Inhibitory effect of pisosterol on human glioblastoma cell lines with C-MYC amplification.

Despite the remarkable progress in the characterization of the molecular pathogenesis of glioblastoma multiforme (GBM), these tumors remain incurable and, in most cases, refractory to aggressive cytotoxic treatments. We conducted a morphological and cytogenetic study in two GBM cell lines (U343 and AHOL1), before and after treatment with pisosterol (at 0.5, 1.0 and 1.8 µg ml⁻¹), a triterpene isolated from the fungus Pisolithus tinctorius. No significant alteration was observed in the morphology and frequency of chromosomal abnormalities in the cell lines analyzed after treatment with pisosterol. Using fluorescence in situ hybridization analysis with a locus-specific probe for C-MYC showed that 72% of U343 and 65% of AHOL1 cells contained more than two alleles of C-MYC before treatment. After treatment, no effects were detected at lower concentrations of pisosterol (0.5 and 1.0 µg ml⁻¹). However, at 1.8 µg ml⁻¹ of pisosterol, only 33% of U343 cells and 15% of AHOL1 cells presented more than two fluorescent signals, suggesting that pisosterol blocks the cells with gene amplification. Cells that do not show a high degree of C-MYC gene amplification have a less aggressive and invasive behavior and are easy targets for chemotherapy. Therefore, further studies are needed to examine the use of pisosterol in combination with conventional anti-cancer therapy.

Cocaine-induced status epilepticus and death generate oxidative stress in prefrontal cortex and striatum of mice.

Oxidative stress (OS) has been related to cocaine's actions and also to numerous nervous system pathologies, including seizures. The purpose of this work was to determine the alterations in glutathione (GSH) content, nitrite/nitrate and MDA levels after cocaine-induced toxicity. Male Swiss mice were injected (i.p.) with cocaine 90 mg/kg and observed during 1h. After this cocaine overdose some animals presented status epilepticus (SE) while some died after seizures. These animals were divided in two groups, SE and death. A group with an association of the antioxidant Vitamin E (Vit E, 400mg/kg, i.p.) plus Coc 90 (Vit E plus Coc 90) was undertaken to assess the neuroprotective effect of Vit E. Neurochemical analyses were carried out in prefrontal cortex (PFC) and striatum (ST). GSH levels increased only after cocaine-induced death in both areas studied. Cocaine-induced SE has increased nitrite/nitrate content in PFC and ST, while after death the increase was only in PFC. MDA (the lipid peroxidation marker) was elevated after SE and death in ST and only after death in PFC. Antioxidant treatment significantly reduced the GSH, nitrite/nitrate in ST and MDA levels. Only nitrite/nitrate content in PFC has not been decreased by Vit E pretreatment. The results relate that oxidative stress occurs after cocaine-induced toxicity mainly after death indicating that probably the increase of OS in the animal's brain leads to seizures and death, also showing a protective effect of Vit E in this process. Together with previous results this study contributes to the knowledge of cocaine-induced toxicity and possible in the near future to the use of antioxidants in the prevention of cocaine-induced CNS toxicity.

Larvicidal and nematicidal activities of the leaf essential oil of Croton regelianus.

The chemical composition of the leaf essential oil of Croton regelianus collected from wild plants growing in two different sites at Ceará State (Brazil) was analyzed by GC/MS and GC-FID. Twenty monoterpenoids, representing more than 96% of the chemical composition of the oils, were identified and quantified. The oils showed similar chemical composition but considerable variation in the levels of each constituent. Ascaridole (33.9-17.0%), p-cymene (22.3-21.6%), and camphor (13.0-3.1%) were the predominant constituents. The monoterpene ascaridole was isolated and characterized by spectroscopic data. The essential oils and the isolated compounds were tested against Aedes aegypti and Artemia sp. larvae, and the root knot nematode Meloidogyne incognita. The bioassay results show that the essential oil of C. regelianus and ascaridole were moderately active against the M. incognita, but strongly effective against both A. aegypti and Artemia sp. larvae.

Antitumor activity of pisosterol in mice bearing with S180 tumor.

It has been reported that pisosterol was active against cancer cells but lacked activity on the development of sea urchin eggs. Recently, it was reported that pisosterol induces differentiation in leukemia cell line. The present study evaluated the in vivo antitumor activity of pisosterol a triterpene isolated from Pisolithus tinctorius. Histopathological and morphological analyses of the tumor and the organs, including liver, spleen and kidney, were performed in order to evaluate toxicological aspects associated with pisosterol treatments. Antitumor activity of pisosterol (50 or 100 mg/m2) was confirmed in mice bearing Sarcoma 180 tumor cells. The tumor growth inhibition ratios were 43.0% and 38.7% for mice treated with pisosterol at 50 and 100 mg/m2, respectively. Histopathological analysis of liver, kidney and spleen was also evaluated. Liver and kidney were the major affected organs by pisosterol, although the observed alterations can be considered reversible.

Antibacterial activity of GUAVA, Psidium guajava Linnaeus, leaf extracts on diarrhea-causing enteric bacteria isolated from Seabob shrimp, Xiphopenaeus kroyeri (Heller) / Avaliação da atividade antibacteriana de extrato de folhas de goiabeira, Psidium guajava Linnaeus, sobre bactérias entéricas diarreiogênicas, isoladas de camarão sete-barbas, Xiphopenaeus kroyeri (Heller)

Guava leaf tea of Psidium guajava Linnaeus is commonly used as a medicine against gastroenteritis and child diarrhea by those who cannot afford or do not have access to antibiotics. This study screened the antimicrobial effect of essential oils and methanol, hexane, ethyl acetate extracts from guava leaves. The extracts were tested against diarrhea-causing bacteria: Staphylococcus aureus, Salmonella spp. and Escherichia coli. Strains that were screened included isolates from seabob shrimp, Xiphopenaeus kroyeri (Heller) and laboratory-type strains. Of the bacteria tested, Staphylococcus aureus strains were most inhibited by the extracts. The methanol extract showed greatest bacterial inhibition. No statistically significant differences were observed between the tested extract concentrations and their effect. The essential oil extract showed inhibitory activity against S. aureus and Salmonella spp. The strains isolated from the shrimp showed some resistance to commercially available antibiotics. These data support the use of guava leaf-made medicines in diarrhea cases where access to commercial antibiotics is restricted. In conclusion, guava leaf extracts and essential oil are very active against S. aureus, thus making up important potential sources of new antimicrobial compounds.

O chá de folhas de goiaba Psidium guajava Linnaeus é comumente usado como remédio nas gastrenterites e diarréias infantis por aqueles que não têm acesso a antibióticos. Esta pesquisa estudou o efeito antibacteriano sobre bactérias causadoras de diarréias, do óleo essencial e do extrato de folhas de goiabeira usando como diluente: metanol, hexano e acetato de etila. Os extratos foram testados sobre Staphylococcus aureus, Salmonella spp. e Escherichia coli. As bactérias testadas foram isoladas de camarão sete-barbas Xiphopenaeus kroyeri (Heller) usando-se como controle cepas padrão, de cada espécie. Das bactérias testadas, o melhor efeito inibitório foi observado sobre Staphylococcus aureus. O extrato de metanol apresentou maior inibição bacteriana. O óleo essencial mostrou ação inibitória contra S. aureus e Salmonella spp. Foi demonstrada alguma resistência das cepas isoladas de camarão aos antibióticos comerciais testados. Estes dados confirmam que remédios à base de folhas de goiabeira podem ser utilizados em casos de diarréia provocada por essas bactérias, quando o uso de antibiótico for restrito. Conclui-se que extratos de folhas de goiaba e o seu óleo essencial são muito ativos contra S. aureus, tornando-os importantes fontes em potencial de novos compostos antimicrobianos.

Nematicidal and larvicidal activities of the essential oils from aerial parts of Pectis oligocephala and Pectis apodocephala Baker

The chemical composition of the essential oils from aerial parts of Pectis apodocephala and Pectis oligocephala were analyzed by GC-MS. The essential oils of these species were predominantly constituted by monoterpenes. Geranial (42.9-44.5 percent), neral (32.2-34.2 percent) and alpha-pinene (10.7-11.4 percent) were the main constituents in the oil of P. apodocephala, while p-cymene (50.3-70.9 percent) and thymol (24.4-44.7 percent), were the prevalent compounds in the oil of P. oligocephala. The essential oils were tested against the root knot nematode Meloidogyne incognita and Aedes aegypti larvae survival. The results obtained show that both essential oils exhibited significant activity and could be considered as potent natural namaticidal and larvicidal agents.

A composição química dos óleos essenciais das partes aéreas de Pectis apodocephala e Pectis oligocephala foi analisada por CG-EM. Os óleos essenciais destas espécies foram predominantemente constituídos por monoterpenos. Geranial (42, 9-44, 5 por cento), neral (32, 2-34, 2 por cento) e alfa-pineno (10, 7-11, 4 por cento) foram os constituintes majoritários no óleo de P. apodocephala, enquanto p-cimeno (50, 3-70, 9 por cento) e timol (24, 4-44, 7 por cento), foram os compostos prevalentes no óleo de P. oligocephala. Ambos os óleos foram testados contra o nematóide Meloidogyne incognita e larvas do mosquito Aedes aegypti no terceiro estágio. Os resultados obtidos mostraram que os óleos exibem significante atividade e podem, portanto, ser considerados como potenciais agentes nematicida e larvicida naturais.

Pisosterol induces monocytic differentiation in HL-60 cells.

The aim of this study was to determine whether the antiproliferative effects observed for pisosterol, a cytotoxic triterpene isolated from Pisolithus tinctorius, are related to cell differentiation induction using HL-60 cell line as a model. Also, the effects of pisosterol on normal human cells were examined in peripheral blood mononuclear cells (PBMC). The effects on cell viability and morphological changes were the first indications showing that pisosterol induces HL-60 differentiation. The demonstration of blue tetrazolium reduction in HL-60 cells exposed to pisosterol demonstrated differentiation in a dose- and time-dependent manner, reaching a maximum effect after 72 h incubation at 5 microg/mL. Assays for alpha-naphthyl acetate esterase activity indicated that pisosterol triggers differentiation towards a monocytic cell-like pathway. The antiproliferative effect of pisosterol was determined by inhibition of DNA synthesis based on BrdU incorporation into HL-60 proliferating cells. It appears that pisosterol-treated cells, despite displaying a differentiated phenotype, continued to proliferate at all doses tested after 72 h, with a slightly decrease at 5 microg/mL. Apoptosis was observed in pisosterol-treated cells in a dose-dependent way. Nevertheless, after the same period of incubation, no cytotoxicity was detected in PBMC in the presence of pisosterol even at 25 microg/mL, providing some evidence that pisosterol may be selective for tumor cells. The mechanisms underlying the effect of pisosterol in leukemia cells indicates the induction of a monocytic cell-like differentiation, suggesting that this compound could be used in the development of new pharmacological tools with potential therapeutic value in the management of leukemia with fewer side effects.

Insecticidal activity of 2-tridecanone against the cowpea weevil Callosobruchus maculatus (Coleoptera: Bruchidae)

The effect of 2-tridecanone vapor on the cowpea weevil (Callosobruchus maculatus) development was determined. Seeds of cowpea were infested with adults and exposed to different doses of 2-tridecanone isolated from Pilocarpus microphyllus Stapf ex Holm, a plant species native from northeastern Brazil. The pure monoterpene was evaluated both undiluted as well as in the dilutions 1:10, 1:100 and 1:1,000 (v/v). The following parameters of the cowpea weevil life cycle were analyzed in response to decreasing doses of 2-tridecanone: number of eggs laid, percentage of egg hatching on seeds, percentage of adult emergence, adult weight at emergence, mean developmental time and number of adults emerged. Vapor of 2-tridecanone caused a significant (P < 0.05) reduction in the number of eggs laid, in the percentage of eggs hatched and in the number of emerged adults in infested seeds. The fumigant insecticidal effect of 2-tridecanone was mainly due to its ovicidal activity.

O efeito dos vapores da 2-tridecanona sobre o caruncho do feijão-de-corda (Callosobruchus maculatus) foi avaliado. Sementes de feijão-de-corda infestados com insetos adultos foram expostas a diferentes doses de 2-tridecanona isolada de Pilocarpus microphyllus, uma espécie nativa do Nordeste do Brasil. O monoterpeno puro foi utilizado nas diluições 1:10, 1:100 e 1:1000 (v/v). Os parâmetros da biologia do inseto foram analisados em função da resposta a doses decrescentes de 2-tridecanona: número de ovos postos por fêmea, percentagem de eclosão de ovos, percentagem de emergência de adultos, peso dos adultos recém-emergidos, tempo médio de desenvolvimento e número total de ovos emergidos. Diferenças significativas (P < 0.05) entre as doses de 2-tridecanona testadas foram observadas, para quatro dos seis parâmetros biológicos analisados. Os resultados obtidos indicaram que a 2-tridecanona é tóxica para C. maculatus, reduzindo significativamente (P < 0.05) o número de insetos emergidos após a infestação. Esse efeito foi causado principalmente pela significativa redução observada na eclosão dos ovos expostos ao vapor da substância.

Atividade larvicida de óleos essenciais contra Aedes aegypti L. (Diptera: Culicidae) / Larvicidal activity of essential oils against Aedes aegypti L. (Diptera: Culicidae)

Na busca por controle químico alternativo contra Aedes aegypti L., muitas pesquisas são desenvolvidas e estimuladas no intuito de se descobrirem novas substâncias inseticidas de origem vegetal. Neste trabalho, o efeito larvicida de dez óleos essenciais foi avaliado contra A. aegypti. Os óleos foram obtidos por arraste a vapor e sua composição química foi determinada por cromatografia gasosa acoplada a espectrometria de massa. Os óleos essenciais foram diluídos em solução aquosa de dimetil sufóxido nas concentrações: 100, 50, 10 e 1 mg/ml. A atividade larvicida, baseada na percentagem de larvas mortas, foi avaliada 24h após o tratamento. O óleo essencial de Vanillosmopsis arborea Baker induziu a maior atividade larvicida, com CL50 de 15,9 mg/ml e CL90 de 28,5 mg/ml., enquanto o de O. gratissimum L. apresentou a menor atividade com CL50 de 95,80 mg/ml e CL90 de 102,86 mg/ml. Os resultados indicam que os óleos essenciais avaliados, principalmente o de V. arborea, são compostos por substâncias com efeito larvicida contra A. aegypti.

In the search for alternative chemical control against Aedes aegypti L., many researches are developed and encouraged in order to find new insecticidal plant substances. In this work, the larvicidal effect of ten essential oils was tested on A. aegypti. The oils were extracted by steam distillation and their chemical composition determined by GL-chromatography coupled with massspectroscopy. The essential oils were diluted in aqueous solutions of dimethyl sulfoxide with concentrations of: 100, 50, 10, and 1 mg/ml. The larvicidal activity, based on the percentage oflarval mortality, was evaluated after 24h exposure to the treatments. The essential oil of Vanillosmopsis arborea Baker presented the highest larvicidal activity, with CL50 of 15.9 mg/ml and CL90 of 28.5 mg/ml. On the other hand the essential oil of O. gratissimum L. showed the lower activity with CL50de 95,80 mg/ml and CL90 de 102,86 mg/ml. The results indicate that the tested essential oils, and particularly the V. arborea, are composed by substances with larvicidal properties for A. aegypti.

Caracterizaçao química parcial e bioquímica de sementes de Bauhinia forficata link / Biochemical and chemical partial characterization of Bauhinia forficata link seeds

Sementes quiescentes de Bauhinia forficata foram submetidas à caracterizaçao bioquímica por medio de análise de ácidos graxos, (fraccionamiento de proteínas e atividade hemaglutinante específica. A análise elementar da semente mostrou grande quantidade de proteína total e de lipídeos com 21,24 por cento e 19.45 por cento, respectivamente. Na fraçao lipídica, o ácido linolêico foi o mais abundante com 46,47 por cento. Com exceçao das prolaminas, as diferentes françoes protéicas (albuminas, globulinas, glutelinas ácidas e básicas) apresentaram actividade hemaglutinante contra hemácias de coelho tratadas e nao tratadas com enzimas proteolíticas. A maior actividade hemaglutinante específica foi evidenciada na fraçao glutelinas ácidas (1.072,25 U.H./mgP) contra sangue de coelho tratado com tripsina. Os aminoácidos presentes em maior teor foram glutamina (16,20 por cento) e valina (11,07 por cento). Assim, por apresentarem alto valor energético as sementes de Bauhinia forficata sao uma possível fonte apcional na alimentaçao

Cytotoxic activity of pisosterol, a triterpene isolated from Pisolithus tinctorius (Mich.: Pers.) Coker & Couch, 1928.

Pisolithus tinctorius (Basidiomycete) is an ectomicorrhizal fungus found in the roots and soil surrounding of many species of eucalyptus and pine trees. The present work verified the cytotoxic potential of pisosterol, a triterpene isolated from P. tinctorius collected in the Northeast region of Brazil, on three different animal cell models: mouse erythrocytes, sea urchin embryos and tumor cells. Pisosterol lacked activity on mouse erythrocytes as well as on the development of sea urchin eggs, but strongly inhibited the growth of all seven tumor cell lines tested, especially the leukemia and melanoma cells (IC50 of 1.55, 1.84 and 1.65 microg/ ml for CEM, HL-60 and B16, respectively). The results found for pisosterol were compared with those of doxorubicin and etoposide.