Chemical variability between different organs of the medicinal plant Casearia sylvestris

The phytochemical profile of essential oils and extracts from Casearia sylvestris leaves, flowers and fruits have been investigated here. Leaf and flower extracts were prepared by sonication and analyzed by thin-layer chromatography and high-performance liquid chromatography. The phenolic content was determined by ultraviolet spectrophotometry. Leaves, flowers, and fruits essential oils were extracted by hydrodistillation. The highest extracts yields were 20.3 % (leaves) and 23.4 % (flowers) with ethanol 70 %. Essential oil extraction yields were 0.3 % (leaves) and 0.1 % (flowers and fruits). Bicyclogermacrene was the major component in all essential oil. Thin-layer chromatography suggests a chemical profile similar for leaves and flowers. The leaves and flowers phenolic content were similar (14.0 and 15.0 %, respectively). Chromatography analyses indicated the predominance of casearin clerodane diterpenes in leaves (λmax 232-235), whereas in flowers, diterpenes with a different standard diene in side-chain C13(16) and C14 (λmax 223-229). The different phytochemical profile of C. sylvestris flowers as compared to the leaves could be explored by the search for new bioactive components. This is the first report on the fruit and flower C. sylvestris essential oil composition. These data could be used as quality control of herbal medicine derived from C. sylvestris leaves.(AU)

Antifungal activities of the essential oil and its fractions rich in sesquiterpenes from leaves of Casearia sylvestris Sw

ABSTRACT Casearia genus (Salicaceae) is found in sub-tropical and tropical regions of the world and comprises about 160-200 species. It is a medicinal plant used in South America, also known as "guaçatonga", "erva-de-tiú", "cafezinho-do-mato". In Brazil, there are about 48 species and 12 are registered in the State of Rio de Janeiro, including Casearia sylvestris Sw. There are many studies related to the chemical profile and cytotoxic activities of extracts from these plants, although few studies about the antifungal potential of the essential oil have been reported. In this work, we have studied the antifungal properties of the essential oil of C. sylvestris leaves, as well as of their fractions, against four yeasts (Saccharomyces cerevisae, Candida albicans, C. glabrata and C. krusei) for the first time. The chemical analysis of the essential oil revealed a very diversified (n = 21 compounds) volatile fraction composed mainly of non-oxygenated sesquiterpenes (72.1%). These sesquiterpenes included α-humulene (17.8%) and α-copaene (8.5%) and the oxygenated sesquiterpene spathulenol (11.8%) were also identified. Monoterpenes were not identified. The fractions are mainly composed of oxygenated sesquiterpenes, and the most active fraction is rich in the sesquiterpene 14-hydroxy -9-epi-β-caryophyllene. This fraction was the most effective in inhibiting the growth of three yeast strains.

Cytotoxic effects of the essential oil from leaves of Casearia sylvestris Sw. (Salicaceae) and its nanoemulsion on A549 tumor cell line / Efectos citotóxicos del aceite esencial de las hojas de Casearia sylvestris Sw. (Salicaceae) y su nanoemulsión sobre líneas celulares tumorales A549

Extracts from leaves of C. sylvestris have cytotoxic effect in different tumor cell lines, possibly due to clerodane type diterpenes (casearins). On the other hand, there are few studies related to the antitumor activity of the essential oils from this species. This work evaluated for the first time the cytotoxicity effects of the pure essential oil and its nanoemulsion against A549 tumor cell line (human lung carcinoma). The essential oil was obtained from fresh leaves by hydrodistillation in a Clevenger-type apparatus and analyzed by GC/MS and GC/FID. Cytotoxicity evaluation was performed using the WST-1 test. The chemical analysis of the essential oil revealed a volatile fraction composed mainly of non-oxygenated sesquiterpenes (72.1%). The essential oil and its nanoemulsion exhibited cytotoxic activity against A549 tumor cells with EC50 of 4.0 µg/mL and EC50 of 1.0 µg/mL, respectively. Both samples displayed a dose dependent pattern (r = -0.79, p = 0.03) as determined by linear regression test.

Los extractos de las hojas de Casearia sylvestris tienen efectos citotóxicos en diferentes líneas celulares tumorales, posiblemente debido a los diterpenos tipo clerodane (casearinas). Por otra parte, hay muy pocos estudios relacionados con la actividad antitumoral del aceite esencial de estas especies. Este trabajo evalúa por primera vez el efecto citotóxico del aceite esencial puro y su nanoemulsión contra la línea de células tumorales A549 (carcinoma humano de pulmón). El aceite esencial fue obtenido de hojas frescas por hidrodestilación en un aparato tipo Clevenger y analizado por GC/MS y GC/FID. La evaluación de citotoxicidad fue realizada usando la prueba WST-1. El análisis químico del aceite esencial reveló una fracción volátil compuesta principalmente por sesquiterpenos no oxigenados (72,1%). El aceite esencial y su nanoemulsiónexhibió actividad citotóxica contra las células tumorales A549 con una EC50 de 4,0 µg/mL y una EC50 de 1,0 µg/mL, respectivamente. Ambas muestras exhibieron un patrón dosis-dependiente (r = -0,79, p = 0,03) determinado por análisis de regresión lineal.

Anatomical aspects, chemical analysis and cytotoxic effect of the essential oil from leaves of Casearia arborea (Salicaceae) / Aspectos anatómicos, análisis químico y efecto citotóxico del aceite esencial de Casearia arborea (Salicaceae)

The genus Casearia (Salicaceae) is found in sub-tropical and tropical regions of the world and comprises about 200 species. In Brazil, there are about 48 species and 12 are registered in the State of Rio de Janeiro; including Casearia arborea (Rich.) Urb. Essential oil was obtained from the fresh leaves by hydrodistillation and analyzed by GC-MS and GC-FID. The cytotoxic effect was determined by WST-1 assay. Chemical analysis of the essential oil revealed a very diversified (n = 37 compounds) volatile fraction composed mainly of non-oxygenated sesquiterpenes (90.2%). These sesquiterpenes included byciclogermacrene (18.7%), germacrene D (12.1%) and α-humulene (11.5%). In addition, the essential oil demonstrated cytotoxic effects against A549 tumor cells in the concentration of 4 µg/mL (EC50) (p < 0.05).

El género Casearia (Salicáceas) se encuentra en las regiones tropicales y sub-tropicales del planeta y comprende alrededor de 200 especies. En Brasil existen 48 especies, 12 de las cuales fueron registradas en el Estado de Río de Janeiro incluyendo Casearia arborea (Rich.) Urb. El aceite esencial fue extraído de hojas frescas por hidrodestilación y analizado por GC-MS y GC-FID. El efecto citotóxico fue determinado por ensayo WST-1. Las cavidades secretorias fueron ocasionalmente encontradas tanto en la lámina foliar como en el pecíolo. El análisis químico del aceite esencial reveló una muy diversa fracción volátil (n = 37 compuestos) formada principalmente por sesquiterpenos no oxigenados (90,2%). Estos sesquiterpenos incluyen biciclogermacreno (18,7%), germacreno D (12,1%) y α-humuleno (11,5%). Además, el aceite esencial demostró efectos citotóxicos contra las células tumorales A549 en una concentración de 4µg/mL (EC50) (p < 0.05).

Antimicrobial potential of Casearia sylvestris against oral bacteria / Potencial antimicrobiano da Casearia sylvestris frente a bactérias orais

Aim: The aim of this study was to obtain Casearia sylvestris leave extracts by different extractive methods, including the obtention of essential oil, in order to compare their antimicrobial activities to conventional mouthwash chlorhexidine against oral bacteria. Material and method: For this evaluation, extracts from the leaves were obtained by different methods of extraction (infusion, decoction, maceration and percolation) using different solvent systems: water 100%, ethanol 100%, methanol 100%, water: ethanol 3:7; water: ethanol 7:3; water: methanol 7:3 and water: methanol 3:7. The essential oil, which corresponds to a volatile fraction, was obtained by hydrodistillation using Clevenger modified apparatus. The microdilution broth method was used to determine the values of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for the following microorganisms: Streptococcus mutans ATCC 25175, S. mitis ATCC 49456, S. sanguinis ATCC 10556, S. salivarius ATCC 25975, Lactobacillus casei ATCC 11578 and Enterococcus faecalis ATCC4082. Chlorhexidine gluconate was used as a positive control. Result: All extracts evaluated in the used protocol displayed MIC values higher than 400 µg/mL and few showed bactericidal activity. The antimicrobial activity of essential oil was higher than the activity of the extracts, and the best minimum inhibitory concentration and minimum bactericidal concentration values were obtained against L. casei (MIC of 0.023 µg/mL and MBC of 0.046 µg/mL) and S. mutans (MIC of 25 µg/mL and MBC of 50 µg/mL), respectively. Conclusion: The essential oil of Casearia sylvestris has significant antimicrobial activity against oral microorganisms.

Objetivo: O objetivo deste estudo foi obter extratos por diferentes métodos de extração de Casearia sylvestris, incluindo a obtenção de óleo essencial, a fim de comparar suas atividades antimicrobianas em relação aos enxaguatórios bucais convencionais com clorexidina contra bactérias orais. Material e método: Para esta avaliação, extratos das folhas foram obtidos por diferentes métodos de extração (infusão, decocção, maceração e percolação) utilizando diferentes sistemas de solventes: 100% de água, 100%, de etanol, metanol 100%, água: etanol 3: 7; água: etanol 7: 3; água: metanol 7: 3 e água: metanol 3: 7. O óleo essencial, que corresponde a uma fração volátil, foi obtido por hidrodestilação usando o aparelho de Clevenger modificado. O método de microdiluição em caldo foi usado para determinar os valores de concentração inibitória mínima (MIC) e a concentração bactericida mínima (MBC) para os seguintes microrganismos: Streptococcus mutans ATCC 25175, ATCC 49456 S. mitis, S. sanguinis ATCC 10556, S. salivarius ATCC 25975, Lactobacillus casei ATCC 11578 e Enterococcus faecalis ATCC4082. Gluconato de clorexidina foi usada como um controle positivo. Resultado: Todos os extratos avaliados no protocolo utilizado apresentaram valores de MIC superior a 400 ug/mL e alguns mostraram atividade bactericida. A atividade antimicrobiana do óleo essencial foi maior do que a atividade dos extratos, e a melhor concentração inibitória mínima e valores de concentração bactericida mínima foram obtidos contra L. casei (MIC de 0,023 ug/mL e MBC de 0,046 ug/mL) e S. mutans (MIC de 25 ug/mL e MBC de 50 ug /mL), respectivamente. Conclusão: O óleo essencial de Casearia sylvestris tem atividade antimicrobiana significativa contra microrganismos orais.

Analysis of the reaction of subcutaneous tissues in rats and the antimicrobial activity of calcium hydroxide paste used in association with different substances

The aim of this study was to evaluate the subcutaneous tissue response in rats and the antimicrobial activity of intracanal calcium hydroxide dressings mixed with different substances against E. faecalis. Fifty four rats were divided into three experimental groups according to the vehicle in the calcium hydroxide treatment: 0.4% chlorohexidine in propylene glycol (PG),Casearia sylvestris Sw in PG and calcium hydroxide+PG (control group). The pastes were placed into polyethylene tubes and implanted into the subcutaneous tissue. After 7, 14 and 30 days, the samples were processed and histologically evaluated (hematoxylin and eosin). The tissue surface in contact with the material was analyzed, and the quantitative analysis determined the volume density occupied by the inflammatory infiltrate (giant cells, polymorphonuclear cells and mononuclear cells), fibroblasts, collagen fibers and blood vessels. For the antimicrobial analysis, 20 dentin blocks infected with E. faecalis were treated with calcium hydroxide pastes in different vehicles; 0.4% chlorhexidine in PG, PG, extract fromCasearia sylvestris Sw in PG and a positive control (infection and without medication) for 7 days. The efficiency of the pastes was evaluated by the live/dead technique and confocal microscopy. The results showed that 0.4% chlorhexidine induced a higher inflammatory response than the other groups. The Casearia sylvestris Sw extract showed satisfactory results in relation to the intensity of the inflammatory response. In the microbiological test, there were no statistical differences between the evaluated intracanal dressings and the percentage of bacterial viability was between 33 and 42%. The control group showed an 86% viability. Antimicrobial components such as chlorhexidine or Casearia sylvestris Sw did not improve the antimicrobial activity against E. faecalis in comparison to the calcium hydroxide+PG treatment. In addition, the incorporation of chlorhexidine in the calcium hydroxide paste promoted the highest inflammatory response.

Casearia sylvestris Swartz Extract Release Using Natural Rubber Latex Biomembranes as Carrier.

The Natural Rubber Latex (NRL) from Hevea brasiliensis has shown promise in biomedical applications due to its low cost, easy handling, mechanical properties and biocompatibility, being used for bone regeneration and wound healing due to its natural stimulus to angiogenesis. The aim of this work was to incorporate Casearia sylvestris Sw. extract in NRL biomembranes and study its release behavior. The complex membraneextract has as object of study a new approach of using C. sylvestris extract in the treatment of wounds, for possessing antiseptic activity, anti-inflammatory and analgesic properties. The C. sylvestris species (Salicaceae), popularly known as "guaçatonga", presents great distribution and is used in folk medicine as antiulcer, wound healing, anti- snake venom, properties which have been proven and related to clerodane diterpenes (casearins A-X). The release rate of C. sylvestris compounds from extract-membrane complex was monitored and analyzed using the method of optical spectroscopy (UV-VIS). The release varied with temperature ranging from 14 to 33 days, releasing more than 90%, with an interesting and promising biomedical application, such as wound healing and burns.

Aspectos toxicológicos da planta medicinal Casearia sylvestris Swartz: revisão de literatura / Toxicology of the medicinal plant Casearia sylvestris Swartz: a literature review

In the West, the consumption of medicinal plants is increasing strongly, and up to 40% of the population use medicinal plants and phytotherapic preparations regularly, in the belief that they are innocuous and/or safe. However, in most cases there is no scientific proof of their pharmacological and toxicological properties. The aim of this study was to carry out a bibliographical review of the toxicological properties of Casearia sylvestris Swartz and collect data providing a basis for its rational use. Some studies validate classical therapeutic indications of C. sylvestris, such as to treat diarrhea and snakebites, justifying its empirical use. Nevertheless, there are few reports about its toxicological properties and these present only limited findings, describing results as 50% lethal dose (LD50) and the acute toxicity of its constituents. Currently, research is insufficient to ensure safety of popular preparations based on C. sylvestris. Therefore, it is necessary to make a fuller assessment of its deleterious profile, especially with respect to the toxic potential of its constituents, as well as its capacity to harm target organs and organic systems...

No Ocidente, o consumo de plantas medicinais vem aumentando substancialmente, onde cerca de 40% da população utiliza regularmente plantas medicinais e preparações fitoterápicas sob o rótulo de serem produtos inócuos ou seguros. Porém, na maioria das vezes, não há comprovação científica de suas propriedades farmacológicas e toxicológicas. O objetivo deste trabalho foi realizar um levantamento bibliográfico sobre as propriedades toxicológicas da Casearia sylvestris Swartz e coletar dados que fundamentem seu uso racional. Alguns estudos validam indicações terapêuticas clássicas da C. sylvestris como antidiarréico e no tratamento de ferimentos ofídicos, fundamentando seu uso empírico. Porém, poucos trabalhos relatam suas propriedades toxicológicas, havendo apenas abordagens limitadas em torno da Dose Letal 50% e da toxicidade aguda de seus constituintes. Até o presente momento, as pesquisas são insuficientes para garantir a segurança de preparações populares à base de C. sylvestris. Portanto, há necessidade de avaliar melhor seu perfil deletério, principalmente, no que diz respeito ao potencial tóxico de seus constituintes, assim como a sua capacidade lesiva sobre órgãos-alvos e sistemas orgânicos...

Anti-Escherichia coli activity of Brazilian plant extracts: New trends in Veterinary research / Atividade anti-Escherichia coli de extratos de plantas brasileiras: Novas tendências em pesquisa veterinária

Escherichia coli é uma bactéria presente no trato intestinal de animais de sangue quente. Entretanto, a perda do equilíbrio em sua relação com o hospedeiro pode causar doenças que levam a perdas econômicas, quando se trata de animais de produção, e perdas incalculáveis, quando se trata de animais silvestres. O presente estudo testou diversos extratos aquosos e orgânicos, obtidos de plantas da Amazônia brasileira, contra Escherichia coli (ATCC 25922). Usando o ensaio de disco difusão em ágar Mueller-Hinton (DDA), foram testados 1.791 extratos quanto à sua atividade antibacteriana. Os extratos que mostraram halo de inibição de crescimento bacteriano foram testados no modelo da microdiluição em caldo (MDBC) contra uma suspensão bacteriana preparada na escala de 0,5 McFarland para determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM). Dois extratos orgânicos (EB 127 e EB725) e dois extratos aquosos (EB 272 e EB 934) testados inibiram o crescimento bacteriano no modelo DDA. O extrato orgânico EB127, obtido de Microplumeria sp., apresentou CIM e CBM≥700mg/mL; o extrato aquoso EB272, obtido de Casearia sp., apresentou CIM=CBM=600mg/mL; o extrato orgânico EB725, obtido de Buchenavia sp., apresentou CIM=CBM<400mg/mL; e o extrato aquoso EB934, obtido de Caryocar sp., apresentou 400mg/mL

Efecto in vitro de los terpenos lupeol y casearina G sobre células sanguíneas y tumorales / In vitro effect of lupeol and casearin G on peripheral blood mononuclear and tumor cells

Background: The rainforest is an important source of natural compounds with therapeutic properties. Although there are many anti-inflammatory and antineoplastic drugs available to the clinician, there is an ongoing need for new therapeutic drugs with fewer serious adverse effects. Aim: To evaluate the in vitro cytotoxic effects of lupeol and casearin G on tumor cells, on phagocytic activity and nitric oxide (NO) production by blood mononuclear cells. Material and Methods: The cytotoxic effect of these compounds on cell lines MCF-7 (human breast adenocarcinoma) and PC-3 (human prostate cancer) was measured by a colorimetric assay (MTS/PMS) and the sulphorhodamine B assay. Peripheral blood mononuclear cells were obtained from eight healthy volunteers. The effect of these compounds on nitric oxide (NO) production was measured using the Griess reaction. Their effect on phagocytic activity of PBMC was also evaluated. Results: Lupeol (≥ 2 mM) resulted in a reduction of both the phagocytic index and the percentage of phagocytic monocytes and macrophages. Treatment of monocytes/macrophages with lupeol (72 µM) and casearin G (4 µM) reduced the production of NO. Neither lupeol (< 969 µM) nor casearin G (< 55 µM) had cytotoxic effects on PBMC. Casearin G showed both cytotoxic (IC50, LC50) and cytostatic (GI50) effects against tumor cells, PC-3 (IC50 = 12.5 µM; GI50 = 13.3 µM; LC50 = 51.9 µM) and MCF-7 (IC50 = 112.8 µM; GI50 = 11.8 µM; LC50 = 49.4 µM), as well as a hemolytic effect (≥ 182 µM). Conclusions: These observations indicate that lupeol and casearin G might be useful compounds in the preparation of anti-inflammatory drugs, whereas casearin G might be useful in the elaboration of antitumor drugs.

Folk uses and pharmacological properties of Casearia sylvestris: a medicinal review

Folk uses and scientific investigations have highlighted the importance of Casearia sylvestris extracts and their relevant bioactive potential. The aim of this work was to review the pharmacological properties of C. sylvestris, emphasizing its anti-ulcer, anti-inflammatory, anti-ophidian and antitumor potentialities. Ethanolic extracts and essential oil of their leaves have antiulcerogenic activity and reduce gastric volume without altering the stomach pH, which corroborates their consumption on gastrointestinal disorders. Leaf water extracts show phospholipase A2 inhibitory activity that prevents damage effects on the muscular tissue after toxin inoculation. This antiphospholipasic action is probably related to the use as an anti-inflammatory, proposing a pharmacological blockage similar to that obtained with non-steroidal anti-inflammatory drugs on arachidonic acid and cyclooxygenase pathways. Bioguided-assay fractionations lead to the identification of secondary metabolites, especially the clerodane diterpenes casearins (A-X) and casearvestrins (A-C), compounds with a remarkable cytotoxic and antitumor action. Therefore, the C. sylvestris shrub holds a known worldwide pharmacological arsenal by its extensive folk utilization, exciting searches for new molecules and a better comprehension about biological properties.

Usos populares e pesquisas científicas têm destacado a importância dos extratos da planta Casearia sylvestris e seu grande potencial bioativo. Neste trabalho, objetiva-se revisar as propriedades farmacológicas de C. sylvestris, enfatizando sua potencialidade antiulcerogênica, antiinflamatória, antiofídica e antitumoral. O extrato etanólico e o óleo essencial das folhas possuem atividade antiulcerogênica promissora, diminuindo o volume gástrico sem alterar o pH estomacal, corroborando sua aplicação contra dores gastrointestinais. Já os extratos aquosos das folhas têm atividade inibitória contra fosfolipase A2 presente em venenos de cobras, atenuando os efeitos lesivos sobre a musculatura esquelética resultantes da inoculação das toxinas. Essa ação antifosfolipásica provavelmente está relacionada ao seu uso como antiinflamatório, sugerindo um bloqueio análogo ao dos fármacos antiinflamatórios não-esteroidais na formação de mediadores oriundos do ácido araquidônico e na ativação da ciclooxigenase. Ensaios de fracionamento bioguiado dos extratos culminaram no isolamento e identificação de inúmeros metabólitos secundários, especialmente os diterpenos clerodânicos casearinas (A-X) e casearvestrinas (AC), compostos que têm surpreendido por sua ação citotóxica e antitumoral. Assim, a planta C. sylvestris apresenta um enorme arsenal farmacológico já mundialmente comprovado por seu vasto uso popular, estimulando pesquisas por novas moléculas e a busca pela compreensão de suas propriedades biológicas.

Determinação in vitro do efeito antimicrobiano direto do hidróxido de cálcio associado a diferentes substâncias frente a cepas de Enterococcus faecalis / In vitro determination of direct antimicrobial effect of calcium hydroxide associated with different substances against Enterococcus faecalis strains

Objetivo: determinar o efeito antimicrobiano em contato direto de Casearia sylvestris Swart (guaçatonga), propilenoglicol,e da clorexidina associada ao hidróxido de cálcio contra 40 cepas de Enterococcus faecalis isoladas da cavidade bucal. Métodos: após a ativação, as cepas bacterianas foram suspensas em solução salina estéril até a escala 1,0 de MacFarland. A suspensão foi colocada em contato direto com hidróxido de cálcio [Ca(OH)2] + propilenoglicol puro, Ca(OH)2 + clorexidina a 1% em propilenoglicol, e Ca(OH)2 + extrato de guaçatonga em propilenoglicol, cobrindo pontas de papel, previamente contaminadas com a solução por 3 minutos, com as diferentes pastas. A atividade antimicrobiana foi avaliada em 6, 24, 48, 72 horas, e aos 7 dias. Após o período de incubação, os cones foram retirados das pastas e incubados em caldo Letheen a 37ºC por 48 horas. Em seguida, 0,1 ml do caldo Letheen foi transferido para tubos contendo infusão de cérebro e coração(BHI) e incubados em caldo novamente a 37ºC por 48 horas.A turbidez foi observada no meio. Depois disso, placas de ágar M-Enterococcus foram semeadas com caldo BHI de cada tubo e o crescimento colonial foi avaliado. Resultados: todas as cepas bacterianas foram inibidas por todas as pastas, nos períodos avaliados. Conclusões: concluiu-se que a adição dessas substâncias ao hidróxido de cálcio não interferiu no seu efeito antimicrobiano direto.

Objective: To determinate the direct antimicrobial effectsof Casearia sylvestris Swart (guaçatonga), propylene glycol,and of chlorhexidine associated to calcium hydroxidepaste against 40 Enterococcus faecalis strains isolated fromthe oral cavity when direct contact. Methods: After activation,the bacterial strains were suspended in sterile salineto 1.0 McFarland standard. The suspension was placedin direct contact with calcium hydroxide paste [Ca(OH)2]+ pure propylene glycol, Ca(OH)2 + chlorhexidine 1%in propylene glycol, and Ca(OH)2 + guaçatonga extractin propylene glycol by covering paper points, previouslycontaminated for 3 minutes, with the different pastes. Antimicrobialactivity was evaluated at 6, 24, 48, 72 hours,and at 7 days. After the incubation period, the points wereremoved from the pastes and incubated in Letheen brothat 37oC for 48 hours. Following that, 0.1ml of the Letheenbroth was transferred to tubes containing brain heart infusion(BHI) broth and incubated again at 37oC for 48hours. Turbidity was observed in the medium. After that,M-Enterococcus agar plates were seeded with BHI brothfrom each tube and colony growth was assessed. Results:All the bacterial strains were inhibited by all pastes at theevaluated periods. Conclusions: It was concluded that theaddition of these substances to calcium hydroxide did notinterfere with its direct antimicrobial effect.

Bothrops jararacussu venom-induced neuromuscular blockade inhibited by Casearia gossypiosperma briquet hydroalcoholic extract

The hydroalcoholic extract of Casearia gossypiosperma Briquet (Flacourtiaceae) was standardized for the first time through quality control procedures including pharmacognostic methods, fingerprint chromatograms, defined amounts of marker substances and physicochemical characteristics. The pharmacological activity of C. gossypiosperma (Cg) hydroalcoholic extract was assayed by a traditional in vitro test, which involved irreversible neuromuscular blockade induced by Bothrops jararacussu (Bjssu) venom (60 µg/mL) in mouse phrenic nerve-diaphragm preparations. Bjssu venom blocked muscle activity for 26 (± 2.0) minutes (n = 6). Cg extract (0.1 mg/mL) induced changes on the baseline muscle activity without impairing the muscle function and inhibited 87.6 percent (± 1.8) (n = 6) of the Bjssu venom-induced blockade. Both flavonoids (0.624 g percent) and polyphenols (4.63 g percent) from the extract were spectrophotometrically quantified. Therefore, the present study confirms the antibothropic activity of Cg extract, supporting the ethnomedical use of Casearia sp. in the treatment of snakebite victims.

Bothrops jararacussu venom-induced neuromuscular blockade inhibited by Casearia gossypiosperma Briquet hydroalcoholic extract

The hydroalcoholic extract of Casearia gossypiosperma Briquet (Flacourtiaceae) was standardized for the first time through quality control procedures including pharmacognostic methods, fingerprint chromatograms, defined amounts of marker substances and physicochemical characteristics. The pharmacological activity of C. gossypiosperma (Cg) hydroalcoholic extract was assayed by a traditional in vitro test, which involved irreversible neuromuscular blockade induced by Bothrops jararacussu (Bjssu) venom (60 ìg/mL) in mouse phrenic nerve-diaphragm preparations. Bjssu venom blocked muscle activity for 26 (± 2.0) minutes (n = 6). Cg extract (0.1 mg/mL) induced changes on the baseline muscle activity without impairing the muscle function and inhibited 87.6% (± 1.8) (n = 6) of the Bjssu venom-induced blockade. Both flavonoids (0.624 g%) and polyphenols (4.63 g%) from the extract were spectrophotometrically quantified. Therefore, the present study confirms the antibothropic activity of Cg extract, supporting the ethnomedical use of Casearia sp. in the treatment of snakebite victims.

Identificação de diterpenos clerodânicos em diferentes órgãos de Casearia sylvestris Swartz / Identification of clerodane diterpenes in different organs of Casearia sylvestris Swartz

Casearia sylvestris Swartz (Salicaceae) é uma espécie vegetal arbórea ou arbustiva, disseminada pelo território brasileiro e bastante utilizada em nossa medicina popular. Diversos diterpenos clerodânicos bioativos típicos do gênero Casearia foram isolados desta espécie (ex. casearinas e casearvestrinas). O objetivo principal deste trabalho foi avaliar a presença de diterpenos clerodânicos em diferentes órgãos de C. sylvestris, utilizando técnicas analíticas cromatográficas e espectroscópicas. Os extratos dos diferentes órgãos foram analisados por cromatografia em camada delgada, cromatografia líquida de alta eficiência com detector de arranjo de diodos e ressonância magnética nuclear de 1H. Nas análises cromatográficas foram utilizados diterpenos clerodânicos isolados de C. sylvestris como padrões, incluindo rel-19S-acetóxi- 18R- butanoilóxi- 18,19- epóxi -6S -hidróxi -2R- (2-metilbutanoilóxi) -5S, 8R, 9R, 10S -cleroda-3,13(16),14-trieno, isolado do caule pela primeira vez. Foram obtidos perfis fitoquímicos dos órgãos avaliados. Os resultados indicaram a presença de diterpenos clerodânicos em toda planta, prevalecendo em maior número nas folhas. Além disso, os resultados também indicam que em caules, flores e raízes há predomínio de diterpenos clerodânicos com padrão diênico diferente daqueles obtidos de folhas de C. sylvestris.

Casearia sylvestris Swartz (Salicaceae) is a tree or shrub distributed widely in Brazil, where it is used in popular medicine. Several bioactive clerodane diterpenes typical of Casearia have been isolated from this species (e.g. casearins and casearvestrins). The main objective of this study was to identify clerodane diterpenes in various organs of C. sylvestris, using chromatographic and spectroscopic analytical techniques. The extracts of the different plant parts were analyzed by thin layer chromatography, high performance liquid chromatography with diode array detector and 1H nuclear magnetic resonance. In the chromatographic analysis, clerodane diterpenes isolated from C. sylvestris were used as standards, including rel-19Sacetóxi- 18R- butanoilóxi-18,19- epóxi -6S -hidróxi -2R- (2-metilbutanoilóxi) -5S, 8R, 9R, 10S -cleroda-3,13(16),14- triene, isolated for the first time from the stems. Phytochemical profiles of the organs were produced, which indicated the presence of clerodane diterpenes in all parts of the plant, notably in the leaves. The results also suggest that the main clerodane diterpenes in the stems, flowers and roots had conjugated double-bond patterns that differed from those found in the leaves.

Assessment of the antimicrobial activity of Casearia sylvestris extract against oral pathogenic microorganisms

An ethanolic extract of leaves from the tree Casearia sylvestris, known as guaçatonga in Brazil, was tested for in vitro activity against oral pathogenic bacteria and fungi. The results showed susceptibility of all the microorganisms tested. This study suggests a potential use of ethanolic extract of C. sylvestris as a novel treatment of oral infectious conditions, such as denture stomatitis, periodontitis and dental caries.

Comparação da atividade antimicrobiana in vitro de diferentes antibióticos e extratos hidroalcoólicos e infusão de casearia sylvestris swart (guaçatonga) frente a linhagens de enterococcus faecalis isolados da cavidade oral / Comparison of in vitro antimicrobial activity of different antibiotics, hydroalcoholic extracts and infusion of casearia sylvestris swart (guacatonga) against strains of Enterococcus faecalis isolated from the oral cavity

Enterococcus faecalis são cocos Gram positivos, elipsóides, em cadeias curtas, anaeróbios facultativos, habitantes dos tratos intestinal e genital e da cavidade oral de humanos e animais. Podem causar uma gama muito grande de doenças no homem como cistite, endocardite e infecções de feridas. Os enterococos têm sido implicados também em infecções endodônticas, comumente detectados em canais radiculares obturados exibindo sinais de periodontite apical crônica e lesões refratárias periapicais. É de preocupação do cirurgião dentista um completo saneamento do sistema de canais radiculares pela aplicação de medicação antisséptica entre sessões. Diante desta problemática, este estudo comparou a atividade antimicrobiana de extratos alcoólicos e aquoso da Casearia sylvestris Sw (Guaçatonga) com a atividade antimicrobiana da amoxicilina, azitromicina, vancomicina, penicilina, clindamicina, cloranfenicol e da clorexidina frente a 50 linhagens de E. faecalis isolados da cavidade oral de pacientes da Clínica de Endodontia da USC. Os testes foram realizadospelo método de difusão das drogas a partir de discos impregnados com as substâncias sobre a superfície de placas de ágar Mueller-Hinton. A droga de melhor eficiência foi amoxicilina com 100% das linhagens sensíveis, estatisticamente signifi cativo (p<0,05) quando comparadas às outras drogas. A droga de menor eficiência foi a clindamicina com 94% de linhagens resistentes. Dos extratos obtidos daplanta, o extrato propilenoglicólico mostrou melhor efi ciência, inibindo34 (68%) das estirpes (média de 4,31mm de halo de inibição). A clorexidina inibiu 100% das estirpes (média de 17,09mm de halo de inibição) (CNPq).

Cytotoxic evaluation of essential oil from Casearia sylvestris Sw on human cancer cells and erythrocytes / Avaliação citotóxica do óleo essencial de Casearia sylvestris Sw sobre células tumorais humanas e eritrócitos

Casearia sylvestris (Flacourtiaceae) is a plant popularly known as "guaçatonga" and it is used by indigenous population from South America (Brazil, Peru and Bolivia) in the treatment of several diseases, including cancer. Cytotoxic studies showed that it presents an interesting antitumoral potential due to the presence of casearins and the essential oil showed a high percentage of potent cytotoxic sesquiterpenes (β-caryophyllene and α-humulene). In this work, we verified that the essential oil of C. sylvestris presents a good selective cytotoxicity against HeLa, A-549 and HT-29 tumor cells (CD50 63.3, 60.7 and 90.6 µg.ml-1, respectively) when compared to non-tumoral cells Vero (CD50 210.1 µg.ml-1) and mice macrophages (CD50 234.0 µg.ml-1). The oil causes hemolysis in seven different kinds of erythrocytes, indicating that C. sylvestris must be used carefully. Besides, standard of β-caryophyllene and α-humulene were also tested and they showed similar cytotoxicity to the cytotoxicity presented in the oil, indicating that they might be responsible for the toxic effects that were observed in this study.

A Casearia sylvestris (Flacourtiaceae) é uma planta popularmente conhecida como "guaçatonga" e é usada por povos indígenas da América do sul (Brasil, Peru e Bolivia) no tratamento de muitas doenças, incluindo câncer. Estudos citotóxicos mostraram que esta planta apresenta um possível e interessante potencial antitumoral devido à presença de moléculas chamadas casearinas. Além disso, a composição do óleo essencial mostrou uma alta concentração de sesquiterpenos de alto potencial citotóxico. Neste trabalho, nós verificamos que o óleo essencial da C. sylvestris apresentou uma boa citotoxicidade seletiva contra as linhagens de células tumorais HeLa, A-549 and HT-29 (CD50 63,3, 60,7 e 90,6 µg.ml-1, respectivamente) quando comparada às células não-tumorais Vero (CD50 210,1 µg.ml-1) e macrófagos de camundongos (CD50 234,0 µg.ml-1). Além disso, o óleo causou hemólise em sete diferentes tipos de eritrócitos, indicando que a C. sylvestris precisa ser usada com cuidado. Também foram testados padrões de β-cariofileno e α-humuleno que mostraram citotoxicidade similar àquelas apresentadas pelo óleo, indicando que estes compostos podem ser os responsáveis pelos efeitos tóxicos que foram observados neste estudo.

Comparação da casearia silvestris com indometacina e placebo na diminuição de edema em pata traseira de Ratos Wistar, induzido pelo uso de carragenina / Comparison of silvestris caesarean with indometacina and placebo on the reduction of back legãs edema of Wistar Rats, induced by carragenin use

O experimento visa verificar e comparar a diminuiçãodo edema de pata traseira em ratos Wistar (previamenteinduzido pela droga inflamatória carragenina), como uso de Casearia silvestris, de indometacina e de solução de Tween a 2% (placebo). Na realização deste estudo experimental, os ratos foram subdivididos em três grupos que receberam as injeções com a solução placeboe drogas antiinflamatórias uma hora antes da induçãodo edema de pata, previamente descrita. Após seis períodos, com verificações espaçadas de 30 minutos, comparou-se o comportamento do edema da pata de cada grupo. Obteve-se como resultado uma progressão do volume do edema até 240 minutos, no grupo em que foi administrado a solução sem nenhum agente antiinflamatório, com uma elevação média percentual do volume de 4,5ml. No grupo da administração da indometacina verificou-se diminuição considerável do volume após 2 horas e com elevação média de 0,75. No grupo da Casearia, o edema teve redução menor que da indometacina, com uma média do aumento de volume de 3ml, porém, superior ao placebo. Assim, verificou-se que houve uma diminuição do edema com as drogas antiinflamatórias, principalmente com a indometacina...

Toxicidade aguda e propriedades antiofídicas do extrato aquoso de Casearia grandiflora (Flacourtiaceae): atividades fosfolipásica A2, miotóxica e letal de peçonha de B. moojeni e B. neuwiedi / Acute toxicity and biological activities of Casearia grandiflora aqueous extract (Flacourtiaceae): phospholipase A2, myotoxic and lethal activities of venoms of B. moojeni and B. neuwiedi

O objetivo do presente trabalho foi estudar a ação do extrato aquoso de C. grandiflora (EA) sobre as atividades PLA2 miotóxica e letal das peçonhas (P) de B. moojeni e B. neuwiedi e sua toxicidade aguda. O EA (1:160; P:EA, m/m) inibiu em 74,5 por cento e 57,5 por cento a atividade PLA2 das peçonhas e, em 51 por cento, a atividade CK do plasma de camundongos induzida pela peçonha de B. moojeni na proporção de 1:4 (P:EA, m/m). Houve um aumento da sobrevivência (83 por cento) dos camundongos que receberam EA preparado com folhas coletadas em novembro (1:26; m/m), resultado não encontrado com o EA preparado a partir de folhas coletadas em junho. A efeitos colaterais do EA de C. grandiflora foram ptose palpebral, letargia e piloereção e, apnéia, paralisia flácida e óbito (100 por cento), respectivamente para as doses de 250 e 500 mg. Kg-1. Estes resultados indicam que o EA é uma fonte de compostos capazes de neutralizar alguns efeitos tóxicos de peçonhas botrópicas, porém, investigações adicionais são necessárias para eliminar ou minimizar seus efeitos colaterais.