Extraction Of Anthocyanins From Arrabidaea Chica In Fixed Bed Using Co 2 And Co2/ethanol/water Mixtures As Solvents

Leaves of Arrabidaea chica (Humb. Bonpl.) Verlot are rich in anthocyanins and have been used as a medicinal plant in the Amazon region. In order to obtain different extracts from this plant, a sequential extraction in fixed bed was carried out at 40 °C and 300 bar, using supercritical carbon dioxide (scCO2) in a first step, and a mixture containing CO 2/ethanol/water at mass ratios of approximately 80/20/0, 80/14/6 and 80/10/10 in a second extraction step. The residue from the second step was extracted with water at 40 C and atmospheric pressure. Ethanolic, aqueous and hydroalcoholic (70:30, v:v) extracts were also obtained by conventional extraction methods at atmospheric pressure. All extracts were analyzed for global extraction yield, total phenolic content, total flavonoids, and carajurin content. High performance liquid chromatography (HPLC) was used both to quantify carajurin, which is the main anthocyanin component of A. chica, and to monitor qualitatively two other anthocyanin pigments found in that plant. The extraction yield in the first step was only 0.65% using pure scCO2, but this extraction was highly selective to extract carajurin from the three main anthocyanins. The accumulated global yield of the two steps ranged from 3% when the solvent ratio (80/20/0) was used in the second step to about 50% when 6 or 10% water was used, showing the highest yield when the extraction was done with water. The highest contents of total phenolic compounds (178 mg GAE/g extract) and total flavonoids (373 mg EC/g extract) were found in the process performed with the extraction mixture (80/20/0), and the highest carajurin content was obtained in the ethanolic extracts. © 2013 Elsevier B.V. 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Dissimilaridade genética em variedades de Artemisia annua L. embasada em caracteres agronômicos, fisiológicos e fitoquímicos

O presente estudo objetivou estimar a variabilidade genética existente entre caracteres agronômicos, fisiológicos e fitoquímicos em variedades de A. annua. O delineamento experimental foi inteiramente casualizado e os tratamentos foram as variedades Artemis, 2/39x5x3M, e 2/39x1V de A. annua, submetidas a avaliações agronômicas, fisiológicas e fitoquímicas. Para a realização das estimativas de distância genética foram geradas matrizes de dissimilaridade utilizando a distância Euclidiana e os métodos de agrupamento de Tocher e UPGMA. Além disso, avaliou-se a importância relativa dos caracteres para divergência genética pelo método de Singh. As análises foram realizadas pelo software Genes e os dendrogramas obtidos pelo NTSYS. A presença de variabilidade genética dentro das variedades permitiu a identificação de acessos dissimilares e com média elevada para as características estudadas. O número de ramificações, concentração intracelular de CO2, e o rendimento de óleo essencial foram os caracteres que mais contribuíram para a dissimilaridade genética de A. annua. Os acessos B24, C5 e C32 foram os mais promissores dentro das variedades e devem ser conservados para futuras hibridações, sendo que as hibridações mais promissoras na obtenção de populações segregantes desejadas são B24 x C5, B24 x C32 e C5 x C32

Genetic Dissimilarity In Varieties Of Artemisia Annua L. Based On Agronomic, Physiological And Phytochemical Characters [dissimilaridade Genética Em Variedades De Artemisia Annua L. Embasada Em Caracteres Agronômicos, Fisiológicos E Fitoquímicos]

This study aimed to estimate the genetic variability among agronomic, physiological, and phytochemical characters in varieties of A. annua. The experimental design was completely randomized and the treatments were the varieties Artemis, 2/39×5×3M and 2/39×1V of A. annua, subject to agronomic, physiological and phytochemical evaluations. To estimate the genetic distances, dissimilarity matrices were generated using the Euclidean distance and the Tocher and UPGMA grouping methods. Moreover, we evaluated the relative importance of the characters for genetic divergence through the method of Singh. The analyses were performed in the Genes software and the dendrograms were obtained from the NTSYS program. The presence of genetic variability within the varieties allowed the identification of dissimilar accessions with high average for all traits. The number of branches, intracellular concentration of CO2 and oil yield were the traits that contributed most to the genetic dissimilarity of A. annua. The accessions B24, C5 and C32 were the most promising within the populations and must be conserved for future crossings, and the most promising crosses to obtain the desired segregant populations were B24 × C5, B24 × C32 and C5 × C32.162 SUPPL. 1356363Arsenault, P.R., Recent Advances in Artemisinin Production Through Heterologous Expression (2008) Current Medicinal Chemestry, 15 (27), p. 2886Bagchi, G.D., Arteether: A potent plant growth inhibitor from Artemisia annua (1997) Phytochemistry., 45 (6), pp. 1131-1133Bertan, I., Dissimilaridade genética entre genótipos de trigo avaliados em cultivo hidropônico sob estresse por alumínio (2006) Bragantia, 65 (1), pp. 55-63Brisibe, E.A., Nutritional characterisation and antioxidant capacity of different tissues of Artemisia annuaL (2009) Food Chemistry, 115, pp. 1240-1246Celeghini, R.M.S., Desenvolvimento e validação de metodologia analítica por CLAE-IR para determinação de artemisinina em Artemisia annuaL (2009) Quimica Nova, 32 (4), pp. 875-878Charles, D.J., Simon, J.E., Germoplasm variation in artemisinin content of Artemisia annuausing na alternative method of artemisinin analysis from crude plant extracts (1990) Journal of Natural Products, 53 (1), pp. 157-160Cruz, C.D., (2001) Programa Genes:Aplicativo computacional em genética e estatística, p. 648. , Viçosa, Editora UFVDelabays, N., The genetics of artemisinin content in Artemisia annuaL. and the breeding of high yielding cultivars (2001) Current Medicinal Chemistry, 8 (15), pp. 1795-1801Efferth, T., Willmar Schwabe Award 2006: Antiplasmodial and Antitumor Activity of Artemisinin-From Bench to Beside (2007) Planta Medica, 73 (4), pp. 299-309Erdemoǧlu, N., Determination of Artemisinin in Selected Artemisia L. Species of Turkey by Reversed Phase HPLC (2007) Records of Natural Products, 1 (2-3), pp. 36-43Ferreira, J.F.S., Developmental studies of Artemisia annua: Flowering and artemisinin production under greenhouse and field conditions (1995) Planta Medica, 61 (2), pp. 167-170Ferreira, J.F.S., Janick, J., Distribution of artemisinin in Artemisia annua (1996) Progress in new crops, pp. 579-584. , In: JANICK, J. (Ed.). Arlington: ASHS PressFerreira, J.F.S., (2004) Artemisia annuaL.: The hope against malaria and câncer. Medicinal and aromatic plant: Production, business e applications, , http://www.ars.usda.gov/SP2UserFiles/person/34667/Ferreira-ArtemisiavsMalariaandCancer.pdf, Disponível em. Acesso em: 10 nov. 2010Ferreira, J.F.S., Luthria, D.L., Drying Affects Artemisinin, Dihydroartemisinic Acid, Artemisinic Acid, and the Antioxidant Capacity of Artemisia annuaL. Leaves (2010) Journal of Agricultural and Food Chemistry, 58 (3), pp. 1691-1708Geldre, E.V., State of the art of the production of the antimalarial compound artemisinin in plant (1997) Plant Molecular Biology, 33 (2), pp. 199-209Graham, I.A., The Genetic Map of Artemisia annua L. Identifies Loci Affecting Yield of the Antimalarial Drug Artemisinin (2010) Science, 327, pp. 328-331Holm, Y., Variation in the Essential Oil Composition of Artemisia annuaL. of Different Origin Cultivated in Finland (1997) Flavour and fragrance journal, 12, pp. 241-246Jain, D.C., Isolation of high artemisinin-yielding clones of Artemisia annua (1996) Phytochemistry, 43 (5), pp. 993-1001Lai, H., Singh, N.P., Oral artemisinin prevents and delays the development of 7,12-dimethylbenz[a]anthracene (DMBA)-induced breast cancer in the rat (2006) Cancer Letters, 231 (1), pp. 43-48Magalhães, P.M., (1996) Seleção, melhoramento e nutrição da Artemisia annua L., para cultivo em região intertropical, p. 132. , Tese (Doutorado). Universidade Estadual de Campinas, CampinasMagalhães, P.M., New hybrid lines of the antimalarial species Artemisia annuaL (1999) Acta Horticulturae, p. 502Magalhães, P.M., Yields of antimalarial Artemisia annuaL. species (2004) Acta Horticulturae, p. 629Marchese, J.A., Rehder, V.L.G., Influência da temperatura na produção de artemisinina em Artemisia annuaL (2001) Revista Brasileira de Plantas Medicinais, 4 (1), pp. 89-93Marchese, J.A., Flowering of Artemisia annua L. Plants Submitted to Different Photoperiod and Temperature Conditions (2002) Acta Horticulturae, p. 569Marchese, J.A., Carbon isotope composition and leaf anatomy as a tool to characterize the photosynthetic mechanism of Artemisia annuaL (2005) Brazilian Journal of Plant Physiology, 17 (1), pp. 187-190Marchese, J.A., (2006) Caracterização do mecanismo fotossintético e aspectos relacionados à floração de Artemisia annuaL, p. 68. , Tese (Doutorado). Departamento de Horticultura: Universidade Estadual Paulista, BotucatuOh, S., Synthesis and anti-cancer activity of covalent conjugates of artemisinin and a transferrin-receptor targeting peptide (2009) Cancer Letters, 274, pp. 33-39Sangwan, R.S., RAPD profile based genetic characterization of chemotypic variants of Artemisia annuaL (1999) Biochemistry and Molecular Biology International, 47 (6), pp. 935-944Singh, D., The relative importance of characters affecting genetic divergence (1981) The Indian Journal of Genetic and Plant Breeding, 41, pp. 237-24

In vitro and in vivo antitumor activity of crude extracts obtained from Brazilian Chromobacterium sp isolates

Natural products produced by microorganisms have been an important source of new substances and lead compounds for the pharmaceutical industry. Chromobacterium violaceum is a Gram-negative β-proteobacterium, abundant in water and soil in tropical and subtropical regions and it produces violacein, a pigment that has shown great pharmaceutical potential. Crude extracts of five Brazilian isolates of Chromobacterium sp (0.25, 2.5, 25, and 250 µg/mL) were evaluated in an in vitro antitumor activity assay with nine human tumor cells. Secondary metabolic profiles were analyzed by liquid chromatography and electrospray ionization mass spectrometry resulting in the identification of violacein in all extracts, whereas FK228 was detected only in EtCE 308 and EtCE 592 extracts. AcCE and EtCE 310 extracts showed selectivity for NCI/ADR-RES cells in the in vitro assay and were evaluated in vivo in the solid Ehrlich tumor model, resulting in 50.3 and 54.6% growth inhibition, respectively. The crude extracts of Chromobacterium sp isolates showed potential and selective antitumor activities for certain human tumor cells, making them a potential source of lead compounds. Furthermore, the results suggest that other compounds, in addition to violacein, deoxyviolacein and FK228, may be involved in the antitumor effect observed

Anthelmintic Activity Of Artemisia Annua L. Extracts In Vitro And The Effect Of An Aqueous Extract And Artemisinin In Sheep Naturally Infected With Gastrointestinal Nematodes

There is no effective natural alternative control for gastrointestinal nematodes (GIN) of small ruminants, with Haemonchus contortus being the most economically important GIN. Despite frequent reports of multidrug-resistant GIN, there is no new commercial anthelmintic to substitute failing ones. Although trematocidal activity of artemisinin analogs has been reported in sheep, neither artemisinin nor its plant source (Artemisia annua) has been evaluated for anthelmintic activity in ruminants. This study evaluated the anthelmintic activity of A. annua crude extracts in vitro and compared the most effective extract with artemisinin in sheep naturally infected with H. contortus. A. annua leaves extracted with water, aqueous 0.1 % sodium bicarbonate, dichloromethane, and ethanol were evaluated in vitro by the egg hatch test (EHT) and with the bicarbonate extract only for the larval development test (LDT) using H. contortus. The A. annua water, sodium bicarbonate (SBE), ethanol, and dichloromethane extracts tested in vitro contained 0.3, 0.6, 4.4, and 9.8 % of artemisinin, respectively. The sodium bicarbonate extract resulted in the lowest LC99 in the EHT (1.27 μg/mL) and in a LC99 of 23.8 μg/mL in the LDT. Following in vitro results, the SBE (2 g/kg body weight (BW)) and artemisinin (100 mg/kg BW) were evaluated as a single oral dose in naturally infected Santa Inês sheep. Speciation from stool cultures established that 84-91 % of GIN were H. contortus, 8.4-15.6 % were Trichostrongylus sp., and 0.3-0.7 % were Oesophagostomum sp. Packed-cell volume and eggs per gram (EPG) of feces were used to test treatment efficacy. The SBE tested in vivo contained no artemisinin, but had a high antioxidant capacity of 2,295 μmol of Trolox equivalents/g. Sheep dosed with artemisinin had maximum feces concentrations 24 h after treatment (126.5 μg/g artemisinin), which sharply decreased at 36 h. By day 15, only levamisole-treated sheep had a significant decrease of 97 % in EPG. Artemisinin-treated and SBE-treated sheep had nonsignificant EPG reductions of 28 and 19 %, respectively, while sheep in infected/untreated group had an average EPG increase of 95 %. Sheep treated with artemisinin and A. annua SBE maintained blood hematocrits throughout the experiment, while untreated/infected controls had a significant reduction in hematocrit. This is the first time oral dose of artemisinin and an aqueous extract of A. annua are evaluated as anthelmintic in sheep. Although oral dose of artemisinin and SBE, at single doses, were ineffective natural anthelmintics, artemisinin analogs with better bioavailability than artemisinin should be tested in vivo, through different routes and in multiple doses. The maintenance of hematocrit provided by artemisinin and A. annua extract and the high antioxidant capacity of the latter suggest that they could be combined with commercial anthelmintics to improve the well-being of infected animals and to evaluate potential synergism. © 2014 Springer-Verlag.113623452353Ademola, I.O., Eloff, J.N., In vitro anthelmintic activity of Combretum molle (R. Br. ex G. Don) (Combretaceae) against Haemonchus contortus ova and larvae (2010) Vet Parasitol, 169, pp. 198-203Bhakuni, R.S., Jain, D.C., Sharma, R.P., Kumar, S., Secondary metabolites of Artemisia annua and their biological activity (2001) Current Science, 80 (1), pp. 35-48Bin-Hafeez, B., Haque, R., Parvez, S., Pandey, S., Sayeed, I., Raisuddin, S., Immunomodulatory effects of fenugreek (Trigonella foenum graecum L.) extract in mice (2003) International Immunopharmacology, 3 (2), pp. 257-265. , DOI 10.1016/S1567-5769(02)00292-8Brisibe, E.A., Umoren, U.E., Brisibe, F., Magalhäes, P.M., Ferreira, J.F.S., Luthria, D., Wu, X., Prior, R.L., Nutritional characterisation and antioxidant capacity of different tissues of Artemisia annua L (2009) Food Chem, 115, pp. 1240-1246Celeghini, R.M.S., Sousa, I.M.O., Silva, A.P., Rodrigues, R.A.F., Foglio, M.A., Desenvolvimento e validação de metodologia analítica por CLAE-IR para determinação de Artemisinina em Artemisia annua L (2009) Quim Nova, 32, pp. 875-879Chagas, A.C.S., Vieira, L.S., Aragão, W.R., Navarro, A.M.C., Villela, L.C.V., Anthelmintic action of eprinomectin in lactating Anglo-Nubian goats in Brazil (2007) Parasitol Res, 100, pp. 391-394Chagas, A.C.S., Vieira, L.S., Freitas, A.R., Araújo, M.R.A., Araújo-Filho, J.A., Anthelmintic efficacy of neem (Azadirachta indica A. 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Lignocellulosic Biofuels and Grass Plants Used in Production of Pellets

Use of plant biomass in energy production is one ofthe preconditions to compensate growing shortage of fossilresources; therefore the research covered production of pelletsamples and testing of quality parameters thereof. The studyaimed at researching hemicellulose indicators in osier and poplarpellets, lignin content in perennial crops (depending on fertilisertypes, norms), parameters characterising pellet contentvariations (their evaluation and optimisation depending upon theproportions of pellet components) as well as characterisation oflignocellulose parameters

Healthcare access and quality index based on mortality from causes amenable to personal health care in 195 countries and territories, 1990-2015: A novel analysis from the global burden of disease study 2015

Background National levels of personal health-care access and quality can be approximated by measuring mortality rates from causes that should not be fatal in the presence of effective medical care (ie, amenable mortality). Previous analyses of mortality amenable to health care only focused on high-income countries and faced several methodological challenges. In the present analysis, we use the highly standardised cause of death and risk factor estimates generated through the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) to improve and expand the quantification of personal health-care access and quality for 195 countries and territories from 1990 to 2015. Methods We mapped the most widely used list of causes amenable to personal health care developed by Nolte and McKee to 32 GBD causes. We accounted for variations in cause of death certification and misclassifications through the extensive data standardisation processes and redistribution algorithms developed for GBD. To isolate the effects of personal health-care access and quality, we risk-standardised cause-specific mortality rates for each geography-year by removing the joint effects of local environmental and behavioural risks, and adding back the global levels of risk exposure as estimated for GBD 2015. We employed principal component analysis to create a single, interpretable summary measure-the Healthcare Quality and Access (HAQ) Index-on a scale of 0 to 100. The HAQ Index showed strong convergence validity as compared with other health-system indicators, including health expenditure per capita (r=0·88), an index of 11 universal health coverage interventions (r=0·83), and human resources for health per 1000 (r=0·77). We used free disposal hull analysis with bootstrapping to produce a frontier based on the relationship between the HAQ Index and the Socio-demographic Index (SDI), a measure of overall development consisting of income per capita, average years of education, and total fertility rates. This frontier allowed us to better quantify the maximum levels of personal health-care access and quality achieved across the development spectrum, and pinpoint geographies where gaps between observed and potential levels have narrowed or widened over time. Findings Between 1990 and 2015, nearly all countries and territories saw their HAQ Index values improve; nonetheless, the difference between the highest and lowest observed HAQ Index was larger in 2015 than in 1990, ranging from 28·6 to 94·6. Of 195 geographies, 167 had statistically significant increases in HAQ Index levels since 1990, with South Korea, Turkey, Peru, China, and the Maldives recording among the largest gains by 2015. Performance on the HAQ Index and individual causes showed distinct patterns by region and level of development, yet substantial heterogeneities emerged for several causes, including cancers in highest-SDI countries; chronic kidney disease, diabetes, diarrhoeal diseases, and lower respiratory infections among middle-SDI countries; and measles and tetanus among lowest-SDI countries. While the global HAQ Index average rose from 40·7 (95% uncertainty interval, 39·0-42·8) in 1990 to 53·7 (52·2-55·4) in 2015, far less progress occurred in narrowing the gap between observed HAQ Index values and maximum levels achieved; at the global level, the difference between the observed and frontier HAQ Index only decreased from 21·2 in 1990 to 20·1 in 2015. If every country and territory had achieved the highest observed HAQ Index by their corresponding level of SDI, the global average would have been 73·8 in 2015. Several countries, particularly in eastern and western sub-Saharan Africa, reached HAQ Index values similar to or beyond their development levels, whereas others, namely in southern sub-Saharan Africa, the Middle East, and south Asia, lagged behind what geographies of similar development attained between 1990 and 2015. Interpretation This novel extension of the GBD Study shows the untapped potential for personal health-care access and quality improvement across the development spectrum. Amid substantive advances in personal health care at the national level, heterogeneous patterns for individual causes in given countries or territories suggest that few places have consistently achieved optimal health-care access and quality across health-system functions and therapeutic areas. This is especially evident in middle-SDI countries, many of which have recently undergone or are currently experiencing epidemiological transitions. The HAQ Index, if paired with other measures of health-system characteristics such as intervention coverage, could provide a robust avenue for tracking progress on universal health coverage and identifying local priorities for strengthening personal health-care quality and access throughout the world. Copyright © The Author(s). Published by Elsevier Ltd