Evidence for the Involvement of Cytokines Modulation and Prokinetic Properties in Gastric Ulcer Healing Effects of Helicteres sacarolha A. St.-Hil. A. Juss.

Preparations of Helicteres sacarolha (Malvaceae) leaves and roots are used in the form of decoction, infusion or maceration, to treat gastrointestinal disturbances, among others. Studies supporting some of its ethnomedicinal uses are still incipient. The present study aimed to investigate it potential effect on chronic ulcer, ulcerative colitis and possible prokinetic activities as part of its mechanism of action. The powdered leaves of Helicteres sacarolha (HEHs) was prepared by maceration in 70 % hydroethanolic solution. Its qualitative phytochemical constituents were investigated by direct flow injection analysis coupled to atmospheric pressure chemical ionization ion trap tandem mass spectrometry (FIA-APCI-IT-MSn ). The gastric ulcer healing effect was evaluated in acetic acid induced chronic ulcer in mice and the lesions were evaluated, including analysis of blood plasma cytokine levels. The prokinetic properties (gastric emptying and intestinal transit) were carried out in mice. Potential anti-ulcerative colitis activity was evaluated in rats using 2,4,6-trinitrobenzenesulfonic acid (5 % TNBS) -induced colitis. All animal experiments were carried out at the doses of 20, 50 and 250 mg/kg (p.o.). Eight compounds were putatively identified, specifically lariciresinol, and its derivatives, kaempferol derivatives and Tricin-O-Glc. The extract promoted increased gastric ulcer healing at all doses tested. Modulation of the cytokines involved inhibition of some key pro-inflammatory cytokines with maximum effect on IL-1ß (70 %, 50 mg/kg, p<0.05), TNF-α (79 %, 20 mg/kg, p<0.01), and in the anti-inflammatory cytokines, namely IL-10 (57 %, 50 mg/kg, p<0.05) and IL-17 (79 %, only at 50 mg/kg, p<0.05). Histological findings demonstrated a mitigated inflammatory activity, and tissues undergoing regeneration. HEHs treatment caused delayed gastric emptying, and increased intestinal transit, but had no effect in the experimentally induced ulcerative colitis. We report for the first time putatively the presence of Lariciresinol and tricin derivatives from the hydroethanolic leaves extract of H. sacarolha. Its possible mechanism of actions of gastric ulcer healing involves cytokines modulation, mitigation of inflammatory response and tissue regeneration and provoked opposing effect in the gastrointestinal system. The present study demonstrates the therapeutic potential of H. sacarolha leaves used in Brazilian ethnomedicine in the treatment of chronic gastric ulcer.

Avaliação da citotoxicidade in vitro e genotoxicidade ex-vivo em compostos da Pavonia glazioviana Gürke (Malvaceae / Evaluation of in vitro cytotoxicity and ex-vivo genotoxicity in compounds from Pavonia glazioviana Gürke(Malvaceae)

Introdução: as terapias alternativas que utilizam plantas medicinais e fitoterápicos são bastante comuns no Brasil. Dentre várias espécies vegetais brasileiras utilizadas em terapias destacam-se as espécies da família Malvaceae. Objetivos: o presente estudo teve como objetivo avaliar a citotoxicidade in vitro e a genotoxicidade ex-vivo em compostos da Pavonia glazioviana Gürke espécie brasileira pertencente à família Malvaceae. Metodologia: métodos in vitro foram utilizados para verificar o potencial citotóxico por meio de ensaios hemolíticos e anti-hemolíticos e da análise genotóxica ex-vivo. O Extrato Etanólico Bruto (EEB) e Fração Clorofórmico (FC) foram obtidos na amostra vegetal utilizada neste estudo. Resultados: os produtos EEB-Pg e FC-Pg apresentaram baixo efeito citotóxico apenas nas concentrações de 50 e 100 µg / mL. As amostras expostas às concentrações de 500 e 1000 µg / mL apresentaram índice hemolítico alto a moderado com lise superior a 60%. Foi descrito efeito anti-hemolítico moderado em todas as amostras tratadas com 500 e 1000 µg / mL, com hemólise < 60%. Além disso, os compostos mostraram baixo efeito genotóxico ex-vivo, com um índice geral de células normais superior a 84% em todas as concentrações. Conclusões: os resultados sugerem um baixo perfil tóxico dos compostos obtidos da espécie Pavonia glazioviana, indicando limites seguros para o uso desses produtos naturais.

Introduction: alternative therapies using medicinal plants and herbal medicines are quite common in Brazil. Among several Brazilian plant species used in therapies, the species of the Malvaceae family stand out. Objetctives: the present study aimed to evaluate the in vitro cytotoxicity and ex-vivo genotoxicity in compounds of the Brazilian Pavonia glazioviana Gürke belonging to the Malvaceae family. Methodology: in vitro methods were used to verify the cytotoxic potential through hemolytic and antihemolytic assays and the ex-vivo genotoxic analysis. The Crude Etanolic Extract (CEE) and Cloroformic Fraction (CF) was obtained in vegetal sample used on this study. Results: the CEE-Pg and CF-Pg products only showed a low cytotoxic effect at the concentrations of 50 and 100 µg/mL. The exposure to the concentrations of 500 and 1000 µg/mL showed a high to moderate hemolytic index with lysis higher than 60%. A moderate anti-hemolytic effect was described in all samples treated with 500 and 1000 µg/mL, with hemolysis <60%. In addition, the compounds showed low ex-vivo genotoxic effect with a general index of normal cells greater than 84% at all concentrations. Conclusion: the results suggest a low toxic profile of the compounds obtained from the Pavonia glazioviana Gürke species belonging to the Malvaceae family, indicating safe limits for the use of these natural products.

Determination of hemicellulose, cellulose, holocellulose and lignin content using FTIR in Calycophyllum spruceanum (Benth.) K. Schum. and Guazuma crinita Lam.

Capirona (Calycophyllum spruceanum (Benth.) K. Schum.) and Bolaina (Guazuma crinita Lam.) are fast-growing Amazonian trees with increasing demand in timber industry. Therefore, it is necessary to determine the content of cellulose, hemicellulose, holocellulose and lignin in juvenile trees to accelerate forest breeding programs. The aim of this study was to identify chemical differences between apical and basal stem of Capirona and Bolaina to develop models for estimating the chemical composition using Fourier transform infrared (FTIR) spectra. FTIR-ATR spectra were obtained from 150 samples for each species that were 1.8 year-old. The results showed significant differences between the apical and basal stem for each species in terms of cellulose, hemicellulose, holocellulose and lignin content. This variability was useful to build partial least squares (PLS) models from the FTIR spectra and they were evaluated by root mean squared error of predictions (RMSEP) and ratio of performance to deviation (RPD). Lignin content was efficiently predicted in Capirona (RMSEP = 0.48, RPD > 2) and Bolaina (RMSEP = 0.81, RPD > 2). In Capirona, the predictive power of cellulose, hemicellulose and holocellulose models (0.68 < RMSEP < 2.06, 1.60 < RPD < 1.96) were high enough to predict wood chemical composition. In Bolaina, model for cellulose attained an excellent predictive power (RMSEP = 1.82, RPD = 6.14) while models for hemicellulose and holocellulose attained a good predictive power (RPD > 2.0). This study showed that FTIR-ATR together with PLS is a reliable method to determine the wood chemical composition in juvenile trees of Capirona and Bolaina.

Metabolic Profiling of Vasorelaxant Extract from Malvaviscus arboreus by LC/QTOF-MS.

We aimed to develop a standardized methodology to determine the metabolic profile of organic extracts from Malvaviscus arboreus Cav. (Malvaceae), a Mexican plant used in traditional medicine for the treatment of hypertension and other illnesses. Also, we determined the vasorelaxant activity of these extracts by ex vivo rat thoracic aorta assay. Organic extracts of stems and leaves were prepared by a comprehensive maceration process. The vasorelaxant activity was determined by measuring the relaxant capability of the extract to decrease a contraction induced by noradrenaline (0.1 µM). The hexane extract induced a significant vasorelaxant effect in a concentration- and endothelium-dependent manner. Secondary metabolites, such as polyunsaturated fatty acids, terpenes and one flavonoid, were annotated by liquid chromatography/quadrupole time-of-flight mass spectrometry (LC/QTOF-MS) in positive ion mode. This exploratory study allowed us to identify bioactive secondary metabolites from Malvaviscus arboreus, as well as identify potentially-new vasorelaxant molecules and scaffolds for drug discovery.

Insights on the Larvicidal Mechanism of Action of Fractions and Compounds from Aerial Parts of Helicteres velutina K. Schum against Aedes aegypti L.

Viral diseases transmitted by the female Aedes aegypti L. are considered a major public health problem. The aerial parts of Helicteres velutina K. Schum (Sterculiaceae) have demonstrated potential insecticidal and larvicidal activity against this vector. The objective of this research was to investigate the mechanisms of action involved in the larvicidal activity of this species. The cytotoxicity activity of H. velutina fractions and compounds of crude ethanolic extract of the aerial parts of this species was assessed by using fluorescence microscopy and propidium iodide staining. In addition, the production of nitric oxide (NO) and hemocyte recruitment were checked after different periods of exposure. The fluorescence microscopy revealed an increasing in larvae cell necrosis for the dichloromethane fraction, 7,4'-di-O-methyl-8-O-sulphate flavone and hexane fraction (15.4, 11.0, and 7.0%, respectively). The tiliroside did not show necrotic cells, which showed the same result as that seen in the negative control. The NO concentration in hemolymph after 24 h exposure was significantly greater for the dichloromethane fraction and the 7,4'-di-O-methyl-8-O-sulphate flavone (123.8 and 56.2 µM, respectively) when compared to the hexane fraction and tiliroside (10.8 and 8.3 µM, respectively). The presence of plasmocytes only in the dichloromethane fraction and 7,4'-di-O-methyl-8-O-sulphate flavone treatments suggest that these would be the hemocytes responsible for the highest NO production, acting as a defense agent. Our results showed that the larvicidal activity developed by H. velutina compounds is related to its hemocyte necrotizing activity and alteration in NO production.

Aqueous extract from Luehea divaricata Mart. Leaves reduces nociception in rats with neuropathic pain.

ETHNOPHARMACOLOGICAL RELEVANCE: Luehea divaricata, popularly known in Brazil as "açoita-cavalo", has been widely explored by different ethnic groups native to Brazil to treat different pathologic conditions, including inflammatory pain. However, no report could be found on the effect that extract of L. divaricata has on neuropathic pain. This is an important topic because convergent and divergent mechanisms underlie inflammatory vs. neuropathic pain indicate that there may not always be a clear mechanistic delineation between these two conditions. AIM OF THE STUDY: The study aimed to determine antioxidant activity and macronutrient composition of aqueous extract from leaves of L. divaricata, and the effect of oral administration on nociception in rats with chronic constriction injury (CCI) of sciatic nerve-induced neuropathic pain, one of the most commonly employed animal models of neuropathic pain. MATERIALS AND METHODS: The antioxidant activity of the extract was evaluated by total phenolic content and DPPH, ABTS●+ and ORAC methods. Vitexin was determined by HPLC to show that the composition of the extract of the present study is similar to that used in previous studies with this genus. Total sugar and sucrose concentrations were assessed by the anthrone method, while glucose and triacilglycerides were determined using commercially available kits. Fructose concentration was calculated from values for total sugars, glucose and sucrose. Total protein was determined by Bradford assay. The effect on DNA strand breaking was investigated by inhibition of strand breaking of supercoiled DNA by hydroxyl radical. The antinociceptive effects of aqueous extract (100, 300, 500, and 1000 mg/kg, i.g.) were evaluated on thermal and mechanical thresholds for neuropathic pain induced by chronic constriction injury (CCI) of the sciatic nerve in rats. We also compared the antinociceptive effect of the extract (500 mg/kg, i.g.) with that induced by gabapentin (50 mg/kg, i.g.), a first-line clinical treatment for neuropathic pain. The effect of co-administration of extract (500 mg/kg, i.g.) and low-dose gabapentin (30 mg/kg, i.g.) was also assessed. In addition, the effect of the extract on body weight, and blood and hepatic parameters were investigated to reveal possible side effects of treatment. RESULTS: The extract showed high content of total phenol; good reducing capacity for DPPH, ABTS●+ and ORAC assays; presence of vitexin; and a high capacity to inhibit strand breaking of supercoiled DNA. The predominant sugar was sucrose, followed by glucose and fructose. Total protein was greater than triacylglycerides, with the latter being present in a trace amount in the extract. The extract increased the thermal and mechanical thresholds, which was reduced by CCI. The antinociceptive effect was comparable to gabapentin and was also found after co-administration of extract and low-dose gabapentin. No significant change was found in body weight and blood and hepatic indicators after extract treatment. CONCLUSIONS: Aqueous extract from L. divaricata leaves was as effective as gabapentin at attenuating CCI-induced neuropathic pain, indicating for first time the therapeutic potential of this species for this type of pain.

Anti-inflammatory and anti-nociceptive activities of essential oil of Waltheria indica / Actividades antiinflamatorias y nociceptivas del aceite esencial de Waltheria indica

This paper reports for the first time volatile compounds, anti-nociceptive and anti-inflammatory activities of essential oils from the leaves of Waltheria indica L. (Stericullaceae) growing in Nigeria. The essential oil was hydro-distilled and characterized by gas chromatography-flame ionization detection (GC-FID) and gas chromatography coupled with mass spectrometry (GC-MS) analyses. The anti-inflammatory activity was evaluated on carrageenan induced rat paw edema while the anti-nociceptive test was based on hot plate model. The hydro-distillation afforded 0.41% (dry weight basis) of light green oil. Forty compounds representing 99.8% were identified in the oil. The main constituents of the oil were limonene (34.7%), sabinene (21.2%) and citronellal (9.7%). The anti-nociceptive property of the essential oils statically inhibited edema development (p<0.001) at a dose of 200 and 400 mg/kg independent of time of exposure. However, the 100 mg/kg Waltheria indica essential oils (WIEO) displayed a relatively low inhibition (p<0.01-p>0.5) which declines as exposure time increases. The anti-inflammatory activities shows a steady rate and non-dose dependent activity (p<0.001) up to the 3rd h of inflammation study. Conversely, a sharp reduction at the rate of p<0.5, 0.1 and 0.01 for the 100, 200 and 400 mg/kg WIEO doses respectively. Overall, the results presented sustain and establish the anti-nociceptive and anti-inflammatory properties and justifies the need for further evaluation and development of the essential oils from this plant.

Este artículo informa por primera vez de compuestos volátiles, actividades anti-nociceptivas y antiinflamatorias de aceites esenciales de las hojas de Waltheria indica L. (Stericullaceae) que crecen en Nigeria. El aceite esencial fue hidro-destilado y se caracterizó por cromatografía de gases-detección de ionización de llama (GC-FID) y cromatografía de gases junto con análisis de espectrometría de masas (GC-MS). La actividad antiinflamatoria se evaluó en el edema de pata de rata inducido por carragenano, mientras que la prueba antinociceptiva se basó en el modelo de placa caliente. La destilación hidráulica proporcionó 0,41% (en peso seco) de aceite verde claro. Cuarenta compuestos que representan el 99.8% fueron identificados en el aceite. Los principales componentes del aceite fueron el limoneno (34,7%), el sabineno (21,2%) y el citronelal (9,7%). La propiedad anti-nociceptiva de los aceites esenciales inhibió estáticamente el desarrollo del edema (p<0.001) a una dosis de 200 y 400 mg/kg independientemente del tiempo de exposición. Sin embargo, los aceites esenciales de Waltheria indica de 100 mg/kg (WIEO) mostraron una inhibición relativamente baja (p<0.01-p>0.5) que disminuye a medida que aumenta el tiempo de exposición. Las actividades antiinflamatorias muestran una tasa constante y una actividad no dependiente de la dosis (p<0.001) hasta la tercera hora del estudio de inflamación. Por el contrario, una fuerte reducción a una tasa de p<0.5, 0.1 y 0.01 para las dosis de 100, 200 y 400 mg/kg de WIEO respectivamente. En general, los resultados presentados sostienen y establecen las propiedades anti-nociceptivas y antiinflamatorias y justifican la necesidad de una mayor evaluación y desarrollo de los aceites esenciales de esta planta.

Anti-inflammatory and cytotoxic activities of the extracts, fractions, and chemical constituents isolated from Luehea ochrophylla Mart.

BACKGROUND: Stem bark of Luehea ochrophylla (L. ochrophylla) is used by the traditional Brazilian medicine for treatment of rheumatic diseases and tumors. This study aimed to investigate inhibition of acute and chronic inflammations and cytotoxic activity of extracts, fractions, and isolated compounds from L. ochrophylla. METHODS: Hexane (HE) and ethanol (EE) extracts obtained from stem bark of L. ochrophylla were submitted to chromatographic fractionation. In order to test acute inflammation, experimental model of impact injury was used, followed by transdermal application of gels using phonophoresis. Histological analysis was based on scores assigned by the capacity of decreasing the lesion. To evaluate the effect EE and fractions on cell proliferation, human lymphocytes were stimulated with phytohemagglutinin and analyzed using flow cytometry. Proliferation was measured using VPD 450 staining and the calculated proliferative index (PI). The cytotoxic activity was evaluated using MTT colorimetric method against MDA-MB-231, MCF-7, HCT-116, and Vero cells. GraphPad Prism Version 5 was used for statistical analysis. RESULTS: HE and EE provided friedelin, ß-friedelinol, lupeol, mixture of lupeol and pseudotaraxasterol, ß-sitosterol, betulinic acid, mixture of lupeol and taraxasterol, (-)-epicatechin, ß-sitosterol-3-O-ß-D-glucopyranoside, and (+)-epicatechin-(4ß-8)-epicatechin. HE, ethyl acetate fraction (AF), betulinic acid, and ß-sitosterol promoted regeneration of muscle fibers caused by muscle injury. AF significantly (p < 0.05) reduced the lymphocyte proliferation index (1.36 for cultures stimulated with PHA, 0.7 for untreated cultures and 0.12 for cultures stimulated with PHA and treated with AF 25 µg/mL and AF 50 µg/mL, respectively). ß-Sitosterol-3-O-ß-D-glucopyranoside exhibited high cytotoxic activity (IC50 = 1.279 µg/mL) against HCT-116 cell line. CONCLUSION: These results suggest that extracts, fractions, and chemical constituents from L. ochrophylla decreases inflammatory processes generated by muscle injury. The anti-inflammatory activity may be justified by high inhibition of T cell proliferation. These extracts, fractions, and chemical constituents from L. ochrophylla may be useful as a therapeutic agent against rheumatic diseases. Moreover, chemical constituents from L. ochrophylla show potent cytotoxic activity against colon and rectal carcinomas.

Larvicidal Compounds Extracted from Helicteres velutina K. Schum (Sterculiaceae) Evaluated against Aedes aegypti L.

Helicteres velutina K. Schum (Sterculiaceae), a member of Malvaceae sensu lato, is a Brazilian endemic plant that has been used by the indigenous tribe Pankarare as an insect repellent. A previous study has reported the isolation of terpenoids, flavonoids and pheophytins, in addition to the larvicidal activity of crude H. velutina extracts derived from the aerial components (leaves, branches/twigs, and flowers). The present study reports the biomonitoring of the effects of fractions and isolated compounds derived from H. velutina against A. aegypti fourth instar larvae. A crude ethanol extract was submitted to liquid-liquid extraction with hexane, dichloromethane, ethyl acetate and n-butanol to obtain their respective fractions. Larvicidal evaluations of the fractions were performed, and the hexane and dichloromethane fractions exhibited greater activities than the other fractions, with LC50 (50% lethal concentration) values of 3.88 and 5.80 mg/mL, respectively. The phytochemical study of these fractions resulted in the isolation and identification of 17 compounds. The molecules were subjected to a virtual screening protocol, and five molecules presented potential larvicidal activity after analyses of their applicability domains. When molecular docking was analysed, only three of these compounds showed an ability to bind with sterol carrier protein-2 (1PZ4), a protein found in the larval intestine. The compounds tiliroside and 7,4'-di-O-methyl-8-O-sulphate flavone showed in vitro larvicidal activity, with LC50 values of 0.275 mg/mL after 72 h and 0.182 mg/mL after 24 h of exposure, respectively. This is the first study to demonstrate the larvicidal activity of sulphated flavonoids against A. aegypti. Our results showed that the presence of the OSO3H group attached to C-8 of the flavonoid was crucial to the larvicidal activity. This research supports the traditional use of H. velutina as an alternative insecticide for the control of A. aegypti, which is a vector for severe arboviruses, such as dengue and chikungunya.

Influence of Luehea divaricata Mart. extracts on peripheral vascular resistance and the role of nitric oxide and both Ca+2-sensitive and Kir6.1 ATP-sensitive K+ channels in the vasodilatory effects of isovitexin on isolated perfused mesenteric beds.

BACKGROUND: Luehea divaricata Mart. (Malvaceae) is an important medicinal species widely used by indigenous and riverside populations of the Brazilian Pantanal region. It has been shown that the several extracts obtained from leaves of this species have important cardioprotective effects. Nevertheless, the secondary metabolites responsible for this activity, as well as the molecular mechanisms responsible for their pharmacological effects remain unknown. PURPOSE: To carry out a biomonitoring study to identify possible active metabolites present in different ESLD fractions and evaluate the mechanisms responsible for the vasodilatory effects on isolated perfused mesenteric beds. METHODS: First, ESLD was obtained from L. divaricata leaves and a liquid-liquid fractionation was performed. The resulting fractions were analyzed by liquid chromatography-mass spectrometry. Then, the possible vasodilatory effects of ESLD, chloroform, ethyl acetate, n-butanolic and aqueous fractions on perfused arterial mesenteric vascular beds were evaluated. Finally, the molecular mechanisms involved in vasodilator responses of the aqueous fraction and its chemical component, isovitexin, on the mesenteric arteriolar tone were also investigated. RESULTS: In preparations with functional endothelium ESLD, n-butanolic, aqueous fraction and isovitexin dose-dependently reduced the perfusion pressure in mesenteric vascular beds. Endothelium removal or inhibition of nitric oxide synthase enzymes by L-NAME reduced the vasodilatory effects induced by aqueous fraction and isovitexin. Perfusion with nutritive solution containing 40 mM KCl abolished the vasodilatory effect of all aqueous fractions and Isovitexin doses. Treatment with glibenclamide, a Kir6.1 (ATP-sensitive) potassium channels blocker, tetraethylammonium, a non-selective KCa (calcium-activated) potassium channels blocker, or apamin, a potent blocker of small conductance Ca2+-activated (SK KCa) potassium channels reduced by around 70% vasodilation induced by all aqueous fractions and isovitexin doses. In addition, association of tetraethylammonium and glibenclamide, or L-NAME and glibenclamide, fully inhibited aqueous fraction and Isovitexin -induced vasodilation. CONCLUSION: This study showed that AqueFr obtained from Luehea divaricata and its metabolite - isovitexin - has important vasodilatory effects on MVBs. Apparently, these effects are dependent on endothelium-NO release and both SK KCa K+ channels and Kir6.1 ATP-sensitive K+ channels activation in the vascular smooth muscle.

In vitro activities of Ceiba speciosa (A.St.-Hil) Ravenna aqueous stem bark extract.

Several species of the genus Ceiba (Malvaceae) are ethnopharmacologically used. Thus, this study aimed to investigate the in vitro beneficial properties of the aqueous stem bark extract of Ceiba speciosa. The extract presented a great amount of phenolic compounds (117.4 ± 6.2 mg GAE/g). The antioxidant activity was assessed by DPPH (IC50 = 42.87 µg/mL), ORAC (2351.17 µmol TE/g) and FRAP (235.94 µM FeSO4/g) methods. In addition, the extract reduced MCF-7 cell viability as assessed by MTT. However, it prevented mitochondrial membrane depolarization and reduced caspase-9 activity induced by hydrogen peroxide. In conclusion, these findings indicate the extract is an excellent source of natural antioxidants and is able to protect ROS-induced cell death. Therefore, C. speciosa extract may possess beneficial properties for application in pharmaceutical industry as an antioxidant. However, further studies to better elucidate its mechanisms and to isolate its active compounds are required.

Sida tuberculata extract reduces the nociceptive response by chemical noxious stimuli in mice: Implications for mechanism of action, relation to chemical composition and molecular docking.

Sida tuberculata R.E.Fr. (Malvaceae) is a medicinal plant widely found in Southern Brazil, and popularly used for inflammatory disorders and to pain relief. A phytochemical analysis followed by an investigation about antinociceptive potential and mechanism of action were performed with leaves and roots extracts. Methanolic extracts, designated as S. tuberculata leaves extract (STLE) and S. tuberculata roots extract, were analyzed both by UHPLC­MS. The in vivo antinociceptive potential of STLE (10­300 mg kg−1) was assessed in mice subjected to the acetic acid­induced abdominal writhes and formalin model. Agonist/antagonist tests and computational docking suggest the involvement of opioid and adenosinergic systems. The main chemical class detected on extracts was the ecdysteroids, and 20­hydoxyecdysone (20HE) was confirmed as the major phytoconstituent. The pretreatment with STLE (100 mg kg−1) reduced more than 70% abdominal contortions induced by acetic acid model and produced significant inhibition on formalin­induced licking response. The mechanism of action study revealed STLE might act through opioid and adenosine systems. Molecular docking suggested kaempferol derivative and 20HE might interacting with µ­opioid receptor. Thus, the results suggest the existence of antinociceptive potential from S. tuberculata extracts being in accordance to the traditional use.

New Sulphated Flavonoids and Larvicidal Activity of Helicteres velutina K. Schum (Sterculiaceae).

Helicteres velutina K. Schum (Sterculiaceae), commonly known in Brazil as 'pitó', is traditionally used by indigenous peoples as insecticides and repellents. The present work reports on the the phytoconstituents from aerial parts of H. velutina and evaluation of the larvicidal potential of its extract. The compounds were isolated using chromatographic techniques and identified by NMR, IR and LC-HRMS. This study led to the isolation of a fatty acid, one aliphatic alcohol, four chlorophyll derivatives, one steroid, triterpenes, a lignan, and flavonoids, highlighting the new compounds in the literature, 5,4'-di-hydroxy-7-methoxy-8-O-sulphate flavone (mariahine) (15a) and 5,3'-di-hydroxy-7,4'-dimethoxy-8-O-sulphate flavone (condadine) (15b). The work presented here contributes to the chemotaxonomic knowledge of the Sterculiaceae family by describing the occurrence of sulphated flavonoids in this family for the first time. The crude ethanolic extract of H. velutina featured robust larvicidal activity against Aedes aegypti larvae, showing that the extract can be useful as a domestic larvicide, just as indicated by traditional use, to combat A. aegypti, a vector insect of severe viral diseases, such as dengue and Zika.

Guazuma ulmifolia Lam. Decreases Oxidative Stress in Blood Cells and Prevents Doxorubicin-Induced Cardiotoxicity.

Doxorubicin (DOX) is an efficient chemotherapeutic agent, but its clinical application is limited by its cardiotoxicity associated with increased oxidative stress. Thus, the combination of DOX and antioxidants has been encouraged. In this study, we evaluated (I) the chemical composition and antioxidant capacity of aqueous extracts from Guazuma ulmifolia stem bark (GUEsb) and leaves (GUEl) in 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging, 2,2'-azobis(2-amidinopropane) dihydrochloride- (AAPH-) or DOX-induced lipid peroxidation inhibition in human blood cells, and intracellular reactive oxygen species (ROS) quantification using the fluorescent probe dichloro-dihydro-fluorescein diacetate (DCFH-DA) in K562 erythroleukemia cells incubated with GUEsb and stimulated with hydrogen peroxide; (II) the viability of K562 cells and human leukocytes treated with GUEsb in the absence or presence of DOX using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; (III) the acute toxicity of GUEsb; and (IV) the cardioprotective effect of GUEsb in C57Bl/6 mice treated with DOX. The chemical composition indicated the presence of flavan-3-ol derivatives and condensed tannins in GUEsb and glycosylated flavonoids in GUEl. GUEsb and GUEl showed free-radical scavenging antioxidant activity, antihemolytic activity, and AAPH- as well as DOX-induced malondialdehyde content reduction in human erythrocytes. Based on its higher antioxidant potential, GUEsb was selected and subsequently showed intracellular ROS reduction without impairing the chemotherapeutic activity of DOX in K562 cells or inducing leukocyte cell death, but protected them against DOX-induced cell death. Yet, GUEsb did not show in vivo acute toxicity, and it prevented MDA generation in the cardiac tissue of DOX-treated mice, thus demonstrating its cardioprotective effect. Taken together, the results show that GUEsb and GUEl are natural alternatives to treat diseases associated with oxidative stress and that, in particular, GUEsb may play an adjuvant role in DOX chemotherapy.

Anti-Helicobacter pylori Potential of Three Edible Plants Known as Quelites in Mexico.

"Quelites" are edible plants that are part of the traditional agro-ecosystems in Mexico. These plants, despite their already known nutritional properties, are now considered neglected and underutilized species. With the objective of promoting their reinsertion in the markets and mainly, in daily diets, efforts have been made to study them from multidisciplinary approaches to demonstrate their beneficial properties. To generate evidence of an added health-promoting value that would encourage quelites consumption, in the present work, the anti-Helicobacter pylori activity of three representative quelite species, Anoda cristata (Alache), Cnidoscolus aconitifolius (Chaya), and Crotalaria pumila (Chepil), was tested. H. pylori is considered the etiological agent of gastritis, ulcer, and gastric cancer, and represents a public health problem in Mexico and worldwide. Aqueous (AQ) and dichloromethane-methanol (DM) extracts were obtained from the three species of quelites to investigate their effect on H. pylori growth and on two of its colonization factors (adherence and urease activity). DM extracts from Chaya, Chepil, and Alache exert the best inhibitory effect on bacterial growth, with minimum inhibitory concentrations of 62.5, 125, and 250 µg/mL, respectively. AQ and DM extracts inhibit bacterial adhesion by 30% to 50%. None of them has an effect on urease activity. The two flavonoids present in A. cristata, acacetin and diosmetin, inhibit H. pylori growth by ∼90% with 3.9 µg/mL. These results provide new information about the anti-H. pylori potential of three edible quelites, and give an added value, since their routine consumption may impact on the prevention and/or control of H. pylori-associated diseases.

Neuropharmacological and neuroprotective activities of some metabolites produced by cell suspension culture of Waltheria americana Linn.

Waltheria americana is a plant used in Mexican traditional medicine to treat some nervous system disorders. The aims of the present study were to isolate and determine the neuropharmacological and neurprotective activities of metabolites produced by a cell suspension culture of Waltheria americana. Submerged cultivation of W. americana cells provided biomass. A methanol-soluble extract (WAsc) was obtained from biomass. WAsc was fractionated yielding the chromatographic fractions 4WAsc-H2O and WAsc-CH2Cl2. For the determination of anticonvulsant activity in vivo, seizures were induced in mice by pentylenetetrazol (PTZ). Neuropharmacological activities (release of gamma amino butyric acid (GABA) and neuroprotection) of chromatographic fractions were determined by in vitro histological analysis of brain sections of mice post mortem. Fraction 4WAsc-H2O (containing saccharides) did not produce neuronal damage, neurodegeneration, interstitial tissue edema, astrocytic activation, nor cell death. Pretreatment of animals with 4WAsc-H2O and WAsc-CH2Cl2 from W. americana cell suspensions induced an increase in: GABA release, seizure latency, survival time, neuroprotection, and a decrease in the degree of severity of tonic/tonic-clonic convulsions, preventing PTZ-induced death of up to 100% of animals of study. Bioactive compounds produced in suspension cell culture of W. americana produce neuroprotective and neuropharmacological activities associated with the GABAergic neurotransmission system.

Níveis de inclusão da torta da semente de cupuaçu na dieta de frangos caipiras criados na amazônia ocidental / Levels of cupuaçu seed by-product in feedstuff for free-range broilers reared in the western amazon / Niveles de inclusión del subproducto de semilla de cupuaçu en la dieta de pollos campesinos creados en la amazónia ocidental

O experimento foi realizado com o intuito de avaliar os efeitos da inclusão da torta da semente de cupuaçu (TSC) nas dietas de frangos de linhagem caipira. O delineamento foi o inteiramente casualizado e foram utilizados 300 pintainhos de corte, sendo que os níveis de inclusão foram de 0, 5, 10, 15 e 20%, de TSC nas rações. Cada tratamento possuía seis repetições com 10 aves. Foram analisados os níveis de inclusões da TSC nos períodos de 1-14, 1-28, 1-42, 1-56 e 1-70 dias de criação, em relação ao desempenho zootécnico (consumo de ração, peso vivo, conversão alimentar, eficiência alimentar e viabilidade dos frangos). Foi analisado também o rendimento de carcaça e a margem bruta relativa (MBR). A utilização da TSC na dieta dos frangos reduziu o consumo de ração e, consequentemente, o peso vivo, piorando a conversão alimentar, reduzindo a eficiência alimentar da ração e a viabilidade dos frangos. Não houve influência da inclusão da TSC sobre o rendimento de carcaça de machos, porém houve aumento no rendimento de moela vazia, intestinos e redução da gordura abdominal. Houve também redução na MBR, conforme os níveis crescentes de inclusão da TSC.(AU)

The experiment aimed to evaluate the effects of including the by-product of cupuacu seeds (BCS) in dietary feed of free-range broilers. A completely randomized design was used with 300 broiler chickens, and the inclusion levels ranged from 0, 5, 10, 15 to 20% of BCS in the feeds. Each treatment had six replicates with 10 birds. The BCS inclusion levels were analyzed in the total period from 1-14, 1-28, 1-42, 1-56, and 1-70 days old, in relation to the production performance (feed intake, live weight, feed conversion, feed efficiency and viability of broilers). The carcass yield and gross margin ratio (GMR) were also analyzed. The use of BCS in the feed of broiler chickens reduced the feed intake and consequently the live weight, worsening the feed conversion, and also reduced feed efficiency and the viability of broiler chickens. There was no influence of the inclusion levels of BCS on the carcass yield of males; nevertheless, there was an increase in the yield of intestines and gizzard, and a reduction of abdominal fat. There was also a reduction in the GMR as the BCS levels increased.(AU)

El experimento fue realizado con el objetivo de evaluar los efectos de la inclusión del subproducto de semilla de cupuaçu (SSC) en las dietas de pollos de linaje campesino. El delineamiento fue enteramente casual y fueron utilizados 300 pollitos, siendo que los niveles de inclusión fueron de 0,5, 10, 15 y 20% de SSC en las raciones. Cada tratamiento poseía seis repeticiones con 10 aves. Se analizó los niveles de inclusiones del SSC en los períodos de 1-14, 1-28, 1-42, 1-56 y 1-70 días de creación, en relación al desempeño zootécnico (consumo de ración, peso vivo, conversión alimentar, eficiencia alimentar y viabilidad de los pollos). Se ha analizado también el rendimiento del esqueleto y la margen bruta relativa (MBR). La utilización del SSC en la dieta de los pollos acortó el consumo de ración y consecuentemente el peso vivo, afectando de forma negativa la conversión alimentar, reduciendo la eficiencia alimentar de la ración y la viabilidad de los pollos. No hubo influencia de la inclusión del SSC sobre el rendimiento del esqueleto de machos, pero hubo aumento en el rendimiento de molleja vacía, intestinos y reducción de grasa abdominal. Hubo también reducción en la MBR, conforme los niveles crecientes de inclusión del SSC.(AU)

Chemical composition and biological activities of Helicteres vegae and Heliopsis sinaloensis.

CONTEXT: Helicteres vegae Cristóbal (Sterculiaceae) (Hv) and Heliopsis sinaloensis B.L. Turner (Asteraceae) (Hs) are endangered and poorly studied plant species; related plants have been used against chronic-degenerative and infectious diseases. Therefore, Hv and Hs could be sources of bioactive compounds against these illnesses. OBJECTIVE: To determine the chemical composition and biological activities (antioxidant, antimutagenic and antimicrobial) of Hv and Hs leaves (L) and stems (S). MATERIALS AND METHODS: Methanol extracts (ME) of each plant/tissue were evaluated for their phytochemicals; phenolics (HPLC-DAD-ESI-MS); antioxidant activity (AA) (0.125-4 mg/mL) (DPPH, ABTS, ORAC and ß-carotene discoloration); antimutagenicity (0.5 and 1 mg/plate) (Ames assay, tester strain Salmonella enterica serovar Typhimurium YG1024, 1-nitropyrene as mutagen); activity against human pathogens (1 mg/mL); and toxicity (0.01-2 mg/mL) (Artemia salina assay). RESULTS: All ME showed flavonoids and triterpenes/steroids. The ME-SHv had the highest content of total phenolics (TP) (2245.82 ± 21.45 mg GAE/100 g d.w.) and condensed tannins (603.71 ± 1.115 mg CE/100 g d.w.). The compounds identified were flavonoids (kaempferol 7-O-coumaroylhexoside, and two kaempferol 7-O-rhamnosylhexosides) and phenolics [rosmarinic acid, and 3'-O-(8″-Z-caffeoyl) rosmarinic acid]. The ME-LHs showed the highest content of flavonoids (357.88 mg RE/g d.w.) and phenolic acids (238.58 mg CAE/g d.w.) by HPLC. The ME-SHv showed the highest AA. All ME were strong antimutagens (63.3-85.7%). Only the Hs extracts were toxic (ME-LHs, LC50 = 94.9 ± 1.7 µg/mL; ME-SHs, LC50 = 89.03 ± 4.42 µg/mL). DISCUSSION AND CONCLUSIONS: Both Hv and Hs are potential sources of preventive and therapeutic agents against chronic-degenerative diseases.

Swainsonine-induced lysosomal storage disease in goats caused by the ingestion of Sida rodrigoi Monteiro in North-western Argentina.

There are numerous poisonous plants that can induce intralysosomal accumulation of glycoproteins and neurologic syndromes. Here we describe for the first time, a disease caused by ingesting Sida rodrigoi Monteiro in goats in North-western Argentina. The animals showed weight loss, indifference to the environment, unsteady gait and ataxia. Histopathologic studies showed vacuolization in cells of various organs, mainly in the CNS. The material deposited in the cells was positive for LCA (Lens culinaris agglutinin), WGA (Triticum vulgaris agglutinin), sWGA (succinyl-Triticum vulgaris agglutinin) and Con-A (Concanavalia ensiformis agglutinin) lectins. Finally, toxic levels of swansonine were identified in the plant. The present investigation allowed to recognize S. rodrigoi Monteiro poisoning as a plant induced α-mannosidosis.

Alkaloids and Phenolic Compounds from Sida rhombifolia L. (Malvaceae) and Vasorelaxant Activity of Two Indoquinoline Alkaloids.

The follow-up of phytochemical and pharmacological studies of Sida rhombifolia L. (Malvaceae) aims to strengthen the chemosystematics and pharmacology of Sida genera and support the ethnopharmacological use of this species as hypotensive herb. The present work reports phytoconstituents isolated and identified from aerial parts of S. rhombifolia by using chromatographic and spectroscopic methods. The study led to the isolation of scopoletin (1), scoporone (2), ethoxy-ferulate (3), kaempferol (4), kaempferol-3-O-ß-d-glycosyl-6''-α-d-rhamnose (5), quindolinone (6), 11-methoxy-quindoline (7), quindoline (8), and the cryptolepine salt (9). The alkaloids quindolinone (6) and cryptolepine salt (9) showed vasorelaxant activity in rodent isolated mesenteric arteries.