Producción de biomasa y caracterización química de Lentinula edodes cultivado en biorreactor bajo diferentes condiciones

La ingeniería bioquímica juega un papel importante en el desarrollo tecnológico de los procesos de obtención ya sea de cuerpos fructíferos de setas con potencial medicinal para el consumo directo, o en la fabricación de nutracéuticos y/o suplementos dietarios. Esta investigación implementó una metodología en biorreactor para el cultivo del hongo macromiceto Lentinula edodes, conocido comúnmente como Shiitake, con el objetivo de evaluar el efecto que tienen la velocidad de agitación y el flujo de aire en la producción de biomasa y de metabolitos de mediana y baja polaridad. Se determinó que el rendimiento y la productividad más altos de producción de biomasa se logra a unas condiciones de 80 rpm y 0,50 vvm obteniendo 0,0260 g/g y 0,00433 g/L*h respectivamente. Igualmente, en cuanto a la composición de esta biomasa, se concluyó que se genera un máximo contenido de proteína (33,4% del micelio liofilizado) a unas condiciones de 140 rpm y 0,75 vvm, los esteroles alcanzaron un porcentaje de 89,32% del extracto en DCM a 80 rpm y 0,5 vvm. Los resultados del análisis por GC-MS confirman la amplia variedad de compuestos que se pueden obtener a partir de un cultivo en biorreactor del hongo Lentinula edodes.

Biochemical engineering plays an important role in the technological development of the processes for obtaining either mushroom fruiting bodies with medicinal potential for direct consumption, or in the manufacture of nutraceuticals and/or dietary supplements. This research implemented a methodology in a bioreactor for the cultivation of the Lentinula edodes macromycete fungus, commonly known as Shiitake, with the aim of evaluating the effect of agitation speed and air flow on the production of biomass and metabolites of median and low polarity. Thus, it was found that the highest yield and productivity of biomass production is achieved at conditions of 80 rpm and 0,50 vvm, obtaining 0,0260 g/g and 0,00433 g/L*h respectively. Likewise, in the study of the content of metabolites it was concluded that for the protein 33,4% of the lyophilized mycelium was reached at conditions of 140 rpm and 0,75 vvm, the sterols reached a percentage of 89,32% of the extract. in DCM at 80 rpm and 0,5 vvm. The results of the GC-MS analysis confirm the wide variety of compounds that can be obtained from a bioreactor culture of the Lentinula edodes fungus.

The biochemical content and antioxidant capacities of endemic Tanacetum densum (Lab.) Schultz Bip. subsp. laxum, and Tanacetum densum (Lab.) Schultz Bip. subsp. amani Heywood growing in Turkey / O conteúdo bioquímico e as capacidades antioxidantes de Tanacetum densum endêmico (lab.) Schultz Bip. subsp. laxum e Tanacetum densum (lab.) Schultz Bip. subsp. amani Heywood que crescem na Turquia

Abstract Medicinal plants have a significant role in preventing and curing several diseases, and Tanacetum L. is one of these plants. The aim of the present study is to determine the fatty acid, lipid-soluble vitamin, sterol, phenolic content, and antioxidant capacity of Tanacetum densum subsp. laxum and Tanacetum densum subsp. amani, to compare the effect of altitude on the biochemical content and to compare systematically by using fatty acids and phenolics. This study showed that palmitic acid (C16:0) and stearic acid (C18:0) are major sources of saturated fatty acid and oleic acid (C18:1 n9), and linoleic acid (18:2 n6c) and a-linolenic acid (C18:3 n3) are the principal unsaturated fatty acids in the two endemic Tanacetum densum taxa. Also, this study found that the unsaturated fatty acid content (60.11±1.61%) of Tanacetum densum subsp. laxum was higher than the unsaturated fatty acid content (44.13±1.28%) of Tanacetum densum subsp. amani. And also, the ω6/ω3 ratio of Tanacetum densum subsp. laxum (1.74) and Tanacetum densum subsp. amani (1.60) was found to be similar. However, this study determined that the lipid soluble vitamin and sterol content of two endemic Tanacetum taxa are low except for stigmasterol. Present study showed that catechin is principal phenolic in the Tanacetum densum taxa. This study also found that Tanacetum densum subsp. laxum and Tanacetum densum subsp. amani had the highest levels of catechin, vanillic acid, and caffeic acid content though the phenolic amounts, particularly catechin and quercetin, were dissimilar in the T. densum taxa. This study suggested that ecological conditions such as altitude may affect the biochemical content of two endemic Tanacetum densum taxa. Furthermore, the current study determined that two endemic Tanacetum L. taxa had potent radical scavenging capacities and found a correlation between total phenolics and antioxidant activity.

Resumo As plantas medicinais têm um papel significativo na prevenção e cura de várias doenças, e Tanacetum L. é uma dessas plantas. O objetivo do presente estudo é determinar o conteúdo de ácido graxo, vitamina lipossolúvel, esterol, estrutura fenólica e capacidade antioxidante de Tanacetum densum subsp. laxum e Tanacetum densum subsp. amani, comparar o efeito da altitude sobre o conteúdo bioquímico e realizar uma comparação sistemática usando ácidos graxos e fenólicos. Esta pesquisa mostrou que o ácido palmítico (C16:0) e o ácido esteárico (C18:0) são as principais fontes de ácido graxo saturado e que o ácido oleico (C18:1 n9), o ácido linoleico (18:2 n6c) e o ácido a-linolênico (C18: 3 n3) são os principais ácidos graxos insaturados nos dois táxons endêmicos de Tanacetum densum. Além disso, este estudo descobriu que o conteúdo de ácidos graxos insaturados (60,11±1,61%) de Tanacetum densum subsp. laxum foi superior ao conteúdo de ácidos graxos insaturados (44,13±1,28%) de Tanacetum densum subsp. amani, e também que a razão ω6/ω3 de Tanacetum densum subsp. laxum (1,74) e Tanacetum densum subsp. amani (1,60) foi semelhante. No entanto, este trabalho determinou que o conteúdo de vitamina lipossolúvel e esterol de dois táxons endêmicos de Tanacetum é baixo, exceto o estigmasterol, além de descobrir que Tanacetum densum subsp. laxum e Tanacetum densum subsp. amani apresentaram os mais altos níveis de conteúdo de catequina, ácido vanílico e ácido cafeico, embora as quantidades fenólicas, especialmente catequina e quercetina, sejam diferentes nos táxons de T. densum. Este estudo sugere que condições ecológicas, como a altitude, podem afetar o conteúdo bioquímico de dois táxons endêmicos de Tanacetum densum. Ainda, esta pesquisa determinou que dois táxons de Tanacetum L. endêmicos possuíam potentes capacidades de sequestro de radicais e que houve correlação entre fenólicos totais e atividade antioxidante.

The SREBP (Sterol Regulatory Element-Binding Protein) pathway: a regulatory bridge between carotenogenesis and sterol biosynthesis in the carotenogenic yeast Xanthophyllomyces dendrorhous

Xanthophyllomyces dendrorhous is a basidiomycete yeast that naturally produces the red-orange carotenoid astaxanthin, which has remarkable antioxidant properties. The biosynthesis of carotenoids and sterols share some common elements that have been studied in X. dendrorhous. For example, their synthesis requires metabolites derived from the mevalonate pathway and in both specific pathways, cytochrome P450 enzymes are involved that share a single cytochrome P450 reductase, CrtR, which is essential for astaxanthin biosynthesis, but is replaceable for ergosterol biosynthesis. Research on the regulation of carotenoid biosynthesis is still limited in X. dendrorhous; however, it is known that the Sterol Regulatory Element-Binding Protein (SREBP) pathway, which is a conserved regulatory pathway involved in the control of lipid metabolism, also regulates carotenoid production in X. dendrorhous. This review addresses the similarities and differences that have been observed between mammal and fungal SREBP pathways and what it is known about this pathway regarding the regulation of the production of carotenoids and sterols in X. dendrorhous.

Analysis of Chemical Composition in Sancao Baogan Decoction Based on UPLC-ESI-HRMSn / 中国实验方剂学杂志

Objective::To analysis the chemical constituents in Sancao Baogan decoction by ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-ESI-HRMSn). Method::The separation was performed on an ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) by a gradient elution of methanol (A)-0.1% formic acid solution (B) (0-2 min, 5%A; 2-20 min, 5%-12%A; 20-35 min, 12%-40%A; 35-38 min, 40%A; 38-48 min, 40%-80%A; 48-50 min, 80%A). The flow rate was 0.3 mL·min-1 and the column temperature was set at 30 ℃, the injection volume was 10 μL. Electrospray ionization was applied and the data were collected via positive and negative ion modes. By using Xcalibur 3.0 software, the chemical constituents were analyzed based on the relative retention time, excimer ion peak and fragment ion peak of the compounds, as well as comparison with human metabolome database (HMDB), reference substances and literature data. Result::A total of 40 chemical components were identified from Sancao Baogan decoction, including 16 phenolic acids, 19 flavonoids, 2 anthraquinones, 1 triterpenoid, 1 sterol, and 1 monoterpenoid. Six compounds (dansensu, α-pinene, epigallocatechin, 2, 5-dihydroxybenzoic acid, naringenin and emodin) were reported for the first time from Sancao Baogan decoction. Conclusion::The established UPLC-ESI-HRMSn can quickly, accurately and comprehensively analyze the chemical constituents of Sancao Baogan decoction, which lays a foundation for the basic research of pharmacodynamic substances and quality control of this formula.

Phytosterols Content in Vegetable Oils of Brazil: Coconut, Safflower, Linseed and Evening Primrose

Abstract In the last years phytosterols, natural components of plants, have received more attention due to association of their consumption with reducing risk of cardiovascular diseases and cancer. There are several scientific studies about phytosterols in vegetable oils, but they are scarce in unconventional oils. The objective of this research was evaluating the content of phytosterols (β-sitosterol, stigmasterol and campesterol) in vegetable oils sold in São Paulo city, in Brazil. The analysis included cold alkaline saponification, derivatization with hexamethyldisilazane and trimethylchlorosilane reagents, and quantification by gas chromatography using flame ionization detection and internal standardization. The quality control parameters indicated that the method was suitable for analysis. Total sterols were between 272.3 mg kg-1 (coconut oil) to 6169.7 mg kg-1 (evening primrose oil). β-sitosterol was the component found in higher concentrations and evening primrose oil was the most representative in quantity of phytosterols.

Study on chemical constituents from whole herbs of Sargassum fusiforme / 中草药

Objective: To study the chemical constituents of the whole herbs of Sargassum fusiforme. Methods: The chemical constituents from S. fusiforme were separated and purified by silica gel, MCI, ODS, Sephadex LH-20 gel column chromatographies, and preparative HPLC. Their structures were identified by analysis of NMR, MS, ORD, etc, and comparing physicochemical properties of compounds with references. Results: A total of 19 compounds were obtained from the 50% ethanol-H2O extract of S. fusiforme, and their structures were determined as lupenone (1), bauerenone (2), β-amyrenone (3), α-amyrenone (4), loliolide (5), isololiolide (6), (3S,5R,6S,7E)-5,6-epoxy-3-hydroxy-7-megastigmen-9-one (7), (R)-dehydrovomifoliol (8), 24-ethylcholesta-4,24 (28)-dien-3-one (9), fucosterol (10), 29-hydroperoxystigmasta-5,24 (28)-dien-3β-ol (11), 24-hydroperoxy-24-vinylcholesterol (12), saringosterol (13), (24R)-5,28-stigmastadiene-3β,24-diol-7-one (14), dihydroquercetin (15), aurantiamide acetate (16), dibutyl phthalate (17), glycerol monopalmitate (18), and methyl hexadecanoate (19). Conclusion: The compounds isolated from S. fusiforme were mainly identified as triterpenoids, sesquiterpenoids, highly oxidized sterols, alkaloids and so on. Compounds 1-4, 7-8 and 14-19 are isolated from S. fusiforme for the first time.

A new lignan from Amauroderma rude / 中草药

Objective: Study on the chemical constituents in the ethyl acetate phase extracted from the fruiting bodies of Amauroderma rude. Methods: Silica gel column chromatography, Sephadex LH-20 gel and liquid chromatography were used to separate and purify the chemical constituents in fruiting bodies of Amauroderma rude. The compounds were identified according to their physical and chemical properties and spectroscopic data. Results: Three compounds including one new lignan and two known sterols were isolated from the ethyl acetate phase of this fungus. Their structures were identified as 3,4-dihydroxyphenacyl acetyl ketone (1), (22E, 24R)-3β,5α-dihydrox-yerogosta-7,22-diene-6-one (2), and (22E,24R)-ergosta-6,9,22-trien-3β,5α,8α-triol (3). Conclusion: Compound 1 was a new lignan and named amaurolignan B.

Protective effects of Taraxasterol on oxidatively injured cardiomyocytes / 重庆医学

Objective To observe the protective effect and mechanism of taraxasterol on cardiomyocytes in oxidative injury model caused by ischemia-reperfusion (I/R).Methods Mouse cardiomyocytes (CSC cells) were used as the study objects and divided into the normal control group,I/R group,taraxasterol treating I/R group (5,10,30 μmol/L) and positive control group.The cell viability was measured by MTT.The expressions of Caspase-3 and Bcl-2 were detected by RT-PCR.The expressions of superoxide dismutase (SOD) and malondialdehyde (MAD) were detected by biochemical methods.The expression of ERK1/2 was detected by western blot.Results MTT assay showed that 30 μmol/L taraxasterol increased the cell viability of CSC cells injured by I/R (P＜0.05).The RT-PCR results showed that the expression of Caspase-3 mRNA was decreased with 10,30 μmol/L taraxasterol treatment,the difference was statistically significant when compared with I/R group (P＜0.05).The expression of Bcl-2 mRNA was increased with 30 μmol/L taraxasterol treatment,the difference was statistically significant when compared with the I/R group (P＜0.05).The biochemical method detection showed that 30 μmol/L taraxasterol induced SOD expression was increased and MAD expression was decreased,the difference was statistically significant when compared with the I/R group (P＜0.05).Western blot detection showed that 30 μmol/L taraxasterol treatment increased the ratio of p-ERK1/2 to t-ERK1/2 in injured CSC cells (P＜0.05).Conclusion Taraxasterol might inhibit ischemia-reperfusion caused cardiomyocyte oxidative injury by up-regulation of ERK1/2 expression.

Euphorbia umbellata bark extracts – an in vitro cytotoxic study

ABSTRACT Euphorbia umbellata (Pax) Bruyns, Euphorbiaceae, is commonly used in folk medicine of southern Brazil to treat several kinds of cancer. The latex (part of the plant used for this purpose) is mixed with water and taken as treatment; but this matrix contains toxic potential related to the presence of some phorbol type diterpenes. So the aim of this study was to evaluate the cytotoxicity of the crude extract of the bark of E. umbellata and its fractions (Hex, CHCl3, EtOAc and MeOH) using in vitro assay (applying Jurkat cells line). A preliminary cytotoxic study (MTT reduction, trypan blue exclusion and DNA quantification assays) was executed to identify the most active material. The CHCl3 fraction displayed the highest activity and was selected for further investigation of any cytotoxic mechanism and evaluation of chemical composition; flow cytometry, Acridine orange and Hoechst 33342 staining experiments and Gas chromatography–mass spectrometry analysis were applied to achieve these results. This fraction demonstrated the best cytotoxic results against Jurkat cells line with IC50 of 29.00 ± 1.49, 10.06 ± 1.48 and 4.83 ± 2.25 µg/ml for 24, 48 and 72 h of experiment, respectively (trypan blue exclusion). The mechanism responsible for this action can be associated with the promotion of cell cycle arrest and apoptosis. The two main classes of compounds present in the CHCl3 fraction are steroids and triterpenes. Further, phytochemical studies with this fraction need to be evaluated, to try isolating these substances and establishing a more detailed cytotoxic study against Jurkat cells.

Research progress on chemical constituents and pharmacological of Cirsium japonicum / 中草药

Major chemical constituents of Cirsium japonicum are flavonoids, sterols, lignans, lonicol, glycosides, and volatile oils, which has hemostatic, hemostatic, antitumor and other pharmacological effects. By searching the researches in SciFinder, Chinese knowledge network, and PubMed from 1977 to 2017, the previous studies on C. japonicum were summarized and analyzed from chemical composition and pharmacological effect, which provided reference for the development and basic research of pharmacodynamic substances.

De novo assembly and transcriptome characterization of Penthorum chinense / 中草药

Objective In order to obtain the reference sequences and relative expression of transcription genes and study the genetic base of active ingredients in Penthorum chinense, which were useful for researching functional gene to P. chinense. Methods In this study, by performing Illumina Hiseq 2000 and de novo assembly, the transcriptome of whole plant was sequenced, the data were filtered and assembled, and the unigene was compared and annotated. Meanwhile, the genes related to the synthesis of metabolic pathway of active ingredients in P. chinense were analyzed. Results Totally, 40 005 442 valid short sequences were obtained, and 42 306 unigenes were spliced by de novo. Also, a total of 518 open reading frames (ORF) were obtained by ORF analysis, and 75 ORF of them had transcription factor domains. In addition, by performing KEGG pathway analysis, 33, 32, 59, and 68 unigenes were found to be involved in the pathway of flavonoid biosynthesis, steroid biosynthesis, terpenoid backbone biosynthesis, and 2-oxocarboxylic acid metabolism, respectively. Conclusion The datasets provided in this study will contribute significantly to genetic improvement and study on the genes related to biosynthesis pathway of pharmaceutical active substances from P. chinense.

Chemical Constituents from Mycelia and Spores of Fungus Cordyceps cicadae / 中草药·英文版

Objective To isolate and identify the chemical constituents from mycelia and spores of the fungus Cordyceps cicadae, respectively. Methods The chemical constituents were isolated and purified by repeated silica gel, Sephadex LH-20, and reversed phase HPLC. The structures of these compounds were elucidated on the basis of extensive spectroscopic analysis, including 1D and 2D NMR. Results Nine known sterols such as ergosterol (1), ergosterol peroxide (2), 9,11-dehydroergosterol peroxide (3), 3β,5α,9α-trihydroxy-(22E,24R)-ergosta-7,22-dien-6-one (4), 3β,5α,9α,14α-tetrahydroxy-(22E, 24R)-ergosta-7,22-dien-6-one (5), 5α,6α-epoxy-(22E,24R)-ergosta-8(14),22-diene-3β,7α-diol (6), 3β,5α,6β-(22E,24R)-ergosta-7,22-dien-3,5,6-triol (7), 3β,5α,6α-6-methoxyergosta-(22E,24R)-7,22-diene-3,5-diol (8), 4-hydroxy-17R-methylincisterol (9), together with a resorcinol derivative, 5-n-nonadecylresorcinol (10), a cyclodesipeptide, beauvericin (11), and a nucleoside, N6-(2-hydroxyethyl)adenosine (12) were successively isolated from the cultivated C. cicadae mycelia and spores. Conclusion Compounds 3–10 are reported for the first time from the title sample, beauvericin exhibits significant cytotoxicity against human leukemia cell line and human lung cancer cell line.

Combined bioactive approach over atherosclerosis risk biomarkers / Abordagem combinada de compostos bioativos sobre biomarcadores de risco para aterosclerose

Atherosclerosis, one major cause of morbidity and mortality worldwide, is a complex and multifactorial disease that involves three mainly conditions: chronic inflammation, dyslipidemia and oxidative stress. Although statins are the first-line therapy for LDL cholesterol (LDL-C) lowering, the efficacy of cardiovascular events prevention is limited to 30-40%. This residual risk brought attention to the need of new therapies and clinical targets beyond LDL-C, such as inflammation and oxidative stress. Importantly, suboptimal treatment and/or statin discontinuation due to adverse effects have also been a very challenging clinical problem. Complementary diet therapy can be an effective and safe approach to support pharmacological treatment, especially when drugs alone are insufficient to attenuate risk factors and/or the recommended dose is not well tolerated. The aim of this study was to evaluate the effects of three bioactive components, namely omega-3 fatty acids, plant sterols and polyphenols, on markers of dyslipidemia, inflammation and oxidative stress in patients treated with statins. A randomized, crossover clinical study was carried out, with the participation of 53 subjects. At each intervention period, study participants received a packaged for the functional or control treatment. Functional treatment consisted of fish oil (1.7 g of EPA+DHA/day), chocolate containing plant sterols (2.2 g/day) and green tea (two tea sachets/day). Control treatment consisted of soy oil softgels, regular chocolate and anise tea. After 6 weeks of intervention, functional treatment reduced plasma LDL-C (-13.7% ± 3.7, p=0.002) and C-reactive protein (-35.5% ± 5.9, p=0.027). Plasma triacylglycerol (-15.68% ± 5.94, p=0.02) and MDA (-40.98% ± 6.74, p=0.04) were reduced in subgroups of patients (n=23) with baseline values above the median (93 mg/dL and 2.23 umol/L, respectively). Analysis of lathosterol and campesterol in plasma suggested that intensity of LDL-C reduction was influenced by cholesterol absorption rate rather than its endogenous synthesis. After multivariate analysis, patients identified as "good responders" to supplementation (n=10) were recruited for a pilot protocol of statin dose reduction with complementary diet therapy. Responders received the functional treatment for 12 weeks: standard statin therapy was kept during the first 6 weeks and reduced by 50% from weeks 6 to 12. No difference was observed for plasma lipids and inflammation biomarkers, cholesterol efflux capacity or HDL particle number after statin dose reduction when compared to standard therapy. Although limited by the small sample size, our study demonstrates the potential for a new therapeutic approach combining lower statin dose and specific dietary compounds. This may be particularly helpful for the many patients with, and at risk for, CVD who cannot tolerate high-dose statin therapy

A aterosclerose, uma importante causa mundial de morbidade e mortalidade, é uma doença complexa e multifatorial que envolve três principais condições: inflamação crônica, dislipidemia e estresse oxidativo. Embora as estatinas sejam fármacos de primeira linha para redução de LDL colesterol (LDL-C), sua eficácia na prevenção de eventos cardiovasculares é limitadada a 30-40%. Este risco cardiovascular residual evidencia a necessidade de novas terapias e marcadores clínicos que vão além do LDL-C, como inflamação e estresse oxidativo. Não obstante, tratamento subótimo e/ou interrupção do uso de estatinas devido à ocorrencia de efeitos adversos também é um grave obstáculo na clínica médica. Neste contexto, a terapia dietética complementar representa uma abordagem efetiva e segura para o suporte do tratamento farmacológico, especialmente quando as drogas são insuficientes para atenuar fatores de risco e/ou quando a dose recomendada não é bem tolerada. O objetivo do presente estudo foi avaliar o efeito de três compostos bioativos - ácidos graxos ômega 3, fitosteróis e polifenóis - sobre marcadores de inflamação, lipemia e estresse oxidativo em indivíduos tradados com estatinas. Foi realizado um estudo clínico randomizado, de delineamento crossover, com a participação de 53 voluntários. A cada período de intervenção, os participantes receberam um tratamento funcional ou controle. O tratamento funcional foi composto por cápsulas de óleo de peixe (1.7 g/dia de EPA+DHA), chocolate contendo fitosteróis (2.2 g/dia) e chá verde (dois sachês/dia). O tratamento controle foi composto por cápsulas de óleo de soja, chocolate sem adição de fitosteróis e chá de anis. Após 6 semanas de intervenção, o tratamento funcional reduziu a concentração plasmática de LDL-C (-13.7% ± 3.7, p=0.002) e proteína C-reativa (-35.5% ± 5.9, p=0.027). Triglicerídeos (- 15.68% ± 5.94, p=0.02) e malondialdeído (-40.98% ± 6.74, p=0.04) foram reduzidas apenas em subgrupos de indivíduos que apresentavam valores basais acima da mediana (93 mg/dL e 2.23 umol/L, respectivamente). A análise de latosterol e campesterol no plasma sugeriu que a intensidade da redução de LDL-C não foi influenciada pela síntese endógena de colesterol, mas sim pela taxa de absorção. Após análise multivariada dos resultados, pacientes identificados como "good responders" à suplementação (n=10) foram recrutados para um estudo piloto de redução da dosagem da estatina, aliado à terapia dietética complementar. Estes pacientes receberam o tratamento funcional por 12 semanas: durante as 6 primeiras semanas mantevese a dosagem de estatina, que em seguida foi reduzida em 50% das semanas 6 a 12. Não foram observadas diferenças para os marcadores plasmáticos de lipídeos, inflamação, capacidade de efluxo de colesterol ou número de partículas de HDL após a redução da dose de estatina, quando comparada à terapia convencional. Embora limitado pelo reduzido número de pacientes, o estudo demonstra o potencial para uma nova abordagem terapêutica, combinando reduzida dose de estatina com específicos compostos bioativos. Esta pode ser uma importante alternativa para muitos pacientes em risco cardiovascular e que são intolerantes à terapia com altas doses de estatina.

Composición lipídica y evaluación de las actividades antioxidante y leishmanicida del basidiomiceto Ganoderma sp / Lipid composition and evaluation of the antioxidant and leishmanicidal activities of the basidiomycete Ganoderma sp

Introducción: los hongos del género Ganoderma han sido utilizados para el cuidado de la salud en la medicina tradicional asiática por más de 2000 años. Desde 1980 los estudios químicos han reportado un sin número de metabolitos secundarios con propiedades bioactivas. Objetivo: identificar compuestos lipídicos en el extracto etanólico del hongo Ganoderma sp., además de evaluar sus actividades antioxidante y leishmanicida. Métodos: la extracción de las fracciones lipídicas presentes en el cuerpo fructífero de Ganoderma sp. Se realizó por Cromatografía en Columna. La elucidación estructural se determinó por Espectrometría de Masas y Resonancia Magnética Nuclear. La actividad antioxidante del extracto etanólico fue evaluada con las metodologías del radical 2,2-difenil-1-picrilhidrazil (DPPH) y el radical catiónico 2,2'-azinobis (3-etilbenzotiazolina-6-ácido sulfónico) (ABTS); la actividad leishmanicida por citometría de flujo y la actividad citotóxica usando el ensayo colorimétrico de bromuro de 3-(4,5-dimetil-tiazol-2-il)-2,5-difenil tetrazolio (MTT) sobre la línea celular U937. Resultados: diecinueve esteres metílicos y ergosterol fueron identificados por espectrometría de masas en el extracto etanólico. Un compuesto triterpenoidal se elucidó usando Espectroscopia de Resonancia Magnética Nuclear. Los valores de concentración media inhibitoria (IC 50) de la actividad antioxidante del extracto etanólico para las metodologías de los radicales DPPH y ABTS fueron de 85,63 µg/mL y 62,82 µg/mL, respectivamente. Los valores de las actividades citotóxica y leishmanicida fueron > 200,0 µg/mL y 21,5 µg/mL ± 4,4 respectivamente. Conclusiones: las estructuras de los derivados de ácidos grasos elucidados corresponden a compuestos con diferentes grados de insaturación. En este estudio se realizó el reporte de la Ganoderona A, como compuesto triterpenoidal. La elevada actividad antioxidante en relación a otros trabajos sugiere que este organismo es una fuente importante de metabolitos secundarios con propiedades captadoras de radicales libres, aunque los valores de actividad leishmanicida no fueron significativos se recomienda continuar con el estudio de otras particiones del extracto etanólico(AU)

Introduction: Fungi from the genus Ganoderma have been used in Asian traditional medicine for more than 2 000 years. Since the year 1980 chemical studies have reported a large number of secondary metabolites with bioactive properties. Objective: Identify lipid compounds in ethanolic extract from the fungus Ganoderma sp. and evaluate their antioxidant and leishmanicidal activities. Methods: Extraction of lipid fractions from the fruiting body of Ganoderma sp. was conducted by column chromatography. Structural features were determined by mass spectrometry and nuclear magnetic resonance. Antioxidant activity of the ethanolic extract was evaluated with the methodologies for radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and cationic radical 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS); leishmanicidal activity by flow cytometry, and cytotoxic activity with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay (MTT) on cell line U937. Results: Nineteen methyl esters and ergosterol were identified by mass spectrometry in the ethanolic extract. A triterpenoid compound was identified by nuclear magnetic resonance spectroscopy. Mean inhibitory concentration values (IC50) for antioxidant activity of the ethanolic extract using the methodologies for radicals DPPH and ABTS were 85.63 µg/ml and 62.82 µg/ml, respectively. Values for cytotoxic and leishmanicidal activities were > 200.0 µg/ml and 21.5 µg/ml ± 4.4, respectively. Conclusions: The structure of the fatty acid derivatives identified corresponds to compounds with varying degrees of unsaturation. The study included the report of Ganoderma A as a triterpenoid compound. Antioxidant activity was found to be higher than in previous studies, suggesting that this organism is an important source of secondary metabolites with free radical scavenging properties. Although leishmanicidal activity values were not found to be significant, it is recommended to study other partitions of the ethanolic extract(AU)

Phytosterols isolated from endophytic fungus Colletotrichum gloeosporioides (Melanconiaceae) / Fotoesteróis isolados do fungo endofítico Colletotrichum gloeosporioides (Melanconiaceae)

Endophytic fungi are fungi that colonize internal tissues of plants. There are few studies of compounds isolated from endophytic fungi of Amazon plants. Thus, the aim this study was the isolation and structural identification of sitosterol (1), stigmasterol (2), sitostenone (3), squalene (4), ergosterol (5) and ergosterol peroxide (6) from fungus Colletotrichum gloeosporioidesisolated as endophytic from Virola michelli, a typical Amazon plant, used in folk medicine against skin infection. Compounds were isolated by chromatography column on silica and identified by 1H and 13C NMR and MS. The presence of phytosterols in fungi is rare and this is the first report of the isolation of the phytosterols sitosterol, stigmasterol and sitostenone from the genus Colletotrichum.

Fungos endofíticos são fungos que colonizam os tecidos internos das plantas. Existem poucos estudos de compostos isolados de fungos endofíticos de plantas da Amazônia. Assim, o objetivo deste estudo foi o isolamento e identificação estrutural de sitosterol (1), estigmasterol (2), sitostenona (3), esqualeno (4), ergosterol (5) e peroxido de ergosterol (6) do fungo Colletotrichum gloeosporioidesisolado como endofítico de Virola michelli, uma planta típica da Amazônia, usada na medicina popular no combate a infecções de pele. Os compostos foram isolados por cromatografia em coluna de sílica e identificados por RMN 1H e 13C e EM. A presença de fitoesteróis em fungos é rara e este é o primeiro relato do isolamento dos fitoesteróides sitosterol, estigmasterol e sitostenona do gêreno Colletotrichum.

Study on the Chemical Constituents of Sedum lineare / 中国药房

OBJECTIVE:To study the chemical constituents of Sedum lineare. METHODS:Silica gel column chromatogra-phy,TLC and crystallization were adopted to isolate the chemical constituents of S. lineare. And chemical structures were analyzed and identified based on physicochemical properties and spectral data of compounds. RESULTS:A total of 6 sterols and 2 triterpe-noids were isolated from petroleum ether fraction of S. lineare,which were identified as stigmaster-5-ene-3β-ol-7-one(1),stigmas-ter-5-ene-3β,7α-diol(2),daucosterol(3),daucosterol palmitate(4),β-sitosterol(5),stigmaster-7-ene-3β-ol(6),δ-amyrin(7),andδ-amyrone(8)respectively. CONCLUSIONS:Compounds 3,4 and 7 are isolated from S. lineare for the first time,and compound 4 is isolated from genus S.lineare for the first time. The study has laid certain foundation for the quality evaluation of S. lineare.

Studies on constituents from rhizome of Arundo donax / 中草药

Objective: To study chemical components of rhizome of Arundo donax, a folk medicine. Methods: Using different methods such as chromatography and recrystallization purification to get chemical components, and the structures were identified by physical and chemical properties and spectral data. Results: Twenty-three compounds were isolated and identified as following: hexadecanoic acid (1), n-docosane (2), myristic acid glycerides (3), 5,6-epoxy-22,24-ergosta-8(14),22-diene-3,7-diol (4), 5,6-epoxy-22,24-ergosta-8(9),22-diene-3,7-diol (5), 5,8-epidioxy-22,24-ergosta-6,22-dien-3-ol (6), stigmast-4-ene-3,6-dione (7), 6,9-epoxy-ergosta-7,22-dien-3-ol (8), stigmast-22-en-3,6, 9-triol (9), 3,4,5-trimethoxyphenol (10), 2,6-dimethoxy-1,4-quinone (11), sinapaldehyde (12), hydroxycinnamic acid (13), β-sitostenone (14), α-asarone (15), 4-dodecylbenzaldehyde (16), β-sitosterol (17), α-spinasterol (18), p-hydroxybenzaldehyde (19), ursolic acid (20), N-acetyltryptamine (21), daucosterol (22), and (-)-syringaresinol (23). Conclusion: Compouds 1-16 and 18-23 are isolated from the plant in genus Arundo L. for the first time.

Cloning and protein expression of sterol-C-24-methyl transferase 2 in Tripterygium wilfordii / 药学学报

24-Alkyl sterols are the major players in the control of membrane component and plant growth. In this paper, we cloned an important rate-limiting enzyme:sterol-C-24-methyl transferase (SMT) in the sterol biosynthetic pathway according to the transcriptome data of Tripterygium wilfordii. suspension cells, whose full-length cDNA was 1 631 bp with an open reading frame of 1 080 bp, encoding a protein of 359 amino acids. It was estimated that theoretical isoelectric point (pI) was 6.43 and the molecular mass was 40.0 kDa. Bioinformatics analysis attributed the SMT gene to SMT2 family. The expression vector was constructed as the pMAL-c2x-TwSMT2 plasmid and the recombinant protein was expressed in E. coil BL21 (DE3) competent cells. After methyl jasmonate treatment, the relative expression level of TwSMT2 has improved significantly in 24 h. SDS-PAGE electrophoresis and Western Blot showed that protein of TwSMT2 in BL21 (DE3) strain was expressed after induction by IPTG. In this study, TwSMT2 was cloned for the first time and the recombinant protein was expressed, which lay the foundation for elucidation of the sterol biosynthetic pathway of Tripterygium wilfordii in the future.

Anti-inflammatory and analgesic activities with gastroprotective effect of semi-purified fractions and isolation of pure compounds from Mediterranean gorgonian Eunicella singularis

OBJECTIVE@#To explore anti-inflammatory activities of organic extract and its semi-purified fractions (ethanol, acetone, methanol/dichloromethane) from the Mediterranean gorgonian Eunicella singularis.@*METHODS@#The anti-inflammatory and analgesic activities were evaluated, using the carrageenan-induced rat paw edema model and the acetic acid writhing test in mice. The gastroprotective activity was determined using HCl/EtOH induced gastric ulcers in rats. The purification and structure elucidation of compound(s) from the more effective fraction were determined by chromatographic and spectroscopic methods and in comparison with data reported in the literature.@*RESULTS@#The fraction F-EtOH showed an important anti-inflammatory activity associated with significant analgesic and gastroprotective properties. The purification and structure elucidation of compound(s) from this fraction lead to the identification of one diterpenoid and four sterols.@*CONCLUSIONS@#These results suggested that components from the active fraction can be used to treat various anti-inflammatory diseases.

Anti-inflammatory and analgesic activities with gastroprotective effect of semi-purified fractions and isolation of pure compounds from Mediterranean gorgonian Eunicella singularis

Objective: To explore anti-inflammatory activities of organic extract and its semi-purified fractions (ethanol, acetone, methanol/dichloromethane) from the Mediterranean gorgonian Eunicella singularis. Methods: The anti-inflammatory and analgesic activities were evaluated, using the carrageenan-induced rat paw edema model and the acetic acid writhing test in mice. The gastroprotective activity was determined using HCl/EtOH induced gastric ulcers in rats. The purification and structure elucidation of compound(s) from the more effective fraction were determined by chromatographic and spectroscopic methods and in comparison with data reported in the literature. Results: The fraction F-EtOH showed an important anti-inflammatory activity associated with significant analgesic and gastroprotective properties. The purification and structure elucidation of compound(s) from this fraction lead to the identification of one diterpenoid and four sterols. Conclusions: These results suggested that components from the active fraction can be used to treat various anti-inflammatory diseases.