Research progress on chemical constituents and pharmacological activities of Viola plants / 中国中药杂志

There are 500 species of Viola(Violaceae) worldwide, among which 111 species are widely distributed in China and have a long medicinal history and wide varieties. According to the authors' statistics, a total of 410 compounds have been isolated and identified from plants of this genus, including flavonoids, terpenoids, phenylpropanoids, organic acids, nitrogenous compounds, sterols, saccharides and their derivatives, volatile oils and cyclotides. The medicinal materials from these plants boast anti-microbial, anti-viral, anti-oxidant and anti-tumor activities. This study systematically reviewed the chemical constituents and pharmacological activities of Viola plants to provide a basis for further research and clinical application.

Five new terpenoids from Viburnum odoratissimum var. sessiliflorum / 中国天然药物

Five new terpenoids, including two vibsane-type diterpenoids (1, 2) and three iridoid allosides (3-5), together with eight known ones, were isolated from the leaves and twigs of Viburnum odoratissimum var.sessiliflorum. Their planar structures and relative configurations were determined by spectroscopic methods, especially 2D NMR techniques. The sugar moieties of the iridoids were confirmed as β-D-allose by GC analysis after acid hydrolysis and acetylation. The absolute configurations of neovibsanin Q (1) and dehydrovibsanol B (2) were determined by quantum chemical calculation of their theoretical electronic circular dichroism (ECD) spectra and Rh2(OCOCF3)4-induced ECD analysis. The anti-inflammatory activities of compounds 1, 3, 4, and 5 were evaluated using an LPS-induced RAW264.7 cell model. Compounds 3suppressed the release of NO in a dose-dependent manner, with an IC50 value of 55.64 μmol·L-1. The cytotoxicities of compounds 1-5 on HCT-116 cells were assessed and the results showed that compounds 2 and 3 exhibited moderate inhibitory activities with IC50 values of 13.8 and 12.3 μmol·L-1, respectively.

Research progress of Curcuma kwangsiensis root tubers and analysis of liver protection and anti-tumor mechanisms based on Q-markers / 中国中药杂志

Curcuma kwangsiensis root tuber is a widely used genuine medicinal material in Guangxi, with the main active components of terpenoids and curcumins. It has the effects of promoting blood circulation to relieve pain, moving Qi to relieve depression, clearing heart and cooling blood, promoting gallbladder function and anti-icterus. Modern research has proved its functions in liver protection, anti-tumor, anti-oxidation, blood lipid reduction and immunosuppression. Considering the research progress of C. kwangsiensis root tubers and the core concept of quality marker(Q-marker), we predicted the Q-markers of C. kwangsiensis root tubers from plant phylogeny, chemical component specificity, traditional pharmacodynamic properties, new pharmacodynamic uses, chemical component measurability, processing methods, compatibility, and components migrating to blood. Curcumin, curcumol, curcumadiol, curcumenol, curdione, germacrone, and β-elemene may be the possible Q-markers. Based on the predicted Q-markers, the mechanisms of the liver-protecting and anti-tumor activities of C. kwangsiensis root tubers were analyzed. AKT1, IL6, EGFR, and STAT3 were identified as the key targets, and neuroactive ligand-receptor interaction signaling pathway, nitrogen metabolism pathway, cancer pathway, and hepatitis B pathway were the major involved pathways. This review provides a basis for the quality evaluation and product development of C. kwangsiensis root tubers and gives insights into the research on Chinese medicinal materials.

Antibiofilm efficacy of tea tree oil and of its main component terpinen-4-ol against Candida albicans

Abstract Candida infection is an important cause of morbidity and mortality in immunocompromised patients. The increase in its incidence has been associated with resistance to antimicrobial therapy and biofilm formation. The aim of this study was to evaluate the efficacy of tea tree oil (TTO) and its main component - terpinen-4-ol - against resistant Candida albicans strains (genotypes A and B) identified by molecular typing and against C. albicans ATCC 90028 and SC 5314 reference strains in planktonic and biofilm cultures. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity. Results were obtained from analysis of the biofilm using the cell proliferation assay 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and confocal laser scanning microscopy (CLSM). Terpinen-4-ol and TTO inhibited C. albicans growth. CLSM confirmed that 17.92 mg/mL of TTO and 8.86 mg/mL of terpinen-4-ol applied for 60 s (rinse simulation) interfered with biofilm formation. Hence, this in vitro study revealed that natural substances such as TTO and terpinen-4-ol present promising results for the treatment of oral candidiasis.

Effect of thymol and environmental factors on growth and biofilm formation by Listeria monocytogenes / Efecto del timol y factores ambientales sobre el crecimiento y formación de biopelícula por Listeria monocytogenes

Thymol (2-isopropyl-5-methylphenol) is an aromatic monoterpene found in essential oils extracted from plants belonging to the Lamiaceae family, such as Thymus, Ocimum, Origanum, Satureja, Thymbra and Monarda genera. Growth and biofilm formation by Listeria monocytogenes CLIP 74902 were evaluate using three carbon sources in the presence of thymol. Specific growth rate (h-1) values at 37o with glucose, trehalose and cellobiose with the addition of thymol (µg/mL) 0 (control) and 750, were respectively: 0.22, 0.07; 0.14, 0.04; 0.11, 0.04. Lag periods obtained under the same conditions were (h): 8.19, 13.2; 22.5, 27.5; 23.1, 28.1. A marked antibiofilm activity was observed against the exposure with 750 µg/mL of thymol, showing a high percentage of inhibition: glucose (99 %), trehalose (97 %) and cellobiose (98%), compared to the control. The results suggest that thymol could be used to inhibit the growth and production of biofilms by L. monocytogenes in the food industry.

Timol (2-isopropil-5-metilfenol) es un monoterpeno aromático presente en los aceites esenciales extraídos de plantas pertenecientes a la familia Lamiaceae, como los géneros Thymus, Ocimum, Origanum, Satureja, Thymbra y Monarda. El crecimiento y formación de biopelícula por Listeria monocytogenes CLIP 74902 fueron evaluados utilizando tres fuentes de carbono en presencia de timol. La velocidad específica de crecimiento (h-1) a 37o con glucosa, trehalosa y celobiosa con la adición de timol (µg/mL) 0 (control) y 750, fueron respectivamente: 0.22, 0.07; 0.14, 0.04, 0.11, 0,04. Los períodos lag obtenidos en las mismas condiciones fueron (h): 8.19, 13.2; 22.5, 27.5; 23.1, 28.1. Una marcada actividad antibiofilm fue obtenida con 750 µg/mL de timol, mostrando un alto porcentaje de inhibición con glucosa (99%), trehalosa (97%) y celobiosa (98%), respecto al control. Los resultados sugieren que timol podría ser usado para inhibir el crecimiento y producción de biopelículas por L. monocytogenes en la industria alimentaria.

Evaluation in vitro of proliferative activity of epithelial cells by flavonoid 3-O-methylgalangine and terpenenic derivative Filifolinone / Evaluación in vitro de la actividad proliferative de células epiteliales del flavonoide 3-O-metilgalangina y el derivado terpénico Filifolinona

The skin is the largest organ of the human body and its main function is to protect it from the external environment. It is exposed to injuries that require a rapid healing process to recover its functionality. Microorganisms inhabit the skin, which makes up the normal microbial flora, but in situations of injury they can cause infections that slow down the regeneration process. Therefore, there is a great interest in the development of alternative methods to accelerate the regeneration process and prevent infections. In this work, the efficacy of flavonoid 3-O-methylgalangine and the terpenic derivative Filifolinone and its mixtures, isolated from plants of the genus Heliotropium, on the stimulation of cell proliferation was evaluated. The results showed that the mixtures stimulated proliferation and migration in MA104 cells mainly due to the presence of Filifolinone, that together with the known antibacterial activity of 3-O-methylgalangine, opens new alternatives for the use of natural compounds in healing processes.

La piel es el órgano más grande del cuerpo humano y su función principal es protegerla del entorno externo. Está expuesta a lesiones que requieren un proceso de curación rápido para recuperar su funcionalidad. Los microorganismos que habitan en la piel, constituyen la flora microbiana normal, pero en situaciones de lesión pueden causar infecciones que retardan el proceso de regeneración. Por lo tanto, existe un gran interés en el desarrollo de métodos alternativos para acelerar el proceso de regeneración y prevenir infecciones. En este trabajo, se evaluó la eficacia del flavonoide 3-O-metilgalangina y el derivado terpénico Filifolinona y sus mezclas, aisladas de plantas del género Heliotropium, en la estimulación de la proliferación celular. Los resultados mostraron que las mezclas estimularon la proliferación y la migración en las células MA104 debido principalmente a la presencia de Filifolinona, que junto con la actividad antibacteriana conocida de la 3-O-metilgalangina, abre nuevas alternativas para el uso de compuestos naturales en los procesos de curación.

Interactions between Terpinen-4-ol and Nystatin on biofilm of Candida albicans and Candida tropicalis

Abstract The aim of this study was to evaluate the antifungal activity of Terpinen-4-ol associated with nystatin, on single and mixed species biofilms formed by Candida albicans and Candida tropicalis, as well as the effect of terpinen-4-ol on adhesion in oral cells and the enzymatic activity. The minimum inhibitory concentrations and minimum fungicide concentrations of terpinen-4-ol and nystatin on Candida albicans and Candida tropicalis were determined using the microdilution broth method, along with their synergistic activity ("checkerboard" method). Single and mixed species biofilms were prepared using the static microtiter plate model and quantified by colony forming units (CFU/mL). The effect of Terpinen-4-ol in adhesion of Candida albicans and Candida tropicalis in coculture with oral keratinocytes (NOK Si) was evaluated, as well as the enzymatic activity by measuring the size of the precipitation zone, after the growth agar to phospholipase, protease and hemolysin. Terpinen-4-ol (4.53 mg mL-1) and nystatin (0.008 mg mL-1) were able to inhibit biofilms growth, and a synergistic antifungal effect was showed with the drug association, reducing the inhibitory concentration of nystatin up to 8 times in single biofilm of Candida albicans, and 2 times in mixed species biofilm. A small decrease in the adhesion of Candida tropicalis in NOK Si cells was showed after treatment with terpinen-4-ol, and nystatin had a greater effect for both species. For enzymatic activity, the drugs showed no action. The effect potentiated by the combination of terpinen-4-ol and nystatin and the reduction of adhesion provide evidence of its potential as an anti-fungal agent.

Resumo O objetivo desse estudo foi avaliar a atividade antifúngica do Terpinen4-ol associado à nistatina em biofilmes simples e misto, formados por Candida albicans e Candida tropicalis, bem como o efeito do terpinen-4-ol na adesão em células orais e atividade enzimática. As concentrações inibitórias mínimas e as concentrações fungicidas mínimas do terpinen-4-ol e da nistatina em Candida albicans e Candida tropicalis foram determinadas pelo método de microdiluição em caldo, juntamente com a atividade sinérgica (método do tabuleiro de "xadrez"). Biofilmes simples e misto foram preparados usando o modelo de placa de microtitulação estática e quantificados por unidades formadoras de colônias (CFU/mL). O efeito do Terpinen-4-ol na adesão de Candida albicans e Candida tropicalis em co-cultura com queratinócitos orais (NOK Si) foi avaliado, bem como a atividade enzimática, medindo o tamanho da zona de precipitação, após o crescimento em ágar fosfolipase, protease e hemolisina. O terpinen-4-ol (4.53 mg mL-1) e a nistatina (0,008 mg mL-1) conseguiram inibir o crescimento de biofilmes e um efeito antifúngico sinérgico foi demonstrado com a associação de fármaco, reduzindo a concentração inibidora de nistatina até 8 vezes em biofilme simpes de Candida albicans e 2 vezes em biofilme misto. Uma pequena diminuição na adesão de Candida tropicalis em células NOK Si foi mostrada após o tratamento com terpinen-4-ol e a nistatina teve um efeito maior para ambas as espécies. Para a atividade enzimática, as drogas não apresentaram ação. O efeito potencializado pela combinação de terpinen-4-ol e nistatina e a redução de adesão evidenciam seu potencial como agente anti-fúngico.

Terpinen-4-ol, tyrosol, and ß-lapachone as potential antifungals against dimorphic fungi

Abstract This study aimed to evaluate the in vitro antifungal activity of terpinen-4-ol, tyrosol, and β-lapachone against strains of Coccidioides posadasii in filamentous phase (n = 22) and Histoplasma capsulatum in both filamentous (n = 40) and yeast phases (n = 13), using the broth dilution methods as described by the Clinical and Laboratory Standards Institute, to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of these compounds. The mechanisms of action of these compounds were also investigated by analyzing their effect on cell membrane permeability and ergosterol synthesis. The MIC and MFCf these compounds against C. posadasii, mycelial H. capsulatum, and yeast-like H. capsulatum, were in the following ranges: 350-5720 µg/mL, 20-2860 µg/mL, and 40-1420 µg/mL, respectively for terpinen-4-ol; 250-4000 µg/mL, 30-2000 µg/mL, and 10-1000 µg/mL, respectively, for tyrosol; and 0.48-7.8 µg/mL, 0.25-16 µg/mL, and 0.125-4 µg/mL, respectively for β-lapachone. These compounds showed a decrease in MIC when the samples were subjected to osmotic stress, suggesting that the compounds acted on the fungal membrane. All the compounds were able to reduce the ergosterol content of the fungal strains. Finally, tyrosol was able to cause a leakage of intracellular molecules.

Insecticidal activity of neem oil against Gyropsylla spegazziniana (Hemiptera: Psyllidae) nymphs on Paraguay tea seedlings / Atividade inseticida de óleo de nim contra ninfas de Gyropsylla spegazziniana (Hemiptera: Psyllidae) em mudas de erva-mate

Abstract Gyropsylla spegazziniana (Paraguay tea ampul) is one of the most important pests of Paraguay tea plants, and prohibition of synthetic insecticide use for control of this pest has led to the search for alternative methods. This laboratory study aimed to compare different control strategies for G. spegazziniana, utilizing a commercial neem seed oil product. Paraguay tea seedlings were treated with neem oil solution both pre- and post-infestation with 5th instar nymphs. The systemic action of neem oil was also evaluated by treating plant soil with the neem oil solution, followed by transfer of the insects to plants 24 h post-treatment. Spray treatments were effective against the pest, especially post-infestation (80% mortality), demonstrating the potential of neem oil for control of the Paraguay tea ampul. No significant effects were observed with respect to systemic activity.

Resumo Gyropsylla spegazziniana (ampola da erva-mate) é uma das principais pragas da erva-mate, tanto em áreas de plantio como em viveiros. A proibição do uso de inseticidas para o controle de pragas da cultura tem levado à busca de alternativas. Este trabalho visou avaliar comparativamente, em laboratório, estratégias de aplicação de um produto comercial à base de óleo de nim sobre G. spegazziniana, em solução a 5%. Os testes foram realizados em mudas de erva-mate, tratadas pré e pós-infestação com ninfas de 5º instar do inseto, para avaliação de contato. Também, avaliou-se a ação sistêmica do óleo de nim, aplicando-se a solução do produto no solo da muda de erva-mate, seguido da transferência dos insetos após 24 h. Verificou-se que os tratamentos via pulverização foram eficientes contra a praga, principalmente na aplicação pós-infestação (80% de mortalidade), comprovando o potencial inseticida do óleo de nim contra a ampola-da-erva-mate em mudas. Entretanto, em relação à atividade sistêmica, não foram observados efeitos consideráveis sobre a praga.

Citotoxicidad/capacidad antiradicalaria in-vitro y caracterización estructural por GC-MS/1H-13C-RMN de los aceites esenciales de hojas de árboles joven/adulto de annona purpurea Moc. & Sessé ex Dunal de Repelón (Atlántico, Colombia) / [in-vitro Cytotoxicity/Anti-Radical Capability and structural characterization by GC-MS/1H13C-NMR of theleaves essential oils of young/old trees of Annona purpurea Moc. & Sessé ex Dunal from Repelón (Atlántico,Colombia]

Annona purpurea es una planta empleada en etnomedicina, en la región norte de Colombia, que no tiene reportes científicos relacionados con la composición de su AE y sus actividades biológicas. En este trabajo, los AE de hojas de árboles joven/adulto se caracterizaron por GC-MS y RMN y se evaluaron sus propiedades antiradicalarias/citotóxicas. Los AE de hojas de árboles joven/adulto estuvieron representados por beta-eudesmol (68.9 por ciento) y alfa-eudesmol (16.8 por ciento), y germacreno D (55.6 por ciento) y biciclogermacreno (20.3 por ciento), respectivamente. El análisis por RMN mostró las señales distintivas de los constituyentes mayoritarios identificados. Los valores de TAA (mmol Trolox®/kg SE) obtenidos por los AE de árboles joven/adulto fueron 165 +/- 8 y 602 +/- 38. Los AE de árboles joven/adulto evaluados sobre linfocitos humanos fueron “moderadamente tóxicos” con valores de CL50 (ug/mL) de 145.5 +/- 0.7 y 346 +/- 8. Finalmente, la citotoxicidad en eritrocitos humanos reveló que el AE de árbol adulto no fue hemolítico (CL50 > 1000 μg/mL, 4.3 +/- 0.6 por ciento); mientras que, el AE de árbol joven fue hemolítico (CL50 490 +/- 48 ug/mL).

Annona purpurea is a plant used in ethnomedicine in the northern region of Colombia, which has no scientific reports on the composition of their essential oil (EO) and biological activities. In this work, the leaves EO of young/old trees were characterized by GC- MS and NMR, and their antiradical/cytotoxic properties were evaluated. beta-Eudesmol (68.9 percent) and alpha-eudesmol (16.8 percent), and germacrene D (55.6 percent) and bicyclogermacrene (20.3 percent), were the representative compounds of the leaves EO of young/old trees, respectively. The NMR analysis showed the distinctive signals of the main constituents identified. The TAA values (mmol Trolox®/kg ES) obtained from the EO of young/old trees were 165 +/- 8 and 602 +/- 38. The EO of young/old trees evaluated on human lymphocytes were “moderately toxic” with LC50 (μg/mL) of 145.5 +/- 0.7 and 346 +/- 8. Finally, the cytotoxicity in human erythrocytes revealed that the old tree EO was not haemolytic (LC50 > 1000 ug/mL, 4.3 +/- 0.6 percent); while the young tree EO was hemolytic (LC50 490 +/- 48 ug/mL).

Association of terpinolene and diclofenac presents antinociceptive and anti-inflammatory synergistic effects in a model of chronic inflammation

Pharmacological treatment of inflammatory pain is usually done by administration of non-steroidal anti-inflammatory drugs (NSAIDs). These drugs present high efficacy, although side effects are common, especially gastrointestinal lesions. One of the pharmacological strategies to minimize such effects is the combination of drugs and natural products with synergistic analgesic effect. The monoterpene terpinolene (TPL) is a chemical constituent of essential oils present in many plant species, which have pharmacological activities, such as analgesic and anti-inflammatory. The association of ineffective doses of TPL and diclofenac (DCF) (3.125 and 1.25 mg/kg po, respectively) presented antinociceptive and anti-inflammatory effects in the acute (0, 1, 2, 3, 4, 5 and 6 h, after treatment) and chronic (10 days) inflammatory hyperalgesia induced by Freund's complete adjuvant (CFA) in the right hind paw of female Wistar rats (170-230 g, n=6-8). The mechanical hyperalgesia was assessed by the Randall Selitto paw pressure test, which determines the paw withdrawal thresholds. The development of edema was quantified by measuring the volume of the hind paw by plethismography. The TPL/DCF association reduced neutrophils, macrophages and lymphocytes in the histological analysis of the paw, following a standard staining protocol with hematoxylin and eosin and the counts were performed with the aid of optical microscopy after chronic oral administration of these drugs. Moreover, the TPL/DCF association did not induce macroscopic gastric lesions. A possible mechanism of action of the analgesic effect is the involvement of 5-HT2A serotonin receptors, because ketanserin completely reversed the antinociceptive effect of the TPL/DCF association. These results suggest that the TPL/DCF association had a synergistic anti-inflammatory and analgesic effect without causing apparent gastric injury, and that the serotonergic system may be involved in the antinociceptive effect of this association.

Effect of myrrh and thyme on Trichinella spiralisenteral and parenteral phases with inducible nitric oxide expression in mice

Trichinellosis is a serious disease with no satisfactory treatment. We aimed to assess the effect of myrrh (Commiphora molmol) and, for the first time, thyme (Thymus vulgaris L.) against enteral and encysted (parenteral) phases of Trichinella spiralis in mice compared with albendazole, and detect their effect on inducible nitric oxide synthase (iNOS) expression. Oral administration of 500 mg/kg of myrrh and thyme led to adult reduction (90.9%, 79.4%), while 1,000 mg/kg led to larvae reduction (79.6%, 71.3%), respectively. Administration of 50 mg/kg of albendazole resulted in adult and larvae reduction (94.2%, 90.9%). Positive immunostaining of inflammatory cells infiltrating intestinal mucosa and submucosa of all treated groups was detected. Myrrh-treated mice showed the highest iNOS expression followed by albendazole, then thyme. On the other hand, both myrrh and thyme-treated groups showed stronger iNOS expression of inflammatory cells infiltrating and surrounding encapsulated T. spiralis larvae than albendazole treated group. In conclusion, myrrh and thyme extracts are highly effective against both phases of T. spiralis and showed strong iNOS expressions, especially myrrh which could be a promising alternative drug. This experiment provides a basis for further exploration of this plant by isolation and retesting the active principles of both extracts against different stages of T. spiralis.

effect of neem [Azadirachta indica A. Juss] thevetia [thevetia peruviana [Pers.] K. Schum.] extracts inhibition of fungi growth causing cotton root-rot disease

The present showed the effect of Neem and Thevetia extract on growth of R. solani. The Stsdy Neem and Thevetia oil seed inhibited the growth of R. solani percentage of inhibition. Data indicated that the inhibition percentage of oil seed for the Neem was 79.3% and for Thevetia it was 75.2%. Ethanolic extract of leaves of Neem and Thevetia showed higher activity on the growth of R. solani at higher concentration. Applying ethanolic extract of the leaves of the Neem and Thevetia, revealed that the inhibition of the extract of leaves on the Neem was markedly higher than of Thevetia [68.9% versus 48.9%]

Potencial inseticida de óleos de origem vegetal sobre Grapholita molesta (Busck) (Lepidoptera: Tortricidae) / Insecticidal potential of vegetable oil extracts on Grapholita molesta (Busck) (Lepidoptera: Tortricidae)

Em decorrência do estudo das interações químicas entre insetos e herbívoros, e do avanço da pesquisa fitoquímica, é conhecida ampla variedade de produtos naturais com potencial inseticida. Grapholita molesta (Busck) (Lepidoptera: Tortricidae), mariposa-oriental, é uma das principais pragas do pessegueiro, danificando brotações e frutos. O objetivo do presente estudo foi verificar a mortalidade, atratividade e deterrência de óleos vegetais em G. molesta. Ovos e pupas de insetos provindos de criação artificial foram imersos nos óleos essenciais de Elionurus muticus (Spreng.) Kuntze e Cymbopogon winterianus Jowitt ex Bor nas concentrações de 0,25%, 0,5%, 1% e 0,5%, 1%, 5%. Os testes de atratividade foram realizados em olfatômetro do tipo "Y". A deterrência à oviposição foi testada em gaiolas sem escolha e de dupla escolha. O óleo de E. muticus aplicado em ovos de G. molesta nas três concentrações causou mortalidades maiores que 30%, diferindo significativamente dos controles (p<0,05). A mortalidade causada pelo óleo de citronela foi em torno de 70%, significativamente superior ao controle (p<0,01). O óleo de citronela aplicado a 1% em pupas causou mortalidade de 99,8%. No teste com o olfatômetro, não houve atratividade dos adultos para nenhum dos óleos. A média de ovos no substrato com o tratamento (0,33±0,33), foi significativamente menor que no controle (7,3±0,88) (p<0,01), indicando deterrência à oviposição.

As a result of the study on the chemical interactions between insects and herbivores and the advancement of phytochemical research studies, a variety of natural products with great insecticidal potential are described in the literature. The Grapholita molesta (Busck) (Lepidoptera: Tortricidae), commonly known as Oriental Fruit Moth, is a major peach pest damaging shoots and fruits. The purpose of this study was to assess the mortality, attractiveness and deterrence of vegetable oils in G. molesta. Insect eggs and pupae artificially created were immersed in the essential oils of Elionurus muticus (Spreng.) Kuntze and/or Cymbopogon winterianus Jowitt ex Bor at the concentrations of 0.25%, 0.5%, 1% and 0.5%, 1%, 5%, respectively. The attractiveness tests were performed in a Y-Tube olfactometer. Oviposition deterrence was tested in no-choice and dual-choice cages. Oil of E. muticus applied in eggs of G. molesta at three concentrations caused mortality above 30%, which differs significantly from controls (p<0.05). Mortality caused by citronella oil was approximately 70%, significantly higher than control (p<0.01). The application of citronella oil at 1% in pupae caused mortality of 99.8%. There was no attractiveness of adults in the olfactometer test for any of the oils. Citronella oil presented oviposition deterrence, as the mean number of eggs in the substrate with the treatment (0.33 ± 0.33) is significantly lower than in the control (7.3 ± 0.88) (p<0.01).

Candida albicans biofilm inhibition by synergistic action of terpenes and fluconazole.

The current treatment options for Candida albicans biofilm-device related infections are very scarce due to their intrinsic increased tolerance to antimycotics. The aim of this work was to study synergistic action of terpenes (eugenol, menthol and thymol) with fluconazole (FLA) on C. albicans biofilm inhibition. The minimum inhibitory concentration (MIC) assayed using CLSI M27-A3 broth micro-dilution method showed antifungal activity against C. albicans MTCC 227 at a concentration of 0.12 % (v/v) for both thymol and eugenol as compared to 0.25 % (v/v) for menthol. FLA was taken as positive control. The effect of these terpenes on metabolic activity of preformed C. albicans biofilm cells was evaluated using 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assay in 96-well polystyrene microtiter plate. Thymol and eugenol were more effective at lower concentrations of ≥ 1.0 % (v/v) than menthol. Synergistic studies using checkerboard micro-dilution assay showed fractional inhibitory concentration index (Σ FIC=0.31) between thymol/FLA followed by eugenol/FLA (Σ FIC=0.37) and menthol/FLA (Σ FIC<0.5) against pre-formed C. albicans biofilms. Thymol with fluconazole showed highest synergy in reduction of biofilm formation than eugenol and menthol which was not observed when their activities were observed independently. Adherence assay showed 30% viability of C. albicans cells after 2 h of treatment with 0.05 % (v/v) thymol/FLA. Effect of thymol/FLA on C. albicans adhesion visualized by SEM micrographs showed disruption in number of candidal cells and alteration in structural design of C. albicans. Thus, the study demonstrated synergistic effect of terpenes with fluconazole on C. albicans biofilm, which could be future medications for biofilm infections.

Sedative and anesthetic activities of the essential oils of Hyptis mutabilis (Rich.) Briq. and their isolated components in silver catfish (Rhamdia quelen)

This study evaluated the sedative and anesthetic effects of the essential oils (EO) of Hyptis mutabilis (Rich.) Briq. and their isolated components on silver catfish (Rhamdia quelen). Quantitative chemical differences between the EOs obtained from leaves and inflorescences were verified, and a new chemotype rich in globulol was described. Although there were no significant differences in the time of induction for sedation and anesthesia between the EOs, only the leaf EO at 344 mg/L anesthetized all fish without side effects. Fractionation of the leaf EO was carried out by column chromatography. The isolated compounds [(+)-1-terpinen-4-ol and (-)-globulol] showed different activity from that detected for the leaf EO in proportional concentrations and similar sedation to a eugenol control at 10 mg/L. However, fish exposed to 1-terpinen-4-ol (3 and 10 mg/L) did not remain sedated for 30 min. Anesthesia was obtained with 83-190 mg/L globulol, but animals showed loss of mucus during induction and mortality at these concentrations. Synergism of the depressor effects was detected with the association of globulol and benzodiazepine (BDZ), compared with either drug alone. Fish exposed to BDZ or globulol+BDZ association showed faster recovery from anesthesia in water containing flumazenil, but the same did not occur with globulol. In conclusion, the use of globulol in aquaculture procedures should be considered only at sedative concentrations of 10 and 20 mg/L, and its mechanism of action seems not to involve the GABAA-BDZ system.

In vitro evaluation of new terpenoid derivatives against Leishmania infantum and Leishmania braziliensis

The activity of five (1-5) abietane phenol derivatives against Leishmania infantum and Leishmania braziliensis was studied using promastigotes and axenic and intracellular amastigotes. Infectivity and cytotoxicity tests were performed with J774.2 macrophage cells using Glucantime as a reference drug. The mechanisms of action were analysed by performing metabolite excretion and transmission electron microscopy ultrastructural studies. Compounds 1-5 were more active and less toxic than Glucantime. The infection rates and mean number of parasites per cell observed in amastigote experiments showed that derivatives 2, 4 and 5 were the most effective against both L. infantum and L. braziliensis. The ultrastructural changes observed in the treated promastigote forms confirmed that the greatest cell damage was caused by the most active compound (4). Only compound 5 caused changes in the nature and amounts of catabolites excreted by the parasites, as measured by ¹H nuclear magnetic resonance spectroscopy. All of the assayed compounds were active against the two Leishmania species in vitro and were less toxic in mammalian cells than the reference drug.

Revisión bibliográfica sobre la composición química y actividades farmacológicas del género Vismia (Guttiferae) / A review on the chemical composition and pharmacological activities of Vismia genus (Guttiferae)

Vismia genus is an important source of natural medicinal products, thus, information collected in this review is an attempt to cover the most recent developments in the ethnopharmacology, pharmacology and phytochemistry of this genus. Anthraquinones and other quinonoid derivates, terpenoids and volatile constituents have been reported as the major constituents isolated from different Vismia species. On the other hand, pharmacological studies carried out to date have revealed the variety of anti-plasmodium, antioxidant, antimicrobial and antifungical properties of extracts and pure isolated compounds of the different species tested. The information summarized in this paper intends to serve as a reference tool to practitioners in the fields of etnopharmacology and chemistry of natural products.

El género Vismia es una fuente importante de productos naturales medicinales, es por esto que la información reunida en la presente revisión cubre los estudios más recientes en la etnofarmacología, farmacología y fitoquímica de este género. Antraquinonas y otros derivados quinoides, terpenos y constituyentes volátiles han sido reportados como los compuestos mayormente aislados de las diferentes especies de Vismia. Por otro lado, los estudios farmacológicos realizados hasta los momentos muestran las diversas propiedades antiplasmodicas, antioxidantes, antimicrobianas y antifúngicas que presentan tanto los extractos como los compuestos puros aislados de las diferentes especies ensayadas. La información resumida en este documento intenta servir de material de apoyo para investigadores en los campos de la etnofarmacologia y la química de productos naturales.

Bait formulations of molluscicides and their effects on biochemical changes in the ovotestis of snail Lymnaea acuminata (Mollusca; Gastropoda:Lymnaeidae) / Formulações de iscas de moluscicidas e seus efeitos sobre as alterações bioquímicas no ovoteste do caramujo Lymnaea acuminata (Mollusca;Gastropoda:Lymnaeidae)

The effect of sub-lethal feeding of bait formulations containing molluscicidal component of Ferula asafoetida (ferulic acid, umbelliferone), Syzygium aromaticum (eugenol) and Carum carvi (limonene) on biochemical changes in the ovotestis of snail Lymnaea acuminata were studied. Bait formulations feeding to L. acuminata were studied in clear glass aquaria having diameter of 30 cm. Baits were prepared from different binary combinations of attractant amino acid (valine, aspartic acid, lysine and alanine 10 mM) in 100 mL of 2 percent agar solution + sub-lethal (20 percent and 60 percent of 24h LC50) doses of different molluscicides (ferulic acid, umbelliferone, eugenol and limonene). These baits caused maximum significant reduction in free amino acid, protein, DNA, RNA levels i.e. 41.37, 23.56, 48.36 and 14.29 percent of control in the ovotestis of the snail, respectively. Discontinuation of feeding after treatment of 60 percent of 96h LC50 of molluscicide containing bait for next 72h caused a significant recovery in free amino acid, protein, DNA and RNA levels in the ovotestis of L. acuminata.

Foi estudado o efeito subletal das iscas usadas para alimentação contendo componentes moluscicidas de Ferula asafoetida (ácido ferúlico, umbeliferone), Syzygium aromaticum (eugenol) e Carum carvi (limonene) nas alterações bioquímicas do ovoteste do caramujo Lymnaea acuminata. A formulação das iscas usadas para alimentar L. acuminata foi estudada em aquários de vidros transparentes de diâmetro de 30 cm. As iscas foram preparadas por combinações diferentes binárias de aminoácidos (valina, ácido aspártico, lisina e alanina 10 mM) em 100 mL de solução de agar a 2 por cento + doses subletais (20 por cento e 60 por cento durante 24 horas LC50) de diferentes moluscicidas (ácido ferúlico, umbeliferone, eugenol e limonene). Estas iscas causaram redução significante máxima em aminoácidos livres, proteínas, níveis de DNA e RNA isto é 41,37 por cento, 23,56 por cento, 48,36 por cento e 14,29 por cento de controle no ovoteste do caramujo, respectivamente. Discontinuação da alimentação depois do tratamento de 60 por cento de 96 horas de LC50 do moluscicida contendo a isca para as subsequentes 72 horas causou significante recuperação dos níveis de aminoácidos livres, proteína, DNA e RNA no ovoteste da L. acuminata.

Preparation and evaluation of 5, 9-dimethyl-2-cyclopropyl-2-decanol as a penetration enhancer for drugs through rat skin

In the present study a new alcohol derivative of tetrahydrogeraniol [THG], an acyclic monoterpene, has been prepared by using Grignard reagent and methyl cyclopropyl ketone. Penetration enhancing effects of THG and the synthesized derivative 5,9-dimethyl-2-cyclopropyl-2-decanol [DICNOL] on the transdermal penetration of 5-fluorouracil [5-FU] and tramadol hydrochloride [tramadol HC1] across the excised rat skin were studied by an in vitro permeation technique using Franz diffusion cells. Azone was used as standard enhancer for comparison. DICNOL and THG significantly enhanced 5-FU and tramadol HC1 penetration through rat skin compared with the control. DICNOL enhanced the permeability of 5-FU and tramadol HC1 across full thickness skin by about 11 and 20 fold, respectively. Increased partition coefficient and diffusion coefficient values were obtained by these enhancers. The results suggest that the amount of DICNOL in the skin, especially in the stratum corneum, may be related to its penetration enhancing effects