Body Wall Extracts Research Articles

Comprehensive metabolomics of Philippine Stichopus cf. horrens reveals diverse classes of valuable small molecules for biomedical applications.

Stichopus cf. horrens is an economically important sea cucumber species in Southeast Asia due to their presumed nutritional and medicinal benefits. However, compared to other sea cucumbers such as Apostichopus japonicus, there are no biochemical studies on which compounds contribute to the purported bioactivities of S. cf. horrens. To address this, a high-throughput characterization of the global metabolite profile of the species was performed through LC-MS/MS experiments and utilizing open-access platforms such as GNPS, XCMS, and metaboAnalyst. Bioinformatics-based molecular networking and chemometrics revealed the abundance of phospholipids such as phosphatidylcholines (PCs), phosphatidylethanolamines (PEs), phosphatidylinositols (PIs), and phosphatidylserines (PSs) in the crude samples. Body wall extracts were observed to have higher levels of structural, diacylated PCs, while the viscera have higher relative abundance of single-tail PCs and PEs that could be involved in digestion via nutrient absorption and transport for sea cucumbers. PEs and sphingolipids could also be implicated in the ecological response and morphological transformations of S. cf. horrens in the presence of predatory and other environmental stress. Interestingly, terpenoid glycosides and saponins with reported anti-cancer benefits were significantly localized in the body wall. The sulfated alkanes and sterols present in S. cf. horrens bear similarity to known kairomones and other signaling molecules. All in all, the results provide a baseline metabolomic profile of S. cf. horrens that may further be used for comparative and exploratory studies and suggest the untapped potential of S. cf. horrens as a source of bioactive molecules.

Antibacterial potential of Luidia clathrata (sea star) tissue extracts against selected pathogenic bacteria.

As resistance to traditional antibiotics has become a major issue, it is essential to explore natural sources for new antimicrobial agents. The marine environment offers a variety of natural bioactive compounds. In this study, we examined the antibacterial potential of Luidia clathrata, a tropical sea star species. The experiment was conducted against both gram-positive (Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Bacillus cereus and Mycobacterium smegmatis) and gram-negative (Proteus mirabilis, Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) bacteria using disk diffusion method. Specifically, we extracted the body wall and gonad using methanol, ethyl acetate, and hexane. Our findings show that the body wall extract using ethyl acetate (1.78μg/ml) was particularly effective against all tested pathogens, while the gonad extract (0.107μg/ml) showed activity against six out of ten selected pathogens. This is a crucial and new discovery that suggests L. clathrata may be a useful source for discovering antibiotics and more research is required to pinpoint and comprehend the active ingredients.

Natural Extracts as Eco-Friendly Larvicides against Aedes aegypti Mosquito, Vector of Dengue Fever Virus in Jeddah Governorate

Severe human diseases are spread by mosquitoes, causing millions of deaths every year. Many well-known and severe problems have been caused by the indiscriminate use of synthetic chemical insecticides, such as the residual insecticides for humans and environment and high operating cost in addition to the possibility of developing insect resistance.The larvicidal and delayed effects of the body wall extract of Holothuria scabra and leaves extract of Acalypha fruticosa against 4th instar larvae of mosquito, Aedes aegypti were evaluated. Ethanolic extract of H. scabra recorded more larvicidal efficiency(LC50, 79.31 ppm) than A. fruticosa leaves extract (152.86 ppm) by about 1.93 folds.Morphological features showed abnormalities on the larval and pupal stages with H. scabra and to less extent with A. fruticosa. Therefore, it is possible to build on the results of this study to use these two extracts to control of A. aegypti mosquitoes and in line with recent trends in adopting combat methods that are safe on humans and the environment.

Anticancer Activity of <i>Holothuria atra</i> Extracts on the Triple-Negative Breast Cancer Cell Line MDA-MB-231

Holothuria atra (HA), black sea cucumber, has been widely used as a traditional medicine in several Asian countries. A previous study showed the potent anticancer effects of HA extracts on cell proliferation and apoptosis in various types of cancer cell lines, including breast cancer cells. However, the molecular mechanism by which HA extracts exert anticancer effects on the triple-negative breast cancer cell line MDA-MB-231 remains unclear and poorly understood. In the present study, we assessed the inhibitory effects of body wall (BW) and visceral (VS) extracts of HA on proliferation, adhesion, migration, invasion, and tube formation in an in vitro model of the highly metastatic triple-negative breast cancer cell line MDA-MB-231. BW and VS extracts were prepared by the ethyl acetate extraction process. Anticancer properties, including the inhibition of proliferation, adhesion, migration, invasion, and tube formation, were examined by MTT assays, cell adhesion assays, wound healing migration assays, Matrigel invasion assays, and tube formation assays, respectively. The protein expression levels of Bax, Bcl-2, Caspase-3, Akt, matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-9 (MMP-9) were examined by Western blot analysis. Our findings demonstrated that BW extract treatment exhibited significantly greater inhibitory effects on MDA-MB-231 cell proliferation in a time- and dose-dependent manner than VS extract treatment. Moreover, dose-dependent suppression of MDA-MB-231 cell adhesion, migration, invasion, and tube formation was observed in the BW extract-treated group. The inhibition of MDA-MB-231 cells invasion in the BW extract-treated group was achieved by suppressing the levels of MMP-2 and MMP-9 through the Akt/MMP pathway. MDA-MB-231 cell apoptosis was promoted after treatment with BW extract by downregulating Bcl-2 levels and upregulating Bax and Caspase-3 expression through Akt/Bad signaling. Our study revealed the anticancer effects of BW extracts of HA. This study provides a potential therapeutic strategy using beneficial HA components for treating patients with breast cancer. Funding Information: This research was funded by Mahidol University and the Thailand Research Fund (Grant IRG5780011). Declaration of Interests: The authors declare no conflict of interest.

The Sea Cucumber Body Wall Extract Promoted in Vitro Maturation of NMRI Mice Follicles at Germinal Vesicle Stage

Background: IVM (in vitro maturation) appear to be certified high pregnancy outcome. Therefore, attempt to find an appropriate culture system using natural products for increasing developmental competencies fascinating. Objectives: This experiment is aimed to evaluate the effect of sea cucumber methanol extract on in vitro maturation of immature mouse oocyte. Methods: In this experimental study, Preantral follicles at germinal vesicle stage were collected from 24 - 26 days NMRI female mouse ovaries and transferred to IVM medium supplemented with different concentration of sea cucumber methanol extract (5, 10, 20, 30 µg/mL). Antioxidant capacity of sea cucumber extract was evaluated using DPPH (2, 2-diphenyl-1-picrylhydrazyl) assay. Further, oocyte maturation and TNF-α expression were recorded till day 10th. Data were analyzed by one-way analysis of variance (ANOVA) through SPSS 16. Results: The percentage of arrested oocyte at germinal vesicle stage in the control group was significantly (P < 0.05) higher than treated group (20 µg/mL). There were significant (P < 0.01) differences in percentage of maturated oocyte to metaphase II (MII) stage between treated and control group. DPPH radical scavenging capacity and reduction of TNF-α expression in treated oocyte demonstrated that sea cucumber increased rate of oocyte maturation. In addition, the treated oocyte (20 µg/mL) achieved the highest percentage of MII stage. Conclusions: It was concluded that the sea cucumber extract with optimum concentration can improve oocyte maturation. Sea cucumber extract treatment can be suggested as a novel therapeutic strategy to be used in infertility modality in the future.

Extract of the sea cucumber, Holothuria scabra, induces apoptosis in human glioblastoma cell lines

Background: Glioblastoma is the most aggressive primary brain tumor resistant to conventional treatment, and has a frequent recurrence. Holothuria scabra is the most widely consumed sea cucumber in Thailand and Asian countries. Sea cucumber extracts have valuable bioactive ingredients that are utilized in Chinese traditional medicine for several therapeutic measures including anti-cancer. In this study, we aimed to test the anticancer activity of H. scabra body wall extract in ethyl acetate fraction against two human glioblastoma cell lines A172 and U87MG.Material and methods: The anticancer effect of the ethyl acetate fraction of body wall extracts from H. scabra (HSBWEA) was tested against two human glioblastoma cell lines A172 and U87MG to determine cell viability, apoptotic cell analysis, mitochondrial membrane potentials, nuclear condensation and expression of apoptotic molecules. The bioactive compound in the extract was characterized by LC-MS/MS.Results: The extract exhibited strong cytotoxic effect to A172 and U87MG cells with IC50 4.23 and 4.46 mg/ml, respectively. The extract also induced both early and late stages of apoptosis, loss of mitochondrial membrane potentials and nuclear condensation and fragmentation. After treatment with the extract for 24 h, the cells expressed increasing amounts of a pro-apoptotic Bax and caspase-3 while an anti-apoptotic Bcl-2 was decreased in a concentration-dependent manner. The major bioactive components in this extract are triterpene glycosides similar to those described previously in H. scabra as well as in other related sea cucumber species.Conclusions: The crude extract from the body wall of H. scabra exhibited anticancer activity against human glioblastoma cells through the mitochondrial-mediated apoptotic pathway, and the compounds in the extract might be a novel candidate for anti-glioblastoma.

Antiseptic effect of sea cucumber (Holothuria atra) against multi-organ failure induced by sepsis: Molecular and histopathological study.

Sepsis is a systemic inflammatory response to infection and severe sepsis patients can develop acute lung and liver injury. The aim of the present study was to evaluate the efficacy of Holothuria atra methanolic body wall extract (HaE), as an antioxidant and anti-inflammatory agent against induced sepsis in a cecal ligation and puncture (CLP) rat model. In total, 30 males albino rats were divided into three groups (n=10 each) as follows: Sham (Sh), which was used as negative control; sepsis (Se), which was used as a positive control and was subjected to CLP surgery; and Ho, which was subjected to CLP and fed with 200 mg/kg (body weight) of HaE, once daily for 7 days. Subsequently, the expression of various genes was detected by polymerase chain reaction, while liver and lung tissues were examined by immunohistochemistry. The expression of Caspase-3 was significantly reduced in liver and lung tissues in the Ho group, while the expression levels of Gsta2, Cat and Sod1 genes were slightly reduced in the Ho group, when compared with the Se group. In addition, expression levels of tumor necrosis factor, interferon-γ, liver interleukin (IL)1b and lung IL1a were reduced in the Ho group compared with the Se group. Furthermore, histopathological changes were observed in liver tissues of the Se group, including congestion of hepatoportal blood vessel and focal hepatic necrosis, while lung tissues showed marked edema, hemorrhage and alveolar septal thickening. The Ho group showed apparent normal hepatic parenchyma and slight interstitial pneumonia. Immunohistochemical staining of caspase-3 in liver and lung tissues showed no expression in the Sh group, strong expression in the Se group and moderate expression in the Ho group. In conclusion, HaE demonstrated beneficial effect against induced sepsis, which may be attributed to its antioxidant and antiapoptotic activities.

Antitumor and antifungal activities of organic extracts of seacucumber Holothuria atra from the southeast coast of India

In phylum Echinodermata, the family Holothuridae is distinguished by its capacity of bioactive compounds. Sea cucumber Holothuria atra is commonly known as the lollyfish. The antifungal activity was detected using agar well diffusion method against the various fungal strains such as Trichoderma viride, Aspergillus niger, Aspergillus flavis, Candida albicans and Penicillium chrysogenum. Relatively high antifungal activity was seen against Candida albicans at 100 μL−1 concentration of extracts. Zone of inhibition was measured at 18 mm of diameter. The anti-tumor activities were detected against the Vero and Hep2 cell lines using MTT assay. The cells were treated with H. atra extract at concentrations 0.078–10mg mL−1. The extract showed high proliferative activity against the Hep2 cells. The body wall extracts of sea cucumber (H. atra) showed effective antifungal and antitumor activities. All these findings suggest that the extracts could be used for the development of drugs.

Antibacterial activity of the body wall extracts of sea cucumber (Invertebrata; Echinodermata) on infectious oral streptococci.

Abstract Background: The present study was carried out to test the antibacterial effect of the body wall of the sea cucumber Holothuria leucospilota Brandt on Streptococcus mutans and Streptococcus salivarius. Methods: After sampling sea cucumbers from the Persian Gulf, different extractions were prepared. Then, aqueous, phosphate-buffered saline (PBS, pH 7.8), chloroform, hexane and methanolic extracts from sea cucumber body wall were screened for antibacterial activity against pathogenic bacteria S. mutans and S. salivarius using the disk diffusion method. Results: The PBS extract did not show any antibacterial or inhibitory activity; the chloroform extract, however, demonstrated high levels of antibacterial activity against S. salivarius while exhibiting low levels of activity against S. mutans. The hexane and methanolic extracts were found to show no antibacterial activity against S. mutans, but exhibited antibacterial activity against S. salivarius. Conclusions: In conclusion, the results demonstrated the possibility of utilizing sea cucumbers as a cheap source of potential antibacterial agents and for treating odontogenic diseases.

A NOVEL BIOFOULING ASSAY USING COMPUTER‐ASSISTED MOTION ANALYSIS OF HINCKSIA IRREGULARIS SPORE SWIMMING

Computer‐assisted motion analysis of the swimming behavior of spores of the brown alga Hincksia irregularis was used in a novel antifouling bioassay. Newly released spores were exposed to water soluble compounds of organic and aqueous body wall extracts of two sympatric sea stars, Astropecten articulatus and Luidia clathrata. Changes in the swimming behavior of H. irregularis spores were quantified by changes in speed (decrease at extract exposure) and rate of change of direction (increase at extract exposure). Sea star extracts were tested at natural concentrations (mg extract/g wet weight) and at approximate 10‐, 100‐, and 1000‐fold dilution. Significant effects (p ≤ 0.05) were observed at 1000‐fold dilution of the natural concentration in both extracts of A. articulatus, and at 100‐ and 10‐fold dilution of the aqueous and organic extract, respectively, in L. clathrata. Germination of spores after short time exposure (30 min) to sea star body wall extracts indicates that compounds were not toxic but affected spore mobility, a factor that plays an important role during spore settlement. We suggest that this assay is very useful for bioassay‐guided fractionation of extracts because it is very sensitive and requires only small amounts of extracts or isolated compounds.

UNPALATABLE COMPOUNDS IN THE MARINE GASTROPOD Dolabella auricularia: DISTRIBUTION AND EFFECT OF DIET

Sea hares are a rich source of novel secondary metabolites, most of which are derived from their algal diet, but the natural function(s) of these metabolites are largely unknown. We used field and laboratory assays to measure the palatability of extracts from the tissues, ink, and eggs of Dolabella auricularia. Digestive-gland extracts contained a wide variety of secondary metabolites, including the red algal compound prepacifinol epoxide and its derivative johnstonol, and they were unpalatable to reef fishes. Skin extracts were moderately unpalatable, but our bioassay-guided fractionation led us to (–)-7-dehydrocholesterol, rather than to an algal secondary metabolite. Ink extracts were consistently unpalatable to reef fishes only at high concentrations, suggesting either that ink must be concentrated to deter predators, that unpalatable components of ink rapidly decompose, or that ink has other functions. Unpalatability of ink was traced to a purple fraction, consistent with the hypothesis that the active compound is aplysioviolin, a known ink constituent modified from a red algal pigment. Egg extracts were moderately unpalatable; however, we could not trace this activity to any algal-derived secondary metabolite. Body-wall extract was highly palatable. Our results suggest that dietary-derived secondary metabolites play a role in chemical defense of D. auricularia via the ink, but are not responsible for unpalatability of skin or eggs. Accumulation of dietary-derived metabolites in the digestive gland may occur to detoxify a chemically rich diet, rather than or in addition to deterring predators.

Structural and chemical defenses of echinoderms from the northern Gulf of Mexico

The feeding deterrent effects of echinoderm body-wall tissues and ethanolic extracts containing mid-polarity compounds were evaluated utilizing generalist fish and crabs as model predators. The body-wall tissues of the echinoderms examined ranged 10-fold from 0.9–9.4 mm in thickness, and four and a half-fold in level of mineralization (17.8–82.7% ash content). Holothuroids had the thickest body-wall tissues and contained the lowest levels of mineralization in their body-walls. Crinoids and ophiuroids had high levels of mineralization in their arms. Asteroid body-wall tissues varied the most in thickness and ash content (0.9–3.9 mm in thickness and 29.2–55.5% in ash content). Body-wall tissues of 19 species of echinoderms were tested for their feeding deterrent properties against the marine fishes Lagodon rhomboides (Linnaeus) and Cyprinodon variegatus (Lacepede), as well as the decapod crustacean Libinia emarginata (Leach). Equivalent sized pieces of fresh body-wall tissue of 16 species of echinoderms caused observable feeding deterrence responses in at least two of the three model predators. There was no significant correlation between body-wall thickness or percent ash and its palatability to any of the three model predators. Agar pellets containing ethanolic body-wall extracts of 12 of 18 echinoderm species caused observable feeding deterrence responses in the fish L. rhomboides. In similar experiments with the arrow crab Stenorhyncus seticornis (Herbst), using carrageenan fish-meal blocks as food models, no differences in consumption of control fish-meal and experimental body-wall extract blocks were detected. Our findings indicate that invertebrate and vertebrate predators may respond quite differently to echinoderm body-wall extracts.

Bioactivity of echinoderm ethanolic body-wall extracts: an assessment of marine bacterial attachment and macroinvertebrate larval settlement

The ethanolic body-wall extracts of 16 species of echinoderms from 16 genera were screened for their ability to affect the attachment of the marine bacteria Deleya marina (Baumann) and Alteromonas luteo-violacea (Gauthier). Body-wall extracts were tested at concentrations which mimic mean natural tissue concentration, 3.0 mg/ml seawater, and four half-log dilutions of this initial concentration. The extracts of three echinoderm species caused significant inhibition of bacterial attachment, while extracts of eight species caused significant enhancement of attachment. The body-wall extract of the asteroid Goniaster tesselatus (Lamarck) displayed the most potent antimicrobial activity, completely inhibiting attachment of both bacterial species at a concentration of 3.0 mg/ml seawater. The ethanolic extracts of 20 echinoderm species were also tested at a similar range of concentrations for their ability to affect the settlement of cyprid larvae of the barnacle Balanus amphitrite (Darwin), and coronate larvae of the bryozoan Bugula neritina (Linné). All echinoderm extracts inhibited settlement of both barnacle and bryozoan larvae at the highest concentration tested (3.0 mg/ml seawater). Eleven of the 20 echinoderms tested (13 asteroids, 3 holothuroids, 3 ophiuroids and a crinoid) had body-wall extracts that inhibited settlement of competent barnacle and bryozoan larvae at concentrations as low as 0.12 mg/ml seawater. These extracted echinoderm compounds may function as non-toxic or toxic antifoulants, and to promote bacterial surface colonization, which could be valuable to the organisms disease resistance.

Actions of noradrenaline and some other biogenic amines on the body-wall muscles of an echiuroid, urechis unicinctus

1. Noradrenaline inhibited contractions of the body-wall strips of U. unicinctus in response to electrical pulse of stimulation and l-proline, while serotinin enhanced them. In addition, serotonin increased the rate of relaxation of twitch contraction. Octopamine enhanced the contractions but was less potent than serotonin. 2. Acetylcholine-contraction of the isolated inner circular body-wall muscle was also inhibited by noradrenaline and enhanced by serotonin. 3. Phentolamine, an α-adrenoreceptor blocker, potentiated tetanic contraction of the strip, though it did not alter twitch contraction. The inhibitory action of noradrenaline was blocked by phentolamine. 4. Noradrenaline hyperpolarized the fibre membrane of the inner circular muscle, while serotonin did not alter the membrane potential. The action of noradrenaline was blocked by phentolamine. 5. Bioassay of the body-wall extract and the ventral nerve-cord extract suggests that these extracts contain serotonin-like and noradrenaline-like substances, respectively. 6. These results suggest that noradrenaline and serotonin may be neurotransmitters or neurohormones modulating muscle contraction in the body wall of U. unicinctus.

Hydrolysis of juvenile hormone in diapausing and non-diapausing larvae of the southwestern corn borer,Diatraea grandiosella

The role of juvenile hormone (JH) esterases in relation to the diapause state of the southwestern corn borer,Diatraea grandiosella, was examined. The facultative larval diapause of this insect is dependent upon the presence of JH. Plasma, fat body, midgut, and body wall extracts metabolized [3H]JH I and [3H]JH III to JH-acid in vitro. JH-diol, JH-acid-diol, or conjugated polar metabolites were not detected. A longer half life of [3H]JH I was found in vitro in the plasma of diapausing larvae than in that of non-diapausing larvae. Although JH hydrolytic activity was relatively low in the plasma of pre-diapausing and diapausing larvae, systematic changes were observed suggesting that JH esterases may be involved in regulating the JH titer during this period. The JH hydrolytic activity found in the plasma of diapausing larvae was 3 to 5 times lower than that found in the plasma of mid-last instar non-diapausing larvae. Gel filtration profiles obtained from the plasma of diapausing and non-diapausing larvae suggested that JH esterases and α-naphthyl-acetate esterases are different enzymes. Multiple overlapping peaks of JH hydrolytic activity with an apparent molecular weight range of 43,000 to 75,000 were detected, whereas 2 separate peaks of α-naphthyl-acetate hydrolytic activity (apparent mol. wt. ca. 54,000, and 120,000) were detected. Gel filtration of supernatants of fat body indicated that JH was hydrolyzed at a lower rate by the fat body of pre-diapausing larvae than by that of non-diapausing larvae.

Proteins of the brain and body wall in larvae ofDrosophila melanogaster

Proteins of the brain and body wall cells of third instar larvae ofDrosophila melano-gaster have been examined by two-dimensional gel electrophoresis. Out of over 600 [35 S ]-labelled peptide spots seen in brain or body wall extracts, 517 were common to both; 61 spots were unique to brain and 66 unique to muscle. Glycoproteins were identified by soaking the gels in radioactive iodinated Concanavalin-A. Forty four Con-A binding glycoproteins were identifiable in the brain and 41 in the muscle extracts. Out of these, 8 glycoproteins of the brain and 8 of muscles appear to be tissue-specific.

The presence of alkenyl and alkyl glycerides, and choline and ethanolamine plasmalogens in a marine worm

1. 1. Crude lipid extracts of coelomic fluid, body wall and gut from the marine polychaete Nereis virens Sars were chromatographed on silica gel glass fibre sheets. 2. 2. Glyceryl ethers and neutral plasmalogens were found in unusually large amounts in coelomic fluid and body wall extracts but not in gut extracts. 3. 3. Phosphatidyl ethanolamine and phosphatidyl choline in all tissues gave a positive reaction to the plasmalogen test (method of Hack).

Trypsin and chymotrypsin inhibitors from Ascaris suum

Body wall and perienteric fluid of Ascaris suum were extracted with buffer, fractionated, and purified on carboxymethylcellulose. A trypsin inhibitor was eluted between pH 4.5 and 4.8 from perienteric fluid and body wall extract. The latter inhibited 3380 and 5170 μg of trypsin per milligram of protein, respectively. Two distinct chymotrypsin inhibitors were found. One inhibitor was eluted from carboxymethylcellulose between pH 4.3 and 4.6, while the second was eluted between pH 6.3 and 9.0. These inhibited 1930 and 2740 μg of chymotrypsin per milligram of protein, respectively.