The ethnopharmacological study of plant drugs used traditionally in Djibouti for malaria treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Djibouti was a country where malaria has been endemic for centuries. The local population use the plants as repellents or first aid for uncomplicated malaria. AIM OF THE STUDY: The aim was, for the first time, to collect and identify plants used by the local population to treat malaria and select the most interesting plants (those that are more commontly used, more available, and have fewer studies). These plants were evaluated for their antiplasmodial activity as well as their cytotoxicity on human cell lines for the most active ones. MATERIALS AND METHODS: A semi-structured questionnaire was developed for this study to collect information about the use and identity of botanical drugs used to treat malaria. The use-reports (percentage) of each plant were recorded to determine their use importance. Also, the availability status of the plants was assessed; and those in critical condition were discarded excluded from further study. Fifteen plants, out of the 41 listed, were extracted with hydro alcohol, ethyl acetate, and dichloromethane for biological testing. Chloroquine-resistant strain FcB-1 of P. falciparum and a human diploid embryonic lung cell line were used for the antiplasmodial test, and to assess the cytotoxicity for human cells respectively. Preliminary analysis of extract constituents was carried out using thin layer chromatography (TLC). RESULTS: This study identifies 41 plant taxa belonging to 32 families and records their use against malaria. Balanites rodunfolia, belonging to the Zygophyllaceae family, was the most commonly used plant, representing 44 % of use-reports. It was followed by Cadaba rodunfolia (15 %) from the Capparaceae family, and then the three species of Aloe: Aloe djiboutiensis (8.2 %), Aloe ericahenriettae (3.4 %), and Aloe rigens (3.4 %) from the Asphodelaceae family. The leaves are the most commonly used part of the plants to treat malaria, accounting for 76 % of usage. The preparation methods were decoction (52 %), maceration (29 %), and boiling (19 %). The administration routes were by oral (80 %), inhalation 19 %), and bathing (1 %). The best antiplasmodial activities were observed in the dichloromethane extracts of Cymbopogon commutatus and the ethyl acetate extracts of Aloe rigens and Terminalia brownii, with IC50 values of 9.8, 5, and 7.5 µg/mL, respectively. Their toxicity/activity levels were very favorable with selectivity indices of 5.6, 8.1, and 11.8 for C. commutatus, A. rigens, and T. Brownii, respectively. CONCLUSION: Forty-one species of botanical drugs were listed as being used to treat malaria in Djibouti. All fifteen selected species showed antiplasmodial activity (IC50 < 50 µg/mL). This work will help guide the valorization of botanical drugs used to treat malaria in Djibouti.

Antiprotozoal compounds from Mikania periplocifolia (Asteraceae).

Chagas disease, African trypanosomiasis and Leishmaniasis are neglected parasitic diseases which affect millions of people worldwide. In a previous work, we report the antiprotozoal activity of the dichloromethane extract of Mikania periplocifolia Hook. & Arn. (Asteraceae). The aim of this work was to isolate and identify the bioactive compounds present in the extract. The fractionation of the dichloromethane extract has led to the isolation of the sesquiterpene lactone miscandenin and the flavonoid onopordin, together with the sesquiterpene lactones mikanolide, dihydromikanolide and deoxymikanolide, which have previously shown antiprotozoal activity. Miscandenin and onopordin were assayed in vitro against Trypanosoma cruzi, T. brucei and Leishmania braziliensis. Miscandenin was active against T. cruzi trypomastigotes and amastigotes with IC50 values of 9.1 and 7.7 µg/ml, respectively. This sesquiterpene lactone and the flavonoid onopordin showed activity against T. brucei trypomastigotes (IC50 = 0.16 and 0.37 µg/ml) and L. braziliensis promastigotes (IC50 = 0.6 and 1.2 µg/ml), respectively. The CC50 values on mammalian cells were 37.9 and 53.4 µg/ml for miscandenin and onopordin, respectively. Besides, the pharmacokinetic and physicochemical properties of miscandenin were assessed in silico, showing a good drug-likeness profile. Our results highlight this compound as a promising candidate for further preclinical studies in the search of new drugs for the treatment of trypanosomiasis and leishmaniasis.

In vitro activity of Schinus terebinthifolius extract and fractions against Sporothrix brasiliensis.

BACKGROUND: Sporothrix brasiliensis is the causative agent of zoonotic cases of sporotrichosis in Brazil and is associated with atypical and severe presentations in cats, dogs, and humans. Sporotrichosis treatment is usually time- and cost-consuming, sometimes with poor response and host toxicity. Schinus terebinthifolius has proven efficacy against bacteria and fungi of clinical interest. OBJECTIVE: To determine the in vitro activity of S. terebinthifolius against S. brasiliensis. METHODS: Five S. brasiliensis isolates and three reference strains were subjected to a hydroethanol extract derived from the leaves of S. terebinthifolius and its fractions. The minimal inhibitory concentration (MIC) was determined using the broth microdilution method according to the M38-A2 CLSI guidelines. Also, the fungicidal/fungistatic activity of the extract and fractions was studied. FINDINGS: The crude extract of S. terebinthifolius inhibited the growth of S. brasiliensis (MIC: 0.5-1.0 µg/mL), while the partitioned extracts dichloromethane, ethyl acetate, and butanol demonstrated growth inhibition at 8 µg/mL due to a fungistatic activity. MAIN CONCLUSIONS: Due to its in vitro efficacy against S. brasiliensis and its known pharmacological safety, S. terebinthifolius is a candidate to be tested using in vivo models of sporotrichosis.

Constituents from ripe figs of Ficus vallis-choudae Delile (Moraceae) with antiplasmodial activity.

Ripe figs, barks, and wood of Ficus vallis-choudae are used in traditional medicine against several conditions including nausea and malaria. However, its use is still to be scientifically documented and validated. Hence, the aim of the present work was to evaluate the antiplasmodial activity of the dichloromethane-methanol (DCM-MeOH (1:1)) crude extract, their hexane, dichloromethane, ethyl acetate, and methanoli fractions, as well as the isolated chemical constituents. The chemical study of the DCM-MeOH (1:1) crude extract of F. vallis-choudae figs led to the isolation of fifteen (15) known compounds identified based on their spectroscopic data [one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR), mass spectrometry] and by comparison of these data with those reported in the literature. Some of the isolated compounds were assessed in vitro for their antiplasmodial activity against Plasmodium falciparum chloroquine-sensitive 3D7 (Pf3D7) and multidrug-resistant Dd2 strains. The dichloromethane fraction exhibited very good antiplasmodial activity against both strains with IC50 values of 13.86 µg/mL and 8.18 µg/mL, respectively. Among the tested compounds, wighteone (2) was the most active against P. falciparum 3D7 (IC50 = 24.6 ± 1.5 µM) and Dd2 (IC50 = 11.9 ± 2.4 µM) strains. The obtained results could justify the traditional uses of F. vallis-choudae against malaria. Wighteone appears to be the most active ingredient. However, further consideration of this compound as starting point for antimalarial drug discovery will depend upon its selectivity of action towards Plasmodium parasites. HIGHLIGHTS: • 15 (fifteen) compounds were isolated from the dichloromethane-methanol extract of Ficus vallis-choudae. • Their structures were determined on the basis of their spectroscopic data. • The dichloromethane fraction showed promising activities on the Pf3D7 and PfDd2 strains with IC50 values of 13.86 and 8.18 µg/mL, respectively. • Wighteone was the most active compound against PfDd2 (IC50 = 11.9 ± 2.4 µM).

In vitro and in vivo anti-cancer activity of dichloromethane fraction of Triticum aestivum sprouts.

Triticum aestivum sprouts (TA) contain significant amounts of chlorophyll, minerals, enzymes, and other functional entities. Furthermore, TA extracts have been shown to possess anti-obesity, anti-diabetic and hepatoprotective effects and are believed to help blood flow, digestion, and general detoxification of the body. In this study, the mechanism underlying the anti-cancer effects of a dichloromethane fraction of TA (TDF) was investigated in vitro and in vivo. In vitro study was done by examining cancer cells growth, morphological changes, cell cycles, expressions of death receptors and apoptosis-linked proteins in wide range of human cancer cell lines. To investigate the effect of TDF in vivo, C57BL/6 mice were injected with B16 melanoma cells and orally administered TDF. TDF markedly inhibited cancer cell growth and induced cellular morphological alterations, cell cycle arrest and apoptosis, and enhanced the expressions of death receptors (DR)-4, 5, and 6 in cell lines. In addition, TDF regulated the expressions mitochondrial apoptosis-linked proteins and induced caspase-dependent cell death. It also significantly enhanced phosphorylation of ERK1/2 and JNK, but not p38, whereas inhibited the activation of NF-κB in cancer cells. In our mouse model, TDF significantly suppressed B16 melanoma growth, to an extent similar to cisplatin (reference control) and augmented immunomodulatory cytokines. In brief, this study presents the mechanism responsible for the anti-cancer effects of TDF in vitro and in vivo.

Antinociceptive and anti-inflammatory activities of the Jatropha isabellei dichloromethane fraction and isolation and quantitative determination of jatrophone by UFLC-DAD.

CONTEXT: Jatropha isabellei Müll. Arg. (Euphorbiaceae) has been used in the traditional medicine to treat arthritis. OBJECTIVE: To evaluate the anti-inflammatory and antinociceptive activities of the dichloromethane fraction (DFJi) from underground parts of J. isabellei, and to develop an analytical method to quantify the diterpene jatrophone. MATERIALS AND METHODS: Anti-inflammatory and antinociceptive activities of the DFji were determined by an acute arthritis model through assessment of the paw elevation time (PET) and articular diameter (AD) of Wistar rats treated orally (50, 100 or 200 mg/kg in a single-dose), and intravenously (0.1, 1, 10, 25 or 50 mg/kg in a bolus administration). The isolation of jatrophone from the DFji was carried out and confirmed by spectroscopic techniques. A UFLC-DAD method was developed and validated. RESULTS: When orally administered, the highest dose (200 mg/kg) of DFJi was able to significantly reduce the PET to 24.8 ± 1.4 s (p < 0.01), when compared with the control group (33.7 ± 1.8 s). The administration of the intravenous dose of 10 mg/kg reduced the PET to 14.8 ± 0.3 s (p < 0.001). The oral and intravenous administration of the DFJi at dose of 200 and 10 mg/kg significantly prevented the formation of edema, reducing the AD in 25.3% and 32.5% (p < 0.01), respectively. The UFLC-DAD method allowed the quantification of jatrophone, which was found to be around 90 µg/mg of fraction. DISCUSSION AND CONCLUSION: The DFJi displayed antinociceptive and antiedematogenic activities, representing a promising plant product for the arthritis treatment.

Estudio analítico de MTA-Angelus(R) y Biodentine(R) con técnicas SPME-GCMS y AFM / Analytical studies of MTA Angelus and Biodentine materials by SPME-GCMS and AFM techniques

Introducción: El objetivo del estudio fue obtener información analítica sobre la pureza química y homogeneidad de los materiales de uso endodóntico MTA-Angelus® y Biodentine®. Material y métodos: Se analizaron ambos materiales mediante la técnica de microextracción en fase sólida (SPME), combinada con cromatografía de gases acoplada a espectrómetro de masas (GC-MS) para separar e identificar los compuestos orgánicos volátiles (COVs) contenidos. Por otro lado, utilizando microscopía de fuerza atómica (AFM) se estudió la topografía y rugosidad de las superficies de dientes tras la realización de obturaciones a retro in vitro con ambos cementos. Resultados: Se identificaron diversos COVs en ambos materiales mediante SPME GCMS. Entre los compuestos detectados, destacaron el alfa - metilestireno (presente en MTA-Angelus(R)) y el diclorometano (detectado tanto en MTA-Angelus(R) como en Biodentine(R)) que son potencialmente tóxicos y perjudiciales para la salud. Los resultados del estudio con AFM demostraron la existencia de diferencias en la superficie de los cementos. Las imágenes y rugosidad finales fueron más homogéneas en Biodentine®. Conclusiones: Los cementos endodónticos MTA-Angelus® y Biodentine® contienen ciertos compuestos perjudiciales para la salud. Se evidencia la necesidad de un control más exigente de los riesgos para la salud de estos. El estudio AFM mostró que Biodentine® presenta un superficie más homogénea (AU)

Introduction: The aim of this research was to obtain analytical information about the chemical purity and the homogeneity of the endodontic materials MTA Angelus(TM) and Biodentine(TM). Methods: On the one hand, solid phased microextraction-gas chromatography-mass spectrometry (SPME-GCMS) was carried out with MTAAngelus and Biodentine materials, isolating and identifying the contained volatile organic compounds (VOCs). On the other hand, atomic force microscopy (AFM) was used to study topography and roughness aspects on teeth after in vitro root end fillings with both materials. Results: First, several VOCs have been detected by SPME-GCMS in both materials. It is noteworthy alpha methylstirene (identified in MTA Angelus(TM)) or dichloromethane (detected both in MTA Angelus(TM) and Biodentine(TM)) due to its potential toxicity and harmful health effects. Secondly, AFM study showed differences among the surface materials. Final images and roughness results showed more homogeneous cement in the Biodentine product. Conclusions: Endodontic cements MTAAngelus and Biodentine contains some harmful health compounds. The need of a stricter health risks control of both materials became evident. AFM study demonstrated a more homogenous surface in Biodentine (AU)

Xanthone-rich dichloromethane fraction of Securidaca inappendiculata, the possible antirheumatic material base with anti-inflammatory, analgesic, and immunodepressive effects.

CONTEXT: Securidaca inappendiculata Hassk. is an traditional Chinese medicine curing rheumatoid arthritis, but there is a lack of reports on material base research. OBJECTIVE: To find the active fraction of S. inappendiculata contributing the most to antirheumatic activity. MATERIALS AND METHODS: Prior to assays in vivo, mice were treated with different fractions from S. inappendiculata for 5 d at doses relative to 10, 5, and 2.5 g/kg of crude drug. Hot plate test and carrageenan-induced paw edema test were used to investigate analgesic and anti-inflammatory activities. PGE2 levels in inflammatory paws were determined by a colorimetric method. Carbon clearance test in vivo and lymphocyte transformation test in vitro were employed to assess the immune regulation activity. HPLC was used to explore the main compounds in the active fraction. RESULTS: All the fractions, especially the dichloromethane fraction (SID), alleviated inflammation. High dose of SID (112 mg/kg) inhibited paw swelling by 63.1%, and decreased PGE2 level to 38 ng/mL. The ethyl acetate fraction (SIE) and SID suppressed the carbon clearance rate (K = 0.044, 0.038 for high dose) efficiently. All fractions hindered the transformation and proliferation of lymphocyte, and prolonged the reaction time of rats in the hot plate test. The concentrations of two typical xanthones: 2-hydroxyl-1,7-dimethoxyl-xanthone and 1,7-dihydroxyl-xanthone in SID were 0.93% and 1.19%, respectively, by HPLC analysis. CONCLUSION: SID exhibited significant anti-inflammatory, analgesic, and immunodepressive effects in vivo and vitro, and deemed as the main material base for the antirheumatic activity.

In vitro and in vivo antileishmania activity of sesquiterpene lactone-rich dichloromethane fraction obtained from Tanacetum parthenium (L.) Schultz-Bip.

The discovery of new treatments for neglected diseases, including leishmaniasis, is a substantial challenge for scientific research. Plant extracts have shown potential in the selective treatment of tropical diseases. The present study evaluated the in vitro and in vivo antileishmania effects of a sesquiterpene lactone-rich dichloromethane fraction (DF) obtained from the aerial parts of Tanacetum parthenium (L.) Schultz-Bip. In vitro studies of the DF indicated an IC50 of 2.40±0.76 µg mL(-1) against the promastigote form and 1.76±0.25 µg mL(-1) against the axenic amastigote form of Leishmania amazonensis. In vivo intramuscular treatment with DF decreased the growth and size of footpad lesions in mice. The DF also significantly decreased the parasite population compared with animals that were treated with the reference drug. Plasma malondialdehyde levels were increased slightly by the DF, attributable to its parthenolide-rich composition that causes cellular apoptosis, compared with the control group, demonstrating treatment efficacy without toxicity or genotoxicity. Because the isolation and purification of plant compounds are costly and time-consuming and generate low yields, extract fractions, such as the DF studied herein, represent a promising alternative for the treatment of leishmaniasis.

Therapeutic effects of dichloromethane fraction of Securidaca inappendiculata on adjuvant-induced arthritis in rat.

ETHNOPHARMACOLOGICAL RELEVANCE: Securidaca inappendiculata (SI) is a traditional antirheumatic medicine used in China. The present study was designed to investigate the therapeutic efficacy of dichloromethane fraction of SI (SID) at three different doses on adjuvant induced arthritis (AA) rats. METHODS: Arthritis severity was evaluated by arthritic score, body weight loss, paw circumference, histological changes and hyperplasia of lymphatic tissues. Serum samples were collected for estimation of superoxide dismutase (SOD), glutathione (GSH), hydroxy radical (OH·), nitric oxide (NO), malondialdehyde (MDA), N-acetyl glucosaminidase (NAG), sialic acid (SA), alkaline phosphatase (ALP), aspartate transaminase (AST) and alanine transaminase (ALT). The levels of GSH, MDA, NAG and SA in liver were also assessed. The levels of interleukin-1 (IL-1), tumor necrosis factor alpha (TNF-α), monocyte chemotactic protein 1 (MCP-1) and vascular endothelial growth factor (VEGF) were determined using ELISA method. Another portion of blood was used for total and differential leucocyte counts. RESULTS: Administration with SID (at high dose with 100 mg/kg) significantly ameliorated the AA severity, suggested by the modulatory effects on body weight loss, paw swelling, hyperplasia of lymphatic tissues and synovial membrane, neutrocytosis and lymphocytosis. It also decreased levels of NO, MDA and OH·, restored SOD and GSH levels in serum. The abnormal increased levels of AST, ALT, ALP, NAG and SA significantly were reverted (compared with AA rats, P<0.01). A similar result was observed in livers. Levels of IL-1, TNF-α, MCP-1 and VEGF were reduced dramatically by SID too. CONCLUSION: The results suggest SID possesses substantial anti-arthritic activity. The therapeutic efficacy may be due to immumodepressive effects, cytokines regulation, increasing membrane stability and antioxidantive activity.

Hypotensive mechanism of the extracts and artemetin isolated from Achillea millefolium L. (Asteraceae) in rats.

Traditional uses of Achillea millefolium L. (Asteraceae) include the treatment of cardiovascular diseases. In the present study, we used anesthetized rats to assess the hypotensive effect of a hydroethanolic extract (HEAM), and its dichloromethane (DCM), ethyl acetate (EA), butanolic (BT), and dichloromethane-2 (DCM-2) fractions, besides the flavonoid artemetin, isolated from A. millefolium. The oral administration of HEAM (100-300 mg/kg), DCM (20mg/kg), DCM-2 (10-30 mg/kg), but not EA (10 mg/kg) and BT (50 mg/kg) fractions significantly reduced the mean arterial pressure (MAP) of normotensive rats. The phytochemical analysis by NMR (1)H of DCM and DCM-2 fractions revealed high amounts of artemetin, that was isolated and administered by either oral (1.5 mg/kg) or intravenous (0.15-1.5 mg/kg) routes in rats. This flavonoid was able to dose-dependently reduce the MAP, up to 11.47 ± 1.5 mmHg (1.5 mg/kg, i.v.). To investigate if artemetin-induced hypotension was related to angiotensin-converting enzyme inhibition, we evaluated the influence of this flavonoid on the vascular effects of both angiotensin I and bradykinin. Intravenous injection of artemetin (0.75 mg/kg) significantly reduced the hypertensive response to angiotensin I while increased the average length of bradykinin-induced hypotension. Artemetin (1.5 mg/kg, p.o.) was also able to reduce plasma (about 37%) and vascular (up to 63%) ACE activity in vitro, compared to control group. On the other hand, artemetin did not change angiotensin II-induced hypertension. Our study is the first showing the hypotensive effects induced by the extract and fractions obtained from A. millefollium. In addition, our results disclosed that this effect may be, at least in part, associated with high levels of artemetin and its ability to decrease angiotensin II generation in vivo, by ACE inhibition.

Efecto sedante del triterpeno cicloartanol obtenido de las hojas de Pedilanthus tithymaloides (L) Poit / Sedative effect of cycloartanol triterpene obtained from Pedilanthus tithymaloides (L) Poit leaves

INTRODUCCIÓN: Pedilanthus tithymaloides (L.) Poit (ítamo real) es una planta ampliamente distribuida en Cuba y constituye una fuente sustentable para la obtención de principios activos. OBJETIVO: evaluar el efecto neurosedante del triterpeno cicloartanol, obtenido a partir del extracto diclorometano de las hojas de esta planta. MÉTODOS: se emplearon los modelos de conducta exploratoria, sueño barbitúrico, convulsiones inducidas por pentilenotetrazol y estereotipias inducidas por anfetamina. El cicloartanol fue administrado por vía oral 30 min antes de cada ensayo. RESULTADOS: se obtuvo inhibición de la conducta exploratoria a dosis superiores a 100 mg/kg, inhibición de las estereotipias inducidas por anfetamina a dosis de 200 mg/kg, se potenció el sueño inducido por tiopental a dosis superiores a 50 mg/kg y no se observó protección frente a las convulsiones inducidas por pentilenotetrazol. CONCLUSIONES: los resultados obtenidos en los diferentes ensayos muestran un efecto depresor del sistema nervioso central en animales tratados con cicloartanol, sin embargo, el efecto sedante no parece estar mediado por el sistema GABAérgico, tampoco se observaron efectos antidepresivos

INTRODUCTION: Pedilanthus tithymaloides (L.) Poit (known as ítamo real) is a widely distributed plant in Cuba and at the same time a sustainable source of active principles. OBJECTIVE: to evaluate the neurosedative effect of cycloartanol triterpene from dichlorometane extract obtained from Pedilanthus tithymaloides (L.) Poit leaves. METHODS: models based on exploratory behaviour, barbiturate-induced sleep, pentilenotetrazol-induced seizures and amphetamine-induced stereotyped behaviours were used. Cyclocartanol was orally administered 30 minutes before every test. RESULTS: exploratory behaviour was inhibited at over 50 mg/kg dose; amphetamine-induced stereotyped behaviours were inhibited at 200 mg/kg dose; thiopental-induced sleep was potentiated at over 50 mg/kg dose, however, protection against pentilenotetrazole-induced seizures was not observed. CONCLUSIONS: the results of several tests indicate a depressing effect on the central nervous system of laboratory animal treated with Cycloartanal; however, the sedative effect does not seem to be mediated by the GABAergic system. Anti-depressive effects were not observed either

Efecto sedante del triterpeno cicloartanol obtenido de las hojas de Pedilanthus tithymaloides (L) Poit / Sedative effect of cycloartanol triterpene obtained from Pedilanthus tithymaloides (L) Poit leaves

INTRODUCCIÓN: Pedilanthus tithymaloides (L.) Poit (ítamo real) es una planta ampliamente distribuida en Cuba y constituye una fuente sustentable para la obtención de principios activos. OBJETIVO: evaluar el efecto neurosedante del triterpeno cicloartanol, obtenido a partir del extracto diclorometano de las hojas de esta planta. MÉTODOS: se emplearon los modelos de conducta exploratoria, sueño barbitúrico, convulsiones inducidas por pentilenotetrazol y estereotipias inducidas por anfetamina. El cicloartanol fue administrado por vía oral 30 min antes de cada ensayo. RESULTADOS: se obtuvo inhibición de la conducta exploratoria a dosis superiores a 100 mg/kg, inhibición de las estereotipias inducidas por anfetamina a dosis de 200 mg/kg, se potenció el sueño inducido por tiopental a dosis superiores a 50 mg/kg y no se observó protección frente a las convulsiones inducidas por pentilenotetrazol. CONCLUSIONES: los resultados obtenidos en los diferentes ensayos muestran un efecto depresor del sistema nervioso central en animales tratados con cicloartanol, sin embargo, el efecto sedante no parece estar mediado por el sistema GABAérgico, tampoco se observaron efectos antidepresivos(AU)

INTRODUCTION: Pedilanthus tithymaloides (L.) Poit (known as ítamo real) is a widely distributed plant in Cuba and at the same time a sustainable source of active principles. OBJECTIVE: to evaluate the neurosedative effect of cycloartanol triterpene from dichlorometane extract obtained from Pedilanthus tithymaloides (L.) Poit leaves. METHODS: models based on exploratory behaviour, barbiturate-induced sleep, pentilenotetrazol-induced seizures and amphetamine-induced stereotyped behaviours were used. Cyclocartanol was orally administered 30 minutes before every test. RESULTS: exploratory behaviour was inhibited at over 50 mg/kg dose; amphetamine-induced stereotyped behaviours were inhibited at 200 mg/kg dose; thiopental-induced sleep was potentiated at over 50 mg/kg dose, however, protection against pentilenotetrazole-induced seizures was not observed. CONCLUSIONS: the results of several tests indicate a depressing effect on the central nervous system of laboratory animal treated with Cycloartanal; however, the sedative effect does not seem to be mediated by the GABAergic system. Anti-depressive effects were not observed either(AU)

Anti-inflammatory activity of methylene chloride fraction from Glehnia littoralis extract via suppression of NF-kappa B and mitogen-activated protein kinase activity.

Glehnia littoralis (Umbelliferae) has been used traditionally in Korean, Japanese, and Chinese medicine for the treatment of immune-related diseases; however, its anti-inflammatory activity and underlying mechanism remain to be defined. We investigated the anti-inflammatory effect and inhibitory mechanism on inflammation by the methylene chloride fraction from Glehnia littoralis extract (MCF-GLE), which was more effective than Glehnia littoralis extract (GLE). MCF-GLE inhibited 12-O-Tetradecanoyl-phorbol-13-acetate (TPA)-induced inflammation in an inflammatory edema mouse model. Also, MCF-GLE strongly inhibited the releases of nitric oxide (NO), prostaglandin E(2) (PGE(2)), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta) and significantly suppressed the mRNA and protein expression of inducible nitric oxide synthase and cyclooxygenase-2 in lipopolysaccharide-stimulated RAW 264.7 macrophage cells in a dose-dependent manner. Furthermore, MCF-GLE suppressed NF-kappaB activation and IkappaB-alpha degradation. MCF-GLE also attenuated the activation of ERK and JNK in a dose-dependent manner. These results indicate that MCF-GLE has an inhibitory effect on the in vivo and in vitro inflammatory reaction and is a possible therapeutic agent. Our results suggest that the anti-inflammatory properties of MCF-GLE may result from the inhibition of pro-inflammatory mediators, such as NO, PGE(2), TNF-alpha, and IL-1beta via suppression of NF-kappaB- and mitogen-activated protein kinases-dependent pathways.

Cytotoxicity and antitumoral activity of dichloromethane extract and its fractions from Pothomorphe umbellata.

The cytotoxicity of the dichloromethane crude extract (DCE), obtained from the aerial parts of Pothomorphe umbellata (L.) Miq (Piperaceae), was evaluated against nine human cancer cell lines (MCF-7, NCI-ADR/RES, OVCAR-3, PC-3, HT-29, NCI-H460, 786-O, UACC-62, K-562). The DCE presented antiproliferative activity with good potency against all cell lines at low concentrations (between 4.0 and 9.5 microg/mL) and with selectivity (1.55 microg/mL) for the leukemia cell line (K-652). DCE (100, 200, 300 and 400 mg/kg, ip) was also evaluated in the Ehrlich ascites tumor model. Both the survival number and the life span of the animals that died increased by at least 45 and 50%, respectively (8 animals per group), demonstrating P. umbellata extract potential anticancer activity. The results of the in vivo antitumor activity prompted the fractionation of the crude extract. The crude extract was submitted to dry column chromatography with dichloromethane-methanol (99:1). The column effluent fractions were extracted with methanol, dried under vacuum yielding fractions FR1 (less polar), FR2 (medium polarity), and FR3 (polar), which were analyzed for their growth inhibition or cytotoxic properties by a 48-h sulforhodamine B cell viability assay by measuring the total protein content. FR1 demonstrated high potency and cytotoxicity, a result compatible with the high toxicity of oxalic acid; FR2, containing 4-nerolidylcathecol, presented the lowest cytotoxic activity compared to the other two fractions but with selectivity for prostate cancer cell line; FR3, containing a mixture of steroids described in the literature as possessing various biological activities, also presented potent anticancer in vitro activity. These results suggest that P. umbellata DCE in vivo antitumor activity may be a consequence of the activity of different active principles.

Cytotoxicity and antitumoral activity of dichloromethane extract and its fractions from Pothomorphe umbellata

The cytotoxicity of the dichloromethane crude extract (DCE), obtained from the aerial parts of Pothomorphe umbellata (L.) Miq (Piperaceae), was evaluated against nine human cancer cell lines (MCF-7, NCI-ADR/RES, OVCAR-3, PC-3, HT-29, NCI-H460, 786-O, UACC-62, K-562). The DCE presented antiproliferative activity with good potency against all cell lines at low concentrations (between 4.0 and 9.5 µg/mL) and with selectivity (1.55 µg/mL) for the leukemia cell line (K-652). DCE (100, 200, 300 and 400 mg/kg, ip) was also evaluated in the Ehrlich ascites tumor model. Both the survival number and the life span of the animals that died increased by at least 45 and 50 percent, respectively (8 animals per group), demonstrating P. umbellata extract potential anticancer activity. The results of the in vivo antitumor activity prompted the fractionation of the crude extract. The crude extract was submitted to dry column chromatography with dichloromethane-methanol (99:1). The column effluent fractions were extracted with methanol, dried under vacuum yielding fractions FR1 (less polar), FR2 (medium polarity), and FR3 (polar), which were analyzed for their growth inhibition or cytotoxic properties by a 48-h sulforhodamine B cell viability assay by measuring the total protein content. FR1 demonstrated high potency and cytotoxicity, a result compatible with the high toxicity of oxalic acid; FR2, containing 4-nerolidylcathecol, presented the lowest cytotoxic activity compared to the other two fractions but with selectivity for prostate cancer cell line; FR3, containing a mixture of steroids described in the literature as possessing various biological activities, also presented potent anticancer in vitro activity. These results suggest that P. umbellata DCE in vivo antitumor activity may be a consequence of the activity of different active principles.

Orthopaedic metal devices coated with a novel antiseptic dye for the prevention of bacterial infections.

Gendine is a novel antiseptic dye with broad-spectrum antimicrobial activity that may be used to coat plastics and metal devices. Our objective was to determine the efficacy of gendine-coated orthopaedic metal devices in preventing methicillin-resistant Staphylococcus aureus (MRSA) colonisation. Stainless steel and titanium Schanz rods were coated with gendine. The zone of inhibition (ZoI) around the rods with and without gamma-irradiation was determined by a modified Kirby-Bauer method. A previously published bioprosthetic biofilm colonisation model, modified Kuhn's method, was used to determine the adherence of MRSA to coated and uncoated rods, with and without irradiation, after insertion into bovine bone and after 3 months shelf life followed by 2 weeks of immersion in serum. The gendine-coated Schanz metal rods showed a net ZoI of 16 mm against MRSA before and after irradiation. Gendine-coated rods showed no biofilm formation (0 colony-forming units (CFU)), which was a significant reduction (P<0.001) compared with uncoated controls (>5000 CFU). Coated rods exposed to high-dose gamma-irradiation and coated rods drilled into bone also showed significant efficacy (P<0.001) in preventing biofilm adherence. After 2 weeks, gendine-coated rods maintained significant durability (P<0.01), resulting in 90% reduction in MRSA biofilm adherence compared with uncoated control rods. Results indicate that gendine-coated metal rods are highly efficacious in the prevention of MRSA biofilm.

Cytotoxic activity of the dichloromethane fraction from Vernonia scorpioides (Lam.) Pers. (Asteraceae) against Ehrlich's tumor cells in mice.

Vernonia scorpioides has been widely used in Brazil to treat skin problems and chronic wounds, such as ulcers of the lower limbs and diabetic lesions. In the present study, we investigated the effect of a dichloromethane (DCM) fraction of V. scorpioides leaf extract on Ehrlich ascitic and solid tumor-bearing mice. The animals were treated once a day with the DCM fraction at a concentration of 5 mg/kg, administered ip during and after the development of the tumor. The lifespan, weight, number and type of leukocytes, number of tumor cells, volume of solid and ascitic tumors were measured. The development of the tumor with pre-treated tumor cells in vitro with the DCM fraction (5 mg/kg) was analyzed and the animals were sacrificed after 7 days. The DCM fraction (5 mg/kg) totally inhibited tumor development when in direct contact with tumor cells, and also ascitic tumor development with in vitro treatment or when administered ip, in loco (after 7 days). Animals treated with the DCM fraction increased their lifespan ca. 2 weeks and maintained their body weight for 30 days. When applied immediately after the inoculation of the tumor cells in vivo, it totally abolished tumor development, with tumor development only decreasing when treatment was started 3 days after the tumor challenge. These data suggest an antineoplastic activity of the fraction. Oral or ip administration of DCM fraction (5 mg/kg) for 7 days did not reduce the solid tumor volume. The cytotoxic activity described here differs from the conventional immune suppressing profile of standard chemotherapy because it increases neutrophil influx to the peritoneal cavity. These results show that, besides exhibiting a tumoricidal activity, the DCM fraction also exhibits inflammatory activity.

Antidiabetic and hypolipidaemic effects of a methanol/methylene-chloride extract of Laportea ovalifolia (Urticaceae), measured in rats with alloxan-induced diabetes.

A decoction of the leaves of Laportea ovalifolia is widely used in Cameroon for the treatment of several illnesses, including diabetes mellitus. The antidiabetic and hypolipidaemic effects of a methanol/methylene-chloride extract of the aerial parts of L. ovalifolia have now been investigated, in normal rats and rats with diabetes induced by the intraperitoneal injection of alloxan (at 150 mg/kg bodyweight). In the diabetic rats, 2 weeks of daily, intragastric treatment with the L. ovalifolia extract not only produced a significant reduction in the fasting serum glucose concentrations but also lowered the serum concentrations of total cholesterol, triglycerides, and low-density-lipoprotein cholesterol, lowered the ratio of total cholesterol to high-density-lipoprotein (HDL) cholesterol, and increased the serum concentration of HDL cholesterol. At least in rats with alloxan-induced diabetes, the methanol/methylene-chloride extract of L. ovalifolia therefore appears to possess antidiabetic and hypolipidaemic properties.

Evaluation of the antiedematogenic activity of artemetin isolated from Cordia curassavica DC.

The species Cordia curassavica (Boraginaceae), known popularly as "erva baleeira", is used in folk medicine for the treatment of several inflammatory processes and as a healing agent. The objective of the present study was to evaluate the antiedematogenic activity of crude dichloromethane extracts of Cordia curassavica and of the artemetin-enriched fraction. The crude extract and artemetin fraction were tested in the model of carrageenin-induced paw edema in male Swiss mice (25-30 g). The crude dichloromethane extract (300 and 1000 mg/kg, po, N = 6) showed significant antiedematogenic activity, reducing the edema by 42, 57 and 45% and 46, 62 and 69%, 3, 4 and 5 h after carrageenin administration, respectively. Indomethacin (10 mg/kg, po, N = 6) reduced the edema by 45 and 48%, after 4 and 5 h, but the artemetin-enriched fraction (30, 100 and 300 mg/kg, po, N = 6) had no activity. The dichloromethane extract (300 and 1000 mg/kg, po, N = 6) also showed antinociceptive activity by reducing acetic acid-induced writhing in mice from 37.1 +/- 2.28 (control) to 17.3 +/- 1.34 and 13.2 +/- 1.44, respectively, but had no activity in the hot-plate test.