In vitro antimalarial activity of fractions and constituents isolated from Tabebuia billbergii / Actividad antimalárica in vitro de fracciones y constituyentes aislados de Tabebuia billbergii

Introduction: in vitro antimalarial activity of naphthoquinones (1-5), isolated from Tabebuia billbergii (Bureau & K. Schum.) Standl., was investigated. Tabebuia billbergii, commonly known as guayacán, is a plant traditionally used in the Amazon in numerous conditions like bacterial and fungal infections, fever, syphilis, malaria, trypanosomiasis, as well as stomach and bladder disorders, and tumours. Objective: to study the dichloromethane extracts of both the trunk and the inner bark of Tabebuia billbergii and to demonstrate the antimalarial activity of some of its bioactive components. Methods: some bioactive components were evaluated for the antimalarial activity against Plasmodium berghei, by using the inhibition of the differentiation cycle of the parasite measure by the ³H-hypoxanthine incorporation and compared to that obtained for chloroquine. Results: conventional chromatographic techniques and bioassay-guided fractionation (Artemia salina) allowed isolating from the active fractions one naphthoquinone (lapachol) and four naphtho-furan-4,9-diones. These compounds proved to have an important antiplasmodial effect, with very encouraging IC50's, especially when compared to the results shown by Chloroquine in the same experiment. In addition, two triterpenes, β-sitosterol and stigmasterol, were obtained from the bark. Conclusions: the activity-guided fractionation (A. salina) of dichloromethane extracts of the trunk and the inner bark of Tabebuia billbergii led to the isolation and the identification of five quinonoid compounds with antiplasmodial effect. The significant inhibitory activity in vitro against Plasmodium berghei observed for compound 2-acetyl-naphtho-[2,3b]-furan-4,9-dione allow us to present them as a potential antimalarial compound.

Introducción: se evaluó la actividad antimalárica in vitro de una serie de naftoquinonas (1-5), aisladas de Tabebuia billbergii (Bureau & K. Schum.) Standl., que es conocida comúnmente como guayacán, una planta utilizada tradicionalmente en la Amazonía en numerosos problemas de salud como infecciones bacterianas y fúngicas, fiebre, sífilis, paludismo, tripanosomiasis, así como en problemas estomacales, tumores y trastornos de la vejiga. Objetivo: estudiar los extractos en diclorometano tanto del tronco como la corteza interna de Tabebuia billbergii y evaluar la actividad antimalárica de algunos de sus componentes bioactivos. Métodos: la actividad antimalárica contra Plasmodium berghei se evaluó en algunos componentes bioactivos, por la inhibición del ciclo de la diferenciación de la medida de los parásitos mediante la incorporación de 3H-hipoxantina y se comparó con la obtenida para la cloroquina. Resultados: a través de técnicas cromatográficas convencionales y el fraccionamiento guiado por bioensayo (Artemia salina) se aislaron de las fracciones activas, una naftoquinona (lapachol) y 4 nafto-furan-4,9-dionas. Estos compuestos presentaron un efecto antiplasmodial importante, con buenos valores de IC50, especialmente cuando se compara con los resultados mostrados por la cloroquina en el mismo experimento. Además, se obtuvieron de la corteza 2 triterpenos, β-sitosterol y estigmasterol. Conclusiones: el fraccionamiento guiado por Artemia salina de los extractos en diclorometano del tronco y la corteza interna de Tabebuia billbergii, condujo al aislamiento y la identificación de 5 compuestos de naturaleza quinoidal con efecto antiplasmódicol. La actividad in vitro contra Plasmodium berghei observada para el compuesto 2-acetil-nafto-[2,3 b]-furan-4,9-diona, permite proponerlo como un potencial compuesto antimalárico.

Decreased glycolytic metabolism accelerates apoptosis in response to 2-acetyl furanonaphthoquinone in K1735 melanoma irrespective of bcl-2 overexpression.

Solid tumors are often placed under stress conditions, such as glucose starvation which may result in topoisomerase II drug resistance. In this study, we investigated whether glucose deprivation or substitution by fructose regulates tumor cell apoptosis induced by 2-acetyl furanonaphthoquinone (FNQ). We now show that FNQ exerts much greater antitumor activity than either 7-methoxy 2-ethyl FNQ or 2-ethyl FNQ. Whereas 0.8 microM FNQ induces apoptosis after 16 hours in glucose-supplemented conditions irrespective of bcl-2 overexpression in K1735 melanoma, 0.5 microM FNQ is also effective within 12 hours in low glucose or in fructose-supplemented medium. Under the latter conditions, apoptosis-associated PARP cleavage and cytosolic cytochrome C are increased, together with induction and partial translocation to mitochondria of phosphorylated Jun-N-terminal kinase and massive upregulation of mitochondrial Mn superoxide dismutase. We propose that mitochondrial colocalization of these activities is important in this synergistic anti-tumor effect of FNQ and glucose depletion. Since glucose limitation slows proliferation and decreases efficacy of some genotoxic drugs that trigger apoptosis in rapidly dividing cells, we propose evaluating FNQ as a novel therapeutic anti-cancer adjuvant against slowly proliferating tumors.

Patterns of leaf epicuticular waxes in species of Clusia: taxonomical implications

El género Clusia L. (Clusiaceae) comprende unas 300 especies que ocurren desde México y el sur de EEUU hasta Bolivia y el sur de Brasil. Entre ellas se incluyen árboles y arbustos, hemiepifitas, epifitas y lianas. El análisis taxonómico del género se dificulta por la pobre preservación de las flores al ser secadas. Este trabajo explora la composición de ceras epicuticulares para caracterizar especies mediante marcadores químicos. Se analizó la composición del extracto obtenido de seis especies mediante lavado de la superficie foliar con hexano. Las especies pudieron separarse en base a la proporción de alcanos, >90% del total en Clusia rosea, C. orthoneura y C. minor, a la presencia de los triterpenos a-amirina y lupeol en C. multiflora, y de friedelina y taraxerol, conjuntamente con C33 y C35 en C. grandiflora y C. schomburgkiana. Los resultados sugieren que la proporción de alcanos y triterpenoides de ceras epicuticulares tiene importancia taxonómica y puede ser utilizada para separar especies o secciones infragenéricas.

The genus Clusia L. (Clusiaceae) encompasses ca. 300 species and occurs from southern USA and Mexico, to southern Brazil and Bolivia. It includes free-standing trees and shrubs, hemiepiphytes, epiphytes, and lianas. Taxonomic analysis of this genus is difficult because of the poor preservation of floral material after drying. This work explores the composition of epicuticular waxes in order to allow characterization, at the species level, using chemical markers. The six species analyzed could be separated using the relative quantity of hexane-soluble compounds extractable from the leaf surface, which amount to >90% in Clusia rosea, C. orthoneura, and C. minor, the presence of the triterpenes a-amyrin and lupeol in C. multiflora, and of friedelin and taraxerol, together with C33 and C35, in C. grandiflora and C. schomburgkiana. The results suggest that the relative proportions of alkanes and triterpenoids in epicuticular waxes may have taxonomic significance for separating species or infrageneric sections.

O gênero Clusia L. (Clusiaceae) compreende umas 300 espécies que ocorrem desde México e o sul dos EE.UU. até Bolívia e o sul do Brasil. Entre elas se incluem árvores e arbustos, hemiepífitas, epífitas e lianas. A análise taxonômica do gênero se dificulta pela pobre preservação das flores ao ser secadas. Este trabalho explora a composição de ceras epicuticulares para caracterizar espécies mediante marcadores químicos. Se analisou a composição do extrato obtido de seis espécies mediante lavado da superfície foliar com hexano. As espécies puderam separar-se em base à proporção de alcanos, >90% do total em Clusia rosea, C. orthoneura e C. minor, à presença dos triterpenos a-amirina e lupeol em C. multiflora, e de friedelina e taraxerol, conjuntamente com C33 e C35 em C. grandiflora e C. schomburgkiana. Os resultados sugerem que a proporção de alcanos e triterpenóides de ceras epicuticulares tem importância taxonômica e pode ser utilizada para separar espécies ou seções infragenéricas.

Relationship of Mcl-1 isoforms, ratio p21WAF1/cyclin A, and Jun kinase phosphorylation to apoptosis in human breast carcinomas.

Full length Mcl-1 is an anti-apoptotic protein consisting of two closely migrating 42/40kDa species. We now investigated the relationship of these isoforms to the expression of cell cycle stimulatory (cyclin A) and inhibitory (p21WAF1) proteins and to the induction of apoptosis in wt p53 MCF-7 and mutant p53 SKBR3 human breast carcinomas. The latter cells exhibited lower 42kDa Mcl-1, higher expression of cyclin A relative to that of p21WAF1, and apoptosis in response to okadaic acid, a phosphatase 1/2A inhibitor. The proteasome inhibitor MG-115 selectively increased expression of the 40kDa Mcl-1 isoform and induced p21WAF1, but also promoted preferential apoptosis in SKBR3 cells. Neither okadaic acid nor MG-115 caused comparable effects in MCF-7 cells. However, vanadate or acetyl furanonaphthoquinone induced the 40kDa Mcl-1 and greater Jun kinase (JNK) phosphorylation without apoptosis-associated PARP fragmentation in MCF-7 cells. Our data suggest that the higher susceptibility of SKBR3 cells to undergo apoptosis may be partly due to their greater proliferative potential (cyclin A), low expression of the anti-apoptotic 42kDa Mcl-1 isoform, and suboptimal JNK activation in response to stress.