Chagas' disease is a parasitic disease with unsatisfactory treatment, mainly in chronic stage. This study aimed to evaluate the trypanocidal activity and action mechanisms of α/β-amyrin and its semi-synthetic derivatives, together with four isolated natural triterpenes, tested against trypomastigote and amastigote forms. The structure-activity relationship was suggested and cytotoxicity was measured. In general, greater polar compounds may have improved the selectivity to the protozoan. Action mechanisms were only performed for the amastigotes of Trypanosoma cruzi by evaluating the ultrastructural alterations, membrane permeability, mitochondrial membrane potential and cell volume, since the majority of compounds displayed promising antiamastigote activities. Triterpenes promoted changes on mitochondrial membrane potential and ultrastructural features that suggest autophagy processes. Both combinations between α/β-amyrin and 3-O-acetyl-11-oxo-α/β-amyrin and 3-O-acetyl-α/β-amyrin with benznidazole displayed synergistic effects against amastigotes and antagonistic effects on LLCMK2 cells. The antiamastigote activities, chemical derivatization, drug combinations and action mechanisms revealed to be crucial approaches toward this chronic disease.

Copaiba oil, an oleoresin extracted from Copaifera genus, has been widely used in popular medicine for the treatment of several diseases. The aim of this study was to investigate the antifungal activity of the copaiba oil and its isolated compounds caryophyllene oxide, copalic acid and acetoxycopalic acid against Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum gypseum strains, using microdilution method and microscopy techniques. It was found that the copaiba oil and the copalic acid were active against dermatophytes by minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) tests. The MIC and MFC of copaiba oil against T. rubrum, T. mentagrophytes and M. gypseum were 125 µg mL-1 (250 µg mL-1), 500 µg mL-1 (500 µg mL-1) and 250 µg mL-1 (250 µg mL-1), respectively. For copalic acid, the MIC and MFC were 50 µg mL-1 (100 µg mL-1), 100 µg mL-1 (100 µg mL-1) and 50 µg mL-1 (100 µg mL-1), respectively. Fluorescence microscopy and scanning electronic microscopy were used to investigate inhibition on hyphal growth by compounds, copaiba oil and copalic acid, showing a strong inhibition and an irregular growth pattern. Cell wall, cytoplasmic membrane and intracellular contents were also damaged. In conclusion, copaiba oil and copalic acid showed great activity against dermatophytes, being potential compounds for the development of antifungal drugs.

ABSTRACT Many infections worldwide are associated with bacterial biofilms. The effects of isolated neolignans (conocarpan and eupomathenoid-5) and the dichloromethane extract of Piper regnellii (Miq.) C. DC., Piperaceae, were tested against isolates of methicillin-resistant Staphylococcus aureus and methicillin-sensitive S. aureus biofilms and S. aureus planktonic cells. The dichloromethane extract presented better results than isolated neolignans against all of the biofilms tested, with a minimum inhibitory concentration <400 µg/ml for preformed biofilms and minimal biofilm inhibitory concentration of 15.6 µg/ml for biofilm formation. The minimum inhibitory concentration to planktonic cells was <12.5 µg/ml. These results indicate a good effect of the dichloromethane extract against methicillin-resistant S. aureus and methicillin-sensitive S. aureus biofilms and efficient prophylaxis.

Com o propósito de aumentar a atividade anticâncer demonstrada anteriormente pelas 1-fenilssubstituído-3-(2-tioxo-1,3,4-oxadiazol-5-il) β-carbolinas 1a-c, neste trabalho foram realizadas a síntese e a avaliação in vitro da atividade antitumoral de novas bases de Mannich 2-7(a-c), derivadas da introdução de diferentes grupos alquilamino(metil) na unidade 1,3,4-oxadiazol de 1a-c. Os derivados 1a-c e 2-7(a-c) foram também avaliados quanto às atividades antibacteriana e antifúngica. Adicionalmente, um estudo in silico das propriedades de ADME dos novos compostos sintetizados 2-7(a-c) foi realizado pela avaliação de seus parâmetros de Lipinski e de dados de área de superfície topológica polar (TPSA) e de porcentagem de absorção (% ABS).

With the purpose of activity enhancement of 1-substituted phenyl-3-(2-thioxo-1,3,4-oxadiazol-5-yl) β-carbolines 1a-c, reported as potential antitumor agents in our previous study, herein we report the synthesis and antitumor activity evaluation of several novel Mannich bases 2-7(a-c), by the introduction of different alkylamino(methyl) groups in the 1,3,4-oxadiazole unity of 1a-c. The antimicrobial activities of 1a-c and of 2-7(a-c) were also evaluated. Additionally, an in silico study of the ADME properties of novel synthesized β-carboline derivatives 2-7(a-c) was performed by evaluation of their Lipinski's parameters and topological polar surface area (TPSA) and percentage of absorption (% ABS) data.

Do extrato hidroetanólico das partes aéreas de Acanthospermum australe (Asteraceae) foram identificados uma lactona diterpênica, o acetato de acantoaustralida (1) e dois flavonoides: quercetina (2) e crisosplenol D (3). As estruturas foram identificadas através de técnicas espectroscópicas de RMN de ¹H e 13C, gHSQC, gHMBC, TOCSY, gNOESY, EM e pela comparação com dados da literatura.

From the hydroethanolic extract of the aerial parts of Acanthospermum australe (Asteraceae) a diterpene lactone, acanthoaustralide-1-O-Acetate (1) and two flavonoids: quercetin (2) and chrysosplenol D (3) were identified. The structures were determined though the use of spectroscopic techniques such as NMR (¹H, 13C{¹H}, gHSQC, gHMBC, TOCSY, gNOESY), MS and compared with the literature data.

A atividade biológica do extrato bruto e as várias frações obtidas de folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck foi avaliada em Leishmania amazonensis. Este estudo incluiu o processo de extração e o fracionamento biomonitorado pelo método de cromatografia de adsorção. Um aumento progressivo no efeito antileishmania foi observado durante o processo de purificação. O extrato hidroalcoólico solúvel em água (EBA) apresentou concentração inibitória 50% (IC50) igual a 167 µg/mL enquanto o extrato hidroalcoólico solúvel em acetato de etila (EBAcOEt) mostrou um IC50 de 30 mg/mL sobre o crescimento de formas promastigotas após 48 h de cultivo. A fração hexano (FHex) apresentou uma atividade antileishmania maior que o EBAcOEt com IC50 de 21,5 µg/mL. Análises de citotoxicidade indicaram que as concentrações tóxicas do EBA, EBAcOEt e das frações foram maiores para macrófagos J774G8 do que para os protozoários.

Biological activity of the crude extract and several fractions obtained from Piper regnellii var. pallescens was assessed on Leishmania amazonensis. This study included the extraction process and bioassay-guided fractionation by the adsorption chromatography method. A progressive increase in the antileishmanial effect was observed in the course of the purification process. The hydroalcoholic extract water soluble (EBA) had a 50% inhibitory concentration (IC50) at 167 µg/mL whereas the hydroalcoholic extract acetate soluble (EBAcOEt) showed an IC50 of 30 µg/mL against the growth of promastigote forms after 48 h of culturing. The hexan fraction (FHex) showed an antileishmanial activity greater than EBAcOEt with IC50 at 21.5 µg/mL. Analysis of cytotoxicity indicated that the toxic concentrations of the EBA, EBAcOEt, and fractions were higher for J774G8 macrophages than for the protozoans.

Descrições morfoanatômicas preliminares do caule e da folha de Acanthospermum australe (Loefl.) Kuntze foram realizadas a partir de material vegetal fresco e fixado. Secções transversais e paradérmicas, coradas com safranina/azul de astra e azul de toluidina foram analisadas ao microscópio. As superfícies do caule e da folha foram observadas através de microscopia eletrônica de varredura. A. australe apresenta, tanto no caule quanto na folha, tricomas tectores pluricelulares, unisseriados e de extremidades afiladas, e glandulares discóides, com pedúnculo curto unicelular, no qual se prendem cinco séries de células. A folha é anfiestomática e os estômatos são do tipo anomocítico, levemente proeminentes em relação ao nível das demais células epidérmicas. O caule apresenta crescimento secundário inicial, destacando-se os ductos secretores no parênquima cortical e medular.

Preliminary morphological and anatomical descriptions of stem and leaf of Acanthospermum australe (Loefl.) Kuntze were made in fresh and fixed material. Cross and superficial sections, stained with safranin/astra blue and Toluidine blue were analyzed at the light microscopic and the images were acquired by means the program Image Pro-Plus, version 4.0 (Media Cybertecnics). The surface of the stem and leaf were observed through scanning electronic microscopy. The species presents in the stem and leaf surface multicellular nonglandular trichome, uniseriate with a pointed edge and multicellular glandular trichome like a disk with a unicellular peduncle, in which is attached a series of five cells. The leaf presents anomocytic stomata in both surfaces a little prominent in relation to the level of the epidermal cells. The stem presents an initial secondary growth with evident secretory ducts in the cortex and pith.

Um novo triterpeno, 3beta,22ksi,23ksi-triidróxi-22ksi-acetóxi-24-metil-lanosta-8,24(28)-dieno-31-al, denominado pisosteral, e quatro triterpenos lanostanos conhecidos, 3beta,22ksi,23ksi-triidróxi-22ksi-acetóxi-24-metil-lanosta-8,24(28)-dieno (pisosterol), 3alfa,22ksi,23ksi-triidróxi-22ksi-acetóxi-24-metil-lanosta -8,24(28)-dieno (3-epi-pisosterol), 3beta,22ksi,23ksi-triidróxi-23ksi-acetóxi-24-metil-lanosta-8,24(28)-dieno e 3beta,22ksi,23ksi-triidróxi-22ksi-acetóxi-24-etil-lanosta-8,24(28)-dieno, foram isolados do corpo de frutificação do fungo Pisolithus tinctorius, coletado em uma plantação de Eucalyptus. De uma linhagem de P. tinctorius isolada de Pinus taeda, foram isolados quatro triterpenos conhecidos derivados do lanostano, 3beta,22ksi-diidróxi-24-metil-lanosta-8,24(28)-dieno, 3beta,22ksi-diidróxi-24-etil-lanosta-8,24(28)-dieno, 3beta-hidróxi-lanosta-8,24-dieno (lanosterol) and 3beta-hidróxi-lanosta-7(8),9,24-trieno (agnosterol). Essas substâncias foram identificadas a partir de métodos químicos e físicos, principalmente espectrometria de massas e ressonância magnética nuclear 1D e 2D.

A new triterpene, 3beta,22xi,23xi-trihydroxy-24-methyllanosta-8,24(28)-diene-31-al 22-acetate, named pisosteral, and four known lanostane triterpenes derivatives, 3beta,22xi,23xi-trihydroxy-24-methyllanosta-8,24(28)-diene 22-acetate (pisosterol), 3alpha,22xi,23xi-trihydroxy-24-methyllanosta-8,24(28)-diene 22-acetate (3-epi-pisosterol), 3beta,22xi,23xi-trihydroxy-24-methyllanosta-8,24(28)-diene 23-acetate and 3beta,22xi,23xi-trihydroxy-24-ethyllanosta-8,24(28)-diene 22-acetate, were isolated from fruiting bodies of Pisolithus tinctorius, collected in an Eucalyptus plantation. From cultivation of a strain of P. tinctorius isolated from Pinus taeda, four known triterpenes lanostane derivatives, 3beta,22xi-dihydroxy-24-methyllanosta-8,24(28)-diene, 3beta,22xi-dihydroxy-24-ethyllanosta-8,24(28)-diene, 3beta-hydroxylanosta-8,24-diene (lanosterol) and 3beta-hydroxylanosta-7(8),9,24-triene (agnosterol), were isolated. These compounds were identified on the basis of chemical and physical methods, mainly mass spectrometry and 1D and 2D nuclear magnetic resonance.

OBJETIVO: Avaliar a qualidade microbiológica de amostras de água tratada e não-tratada proveniente de comunidades urbanas e rurais e examinar a relação entre ocorrência de coliformes e a média de temperatura da água, e uma comparação dos níveis de precipitação de chuva. MÉTODOS: No período de 1996 a 1999, foram analisadas 3.073 amostras de água tratada (clorada) e não-tratada pelo método dos tubos múltiplos para determinar o número mais provável de coliformes totais e fecais. Destas, 1.594 provenientes de água de torneiras, 1.033 de reservatório para estocar água tratada, 96 de água de minas e 350 de poços particulares. Tais amostras foram obtidas na região de Maringá, Paraná, Brasil. RESULTADOS: O maior número de amostras contaminadas por TC (coliformes totais) (83%) e FC (coliformes fecais) (48%) foi observado em água não tratada. O índice de TC e FC foi maior nas amostras de reservatórios do que nas torneiras ao longo de sistema de distribuição. Entre as amostras de água tratada, foram encontradas bactérias do grupo coliforme em 171 dos 1.033 reservatórios amostrados. CONCLUSÕES: A observação de que mais de 17% da água potável tratada contêm coliformes sugere tratamento insuficiente ou recrescimento. Em água tratada, amostras positivas para TC e FC parecem ser similares e sazonalmente influenciadas. Dois diferentes períodos podem ser considerados para a ocorrência de amostras positivas para TC e FC: (i) período quente e úmido (Setembro-Março) com alta percentagem de amostras contaminadas; e (ii) período frio e úmido (Abril-Agosto) onde a positividade é baixa. Amostras positivas para TC e FC diminuem com o decréscimo da temperatura da água.

OBJECTIVE: To evaluate the microbiological quality of treated and untreated water samples came from urban and rural communities and to examine the relationship between coliforms occurrence and average water temperature, and a comparison of the rainfall levels. METHODS: A sample of 3,073 untreated and treated (chlorinated) water from taps (1,594), reservoir used to store treated water (1,033), spring water (96) and private well (350) collected for routine testing between 1996 and 1999 was analyzed by the multiple dilution tube methods used to detect the most probable number of total and fecal coliforms. These samples were obtained in the region of Maringá, state of Paraná, Brazil. RESULTS: The highest numbers water samples contaminated by TC (83%) and FC (48%) were found in the untreated water. TC and FC in samples taken from reservoirs used to store treated water was higher than that from taps midway along distribution lines. Among the treated water samples examined, coliform bacteria were found in 171 of the 1,033 sampling reservoirs. CONCLUSIONS: Insufficient treatment or regrowth is suggested by the observation that more than 17% of these treated potable water contained coliform. TC and FC positive samples appear to be similar and seasonally influenced in treated water. Two different periods must be considered for the occurrence of both TC and FC positive samples: (i) a warm-weather period (September-March) with high percentage of contaminated samples; and (ii) cold-weather period (April-August) were they are lower. Both TC and TF positive samples declined with the decreased of water temperature.

OBJETIVO: Avaliar a qualidade microbiológica de amostras de água tratada e não-tratada proveniente de comunidades urbanas e rurais e examinar a relação entre ocorrência de coliformes e a média de temperatura da água, e uma comparação dos níveis de precipitação de chuva. MÉTODOS: No período de 1996 a 1999, foram analisadas 3.073 amostras de água tratada (clorada) e não-tratada pelo método dos tubos múltiplos para determinar o número mais provável de coliformes totais e fecais. Destas, 1.594 provenientes de água de torneiras, 1.033 de reservatório para estocar água tratada, 96 de água de minas e 350 de poços particulares. Tais amostras foram obtidas na região de Maringá, Paraná, Brasil. RESULTADOS: O maior número de amostras contaminadas por TC (coliformes totais) (83%) e FC (coliformes fecais) (48%) foi observado em água não tratada. O índice de TC e FC foi maior nas amostras de reservatórios do que nas torneiras ao longo de sistema de distribuição. Entre as amostras de água tratada, foram encontradas bactérias do grupo coliforme em 171 dos 1.033 reservatórios amostrados. CONCLUSÕES: A observação de que mais de 17% da água potável tratada contêm coliformes sugere tratamento insuficiente ou recrescimento. Em água tratada, amostras positivas para TC e FC parecem ser similares e sazonalmente influenciadas. Dois diferentes períodos podem ser considerados para a ocorrência de amostras positivas para TC e FC: (i) período quente e úmido (Setembro-Março) com alta percentagem de amostras contaminadas; e (ii) período frio e úmido (Abril-Agosto) onde a positividade é baixa. Amostras positivas para TC e FC diminuem com o decréscimo da temperatura da água.

OBJECTIVE: To evaluate the microbiological quality of treated and untreated water samples came from urban and rural communities and to examine the relationship between coliforms occurrence and average water temperature, and a comparison of the rainfall levels. METHODS: A sample of 3,073 untreated and treated (chlorinated) water from taps (1,594), reservoir used to store treated water (1,033), spring water (96) and private well (350) collected for routine testing between 1996 and 1999 was analyzed by the multiple dilution tube methods used to detect the most probable number of total and fecal coliforms. These samples were obtained in the region of Maringá, state of Paraná, Brazil. RESULTS: The highest numbers water samples contaminated by TC (83%) and FC (48%) were found in the untreated water. TC and FC in samples taken from reservoirs used to store treated water was higher than that from taps midway along distribution lines. Among the treated water samples examined, coliform bacteria were found in 171 of the 1,033 sampling reservoirs. CONCLUSIONS: Insufficient treatment or regrowth is suggested by the observation that more than 17% of these treated potable water contained coliform. TC and FC positive samples appear to be similar and seasonally influenced in treated water. Two different periods must be considered for the occurrence of both TC and FC positive samples: (i) a warm-weather period (September-March) with high percentage of contaminated samples; and (ii) cold-weather period (April-August) were they are lower. Both TC and TF positive samples declined with the decreased of water temperature.