Ensayo de citotoxicidad de extractos de Pleurotusostreatus en diferentes líneas celulares para aplicaciones inmunonutricionales / Cytotoxicity assay of Pleurotus ostreatusextracts in different cell lines to immunonutritional applications / Cytotoxicity assay of Pleurotus ostreatusextracts in different cell lines to immunonutritional applications / Ensaio de citotoxicidade de extratos de Pleurotus ostreatus em diferentes linhagens celulares para aplicações imunonutricionais

Introducción: Los bioderivados propuestos como candidatos a ingredientes alimentarios suelen requerir ciertas evaluaciones para las aplicaciones inmunonutricionales Los hongos comestibles-medicinales son un surtidor de compuestos con estas potencialidades. Entre ellos, las setas Pleurotus ostreatus contienen metabolitos bioactivos, con importantes usos en la industria alimenticia y en la práctica terapéutica de la industria médico-farmacéutica. Los ensayos de citotoxicidad in vitro constituyen métodos valiosos para evaluarproductos de origen natural, como los extractos fúngicos. Objetivo: Evaluar la citotoxicidad de dos extractos obtenidos de la seta Pleurotus ostreatus en diferentes líneas celulares. Método: Se obtuvieron extractos hidrosolubles a partir del micelio y de los cuerpos fructíferos de Pleurotus ostreatus en laboratorios del Centro de Estudios de Biotecnología Industrial de la Universidad de Oriente. Se evaluó la citotoxicidad de los bioproductos por el ensayo de reducción del colorante resazurina sobre tres líneas celulares en el Laboratorio de Microbiología, Parasitología e Higiene (LMPH) de la Universidad de Amberes, Bélgica. Se utilizaron células no adherentes THP-1 (pre-monocitos de leucemia humana), células adherentes Caco-2 (epitelio de adenocarcinoma de colon humano) y células adherentes RAW 264.7 (macrófagos murinos). Resultados: Los extractos de Pleurotus ostreatus no resultaron citotóxicos para ninguna de las líneas celulares estudiadas humanas o murina, ya que no ocasionaron daños sobre la viabilidad de las célulasepiteliales del sistema gastrointestinal, nisobrelas células del sistema inmune empleadas. Conclusiones: Este resultado demuestra que ambos bioderivados fúngicos pueden ser aplicados con seguridad en estudios inmunonutricionales.(AU)

Introduction: Bioderivatives proposed as candidates for food ingredients usually require certain evaluations for immunonutritional applications. Edible-medicinal mushrooms are a source of compounds with these potentials. Among them, Pleurotus ostreatus mushrooms contain bioactive metabolites, with important uses in the food industry and in the therapeutic practice of the medical-pharmaceutical industry. In vitro cytotoxicity assays are valuable methods to evaluate products of natural origin, such as fungal extracts. Objective: To evaluate the cytotoxicity of two extracts obtained from the Pleurotus ostreatus mushroom in different cell lines. Method: Water-soluble extracts were obtained from the mycelium and fruiting bodies of Pleurotus ostreatus in laboratories of the Center for Industrial Biotechnology Studies of the Universidad de Oriente. The cytotoxicity of the bioproducts was evaluated by the resazurin dye reduction assay on three cell lines at the Laboratory of Microbiology, Parasitology and Hygiene (LMPH) of the University of Antwerp, Belgium. Non-adherent THP-1 cells (human leukemia pre-monocytes), Caco-2 adherent cells (human colon adenocarcinoma epithelium) and RAW 264.7 adherent cells (murine macrophages) were used. Results: Pleurotus ostreatus extracts were not cytotoxic for any of the human or murine cell lines studied, since they did not cause damage to the viability of the epithelial cells of the gastrointestinal system, nor to the immune system cells used. Conclusions: This result demonstrates that both fungal bioderivatives can be safely applied in immunonutritional studies.(AU)

Introdução: Bioderivados propostos como candidatos a ingredientes alimentícios geralmente requerem determinadas avaliações para aplicações imunonutricionais. Pleurotus ostreatus contêm metabólitos bioativos, com importantes utilizações na indústria alimentícia e na prática terapêutica da indústria médico-farmacêutica. Ensaios de citotoxicidade in vitro são métodos valiosos para avaliar produtos de origem natural, como extratos de fungos. Objetivo: Avaliar a citotoxicidade de dois extratos obtidos do cogumelo Pleurotus ostreatus em diferentes linhagens celulares. Método: Extratos hidrossolúveis foram obtidos do micélio e dos corpos frutíferos de Pleurotus ostreatus nos laboratórios do Centro de Estudos de Biotecnologia Industrial da Universidade de Oriente. A citotoxicidade dos bioprodutos foi avaliada pelo ensaio de redução do corante resazurina em três linhagens celulares no Laboratório de Microbiologia, Parasitologia e Higiene (LMPH) da Universidade de Antuérpia, Bélgica. Foram utilizadas células THP-1 não aderentes (pré-monócitos de leucemia humana), células aderentes Caco-2 (epitélio de adenocarcinoma do cólon humano) e células aderentes RAW 264.7 (macrófagos murinos). Resultados: Os extratos de Pleurotus ostreatus não foram citotóxicos para nenhuma das linhagens celulares humanas ou murinas estudadas, pois não causaram danos à viabilidade das células epiteliais do sistema gastrointestinal, nem às células do sistema imunológico utilizadas. Conclusões: Este resultado demonstra que ambos os bioderivados fúngicos podem ser aplicados com segurança em estudos imunonutricionais.(AU)

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.

Evaluación in vivo de la pirogenicidad de bioproductos fúngicos con potencial prebiótico / In vivo pyrogenicity evaluation of fungal bioproducts with prebiotic potential / Avaliação in vivo da pirogenicidade de bioprodutos fúngicos com potencial prebiótico

RESUMEN Introducción: Los productos naturales con actividad farmacológica requieren de evaluaciones preclínicas que justifiquen su empleo sobre una base científica. El ensayo de pirógenos es una prueba dentro de la Farmacología de Seguridad que se realiza para determinar la presencia de endotoxinas y constituye un método valioso, para demostrar la seguridad de bioderivados con potencial prebiótico en el campo de la inmunonutrición. Objetivo: Evaluar la pirogenicidad de bioproductos fúngicos de Pleurotus ostreatus (extractos acuosos del micelio y cuerpos fructíferos) y un biopreparado de levadura Kluyveromyces marxianus, empleando el ensayo de pirógenos en conejos Nueva Zelanda. Método: Se ensayaron concentraciones de 1,0 y 10,0 mg/mL de cada muestra por vía endovenosa en dosis de 0,5 y 5,0 mg/kg de peso. El diseño experimental cumplió las buenas prácticas de laboratorio según lo establecido por el International Council for Laboratory Animals Science y se realizó de acuerdo a los procedimientos normalizados de trabajo del Centro de Toxicología y Biomedicina, Santiago de Cuba. Resultados: Los extractos de Pleurotus ostreatus y el biopreparado de levadura (0,5 mg/kg) no mostraron signos de pirogenicidad. En los resultados del biopreparado (5,0 mg/kg), los valores de temperatura caen en un rango de incertidumbre, según la Farmacopea de Estados Unidos (USP) y se sugirió repetir el estudio. Conclusiones: Los extractos de Pleurotus ostreatus y el biopreparado de Kluyveromyces marxianus (0,5 mg/kg) no indujeron un aumento de temperatura significativo en los animales, lo cual sugiere que en estos bioproductos no existen niveles de endotoxinas que puedan provocar pirogenicidad.

ABSTRACT Introduction: Natural products with pharmacological activity require preclinical evaluations to justify their uses scientifically. The pyrogen assay is a safety pharmacology test performed to determine the presence of endotoxins and it is a valuable method to demonstrate the bio-derivative products safety and their prebiotic potential in the field of immunonutrition. Objective: To evaluate the pyrogenicity of fungal bioproducts from Pleurotus ostreatus (aqueous extracts from mycelium and fruiting bodies) and a biopreparation from Kluyveromyces marxianus yeast, using a pyrogen assay in New Zealand rabbits. Method: Concentrations of 1.0 and 10.0 mg/mL of each sample were tested intravenously at doses of 0.5 and 5.0 mg/kg body weight. The experimental design complied with good laboratory practices as established by the International Council for Laboratory Animal Science and was carried out according to the standard work procedures of the Centro de Toxicología y Biomedicina, Santiago de Cuba. Results: Pleurotus ostreatus extracts and the yeast biopreparation (0.5 mg/kg) showed no signs of pyrogenicity. In the biopreparation results (5.0 mg/kg), temperature values fall in the uncertainty range according to the United States Pharmacopoeia (USP), and therefore it was suggested to repeat the study. Conclusions: Pleurotus ostreatus extracts and Kluyveromyces marxianus biopreparation (0.5 mg/kg) did not induce a significant temperature increase in the animals, thereby suggesting that there are no endotoxin levels in such bioproducts that could cause pyrogenicity.

RESUMO Introdução: Produtos naturais com atividade farmacológica requerem avaliações pré-clínicas que justifiquem seu uso em bases científicas. O ensaio de pirogênio é um teste dentro da Farmacologia de Segurança que é realizado para determinar a presença de endotoxinas e é um método valioso para demonstrar a segurança de bioderivados com potencial prebiótico no campo da imunonutrição. Objetivo: Avaliar a pirogenicidade de bioprodutos fúngicos de Pleurotus ostreatus (extratos aquosos do micélio e corpos de frutificação) e de uma biopreparação da levedura Kluyveromyces marxianus, utilizando o ensaio pirogênico de coelho da Nova Zelândia. Método: Concentrações de 1,0 e 10,0 mg/mL de cada amostra foram testadas por via intravenosa nas doses de 0,5 e 5,0 mg/kg de peso. O desenho experimental obedeceu às boas práticas laboratoriais estabelecidas pelo Conselho Internacional para a Ciência dos Animais de Laboratório e foi realizado de acordo com os procedimentos de trabalho padrão do Centro de Toxicologia e Biomedicina, Santiago de Cuba. Resultados: Os extratos de Pleurotus ostreatus e a biopreparação de leveduras (0,5 mg/kg) não apresentaram sinais de pirogenicidade. Nos resultados da biopreparação (5,0 mg/kg), os valores de temperatura estão dentro de uma faixa de incerteza, segundo a Farmacopeia dos Estados Unidos (USP) e foi sugerido repetir o estudo. Conclusões: Os extratos de Pleurotus ostreatus e a biopreparação de Kluyveromyces marxianus (0,5 mg/kg) não induziram um aumento significativo da temperatura nos animais, o que sugere que não há níveis de endotoxinas nesses bioprodutos que possam causar pirogenicidade.

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.

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.

Differential Induction Pattern Towards Classically Activated Macrophages in Response to an Immunomodulatory Extract from Pleurotus ostreatus Mycelium.

Pleurotus ostreatus mushroom preparations have been investigated because of their ability to modulate the immune function. However, there is still no consensus regarding the activation and polarizing effect on macrophages by Pleurotus-derived bioproducts. This study examined the immune-activating effect of a mycelium-derived P. ostreatus aqueous extract (HW-Pm) on macrophage functions, by means of the determination of nitric oxide (NO) production, the mRNA expression of inducible nitric oxide synthase (iNOS), Arginase-1 and FIZZ and the cytokine levels. The phagocytic activity and the activation of NF-κB in U937 reporter cells were also investigated. No cytotoxicity was observed in macrophages treated with HW-Pm (IC50 > 1024 µg/mL) by the resazurin test. HW-Pm induced high levels of NO production and iNOS expression in macrophages. In contrast, HW-Pm did not induce Arginase-1 and FIZZ mRNA expressions. The mushroom extract increased TNF-α and IL-6 production and the phagocytic function in murine macrophages. It also stimulated the activation of the NF-κB promoter. The P. ostreatus mycelium extract has a potential application as a natural immune-enhancing agent, by targeting macrophage activation towards the classically activated subset and stimulating macrophage-mediated innate immune responses.

COMPOSICIÓN MICOQUÍMICA Y ACTIVIDAD ANTIOXIDANTE DE LA SETA Pleurotos ostreatus EN DIFERENTES ESTADOS DE CRECIMIENTO / Mycochemical composition and antioxidant activity of Pleurotus ostreatus mushroom in different growth stages

RESUMEN Pleurotus sp. es un género de basidiomicetos ampliamente distribuido a nivel mundial, con especies que revisten importancia, no solo desde el punto de vista comercial por su valor nutricional, sino también por sus propiedades medicinales. Sin embargo, aún son insuficientes los estudios micoquímicos y se desconoce su relación con las propiedades antioxidantes de bioproductos obtenidos de estas setas durante diferentes estados de crecimiento. En el presente trabajo, se determinó el contenido de proteínas, carbohidratos, azúcares reductores, fenoles totales y flavonoides, así como la actividad antioxidante in vitro (ensayos de captación de radicales DPPH y estimación del poder reductor) de extractos acuosos de Pleurotus ostreatus en dos estados de crecimiento (primordios y cuerpos fructíferos maduros) durante la fermentación sólida. El contenido de proteínas, azúcares reductores y flavonoides fue superior en el extracto acuoso de cuerpos fructíferos maduros, mientras que los carbohidratos y fenoles totales fueron mayores en el extracto de primordios. Los valores de EC50 en los ensayos de DPPH y los de poder reductor fueron de 1,72 mg/mL y 3,27 mg/mL para el extracto de primordios y de 0,44 mg/mL y 0,48 mg/mL para el de cuerpos fructíferos maduros, respectivamente. Los resultados sugieren que las propiedades antioxidantes de extractos acuosos de Pleurotus ostreatus (primordios y cuerpos fructíferos maduros) reflejan las variaciones en la concentración de moléculas bioactivas, teniendo en cuenta las particularidades fisiológicas de las fases de crecimiento. Ello podría resultar de utilidad para el diseño de protocolos de obtención de bioproductos de Pleurotus ostreatus con aplicaciones potenciales como antioxidantes en las industrias alimentaria y farmacéutica.

ABSTRACT Pleurotus sp. is a genus of Basidiomycetes widely distributed worldwide, with species that are important not only from the commercial point of view for its nutritional value, but also for their medicinal properties. However, mycochemical studies and the relationship with the antioxidant properties of bioproducts obtained from these mushrooms during different growth stages are still insufficient. In this work, the content of proteins, carbohydrates, reducing sugars, total phenols and flavonoids was determined, as well as the in vitro antioxidant activity (tests of scavenging DPPH and reducing power estimation) of aqueous extracts of Pleurotus ostreatus in different growth stages (primordia and mature fruiting bodies) during the solid fermentation. The content of proteins, reducing sugars and flavonoids was higher in the aqueous extract ofmature fruiting bodies, while carbohydrates and total phenols were increased in the extract ofprimordia. The EC50 values in the DPPH and reducing power assays were 1.72 mg/mL and 3.27 mg/mL for primordia extract and 0.44 mg/mL and 0.48 mg/mL in the case of mature fruiting bodies extract, respectively. The results suggest that the antioxidant properties of Pleurotus ostreatus aqueous extracts (primordia and mature fruiting bodies), reflected the variations in the concentration of bioactive molecules, taking into account the physiological characteristics of the growth phases. This could be useful in designing protocols for obtaining bioproducts from Pleurotus ostreatus with potential applications as antioxidant in food and pharmaceutical industries.

Micromonospora fluminis sp. nov., isolated from mountain river sediment.

During a bioprospection of bacteria with antimicrobial activity, the actinomycete strain A38T was isolated from a sediment sample of the Carpintero river located in the Gran Piedra Mountains, Santiago de Cuba province (Cuba). This strain was identified as a member of the genus Micromonospora by means of a polyphasic taxonomy study. Strain A38T was an aerobic Gram-positive filamentous bacterium that produced single spores in a well-developed vegetative mycelium. An aerial mycelium was absent. The cell wall contained meso-diaminopimelic acid and the whole-cell sugars were glucose, mannose, ribose and xylose. The major cellular fatty acids were isoC15:0, 10 methyl C17:0, anteiso-C17:0 and iso-C17:0. The predominant menaquinones were MK-10(H4) and MK-10(H6). Phylogenetic analysis of 16S rRNA gene sequences revealed that this strain was closely related to Micromonospora tulbaghiae DSM 45142T (99.5 %), Micromonospora citrea DSM 43903T (99.4 %), Micromonospora marina DSM 45555T (99.4 %), Micromonospora maritima DSM 45782T (99.3 %), Micromonospora sediminicola DSM 45794T (99.3 %), Micromonospora aurantiaca DSM 43813T (99.2 %) and Micromonospora chaiyaphumensis DSM 45246T (99.2 %). The results of OrthoANIu analysis showed the highest similarity to Micromonospora chalcea DSM 43026T (96.4 %). However, the 16S rRNA and gyrB gene sequence-based phylogeny and phenotypic characteristics provided support to distinguish strain A38T as a novel species. On the basis of the results presented here, we propose to classify strain A38T (=LMG 30467T=CECT 30034T) as the type strain of the novel species Micromonospora fluminis sp. nov.

Nocardiopsis dassonvillei subsp. crassaminis subsp. nov., isolated from freshwater sediment, and reappraisal of Nocardiopsis alborubida Grund and Kroppenstedt 1990 emend. Nouioui et al. 2018.

An actinomycete, strain D1T, was isolated from a freshwater sediment sample collected from the San Pablo river in the La Risueña community, Santiago de Cuba province, Cuba. The strain was identified as a member of the genus Nocardiopsis by means of a polyphasic taxonomic study. It produced a light yellow non-fragmented substrate mycelium, a white well-developed aerial mycelium and straight to flexuous hyphae. No specific spore chains were observed. Strain D1T contained meso-diaminopimelic acid, no diagnostic sugars, and MK-10(H2), MK-10(H4), MK-10 and MK-10(H6) as predominant menaquinones, but not phosphatidylcholine as diagnostic polar lipid of the genus Nocardiopsis. The predominant fatty acids were iso-C16 : 0, 10-methyl-C18 : 0 and anteiso-C17 : 0. Strain D1T showed the highest degree of 16S rRNA gene sequence similarity to Nocardiopsis synnematoformans DSM 44143T (99.8 %), Nocardiopsis dassonvillei subsp. albirubida NBRC 13392T (99.8 %) and Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111T (99.6 %). A genomic OrthoANIu value between D1T and N. dassonvillei subsp. dassonvillei DSM 43111T of 97.63 % and a dDDH value of 78.9 % indicated that strain D1T should be classified in N. dassonvillei. However, phenotypic characteristics distinguished strain D1T from its nearest neighbour taxon. On basis of these results we propose to classify strain D1T (=LMG 30468T=CECT 30033T) as a representative of a novel subspecies of the genus Nocardiopsis, for which the name Nocardiopsis dassonvillei subsp. crassaminis subsp. nov. is proposed. In addition, the genomic distance between N. dassonvillei subsp. albirubida NBRC 13392T and N. dassonvillei subsp. dassonvillei DSM 43111T as determined through OrthoANIu (93.64 %) and dDDH (53.40 %), along with considerable phenotypic and chemotaxonomic differences reported in earlier studies, indicated that the classification of this taxon as Nocardiopsis alborubida Grund and Kroppenstedt 1990 is to be preferred over its classification as N. dassonvillei subsp. albirubida Evtushenko et al. 2000.

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.

Plantas y hongos comestibles en la modulación del sistema inmune / Eatable plants and mushroom in the immune system modulation

Introducción: Se presenta una revisión que considera resultados de investigaciones que han centrado su atención en la búsqueda de compuestos obtenidos de plantas medicinales y hongos comestibles, capaces de estimular la respuesta inmunitaria. Objetivos: Recopilar y actualizar la información científica publicada acerca de las plantas medicinales y hongos comestibles activos sobre el sistema inmune. Métodos: Se realizó una búsqueda bibliográfica de artículos científicos relacionados con la temática, en las bases de datos disponibles (PubMed, EBSCO, SciELO, LILACS, CUMED, MEDNAT) así como varios libros y sitios web. Resultados: En los estudios realizados se refiere como las principales plantas con actividad inmunomoduladora: Allium sativum (Liliaceae), Aloe vera (Aloaceae), Uncaria tomentosa (Rubiaceae), Morinda citrifolia (Rubiaceae), Echinacea purpurea (Asteraceae) y Mangifera indica (Anacardiaceae). Diversos hongos comestibles activos sobre el sistema inmune han sido examinados: Ganoderma lucidum (Ganodermataceae), Grifola frondosa (Polyporaceae), Lentinus edodes (Agaricaceae), Pleurotus spp. (Pleurotaceae) y Schizophylum commune (Schizophyllaceae), entre otros. Asimismo se destacan las posibilidades que estas fuentes naturales podrían ofrecer en la manipulación de la respuesta inmune, al aumentar la calidad de vida de pacientes con inmunodeficiencias y cáncer. Conclusiones: El estudio de productos naturales derivados de plantas y hongos comestibles-medicinales ofrece amplias perspectivas para el desarrollo de nuevos agentes inmunomoduladores, eficaces y seguros. Se debe continuar trabajando en la identificación de los principios activos y en la dilucidación del mecanismo mediante el que actúan

Introduction: Present review asses the results of researches basing its attention in the search of compounds obtained from medicinal plants and mushrooms, able to stimulate the immune response. Objectives: To collect and to update the scientific information published on the medicinal plants and active mushrooms on immune system. Methods: A bibliographic search of scientific papers related to this subject matter in the available databases (PubMed, EBSCO, SciELO, LILACS, CUMED, MEDNAT) as well as some books and websites. Results: In studies conducted it is mentioned how the main plants with immunomodulation activity: Allium sativum (Liliaceae), Aloe vera (Aloaceae), Uncaria tomentosa (Rubiaceae), Morinda citrifolia (Ribiaceae), Echinacea purpurea (Asteraceae) and Mangifera indica (Anacardiacear). Different active mushrooms on the immune system have been examined: Ganoderma lucidum (Ganodermataceae), Frifola frondosa (Polyporaceae), Lentinus edodes (Agaricaceae), Pleurotus spp. (Pleurotaceae) and Schizophylum commune (Schizophyllaceae) among others. Likewise, authors emphasized the possibilities that these natural sources could offer in management of immune response increasing the quality of life of patients presenting with immunodeficiencies and cancer. Conclusions: The study of natural products derived of medicinal plants and mushrooms offers many perspectives for the development of new effective and safe immunomodulation agents. It is necessary to continue working in the identification of the active principles and in the elucidation of the mechanism by means of they act

Aplicación de la técnica de aclaramiento del carbón coloidal en sangre periférica en la evaluación de la actividad inmunomoduladora de dos productos naturales / Applications of the colloidal carbón clearence technique in peripheral blood in the evaluation of the immunomodulator activity of two natural products

En el presente trabajo se evaluó el efecto de dos productos naturales en la actividad in vivo de los macrófagos esplénicos y las células de Kupffer, en diferentes sistemas experimentales, por medio de la prueba de aclaramiento de carbón coloidal en sangre periférica. Los resultados evidenciaron el efecto estimulador de un hidrolizado proteico de la microalga Chlorella vulgaris 87/1 sobre el sistema monocito-macrófago, administrado como suplemento a ratones malnutridos, y de extractos obtenidos de setas comestibles Pleurotus spp. en tres biomodelos de inmunodeficiencias secundarias (inducida por ciclofosfamida, por exposición a radiaciones ionizantes y por malnutrición proteico-energética). En los animales tratados con los productos se observó un menor tiempo de vida media de las partículas de carbón coloidal en sangre, lo que es una evidencia de la actividad inmunopotenciadora de ambos biopreparados y de la aplicabilidad de la técnica en cuestión.

In this paper, the effect of two natural products in the activity in vivo of the splenic macrophages and the Kupffer's cells was evaluated in different experimental systems by means of the colloidal carbon clearance test in peripheral blood. The results evidenced the stimulating effect of a protein hydrolizate of the microalga Chlorella vulgaris 87/1 on the monocyte-macrophagic system, administered as a supplement to malnourished mice, and of extracts obtained from edible mushrooms Pleurotus spp. in three biomodels of secondary immunodeficiencies (induced by cyclophosphamide, by exposure to ionizing radiations and by protein-energy malnutrition). In the animals treated with the products, it was observed a shorter time of mean life of the particles of colloidal carbon in blood, which shows the immunopotentiating activity of both biopreparations and of the applicability of the technique.