the ras gene family (Ki-ras, Ha-ras and N-ras) are structurally related and code for a protein (p21) known to play an important role in the regulation of normal signal transduction and cell growth. The frequency of ras mutations is different from one type of tumor to another, suggesting that point mutations might be carcinogen-specific. OBJECTIVES: To study the occurrence of Ki-ras and Ha-ras mutations. We also studied the relative level of Ha-ras mRNA in 32 of the head and neck tumors. DESIGN: Case series. SETTING: University referral unit. PARTICIPANTS: 60 head and neck tumors and in 28 Juvenile Nasopharyngeal Angiofibromas (JNA). DIAGNOSTIC TEST: Using PCR-SSCP we examined the occurrence of Ki-ras and Ha-ras mutations. The relative level of Ha-ras mRNA was examined by Northern blot analysis. RESULTS: None of the head and neck tumors or JNA samples showed evidence of mutations within codons 12, 13, 59 and 61 of Ki-ras or Ha-ras genes. However, 17 (53%) of the tumors where gene expression could be examined exhibited increased levels of Ha-ras mRNA compared with the normal tissue derived from the same patient. CONCLUSIONS: Our results demonstrate for the first time that mutations of Ki-ras and Ha-ras genes are not associated with the development of JNA and confirm previous reports indicating that activating ras mutations are absent or rarely involved in head and neck tumors from western world patients. Furthermore, our findings suggest that overexpression of Ha-ras, rather than mutations, might be an important factor in the development and progression of head and neck tumors.

The incidence of TP53 point mutations and loss of heterozygosity (LOH) of chromosome 17 in colorectal tumors was determined in a group of Brazilian patients. We screened DNA samples from tumors and distal normal mucosa of 39 patients with colorectal cancer, for TP53 mutations by PCR-SSCP (single-strand conformation polymorphism) analysis. Chromosome 17 LOH was investigated using six PCR-based polymorphic markers and one VNTR probe. TP53 mutations were demonstrated in 15/39 of the cases. Mutations were distributed among all exons examined (five to eight), the majority of them being G/C to A/T transitions. LOH of chromosome 17p and 17q was detected in 70 and 46% of the tumors, respectively. There was a significant association between TP53 mutations and LOH in chromosome 17p (P = 0.0035) and 17q (P = 0.03). Although no correlation was observed between TP53 genetic alterations and clinical/ pathological characteristics, the association of TP53 mutations with loss of both chromosome 17 arms may indicate that TP53 inactivation provokes an unstable phenotype in tumor cells in colorectal tumors.

Durante 2 anos consecutivos, foi realizado um experimento na Estação Experimental de Ribeirão Preto, SP, do Instituto Agronômico, em um Latossolo roxo de textura argilosa, cultivado com soja [Glycine max (L.) Merrill] cultivar IAC-11. O objetivo do trabalho foi estudar o efeito da inoculação de sementes e de doses de linuron (1,00 e 2,00 kg/ha) e de oryzalin (1,875 e 3,75 kg/ha), aplicados em pré-emergência, no crescimento da planta, na fixação simbiótica do nitrogênio e na produtividade de grãos de soja. Além dos tratamentos com herbicidas, foram mantidos dois controles, sendo um não inoculado e outro inoculado com a estirpe de Bradyrhizobium japonicum SMS-463(=29W). Os tratamentos com herbicidas foram todos inoculados. O delineamento experimental empregado foi em parcelas subdivididas com quatro repetições dispostas em blocos ao acaso. Foram realizadas amostragens aos 28, 42, 56, 70, 84 e 98 dias após a semeadura (DAS) no primeiro ano, e aos 28, 42, 56, 70, 84 e 105 DAS no segundo ano. Nos 2 anos avaliou-se o peso de matéria seca de raiz, parte aérea e nódulos, número de nódulos e produtividade de grãos; e no segundo, também a atividade da nitrogenase dos nódulos. De modo geral, não houve efeito benéfico da inoculação de sementes no crescimento e na nodulação de plantas de soja. Somente a atividade da nitrogenase foi beneficiada pela inoculação. Houve maior efeito na aplicação dos herbicidas no primeiro ano. O crescimento da planta foi favorecido pela aplicação de herbicidas. A atividade da nitrogenase foi reduzida pela aplicação dos herbicidas, sendo as doses maiores mais prejudiciais. Oryzalin afetou mais a atividade da nitrogenase que linuron. Apesar de ter havido efeito dos diferentes tratamentos nos parâmetros estudados estes não alteraram o rendimento de grãos.

Field investigation was conducted in 1986 and 1987 at Ribeirão Preto, SP, Brazil, to determine the effect of linuron (1.00 and 2.00 kg/ha) and oryzalin (1.875 and 3.75 kg/ha) on plant growth, nodulation, nitrogen fixation and yield of soybean [Glycine max (L.) Merril.] cv IAC - 11. The soil type used was a Red Latosol. Beside herbicide treatments there were two controls: one not inoculated and another inoculated with Bradyrhizobium japonicum strain SMS-463 (=29W). All herbicide treatments were inoculated. The experiment was arranged in a split plot randomized block design with four replicates. Samples were taken at 28, 42, 56, 70, 84 and 98 days after sowing (DAS) and herbicide application in the first year and at 28, 42, 56, 70, 84 and 105 DAS in the second year. In both years, root, shoot and nodule dry weight were evaluated, as well as grain productivity. In the second year, nitrogenase activity was also evaluated. Generally, there wasn't a positive response of seed inoculation on plant growth and nodulation; but nitrogenase activity was improved by seed inocutalion. In the first year of application, the use of herbicides enhanced plant growth but nitrogen fixation decreased, mainly with the highest rates of both herbicides. Nitrogenase activity was lower with both herbicides, and was more affected by oryzalin. No effect of treatments were observed on soybean grain yield.

No ano agrícola 1987/88, foi realizado um experimento de campo na Estação Experimental de Ribeirão Preto, do Instituto Agronômico, em latossolo roxo, textura argilosa, com a finalidade de pesquisar a ação dos herbicidas aplicados em pós-emergência no controle de plantas daninhas e sua persistência em solo, na cultura de amendoim (Arachis hypogaea L). O experimento foi em parcelas subdivididas com as parcelas distribuídas em blocos ao acaso com quatro repetições. Nas parcelas, estudaram-se os tratamentos referentes à presença e à ausência de inoculação, sendo o inoculante preparado com a mistura de estirpes de Bradyrhizobium sp. (SMS-319, SMS-400 e SMS-561); nas subparcelas, os herbicidas fomesafen (250g/ha), lactofen (192g/ha), fluazifop-p-butil (187g/ha), haioxifop-metil (240g/ha) e a mistura de fomesafen com fluazifop-p-butil (250 + 187g/ha), além de uma testemunha sem herbicida. Nas entrelinhas das subparcelas, tomaram-se ao acaso cinco pontos paraformar uma amostrado solo composta para análise de persistência dos produtos. As principais plantas daninhas presentes no experimento foram as gramíneas Cenchrus echinatus L. e Eleusine indica (L.) Gaerth, e as dicotiledôneas Alternanthera fícoidea(L.) R. Br. e Sidaspp. Aos 20 dias da aplicação dos herbicidas, as gramíneas foram 100% controladas pelos graminicidas fluazifop-p-butil e haioxifop-metil. Fomesafen e lactofen controlaram eficientemente A. fícoidea e, regularmente, Sida spp. A mistura de fomesafen com fluazifop-p-butil não apresentou vantagens em relação aos herbicidas isolados. Os resultados obtidos mostraram que não houve influenciada inoculação no controle de plantas daninhas e que os herbicidas não foram tóxicos às plantas de amendoim. Os herbicidas fluazifop-p-butil e haioxifop-metil, aos 28 dias, não mais causavam fitotoxicidade na planta-teste, mostrando uma persistência no solo inferior a esse período. Aos 28 dias, no tratamento não inoculado, fomesafen e lactofen ainda persistiam. Nenhum dos herbicidas persistiu a períodos superiores a 56 dias.

A field experiment was carried outon Dusky Red Latosol at the Ribeirão Preto Experiment Station, in 1987/88, to study the effect of post-emergence herbicides upon grasses and broadleaves weeds, their phytotoxicity and soil persistence upon peanut, Arachis hypogeea L, cultivar Tatu. At sowing time, part of the seeds were inoculated with a mixture of Bradyrhizobim sp. strains (SMS-319, SMS-400 and SMS-561). The seeds inoculated and an untreated check (factor A) were assigned at random to the each plots within each of four blocks. The following herbicides: fomesafen (250g/ha), lactofen (192g/ha), fluazifop-p-butyl (187g/ha) e haloxyfopmethyl (240g/ha) and fomesafen + fluazifop-p-butyl (250g/ha + 187g/ha) (factor B) were assigned at random within each plot. Composite the soil samples were taken on the subplots at random at three different periods to study the persistence of the herbicides. The effects of the herbicides upon the weeds were evaluated in the 10th and 20th day after the treatment (DAT); the soil persistence at 0, 28 and 56 DAT; the stand and grain yield at harvest. Seed inoculation did not affect the weed control. On the 20th day, grasses were fully controlled (100%) by fluazifop-p-butyl and haloxyfop-methyl. Fomesafen and lactofen were efficient to control Alternanthera ficoidea and regular to Sida spp. The mixture fomesafen + fluazifop-p-butyl did not present any advantage on the control of A. ficoidea, Sida spp., Cenchrus echinatus and Eleusine indica. Fluazifop-p-butyl and haloxyfop-methyl, at 28 DAT, did not cause phytotoxicity to the test-plant, showing a soil persistence lesser than 28 days. In the non-inoculated treatment, fomesafen and lactofen still persisted at 28 days. None of the herbicides used persisted for period over 56 days.