Lecythis Pisonis Research Articles

Assessment of subcritical propane, supercritical CO2 and Soxhlet extraction of oil from sapucaia (Lecythis pisonis) nuts

The extraction of sapucaia (Lecythis pisonis) nut oil (SNO) using subcritical propane (SPE) and supercritical CO2 (with ethanol as co-solvent; scCO2) as solvent was investigated and compared with the conventional (Soxhlet) extraction. Extraction with scCO2 was performed at 333K and 20MPa while the SPE extractions were carried out in different conditions to investigate the effects of temperature (293–333K) and pressure (2–10MPa) on the oil yield and the chemical compositions of the products. Results show that SPE allowed a fast extraction with a higher yield (46.22%) obtained at 333K and 10MPa, representing 93% efficiency compared to Soxhlet. Only temperature had significant (p<0.05) effect on the extraction yield. SPE yielded the oils with highest values of polyunsaturated fatty acids (∼36%). Stability to oxidation ranges from 6.53 to 11.17h. The major triacylglycerols present in SNO are OOO, SOO, POO, PLO, and POS.

Cuticular n-alkane in leaves of seven Neotropical species of the family Lecythidaceae: a contribution to chemotaxonomy

ABSTRACT Biosynthesized from very long-chain fatty acid wax precursors, n-alkanes make a valuable contribution to the taxonomy of plants. The alkane components of foliar epicuticles of seven Neotropical species of Lecythidaceae were investigated: Bertholletia excelsa, Cariniana legalis, Couroupita guianensis, Eschweilera alvimii, Eschweilera ovata, Gustavia augusta and Lecythis pisonis. Specimens were collected in the metropolitan area of Recife, Pernambuco, and their n-alkane fractions were analyzed by gas chromatography. The chemical relationships among the species were then evaluated using cophenetic correlation and UPGMA. Among the seven species, a total of 15 n-alkanes, with 21-35 carbon atoms, were identified and formed a consistent group of B. excelsa, C. guianensis, E. ovata, G. augusta, and L. pisonis with n-C31. The greatest similarities were found between B. excelsa and L. pisonis, and between C. guianensis and G. augusta. Nevertheless, a phenetic analysis based on a larger number of species is needed to better understand the chemotaxonomic value of epicuticular n-alkanes within the Lecythidaceae.

Neuroprotective effect of Sapucaia nuts (Lecythis pisonis)on rats fed with high-fat diet.

Lecythis pisonis Cambess is commonly known as "castanha de sapucaia" in Brazil. Chemical composition studies revealed that this nut is an excellent source of anti-oxidant minerals and of essential lipids. The aim of the present study is to assess the anti-oxidant and anti-inflammatory effect of Lecythis pisonis Cambess on the brain tissue of Wistar rats. The animals were divided in four experimental groups (n = 6), total of forty-eight rats. Treatments included the standard diet (AIN-93G) and high-fat food, supplemented with Sapucaianut from 14 to 28 days. The gene expression markers TNF-α, NFkB, ZnSOD and HSP-72 were defined through reverse transcriptase polymerase chain reaction (rtPCR). The anti-oxidant effect was assessed through the thiobarbituric acid-reactive substances (TBARS) and the measurement of the activity performed by superoxide dismutase enzymes. Accordingly, the gene expression of the inflammatory markers NFkB (p65) and TNF-αwas lower in rats fed on diets supplemented with "sapucaia", and they presented significant difference in the Tukey test (p < 0.05). The heat-shock HSP-72 protein and the ZnSOD enzyme raised the gene expression and showed significant statistical difference (p < 0.05) in both groups fed on Sapucaia nut-based diet. Thus, the nutritional properties of the Sapucaia nuts perform important neuroprotective activities because they modulated the anti-oxidant activity and the brain tissue inflammatory process in the assessed animals.

English

Lecythis pisonis Cambess is commonly known as “sapucaia” nut. Previous studies show that it is rich in unsaturated fatty acids and in antioxidant minerals. The aim of the present study was to assess the antioxidant and anti-inflamatory effects of this nut after its introduction into a control (AIN-93G) or high-fat diet in Wistar rats. The animals were divided into four groups: a control diet, the same control diet supplemented with sapucaia nuts, a high-fat diet or the high-fat diet supplemented with sapucaia nuts and were fed with these diets for 14 or 28 days. The gene expression of the markers tumor necrosis factor (TNF)-α NFκB (p65) zinc superoxide dismutase (ZnSOD) and heat shock protein 72 (HSP72) was determined by the chain reaction to the quantitative reverse transcription-polymerase chain reaction (q-PCR). The antioxidant activity was also measured as thiobarbituric acid reactive substances (TBARS) through the activity of the SOD enzyme. The groups treated with “sapucaia” nuts presented reduced lipid peroxidation values and increased ZnSOD and HSP72 gene expression, as well as decreased TNF-α and NFκB (p65) gene expression levels. The significant results showed that “sapucaia” could serve as a potential source of antioxidants and as a protector agent for the examined animals. Key words: Sapucaia nuts, inflammation, oxidative stress, gene expression.

Lecythis marcgraaviana (Lecythidaceae), an overlooked species from eastern Brazil

Lecythis marcgraaviana (Lecythidaceae), a species previously placed in synonymy of L. pisonis, is resurrected and described in detail for the first time. A neotype, discussion of the history of this species, taxonomic notes, images, and an identification key to the species of the Lecythis pisonis clade from eastern Brazil are provided. This tree or shrub occurs in coastal Atlantic Forest between the states of Espirito Santo and Rio Grande do Norte. Morphologically, L. marcgraaviana is most similar to L. lanceolata and L. pisonis.

Isolation, characterization and mechanism of action of an antimicrobial peptide from Lecythis pisonis seeds with inhibitory activity against Candida albicans.

Antimicrobial peptides (AMPs) are produced by a range of organisms as a first line of defense against invaders or competitors. Owing to their broad antimicrobial activity, AMPs have attracted attention as a potential source of chemotherapeutic drugs. The increasing prevalence of infections caused by Candida species as opportunistic pathogens in immunocompromised patients requires new drugs. Lecythis pisonis is a Lecythydaceae tree that grows in Brazil. The AMPs produced by this tree have not been described previously. We describe the isolation of 12 fractions enriched in peptides from L. pisonis seeds. Of the 12 fractions, at 10 μg/ml, the F4 fraction had the strongest growth inhibitory effect (53.7%) in Candida albicans, in addition to a loss of viability of 94.9%. The F4 fraction was separated into seven sub-fractions by reversed-phase chromatography. The F4.7' fraction had the strongest activity at 10 μg/ml, inhibiting C. albicans growth by 38.5% and a 69.3% loss of viability. The peptide in F4.7' was sequenced and was found to be similar to plant defensins. For this reason, the peptide was named L. pisonis defensin 1 (Lp-Def1). The mechanism of action that is responsible for C. albicans inhibition by Lp-Def1 includes a slight increase of reactive oxygen species induction and a significant loss of mitochondrial function. The results described here support the future development of plant defensins, specifically Lp-Def1, as new therapeutic substances against fungi, especially C. albicans.

Toward a phylogenetic-based Generic Classification of Neotropical Lecythidaceae—I. Status of Bertholletia, Corythophora, Eschweilera and Lecythis

Lecythidaceae subfam. Lecythidoideae is limited to the Neotropics and is the only naturally occurring subfamily of Lecythidaceae in the New World. A subset of genera with zygomorphic flowers—Bertholletia, Corythophora, Eschweilera and Lecythis—comprises a group of about 125 species called the Bertholletia clade. A previous study based on plastid ndhF and trnL-F genes supported the monophyly of Corythophora but suggested that Eschweilera and Lecythis are not monophyletic. Using this study as a baseline, we sampled more taxa and sequenced more loci to address the taxonomic problems of the ambiguous genera and to determine relationships within the Bertholletia clade. Our results support the monophyly of the Bertholletia clade as previously circumscribed. In addition, Corythophora is monophyletic, and the two accessions of Bertholletia excelsa come out together on the tree. Results of the simultaneous analysis do not support the monophyly of Lecythis or Eschweilera. Lecythis consists of four main groups (the Lecythis pisonis, L. poiteaui, L. chartacea, and L. corrugata clades), the last of which is nested within Eschweilera, and Eschweilera consists of three clades (the Eschweilera integrifolia, E. tetrapetala, and Eschweilera parvifolia clades). We compare our results with the generic classification presented in the latest monograph of neotropical Lecythidaceae and make recommendations for a revised generic classification of the Bertholletia clade of Lecythidaceae.

English

Phytochemical investigation of the ethanol extract of leaves, twigs and fruit shell of Lecythis pisonis Camb. revealed the presence of squalene, a- e b-amyrin, lupeol, 3b-friedelinol, ursolic and oleanolic acids, (E)-phytol, sitosterol, stigmasterol, campesterol, quercetin-3-O-rutinoside and kaempferol-3-O-rustinoside. Structural elucidation was achieved using ultra violet (UV), nuclear magnetic resonance (NMR), and mass spectrometry. This is the first report of the occurrence of flavonoids, together with squalene, lupeol and campesterol in L. pisonis. The EtOH extract of the leaves showed high antioxidant activity, which can be associated in part with the high level of phenols and flavonoids. Key words: Lecythidaceae, Lecythis pisonis, triterpenes, flavonoids, antioxidant activity, total phenolic content, total flavonoid content.&nbsp;

SOBREVIVÊNCIA DE ESPECIES MADEIRÁVEIS EM SISTEMA AGROSSILVIPASTORIL EM SÃO DOMINGOS DO ARAGUAIA, PA

RESUMO: Os sistemas agrossilvipastoris têm sido uma alternativa indicada para reabilitação de áreas de pastagens degradadas. Objetivou-se avaliar a taxa de sobrevivência de mudas de espécies madeiráveis em ação-teste agrossilvipastoril implantada em unidade de produção familiar no assentamento “Belo Horizonte I”, em São Domingos do Araguaia-PA. Foram avaliadas durante um ano as espécies: ipê amarelo (Tabebuia serratifolia), ipê branco (T. roseo-alba), ipê roxo (Tabebuia cf. impetiginosa), oiti (Moquilea tomentosa), paricá (Schizolobium parahyba var. amazonicum), pata de vaca (Bauhinia longifolia), sapucaia (Lecythis pisonis) e sombreiro (Clitorea racemosa). As taxas de sobrevivência das espécies ipê amarelo, ipê branco, ipê roxo, paricá, oiti e sapucaia foram considerados satisfatórias sob as condições disponíveis da unidade de produção familiar estudada. PALAVRAS-CHAVES: agricultura familiar, sistema agroflorestal, taxa de sobrevivência.

Lecythis pisonis Cambess (sapucaia): seed morphometric traits and germinative response

This study aimed to test a possible correlation between germination and the variables weight and density of seeds of Lecythis pisonis (sapucaia), as well as raise issues which may contribute to know germination in this species. For this, we followed up the soaking and germination process of intact and scarified seeds of L. pisonis under controlled conditions (26°C, with average irradiance of 34.6 mmol.m-2.s-1 and in the dark). In another trial, seeds of sapucaia were individually characterized with regard to weight and density, and, then, sown in plastic boxes filled with soil and kept in a natural environment (average irradiance ±12.5 mmol.m-2.s-1 at 12:00 p.m.), with temperatures ranging from 15 to 32°C. The germination (shoot emergence) of each seed was followed up within the period from September 2009 to January 2010. The results suggest that the coat does not represent a restrictive factor for soaking and germination of sapucaia. Germination increased proportionally to the increase in seed density. Based on these results, we recommend the use of seeds with densities > 0.9 g.mL-1, in order to ensure a high percentage of seedlings.

Antinociceptive effect of Lecythis pisonis Camb. (Lecythidaceae) in models of acute pain in mice

Ethnopharmacological relevanceLecythis pisonis Camb., also known in Brazil as sapucaia, is used in folk medicine against pruritus, muscle pain and gastric ulcer. Aim of the studyTo investigate the antinociceptive effect of ethanol extract from Lecythis pisonis leaves (LPEE), fractions (hexane-LPHF, ether-LPEF and ethyl acetate-LPEAF) and mixture of triterpenes [ursolic and oleanolic acids (MT)] in mice. Materials and methodsLPEE and LPEF were evaluated on the acetic acid induced writhings and formalin, capsaicin and glutamate tests. In addition, MT was investigated on the writhings induced by acetic acid, capsaicin and glutamate tests. In the study of some possible mechanisms involved on the antinociceptive effect of LPEF, it was investigated the participation of opioid system, K+ATP channels and l-arginine-nitric oxide pathway. ResultsLPEE (12.5 and 25mg/kg, p.o.), LPEF and MT (6.25, 12.5 and 25mg/kg, p.o.) reduced the writhings in comparison to saline. LPEE (100mg/kg, p.o.) and LPEF (50mg/kg, p.o.) were effective in inhibiting both phases of formalin test. In capsaicin test, LPEE (100 and 200mg/kg, p.o.), LPEF (12.5–50mg/kg, p.o) and MT (6.25–25mg/kg, p.o.) showed a significant antinociceptive effect compared to the control. LPEE (25 and 50mg/kg, p.o.), LPEF (50 and 100mg/kg, p.o.) and MT (12.5 and 25mg/kg, p.o.) reduced the glutamate-evoked nociceptive response. Treatment with naloxone, l-arginine and glibenclamide reversed the effect of LPEF in glutamate test. ConclusionsThese results indicate the antinociceptive effect of Lecythis pisonis leaves and suggest that this effect may be related to opioid pathway, K+ATP channels, and l-arginine-nitric oxide modulation. Furthermore, these data support the ethnomedical use of this plant.

Chemical constituents of Lecythis pisonis and cytotoxic activity

The phytochemical investigation of the ethanol extract from leaves of Lecythis pisonis Cambess., Lecythidaceae, resulted in the isolation of seven triterpenes: α- and β-amyrin, uvaol and erythrodiol, ursolic and oleanolic acids and 3β-friedelinol, as well as a mixture of sitosterol and stigmasterol steroids and a diterpene (E)-phytol. The structures of these compounds were identified by 1H and 13C NMR spectral analysis and compared with literature data. The mixture of triterpenes ursolic and oleanolic acids isolated from the active ethereal fraction showed moderate cytotoxic activity. This paper describes for the first time the phytochemical and cytotoxic study of Lecythis pisonis' leaves.

Characterization and fatty acids profile of the oils from Amazon nuts and walnuts

PurposeThe purpose of this paper is to characterize oils extracted from Bertholletia excelsa, Lecythis pisonis, Dipteryx lacunifera, Carya illinoensis and Juglans regia, regarding their characterization and fatty acid profile.Design/methodology/approachThe oils were extracted from oilseeds by cold pressing and physico‐chemical characterization was performed by using standard methods for oils and fats. The oxidative stability and fatty acid profile also were determined.FindingsAccording to the results, the physico‐chemical properties of oils from nuts and walnuts were comparable to those of good quality conventional oils. The oil seeds are a good source of unsaturated fatty acids, especially oleic and linoleic acids.Research limitations/implicationsImplies the identification of fatty acid profile and physico‐chemical properties of oils extracted from nuts and walnuts, and to prevent certain types of diseases.Originality/valueThe paper identifies a new source of essential fatty acids extracted from oilseeds.

Fluorescence imaging of light acclimation of brazilian atlantic forest tree species

In the pursuit of knowledge on the biological behavior of Brazilian Atlantic Forest tree species, this study evaluated the susceptibility of the light-demanding species, Schinus terebinthifolia Raddi., Pseudobombax grandiflorum (Cav.) A. Robyns and Joannesia princeps Vell., and of the shade-tolerant species, Hymenaea courbaril L. var. stilbocarpa and Lecythis pisonis Camb, to photoinhibition and acclimation capacity. These species were first cultivated under two irradiance conditions, I20 (20% direct sunlight radiation) and I100 (all-sky or direct sunlight) and then transferred from I20 to I100. The effects of the sudden increase in light radiation intensity on photosynthetic activity were then evaluated through chlorophyll (Chl) fluorescence imaging, HPLC xanthophylls analysis, and cell membrane lipid peroxidation measurements. Light-demanding species were found to present a higher photochemical efficiency and higher acclimation capacity under high light irradiance than shade-tolerant species. The higher photoinhibition tolerance observed in light-demanding species was associated to their higher capacity for photochemical dissipation and dissipation of excess excitation energy via the xanthophyll cycle, leading to a lower ROS generation. The obtained results suggested that a knowledge of acclimation capacity, by means of Chl fluorescence imaging yields, is a useful indicator of species successional grouping.

Effects of Lecythis pisonis Camb. (Lecythidaceae) in a mouse model of pruritus

Ethnopharmacological relevanceLecythis pisonis Camb. (Lecythidaceae), is popularly known as “Sapucaia”. In traditional medicine, leaves are used for the treatment of pruritus. Aim of the studyThe present study is aimed to investigate the antipruritic effect of the ethanol extract from leaves of Lecythis pisonis (LPEE), fractions (hexane-LPHF, ether-LPEF and ethyl acetate-LPEAF) and mixture of triterpenes [ursolic and oleanolic acids (MT)] in mice and rats. Materials and methodsThe LPEE, LPHF, LPEF, LPEAF and MT were evaluated on scratching behavior induced by compound 48/80 in mice. In addition, LPEE, LPEF and MT were investigated on rat peritoneal mast cells degranulation induced by compound 48/80 (ex vivo study). The anti-inflammatory activity of LPEE and LPEF was investigated in rats using carrageenan-induced hind paw edema model. In the evaluation of the spontaneous motor activity, the LPEE was studied for its effect on spontaneous motor activity in an open-field test in mice. ResultsThe scratching behavior induced by compound 48/80 was significantly inhibited in mice pretreated with LPEE, LPHF, LPEF, LPEAF and MT. The suppressive effect of LPEE, LPEF and MT was only partially antagonized by naloxone. In addition, the compound 48/80-elicited degranulation of rat peritoneal mast cells was also markedly reduced in animals pretreated with LPEE, LPEF and MT. In the anti-inflammatory test, LPEE decreased the paw edema at the third hour after carrageenan (Carr) administration. Moreover, LPEF also was able to inhibit the oedematogenic response evoked by carr at all analysed time points. In the open-field test, LPEE-pretreated mice showed no impairment of spontaneous locomotion. Furthermore, the LPEE demonstrated no overt toxicity up to an oral dose of 2g/kg in an acute toxicity assay. ConclusionsThese results clearly indicate the antipruritic effects of Lecythis pisonis leaves and suggest that this effect may be related to a stabilizing action on mast cell membrane. Furthermore, these data support the traditional use of this plant against cutaneous pruritus.

Periconiella lecythidis sp. nov., the causal agent of a zonate leaf spot disease of the Brazilian tree Lecythis pisonis

A new defoliating disease was observed on 20-year-old trees of Lecythis pisonis in Vicosa, Minas Gerais, Brazil. The disease is characterized by forming one or more large zonate epiphyllous leaf spots with a light brown to white center, followed by two to three zones gradually varying from brown to red-brown. On the corresponding abaxial leaf surface, the spots are lighter with a profuse greenish gray coloration caused by the fungal colonies. In PDA culture, the fungus is characterized by a slow growth of the colonies with brown-violet pigmentation at the reverse. Inoculation tests carried out on healthy plants of L. pisonis confirmed the pathogenicity of this fungus. Periconiella lecythidis sp. nov., the causal agent of the zonate leaf spot disease of L. pisonis is described, illustrated, discussed and compared with the other species of the genus Periconiella. The intricate generic affinity within a complex of morphologically similar hyphomycete genera consisting of Metulocladosporiella, Parapericoniella, Penidiella and Periconiella is discussed in detail.

In vitro digestibility of globulins from sapucaia (Lecythis pisonis Camb.) nuts by mammalian digestive proteinases

Sapucaia (Lecythis pisonis Camb.) raw nuts collected from Brazil were analyzed to determine the proximate composition, amino acid profile of protein fractions, in vitro protein digestibility and antinutritional factors in order to evaluate their potential as a protein alimentary complement. The nuts contained adequate amounts of essential amino acids, fatty acids and minerals. In the present study, no hemagglutinating or inhibitory activities were observed in any of the samples investigated, indicating low or non-detectable levels of proteinase inhibitors or lectins in the samples. In vitro digestibility of in natura and heated nut globulins by mammalian digestive proteinases was carried out using trypsin + chymotrypsin + peptidase, with resulting mean values of approximately 70.30 and 71.35%, respectively. Taken together, the results suggest that sapucaia nuts may provide a new source of protein to use as a potential nutritional agent.

Carvão de resíduos de indústria madeireira de três espécies florestais exploradas no município de Paragominas, PA

Este trabalho avalia as principais propriedades do carvão de Manilkara amazonica (maçaranduba), Lecythis pisonis (sapucaia) e Piptadenia suaveolens (timborana), carbonizadas em sete e dez dias. O carvão foi produzido a partir de resíduos da serraria da Empresa Cikel Brasil Verde S. A., na Fazenda Rio Capim, em Paragominas. Foram utilizados 30 fornos de alvenaria, sendo dez fornos para cada espécie, dos quais cinco para carbonização em sete dias e cinco para dez dias. Após a carbonização, foram retiradas amostras de carvão produzido para estudo de rendimento gravimétrico e de propriedades físicas, químicas e mecânicas. Os principais resultados foram: o rendimento gravimétrico do carvão variou de 21,47 a 29,59 % (base úmida); a densidade média a granel, variou de 178,51 a 231,14 kg.m-3; a densidade média aparente foi de 0,38 a 0,53 g.cm-3; o poder calorífico foi de 23451 a 28830 kj.kg-1; o teor de materiais voláteis variou de 23,94 a 31,47%; cinzas de 0,7 a 2,5%; o carbono fixo de 68,29 a 74,49%; e a friabilidade de 12 a 32,6%. Constatou-se que a espécie é o principal fator que influencia nas propriedades do carvão. Através da análise das propriedades, não foi possível identificar qual espécie produziu o melhor carvão, uma vez que as três apresentaram algumas características consideradas boas, porém, sempre acompanhadas de outras inadequadas.

Classificação de sementes florestais quanto ao comportamento no armazenamento

Este trabalho teve como objetivos realizar um levantamento sobre a classificação de sementes de espécies florestais quanto ao comportamento durante o armazenamento e verificar a relação da classificação proposta com os grupos ecológicos das mesmas. Foram estudadas 39 espécies florestais presentes em remanescentes de matas ciliares na bacia do Alto e Médio Rio Grande, MG. A viabilidade das sementes e o grau de umidade foram obtidos para as sementes recém-beneficiadas; sementes recém-beneficiadas armazenadas em embalagem semipermeável a 5ºC durante 90 dias; e para sementes secas antes e após o armazenamento em embalagem impermeável sob temperaturas de 5ºC e -18ºC durante 90 dias. A análise estatística foi realizada comparando-se, por sobreposição, os intervalos de confiança das médias de porcentagem de germinação para cada espécie. As sementes das diferentes espécies foram classificadas como ortodoxas ou recalcitrantes durante o armazenamento. As sementes classificadas como ortodoxas são as que pertencem as espécies: Alchornea triplinervea, Anadenanthera colubrina, Aspidosperma cylindrocarpon, Aspidosperma polyneuron, Bowdichia virgilioides, Ceiba speciosa, Hymenaea courbaril, Lafoensia pacari, Lecythis pisonis, Lithraea molleoides, Maclura tinctoria, Myroxylon peruiferum, Myrsine umbellata, Rudgea viburnoides, Schinus terebinthifolius, Solanum granuloso-leprosum, Tabebuia crysotricha; e recalcitrantes as demais: Calophyllum brasiliense, Calyptranthes lucida, Cupania vernalis, Eugenia handroana, e Talauma ovata. O comportamento ortodoxo foi verificado para sementes de espécies pioneiras, clímax exigentes de luz para o crescimento da plântula e para espécies clímax tolerantes à sombra. Sementes classificadas como recalcitrantes foram encontradas para espécies clímax tolerantes à sombra e clímax exigentes de luz.

Lecythis pisonis Camb. nuts: oil characterization, fatty acids and minerals

The lipid content, the acid value, refractive and iodine indexes, the oil fatty acids profile as well as the concentration of inorganic elements contained in the Lecythis pisonis Camb. (Lecythidaceae) nuts were determined. The samples came from four different regions in São Paulo State, Brazil. The samples showed a high lipid content (34.2 to 61.3%) that presented a fatty acid profile, an iodine value and an index of refraction equivalent to the corn oils. Among the macronutrients, high levels of P (5.2 to 6.2 mg g −1) and Sn (69.1 to 77.0 μg g −1) were observed in all the nuts. High levels of Pb (3.3 to 3.8 μg g −1), Cu (2.9 to 3.3 μg g −1), Zn (2.6 to 3.8 μg g −1) and Mn (4.0 to 11.6 μg g −1) were found. It is evidence of a possible toxicity and it could compromise the use of the L. pisonis nuts for human consumption. ©