Antituberculosis Activities of Lapachol and ß-Lapachone in Combination with Other Drugs in Acidic pH.

Background: The treatment of multidrug-resistant tuberculosis (MDR-TB) is a challenge to be overcome. The increase of resistant isolates associated with serious side effects during therapy leads to the search for substances that have anti-TB activity, which make treatment less toxic, and also act in the macrophage acidic environment promoted by the infection. Objective: The aim of this study was to investigate lapachol and ß-lapachone activities in combination with other drugs against Mycobacterium tuberculosis at neutral and acidic pH and its cytotoxicity. Design: Inhibitory and bactericidal activities against M. tuberculosis and clinical isolates were determined. Drug combination and cytotoxicity assay were carried out using standard TB drugs and/or N-acetylcysteine (NAC). Results: Both naphthoquinones presented activity against MDR clinical isolates. The combinations with the first-line TB drugs demonstrated an additive effect and ß-lapachone+NAC were synergic against H37Rv. Lapachol activity at acidic pH and its association with NAC improved the selectivity index. Lapachol and ß-lapachone produced cell morphological changes in bacilli at pH 6.0 and 6.8, respectively. Conclusion: Lapachol revealed promising anti-TB activity, especially associated with NAC.

Larvicidal activity of piperovatine and dichloromethane extract from Piper corcovadensis roots against mosquitoes Aedes aegypti L / Actividad larvicida de piperovatina y extracto de diclorometano de las raíces de Piper corcovadensis contra mosquitos vectores del dengue Aedes aegypti L

The research of new substances capable of controlling the Aedes aegypti mosquito is urgent due to the increase in the transmission of the diseases such as dengue, chikungunya and Zika virus by the vector. Thus, the aim of this study was to evaluate the larvicidal activity of crude extract of Piper corcovadensis roots, a native plant from Brazil, and of the isolated compound piperovatine against larvae of A. aegypti by the larval immersion test. The lethal concentration that killed 50% (LC50) and 99% (LC99) of larvae was determined by Probit analysis. The results indicated high larvicidal activity on A. aegypti larvae for crude extract of Piper corcovadensis roots with LC50 of 4.86 µg/mL and LC99 of 15.50 µg/mL and piperovatine with LC50 of 17.78 µg/mL and LC99 of 48.55 µg/mL. This work opens new perspectives to the development of future products with crude extract of Piper corcovadensis roots and piperovatine that can be applied to mosquito control.

La investigación de nuevas sustancias capaces de controlar el mosquito Aedes aegypti es urgente debido al aumento en la transmisión de enfermedades como el dengue, el chikungunya y el virus Zika por el vector. Por lo tanto, el objetivo de este estudio fue evaluar la actividad larvicida del extracto crudo de las raíces de Piper corcovadensis, una planta nativa de Brasil, y del compuesto aislado piperovatine contra larvas de A. aegypti mediante la prueba de inmersión larvaria. La concentración letal que mató al 50% (LC50) y al 99% (LC99) de larvas se determinó mediante análisis Probit. Los resultados indicaron una alta actividad larvicida en larvas de A. aegypti para extracto crudo de las raíces de Piper corcovadensis con LC50 de 4.86 µg/mL y LC99 de 15.50 µg/mL y piperovatine con LC50 de 17.78 µg/mL y LC99 de 48.55 µg/mL. Este trabajo abre nuevas perspectivas para el desarrollo de futuros productos con extracto crudo de las raíces de Piper corcovadensis y piperovatine que pueden aplicarse al control de mosquitos.

Activity of Piperaceae extracts and fractions in the control of Phytomonas serpens / Atividade de extratos e frações de Piperaceae no controle de Phytomonas serpens

ABSTRACT: Protozoa of the genus Phytomonas are harmful parasites to several agricultural crops of economic importance. Due to their recognized biological activity, crude extracts of Piper aduncum, P. crassinervium, P. hispidum, and P. amalago leaves, were tested using the microdilution plate technique to assess the antiparasitic potential against Phytomonas serpens. Results showed that the ethanolic crude extract of P. crassinervium and P. amalago presented the best inhibitory concentration for 50% of the cells (IC50), 16.5 µg mL-1 in chloroform phase, and 18 µg mL-1 in aqueous phase, respectively, after 48 h treatment. Cytotoxicity analyses were performed using the colorimetric method of sulforhodamine-B in LLCMK2 mammalian cells. The chloroform phase of P. crassinervium was subjected to the fractionation process, in which the ethyl acetate and dichloromethane fractions obtained better IC50 values. Scanning electron microscopy (SEM) images showed alterations in the cell membrane of the treated parasites. The data obtained indicate a potential antiparasitic effect of the Piper species analyzed against P. serpens, being considered promising candidates for formulations of bioproducts to control the parasite.

RESUMO: Protozoários do gênero Phytomonas são parasitas prejudiciais a várias culturas agrícolas de importância econômica. Devido a sua atividade biológica reconhecida, extratos brutos de folhas de Piper aduncum, P. crassinervium, P. hispidum e P. amalago, foram testadas pela técnica de microdiluição em placa para avaliar o seu potencial antiparasitário contra Phytomonas serpens. Os resultados mostraram que o extrato bruto etanólico de P. crassinervium e P. amalago apresentaram as melhores concentrações inibitórias para 50% das células (IC50), 16,5 µg mL-1 na fase clorofórmio e 18 µg mL-1 na fase aquosa, respectivamente, após 48 h de tratamento. Análises de citotoxicidade foram realizadas através do método colorimétrico da sulforodamina-B, em células de mamíferos LLCMK2. A fase clorofórmio de P. crassinervium foi submetida ao processo de fracionamento, no qual as frações acetato de etila e diclorometano obtiveram melhores valores de IC50. Imagens de microscopia eletrônica de varredura (MEV) mostraram alterações na membrana celular dos parasitas tratados com fase aquosa de P. amalago. Os dados obtidos indicam potencial efeito antiparasitário das espécies de Piper analisadas contra P. serpens, sendo consideradas candidatas promissoras para formulações de bioprodutos para controle do parasito.

In vitro antioxidant potential and in vivo effects of Schinus terebinthifolia Raddi leaf extract in diabetic rats and determination of chemical composition by HPLC-ESI-MS/MS.

The present study investigated the in vitro and in vivo antioxidant potential and phytochemical composition of Schinus terebinthifolia, which is widely used in folk medicine for various therapeutic purposes. The in vitro analyses indicated that the hydroethanolic extract (HE) had 312.50 ± 0.50 mg GAE/g of total phenols. It also presented anti-DPPH• and anti-ABTS•+ activity, reduced phosphomolybden and metal ions and blocked the bleaching of ß-carotene. The HE at concentrations of 3.0 and 2.0 µg/mL had TRAP values of 2.223 ± 0.018 and 1.894 ± 0.026 µM Trolox, respectively. The HE increased the availability of antioxidants in plasma in treated animals in vivo. HPLC-ESI-MS/MS indicated the presence of 11 phenols: cumaric acid, (+)-catechin, myricetin-3-O-glicuronide, kaempferol-3-O-glucoside, myricetin, myricitrin, quercetin, gallic acid, methyl galate, pentagalloyl glucose and ethyl galate. Thus, S. terebinthifolia has potential for the prevention or treatment of diseases that are related to oxidative stress, such as diabetes mellitus.

Larvicidal potential of piperovatine in the control of cattle tick.

Rhipicephalus (Boophilus) microplus is one of the most important ectoparasites in cattle breeding worldwide, causing direct and indirect losses to animals and producers. Chemical acaricides are utilized in the control of cattle tick and the increase in the development of resistance by ectoparasites makes new alternative necessary. Therefore, research studies have been carried out using bioactive molecules that are quickly degraded and that reduce poisoning to appliers and non-target organisms, environmental contamination and development of resistance. Thus, this study aimed to isolate piperovatine from the roots of Piper corcovadensis, a native species to Brazil, and to evaluate the larvicidal activity against Rhipicephalus (Boophilus) microplus by larval packet test and in ex situ in an open environment. Piperovatine was isolated by classical column chromatography, and identified by 1H and 13C NMR. The lethal concentration (LC) of piperovatine that killed 50% (LC50) and 99% (LC99) of the larvae was determined by Probit analysis. The results indicated LC50 5.17 and LC99 25.41 µg/mL. LC99 was tested in ex situ in an open environment, and an efficiency of 96.63% was found, indicating that piperovatine kept the larvicidal action determined in in vitro test and in open environment. Therefore, this study shows new perspectives to develop products that can be applied in natural conditions to control this ectoparasite.

Anti-Mycobacterium tuberculosis activity of essential oil and 6,7-dehydroroyleanone isolated from leaves of Tetradenia riparia (Hochst.) Codd (Lamiaceae).

BACKGROUND: The global resurgence of tuberculosis (TB) and the development of drug resistance, as multidrug-resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis isolates, are a threat to TB control and have created a need for new and more effective anti-TB drugs. AIM: The current study evaluated the in vitro cytotoxicity and activity of Tetradenia riparia essential oil (TrEO) and 6,7-dehydroroyleanone pure compound against M. tuberculosis H37Rv and susceptible and resistant clinical isolates. METHODS: The in vitro activities of TrEO and 6,7-dehydroroyleanone were determined by Resazurin Microtiter Assay Plate (REMA). The cytotoxicity was evaluated in murine peritoneal macrophages by Alamar Blue assay. The cytotoxic effects were expressed as median concentration cytotoxicity (CC50) and the selectivity index (SI) was calculated. RESULTS: TrEO and 6,7-dehydroroyleanone showed activity against M. tuberculosis H37Rv with minimum inhibitory concentration (MIC) 62.5 µg/ml and 31.2 µg/ml, respectively. Both of them exhibited activities against resistant and susceptible M. tuberculosis clinical isolates with MIC values between 31.2 and 62.5 µg/ml. Cytotoxicity assays showed SI 1.9 and 7.9 for TrEO and 6,7-dehydroroyleanone, respectively. CONCLUSION: These results revealed that TrEO isolated from leaves of T. riparia and the pure compound 6,7-dehydroroyleanone display good activity against M. tuberculosis clinical isolates, including MDR isolates, with low cytotoxicity to murine macrophages. The 6,7-dehydroroyleanone compound is a potential candidate for anti-TB drug.

Relação entre a formação de radicais livres e a doença de Alzheimer: revisão sistemática / Relationship between free radicals formation and Alzheimer's disease: systematic review

Objetivo: realizar uma revisão sistemática sobre a relação entre os radicais livres e a doença de Alzheimer. Metodologia: a identificação dos artigos foi baseada nas diretrizes propostas pela declaração PRISMA e realizada em PubMed, Medline, Lilacs, IBECS e SciELO, com publicações entre 2006 e janeiro de 2016. Foram incluídos artigos que abordaram o acúmulo de radicais livres durante o envelhecimento e a relação entre o estresse oxidativo e a doença de Alzheimer. Resultados: 28 artigos, com resultados variados conforme os ensaios clínicos realizados e suas características e objetivos específicos, demonstrando envolvimento multissistêmico no estresse oxidativo. Conclusões: o estresse oxidativo celular que ocorre no envelhecimento é exacerbado na doença de Alzheimer devido ao acúmulo de peptídeo ß-amiloide, principalmente na região do hipocampo, provocando os sinais clínicos da doença.

Objective: to perform a systematic review of the relationship between free radicals and Alzheimer's disease. Methodology: the identification of articles was based on the guideline proposed by the PRISMA statement and done in PubMed, Medline, Lilacs, IBECS and SciELO with publications between 2006 and January 2016. Included articles addressed the accumulation of free radicals during aging and the relationship between oxidative stress and Alzheimer's disease. Results: 28 articles, with varying results according to clinical trials and their specific characteristics and objectives, demonstrating multisystemic involvement in oxidative stress. Conclusions: the cellular oxidative stress that occurs in aging is exacerbated in Alzheimer's disease due to the accumulation of ß-amyloid peptide, mainly in the hippocampus region, leading to clinical signs of the disease.

Cognitive capacity, style and quality of life of the elderly: a cross-sectional study / Capacidade cognitiva, estilo e qualidade de vida dos idosos: estudo transversal / Capacidad cognitiva, estilo y calidad de vida de los ancianos: estudio transversal

AIM: to verify the cognitive ability, style and quality of life of elderly primary care users. METHOD: a cross-sectional, correlational, analytical and descriptive study with a quantitative approach. A total of 252 semi-structured questionnaires were applied to the urban elderly, containing sociodemographic information, cognitive ability assessment and questions regarding style and quality of life. RESULT: there was a predominance of females with low schooling, presence of cognitive decline in 68.25% (n=172) of the elderly and a significant association between the Health Units (p=0,0351). DISCUSSION: it should be pointed out that elderly people who do not present a decline have a better score in the nutrition component, this data emphasizes that maintaining good eating habits positively influences cognitive ability. CONCLUSION: the interviewees are young elderly with cognitive decline. Emphasis is given to the importance of nurses acting as health promoters, favoring active aging.

OBJETIVO: verificar a capacidade cognitiva, estilo e qualidade de vida dos idosos usuários da atenção primária. MÉTODO: estudo transversal, correlacional, analítico e descritivo, com abordagem quantitativa. Foram aplicados 252 questionários semiestruturados aos idosos urbanos, contendo informações sociodemográficas, avaliação da capacidade cognitiva e questões referentes ao estilo e qualidade de vida. RESULTADO: houve predomínio do sexo feminino, com baixa escolaridade, presença de declínio cognitivo em 68,25% (n=172) dos idosos e associação significativa entre as Unidades de Saúde (p=0,0351). DISCUSSÃO: destaca-se que idosos que não apresentam declínio possuem melhor escore no componente nutrição, esse dado enfatiza que manter bons hábitos alimentares influencia positivamente na capacidade cognitiva. CONCLUSÃO: os entrevistados são idosos jovens que apresentam declínio cognitivo, enfatiza-se a importância de o enfermeiro atuar como promotor da saúde, favorecendo o envelhecimento ativo.

OBJETIVO: verificar la capacidad cognitiva, estilo y calidad de vida de los ancianos usuarios de la atención primaria. MÉTODO: estudio transversal, correlacional, analítico y descriptivo, con abordaje cuantitativo. Se aplicaron 252 cuestionarios semiestructurados a ancianos urbanos, conteniendo informaciones sociodemográficas, evaluación de la capacidad cognitiva y cuestiones referentes al estilo y calidad de vida. RESULTADO: predominó el sexo femenino, con baja escolaridad, presencia de decadencia cognitiva en 68,25% (n=172) de los ancianos y asociación significativa entre las Unidades de Salud (p=0,0351). DISCUSIÓN: se destaca que ancianos que no presentan decadencia tienen un mejor escore del componente nutrición, este dato enfatiza que mantener buenos hábitos alimentarios influye positivamente en la capacidad cognitiva. CONCLUSIÓN: los entrevistados son ancianos jóvenes que presentan decadencia cognitiva, se enfatiza la importancia de que el enfermero actúe como promotor de la salud, favoreciendo el envejecimiento activo.

Uso de plantas medicinais por idosos da atenção primária / Use of medicinal plants by primary care elderly

Este estudo tem como objetivo verificar o uso de plantas medicinais por idosos usuários da atenção primária. Caracterizou-se como um estudo transversal, correlacional, analítico e descritivo, com abordagem quantitativa. Realizado no oeste do Paraná, em 2016. Foram aplicados 252 questionários aos idosos, contendo informações sociodemográficas e questões relacionadas ao uso de plantas medicinais. Os resultados evidenciaram que 72,22% dos idosos utilizam plantas, entre as mais citadas destacam-se a Hortelã, Cidreira, Boldo e Alecrim. Referem utilizar por ser melhor para curar, gostam mais e não faz mal. Dado discordante quanto às indicações, sendo que 36,81% não indicam por medo de causar malefício. Quanto à toxicidade relataram efeitos adversos que não são apresentados por literatura científica. Ressalta-se a importância de uma orientação correta quanto à toxicidades de algumas plantas, bem como seus benefícios, e a importância do enfermeiro neste contexto já que atua como um promotor da saúde

Our study objective was to verify the use of medicinal plants by elderly users of the primary care. The study has a cross-sectional, correlational, analytical and descriptive design with a quantitative approach. It was carried out in the west of Paraná state, in 2016. We applied 252 questionnaires with socio-demographic information and questions about the use of medicinal plants. Our results showed that 72.22% of elderly use plants, and of the most cited ones, Peppermint,Lemon Grass, Boldo and Rosemary were noteworthy. Elderly report to use them because they are better for healing, they prefer them, and it does not harm. There was disagreement regarding the recommendations, where 36.81% does not recommend them for fear to cause harm. About toxicity, they reported adverse events that are not found in the scientific literature. We highlight the importance of a correct guidance regarding the toxicity of some plants, as well as their benefits, and the importance of the nurse in this context acting as a health promoter.

Evaluation of Chemical Composition and Antileishmanial and Antituberculosis Activities of Essential Oils of Piper Species.

Essential oils from fresh Piperaceae leaves were obtained by hydrodistillation and analyzed by gas chromatography mass spectrometry (GC-MS), and a total of 68 components were identified. Principal components analysis results showed a chemical variability between species, with sesquiterpene compounds predominating in the majority of species analyzed. The composition of the essential oil of Piper mosenii was described for the first time. The cytotoxicity of the essential oils was evaluated in peritoneal macrophages and the oils of P. rivinoides, P. arboretum, and P. aduncum exhibited the highest values, with cytotoxic concentration at 50% (CC50) > 200 µg/mL. Both P. diospyrifolium and P. aduncum displayed activity against Leishmania amazonensis, and were more selective for the parasite than for the macrophages, with a selectivity index (SI) of 2.35 and >5.52, respectively. These SI values were greater than the 1 for the standard drug pentamidine. The antileishmanial activity of the essential oils of P. diospyrifolium and P. aduncum was described for the first time. P. rivinoides, P. cernuum, and P. diospyrifolium displayed moderate activity against the Mycobacterium tuberculosis H37Rv bacillus, with a minimum inhibitory concentration (MIC) of 125 µg/mL. These results are relevant and suggests their potential for therapeutic purposes. Nevertheless, further studies are required to explain the exact mechanism of action of these essential oils.

[Level of awareness and the adequate application of sunscreen by beauticians]. / O conhecimento e a utilização de filtro solar por profissionais da beleza.

The scope of this research was to establish the level of awareness of beauticians regarding the importance of the application of sunscreen and to identify whether their patients had been properly instructed by these professionals. It involved a descriptive and exploratory study with interviews applying qualitative methodology among 30 beauticians. Data were gathered using the semi-structured interview technique in Maringá, in the southern state of Paraná. The data were analyzed using Atlas.ti software after applying quantitative analysis and response classification. Of those interviewed, 83.33% had a degree in Aesthetics, 20% attended ongoing training activities on sunscreen and 73.17% acquired sunscreen for its quality, though 86.67% were not familiar with sunscreens with natural anti-free radical components. Of those interviewed, 80% had never treated patients with skin cancer, though they reported having knowledge of care in relation to sun exposure and how to use the sunscreen and the relationship of these practices with the disease. The results showed that the recommendations and use of sunscreen by beauticians and users has been conducted in an adequate and conscientious manner.

O conhecimento e a utilização de filtro solar por profissionais da beleza / Level of awareness and the adequate application of sunscreen by beauticians

Resumo O objetivo desta pesquisa foi verificar o nível de instrução dos profissionais da área de estética em relação à utilização de filtro solar, e também identificar as orientações que estes passam aos seus pacientes. O estudo foi exploratório-descritivo, adotando a metodologia qualitativa ao entrevistar 30 profissionais da área. A coleta de dados foi feita por entrevistas semiestruturada na cidade de Maringá (PR). Os dados foram tratados com o software Atlas.ti a partir análise qualitativa e categorização das respostas. Dos entrevistados, 83,33% possuíam graduação em Estética, 20% participaram de atividade de formação continuada sobre o assunto, 73,17% adquirem o protetor solar pela qualidade, e 86,67% não conhecem filtros solares com componentes antirradicais livres naturais. Dos entrevistados, 80% nunca atenderam pacientes com câncer de pele, porém mencionaram ter conhecimento sobre os cuidados em relação à exposição solar e à forma de utilização do filtro solar e a relação dessas práticas com a doença. Os resultados mostraram que a indicação e o uso de filtro solar por profissionais da beleza vêm ocorrendo de maneira adequada e consciente.

Abstract The scope of this research was to establish the level of awareness of beauticians regarding the importance of the application of sunscreen and to identify whether their patients had been properly instructed by these professionals. It involved a descriptive and exploratory study with interviews applying qualitative methodology among 30 beauticians. Data were gathered using the semi-structured interview technique in Maringá, in the southern state of Paraná. The data were analyzed using Atlas.ti software after applying quantitative analysis and response classification. Of those interviewed, 83.33% had a degree in Aesthetics, 20% attended ongoing training activities on sunscreen and 73.17% acquired sunscreen for its quality, though 86.67% were not familiar with sunscreens with natural anti-free radical components. Of those interviewed, 80% had never treated patients with skin cancer, though they reported having knowledge of care in relation to sun exposure and how to use the sunscreen and the relationship of these practices with the disease. The results showed that the recommendations and use of sunscreen by beauticians and users has been conducted in an adequate and conscientious manner.

Traditional use of medicinal plants by elderly / Uso tradicional de plantas medicinais por idosos

Identificar o uso tradicional de plantas medicinais por idosos. Métodos: estudo exploratóriodescritivorealizado em Consórcio Intermunicipal de Saúde. Foram aplicados 351 questionários aos idosos,contendo informações sociodemográficas e questões relacionadas às plantas. Resultados: foi relatado o uso deplantas por 78,4% dos idosos, estas adquiridas em quintais. As mais citadas: hortelã, boldo, erva doce, capimlimão e camomila. Com relação ao motivo de uso, 33,3% afirmaram “não fazer mal à saúde”, 61,8% indicavamo uso para outras pessoas. A maioria dos idosos utiliza-se das plantas de forma segura, estando presentes nocotidiano destas pessoas como uma forma terapêutica. Conclusão: os idosos fazem uso das plantas medicinais,como um importante recurso terapêutico...

Antileishmanial activity of essential oil and 6,7-dehydroroyleanone isolated from Tetradenia riparia.

Tetradenia riparia plant is used as a traditional medicine in Africa for the treatment of inflammatory and infectious diseases as like parasitic. Therapy for leishmaniasis caused by Leishmania (Leishmania) amazonensis specie often fails, and the conventional drugs are toxic, expensive, require a long period of treatment, and adverse effects are common. The alternative therapies using natural products are inexpensive and have few or any adverse reaction. These reasons are sufficient to investigate the new natural therapeutic for leishmaniasis. We evaluated the potential of the essential oil (TrEO) and 6,7-dehydroroyleanone (TrROY) isolated from T. riparia on L. (L.) amazonensis promastigote and amastigote forms, cytotoxicity on human erythrocytes and murine macrophages, nitric production and inducible nitric oxide synthase (iNOS) mRNA expression. TrEO was the most effective to promote the Leishmania promastigote death. After 72 h incubation, the lethal dose of TrEO and TrROY that promoted 50% Leishmania death (LD50) were 0.8 µg/mL and 3 µg/mL, respectively. TrEO and TrROY were not cytotoxic to human erythrocytes, but TrROY was toxic to murine macrophages resulting in a low selectivity index. The transmission electronic microscopy showed that TrEO (0.03 µg/mL) was able to modify the promastigote ultrastructures suggesting autophagy as chromatin condensation, blebbing, membranous profiles and nuclear fragmentation. Infected-macrophages treated with TrEO (0.03 µg/mL) or TrROY (10 µg/mL) had an infection index decreased in 65 and 48%. TrEO did not induce iNOS mRNA expression or nitrite production in macrophages infected with Leishmania. TrROY and mainly TrEO promoted the Leishmania death, and TrROY showed loss toxicity to erythrocytes cells. Other compounds derived from T. riparia and the essential oil could be explored to develop a new alternative treatment for leishmaniasis.

Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae)

We evaluated the in vitro anti-Mycobacterium tuberculosis activity and the cytotoxicity of dichloromethane extract and pure compounds from the leaves of Calophyllum brasiliense. Purification of the dichloromethane extract yielded the pure compounds (-) mammea A/BB (1), (-) mammea B/BB (2) and amentoflavone (3). The compound structures were elucidated on the basis of spectroscopic and spectrometric data. The contents of bioactive compounds in the extracts were quantified using high performance liquid chromatography coupled to an ultraviolet detector. The anti-M. tuberculosis activity of the extracts and the pure compounds was evaluated using a resazurin microtitre assay plate. The cytotoxicity assay was performed in J774G.8 macrophages using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide colourimetric method. The quantification of the dichloromethane extract showed (1) and (2) at concentrations of 31.86 ± 2.6 and 8.24 ± 1.1 µg/mg of extract, respectively. The dichloromethane and aqueous extracts showed anti-M. tuberculosis H37Rv activity of 62.5 and 125 µg/mL, respectively. Coumarins (1) and (2) showed minimal inhibitory concentration ranges of 31.2 and 62.5 µg/mL against M. tuberculosis H37Rv and clinical isolates. Compound (3) showed no activity against M. tuberculosis H37Rv. The selectivity index ranged from 0.59-1.06. We report the activity of the extracts and coumarins from the leaves of C. brasiliense against M. tuberculosis.

Anti-Mycobacterium tuberculosis activity and cytotoxicity of Calophyllum brasiliense Cambess (Clusiaceae).

We evaluated the in vitro anti-Mycobacterium tuberculosis activity and the cytotoxicity of dichloromethane extract and pure compounds from the leaves of Calophyllum brasiliense. Purification of the dichloromethane extract yielded the pure compounds (-) mammea A/BB (1), (-) mammea B/BB (2) and amentoflavone (3). The compound structures were elucidated on the basis of spectroscopic and spectrometric data. The contents of bioactive compounds in the extracts were quantified using high performance liquid chromatography coupled to an ultraviolet detector. The anti-M. tuberculosis activity of the extracts and the pure compounds was evaluated using a resazurin microtitre assay plate. The cytotoxicity assay was performed in J774G.8 macrophages using the 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide colourimetric method. The quantification of the dichloromethane extract showed (1) and (2) at concentrations of 31.86 ± 2.6 and 8.24 ± 1.1 µg/mg of extract, respectively. The dichloromethane and aqueous extracts showed anti-M. tuberculosis H37Rv activity of 62.5 and 125 µg/mL, respectively. Coumarins (1) and (2) showed minimal inhibitory concentration ranges of 31.2 and 62.5 µg/mL against M. tuberculosis H37Rv and clinical isolates. Compound (3) showed no activity against M. tuberculosis H37Rv. The selectivity index ranged from 0.59-1.06. We report the activity of the extracts and coumarins from the leaves of C. brasiliense against M. tuberculosis.

New natural diterpene-type abietane from Tetradenia riparia essential oil with cytotoxic and antioxidant activities.

Tetradenia riparia (Hochstetter) Codd belongs to the Lamiaceae family and it was introduced in Brazil as an exotic ornamental plant. A previous study showed its antimicrobial, acaricidal and analgesic activities. Two compounds were isolated from essential oil of T. riparia leaves and identified as 9ß,13ß-epoxy-7-abietene (1), a new one, and 6,7-dehydroroyleanone (2), already reported for another plant. The structure of these compounds was determined by spectroscopic analysis and by comparison with literature data. The cytotoxic activities of the essential oil and compounds 1 and 2 were determined by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, and by tumor cells MDA-MB-435 (human breast carcinoma), HCT-8 (human colon), SF-295 (human nervous system) and HL-60 (human promyelocytic leukemia). The essential oil and compound 1 showed high cytotoxic potential of the cell lines SF-295 (78.06% and 94.80%, respectively), HCT-8 (85.00% and 86.54%, respectively) and MDA-MB-435 (59.48% and 45.43%, respectively). Compound 2 had no cytotoxic activity. The antioxidant activity was determined by 2,2-diphenyl-1-picryl-hydrazyl (DPPH), ß-carotene-linoleic acid system and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. The inhibitory concentration (IC50 in µg mL-1) for essential oil and compound 2 was, respectively 15.63 and 0.01 for DPPH; 130.1 and 109.6 for ß-carotene-linoleic acid and 1524 and 1024 for ABTS. Compound 1 had no antioxidant activity. By fractioning the oil, it was possible to identify two unpublished compounds: 1 with high cytotoxic potential and 2 with high antioxidant potential.

Evaluation of antifungal activity of extracts of Piper regnellii obtained by supercritical fluid extraction.

The antifungal activity of extracts obtained from Piper regnellii with supercritical carbon dioxide was tested against yeast and filamentous fungi. The most active extract was obtained from leaves extracts of P. regnellii at 40°C and 25 MPa, featuring a minimal inhibitory concentration of 3.9 µg/mL against Trichophyton mentagrophytes. Neolignans eupomatenoid-3, eupomatenoid-5, eupomatenoid-6 and conocarpan were present in all extracts. The results indicate the possibility of further studies on the use of extracts of P. regnellii obtained by supercritical extraction, as potential sources of bioactive compounds for use in medicine.

Antioxidant capacity of the leaf extract obtained from Arrabidaea chica cultivated in Southern Brazil.

Arrabidaea chica leaf extract has been used by people as an anti-inflammatory and astringent agent as well as a remedy for intestinal colic, diarrhea, leucorrhea, anemia, and leukemia. A. chica is known to be a good producer of phenolics. Therefore, in the present study, we investigated its antioxidant activity. The phenolic composition of A. chica leaves was studied by liquid chromatography coupled to diode array detection (LC-DAD) and liquid chromatography coupled to electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS), and isoscutellarein, 6-hydroxyluteolin, hispidulin, scutellarein, luteolin, and apigenin were identified. The extract from leaves of A. chica was tested for antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, ß-carotene bleaching test, and total reactive antioxidant potential (TRAP) method. The crude extract quenched DPPH free radicals in a dose-dependent manner, and the IC50 of the extract was 13.51 µg/mL. The ß-carotene bleaching test showed that the addition of the A. chica extract in different concentrations (200 and 500 µg/mL) prevented the bleaching of ß-carotene at different degrees (51.2% ±3.38% and 94% ±4.61%, respectively). The TRAP test showed dose-dependent correlation between the increasing concentrations of A. chica extract (0.1, 0.5, and 1.0 µg/mL) and the TRAP values obtained by trolox (hydro-soluble vitamin E) 0.4738±0.0466, 1.981±0.1603, and 6.877±1.445 µM, respectively. The 2 main flavonoids, scutellarein and apigenin, were separated, and their antioxidant activity was found to be the same as that of the plant extract. These 2 flavonoids were quantified in the plant extract by using a validated HPLC-UV method. The results of these tests showed that the extract of A. chica had a significant antioxidant activity, which could be attributed to the presence of the mixture of flavonoids in the plant extract, with the main contribution of scutellarein and apigenin.

Antileishmanial activity of amides from Piper amalago and synthetic analogs

Two natural amides isolated from the chloroform extract of Piper amalago L., Piperaceae, leaves, a hydrogenated derivative and seven synthetic analogs were tested against the promastigote and intracellular amastigote forms of Leishmania amazonensis. The antileishmanial activity was evaluated in terms of growth inhibitory concentration for 50% of protozoa (IC50). The cytotoxicity toward the J774A1 macrophages was evaluated in terms of the cytotoxic concentrations for 50% of macrophages (CC50). The ability to induce nitric oxide production was also investigated for all compounds. The saturated amide 7-(1,3-benzodioxol-5-yl)-1-(1-pyrrolidinyl)-1-heptanone was obtained by hydrogenation of the natural compound N-[7-(3',4'-methylenedioxyphenyl)-2(Z),4(Z)-heptadienoyl]pyrrolidine. Synthetic amides were prepared by addition of the appropriate amine to the corresponding acyl chloride. The natural compound, N-[7-(3',4'-methylenedioxyphenyl)-2(E),4(E)-heptadienoyl]pyrrolidine, was the most active of all tested compounds against the promastigote and intracellular amastigote forms with IC50 values of 15 µM and 14.5 µM, respectively. None of the compounds modulated the production of nitric oxide.