Brine shrimp (Artemia salina Leach) bioassay of extracts from Lychnophoriopsis candelabrum and different Lychnophora species

O presente estudo teve como objetivo uma citotoxicidade sobre Artemia salina de vinhos e dois extratos de cinco espécies do gênero Lychnophora e de uma espécie de Lychnophoriopsis . Os extratos solubilizados em DMSO, preparados nas concentrações finais de 100, 250, 375, 500 e 600 µg mL -1 , foram adicionados a recipientes contendo náuplios de Artemia salina (10 unidades cada) e completo em volume para 5 mL de solução marinha. Lapachol e DMSO 5% foram como controles positivo e negativo, respectivamente. Como amostras foram mantidas sob iluminação e como larvas mortas foram contadas após 24 horas de contato. O cálculo da LC 50 foi feito com o programa Probitos. Os extratos brutos etanólicos de cinco Espécies apresentaram baixa letalidade nd Seguinte Ordem: Lychnophora trichocarpha (LC 50 = 672,38 ng mL -1 )> Lychnophora pinaster (LC 50 = 678,73 ng mL -1 )> Lychnophora ericoides (LC 50 = 738,09 µg mL -1 )> Lycellophoriopsis candelabro (LC 50 = 812,57 µg mL -1 )> Lychnophora passerina (LC 50 = 921,78 µg mL -1 ). Todos os extratos testados de Lychnophoriopsis candelabro eo extrato clorofórmico de Lychnophora staavioides mostraram leve toxicidade sobre A. salina . Os resultados indicaram que existem substâncias com potencial atividade farmacológica em todas as espécies testadas.

The present study aimed to evaluate on Artemia salina the citotoxicity of twenty-two extracts from five species of the genus Lychnophora and one species of the genus Lychnophoriopsis. The extracts solubilized in DMSO and prepared at the final concentrations of 100, 250, 375, 500 and 600 g mL-1 were added to tubes containing Artemia salina nauplii (10 units each) and filled to 5 mL total volume with artificial salt water. Lapachol and 5% DMSO were used as positive and negative controls, respectively. The samples were kept under light and dead larvae were counted after 24 hours of contact. LC50 was calculated by using Probit software. The crude ethanol extracts from five species showed low lethality in the following order: Lychnophora trichocarpha (LC50 = 672.38 g mL-1) > Lychnophora pinaster (LC50 = 678.73 g mL-1) > Lychnophora ericoides (LC50 = 738.09 g mL-1) > Lychnophoriopsis candelabrum (LC50 = 812.57 g mL-1) > Lychnophora passerina (LC50 = 921.78 g mL-1). All tested extracts from L. candelabrum and chloroform extract from L. staavioides showed light toxicity on A. salina. Results indicated that there are substances with potential pharmacological activity in all tested species

Anti-inflammatory activity of Lychnophora passerina, Asteraceae (Brazilian "Arnica").

ETHNOPHARMACOLOGICAL RELEVANCE: Lychnophora passerina (Asteraceae), popularly known as "arnica," is used to treat inflammation, pain, rheumatism, contusions, bruises and insect bites in Brazilian traditional medicine. MATERIALS AND METHODS: The anti-inflammatory activity of crude ethanolic extract of aerial parts of L. passerina and its ethyl acetate and methanolic fractions had their abilities to modulate the production of NO, TNF-α and IL-10 inflammatory mediators in LPS/IFN-γ-stimulated J774.A1 macrophages evaluated. Moreover, the crude ethanolic extract and derived fractions were also in vivo assayed by carrageenan-induced paw oedema in mice. RESULTS: In vitro assays showed remarkable anti-inflammatory activity of L. passerina crude ethanolic extract (EE) and its ethyl acetate (A) and methanolic (M) fractions, through the inhibition of production of NO and TNF-α inflammatory mediators and induction of production of IL-10 anti-inflammatory cytokine. In vivo assays showed anti-inflammatory activity for EE 10% ointment, similar to the standard drug diclofenac gel. The A and M fraction ointments 20% presented anti-inflammatory activity. CONCLUSION: The results obtained showed that possible anti-inflammatory effects of EE and its A and M fractions may be attributed to inhibition pro-inflammatory cytokines production, TNF-α and NO and to increased IL-10 production. EE, A and M ointments showed topical in vivo anti-inflammatory activity. The in vivo anti-inflammatory activity of EE of L. passerina may be related to synergistic effects of different substances in the crude extract. Therefore, traditional use of aerial parts of L. passerina in the inflammatory conditions could be beneficial to treat topical inflammatory conditions, as evidenced by the present study.

Antiviral activity of Bignoniaceae species occurring in the State of Minas Gerais (Brazil): part 1.

AIMS: To evaluate the antiviral activity of Bignoniaceae species occurring in the state of Minas Gerais, Brazil. METHODS AND RESULTS: Ethanol extracts of different anatomical parts of bignoniaceous plant species have been evaluated in vitro against human herpesvirus type 1 (HSV-1), vaccinia virus (VACV) and murine encephalomyocarditis virus (EMCV) by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. A total of 34 extracts from 18 plant species selected according to ethnopharmacological and taxonomic criteria were screened. Fifteen of the 34 extracts (44.1%) have disclosed antiviral activity against one or more of the viruses assayed with EC(50) values in the range of 23.2 ± 2.5-422.7 ± 10.9 µg ml(-1). CONCLUSIONS: Twelve of the 34 extracts (35.3%) might be considered promising sources of antiviral natural products, as they have shown EC50 ≤ 100 µg ml(-1). The present screening discloses the high potential of the Bignoniaceae family as source of antiviral agents. SIGNIFICANCE AND IMPACT OF THE STUDY: Active extracts were identified and deserve bioguided studies for the isolation of antiviral compounds and studies on mechanism of action.

Antinociceptive and anti-inflammatory activities of ethanolic extracts of Lychnophora species.

Extracts from Lychnophora species are traditionally used in Brazil as anti-inflammatory, and to treat bruise, pain and rheumatism. The ethanolic extract of aerial parts of five species of Lychnophoras and one specie of Lychnophoriopsis were examined for the antinociceptive (hot-plate and writhing tests) and anti-inflammatory (carrageenan-induced paw oedema test) activity in mice, by oral and topical routes, respectively. In the hot-plate test, the Lychnophora pinaster (0.75 g/kg) and Lychnophora ericoides (1.50 g/kg) extracts significantly increased the time for licking of the paws. The species Lychnophora passerina, Lychnophoriopsis candelabrum and Lychnophora pinaster, using the dose of 0.75 g/kg, and Lychnophora ericoides and Lychnophora trichocarpha in both doses evaluated (0.75 and 1.50 g/kg) significantly reduced the number of writhes induced by acetic acid. The administration of Lychnophora pinaster and Lychnophora trichocarpha ointments, in both concentrations evaluated (5 and 10%, w/w), and Lychnophora passerina and Lychnophoriopsis candelabrum, in the concentration of 10%, significantly reduced the paw oedema measured 3 h after carrageenan administration, suggesting, for the first time, an anti-inflammatory activity upon topical administration of these species. The present work comparatively demonstrated the antinociceptive and anti-inflammatory activities of some Brazilian Lychnophoras.

Xanthine oxidase inhibitory activity of Lychnophora species from Brazil ("Arnica").

Twenty-two extracts from five Lychnophora species and one Lychnophoriopsis species, traditionally used in Brazil as analgesic, anti-inflammatory, and to treat bruise and rheumatism were examined for the inhibition of xanthine oxidase (XO), the enzyme that catalyses the metabolism of hypoxanthine and xanthine into uric acid. Sixteen extracts were tested. All of them were found to have excellent XO inhibitory activity, with inhibitions greater than 38% at 100 microg/mL in the assay mixture. The most active plants examined were Lychnophora trichocarpha, Lychnophora ericoides, Lychnophora staavioides and Lychnophoriopsis candelabrum, with inhibitions of 77%, 78%, 66% and 63% at 100 microg/mL, respectively, and IC(50) values of 6.16, 8.28, 33.97 and 37.70 microg/mL, respectively.

Biological activity of the aqueous extract of Lychnophora pinaster Mart.

Lyophilized aqueous extract (LAE) from Lychnophora pinaster Mart (Asteraceae) aerial parts was evaluated in the search of possible biological activities. LAE exhibited trypanocidal activity (113.62 mg/mL), but could not inhibit 5-lipoxygenase in vitro (17 percent of inhibition). LAE chemical characterization by HPLC with UV-Diode Array Detector showed the presence of caffeic acid, isochlorogenic acid, vitexin, isovitexin and quercetin, in comparison with authentic samples.

Validation of a colorimetric assay for the in vitro screening of inhibitors of angiotensin-converting enzyme (ACE) from plant extracts.

A new method for the in vitro screening of plant extracts with potential angiotensin-converting enzyme (ACE) inhibitory activity is proposed. The method is based on the cleavage of the substrate hippuryl-glycyl-glycine by ACE and subsequent reaction with trinitrobenzenesulfonic acid to form 2,4,6-trinitrophenyl-glycyl-glycine, whose absorbance is determined at 415 nm in a microtitre plate reader. Rabbit lung dehydrated by acetone was employed as an enzyme source. Validation of the method showed satisfactory intra-day (CV = 7.63%) and inter-day precision (CV = 13.61%), recovery (97-102.1%), sensitivity (IC50 = 14.1 nmol/l) and linearity in the range 7.5-120 mmol/l of glycyl-glycine (r2 = 0.9921). Besides, the method showed good correlation with a HPLC assay already established for the screening of ACE inhibitors (r = 0.9935 and 0.9034, respectively, for captopril solutions and for plant extracts). The method involves only inexpensive reagents and apparatus.

Estudo comparativo dos óleos voláteis de algumas espécies de Piperaceae / Comparative study of the essential oils of some species of Piperaceae

O presente trabalho teve por objetivo a análise comparativa dos óleos voláteis de espécies da família Piperaceae. As espécies estudadas foram as seguintes: Piper aduncum, P. amalago, P. arboreum, P. cernuum, P. hispidum, P. regnelii, P. submarginalum, P. vicosanum e Pothomorphe umbellata. A análise dos óleos voláteis foi efetuada pela combinação de técnicas de Cromatografia de Fase Gasosa (CG) e de Cromatografia de Fase Gasosa acoplada a Espectrometria de Massa (CG/EM). A porcentagem de identificação dos constituintes nas análises dos óleos voláteis de Piper aduncum, P. amalago, P. vicosanum, P. submarginalum e Pothomorphe umbellata, atingiu valores superiores a 80 por cento. Houve predominância de compostos de natureza sesquiterpênica nas composições químicas das espécies analisadas, com exceção de P. hispidum, P. submarginalum e P. vicosanum onde a predominância na composição foi de monoterpenos. Pelos dados apresentados observou-se que, dentre os compostos majoritários, os mais freqüentes, nas 9 espécies estudadas, foram beta-pineno (9/9) e espatulenol (9/9), seguidos por E-cariofileno (8/9), óxido de cariofileno (8/9), germacreno D (7/9), alfa-pineno (7/9) e limoneno (6/9). Pela primeira vez se relata a composição dos óleos voláteis de P. submarginalum e P vicosanum.

Essential oils from various vegetal species of the Piperaceae family were analysed by Capillary Gas Chromatography (GC) and Gas Chromatography/Mass Spectrometry (GC/MS). The investigated species were: Piper aduncum, P. amalago, P. arboreum, P. cernuum, P. hispidum, P. regnelii, P. submarginalum, P. vicosanum and Pothomorphe umbellata. Over 80 percent of the essential oils compounds were identified by the qualitative and quantitative analysis. The analysed species showed the predominance of sesquiterpenoids, except for P. hispidum, P. submarginalum and P. vicosanum in which the predominant compounds were monoterpenoids. For the 9 studied species, the major compounds found in the oils were beta-pinene (9/9) and spathulenol (9/9), followed by the E-caryophyllene (8/9), caryophyllene oxide (8/9), germacrene D (7/9), alpha-pinene (7/9) and limonene (6/9).

Estudo fitoquímico de Erythraea centaurium, Jacaranda caroba, Remijia ferruginea e Solanum paniculatum visando identificar marcadores químicos para o fitoterápico Ierobina®

O fitoterápico Ierobina®, comercializado no mercado nacional há 67 anos, tem uso indicado para o tratamento de dispepsias. O fitoterápico é constituído pelos extratos fluidos de três espécies nativas (Solanum paniculatum, Remijia ferruginea e Jacaranda caroba) e uma exótica (Erythraea centaurium). O presente estudo descreve a identificação de marcadores químicos para os extratos das espécies constituintes da Ierobina® e a obtenção de perfis cromatográficos de referência para os mesmos, os quais contribuirão para o estabelecimento de protocolos para o controle de qualidade do fitoterápico.

Isolamento biomonitorado de uma substância tripanossomicida de Arrabidaea triplinervia (Bignoniaceae), o ácicIo ursólico

Como estratégia para a busca de novos agentes tripanossomicidas, de potencial utilização no controle da transmissão transfusional da doença de Chagas, realizou‑se uma triagem dos extratos etanólicos provenientes de espécies vegetais da família Bignoniaceae. Foram avaliadas diferentes metodologias de fracionamento dos extratos etanólicos, tais como, partição entre solventes imisciveis, cromatografias em coluna de poliamida e de gel de sílica, com monitoramento por testes in vitro contra formas tripomastigotas do Trypanosoma cruzi. Os fracionamentos em coluna de gel de sílica do extrato etanólico das folhas de Arrabidaea triplinervia e, posteriormente, da fração eluída desta coluna com diclorometano‑acetato de etila (1:1) resultaram no isolamento de uma substância tripanossomicida, o ácido ursólico (0,76 por cento).

Age differences and changes in midline-crossing inhibition in the lower extremities.

BACKGROUND: The effects of crossing the body midline on the lower extremities of individuals 65 years of age and older was investigated. METHODS: The subjects were 10 individuals 65-79 years of age, 10 individuals 80 years and older, and 10 individuals 20-35 years. The total testing phase consisted of 2 sets of 30 trials per leg on 2 days. The subjects performed trials that involved movements ipsilaterally, contralaterally, and directly in front of the anterior superior iliac crest of the leg being tested. Reaction time and movement time scores were recorded. RESULTS: Individuals 65 years of age and older were found to exhibit slower reaction times to movements in a contralateral direction when compared with movements made in the ipsilateral direction whereas individuals 80 years of age and older were also found to exhibit slower reaction times to movements in a contralateral direction when compared with movements made in the midline and ipsilateral direction. CONCLUSIONS: These findings indicate that the effects of midline-crossing inhibition on the lower extremities reemerge in individuals 65 years and older, whereas in early development this effect disappears by 8 or 9 years of age.

Second-look arthroscopy with removal of bioabsorbable tacks.

Eleven years after tearing her anterior cruciate ligament (ACL) (not reconstructed), a 36-year-old dancer reinjured her knee and required arthroscopic ACL reconstruction. At arthroscopy, the medial meniscus had a bucket-handle tear that was repaired by using three bioabsorbable tacks. The ACL was then repaired in the usual manner. Because of persistent posterior knee pain throughout her rehabilitation, we performed "second-look" arthroscopy 14 weeks after reconstruction. The meniscus had healed and was stable; however, tack motion was evident and the tacks were easily removed. Inspection of the tacks showed that the barbs had been resorbed. The patient recovered uneventfully, and pain-free flexion 28 days after surgery was 0 degrees-136 degrees. We believe this to be the first reported case demonstrating the early stages of tack degradation in meniscal repair.

Screening Brazilian plant species for in vitro inhibition of 5-lipoxygenase.

Plants from the Brazilian flora were evaluated for the inhibition of 5-lipoxygenase. The species were selected based on their traditional use and on a chemosystematic approach. In total, 19 species belonging to 13 families have been investigated. Hedychium coronarium J. Koenig (Zingiberaceae), Xylopia frutescens Aubl. (Annonaceae) and Hymenaea courbaril L. (Leguminosae) presented a high 5-lipoxygenase inhibitory activity. Some hypothesis about the nature of the active compounds are discussed, based on reports of the chemical constitution of these species or other species from the same botanical family.

Screening the Brazilian flora for antihypertensive plant species for in vitro angiotensin-I-converting enzyme inhibiting activity.

The evaluation of several antihypertensive activity of Brazilian plant species was performed using in vitro inhibition of the angiotensin I-converting enzyme (ACE). Nineteen species belonging to 13 families were investigated. Plants were selected based on their use as diuretics and on a chemosystematic consideration. Extracts of the following species presented the highest ACE inhibition rate, at concentrations of 0.33 mg/ml: Ouratea semiserrata (Mart. & Nees) Engl. stems (68%), Cuphea cartagenesis (Jacq.) Macbride leaves (50%) and Mansoa hirsuta DC. leaves (54%). Some hypotheses about the nature of the compounds that may be responsible for the activity of these species are discussed in the paper.