Avaliação da atividade antimalárica de extratos obtidos de algas marinhas no litoral do Rio Grande do Norte

Malaria is a major parasitic disease worldwide, accounting for about 500 million cases and causing 2 million to 3 million deaths annually. Four species are responsible for transmitting this disease to humans: Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae and Plasmodium ovale. The parasite resistance to antimalarial drugs and the usual limitations of the vector control implications are contributing to the spread of the disease. The most of significant advances in the search for new antimalarial drugs is based on natural components, the main ones being currently used antimalarial drugs derived from plants. Research on natural products of marine origin (particularly algae) show that some species possess antiplasmodial activity. Knowing that the coast of Rio Grande do Norte is home to several species of algae, the present study was to evaluate, for the first time, the antimalarial activity of ethanolic extracts of seaweed Spatoglossum schroederi, Gracilaria birdiae and Udotea flabellum against Plasmodium falciparum 3D7 strain tests and in vitro using the murine model (Plasmodium berghei) for evaluation in vivo. These species were ground, macerated with ethanol for 24 hours and the extracts concentrated in rotaevaporador (45 ° C ± 5 ° C). For in vitro tests, the extracts were diluted and tested at concentrations between 100 and 1.56 μg/ml (seven concentrations in triplicate), in order to obtain IC50 of each extract. The cytotoxicity tests with macrophages and BGM were performed using the MTT colorimetric assay. BGM macrophages and cells were distributed in 96 wells per plate (1x 105 to macrophages and 1x104 cells per well for BGM) and incubated for 24h at 37 ° C. The ethanol extracts were diluted and tested at concentrations of 100 to 1,56 μg/ml (seven concentrations in triplicate). After periods of 24 hours of incubation with the extracts, 100 μg of MTT was added to each well, and 3 hours elapsed, the supernatant was removed and added 200 μl of DMSO in each well. The absorbance of each well was obtained by reading on a spectrophotometer at 570 nm filter. To evaluate the acute toxicity in vivo, Swiss mice received a single dose (oral) 2000 mg/kg/animal of each extract tested. The parameters of acute toxicity were observed for 8 days. For in vivo tests, Swiss mice were inoculated with 1x105 erythrocytes infected with P. berghei. The treatment was given first to fourth day after infection with 0.2 ml of the extracts in doses of 1000 and 500 mg//g animal. The negative control group received 0.2 ml of 2% Tween-20, whereas the positive control group received sub-dose of chloroquine (5 mg/kg/animal). The assessment of antimalarial activity was done by suppressing suppressing the parasitemia at 5 and 7 days after infection. The growth inhibition of parasites was determined relative to negative control (% inhibition = parasitaemia in control - parasitemia in sample / parasitemia control x 100), the mortality of animals was monitored daily for 30 days The results showed that algae Spatoglossum schroederi and Udotea flabellum showed antimalarial activity in vitro, with reduced parasitemia of 70.54% and 54, respectively. The extracts of the three algae tested showed moderate to high cytotoxicity. Algae S. schroederi and U. flabellum were active against P. berghei only at doses of 500 mg / kg with reduction ranging from 54.58 to 52.65% for the fifth day and from 32.24 to 47.34% for the seventh day, respectively. No toxicity was observed in vivo at the dose tested, over the 8 days of observation. Although preliminary data, the bioactive components in those possible seaweed may be promising for the development of new anti-malarial drugs

Desenvolvimento de emulsões o/a contendo extrato glicólico de açai e avaliação da atividade fotoprotetora

Sunscreen use is the most common photoprotection alternative used by the population, and so these products should offer improved protection with broad - spectrum, UVA and UVB protection . Vegetal substances have recently been considered as resources for sunscreen formulations due to their UV spectrum absorption and antioxidant properties. The Euterpe oleracea Mart., popularly known as açai, in its che mical composition contain polyphenols compounds, such as anthocyanins and flavonoids , to which antioxidant properties have been attributed . The aim of this work was to develop O/W sunscreens emulsions con taining açai glycolic extract ( AGE) and to evaluate both their physical stability , safety and photoprotective efficacy. The safety of the extract was evaluated by in vitro phototoxicity and cytotoxicity tests. Emulsions containing AGE and sunscreens were formulated using different types and concentrations o f polymeric surfactant (Acrylates/C 10 - 30 Alkyl Acrylate Crosspolymer and Sodium Polyacrylate). The influence of two rheology modifiers (Polyacrylamide (and) C13 - 14/Isoparaffin (and) Laureth - 7 and Carbomer) on the stability was also investigated. Physical stability was evaluated by preliminary and accelerated studies. The macroscopic analyses, pH value and electrical conductivity determinations and rheological behavior were evaluated at different time intervals . The in vivo Sun Protect Factor ( SPF ) was determined according to the International Sun Protection Factor Test Method – 2006 and UVA Protection Factor (FPUVA), wavelength critical and reason SPF/FPUVA were performed according to the method Colipa 2011. The extract did not present cytotoxic ity and phototoxic ity . The stable emulsion containing 5% glycolic extract of açai and 1.0% of sodium poliyacrylate showed pseudoplastic and thixotropic behavior . The sunscreen emulsion containing açai glycolic extract showed a SPF 25.3 and PF - UVA = 14.97. Whe n açai glycolic extract was added in the emulsion sunscreen, no significant increase in the in vivo SPF and FPUVA values were observed. This emulsion showed 1.69 of the SPF/PF - UVA ratio and a critical wavelength value of 378 nm, so may therefore be conside red a sunscreen with UVA and UVB protection.

Avaliação dos efeitos tóxicos in vitro e in vivo do extrato hidroetanólico dos frutos de Genipa americana L. (Rubiaceae) em camundongos Swiss

The therapeutic use of medicinal plants has contributed since antiquity in a beneficial way for health. However, many species lacks of scientific evidence which provide basis for their use in therapeutic practice. In this context is the Genipa americana L. species (Rubiaceae), popularly known as jenipapo and used to treat syfilis, ulcer and hemorrhagic disturbs. It's also used against bruising, as tonic and as aphrodisiac. Due this species lacks toxicological studies, the aim of this study was to evaluate the toxicity in vivo (acute and sub-chronic toxicity) and in vitro (cytotoxicity) of the hydroethanolic extract from G. americana fruits. The hydroethanolic extract of G. americana fruits was prepared by maceration. A preliminary phytochemical analysis was performed to assess the presence of secondary metabolites in the extract. The cytotoxicity study of the extract (0.1, 1.0, 10, 100 and 1000 mg / 100 ul) were performed against normal cells (3T3) and tumor (786-0, HepG2 and B16), analyzed by the MTT assay. To evaluate the acute (single dose of 2000 mg / Kg) and subchronic (100, 500 and 1000 mg / kg for 30 days) toxicity Swiss mice of both sexes were used. At the end of the experiment, blood samples and organs were collected for analysis. Data between groups were compared by t test or ANOVA with Dunnett's post-test with 5% significance level. The phytochemical study of the extracts mainly indicated the presence of iridoids. Results for cytotoxicity tests showed up to 70% inhibition of B16 cell line at a dose of 1000 mg / 100 ul, and up to 29% inhibition of 786-0 at a dose of 10 ug / 100 ul. The extract did not cause death in 3T3 and HepG2 cells. During the in vivo assays, there were no animal deaths. Analysis of blood samples revealed that the animals submitted to the evaluation of acute toxicity had changes in AST and ALT, and that the animals evaluated for subchronic toxicity showed changes in the relative wet weight of the kidney and plasma urea concentration. No differences were observed between groups on histopathological evaluation of the collected organs. Despite the changes found in the in vivo toxicity tests, using the criteria described by the OECD Guidelines, it is suggested that the hydroethanolic extract of the fruits of the G. americana is classified as low toxicity. The cytotoxicity of the extract suggests that they have potential against melanoma cell lines (B16).

Avaliação dos efeitos tóxicos in vitro e in vivo do extrato hidroetanólico dos frutos de Genipa americana L. (Rubiaceae) em camundongos Swiss

The therapeutic use of medicinal plants has contributed since antiquity in a beneficial way for health. However, many species lacks of scientific evidence which provide basis for their use in therapeutic practice. In this context is the Genipa americana L. species (Rubiaceae), popularly known as jenipapo and used to treat syfilis, ulcer and hemorrhagic disturbs. It's also used against bruising, as tonic and as aphrodisiac. Due this species lacks toxicological studies, the aim of this study was to evaluate the toxicity in vivo (acute and sub-chronic toxicity) and in vitro (cytotoxicity) of the hydroethanolic extract from G. americana fruits. The hydroethanolic extract of G. americana fruits was prepared by maceration. A preliminary phytochemical analysis was performed to assess the presence of secondary metabolites in the extract. The cytotoxicity study of the extract (0.1, 1.0, 10, 100 and 1000 mg / 100 ul) were performed against normal cells (3T3) and tumor (786-0, HepG2 and B16), analyzed by the MTT assay. To evaluate the acute (single dose of 2000 mg / Kg) and subchronic (100, 500 and 1000 mg / kg for 30 days) toxicity Swiss mice of both sexes were used. At the end of the experiment, blood samples and organs were collected for analysis. Data between groups were compared by t test or ANOVA with Dunnett's post-test with 5% significance level. The phytochemical study of the extracts mainly indicated the presence of iridoids. Results for cytotoxicity tests showed up to 70% inhibition of B16 cell line at a dose of 1000 mg / 100 ul, and up to 29% inhibition of 786-0 at a dose of 10 ug / 100 ul. The extract did not cause death in 3T3 and HepG2 cells. During the in vivo assays, there were no animal deaths. Analysis of blood samples revealed that the animals submitted to the evaluation of acute toxicity had changes in AST and ALT, and that the animals evaluated for subchronic toxicity showed changes in the relative wet weight of the kidney and plasma urea concentration. No differences were observed between groups on histopathological evaluation of the collected organs. Despite the changes found in the in vivo toxicity tests, using the criteria described by the OECD Guidelines, it is suggested that the hydroethanolic extract of the fruits of the G. americana is classified as low toxicity. The cytotoxicity of the extract suggests that they have potential against melanoma cell lines (B16).

Avaliação do potencial genotóxico e citotóxico associado a queima artesanal da castanha de caju no Município de João Câmara

The Brazil is the third largest producer of cashew nuts in the world. Despite the social and economic importance of the cashew nut, its production is still carried out artisanally. One of the main problems encountered in the cashew production chain are the conditions under which the roasting of the nut occurs to obtain the kernel from the shell. In the present study was conducted a biomonitoring of the genotoxic and cytotoxicity effects associated with the elements from the cashew nut roasting in João Câmara - RN, semi-arid region of Brazil. To assess the genotoxic was used the bioassay of micronucleus (MN) in Tradescantia pallida. In addition, it was performed a comparative between the Tradescantia pallida and KU-20 and other biomarkers of DNA damage, such as the nucleoplasmic bridges (NBP) and nuclear fragments (NF) were quantified. The levels of particulate matter (PM1.0, PM2.5, PM10) and black carbon (BC) were also measured and the inorganic chemical composition of the PM2.5 collected was determined using X-ray fluorescence spectrometry analysis and the assessment of the cytotoxicity by MTT assay and exclusion method by trypan blue. . For this purpose, were chosen: the Amarelão community where the roasting occurs and the Santa Luzia farm an area without influence of this process. The mean value of PM2.5 (Jan 2124.2 μg/m3; May 1022.2 μg/m3; Sep 1291.9 μg/m3) and BC (Jan 363.6 μg/m3; May 70.0 μg/m3; Sep 69.4 μg/m3) as well as the concentration of the elements Al, Si, P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br and Pb obtained at Amarelão was significantly higher than at Santa Luzia farm. The genotoxicity tests with T. pallida indicated a significant increase in the number of MN, NBP and NF and it was found a negative correlation between the frequency of these biomarkers and the rainfall. The concentrations of 200 μg/mL and 400 μg/mL of PM2.5 were cytotoxic to MRC-5 cells. All together, the results indicated genotoxicity and citotoxicity for the community of Amarelão, and the high rates of PM2.5 considered a potential contributor to this effect, mainly by the high presence of transition metals, especially Fe, Ni, Cu, Cr and Zn, these elements have the potential to cause DNA damage. Other nuclear alterations, such as the NPBs and NFs may be used as effective biomarkers of DNA damage in tetrads of Tradescantia pallida. The results of this study enabled the identification of a serious occupational problem. Accordingly, preventative measures and better practices should be adopted to improve both the activity and the quality of life of the population. These measures are of fundamental importance for the sustainable development of this activity.

Produção e caracterização de quitooligossacarídeos produzidos pelo fungo Metarhizium anisopliae e avaliação da citotoxicidade em células tumorais

Chitosan is a natural polymer, biodegradable, nontoxic, high molecular weight derived from marine animals, insects and microorganisms. Oligomers of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) have interesting biological activities, including antitumor effects, antimicrobial activity, antioxidant and others. The alternative proposed by this work was to study the viability of producing chitooligosaccharides using a crude enzymes extract produced by the fungus Metarhizium anisopliae. Hydrolysis of chitosan was carried out at different times, from 10 to 60 minutes to produce chitooligosaccharides with detection and quantification performed by High Performace Liquid Chromatography (HPLC). The evaluation of cytotoxicity of chitosan oligomers was carried out in tumor cells (HepG2 and HeLa) and non-tumor (3T3). The cells were treated for 72 hours with the oligomers and cell viability investigated using the method of MTT. The production of chitosan oligomers was higher for 10 minutes of hydrolysis, with pentamers concentration of 0.15 mg/mL, but the hexamers, the molecules showing greater interest in biological properties, were observed only with 30 minutes of hydrolysis with a concentration of 0.004 mg/mL. A study to evaluate the biological activities of COS including cytotoxicity in tumor and normal cells and various tests in vitro antioxidant activity of pure chitosan oligomers and the mixture of oligomers produced by the crude enzyme was performed. Moreover, the compound with the highest cytotoxicity among the oligomers was pure glucosamine, with IC50 values of 0.30; 0.49; 0.44 mg/mL for HepG2 cells, HeLa and 3T3, respectively. Superoxide anion scavenging was the mainly antioxidant activity showed by the COS and oligomers. This activity was also depending on the oligomer composition in the chitosan hydrolysates. The oligomers produced by hydrolysis for 20 minutes was analyzed for the ability to inhibit tumor cells showing inhibition of proliferation only in HeLa cells, did not show any effect in HepG2 cells and fibroblast cells (3T3)

Atividade antimalárica de plantas medicinais da biorregião do Araripe-CE em modelo murino - Plasmodium berghei

Malaria, also popularly known as maleita , intermittent fever, paludism, impaludism, third fever or fourth fever, is an acute infectious febrile disease, which, in human beings, is caused by four species: Plasmodium falciparum, P. vivax, P. malariae and P. ovale. Malaria, one of the main infectious diseases in the world, is the most important parasitoses, with 250 million annual cases and more than 1 million deaths per year, mainly in children younger than live years of age. The prophylactic and therapeutic arsenal against malaria is quite restricted, since all the antimalarials currently in use have some limitation. Many plant species belonging to several families have been tested in vivo, using the murine experimental model Plasmodium berghei or in vitro against P. falciparum, and this search has been directed toward plants with antithermal, antimalarial or antiinflammatory properties used in popular Brazilian bolk medicine. Studies assessing the biological activity of medicinal plant essential oils have revealed activities of interest, such as insecticidal, spasmolytic and antiplasmodic action. It has also been scientifically established that around 60% of essential oils have antifungal properties and that 35% exhibit antibacterial properties. In our investigation, essential oils were obtained from the species Vanillosmopsis arborea, Lippia sidoides and Croton zethneri which are found in the bioregion of Araripe-Ceará. The chemical composition of these essential oils was partially characterized and the presence of monoterpenes and sesquiterpenes. The acute toxicity of these oils was assessed in healthy mice at different doses applied on a single day and on four consecutive days, and in vitro cytotoxicity in HeLa and Raw cell lines was determined at different concentrations. The in vivo tests obtained lethal dose values of 7,1 mg/Kg (doses administered on a single day) and 1,8 mg/Kg (doses administered over four days) for 50% of the animals. In the in vitro tests, the inhibitory concentration for 50% of cell growth in Hela cell lines was 588 μg/mL (essential oil from C. zethneri after 48 h), from 340-555 μg/mL (essential oil from L. sidoides, after 24 and 48 h). The essential oil from V. arborea showed no cytotoxicity and none of the essential oils were cytotoxic in Raw cell lines. These data suggest a moderate toxicity in the essential XVIII oils under study, a finding that does not impede their testing in in vivo antimalarial assays. Was shown the antimalarial activity of the essential oils in mice infected with P. berghei was assessed. The three species showed antimalarial activity from 36%-57% for the essential oil from the stem of V. arborea; from 32%-82% for the essential oil from the leaves of L. sidoides and from 40%-70% of reduction for the essential oil from the leaves of C. zethneri. This is the first study showing evidence of antimalarial activity with these species from northeast Brazil. Further studies to isolate the active ingredients of these oils are needed to determine if a single active ingredient accounts for the antimalarial activity or if a complex integration of all the compounds present occurs, a situation reflected in their biological activity

Avaliação da atividade antimalárica e citotóxica de plantas medicinais dos Biomas Caatinga e Amazônico

Resistance of Plasmodium falciparum to the usual antimalarials, as well as their adverse effects and high cost, has led to the search of new drugs against malaria. Several of these have been developed from medicinal plants based on ethnopharmacology, including the most widely used antimalarials today: quinine and artemisinin. In the present study schizonticide activity of extracts and fractions of a number of medicinal plants from the Caatinga and Amazon biomes were assessed based on ethnopharmacological and chemosystematic information. These included Ximenia americana, Maytenus rigida, Sideroxylon obtusifolium, Stryphnodendro coriaceum, Bowdichia virgiliodes, Schinopis brasiliensis and Picrolemma sprucei, the last, an Amazon species. Antimalarial tests of blood schizonticides were conducted in Swiss mice infected with P. berghei and in vitro against P. falciparum. In vitro cytotoxicity studies were carried out using HeLa, CHO, 3T3, Raw and HEPG2 cell lines. Except for X. americana, all species exhibited in vivo or in vitro antimalarial activity, inhibiting parasitic growth by up to 79%. Extracts exhibited moderate toxicity with dosedependent kinetics. In this sense, ethnopharmacological and chemosystematic approaches were shown to be useful and promising tools in the search of new drugs. These findings represent a significant contribution to scientific knowledge of the antimalarial potential of Brazilian flora, thereby opening perspectives for the development of new antimalarials