Andiroba oil (Carapa guianensis Aubl) shows cytotoxicity but no mutagenicity in the ACPP02 gastric cancer cell line.

Andiroba (Carapa guianensis Aubl) is an Amazonian plant whose oil has been widely used in traditional medicine for various purposes, including anti-inflammation. Research reports indicate that the oil can confer antitumor activity due to the presence of fatty acids, which can directly influence cell death mechanisms. Thus, andiroba oil (AO) has gained interest for its potential to be used in antineoplastic therapies. Here, we report an in vitro analysis of the cytotoxic and mutagenic potential of AO in the gastric cancer cell line, ACP02. Cell survival was assessed by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, differential staining with ethidium bromide and acridine orange assessed apoptosis-necrosis, and mutagenesis was assessed by the micronucleus test. The apolar oil was first diluted in 0.1% dimethyl sulfoxide (DMSO) and then further diluted to six concentrations (0.01, 0.1, 1, 10 and 100 µg/mL and 1 mg/mL) in RPMI medium. Controls included RPMI alone (negative control) and 0.1% DMSO diluted in medium (vehicle control). The MTT test showed that AO significantly reduced cell viability (P < .05) only when the highest tested concentration was applied for 48 hours. The apoptosis/necrosis test showed that the highest concentration of AO induced cell death by apoptosis at 24 and 48 hours. There was no statistically significant increase in the frequency of micronuclei. The ability of the AO to decrease the viability of ACP02 cells via apoptosis, without exerting mutagenic effects, suggests that the oil could be useful as an alternative therapeutic agent for primary tumors of stomach cancer.

A novel function for kojic acid, a secondary metabolite from Aspergillus fungi, as antileishmanial agent.

Kojic acid (KA) is a fungal metabolite used as a topical treatment skin-whitening cosmetic agent for melasma in humans; however its potential as an anti-leishmanial agent is unknown. Chemotherapy is one of the most effective treatments for Leishmaniasis. However, the drugs available are expensive, invasive, require long-term treatment and have severe side effects. Thus, the development of new effective leishmanicidal agents is a necessity. In this study we investigated the anti-leishmanial effect of KA on L. amazonensis, following in vitro and in vivo infections. KA (50 µg/mL) was found to decrease the growth by 62% (IC50 34 µg/mL) and 79% (IC50 27.84 µg/mL) of promastigotes and amastigotes in vitro, respectively. Ultrastructural analysis of KA-treated amastigotes showed the presence of vesicles bodies into the flagellar pocket, and an intense intracellular vacuolization and swelling of the mitochondrion. During the in vitro interaction of parasites and the host cell, KA reverses the superoxide anions (O2-) inhibitory mechanism promoted by parasite. In addition, 4 weeks after KA-topical formulation treatment of infected animals, a healing process was observed with a high production of collagen fibers and a decrease in parasite burden. Thus, these results demonstrated the great potential of KA as an anti-leishmanial compound.

Kojic acid, a secondary metabolite from Aspergillus sp., acts as an inducer of macrophage activation.

KA (kojic acid) is a secondary metabolite isolated from Aspergillus fungi that has demonstrated skin whitening, antioxidant and antitumour properties among others. However, limited information is available regarding its effects on macrophages, the major cell involved in cell defence. The aim of the present study was to analyse whether KA affects functional properties related to macrophage activation, such as phagocytosis and spreading ability over a substrate. Treatment of resident macrophages with 50 µg/ml KA for 1 h induced both morphological and physiological alterations in cells. Immunofluorescence microscopy revealed enhanced cell spreading and an increase in cell surface exposure, associated with a rearrangement of microtubules, actin filaments and intermediate filaments. KA also potentiated phagocytosis by macrophages, as demonstrated by the increase in phagocytic activity towards yeast, when compared to untreated cells. KA increased the production of ROS (reactive oxygen species), but not NO (nitric oxide) production. Three tests were used to assess cell viability; MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide], NR (neutral red) uptake and PI (propidium iodide) exclusion test, which showed that macrophages maintain their viability following KA treatment. Results indicate that KA can modulate macrophage activation through cytoskeleton rearrangement, increase cell surface exposure, enhance the phagocytic process and ROS production. The study demonstrates a new role for KA as a macrophage activator.

Biotransformation of sucrose into 5-hydroxy-2-hydroxymethyl-γ-pirone by Aspergillus flavus

The sucrose hydrolysis and the preference of consumption of glucose instead of fructose were investigated for the production of 5-hydroxy-2-hydroxymethyl-γ-pyrone (HHMP) in the presence of Aspergillus flavus IOC 3974 cultivated in liquid Czapeck medium. Standardized 0.5g of pellets were transferred as inoculum into twelve conical flasks of 250 ml containing 100 ml of medium with different sucrose concentration, which was kept at 120 rpm and 28"C for 16 days without pH adjustment. Aliquots of 500μl of the broth culture were withdrawn at 24 h intervals and analyzed. The major yield of HHMP was 26g l-1 in 120g l-1 of sucrose. At these conditions, A. flavus produced an invertase capable of hydrolyzing 65 percent of total sucrose concentration in 24h, and an isomerase capable of converting fructose into glucose. In this work, it focused the preference for glucose and, then, of fructose by A. flavus and the strategy used to produce HHMP.

Foram investigadas a hidrólise da sacarose e a preferência pela glicose frente à frutose no processo de produção do 5-hidroxi-2-hidroximetil-γ-pirona (HHMP) na presença de Aspergillus flavus IOC 3974 cultivado em meio líquido Czapeck. Quantidades de 0,5g de pelletes foram utilizadas como inóculo. Doze frascos cônicos de 250 ml contendo 100 ml de meio de culturacom diferentes concentrações de sacarose foram utilizados.Os microrganismos foram cultivados a 120 rpm e 28"C por 16 dias sem ajuste do pH. O maior rendimento do HHMP foi 26g l-1em 120g l-1de sacarose. Nestas condições, A. flavus, foi capaz de produzir uma invertase possibilitando a hidrólise de 65 por cento da concentração total de sacarose em 24 horas, conjuntamente com a produção de uma isomerase que foi capaz de converter a frutose em glicose. Este trabalho está focalizado preferencialmente no consumo da glicose frente à frutose por A. flavus e na estratégia de produção do HHMP.

Avaliação de minerais em plantas medicinais amazônicas / Minerals evaluation in Amazonian medicinal plants

Amazônia brasileira oferece um apreciável potencial de plantas com propriedades terapêuticas, embora a maioria seja pouco conhecida. Dessa forma, com o objetivo de verificar a potencialidade nutricional de ervas medicinais, determinou-se a concentração de Ca, Mg, Fe, Cu e Zn nas folhas e nos chás das espécies: Piper callosum Ruiz & Pav., Piperaceae, Mikania lindleyana DC., Asteraceae e Arrabidaea chica (Humb. & Bonpl.) B. Verl., Bignoniaceae. As amostras de plantas depois de terem sido processadas, foram submetidas a digestão e em seguida realizada as leituras dos metais em um espectrofotômetro de absorção atômica. Para o chá de Arrabidaea chica foram detectados teores de Ca (6955 a 20058 mg/L), Mg (2390 a 3094 mg/L) e Fe (40 a 61 mg/L). Para o chá de Mikania lindleyana além da presença de altos valores de Ca (17722 a 22336 mg/L), Mg (4531 a 9370 mg/L) e Fe (20 a 87 mg/L) foram encontrados de 7 a 16 mg/L de Cu e 9 a 41 mg/L de Zn. O chá do Piper callosum apresentou em média 2036 a 4344 mg/L de Ca, 618 a 4023 mg/L de Mg e 39 a 60 mg/L de Fe. Comparando-se os resultados dos minerais com os valores recomendados pela Organização Mundial da Saúde, conclui-se que os metais presentes nos chás das plantas poderiam contribuir na complementação das dietas alimentares das pessoas que as utilizam.

The Amazonian Brazilian offers an appreciable potential of plants with therapeutic properties, although most are little known. In this way, with the objective of verifying the potentiality nutritional of medicinal herbs, a work was developed to determine the concentration of Ca, Mg, Fe, Cu and Zn in the leaves and in the teas of these species: Piper callosum Ruiz & Pav., Piperaceae, Mikania lindleyana DC., Asteraceae e Arrabidaea chica (Humb. & Bonpl.) B. Verl., Bignoniaceae. After the plants samples have been processed, they were submitted to digestion and soon afterwards the metals were analyzed in an spectrophotometer of Atomic Absorption. The results showed the follow yields: for the tea of Arrabidaea chica Ca were detected (6955 to 20058 mg/L), Mg (2390 to 3094 mg/L) and Fe (40 to 61 mg/L). For the tea of Mikania lindleyana besides the presence of high values of Ca (17722 to 22336 mg/L), Mg (4531 to 9370 mg/L) and Fe (20 to 87 mg/L) they were found from 7 to 16 mg/L of Cu and 9 to 41 mg/L of Zn. The tea of the Piper callosum presented 2036 to 4344 mg/L of Ca, 618 to 4023 mg/L of Mg and 39 to 60 mg/L of Fe. Being compared the results of the minerals with the values recommended by the Health World Organization, is possible that the present metals in the teas of the plants could contribute in the complementation of the people's alimentary diets that use these medicinal plants.

Antraquinonas e naftoquinonas do caule de um espécime de reflorestamento de Tectona grandi (Verbenaceae) / Anthraquinones and naphtoquinones from the bark of Tectona grandis (Verbenaceae) reforestation specimen

O fracionamento do extrato hexânico do caule de um espécime de reflorestamento de Tectona grandis (Verbenaceae), através de procedimentos fitoquímicos clássicos, levou ao isolamento das naftoquinonas lapachol e desidro-a-lapachona e das antraquinonas tectoquinona e obtusifolina. As estruturas das substâncias foram caracterizadas através da análise de métodos espectrométricos de RMN. Este é o primeiro estudo fitoquímico de um espécime de reflorestamento de Tectona grandis, no Brasil, sendo o objetivo principal deste trabalho a comprovação da presença de tectoquinona em espécimes cultivados.

The hexane extract of the bark of Tectona grandis (Verbenaceae) afforded two anthraquinones and two naphtoquinones. Their caracterizations were obtained through NMR spectroscopic techniques. This is the first phytochemical study of the bark of Tectona grandis reforestation specimen in Brazil. The main interest in this work is proving the presence of tectoquinone in reforestation specimen.