Alexandre de Barros Falcão Ferraz

Screening for the Presence of Hypericins in Southern Brazilian Species of Hypericum

Eight species of Hypericum (H. brasiliense, H. caprifoliatum, H. carinatum, H. connatum, H. cordatum, H. myrianthum, H. piriai and H. polyanthemum) growing in southern Brazil were analyzed by TLC and HPLC for the presence of hypericin and pseudohypericin. Although these polycyclic quinones have been identified in some Hypericum species, they were not detectable in the presently assessed samples. The chemotaxonomy of the taxon is briefly discussed.

Antigenotoxicity and Antioxidant Activity of Acerola Fruit (Malpighia glabra L.) at Two Stages of Ripeness

Genotoxic and antigenotoxic effects of acerola fruit at two stages of ripeness were investigated using mice blood cells. The results show that no ripeness stage of acerola extracts presented any genotoxic potential to damage DNA (Comet assay) or cytotoxicity (MTT assay). When antigenotoxic activity was analyzed, unripe fruit presented higher DNA protection than ripe fruit (red color) extract. The antioxidant capacity of substances also showed that unripe samples inhibit the free radical DPPH more significantly than the ripe ones. The results about determination of compounds made using HPLC showed that unripe acerola presents higher levels of vitamin C as compared to ripe acerola. Thus, vitamin C and the complex mixture of nutrients of Malpighia glabra L., and especially its ripeness stages, influenced the interaction of the fruit extract with the DNA. Acerola is usually consumed when ripe (red fruit), although it is the green fruit (unripe) that has higher potential as beneficial to DNA, protecting it against oxidative stress.

Protective effects of three extracts from Antarctic plants against ultraviolet radiation in several biological models

The photoprotective effect of the methanolic extracts of three Antarctic plant species - Deschampsia antarctica Desv., Colobanthus quitensis (Kunth) Bartl., and Polytrichum juniperinum Hedw. against UV-induced DNA damage was investigated in hamster lung fibroblasts (V79 cells) and in a biomonitor organism Helix aspersas, using comet assay. The protective, mutagenic, and antimutagenic profiles of these extracts were also evaluated using haploid strains of the simple eukaryote Saccharomyces cerevisiae, and antioxidant activity were investigated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay, as well as the hypoxanthine/xanthine oxidase assay. At the concentration range employed, the extracts were not cytotoxic or mutagenic to S. cerevisiae. In addition, the treatment with these extracts enhanced survival, and decreased induced reverse, frameshift, and forward mutations in a dose-response manner in all UVC doses employed. The plants extracts did not generate DNA strand breaks in V79 cells, and the treatment significantly decreased DNA damage induced by UVC. Extracts significantly decreased UVC-induced lipid peroxidation in V79 cells, showing a clear antioxidant property. Moreover, results of comet assay in V79 cells, employing Fpg, Endo III, and Endo V enzymes, demonstrated significant reduction of UVC-induced DNA damage after pre-incubation with these extracts. The treatment with all tested extracts were much less efficient against UVC-induced cytotoxicity in the yeast strain defective in photolyase as compared to the wild type strain, suggesting that this DNA repair pathway is stimulated by substances present in the extracts. All extracts showed a significant inhibitory effect in the hypoxanthine/xanthine oxidase assay, and they had the ability to scavenge DPPH. In H. aspersas, the treatment was able to protect against UVC-induced damage. In conclusion, D. antarctica, C. quitensis, and P. juniperinum extracts present photoprotective properties, which can be attributed to molecules, such as flavonoids and carotenoids, which act as UV-absorbing molecules and as antioxidants, as well as stimulate DNA-repair processes.

Rosmarinic acid as a protective agent against genotoxicity of ethanol in mice

The aim of the present work was to study the protective effects of rosmarinic acid against ethanol-induced DNA damage in mice. The antigenotoxic capacity of rosmarinic acid (100 mg/kg) was tested using pre-, co- and post-treatment with ethanol (5 g/kg). Peripheral blood (1 and 24 h) and brain cells (24 h) were evaluated using the comet assay and bone marrow was analyzed using the micronucleus assay (24 h). The results were compared to data of TBARS, enzymes with antioxidant activity, and DCFH-DA test. Peripheral blood and brain cells show that mean damage index (DI) and damage frequency (DF) values of ethanol with pre-treatment with rosmarinic acid group were significantly lower than in the ethanol group. In brain cells all different treatments with ethanol and rosmarinic acid showed significant decrease in DI and DF mean values when compared to ethanol group and negative control. No significant differences were observed in micronucleus frequency, activity of antioxidant enzymes and TBARS between groups. The DCFH-DA test show a reduction of 18% of fluorescence intensity when compare with ethanol group. The results show that rosmarinic acid could decrease the levels of DNA damage induced by ethanol, for both tissues and treatment periods.

Antigenotoxic effect of acute, subacute and chronic treatments with Amazonian camu-camu (Myrciaria dubia) juice on mice blood cells.

Myrciaria dubia, a plant native to the Amazon region, stands out as a fruit rich in vitamin C and other metabolites with nutritional potential. We evaluated the antioxidant, genotoxic and antigenotoxic potential of M. dubia juice on blood cells of mice after acute, subacute and chronic treatments. Flavonoids and vitamin C present in the fruit of M. dubia were quantified. In vitro antioxidant activity was evaluated by DPPH assay. Blood samples were collected for analysis after treatment, and the alkaline comet assay was used to analyze the genotoxic and antigenotoxic activity (ex vivo analysis using H(2)O(2)). The amount of vitamin C per 100mL of M. dubia was 52.5mg. DPPH assay showed an antioxidant potential of the fruit. No M. dubia concentration tested exerted any genotoxic effect on mice blood cells. In the ex vivo test, the juice demonstrated antigenotoxic effect, and acute treatment produced the most significant results. After the treatments, there was no evidence of toxicity or death. In conclusion, our data show that M. dubia juice has antigenotoxic and antioxidant activities, though with no genotoxicity for blood cells. Nevertheless, more in-depth studies should be conducted to assess the safety of this fruit for human consumption.

Corrective effects of acerola (Malpighia emarginata DC.) juice intake on biochemical and genotoxical parameters in mice fed on a high-fat diet

Acerola contains high levels of vitamin C and rutin and shows the corresponding antioxidant properties. Oxidative stress on the other hand is an important factor in the development of obesity. In this study, we investigated the biochemical and antigenotoxic effects of acerola juice in different stages of maturity (unripe, ripe and industrial) and its main pharmacologically active components vitamin C and rutin, when given as food supplements to obese mice. Initial HPLC analyses confirmed that all types of acerola juice contained high levels of vitamin C and rutin. DPPH tests quantified the antioxidant properties of these juices and revealed higher antioxidant potentials compared to pure vitamin C and rutin. In an animal test series, groups of male mice were fed on a standard (STA) or a cafeteria (CAF) diet for 13 weeks. The latter consisted of a variety of supermarket products, rich in sugar and fat. This CAF diet increased the feed efficiency, but also induced glucose intolerance and DNA damage, which was established by comet assays and micronucleus tests. Subsequently, CAF mice were given additional diet supplements (acerola juice, vitamin C or rutin) for one month and the effects on bone marrow, peripheral blood, liver, kidney, and brain were examined. The results indicated that food supplementation with ripe or industrial acerola juice led to a partial reversal of the diet-induced DNA damage in the blood, kidney, liver and bone marrow. For unripe acerola juice food supplementation, beneficial effects were observed in blood, kidney and bone marrow. Food supplementation with vitamin C led to decreased DNA damage in kidney and liver, whereas rutin supplementation led to decreased DNA damage in all tissue samples observed. These results suggest that acerola juice helps to reduce oxidative stress and may decrease genotoxicity under obesogenic conditions.

Effect of Agaricus blazei Murill on the Pulmonary Tissue of Animals with Streptozotocin-Induced Diabetes

The present study was designed to evaluate the oxidative stress as well as the therapeutic effect of Agaricus blazei Muril (A. Blazei) in rats with streptozotocin-induced diabetes. We used 25 Wistar rats, and DM was induced by injecting streptozotocin (70 mg/Kg i.p.). Agaricus blazei Muril was administered daily starting 40 days after disease onset. A. Blazei was tested as an aqueous extract for its phytochemical composition, and its antioxidant activity in vitro was also evaluated. Lipoperoxidation (LPO), and superoxide dismutase (SOD), catalase, and glutathione peroxidase activities were measured in the pulmonary tissue, as well as the presence of inducible nitric oxide synthase (iNOS), through immunohistochemistry. An anatomopathologic study was also performed. Phytochemical screening of A. Blazei detected the presence of alkaloids and saponins. The extract exhibited a significant antioxidant activity in the DPPH-scavenging and the hipoxanthine/xanthine oxidase assays. Pulmonary LPO increased in diabetic animals (0.43 ± 0.09; P < .001) as compared to the control group (0.18 ± 0.02), followed by a reduction in the A. Blazei-treated group (0.33 ± 0.04; P < .05). iNOS was found increased in the lung in diabetic rats and reduced in the A. Blazei-treated group. The pulmonary tissue in diabetic rats showed oxidative alterations related to the streptozotocin treatment. The A. Blazei treatment effectively reduced the oxidative stress and contributed to tissue recovery.

In vitro Cytotoxicity of Extracts from Brazilian Asteraceae

Aqueous and organic extracts of Asteraceae (Compositae) collected from the State of Rio Grande do Sul, Brazil, have been tested in vitro for cytotoxic activity against human solid tumour cell lines. Twenty-five species, 125 extracts in total, were screened against HT29 human colon adenocarcinoma cells and NCI-H460 human non-small cell lung cancer cells. Twenty-five extracts from 11 species demonstrated cytotoxicity at 100µg/ml against one or both of the cell lines tested. Further analysis was performed on the active extracts using three cell lines HT29, NCI-H460, and U373 human glioblastoma cells, to determine the IC 50 and the degree of tumour cell line selectivity. Extracts from Baccharis coridifolia, Baccharis ochracea, Eupatorium macrocephalum, Eupatorium pedunculosum and Stenachaenium riedelii all produced IC 50 values below 5µg/ml. Comparison of the IC 50 results between cell lines identified that Baccharis coridifolia, Baccharis ochracea, Eupatorium laevigatum and Pluchea sagittalis extracts produced differential sensitivity across the panel of three cell lines. These species are currently under further investigation with the ultimate objective of isolation and identification of the active principles responsible for the anti-proliferative activity.

Neurobehavioral and toxicological activities of two potentially CNS-acting medicinal plants of Piper genus

Plants from the genus Piper are economically useful and some species have been indicated because of their medicinal properties in the central nervous system. However, few studies about toxicity and neurobehavioral effects have been conducted. In this study, two Piper species, P. amalago and P. mikanianum were investigated in rats to determine acute toxicity and to evaluate the ansiogenic/ansiolytic properties in the elevated plus-maze and the effects on locomotion and exploration in an open field. Additionally, genotoxic activities were evaluated, using the comet assay in several tissues and the micronucleus assay in bone marrow. The phytochemical analysis of both Piper species leaves suggests the presence of amide, essential oils, flavonoids and phenolic compounds. The LD(50) of P. amalago and P. mikanianum were estimated as 2,545 and 1,661 mg/kg, respectively. The behavioral and genotoxic parameters were determined after an intraperitoneal administration of P. amalago (250 or 420 mg/kg) or P. mikanianum (160 or 270 mg/kg). Both plants decreased the number of entries and time spent in the open arms in the plus-maze test, indicating an anxiogenic effect. Only P. mikanianum affected locomotion and exploration in the open field behavior test. No genotoxic or mutagenic effect was observed. Our results suggest that these Piper species act on the central nervous system, without induce genetic toxicity.

Cytotoxic and leishmanicidal properties of garcinielliptone FC, a prenylated benzophenone from Platonia insignis.

Garcinielliptone FC (GFC), a natural prenylated benzophenone, was extracted from Platonia insignis Mart. (Clusiaceae), a native plant commonly known as bacuri and used in traditional Brazilian medicine for the treatment of skin diseases. The aim of this study was to evaluate the cytotoxic and leishmanicidal effects of GFC using in vitro models. The experimental data demonstrated that the polyisoprenylated benzophenone GFC possesses cytotoxic and leishmanicidal activities.