Intermediate molecular weight-fucosylated chondroitin sulfate from sea cucumber Cucumaria frondosa is a promising anticoagulant targeting intrinsic factor IXa.

Thromboembolic diseases pose a serious risk to human health worldwide. Fucosylated chondroitin sulfate (FCS) is reported to have good anticoagulant activity with a low bleeding risk. Molecular weight plays a significant role in the anticoagulant activity of FCS, and FCS smaller than octasaccharide in size has no anticoagulant activity. Therefore, identifying the best candidate for developing novel anticoagulant FCS drugs is crucial. Herein, native FCS was isolated from sea cucumber Cucumaria frondosa (FCScf) and depolymerized into a series of lower molecular weights (FCScfs). A comprehensive assessment of the in vitro anticoagulant activity and in vivo bleeding risk of FCScfs with different molecule weights demonstrated that 10 kDa FCScf (FCScf-10 K) had a greater intrinsic anticoagulant activity than low molecular weight heparin (LMWH) without any bleeding risk. Using molecular modeling combined with experimental validation, we revealed that FCScf-10 K can specifically inhibit the formation of the Xase complex by binding the negatively charged sulfate group of FCScf-10 K to the positively charged side chain of arginine residues on the specific surface of factor IXa. Thus, these data demonstrate that the intermediate molecular weight FCScf-10 K is a promising candidate for the development of novel anticoagulant drugs.

Sulfated Triterpene Glycosides from the Far Eastern Sea Cucumber Cucumaria djakonovi: Djakonoviosides C1, D1, E1, and F1; Cytotoxicity against Human Breast Cancer Cell Lines; Quantitative Structure-Activity Relationships.

Four new mono- and trisulfated triterpene penta- and tetraosides, djakonoviosides C1 (1), D1 (2), E1 (3), and F1 (4) were isolated from the Far Eastern sea cucumber Cucumaria djakonovi (Cucumariidae, Dendrochirotida), along with six known glycosides found earlier in other Cucumaria species. The structures of unreported compounds were established on the basis of extensive analysis of 1D and 2D NMR spectra as well as by HR-ESI-MS data. The set of compounds contains six different types of carbohydrate chains including two new ones. Thus, djakonovioside C1 (1) is characterized by xylose as the second residue, that was a branchpoint in the pentasaccharide chain. Meanwhile, only quinovose and rarely glucose have been found earlier in pentasaccharide chains branched at C-2 of the second sugar unit. Djakonovioside E1 (3) is characterized by a tetrasaccharide trisulfated chain, with glucose as the second residue. So, in the series of isolated glycosides, three types of sugars in the second position were presented: the most common, quinovose-in six compounds; glucose-in three substances; and the rare xylose-in one glycoside. The set of aglycones was composed of holostane- and non-holostane-type polycyclic systems; the latter comprised normal and reduced side chains. Noticeably, isokoreoside A (9), isolated from C. djakonovi, was a single glycoside having a 9(11)-double bond, indicating two oxidosqualenecyclases are operating in the process of the biosynthesis of aglycones. Some of the glycosides from C. djakonovi, which were characterized by pentasaccharide branched chains containing one to three sulfate groups, are chemotaxonomic features of the representatives of the genus Cucumaria. The assortment of sugar parts of Cucumaria's glycosides was broadened with previously undescribed penta- and tetrasaccharide moieties. The metabolic network of sugar parts and aglycones is constructed based on biogenetic relationships. The cytotoxic action of compounds 1-10, isolated from C. djakonovi, against human breast cancer cell lines was investigated along with the hemolytic activity. Erythrocytes were, as usual, more sensitive to the membranolytic action of the glycosides than cancer cells. The triple-negative breast cancer MDA-MB-231 cell line was more vulnerable to the action of glycosides in comparison with the other tested cancer cells, while the MCF-7 cell line was less susceptible to cytotoxic action. Djakonovioside E1 (3) demonstrated selective action against ER-positive MCF-7 and triple-negative MDA-MB-231 cell lines, while the toxic effect in relation to normal mammary epithelial cells (MCF-10A) was absent. Cucumarioside A2-5 (6) inhibited the formation and growth of colonies of cancer cells to 44% and tumor cell migration to 85% of the control. Quantitative structure-activity relationships (QSAR) were calculated on the basis of the correlational analysis of the physicochemical properties and structural features of the glycosidic molecules and their membranolytic activity. QSAR revealed the extremely complex nature of such relationships, but these calculations correlated well with the observed SAR.

Comparative physiological study of sea cucumbers from eastern waters of United States.

Sea cucumbers, belonging to the phylum Echinodermata, are known to possess valuable bioactive compounds that have medicinal properties. In several countries, such as Korea, China, and Japan, they are cultured in the aquaculture industries for food and medicinal purposes. Research has shown that different species of sea cucumbers each possesses unique medicinal values. As a result, we strive towards finding species with better health resilience in aquaculture system to be cultured for nutritional and medicinal purposes. In this paper, we compared the physiological and immunological parameters of three species of sea cucumbers, Cucumaria frondosa (C. frondosa), Isostychopus badionotus (I. badionotus), and Pentacta pygmaea (P. Pygmaea) from the waters of the eastern United States as they have not been studied extensively. Four different cells of sea cucumbers, phagocytic, red spherule, white spherule, and vibratile cells, that contribute to their immunity were counted. C. frondosa exhibited the highest concentrations of phagocytic cells, white spherule cells, and vibratile cells, compared to the two other species. Due to its high phagocytic cell concentration, the highest phagocytic capacity was seen in C. frondosa although it was not statistically significant. We also observed that C. frondosa had the highest total cell count and the highest concentration of coelomic protein among the three species. Lastly, C. frondosa possessed the highest lysozyme activity. Taken together, we concluded that C. frondosa is the best of the three species compared to be reared in the aquaculture systems for use in the food and biomedicine industries due to its immunological and physiological properties.

The Structural Characteristics and Bioactivity Stability of Cucumaria frondosa Intestines and Ovum Hydrolysates Obtained by Different Proteases.

The study aimed to investigate the effects of alcalase, papain, flavourzyme, and neutrase on the structural characteristics and bioactivity stability of Cucumaria frondosa intestines and ovum hydrolysates (CFHs). The findings revealed that flavourzyme exhibited the highest hydrolysis rate (51.88% ± 1.87%). At pH 2.0, the solubility of hydrolysate was the lowest across all treatments, while the solubility at other pH levels was over 60%. The primary structures of hydrolysates of different proteases were similar, whereas the surface hydrophobicity of hydrolysates was influenced by the types of proteases used. The hydrolysates produced by different proteases were also analyzed for their absorption peaks and antioxidant activity. The hydrolysates of flavourzyme had ß-fold absorption peaks (1637 cm-1), while the neutrase and papain hydrolysates had N-H bending vibrations. The tertiary structure of CFHs was unfolded by different proteases, exposing the aromatic amino acids and red-shifting of the λ-peak of the hydrolysate. The alcalase hydrolysates showed better antioxidant activity in vitro and better surface hydrophobicity than the other hydrolysates. The flavourzyme hydrolysates displayed excellent antioxidant stability and pancreatic lipase inhibitory activity during gastrointestinal digestion, indicating their potential use as antioxidants in the food and pharmaceutical industries.

Djakonoviosides A, A1, A2, B1-B4 - Triterpene Monosulfated Tetra- and Pentaosides from the Sea Cucumber Cucumaria djakonovi: The First Finding of a Hemiketal Fragment in the Aglycones; Activity against Human Breast Cancer Cell Lines.

Seven new monosulfated triterpene glycosides, djakonoviosides A (1), A1 (2), A2 (3), and B1-B4 (4-7), along with three known glycosides found earlier in the other Cucumaria species, namely okhotoside A1-1, cucumarioside A0-1, and frondoside D, have been isolated from the far eastern sea cucumber Cucumaria djakonovi (Cucumariidae, Dendrochirotida). The structures were established on the basis of extensive analysis of 1D and 2D NMR spectra and confirmed by HR-ESI-MS data. The compounds of groups A and B differ from each other in their carbohydrate chains, namely monosulfated tetrasaccharide chains are inherent to group A and pentasaccharide chains with one sulfate group, branched by C-2 Qui2, are characteristic of group B. The aglycones of djakonoviosides A2 (3), B2 (5), and B4 (7) are characterized by a unique structural feature, a 23,16-hemiketal fragment found first in the sea cucumbers' glycosides. The biosynthetic pathway of its formation is discussed. The set of aglycones of C. djakonovi glycosides was species specific because of the presence of new aglycones. At the same time, the finding in C. djakonovi of the known glycosides isolated earlier from the other species of Cucumaria, as well as the set of carbohydrate chains characteristic of the glycosides of all investigated representatives of the genus Cucumaria, demonstrated the significance of these glycosides as chemotaxonomic markers. The membranolytic actions of compounds 1-7 and known glycosides okhotoside A1-1, cucumarioside A0-1, and frondoside D, isolated from C. djakonovi against human cell lines, including erythrocytes and breast cancer cells (MCF-7, T-47D, and triple negative MDA-MB-231), as well as leukemia HL-60 and the embryonic kidney HEK-293 cell line, have been studied. Okhotoside A1-1 was the most active compound from the series because of the presence of a tetrasaccharide linear chain and holostane aglycone with a 7(8)-double bond and 16ß-O-acetoxy group, cucumarioside A0-1, having the same aglycone, was slightly less active because of the presence of branching xylose residue at C-2 Qui2. Generally, the activity of the djakonoviosides of group A was higher than that of the djakonoviosides of group B containing the same aglycones, indicating the significance of a linear chain containing four monosaccharide residues for the demonstration of membranolytic action by the glycosides. All the compounds containing hemiketal fragments, djakonovioside A2 (3), B2 (5), and B4 (7), were almost inactive. The most aggressive triple-negative MDA-MB-231 breast cancer cell line was the most sensitive to the glycosides action when compared with the other cancer cells. Okhotoside A1-1 and cucumarioside A0-1 demonstrated promising effects against MDA-MB-231 cells, significantly inhibiting the migration, as well as the formation and growth, of colonies.

Antioxidant and ACE-Inhibitory Activity of Protein Hydrolysates Produced from Atlantic Sea Cucumber (Cucumaria frondosa).

Atlantic sea cucumber is a benthic marine echinoderm found in Northwest Atlantic waters and is harvested mainly for its body wall. The body wall, along with internal organs and aquaphyrangeal bulb/flower, is a rich source of proteins, where the latter parts are often considered as processing discards. The objective of this research was to produce protein hydrolysates from sea cucumber tissues (body wall, flower, and internal organs) with bioactive properties associated with antioxidants, DNA and LDL cholesterol oxidation inhibition, and angiotensin-I-converting enzyme (ACE) inhibitory effects. The protein hydrolysates were prepared using food-grade commercial enzymes, namely Alcalase, Corolase, and Flavourzyme, individually and in combination, and found that the combination of enzymes exhibited stronger antioxidant potential than the individual enzymes, as well as their untreated counterparts. Similar trends were also observed for the DNA and LDL cholesterol oxidation inhibition and ACE-inhibitory properties of sea cucumber protein hydrolysates, mainly those that were prepared from the flower. Thus, the findings of this study revealed potential applications of sea cucumber-derived protein hydrolysates in functional foods, nutraceuticals, and dietary supplements, as well as natural therapeutics.

Copper exposure does not alter the ability of intertidal sea cucumber Cucumaria miniata to tolerate emersion during low tide.

Intertidal animals experience cycles of tidal emersion from water and are vulnerable to copper (Cu) exposure due to anthropogenic toxicant input into marine waters. Both emersion and Cu toxicity can cause damage to physiological processes like aerobic metabolism, ammonia excretion, and osmoregulation, but the interactions of the combination of these two stressors on marine invertebrates are understudied. Mixed effects of 96 h of low and high Cu exposure (20 and 200 µg/L) followed by 6 h of tidal emersion were evaluated on the intertidal sea cucumber Cucumaria miniata. The respiratory tree accumulated the highest concentrations of Cu, followed by the introvert retractor muscle, body wall, and coelomic fluid. Emersion affected accumulation of Cu, perhaps by inhibiting excretion. 200 µg/L of Cu increased lactate production in the respiratory tree, indicative of damaged aerobic metabolism. Cu diminished ammonia excretion, but emersion increased oxygen uptake and ammonia excretion upon re-immersion. The combination of the two stressors did not have any interactive effects on metabolism or ammonia excretion. Neither Cu exposure nor emersion altered ion (sodium, potassium, calcium, magnesium) content of the coelomic fluid. Overall, results of this study suggest that Cu exposure does not alter C. miniata's high tolerance to emersion, and some potential strategies that this species uses to overcome environmental stress are illuminated.

Phenolic profiles of Atlantic sea cucumber (Cucumaria frondosa) tentacles and their biological properties.

Sea cucumber tentacles (aquapharyngeal bulb) are a rich source of biologically active compounds, including phenolics, however they are mainly discarded as processing waste. This study evaluated free, esterified, and insoluble-bound phenolics of Atlantic sea cucumber (Cucumaria frondosa) tentacles and their antioxidant activity for the first time. Biological properties such as inhibitory activities against α-glucosidase, tyrosinase, and the formation of AGEs as well as LDL-cholesterol and DNA oxidation were investigated. The antioxidant activity of the phenolic extracts was also evaluated in a fish model system. In the UHPLC-QTOF-MS/MS analysis, 31 phenolic compounds, mainly phenolic acids and flavonoids, were identified and quantified. Among them, eight compounds were detected for the first time in any species of sea cucumber. The free phenolic fraction was the major form of phenolics, mainly protocatechuic acid, p-coumaric acid, gallic acid, ellagic acid, catechin, and quercetin, exhibiting strong antioxidant and biological activities. Fresh Atlantic salmon treated with sea cucumber phenolics delayed lipid oxidation as measured by the thiobarbituric acid reactive substances (TBARS) assay. Therefore, Atlantic sea cucumber tentacles may serve as a viable source of functional food ingredients with protective antioxidant properties.

Unusual Ether Lipids and Branched Chain Fatty Acids in Sea Cucumber (Cucumaria frondosa) Viscera and Their Seasonal Variation.

The sea cucumber, Cucumaria frondosa, is harvested primarily for its muscular bands and body wall. Development of a nutraceutical product based on lipid recovered from its viscera would give commercial value to the entire organism; however, such development requires knowledge of the lipid and fatty acid (FA) profiles of the viscera. Here, we describe the lipid and FA composition of viscera recovered from C. frondosa harvested in coastal waters in the northwest Atlantic, taking into account variation due to harvest season. We found highest lipid content at ~29% in winter, with diacylglyceryl ethers (DAGE) comprising ~55% of the total lipid mass and triacylglycerols (TAG), phospholipids (PL) and monoacylglycerol ethers (MAGE) at 5-25% each. The branched chain FA, 12-methyltetradecanoic acid (12-MTA), represented 42% of total FA mass in DAGE. In summer, lipid content was lower at 24% and TAG was the dominate lipid, with proportions more than double that found in winter (45% vs. 20%); DAGE in summer dropped to ~30% of total lipids. In TAG, 12-MTA was much lower than found in DAGE in winter, at only 10% but eicosapentaenoic acid (EPA) content was ~20%, which brought the total EPA% to 28% of total FA-the highest among all three seasons. There was little effect of season on MAGE or PL proportions. These data can help harvesters maximize catch efforts in terms of lipid yield and profile.

Functional expression and mutant analysis of thioredoxin-fused CEL-III, a hemolytic lectin from the marine invertebrate Cucumaria echinata.

CEL-III is a hemolytic lectin purified from the marine invertebrate Cucumaria echinata. New expression system of CEL-III was constructed, and the recombinant thioredoxin-fused CEL-III (Trx-CEL-III) showed strong hemolytic and carbohydrate-binding activity as same as authentic CEL-III. Mutation analysis of Trx-CEL-III suggested that carbohydrate binding to subdomain 1α and 2ß of CEL-III might be important for the hemolytic activity.

Effect of High-Pressure Processing (HPP) on Phenolics of North Atlantic Sea Cucumber (Cucumaria frondosa).

Sea cucumber contains a wide range of bioactive compounds, including phenolics. This study investigated the free, esterified, and insoluble-bound phenolics of sea cucumber body wall as affected by high-pressure processing (HPP) pretreatment. Sea cucumber body wall was subjected to HPP (200, 400, and 600 MPa for 5, 10, and 15 min), followed by the extraction of phenolics. The contents of total phenolics and antioxidant activity were monitored. Compared to untreated samples, those treated with HPP exhibited significantly higher total phenolics, flavonoids, and antioxidant activities. Treatment of 600 MPa for 10 min offered the optimal results. The highest amount of phenolics was observed in the free phenolic fraction, followed by esterified and insoluble-bound phenolic fractions. Moreover, phenolic extracts showed inhibitory effects against cupric ion-induced low-density lipoprotein (LDL)-cholesterol oxidation, peroxyl and hydroxyl radical-induced DNA scission, α-glucosidase activity, and formation of advanced glycation end products (AGEs). Ultra-high-performance liquid chromatography equipped with a quadrupole time of fight and mass spectrometer (UHPLC-QTOF-MS/MS) identified 20 phenolic compounds, mainly phenolic acids and flavonoids, from the body wall of this species for the first time. Thus, sea cucumber may lead to the production of a multitude of value-added products.

Depolymerization of a fucosylated chondroitin sulfate from Cucumaria japonica: Structure and activity of the product.

Fucosylated chondroitin sulfate CJ from the body wall of sea cucumber Cucumaria japonica was depolymerized by the treatment with H2O2 in the presence of Cu(OAc)2. The molecular weight of the polysaccharide was decreased from 32 kDa to 5 kDa. The product CJ-DP was shown to contain l-Fuc, d-GalNAc, d-GlcA, and sulfate in molar proportions of 0.96:0.90:1.00:5.4, which were quite similar to those of the parent polysaccharide CJ. The NMR analysis revealed that CJ-DP, like the parent polysaccharide CJ, consisted of both branched →4)-[3-O-α-l-Fuc]-ß-d-GlcA-(1 â†’ 3)-ß-d-GalNAc-(1→ and linear →4)-ß-d-GlcA-(1 â†’ 3)-ß-d-GalNAc-(1→ repeating blocks. Sulfate groups occupy O-4 and the majority of O-6 of GalNAc, as well as O-3 of GlcA, in linear blocks and different positions in Fuc branches. This result indicates that depolymerization practically does not diminish the amount of branches and sulfate groups in the product. Both polysaccharides CJ and CJ-DP demonstrated anticoagulant and hematopoiesis-stimulatory activities in vitro.

Fucoidans from Cucumaria frondosa ameliorate renal interstitial fibrosis via inhibition of the PI3K/Akt/NF-κB signaling pathway.

The effects of Cucumaria frondosa polysaccharides (CFPs) on renal interstitial fibrosis by regulating the phosphatidylinositol-3-hydroxykinase/protein kinase-B/nuclear factor-κB (PI3K/AKT/NF-κB) signaling pathway were investigated in vivo and in vitro in this research. The common unilateral urethral obstruction (UUO) model was used to examine the renoprotective effect and its mechanism in vivo. Compared to the UUO group, CFP administration could ameliorate renal function, inhibit inflammation and fibrosis, and reduce the deposition of the extracellular matrix and epithelial-mesenchymal transition. Mechanistic results indicated that CFPs could inhibit the expression of the total protein of PI3K and the conversion of the AKT and NF-κB p65 phosphorylated proteins, thereby inhibiting the transduction of the PI3K/AKT/NF-κB pathway. In addition, CFP treatment could improve inflammation and fibrosis in HK-2 cells induced by TGF-ß1, and its in vitro mechanism was also verified to inhibit the PI3K/Akt/NF-κB signaling pathway. Overall, these results showed that CFP could alleviate renal interstitial fibrosis related to the PI3K/AKT/NF-κB signaling pathway.

Inhibitory Effects of Sulfated Polysaccharides from the Sea Cucumber Cucumaria Frondosa against Aß40 Aggregation and Cytotoxicity.

Abnormal aggregation and deposition of Aß is one of the causative agents for Alzheimer's disease. The development of inhibitors for Aß aggregation has been considered a possible method to prevent and treat Alzheimer's disease. Edible sea cucumbers contain many bioactive molecules, including saponins, phospholipids, peptides, and polysaccharides. Herein, we report that polysaccharides extracted from sea cucumber Cucumaria frondosa could reduce the aggregation and cytotoxicity of Aß40. By utilizing multiple biochemical and biophysical instruments, we found that the polysaccharides could inhibit the aggregation of Aß40. A chemical kinetics analysis further suggested that the major inhibitory effects of the polysaccharides were achieved by disassembling mature fibrils, which in turn reduced the cytotoxicity of Aß. These results suggested that the polysaccharides extracted from sea cucumber could be used as an effective inhibitor for Aß.

Morphometrics and processing yield of Cucumaria frondosa (Holothuroidea) from the St. Lawrence Estuary, Canada.

Sea cucumber Cucumaria frondosa have highly variable whole body mass and length, and are usually sold to Asian markets as dried gutted body wall. Understanding the relation between size and yield of dry product is essential for resource conservation and for economic purposes. In this study, stock-specific mass and length recovery rates were estimated for C. frondosa captured by dredging or diving at various depths and seasons on the South shore of the St. Lawrence Estuary, along Gaspé Peninsula, and processed in a commercial plant. The processing yield in dry product mass per sea cucumber was more than 1.5 times larger for sea cucumbers collected at 26-47 m depth compared to those collected at 9-16 m depth. Within each strata, there was little variation in the processed body mass, seasonally or spatially. Recovery rates based on gutted mass for this stock (13.4─14.5%) varied little among depths and seasons, despite observed seasonal and bathymetric variation in reproductive status. In contrast, recovery rates based on whole body mass and length were highly variable both seasonally and spatially. Stress related to dredging or post-capture handling induced important variable body contraction and water content, leading to variation in body length, mass and shape of sea cucumbers having the same processed body mass. Gutted mass was the best metric to predict processed body mass and to estimate size whereas whole body length was the least reliable. New stock-specific information on variability of body mass, length, and recovery rates induced by capture, and on seasonal and bathymetric variation in reproductive status and processing yields will be used for the design of future stock assessment surveys, and for stock conservation.

Acute occupational exposure to holothurians (Cucumaria frondosa) resulting in irritating symptoms: About three cases.

Holothurians are marine invertebrates also known as sea cucumbers. They are used in fresh or dried forms in various cuisines around the world and have alleged medicinal properties. Consequently, sea cucumbers, notably the orange-footed sea cucumber (Cucumaria frondosa)are increasingly harvested from the environment or farmed via aquaculture. We report three cases of unusual occupational exposure to C. frondosa resulting in cutaneous, respiratory and ocular irritating symptoms. These symptoms occurred as sea cucumbers were cut on half and eviscerated manually, following a machine breakdown in a sea cucumber processing factory. Given the composition of holothurians, these symptoms probably resulted from the aerosolization of various holothurins, saponins secreted by sea cucumbers as mean of defense. Treatment was solely symptomatic and included decontamination of the skin and the eyes, inhaled glucocorticoids, anticholinergic agents and oral glucocorticoids. As the exposure resulted from a machine breakdown, no specific protective equipment was put in place. Employees were advised not to handle manuallythe sea cucumbers in case such a breakdown was to happen again.

From Coelomocytes to Colored Aggregates: Cellular Components and Processes Involved in the Immune Response of the Holothuroid Cucumaria frondosa.

AbstractWhile so-called brown bodies were first defined in the 1950s as colorful aggregates of cells in the general cavity of echinoderms and other marine benthic taxa, their distribution and role have not yet been fully clarified. This work characterized free coelomocytes and corresponding aggregates ("bodies") in the hydrovascular system and perivisceral coelom, as well as those attached on the membranes of the viscera, in the holothuroid Cucumaria frondosa. Responses to the presence of foreign particles were investigated, providing novel insights on the immune system. A total of eight coelomocyte cell types was detected, while aggregates were formed of three to six types of coelomocytes. Only red-colored aggregates were found in the hydrovascular system, whereas brown aggregates were confined to the perivisceral coelom. The encapsulation mechanism of foreign particles injected in the hydrovascular system was monitored. Particles were first gathered by phagocytes and vibratile, crystal, and morula cells into a whitish aggregate that was then covered by hemocytes, imparting a red color to the aggregates. After their transfer to the perivisceral coelom, aggregates became brown and were ultimately expelled through the anus. Finally, a range of stressors (i.e., harvesting method, presence of a predator, and physical injury) was found to increase the abundance of aggregates, thus highlighting the role of these bodies in the immune response of C. frondosa.

Features and Advantages of Supercritical CO2 Extraction of Sea Cucumber Cucumaria frondosa japonica Semper, 1868.

Extraction process of Cucumaria frondosa japonica Semper, 1868, which are subspecies of Cucumaria frondosa (Gunnerus, 1767), were studied. It was shown that supercritical carbon dioxide extraction of holothuria was more effective than conventional solvent extraction. Step-by-step extraction with carbon dioxide followed by supercritical extraction with the addition of a co-solvent of ethanol can almost double the yields of extracts of triterpene glycosides, styrenes and carotenoids. Moreover, the fraction of triterpene glycosides practically does not contain colored impurities, in contrast to traditional ethanol extraction. The obtained extracts by HPLC in combination with tandem mass spectrometry (HPLC-MS/MS) identified 15 triterpene glycosides, 18 styrene compounds and 14 carotenoids. Supercritical extraction made it possible to obtain extracts with yields superior to conventional hexane and alcohol extracts. Moreover, such an approach with the use of supercritical fluid extraction (SFE) and subsequent profiling of metabolites can help with the study of holothuria species that are not as well studied.

Sea Cucumber Sterol Alleviates the Lipid Accumulation in High-Fat-Fructose Diet Fed Mice.

The effect of marine-derived sea cucumber sterol (SS) with a special sulfate group on lipid accumulation remains unknown, although phytosterol has been proved to have many biological activities, including lowering blood cholesterol. The purpose of the present study is to investigate the alleviation of SS on lipid accumulation and the possible underlying mechanism using high-fat-fructose diet fed mice. Dietary administration with SS for 8 weeks reduced significantly the body weight gain and lipid levels in serum and liver. Especially, SS was superior to phytosterol in lowering lipid accumulation due to the great promotion of fatty acid ß-oxidation, the inhibition of cholesterol synthesis, and the acceleration of cholesterol efflux. The findings found that sea cucumber sterol exhibited a more significant effect than phytosterol on alleviating HFF-diet-induced lipid accumulation through regulating lipid and cholesterol metabolism, which might be attributed to the difference in the branch chain and sulfate group.

Galactose-Specific, Hemolytic Lectin CEL-III from Cucumaria echinata.

CEL-III is a Ca2+-dependent and galactose-specific lectin purified from the sea cucumber, Cucumaria echinata; it exhibits hemolytic and hemagglutinating activities. CEL-III consists of the following three distinct domains: two N-terminal carbohydrate-binding domains (1 and 2), which adopt ß-trefoil folds such as the B-chain of ricin and are members of the (QXW)3 motif family, and domain 3, an oligomerization domain. After binding to the cell surface carbohydrate chains through domains 1 and 2, domain 3 self-associates to form transmembrane pores composed of CEL-III heptamers, leading to cell lysis or death. In this chapter, the purification and carbohydrate-coated microplate binding assay of CEL-III are described.