Larvicidal and Adulticidal Activity of Essential Oils from Four Cuban Plants against Three Mosquito Vector Species.

Mosquitoes are one of the main vectors of many important diseases and their degree of resistance to chemical insecticides has increased. Nowadays, it has become crucial to identify novel plant larvicides with an eco-friendly impact. The components of essential oils from Croton linearis Jacq. (EO-Cl), Lantana involucrata L. (EO-Li), Ocimum sanctum var. cubensis M. Gómez. (EO-Os), and Zanthoxylum pistaciifolium Griseb. (syn. Zanthoxylum flavum subsp. pistaciifolium (Griseb.) Reynel (EO-Zp) were determined using gas chromatography-mass spectrometry (GC-MS) analysis. Larvicidal and adulticidal bioassays against Aedes aegypti, Anopheles albitarsis and Culex quinquefasciatus, were performed according to the World Health Organization standard methods. A high diversity of compounds was identified in the four oils, with a total of 152 compounds (33-70 components). EO-Cl, EO-Li, and EO-Os were classified as active against both insect forms, larvae and adults. Lantana involucrata showed the best results, with LC50 values from 33.8 to 41.7 mg/L. In most of the cases, it was not possible to associate the main compounds with the measured activity, supporting the hypothesis about probable synergistic interactions among major and minor compounds. The results indicate EO-Cl, EO-Os, and EO-Li as good eco-friendly insecticides with potential.

Chemical Composition and In Vitro and In Silico Antileishmanial Evaluation of the Essential Oil from Croton linearis Jacq. Stems.

Croton linearis Jacq. is an aromatic shrub that has been utilized in traditional medicine in the Bahamas, Jamaica, and Cuba. Recent studies have revealed the antiprotozoal potential of its leaves. The present work is aimed to identify the volatile constituents of essential oil from the stems of C. linearis (CLS-EO) and evaluate its in vitro antileishmanial activity. In addition, an in silico study of the molecular interactions was performed using molecular docking. A gas chromatographic-mass spectrometric analysis of CLS-EO identified 1,8-cineole (27.8%), α-pinene (11.1%), cis-sabinene (8.1%), p-cymene (5.7%), α-terpineol (4.4%), epi-γ-eudesmol (4.2%), linalool (3.9%), and terpinen-4-ol (2.6%) as major constituents. The evaluation of antileishmanial activity showed that CLS-EO has good activity on both parasite forms (IC50Promastigote = 21.4 ± 0.1 µg/mL; IC50Amastigote = 18.9 ± 0.3 µg/mL), with a CC50 of 49.0 ± 5.0 µg/mL on peritoneal macrophages from BALB/c mice (selectivity index = 2 and 3 using the promastigote and amastigote results). Molecular docking showed good binding of epi-γ-eudesmol with different target enzymes of Leishmania. This study is the first report of the chemical composition and anti-Leishmania evaluation of CLS-EO. These findings provide support for further studies of the antileishmanial effect of this product.

Inhibitory Effect on Nitric Oxide Release in LPS-Stimulated Macrophages and Free Radical Scavenging Activity of Croton linearis Jacq. Leaves.

Oxidative stress is an important component of many diseases including cancer, along with inflammatory and neurodegenerative processes. Natural antioxidants have emerged as promising substances to protect the human body against reactive oxygen and nitrogen species. The present study evaluates the inhibition of nitric oxide (NO) production in LPS-stimulated RAW 264.7 murine macrophages and the free radical scavenging activity of Croton linearis Jacq. leaves. UPLC-QTOF-MS analysis identified 18 compounds: nine alkaloids with a morphinane, benzylisoquinoline or aporphine nucleus, and nine O-glycosylated-flavonoids with quercetin, kaempferol and isorhamnetin as the aglycones. The crude extract (IC50 21.59 µg/mL) and the n-hexane fraction (IC50 4.88 µg/mL) significantly reduced the NO production in LPS-stimulated macrophages but with relatively high cytotoxicity (CC50 75.30 and CC50 70.12 µg/mL, respectively), while the ethyl acetate fraction also showed good activity (IC50 40.03 µg/mL) without affecting the RAW 264.7 cell viability. On the other hand, the crude extract, as well as the dichloromethane and ethyl acetate fractions, showed better DPPH and ABTS free radical scavenging activities. Considering the chemical composition and the activity observed for Croton linearis leaves, they may be considered a good source of antioxidants to combat oxidative damage-related diseases.

Inhibition of Bacterial Adhesion and Biofilm Formation by Seed-Derived Ethanol Extracts from Persea americana Mill.

The increase in antibiotic resistance demands innovative strategies to combat microorganisms. The current study evaluated the antibacterial and antivirulence effects of ethanol extracts from Persea americana seeds obtained by the Soxhlet (SE) and maceration (MaE) methods. The UHPLC-DAD-QTOF analysis showed mainly the presence of polyphenols and neolignan. Ethanol extracts were not cytotoxic to mammalian cells (CC50 > 500 µg/mL) and displayed a moderate antibacterial activity against Pseudomonas aeruginosa (IC50 = 87 and 187 µg/mL) and Staphylococcus aureus (IC50 = 144 and 159 µg/mL). Interestingly, no antibacterial activity was found against Escherichia coli. SE and MaE extracts were also able to significantly reduce the bacterial adhesion to A549 lung epithelial cells. Additionally, both extracts inhibited the biofilm growth at 24 h and facilitated the release of internal cell components in P. aeruginosa, which might be associated with cell membrane destabilization. Real-time PCR and agarose electrophoresis gel analysis indicated that avocado seed ethanol extracts (64 µg/mL) downregulated virulence-related factors such as mexT and lasA genes. Our results support the potential of bioproducts from P. americana seeds as anti-adhesive and anti-biofilm agents.

Potencial antioxidante del extracto de hojas de Spondias mombin L. en etanol al 70 % / Antioxidant potential of Spondias mombin L. leaf extract in 70% ethanol / Potencial antioxidante do extrato da folha de Spondias mombin L. em etanol 70%

RESUMEN Introducción: La especie Spondias mombin L. es una planta que crece en Cuba y a la que se le reporta un número considerables de aplicaciones etnofarmacológicas y elevado poder antioxidante. Sin embargo, la mayor parte de estos reportes está orientada al fruto, el cual resalta por su valor nutricional. Objetivo: Evaluar la actividad antioxidante de un extracto de hojas de Spondias mombin L. en etanol al 70 %. Método: La actividad antioxidante fue evaluada por seis métodos diferentes: determinación del índice de oxidación, poder reductor, inhibición de los radicales DPPH y ABTS, capacidad reductora del peróxido de hidrógeno y capacidad de neutralización del radical oxígeno. Se evaluaron concentraciones desde los 1 000 hasta los 62,5 µg/mL preparadas a partir del valor de sólidos totales determinados. Resultados: Para el índice de oxidación, concentraciones de 250 µg/mL resultaron suficientes para decolorar la solución de permanganato en igual período de tiempo que el patrón de quercetina. Sin embargo, los IC50 calculados para el poder reductor, la inhibición del radical DPPH y del ABTS resultaron mayores que el estándar de ácido ascórbico con valores de 362,33; 190,34 y 241,88 µg/mL, respectivamente. Conclusiones: El extracto de hojas de Spondias mombin L. en etanol al 70 % presenta una elevada acción antioxidante en los seis modelos experimentales ensayados, aun cuando en la mayor parte de casos, con valores inferiores al de la sustancia de referencia empleada. Estos resultados conjuntamente con la diversidad estructural de metabolitos presentes en el mismo resaltan su valor como potencial antioxidante natural.

ABSTRACT Introduction: The species Spondias mombin L. is a plant that grows in Cuba, and to which a considerable number of ethnopharmacological applications and high antioxidant power are reported. However, most of these reports are oriented to the fruit, which stands out for its nutritional value. Objective: To evaluate the antioxidant activity of an extract of Spondias mombin L. leaves in 70% ethanol. Method: The antioxidant activity was evaluated by six different methods: determination of the oxidation index, reducing power, inhibition of DPPH and ABTS radicals, hydrogen peroxide reducing capacity and oxygen radical neutralization capacity. Concentrations from 1000 to 62.5 µg/mL were evaluated, prepared from the value of total solids determined. Results: For the oxidation index, concentrations of 250 µg/mL were sufficient to decolorize the permanganate solution in the same period of time as the quercetin standard. However, the IC50 calculated for the reducing power, the inhibition of the DPPH radical and ABTS were higher than the ascorbic acid standard with values of 362.33; 190.34 and 241.88 µg/mL, respectively. Conclusions: The extract of leaves of Spondias mombin L. in 70% ethanol presents a high antioxidant action in the six experimental models tested, even though in most cases, with lower values than the reference substance used. These results, together with the structural diversity of metabolites present in it, highlight its value as a potential natural antioxidant.

RESUMO Introdução: A espécie Spondias mombin L. é uma planta que cresce em Cuba e à qual são relatados um número considerável de aplicações etnofarmacológicas e alto poder antioxidante. No entanto, a maioria desses relatos é voltada para o fruto, que se destaca pelo seu valor nutricional. Objetivo: Avaliar a atividade antioxidante de um extrato de folhas de Spondias mombin L. em etanol 70%. Método: A atividade antioxidante foi avaliada por seis diferentes métodos: determinação do índice de oxidação, poder redutor, inibição dos radicais DPPH e ABTS, capacidade redutora do peróxido de hidrogênio e capacidade de neutralização do radical oxigênio. Foram avaliadas concentrações de 1000 a 62,5 µg/mL, preparadas a partir do valor de sólidos totais determinado. Resultados: Para o índice de oxidação, concentrações de 250 µg/mL foram suficientes para descolorir a solução de permanganato no mesmo período de tempo do padrão de quercetina. No entanto, o IC50 calculado para o poder redutor, a inibição do radical DPPH e ABTS foram superiores ao padrão de ácido ascórbico com valores de 362,33; 190,34 e 241,88 µg/mL, respectivamente. Conclusões: O extrato de folhas de Spondias mombin L. em etanol 70% apresenta alta ação antioxidante nos seis modelos experimentais testados, embora na maioria dos casos com valores inferiores à substância de referência utilizada. Esses resultados, juntamente com a diversidade estrutural de metabólitos presentes nele, destacam seu valor como potencial antioxidante natural.

Antiplasmodial activity of alkaloids from Croton linearis leaves.

Croton linearis is a shrub that grows in Caribbean regions, which is rich in metabolites such as alkaloids. The main aim of this study was to evaluate the antiplasmodial effect of alkaloids from this species. Three isoquinoline alkaloids, i.e. reticuline (1), laudanidine (2) and 8,14-dihydrosalutaridine (3), were isolated from the leaves of C. linearis by flash chromatography and semi-preparative HPLC-DAD-MS. Their structures were elucidated by spectroscopic techniques. Antiplasmodial activity against the chloroquine-resistant strain Plasmodium falciparum K1 and cytotoxicity against MRC-5 cells (human fetal lung fibroblast cells) were assessed in vitro. Reticuline, laudanidine and 8,14-dihydrosalutaridine showed moderate antiplasmodial activity with IC50 values of 46.8 ± 0.6, 17.7 ± 0.6 and 16.0 ± 0.5 µM, respectively, but no cytotoxicity was observed in a concentration up to 64.0 µM. This is the first report on the antiplasmodial activity of laudanidine and 8,14-dihydrosalutaridine.

LC-MS Characterization and Biological Activities of Cuban Cultivars of Plectranthus neochilus Schltr.

Plectranthus neochilus Schltr. (Lamiaceae) is a plant recently introduced in Cuba. Worldwide, it is an ethnomedicinal alternative for its use against microbial infections, but the Cuban population use the extracts to treat sleep disorders. To address this apparent incongruity, four collections (from different seasonal conditions in the year) of Cuban P. neochilus cultivars were analyzed in terms of their pharmacognostic characteristics. Three extracts using fresh and dried leaves were chemically and biologically characterized. UPLC-DAD-MS/MS analysis was performed to determine their chemical composition, while a panel of nine microorganisms was used to evaluate their antimicrobial activity. Finally, cytotoxic effects of different fractions were measured in three cell lines by the resazurin viability assay. In contrast to previously reported micro and macromorphological properties of P. neochilus, the leaves from the Cuban cultivars did not present glandular trichomes, nor did they produce quantifiable levels of essential oils. Moreover, aqueous extracts used by the population revealed no significant antimicrobial activity and were not cytotoxic. The three extracts showed a similar phytochemical composition, i.e., eight flavonoids, seven abietane diterpenes, and rosmarinic acid as the major constituent, most of them reported for the first time in this species. The low yield of essential oil, the absence of glandular trichomes, compounds with a high level of oxidation, and a moderate antimicrobial activity detected were the most distinctive pharmacognostic and biological characteristics of P. neochilus grown in Cuba. These aspects could explain its non-use as an antimicrobial.

Novel flavonol-3-O-methylethers from Zanthoxylum pistaciifolium Griseb. (Rutaceae).

Zanthoxylum pistaciifolium Griseb. is a tree endemic to Cuba, occasionally used in herbal medicine. Previously, the antitrypanosomal activity of a n-hexane-2-butanone extract of Z. pistaciifolium leaves and of its constituent skimmianine were published. In the current study a more thorough examination of the respective extract is performed, which led to the isolation and identification of three flavonoids, more specifically, the flavonol-3-O-methylethers kaempferol-3-O-methylether (1) and novel compounds kaempferol-3-O-methylether-5-O-ß-D-glucoside (2) and kaempferol-8-hydroxy-3,7-O-dimethylether-5-O-ß-D-glucoside (3). All compounds were screened for their antimicrobial and antiprotozoal activity and cytotoxicity towards MRC-5 SV2 cells. Compound 1 showed a moderate to weak activity against Trypanosoma cruzi (IC50 30.8 µM), T. brucei (IC50 15.4 µM) and Plasmodium falciparum (IC50 53.8 µM), but also showed cytotoxicity (CC50 19.0 µM). Compounds 2 and 3 did not display activity in any of the assays (IC50 and CC50 > 64 µM).[Formula: see text].

Antifungals from plants: a bibliometric analysis of the last two decades (2000-2020) / Antifúngicos de plantas: un análisis bibliométrico de las últimas dos décadas (2000-2020)

This study aimed to analyse the world scientific production on antifungals from plants indexed in the Scopus database (2000-2020), highlighting the Cuban scientific production on this topic as a case study. Scopus raw data based on antifungal and plants hits were cleaned using the OpenRefine tool to eliminate similar and duplicate results. Cleaned data was processed using VOSviewer software for keyword-based bibliometric analysis. The 5000 most common words from the Corpus of Contemporary American English were excluded. A co-occurrence analysis was performed taking into account only authors keywords and using the complete count method. 24 084 documents were identified, of which 85.8percent were original articles and 9.6percent were reviews. The most productive journals were Journals of Ethnopharmacology, Natural Product Research and Fitoterapia. The annual scientific production was uniform with a linear growth. The thematic structure according to the co-occurrence network indicates that the most frequent terms were essential oils, antioxidant, medicinal plants, Candida albicans and flavonoids. In the case of Cuba, of the 64 publications on the subject, 90.6percent were original articles and 6.3percent were reviews. The Cuban magazines with the highest production were Revista Cubana de Plantas Medicinales and Biotecnología Aplicada. The most frequent terms according to the co-occurrence network in Cuban publications were Staphylococcus aureus, Candida albicans, plant leaf, antimicrobial activity, medicinal plants and flavonoids. These results prove that the global scientific community as well as the Cuban scientist are interested on the topic, further demonstrating that plants could become a reliable source of antifungal drugs(AU)

El objetivo del presente estudio fue analizar la producción científica mundial sobre antifúngicos de plantas recogida en la base de datos Scopus (2000-2020), con atención a la producción científica cubana como caso/estudio. Los datos resultantes de la búsqueda por coincidencia de las palabras antifungal y plants se filtraron con la herramienta OpenRefine, que eliminó resultados similares y duplicados. Se empleó el programa VOSviewer para el análisis bibliométrico basado en palabras claves, que excluyó las 5000 más comunes del Corpus of Contemporary American English. Se realizó un análisis de coocurrencia que consideró las palabras clave definidas por los autores en cada artículo a través del método de conteo completo. Se identificaron 24 084 documentos, de los cuales un 85,8 por ciento resultaron artículos originales y un 9,6 por ciento, artículos de revisión. Las revistas más productivas fueron Journals of Ethnopharmacology, Natural Product Research y Fitoterapia. La producción científica anual fue uniforme, con un crecimiento lineal. La red de coocurrencia indicó que los términos más frecuentes fueron aceites esenciales, antioxidante, plantas medicinales, Candida albicans y flavonoides. En Cuba, de las 64 publicaciones sobre el tema, el 90,6 por ciento resultaron ser artículos originales y 6,3 por ciento, artículos de revisión. Las revistas cubanas con mayor producción fueron Revista Cubana de Plantas Medicinales y Biotecnología Aplicada. Los términos más frecuentes en las publicaciones cubanas fueron Staphylococcus aureus, Candida albicans, hojas de plantas, actividad antimicrobiana, plantas medicinales y flavonoides. Estos resultados demuestran que, tanto la comunidad científica mundial como la cubana, manifiestan creciente interés en el tema, lo cual evidencia el potencial de las plantas como fuente confiable de medicamentos antifúngicos(AU)

Antifungal Activity of Extracts, Fractions, and Constituents from Coccoloba cowellii Leaves.

Coccoloba cowellii Britton (Polygonaceae, order Caryophyllales) is an endemic and critically endangered plant species that only grows in the municipality of Camagüey, a province of Cuba. A preliminary investigation of its total methanolic extract led to the discovery of promising antifungal activity. In this study, a bioassay-guided fractionation allowed the isolation of quercetin and four methoxyflavonoids: 3-O-methylquercetin, myricetin 3,3',4'-trimethyl ether, 6-methoxymyricetin 3,4'-dimethyl ether, and 6-methoxymyricetin 3,3',4'-trimethyl ether. The leaf extract, fractions, and compounds were tested against various fungi and showed strong in vitro antifungal activity against Cryptococcus neoformans and various Candida spp. with no cytotoxicity (CC50 > 64.0 µg/mL) on MRC-5 SV2 cells, determined by a resazurin assay. A Candida albicans SC5314 antibiofilm assay indicated that the antifungal activity of C. cowellii extracts and constituents is mainly targeted to planktonic cells. The total methanolic extract showed higher and broader activity compared with the fractions and mixture of compounds.

Anti-inflammatory effect of Adelia ricinella L. aerial parts.

OBJECTIVE: To investigate the main chemical components and the anti-inflammatory activity of extracts of Adelia ricinella L. aerial parts. METHODS: Three extracts obtained by soxhlet extraction and ethanol/water mixtures were evaluated in their chemical composition by UPLC-DAD-MS/MS. The in vitro anti-inflammatory activity of the prepared extracts was assessed through three different assays: COX-1 and COX-2 enzymatic inhibition, cell-based COX assays on RAW264.7 macrophages (ATCC) measuring the COX-2 protein expression by Western blot and the measurement of the PGE2 concentration in the supernatants of the culture medium. Also was determinate the effect of the three extracts on the RAW 264.7 cell viability. KEY FINDINGS: Few differences in the phytochemical profile were found between the three prepared extracts, identifying a blend of thirteen flavonoids derived from luteolin and apigenin, with orientin as main constituent. Plant extracts (alcoholic and aqueous) did not affect the macrophage cell viability (IC50 > 256 µg/ml) and significantly reduced COX-1 and COX-2 enzyme activities. Additionally, COX-2 expression and PGE2 release were suppressed after 24 h of LPS stimulation and treatment with plant extracts (8-64 µg/ml). CONCLUSIONS: A. ricinella extracts showed the ability to reduce the inflammatory effect exerted by LPS in murine macrophages. However, further studies should confirm their anti-inflammatory activity.

Chemical and Pharmacological Potential of Coccoloba cowellii, an Endemic Endangered Plant from Cuba.

Coccoloba cowellii Britton (Polygonaceae) is an endemic and critically endangered plant that only grows in Camagüey, a province of Cuba. In this study, a total of 13 compounds were identified in a methanolic leaf extract, employing a dereplication of the UHPLC-HRMS data by means of feature-based molecular networking (FBMN) analysis in the Global Natural Products Social Molecular Network (GNPS), together with the interpretation of the MS/MS data and comparison with the literature. The major constituents were glucuronides and glycosides of myricetin and quercetin, as well as epichatechin-3-O-gallate, catechin, epicatechin and gallic acid, all of them being reported for the first time in C. cowellii leaves. The leaf extract was also tested against various microorganisms, and it showed a strong antifungal effect against Candida albicans ATCC B59630 (azole-resistant) (IC50 2.1 µg/mL) and Cryptococcus neoformans ATCC B66663 (IC50 4.1 µg/mL) with no cytotoxicity (CC50 > 64.0 µg/mL) on MRC-5 SV2 cells, determined by the resazurin assay. Additionally, the extract strongly inhibited COX-1 and COX-2 enzyme activity using a cell-free experiment in a dose-dependent manner, being significantly more active on COX-1 (IC50 4.9 µg/mL) than on COX-2 (IC50 10.4 µg/mL). The constituents identified as well as the pharmacological activities measured highlight the potential of C. cowellii leaves, increasing the interest in the implementation of conservation strategies for this species.

Ethnobotany of Plectranthus neochilus Schltr (Meprobamate) in Cuba / Estudio etnobotánico de Plectranthus neochilus Schltr (Meprobamato) en Cuba

Chloroform extract (CE) and fractions obtained from Aldama arenaria roots were evaluated for their in vitro antiproliferative activity against 10 human tumor cell lines [leukemia (K-562), breast (MCF-7), ovary expressing a multidrug-resistant phenotype (NCI/ADR-RES), melanoma (UACC-62), lung (NCI-H460), prostate (PC-3), colon (HT29), ovary (OVCAR-3), glioma (U251), and kidney (786-0)]. CE presented weak to moderate antiproliferative activity (mean log GI50 1.07), whereas fractions 3 and 4, enriched with pimaranetype diterpenes [ent-pimara-8(14),15-dien-19-oic acid and ent-8(14),15-pimaradien-3ß-ol], presented moderate to potent activity for most cell lines, with mean log GI50 of 0.62 and 0.59, respectively. The results showed promising in vitro antiproliferative action of the samples obtained from A. arenaria, with the best results for NCI/ADR-RES, HT29, and OVCAR-3, and TGI values ranging from 5.95 to 28.71 µg.ml -1, demonstrating that compounds of this class may be potential prototypes for the discovery of new therapeutic agents.

El extracto de cloroformo (CE) y las fracciones obtenidas de las raíces de Aldama arenaria fueron evaluadas por su actividad antiproliferativa in vitro contra 10 líneas celulares tumorales humanas [leucemia (K-562), mama (MCF-7), ovario que expresa un fenotipo resistente a múltiples fármacos (NCI/ADR-RES), melanoma (UACC-62), pulmón (NCI-H460), próstata (PC-3), colon (HT29), ovario (OVCAR-3), glioma (U251) y riñón (786-0)]. CE presentó actividad antiproliferativa débil a moderada (log GI50 promedio de 1.07), mientras que las fracciones 3 y 4, enriquecidas con diterpenos de tipo pimarane [ent-pimara-8 (14), ácido 15-dien-19-oico y ent-8 (14), 15-pimaradien-3ß-ol], presentaron actividad moderada a potente para la mayoría de las líneas celulares, con un log GI50 promedio de 0.62 y 0.59, respectivamente. Los resultados mostraron una prometedora acción antiproliferativa in vitro de las muestras obtenidas de A. arenaria, con los mejores resultados para NCI/ADR-RES, HT29 y OVCAR-3, y valores de TGI que van desde 5.95 a 28.71 µg.ml -1, lo que demuestra que los compuestos de esta clase pueden ser prototipos potenciales para el descubrimiento de nuevos agentes terapéuticos.

Efectos de la corriente eléctrica de baja intensidad en ratones sanos / Effects of low-intensity electric current on healthy mice

Introducción: El conocimiento de los efectos de la terapia electroquímica, en los organismos puede ser importante para el tratamiento de diferentes enfermedades. Objetiv:o Evaluar los efectos de la corriente eléctrica directa de baja intensidad en ratones sanos. Métodos: 80 ratones C57BL/6/Cenp sanos (40 machos y 40 hembras) fueron distribuidos aleatoriamente en ocho grupos experimentales (cuatro grupos controles y cuatro grupos tratados). La corriente eléctrica directa (10 mA durante 5 min) o tensión constante (10 V durante 5 min) fue aplicada a los ratones. Fueron realizadas observaciones clínicas, macroscópicas, microscópicas y mediciones de los parámetros hematológicos y bioquímicos de la sangre. Resultados: Se indujeron cambios reversibles en los parámetros hematológicos, bioquímicos de la sangre e histológicos estudiados en ratones C57BL/6/Cenp machos y hembras cuando se usó 10 mA o 10 V y diferentes arreglos de electrodos, siendo menos notable para el modo de tensión constante. Conclusiones: Las alteraciones inducidas en los ratones C57BL/6/Cenp sanos por la corriente eléctrica directa de baja intensidad son reversibles y el proceso inflamatorio sistémico es dominado por los linfocitos(AU)

Introduction: Knowledge about the effects of electrochemotherapy on organisms may be important for the treatment of various diseases. Objective: Evaluate the effects of low-intensity direct electric current on healthy mice. Methods: Eighty healthy C57BL/6/Cenp mice (40 male and 40 female) were randomly distributed in eight experimental groups (four control and four treated). Direct electric current (10 mA during 5 min) or constant voltage (10 V during 5 min) was applied to the mice. Clinical, macroscopic and microscopic observation was performed, and measurements were taken of hematological and biochemical parameters of the blood. Results: Reversible changes were induced in hematological, biochemical and histological parameters of the blood of male and female C57BL/6/Cenp mice when 10 mA or 10 V and various electrode arrays were used. These changes were less noticeable in the constant voltage mode. Conclusions: The alterations induced by low-intensity direct electric current in healthy C57BL/6/Cenp mice are reversible. The systemic inflammatory process is dominated by the lymphocytes(AU)

Antimicrobial activity of leaf extracts and isolated constituents of Croton linearis.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves of Croton linearis, known as "rosemary", are widely used in folk medicine in Caribbean countries to treat fever and colds (associated to infections). AIM OF THE STUDY: To evaluate the in vitro antimicrobial activity of extracts and fractions derived from C. linearis leaves. MATERIALS AND METHODS: Bioassay-guided fractionation and isolation of compounds from an ethanolic extract of C. linearis, using flash chromatography and semi-preparative HPLC-DAD-MS (High Performance Liquid Chromatography - Diode Array Detection - Mass Spectrometry). Isolated compounds were characterized by MS, 1D- and 2D-NMR (Nuclear Magnetic Resonance) spectroscopy. The microdilution method with resazurin, as well as direct counting with an optical microscope, were used to determine the in vitro antimicrobial activity against bacteria, fungi and parasites. Moreover, the cytotoxicity on human fetal lung fibroblasts (MRC-5) was evaluated. RESULTS: The total extract and chloroform fraction (ClF) showed high activity against protozoa with IC50 values ranging from 1 to 26 µg/mL, but also cytotoxicity on MRC-5 and PMM (Peritoneal Murine Macrophages). Seven compounds were isolated and characterized for first time in this species: the alkaloids laudanidine, laudanosine, reticuline, corydine, glaucine and cularine and the flavonoid glycoside isorhamnetin-3-O-(6″O-p-trans-coumaroyl)-ß-glucopyranoside. Reticuline showed a weak activity against L. infantum (IC50 148.0 ±â€¯1.2 µM), while the flavonoid was active against T. cruzi (IC50 35.6 ±â€¯2.3 µM). CONCLUSIONS: The results show the antiprotozoal potential of the extract and some isolated constituents, which supports the use of this species in Caribbean folk medicine.

An ethnobotanical survey of medicinal plants used by inhabitants of Holguín, Eastern Region, Cuba / Estudio etnobotánico de las plantas medicinales usadas por los habitantes de Holguín, Región Oriental, Cuba

The present study was aimed to archive the etnhnomedicinal knowledge of plants used by inhabitants of seven villages of Holguín, Eastern region, Cuba. The ethnomedicinal information was collected through interviews. The collected data were analyzed through use value (UV), informant consensus factor (Fic) and fidelity level (FL). A total of 195 species of plants distributed in 166 genera belonging to 70 families were identified for the treatment of 17 ailment categories. The most treated conditions were digestive and liver disorders. The most important species according to their use value were Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson (0.236) and Annona muricata L. (0.194). Cancer and tumors had the Fic value of 0.94. A total of 19 species has a highest FL of 100 percent. This was the first ethnobotanical survey conducted in Holguín region, which will contribute to preserve valuable information of medicinal plants that may otherwise be lost to future generations.

El presente estudio tuvo como objetivo registrar el conocimiento etnomedicinal de las plantas usadas por los pobladores en 7 comunidades de Holguín, Región Oriental, Cuba. La información fue recogida a través de entrevistas y analizada cuantitativamente mediante indicadores etnobotánicos: valor de uso (UV), factor del consenso de los informantes (Fic) e índice de fidelidad (FL). Fueron reportadas un total de 195 especies de plantas, distribuidas en 166 géneros y 70 familias, para el tratamiento de 17 categorías de usos. Las indicaciones más frecuentes fueron los problemas digestivos y del hígado. Las especies medicinales con mayor UV fueron Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson (0.236) y Annona muricata L. (0.194). Cáncer y tumores tuvieron el valor más alto de Fic (0.94). Solo 19 especies presentaron un valor de FL de 100 %. Este primer estudio contribuirá a preservar la información de las plantas medicinales y que esta no se pierda en las futuras generaciones.

Bioassay-guided In vitro Study of the Antimicrobial and Cytotoxic Properties of the Leaves from Excoecaria Lucida Sw.

BACKGROUND: Excoecaria lucida Sw. (Euphorbiaceae) is a plant conventionally used throughout the Caribbean in the treatment of infectious diseases. OBJECTIVE: To evaluate, using bioassay-guided fractionation, the in vitro cytotoxicity and antimicrobial activity of E. lucida leaves. MATERIALS AND METHODS: A 95% ethanol crude extract was dried and fractionated by solid-liquid separation in four phases (hexane, dichloromethane, ethyl acetate, and butanol). Antimicrobial activity (3 bacteria, 6 yeasts, and 2 fungi) was evaluated by the dilution method with resazurin (2048, 512, 128, 32, and 8 µg/mL). The cytotoxicity assays were evaluated in two cell lines: MRC-5 and RAW 264.7; calculating the selectivity index. Assays were performed for the total extract, the isolated compound with the highest yield, and the ethyl acetate and butanol phases. Isolated compounds were characterized by nuclear magnetic resonance and mass spectrometry techniques. RESULTS: Fractionation process led to the isolation of ellagic acid (784.29 mg), 3,3',4'-tri-O-methyl ellagic 4-O-ß-D-glucopyranoside acid (6.1 mg), and corilagin (6.91 mg). The most active were ethyl acetate phase and ellagic acid with IC50= 128 µg/mL against seven and five different species of microorganisms, respectively. The total extract (IC50=512 µg/mL) and the ethyl acetate phase (IC50=128 µg/mL) were cytotoxic in both cell lines, while butanol phase and ellagic acid both with IC50>2048 µg/mL seemed to be safer. CONCLUSIONS: The results obtained indicate that the Excoecaria leaves can be conventionally used as antimicrobial, but it should be present that some cytotoxicity could appear. In addition, the three identified compounds were reported for the first time in the species. SUMMARY: Excoecaria lucida leaves (Euphorbiaceae) are used by the Cuban population due to their antimicrobial activity. This ethnopharmacological knowledge is confirmed by the integrated antibacterial and antifungal in vitro screening developed, using the bioassay-guided fractionation method.Abbreviations Used: MRC-5-SV2: Diploid human lung fibroblasts cells, RAW 264.7: Murine macrophages cells, IC50: Inhibitory Concentration 50%, ATCC: American Type Culture Collection, CCEBI: Culture Collection of Industrial Biotechnology Center, CECT: Spanish Culture Collection Type, CFU: Colony forming units, CC50: 50% cytotoxic concentration, CO2: Carbon dioxide, SI: Selectivity index, IR: Infrared spectroscopy, 1H NMR: Nuclear Magnetic Resonance of hydrogen, 13C NMR: Nuclear Magnetic Resonance of carbon, HMQC: Heteronuclear Multiple-Quantum Correlation, HMBC: Heteronuclear Multiple Bond Correlation, COSY: Correlation Spectroscopy, NOESY: Nuclear Overhauser Effect Spectroscopy, KBr: Potassium bromide, DMSO-D6: Deuterated dimethyl sulfoxide, LC.MS: Liquid Chromatography-Mass Spectrometry, [α]D: Optical rotation, EL1: ellagic acid, EL2: 3,3',4'-tri-O-methyl ellagic 4-O-ß-D-glucopyranoside acid, EL3: corilagin, Active (+), inactive (-).

Antioxidant and Hepatoprotective Activity of a New Tablets Formulation from Tamarindus indica L.

Hepatotoxic chemicals damage liver cells primarily by producing reactive oxygen species. The decoction of the leaves of Tamarindus indica L. is used for liver disorders. In this work we evaluated the hepatoprotective activity of a tablet formulation of this plant. Thirty-five Sprague Dawley rats were randomly divided into five groups (n = 7). First group (I) is control group, fed with standard diet. Groups II to V (hepatotoxic groups) were subjected to a subcutaneous injection of CCl4 (0.5 mL/kg). Group II was negative control, fed with standard diet; group III was subjected to administration of Silymarin 150 mg/kg and groups IV and V were treated with tablets in dose of 100 mg/kg and 200 mg/kg, respectively. Lipid peroxidation and the activity of superoxide dismutase, catalase, and reduced glutathione were evaluated. Serum levels of alanine aminotransferase, aspartate aminotransferase, gamma-glutamine transferase, alkaline phosphatase, and a lipid profile were evaluated too. The tablets inhibit lipid peroxidation. The redox balance (SOD-CAT-GSH) remains normal in the experimental groups treated with tablets. The liver function using dose of 200 mg/kg of tablets was better than the other experimental groups. These results justify, scientifically, the ethnobotanical use of the leaves of Tamarindus indica L.

Acute Toxicity and Dermal and Eye Irritation of the Aqueous and Hydroalcoholic Extracts of the Seeds of "Zapote" Pouteria mammosa (L.) Cronquist.

The common use of Pouteria mammosa (L.) Cronquist, "Mamey or Zapote," in food and ethnobotanic medicine shows its low or absent toxicity as fruit extracts prepared from seeds. However, it is essential to conduct security trials to scientifically support their use in drug therapy. This study evaluated the aqueous and hydroalcoholic extract (25%) Acute Oral Toxicity, obtained from the seeds of P. mammosa, in Sprague Dawley rats and dermal and eye irritability in New Zealand rabbits. The 404 and 405 acute dermal and eye irritation/corrosion guidelines were used, as well as the 423 Acute Oral Toxicity guideline, Acute Toxic Class Method of the Organization for Economic Cooperation and Development (OECD). The aqueous extract was located in the following category: not classified as toxic (CTA 5), while hydroalcoholic extract at 25% was classified as dangerous (CTA 4). Both extracts can be used without side reaction that irritates the skin which permitted classification as potentially not irritant. P. mammosa in the two extracts caused mild and reversible eye irritation, and it was classified as slightly irritating.

Chemical composition and insecticidal activity of Cymbopogon citratus essential oil from Cuba and Brazil against housefly.

Essential oil of Cymbopogon citratus collected from Brazil and Cuba was tested to a chemical characterization and then was tested on the post-embryonic development of Musca domestica. The chemical composition analysis by GC-MS of the oils from Brazil/Cuba allowed the identification of 13 and 12 major constituents respectively; nine of them common to both. In the both oils, the main components were the isomers geranial and neral, which together form the compound citral. This corresponds to a total of 97.92%/Brazil and 97.69%/Cuba of the compounds identified. The monoterpene myrcene, observed only in the sample of Cuba, presented a large relative abundance (6.52%). The essential oil of C. citratus (Brazil/Cuba) was dissolved in DMSO and tested at concentrations of 5, 10, 25, 50, 75 and 100% and citral was prepared by mixing 16.8 mg with 960 µL DMSO. Both essential oils and monoterpene citral were applied topically to newly-hatched larvae (1µL/larva). The results showed a lethal concentration (LC50) of 4.25 and 3.24% for the Brazilian and Cuban essential oils, respectively. Mortalities of larval and newly-hatched larvae to adult periods were dose-dependent for the two both oils as for monoterpene citral, reaching 90%. Both essential oils and citral caused morphological changes in adult specimens.