Identification of triterpenoid saponins from Anemone rivularis var. flore-minore and study on their antitumor activities / 中国药房

OBJECTIVE To study the triterpenoid saponins from Anemone rivularis var. flore-minore and their antitumor activities. METHODS The n-butanol extract of 70％ ethanol extract from rhizome of the plant was separated. The triterpenoid saponins were separated and purified by normal silica gel column chromatography ，reversed phase ODS column chromatography ， Sephadex LH- 20 gel column chromatography and semi-preparation high performance liquid chromatography. The structures of these saponins were identified by spectral analysis （NMR and MS ）and physical and chemical properties. MTT assay was used to test the proliferation inhibitory activity of the compounds against five kinds of human tumor cells （HL-60 cells，A549 cells，HepG2 cells，HeLa cells and U 87MG cells ）. The apoptosis inducing effect of compound 7 on U 87MG cells was evaluated by flow cytometric Annexin V-FITC/PI staining test. RESULTS：Sixteen triterpenoid saponins were obtained and identified as 3 β-O-β-D-xylopyranosyl-（1→2）-α-L-arabinopyranosyl-oleanolic acid-28-O-α-L-rhamnopyranosyl- （1→4） -β-D-glucopyranosyl-（1→6）-β-D-glucopyranoside（1），3β-O-L-arabinopyranosyl oleanolic acid- 28-O-β-D-glucopyranoside（2），saponin B （3）， 163.com oleanolic acid- 3β-O-β-D-glucopyranosyl-（1→2）-α-L-arabino- pyranoside（4），HN-saponin F （5），clematoside S （6），prosapogenin CP 4（7），cussonside B （8），pulsatilla saponin C （9）， clemastanoside D （10），3 β-O-β-D-glucopyranosyl-（1→2）-β-L-arabinopyranosyl-hederagenin-28-O-β-D-glucopyranoside（11）， ciwujianoside C 3（12），ciwujianoside A 1（13），huzhangoside D （14），kalopanaxsaponin B （15）and hederacolchiside E （16）. Compounds 3，4，6-9 displayed inhibitory activities on the proliferation of tumor cells to different extent ，and compound 7 had the strongest activity ；compound 7 induced the apoptosis of U 87MG cell so as to inhibit the proliferation of cancer cells in a time-dependent manner. CONCLUSIONS The obtained 16 saponins are all identified as oleanolane-type ，among which compound 1 is a new compound. The monodesmosidic saponins ，the sugar chain of which attached at C- 3 and a free carboxyl at C- 28， possess stronger antitumor activity than others.

Diversity and antimicrobial activity of culturable fungi associated with sea anemone Anthopleura xanthogrammica

Background: The main objective of this study was to isolate fungi associated with Anthopleura xanthogrammica and measure their antimicrobial and enzymatic activities. A total of 93 fungal strains associated with A. xanthogrammica were isolated in this study, of which 32 isolates were identified using both morphological characteristics and internal transcribed spacer (ITS) sequence analysis. The antibacterial activities of 32 fungal isolates were tested against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Edwardsiella tarda, Vibrio harveyi, Fusarium oxysporum, and Pyricularia oryzae by agar diffusion assay. Extracellular hydrolytic enzyme activities of the fungal isolates were determined by agar diffusion assays. Enzyme activities were detected from clear halo size. Results: The isolated fungi belonged to 18 genera within 7 taxonomic orders of 1 phylum. The genera Aspergillaceae were the most diverse and common. The antimicrobial activities of 32 isolates were evaluated, and 19 (59.4%) of fungi isolate displayed unique antimicrobial activities. All fungal strains displayed at least one enzyme activity. The most common enzyme activities in the fungi isolates were amylase and protease, while the least common were pectinase and xylanase. Conclusions: This is first report on the sea anemone-derived fungi with antimicrobial and enzyme activities. Results indicated that sea anemone is a hot spot of fungal diversity and a rich resource of bioactive natural products.

Research progress of studies on chemical constituents and biologic activities of Anemone species / 中国中药杂志

Anemone is an important genus which was distributed widely and used to folk medicines in China. It is rich of pentacyclic triterpenoid saponins,and more than 100 kinds of pentacyclic triterpenoid saponins had been isolated and identified. Anemone has been used to treat punch injury and rheumatoid arthritis. This article reviews the latest research progress of Anemone decoction from two aspects: chemical constituents and pharmacological. It will provide reference for further research and development of Anemone.

Transcriptomic analysis in Anemone flaccida rhizomes reveals ancillary pathway for triterpene saponins biosynthesis and differential responsiveness to phytohormones / 中国天然药物

Anemone flaccida Fr. Schmidt is a perennial medicinal herb that contains pentacyclic triterpenoid saponins as the major bioactive constituents. In China, the rhizomes are used as treatments for a variety of ailments including arthritis. However, yields of the saponins are low, and little is known about the plant's genetic background or phytohormonal responsiveness. Using one-quarter of the 454 pyrosequencing information from the Roche GS FLX Titanium platform, we performed a transcriptomic analysis to identify 157 genes putatively encoding 26 enzymes involved in the synthesis of the bioactive compounds. It was revealed that there are two biosynthetic pathways of triterpene saponins in A. flaccida. One pathway depends on β-amyrin synthase and is similar to that found in other plants. The second, subsidiary ("backburner") pathway is catalyzed by camelliol C synthase and yields β-amyrin as minor byproduct. Both pathways used cytochrome P450-dependent monooxygenases (CYPs) and family 1 uridine diphosphate glycosyltransferases (UGTs) to modify the triterpenoid backbone. The expression of CYPs and UGTs were quite different in roots treated with the phytohormones methyl jasmonate, salicylic acid and indole-3-acetic acid. This study provides the first large-scale transcriptional dataset for the biosynthetic pathways of triterpene saponins and their phytohormonal responsiveness in the genus Anemone.

Identification of a pore-forming protein from sea anemone Anthopleura dowii Verrill (1869) venom by mass spectrometry

Background: Pore-forming proteins (PFP) are a class of toxins abundant in the venom of sea anemones. Owing to their ability to recognize and permeabilize cell membranes, pore-forming proteins have medical potential in cancer therapy or as biosensors. In the present study, we showed the partial purification and sequencing of a pore-forming protein from Anthopleura dowii Verrill (1869). 17. Methods: Cytolytic activity of A. dowii Verrill (1869) venom was determined via hemolysis assay in the erythrocytes of four mammals (sheep, goat, human and rabbit). The cytotoxic activity was analyzed in the human adherent lung carcinoma epithelial cells (A549) by the cytosolic lactate dehydrogenase (LDH) assay, and trypan blue staining. The venom was fractionated via ammonium sulfate precipitation gradient, dialysis, and ion exchange chromatography. The presence of a pore-forming protein in purified fractions was evaluated through hemolytic and cytotoxic assays, and the activity fraction was analyzed using the percent of osmotic protections after polyethylene glycol (PEG) treatment and mass spectrometry. 18. Results: The amount of protein at which the venom produced 50% hemolysis (HU50) was determined in hemolysis assays using erythrocytes from sheep (HU50 = 10.7 ± 0.2 µg), goat (HU50 = 13.2 ± 0.3 µg), rabbit (HU50 = 34.7 ± 0.5 µg), and human (HU50 = 25.6 ± 0.6 µg). The venom presented a cytotoxic effect in A549 cells and the protein amount present in the venom responsible for producing 50% death (IC50) was determined using a trypan blue cytotoxicity assay (1.84 ± 0.40 µg/mL). The loss of membrane integrity in the A549 cells caused by the venom was detected by the release of LDH in proportion to the amount of protein. The venom was fractionated; and the fraction with hemolytic and cytotoxic activities was analyzed by mass spectrometry. A pore-forming protein was identified. The cytotoxicity in the A549 cells produced by the fraction containing the pore-forming protein was osmotically protected by PEG-3350 Da molecular mass, which corroborated that the loss of integrity in the plasma membrane was produced via pore formation. 19. Conclusion: A. dowii Verrill (1869) venom contains a pore-forming protein suitable for designing new drugs for cancer therapy.(AU)

Review of Anemone raddeana Rhizome and its pharmacological effects / 中国结合医学杂志

The chemical compositions of Anemone raddeana Rhizome, a kind of traditional Chinese medicine, were reviewed, along with its bioactivity and pharmacological properties and method improvements of extracting and detecting triterpenoid saponins. A. raddeana Rhizome is used to treat neuralgia and rheumatism, and is rich in triterpenoid saponins, most of which are pentacyclic, with oleanane as the nucleus. So far, 37 triterpenoid saponins have been determined from the herb. Its reported bioactivity and pharmacological properties have been described as anticancerous, antimicrobial, anti-inflammatory, analgesic, antipyretic, anticonvulsive, antihistaminic, and sedative. It has also been used for the induction of the humoral immune response and treatment of liver fibrosis in chronic hepatitis. However, the herb also has hemolytic effects and can be toxic, which limits its clinical application. Further studies are needed on the pharmaceutical functions, mechanisms, and immunological responses to contribute to the herb's clinical applications.

Study on effect of different extraction methods on anti-inflammatory and analgesic activity of Anemone hupehensis / 重庆医学

Objective To investigate the effect of different extraction methods on anti-inflammatory and analgesic activity of Anemone hupehensis.Methods The different abstracts were prepared from the whole herb of Anemone hupehensis.The analgesic effect was observed by adopting the mouse torsion and electric heating plate method,and the anti-inflammatory activity was comprehensively evaluated by using the mouse ear tumefaction,toe tumefaction and tampon granulation tumefaction exprements.Results Compared with the blank model group,the anti-inflammatory action difference of low dose in the water layer parts of mouse ear tumefaction,toe tumefaction and tampon granulation tumefaction had no statistical significance(P＞0.05),and the extracting parts of rest doses all had significant anti-inflammatory and analgesic effect (P＜0.05).Ethyl acetate part had strongest activity in the electric heating plate experiment.N-butanol part had strongest activity in the ear tumefaction,toe tumefaction,tampon granulation tumefaction experiments and torsion method.Conclusion The whole herb of Anemone hupehensis has prominent anti-inflammatory and analgesic effect,and the ethyl acetate part E and N-butanol part are main effective parts.

Dose-effect Relationship of Yuning Ointment and its Decomposed Recipes on Acne in Mice / 中国药师

Objective: To study the dose-effect relationship of Yuning ointment and its decomposed recipes in the treatment of oleic acid induced acne in mice. Methods: Oleic acid was administrated to the back (2 cm ×2 cm) of the mice (once a day) for 21 days to induce acne. At d22, the gradient dosage of Anemone flaccida crude drug (1. 06-1 060. 23 mg?kg-1?d-1,k=3. 16), Yuning oint-ment without Anemone flaccida crude drug (4. 73-1 767. 75 mg?kg-1?d-1, k=3. 16) and Yuning ointment (2. 84-2 827. 28 mg?kg-1?d-1, k=3. 16) was respectively administrated to the back of mice for 14 days. The pathological changes of skin were observed by hematoxylin-eosin (HE) staining. The diameter of sebaceous glands and the ratio of follicular keratinization area were morphomet-rically analyzed. The serum levels of IL-1, IL-6 and TNF-α were detected by ELISA assay. The median effective dosages (ED50) of A-nemone flaccida in the three prescriptions were regressed by Prism 5. 01 software to determine the prescription dose-effect. Results: All the therapy groups were with significantly relieved pathological changes of sebaceous glands hypertrophy and follicular keratinization, and decreased serum levels of IL-1, IL-6 and TNF-α in a dose-dependent manner. The dose-response curves showed an "S" shape. A-mong the three therapy groups, the effect of Yuning ointment was the best. The ED50of Yuning ointment regressed by Anemone flaccida dose was 0. 28-fold for improving sebaceous glands hypertrophy, 0. 14-fold for inhibiting follicular keratinization, and 0. 15-, 0. 49-and 0. 24-fold for decreasing serum levels of IL-1, IL-6 and TNF-α. . Regressed by Yuning ointment without Anemone flaccida, the ED50of Yuning ointment was lower than Yuning ointment without Anemone flaccid in terms of improving pathological changes and inhibiting the secretion of cytokines. Conclusion: Yuning ointment can prevent and treat acne through regulating immune function. And the prescrip-tion compatibility can enhance the effects of Anemone flaccida.

Estudio bioquímico del veneno de la anémona de mar Phymactis papillosa (Actiniidae)

En este trabajo se ha estudiado bioquímicamente el veneno de Phymactis papillosa, colectadas en la bahía de Ancón. El veneno fue obtenido mediante shock hipotónico y luego se liofilizó. El análisis electroforético del veneno soluble mostró la presencia de 5 bandas proteicas con pesos moleculares entre 5 y 25.1 kDa. El veneno soluble fue fraccionado por cromatografía de filtración en una columna de Sephadex G-50, obteniéndose cuatro picos de proteína (I, II, III y IV). Tanto en el veneno soluble como en las fracciones colectadas se midió actividad de proteasa, fosfolipasa, hialuronidasa, fosfatasa ácida y fosfatasa alcalina; así como, actividad hemolítica y neurotóxica. Se encontró actividad proteolítica sobre caseína, en el veneno soluble y en los picos I y III. No se detectó actividad de fosfolipasa, hialuronidasa, fosfatasa ácida y fosfatasa alcalina. La actividad hemolítica, ensayada sobre eritrocitos humanos, se encontró en el veneno soluble y en el pico II. Finalmente, tanto el veneno soluble como el pico III mostraron ser neurotóxicos al ser inyectados en ratones albinos vía intraperitoneal. Se concluye que el veneno soluble de P. papillosa tiene actividad proteolítica, hemolítica y neurotóxica

In this work, the poison of Phymactis papillosa collected in Ancón bay has been studied biochemically. The venom was obtained by hypotonic shock and then lyophilized. Electrophoretic analysis of the soluble poison showed the presence of 5 protein bands with molecular weights between 5 and 25.1 kDa. The soluble venom was fractionated by filtration chromatography on a Sephadex G-50 column, yielding four protein peaks (I, II, III and IV). In the soluble venom and collected fractions was measured protease activity, phospholipase, hyaluronidase, acid phosphatase and alkaline phosphatase; as well as hemolytic and neurotoxic activity. Proteolytic activity on casein was found in the soluble venom and peaks I and III. Was not detected phospholipase activity, hyaluronidase, acid phosphatase and alkaline phosphatase. Hemolytic activity on human red cells tested, was found in the soluble venom and peak II. Finally, the soluble venom as the peak III showed be neurotoxic when injected into white mice intraperitoneally. It is concluded that the soluble venom of P. papillosa has proteolytic, hemolytic and neurotoxic activity

Study on the Extraction Technology of Phenolic Acids from Anemone altica / 中国药房

OBJECTIVE:To determine the content of phenolic acids from Anemone altica,and optimize its extraction technolo-gy. METHODS:HPLC was used to determine the contents of mono-ferulyl tartaric acid and ferulic acid from A. altica;using the total contents of 2 index components as index,volume fraction of extraction solvent,extraction solvent volume,extraction times and extraction time as factors,orthogonal test was used to optimize extraction technology,and verification test was conducted. RE-SULTS:The contents of mono-ferulyl tartaric acid and ferulic acid were 0.059%,0.0025%,respectively;the optimal extraction technology was as follow as 30% ethanol 600 mL added to 20 g medicinal material,extracted twice,90 min every time. In verifi-cation test,the average contents of 2 components in extract were 0.2971%(RSD=3.64%,n=3),0.0041%(RSD=5.11%,n=3). CONCLUSIONS:A method for contents determination of mono-ferulyl tartaric acid and ferulic acid from A. altica is estab-lished;optimized extraction technology is stable and feasible.

medicinal plants: ethnopharmacology, phytochemistry and biology

The Ranunculaceae genus(order Ranunculales), comprising more than 150 species, mostly herbs, has long been used in folk medicine and worldwide ethnomedicine. Various medicinal compounds have been found inplants, especially triterpenoid saponins, some of which have shown anti-cancer activities. Somecompounds and extracts display immunomodulatory, anti-inflammatory, antioxidant, and antimicrobial activities. More than 50 species have ethnopharmacological uses, which provide clues for modern drug discovery.compounds exert anticancer and other bioactivitiesmultiple pathways. However, a comprehensive review of themedicinal resources is lacking. We here summarize the ethnomedical knowledge and recent progress on the chemical and pharmacological diversity ofmedicinal plants, as well as the emerging molecular mechanisms and functions of these medicinal compounds. The phylogenetic relationships ofspecies were reconstructed based on nuclear ITS and chloroplast markers. The molecular phylogeny is largely congruent with the morphology-based classification. Commonly used medicinal herbs are distributed in each subgenus and section, and chemical and biological studies of more unexplored taxa are warranted. Gene expression profiling and relevant "omics" platforms could reveal differential effects of phytometabolites. Genomics, transcriptomics, proteomics, and metabolomics should be highlighted in deciphering novel therapeutic mechanisms and utilities ofphytometabolites.

Study on the HPLC Fingerprint of Anemone raddeana / 中国药房

OBJECTIVE:To establish the HPLC fingerprint for Anemone raddeana. METHODS:HPLC was performed on the column of Phemomenex Gemini C18 with mobile phase of 0.1%phosphoric acid-acetonitrile(gradient elution)at a flow rate of 1 ml/min,the detection wavelength was 206 nm,the column temperature was 30℃,and the injection volume was 20μl. With the refer-ence of raddeanin A,13 batches of A. raddeana were analyzed,chromatographic fingerprint similarity evaluation system software was conducted for similarity analysis,and SPSS 13.0 was conducted for cluster analysis. RESULTS:There were 11 common peaks in the 13 batches of A. raddeana with similarity of higher than 0.90. According to the verification,the fingerprint and control fin-gerprint shows good consistency. The drugs in Huadian,Jiaohe, Tiangang,Shulan,Tonghua and Fusong of Jilin and Shangzhi of Heilongjiang were regarded as category 1,and in Harbin,Yabuli town and Yimianpo of Heilongjiang,Qingyuan of Liaoning,Ji-nan of Shandong were category 2. CONCLUSIONS:The established fingerprint can provide reference for the identification and quality evaluation of A. raddeana.

Antimicrobial properties of sea anemone Anthopleura nigrescens from Pacific coast of Costa Rica

Objective: To evaluate the antimicrobial and antifungal activities of the aqueous and partitioned extract of sea anemone Anthopleura nigrescens (A. nigrescens). Methods: The sea anemone A. nigrescens was collected, minced, homogenized, lyophilized and then further partitioned with diethyl ether, acetone, ethanol and water. These fractions were evaluated for antimicrobial activity against bacterial and fungal pathogens. Results: Acetone extract was found to produce a pronounced inhibition of 7.0 mm against Proteus vulgaris and diethyl ether extract inhibited Pseudomonas aeruginosa with an inhibition zone of 6.5 mm. In antifungal activity, ethanol extract showed good activity against Botrytis cinerea, Trichoderma harzianum and Rhizopus oryzae compared with other strains. Acetone and ethanol extract of A. nigrescens showed activity against all of pathogens tested. Slight activity was observed in the water extract with inhibition zone of 1.5 mm. Conclusions: The present study revealed that sea anemone A. nigrescens may also contain some biologically active agents which have potential activity against pathogenic microorganisms.

Antimicrobial properties of sea anemone Anthopleura nigrescens from Pacific coast of Costa Rica

Objective: To evaluate the antimicrobial and antifungal activities of the aqueous and partitioned extract of sea anemone Anthopleura nigrescens (A. nigrescens). Methods: The sea anemone A. nigrescens was collected, minced, homogenized, lyophilized and then further partitioned with diethyl ether, acetone, ethanol and water. These fractions were evaluated for antimicrobial activity against bacterial and fungal pathogens. Results: Acetone extract was found to produce a pronounced inhibition of 7.0 mm against Proteus vulgaris and diethyl ether extract inhibited Pseudomonas aeruginosa with an inhibition zone of 6.5 mm. In antifungal activity, ethanol extract showed good activity against Botrytis cinerea, Trichoderma harzianum and Rhizopus oryzae compared with other strains. Acetone and ethanol extract of A. nigrescens showed activity against all of pathogens tested. Slight activity was observed in the water extract with inhibition zone of 1.5 mm. Conclusions: The present study revealed that sea anemone A. nigrescens may also contain some biologically active agents which have potential activity against pathogenic microorganisms.

Phenolic acid compounds from anemone altaicae rhizoma / 中国药学杂志

OBJECTIVE: To study the chemical constituents in the roots of Anemone altaica. METHODS: The isolation and purification of the compounds were performed by chromatography on Diaion HP-20, Sephadex LH-20, and silica gel, combined with preparation liquid chromatography. Their structures were determined by comparison of their physicochemical characteristics and spectral data with literatures. RESULTS: Eleven compounds were obtained, and their structures were identified as methyl feruloyl-tartarate A (I), methyl feruloyl-tartarate B (II), ethyl feruloyl-tartarate B (III), mono-methyl feruloyl-lactate (IV), dimethyl feruloyl-lactate (V), vanillic acid-4-O-β-D-glucopyranoside (VI), p-hydroxybenzoic acid glucoside (VII), methyl chlorogenate (VIII), 3-O-fernloylquini acid (IX), 5-O-feruloyl-3-O-(β-D-glucopyranosyl)-2-deoxy-D-ribono-γ-lactone (X), and carboxymethyl isoferulate (XI). CONCLUSION: Compounds I-V are new compounds, and are speculated to be a man-made products. Compounds VI-XI are isolated from the plants of anemone genus for the first time.

Chemical constituents from rhizoma of Anemone altaica / 中草药

Objective: To study the chemical constituents in the rhizoma of Anemone altaica. Methods: The isolation and purification of the compounds were performed by Diaion HP-20 macroporous resin, Sephadex LH-20, and silica gel column chromatography, and their structures were determined by comparing their physicochemical characters and spectral data with literatures. Results: Fourteen compounds were obtained, and their structures were identified, including six phenolic compounds: ferulic acid (1), mono-feruloyl-tartaric acid (2), chlorogenic acid (3), glucosyringic acid (4), feruloyl-6'-O-α-D-glucopyranoside (5), and feruloyl-6'-O-β-D-glucopyranoside (6); two lignans: (+)-isolariciresino-1-9-O-β-D-glucopyranoside (7) and (+)-pinoresinol-4-O-β-D-glucopyranoside (8); one courmarin: esculetin (9); and five nitrogen-containing compounds: 2, 3, 4, 9-tetrahydro-1H-pyridine pyrido [3, 4-b] indole-3-carboxylic acid (10), phenylalanine (11), adenine (12), thymidine (13), and adenosine (14). Conclusion: Compounds 2-13 are isolated from the plants of the genus Anemone L. for the first time.

Identification of saponins in rhizomes of Anemone davidii by UPLC/Q-TOF-MS/MS / 中草药

Objective: To identify the saponins in the rhizomes of Anemone davidii by ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UFLC/Q-TOF-MS/MS). Methods: The separation was performed on UPLC Welch C18 column (100 mm × 2.1 mm, 1.7 μm), with a mobile phase using 0.1% acetonitrile (A) and water containing 0.1% formic acid (B) for gradient elution. Q-TOF/MS and electrospray ion (ESI) source were applied for the analysis under the negative ion mode, and the running time was 40.25 min. Using target compound screening method, the structures of monitored chemical constituents were identified by retention time, exact relative molecular mass, and cleavage fragments of MS/MS. Results: Fifty-two triterpenoids were separated and identified from the methanol extract of A. davidii, 47 triterpenoids were identified for the first time from A. davidii among which nine pairs of structural isomers were included. Conclusion: UPLC/Q-TOF-MS/MS method can identify the main chemical constituents from A. davidii rapidly and accurately, which develops a new strategy for identification of the chemical constituents in A. davidii.

Ocurrence of the sea anemone Telmatactis panamensis (Verrill, 1869) (Cnidaria: Anthozoa: Actiniaria) at Isla del Coco National Park, Costa Rica / Ocurrencia de la anémona de mar Telmatactis panamensis (Verrill, 1869) (Cnidaria: Anthozoa: Actiniaria) en el Parque Nacional Isla del Coco, Costa Rica

The sea anemone fauna of Isla del Coco National Park (also known as Cocos Island Nacional Park), Pacific Costa Rica is poorly known. In the present work we report the first occurrence of the species Telmatactis panamensis. Individuals of this sea anemone (n=24) were collected at Chatham Bay intertidal and at 15m depth in Punta Ulloa, in both cases attached to rocks; during the expedition UCR-UNA-COCO-I in April 2010. We provide photographs of live individuals, external anatomy and an inventory of cnidae of the studied specimens. Possibly this species is extended to greater depth as observed by other authors in the Galápagos Islands.

La fauna de anémonas de mar es prácticamente desconocida para el Parque Nacional Isla del Coco (Costa Rica). En el presente trabajo se reporta por primera vez la presencia de la especie Telmatactis panamensis. Individuos de esta anémona de mar fueron colectados en el intermareal de Bahía Chatham y a 15m de profundidad en Punta Ulloa, en ambos casos adheridas a rocas; durante la expedición UCR-UNA-COCO-I en Abril de 2010. Se proveen fotografías de ejemplares vivos, datos de su anatomía externa y un inventario del cnidae de los especímenes estudiados. Posiblemente esta especie se extienda a mayor profundidad, tal como fue observado por otros autores para ejemplares de las Islas Galápagos.

Differential susceptibilities of human lung, breast and skin cancer cell lines to killing by five sea anemone venoms

Although sea anemones are well known for being rich sources of toxins, including cytolysins and neurotoxins, their venoms and toxins have been poorly studied. In the present study, the venoms from five sea anemones (Heteractis crispa, Heteractis magnifica, Heteractis malu, Cryptodendrum adhaesivum and Entacmaea quadricolor) were obtained by the milking technique, and the potential of these venoms to kill cancer cells was tested on three cell lines (A549 lung cancer, T47D breast cancer and A431 skin cancer). The total protein level in the crude extract was determined by the bicinchoninic acid (BCA) protein assay. The cytotoxicity on different cell lines was assayed using the 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide (MTT) assay which measures survival based on the detection of mitochondrial activity and by the crystal violet assay, which measures survival based on the ability of cells to remain adherent to microplates. The results indicate that the sea anemone venom is cytotoxic to human cancer cells. The A549 cell line was the most sensitive of the cell lines tested with a significant reduction in viability observed at 40 µg/mL. H. malu, C. adhaesivum and E. quadricolor had a significant inhibitory effect on A431 cells. Furthermore, H. malu and C. adhaesivum had a significant inhibitory effect on T47D cell line at 40 µg/mL. In conclusion, the sea anemone venoms tested have the potential to be developed as anticancer agents.(AU)

Biological activity of sea anemone proteins: II. Cytolysis and cell line toxicity.

Potent cytolytic activity was exhibited by proteins extracted from three sea anemones viz. Heteractis magnifica, Stichodactyla haddoni and Paracodylactis sinensis by affecting the red blood corpuscles (RBC) and the mouse fibroblast cell line (L929) and leukemia cell line (P388). Crude toxin of all the three anemone species induced spontaneous hemolysis of chicken, goat and human erythrocytes. The crude toxin of H. magnifica (0.98 mg/ml) elicited hemolysis at levels of 4096, 512 and 4096 HU (hemolytic unit) in chicken, goat and human erythrocytes respectively. Subsequently, the crude toxin of S.haddoni (0.82 mg/ml) exhibited a hemolytic activity of 256, 128 and 512 HU and that of P. sinensis (0.60 mg/ml) had a hemolytic activity of 128, 4096 and 512 HU. Most of the partially purified proteins of these anemones also exhibited the activity against the three different erythrocytes. The viability of L929 and P388 was adversely affected on adding the crude toxins. The symptoms of toxicity shown by the cells were rounding, lysis and detachment from the substratum. These effects were the least in S. haddoni, as compared to those the crude toxins of the other two species. Inhibition of growth of L929 exhibited by the toxin of the three species ranged between 61.08 and 93.38%. Similarly, inhibition of the growth of P388 ranged between 51.32 and 86.16%. The present investigation reveal the cytotoxic nature of anemone toxins.