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1.
PLoS One ; 17(9): e0274245, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36084062

RESUMO

The sea cucumber Holothuria (Metriatyla) scabra, known as sandfish, is a high-value tropical echinoderm central to the global bêche-de-mer (BDM) trade. This species has been heavily exploited across its natural range, with overharvesting and ineffective fishery management leaving stocks in the Pacific region heavily depleted. In Fiji, sandfish stocks have not recovered since a 1988 harvest ban, with surveys reporting declining populations and recruitment failure. Therefore, to inform fishery management policy for the wild sandfish resource and to guide hatchery-based restocking efforts, a high-resolution genomic audit of Fijian populations was carried out. A total of 6,896 selectively-neutral and 186 putatively-adaptive genome-wide SNPs (DArTseq) together with an independent oceanographic particle dispersal model were used to investigate genetic structure, diversity, signatures of selection, relatedness and connectivity in six wild populations. Three genetically distinct populations were identified with shallow but significant differentiation (average Fst = 0.034, p≤0.05), comprising (1) Lakeba island (Lau archipelago), (2) Macuata (Vanua Levu), and (3) individuals from Yasawa, Ra, Serua island and Kadavu comprising the final unit. Small reductions in allelic diversity were observed in marginal populations in eastern Fiji (overall mean A = 1.956 vs. Lau, A = 1.912 and Macuata, A = 1.939). Signatures of putative local adaptation were also discovered in individuals from Lakeba island, suggesting that they be managed as a discrete unit. An isolation-by-distance model of genetic structure for Fijian sandfish is apparent, with population fragmentation occurring towards the east. Hatchery-based production of juveniles is promising for stock replenishment, however great care is required during broodstock source population selection and juvenile releases into source areas only. The successful use of genomic data here has the potential to be applied to other sea cucumber species in Fiji, and other regions involved in the global BDM trade. While preliminary insights into the genetic structure and connectivity of sandfish in Fiji have been obtained, further local, regional and distribution-wide investigations are required to better inform conservation efforts, wild stock management and hatchery-based restocking interventions for this valuable invertebrate.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Equinodermos , Pesqueiros , Holothuria/genética , Humanos , Metagenômica , Pepinos-do-Mar/genética
2.
Int J Mol Sci ; 23(16)2022 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-36012278

RESUMO

Pollution in the seas and oceans is a global problem, which highlights emerging pollutants and plastics, specifically microplastics (MPs), which are tiny (1 µm to 5 mm) ubiquitous plastic particles present in marine environments that can be ingested by a wide range of organisms. Holothurians are benthic organisms that feed on sediment; therefore, they can be exposed to contaminants present in the particles they ingest. The objective was to evaluate the effects of human activity on Holothuria tubulosa through the study of biomarkers. Specimens were collected in three different areas throughout the island of Eivissa, Spain: (1) a highly urbanized area, with tourist uses and a marina; (2) an urbanized area close to the mouth of a torrent; (3) an area devoid of human activity and considered clean. The results showed a higher presence of microplastics (MPs) in the sediments from the highly urbanized area in relation to the other two areas studied. Similarly, a higher number of MPs were observed in the digestive tract of H. tubulosa from the most affected area, decreasing with the degree of anthropic influence. Both in the sediment and in the holothurians, fibers predominated with more than 75% of the items. In the three areas, mesoplastics were analyzed by means of FTIR, showing that the main polymer was polypropylene (27%) followed by low-density polyethylene (17%) and polystyrene (16%). Regarding the biomarkers of oxidative stress, the intestine of H. tubulosa from the most impacted areas showed higher catalase, superoxide dismutase (SOD), glutathione reductase (GRd), and glutathione S-transferase (GST) activities and reduced glutathione (GSH) levels compared to the control area. The intermediate area only presented significant differences in GRd and GST with respect to the clean area. The activities of acetylcholinesterase and the levels and malondialdehyde presented similar values in all areas. In conclusion, human activity evaluated with the presence of MPs induced an antioxidant response in H. tubulosa, although without evidence of oxidative damage or neurotoxicity. H. tubulosa, due to its benthic animal characteristics and easy handling, can be a useful species for monitoring purposes.


Assuntos
Holothuria , Poluentes Químicos da Água , Acetilcolinesterase , Animais , Biomarcadores/metabolismo , Monitoramento Ambiental , Glutationa Redutase , Atividades Humanas , Humanos , Intestinos/química , Microplásticos/toxicidade , Estresse Oxidativo , Plásticos , Poluentes Químicos da Água/análise
3.
Front Immunol ; 13: 927880, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35911686

RESUMO

Heavy-metal pollution has increasingly jeopardized the habitats of marine organisms including the sea cucumber, a seafloor scavenger vital to seawater bio-decontamination, ocean de-acidification and coral-reef protection. Normal physiology including immune functions of sea cucumbers is toxicologically modulated by marine metal pollutants such as cadmium (Cd). The processes underpinning Cd's toxic effects on immune systems in the sea cucumber, Holothuria leucospilota, are still poorly understood. To this end, we cloned and characterized a full-length caspase-9 (Hl-CASP9) cDNA in the sea cucumber, Holothuria leucospilota. Hl-CASP9 mRNA levels evolved dynamically during embryonic development. Coelomocytes, a type of phagocytic immune effectors central to H. leucospilota immunity, were found to express Hl-CASP9 mRNA most abundantly. Hl-CASP9 protein structurally resembles caspases-2 and -9 in both invertebrate and vertebrate species, comprising a CARD domain and a CASc domain. Remarkably, Hl-CASP9 was transcriptionally sensitive to abiotic oxidative stress inducers including hydrogen peroxide (H2O2), nitric oxide (•NO) and cadmium (Cd), but insensitive to immunostimulants including lipopolysaccharide (LPS), and poly(I:C). Overexpression of Hl-CASP9 augmented mitochondria-dependent apoptosis in HEK293T cells, while knock-down of Hl-CASP9 blunted Cd-induced coelomocyte apoptosis in vivo. Overall, we illustrate that an evolutionarily ancient caspase-9-dependent pathway exists to sensitize coelomocytes to premature cell death precipitated by heavy metal pollutants, with important implications for negative modulation of organismal immune response in marine invertebrates.


Assuntos
Apoptose , Cádmio , Caspase 9 , Holothuria , Animais , Apoptose/genética , Cádmio/toxicidade , Caspase 9/metabolismo , Poluentes Ambientais , Células HEK293 , Holothuria/genética , Holothuria/metabolismo , Humanos , Peróxido de Hidrogênio , RNA Mensageiro/genética , Pepinos-do-Mar/genética , Pepinos-do-Mar/metabolismo
4.
Adv Mar Biol ; 91: 1-286, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35777924

RESUMO

Holothuria scabra is one of the most intensively studied holothuroids, or sea cucumbers (Echinodermata: Holothuroidea), having been discussed in the literature since the early 19th century. The species is important for several reasons: (1) it is widely distributed and historically abundant in several shallow soft-bottom habitats throughout the Indo-Pacific, (2) it has a high commercial value on the Asian markets, where it is mainly sold as a dried product (beche-de-mer) and (3) it is the only tropical holothuroid species that can currently be mass-produced in hatcheries. Over 20 years have elapsed since the last comprehensive review on H. scabra published in 2001. Research on H. scabra has continued to accumulate, fuelled by intense commercial exploitation, and further declines in wild stocks over the entire distribution range. This review compiles data from over 950 publications pertaining to the biology, ecology, physiology, biochemical composition, aquaculture, fishery, processing and trade of H. scabra, presenting the most complete synthesis to date, including scientific papers and material published by local institutions and/or in foreign languages. The main goal of this project was to summarize and critically discuss the abundant literature on this species, making it more readily accessible to all stakeholders aiming to conduct fundamental and applied research on H. scabra, or wishing to develop aquaculture, stock enhancement and management programs across its geographic range.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Equinodermos , Ecologia , Pesqueiros
5.
Mar Drugs ; 20(6)2022 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-35736183

RESUMO

Fucosylated chondroitin sulfates (FCSs) FCS-BA and FCS-HS, as well as fucan sulfates (FSs) FS-BA-AT and FS-HS-AT were isolated from the sea cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera, respectively. Purification of the polysaccharides was carried out by anion-exchange chromatography on DEAE-Sephacel column. Structural characterization of polysaccharides was performed in terms of monosaccharide and sulfate content, as well as using a series of non-destructive NMR spectroscopic methods. Both FCSs were shown to contain a chondroitin core [→3)-ß-d-GalNAc-(1→4)-ß-d-GlcA-(1→]n bearing sulfated fucosyl branches at O-3 of every GlcA residue in the chain. These fucosyl residues were different in pattern of sulfation: FCS-BA contained Fuc2S4S, Fuc3S4S and Fuc4S at a ratio of 1:8:2, while FCS-HS contained these residues at a ratio of 2:2:1. Polysaccharides differed also in content of GalNAc4S6S and GalNAc4S units, the ratios being 14:1 for FCS-BA and 4:1 for FCS-HS. Both FCSs demonstrated significant anticoagulant activity in clotting time assay and potentiated inhibition of thrombin, but not of factor Xa. FS-BA-AT was shown to be a regular linear polymer of 4-linked α-L-fucopyranose 3-sulfate, the structure being confirmed by NMR spectra of desulfated polysaccharide. In spite of considerable sulfate content, FS-BA-AT was practically devoid of anticoagulant activity. FS-HS-AT cannot be purified completely from contamination of some FCS. Its structure was tentatively represented as a mixture of chains identical with FS-BA-AT and other chains built up of randomly sulfated alternating 4- and 3-linked α-L-fucopyranose residues.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Anticoagulantes/química , Anticoagulantes/farmacologia , Sulfatos de Condroitina/química , Fucose/química , Holothuria/química , Polissacarídeos/farmacologia , Pepinos-do-Mar/química , Sulfatos/farmacologia
6.
Int J Mol Sci ; 23(9)2022 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-35562879

RESUMO

Fucan sulfate I (FSI) from the sea cucumber Holothuria fuscopunctata was purified and its structure was clarified based on a bottom-up strategy. The unambiguous structures of a series of oligosaccharides including disaccharides, trisaccharides, and tetrasaccharides, which were released from mild acid hydrolysis of FSI, were identified by one-dimensional (1D)/two-dimensional (2D) nuclear magnetic resonance (NMR) and mass spectrometry (MS) analysis. All the glycosidic bonds in these oligosaccharides were presented as α1,3 linkages confirmed by correlated signals from their 1H-1H ROESY and 1H-13C HMBC spectra. The structural sequence of these oligosaccharides formed by Fuc2S4S, Fuc2S, and non-sulfated ones (Fuc0S), along with the general structural information of FSI, indicated that the structure of FSI could be elucidated as: [-L-Fuc2S4S-α1,3-L-Fuc(2S)-α1,3-L-Fuc2S-α1,3-L-Fuc0S-α1,3-1-]n. Moreover, the L-Fuc0S-α1,3-L-Fuc2S4S linkage in FSI was susceptible to be cleaved by mild acid hydrolysis. The antioxidant activity assays in vitro showed that FSI and the depolymerized product (dFSI') had potent activities for superoxide radical scavenging activity with IC50 of 65.71 and 83.72 µg/mL, respectively, while there was no scavenging effect on DPPH, hydroxyl and ABTS radicals.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Anticoagulantes/química , Antioxidantes/farmacologia , Holothuria/química , Oligossacarídeos/química , Polissacarídeos , Pepinos-do-Mar/química
7.
Carbohydr Polym ; 287: 119362, 2022 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-35422306

RESUMO

Fucosylated chondroitin sulfate was obtained from the sea cucumber Holothuria fuscogilva (FCShf). The structure was elucidated by NMR and HILIC-FTMS analysis. FCShf contained a chondroitin core chain [→3)-ß-D-GalNAc-(1 â†’ 4)-ß-D-GlcA-(1→]n, where the sulfation positions were the O-4 or O-6 of the GalNAc residues. The ratio of sulfated and non-sulfated GalNAc at O-6 was 1:2, while the ratio of GalNAc at O-4 was 1:1. 2,4-disulfated-fucose (Fuc2,4S), 4-sulfated-fucose (Fuc4S) and 3,4-disulfated-fucose (Fuc3,4S) were attached to the O-3 of GlcA with a molar ratio of 1.00: 0.62: 1.32. The FCShf could significantly promote the proliferative rate, NO production and neutral red uptake of RAW 264.7 cells within the concentration range of 10-300 µg/mL. Compared with the fucosylation and deacetylation degrees, the molecular weight of FCShf had markedly influence on the activation of RAW 264.7 cells. A decrease in molecular weight dramatically improved the immunoregulatory activities. Furthermore, FCShf activated RAW 264.7 cells through TLR-2/4-NF-κB pathway.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Sulfatos de Condroitina/química , Fucose/química , Holothuria/metabolismo , Camundongos , Células RAW 264.7 , Pepinos-do-Mar/química , Sulfatos/metabolismo
8.
Int J Mol Sci ; 23(6)2022 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-35328428

RESUMO

Apoptosis, also known as programmed cell death, is a biological process that is critical for embryonic development, organic differentiation, and tissue homeostasis of organisms. As an essential mitochondrial flavoprotein, the apoptosis-inducing factor (AIF) can directly mediate the caspase-independent mitochondrial apoptotic pathway. In this study, we identified and characterized a novel AIF-2 (HlAIF-2) from the tropical sea cucumber Holothuria leucospilota. HlAIF-2 contains a conserved Pyr_redox_2 domain and a putative C-terminal nuclear localization sequence (NLS) but lacks an N-terminal mitochondrial localization sequence (MLS). In addition, both NADH- and FAD-binding domains for oxidoreductase function are conserved in HlAIF-2. HlAIF-2 mRNA was ubiquitously detected in all tissues and increased significantly during larval development. The transcript expression of HlAIF-2 was significantly upregulated after treatment with CdCl2, but not the pathogen-associated molecular patterns (PAMPs) in primary coelomocytes. In HEK293T cells, HlAIF-2 protein was located in the cytoplasm and nucleus, and tended to transfer into the nucleus by CdCl2 incubation. Moreover, there was an overexpression of HlAIF-2-induced apoptosis in HEK293T cells. As a whole, this study provides the first evidence for heavy metal-induced apoptosis mediated by AIF-2 in sea cucumbers, and it may contribute to increasing the basic knowledge of the caspase-independent apoptotic pathway in ancient echinoderm species.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Apoptose , Fator de Indução de Apoptose/genética , Caspases , Células HEK293 , Humanos , Pepinos-do-Mar/genética
9.
Nutrients ; 14(4)2022 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-35215436

RESUMO

Cancer remains the primary cause of death worldwide. To develop less toxic anti-cancer drugs to relieve the suffering and improve the survival of cancer patients is the major focus in the anti-cancer field. To this end, marine creatures are being extensively studied for their anti-cancer effects, since extracts from at least 10% of the marine organisms have been shown to possess anti-tumor activities. As a classic Chinese traditional medicine, sea cucumbers and compounds extracted from the sea cucumbers, such as polysaccharides and saponins, have recently been shown to exhibit anti-cancer, anti-inflammatory, and anti-oxidant effects. Holothuria leucospilota (H. leucospilota) is a tropical edible sea cucumber species that has been successfully cultivated and farmed in large scales, providing a readily available source of raw materials to support the development of novel marine anti-cancer drugs. However, very few studies have so far been performed on the biological activities of H. leucospilota. In this study, we first investigated the anti-cancer effect of H. leucospilota protein on three cancer cell lines (i.e., HepG2, A549, Panc02) and three normal cell lines (NIH-3T3, HaCaT, 16HBE). Our data showed that H. leucospilota protein decreased the cell viabilities of HepG2, A549, HaCaT, 16HBE in a concentration-dependent manner, while Panc02 and NIH-3T3 in a time- and concentration-dependent manner. We also found that the inhibitory effect of H. leucospilota protein (≥10 µg/mL) on cell viability is near or even superior to EPI, a clinical chemotherapeutic agent. In addition, our data also demonstrated that H. leucospilota protein significantly affected the cell cycle and induced apoptosis in the three cancer cell lines investigated; in comparison, it showed no effects on the normal cell lines (i.e., NIH-3T3, HaCaT and 16HBE). Finally, our results also showed that H. leucospilota protein exhibited the excellent performance in inhibiting cell immigrations. In conclusion, H. leucospilota protein targeted the cancer cell cycles and induced cancer cell apoptosis; its superiority to inhibit cancer cell migration compared with EPI, shows the potential as a promising anti-cancer drug.


Assuntos
Antineoplásicos , Holothuria , Neoplasias , Saponinas , Pepinos-do-Mar , Animais , Antineoplásicos/farmacologia , Antioxidantes , Humanos , Neoplasias/tratamento farmacológico
10.
Molecules ; 27(2)2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-35056852

RESUMO

Saponins are plant and marine animal specific metabolites that are commonly considered as molecular vectors for chemical defenses against unicellular and pluricellular organisms. Their toxicity is attributed to their membranolytic properties. Modifying the molecular structures of saponins by quantitative and selective chemical reactions is increasingly considered to tune the biological properties of these molecules (i) to prepare congeners with specific activities for biomedical applications and (ii) to afford experimental data related to their structure-activity relationship. In the present study, we focused on the sulfated saponins contained in the viscera of Holothuria scabra, a sea cucumber present in the Indian Ocean and abundantly consumed on the Asian food market. Using mass spectrometry, we first qualitatively and quantitatively assessed the saponin content within the viscera of H. scabra. We detected 26 sulfated saponins presenting 5 different elemental compositions. Microwave activation under alkaline conditions in aqueous solutions was developed and optimized to quantitatively and specifically induce the desulfation of the natural saponins, by a specific loss of H2SO4. By comparing the hemolytic activities of the natural and desulfated extracts, we clearly identified the sulfate function as highly responsible for the saponin toxicity.


Assuntos
Holothuria/química , Saponinas/química , Saponinas/farmacologia , Sulfatos/química , Sulfatos/farmacologia , Vísceras/química , Álcalis/química , Animais , Bovinos , Cromatografia Líquida , Hemólise/efeitos dos fármacos , Hemolíticos/análise , Hemolíticos/química , Hemolíticos/isolamento & purificação , Hemolíticos/farmacologia , Hidrólise , Oceano Índico , Micro-Ondas , Saponinas/análise , Saponinas/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Relação Estrutura-Atividade , Sulfatos/análise , Sulfatos/isolamento & purificação , Espectrometria de Massas em Tandem
11.
Infect Disord Drug Targets ; 22(3): e170122200295, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35078399

RESUMO

BACKGROUND: An impressive treatment for toxoplasmosis is the combinatory use of sulfadiazine and pyrimethamine. However, both the drugs involve significant side effects and toxicity for the host. Therefore, the discovery of new anti-toxoplasma medications with high efficacy and less to no side effects is urgently needed. OBJECTIVE: This study aimed to evaluate the anti-toxoplasmic effects of Holothuria leucospilota (H. leucospilota) extract and TiO2NPs on the cell death of Toxoplasma gondii (T. gondii) tachyzoites in vitro and serum liver enzymes (AST, ALT, and ALP), and also to evaluate the immune response and production of IL-5, IFN-γ, and TNF-α in a mouse model. MATERIALS AND METHODS: The cytotoxicity of TiO2NPs and H. leucospilota extract against the tachyzoite of T. gondii was evaluated by the methyl thiazolyl tetrazolium (MTT) assay. The levels of serum TNF-α, IFN-γ, IL-5, and liver enzymes were measured, as well. All the groups were subjected to T. gondii, and the survival rate of experimental mice was evaluated. RESULTS: Our findings suggested in vivo and in vitro anti-toxoplasmic activity of TiO2NPs and H. leucospilota extract by inhibiting the proliferation and invasion of T. gondii tachyzoite. In addition, a significant increase in IFN-γ and TNF-α production was observed in mice treated with high doses of TiO2NPs and H. leucospilota extract. However, IL-5 levels decreased in TiO2NPs and H. leucospilota extract-treated mice. Our results also showed a highly significant increase (P < 0.05) in the levels of ALT, AST, and ALP in the groups injected with TiO2NPs and H. leucospilota extract, but not the control group. CONCLUSION: TiO2NPs and H. leucospilota extract have greater anti-toxoplasma effects in vitro and in vivo. These two compounds could be considered as a candidate for use against toxoplasmosis, both therapeutically and prophylactically.


Assuntos
Holothuria , Toxoplasma , Toxoplasmose , Animais , Humanos , Interleucina-5/farmacologia , Interleucina-5/uso terapêutico , Camundongos , Baço , Toxoplasmose/tratamento farmacológico , Fator de Necrose Tumoral alfa/farmacologia , Fator de Necrose Tumoral alfa/uso terapêutico
12.
Mar Pollut Bull ; 175: 113134, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34823866

RESUMO

This study investigated the abundances and characteristics of microplastics in sediments and sandfish (Holothuria scabra) in Lampung and Sumbawa, Indonesia. Microplastics were found in 89.02% of all sandfish samples, with an average abundance of 2.01 ± 1.59 particles individual-1. The abundance of microplastics was 58.42 ± 24.33 particles kg-1 in surface sediments. Furthermore, there was a positive relationship between the abundance of microplastics in sandfish and sediments. Fragments and fibers with small-sized microplastics (300-1000 µm) were the most abundant types found in sandfish and sediments. Fourier-transform infrared (FTIR) analysis showed that polyethylene (30.08%), polypropylene (30.08%), polyurethane (12.20%), and polyethylene terephthalate (8.94%) were the most abundant polymers in the samples. Our results strongly indicate that microplastics in Lampung and Sumbawa originate from the fragmentation of large plastics. Better solid waste management in Indonesia is needed to reduce plastic waste leakage, which could become microplastics.


Assuntos
Holothuria , Poluentes Químicos da Água , Animais , Ingestão de Alimentos , Monitoramento Ambiental , Sedimentos Geológicos , Indonésia , Microplásticos , Plásticos/análise , Poluentes Químicos da Água/análise
13.
J Exp Zool B Mol Dev Evol ; 338(3): 155-169, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-34813182

RESUMO

The respiratory trees present only in the class Holothuroidea and the rest of the echinoderms lack it. Only some holothurian species have the ability to regenerate their respiratory trees after autotomy. Therefore, respiratory trees could be considered as a suitable model to assess the regeneration mechanisms in animals. In the present study, the respiratory tree regeneration after posterior evisceration were examined in Holothuria parva during 75 days. Since autotomy reduces antioxidant defense in the organisms, in the present study alterations of antioxidant enzymes were also evaluated during the experiment. H. parva is the dominant intertidal species distributed in the north of the Persian Gulf. In the present study, H. parva ejected the left respiratory tree, the digestive tract and supportive mesenteries from the anus, about 1-2 min after potassium chloride injection. The closure of the opening at the posterior ends of the body was the first reaction to the injury. Seven days after evisceration, the small bud formed on the dorsal side of the cloaca which was covered with the coelomic epithelium of cloaca. The coelomic epithelium started to proliferate to undifferentiated cells on the apex of the buds. The primary respiratory tree consisted of the luminal cuboidal epithelium and thin connective tissue surrounded by the slender coelomic epithelium. This preliminary organ was observed at the apex of the buds, 13 days after evisceration. Gradually, myoepithelial cells arranged around a longitudinal axis and formed a circular muscle. The primitive branches of primary respiratory tree started to form 18 days after evisceration. Forty days after evisceration, the luminal epithelium of the respiratory tree had the same appearance as the intact luminal epithelium. The regenerated respiratory tree was histomorphologically very similar to an intact respiratory tree 56 days postevisceration, but unlike that, it was not yet wrapped around the intestine and was completely separate from it. Despite the development of the regenerating respiratory tree, no wrapping around the intestine was observed until the end of the experiment. According to the results, the activity of the catalase (CAT) and superoxide dismutase (SOD) in the muscle homogenate was significantly higher than the control 5 days after evisceration. The CAT and SOD levels gradually decreased in eviscerated animals. The lipid peroxidation level followed a decreasing trend in the eviscerated animals during the experiment. However, its value reduced to the control level at the end of the experiment.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Antioxidantes , Holothuria/fisiologia , Superóxido Dismutase , Árvores
14.
Nutr Cancer ; 74(4): 1511-1518, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34459328

RESUMO

The incidence of cancer has exhibited an increasing trend in recent years because of many reasons such as environmental and nutritional factors. There is a great need for the development of new and natural molecules with lower side effects in the therapy of cancer. It was aimed to evaluate the antiproliferative effect of semi-purified triterpene glycosides of Holothuria poli on different human cancer cell lines. The body walls of H. poli as the main sources of saponins were used and the saponin content of the extract was characterized by MALDI-TOF/MS. The antiproliferation activity of the characterized extract was tested on cancer cell lines. The extract showed antiproliferative effect on the studied cancer cell lines. The mass analysis results reveal that Holothurin A is one of the saponins within the extract. The measured IC50 values were found as 31.41 ± 2.20, 77.45 ± 0.23, and 34.79 ± 0.90 µg mL-1 for HT-29, UPCI-SCC-131, and T84 cell lines, respectively. H. poli secretes not only specific saponins but also a cocktail of them. Specific versus. cocktails of the saponins and by also applying organic modification must be studied in further research to understand their mechanisms in the antiproliferation studies since this paper reveals promising results.


Assuntos
Holothuria , Saponinas , Triterpenos , Animais , Linhagem Celular Tumoral , Holothuria/metabolismo , Humanos , Estrutura Molecular , Extratos Vegetais/metabolismo , Saponinas/farmacologia , Triterpenos/farmacologia
15.
Carbohydr Polym ; 275: 118694, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34742421

RESUMO

The elucidation of precise structure of fucoidan is essential for understanding their structure-function relationship and promoting the development of marine drugs. In this work, we firstly reported the oligosaccharide mapping of fucoidan from Holothuria floridana using a combination of hydrothermal depolymerization, hydrophilic interaction liquid chromatography (HILIC) coupled with electrospray mass spectrometry (ESI-FTMS) and high energy collision-induced dissociation (HCD-MS/MS) and 2D NMR analysis. With careful selection of fully deprotonated molecular ions of fucoidan oligosaccharides and their NaBD4 reduced alditols, HILIC-ESI-HCD-MS/MS provided structurally relevant glycosidic product ions with no sulfate loss for definitive assignment of sequence and sulfation pattern of all the oligosaccharides and their isomers from dp2 to dp7 from hydrothermal depolymerization. The oligosaccharide mapping clarified the structure of fucoidan with various oligosaccharide domains with 2,4-di-O-sulfated and 2-O sulfated fucose residues.


Assuntos
Oligossacarídeos/química , Polissacarídeos/química , Pepinos-do-Mar/química , Animais , Cromatografia Líquida/métodos , Holothuria/química , Interações Hidrofóbicas e Hidrofílicas , Isomerismo , Espectroscopia de Ressonância Magnética/métodos , Estrutura Molecular , Polimerização , Espectrometria de Massas por Ionização por Electrospray/métodos , Sulfatos/química , Espectrometria de Massas em Tandem/métodos
16.
J Neurosci Methods ; 364: 109358, 2021 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-34537226

RESUMO

BACKGROUND: Among animal phyla, some of the least studied nervous systems are those of the phylum Echinodermata. Part of the problem lies in that most of their nervous components are embedded in the body wall that has calcareous skeletal components. NEW METHOD: We have developed a novel technique for the successful isolation of the radial nerve cords (RNCs) and an in vitro system where the isolated RNCs can be cultured and are amenable to experimental manipulation. Here we use this system to isolate the RNC of the sea cucumber Holothuria glaberrima as a way to extend our studies on its regeneration capabilities. RESULTS: The RNCs can be isolated from the surrounding tissues by collagenase treatment. The explants obtained following enzymatic dissociation can be kept in culture for up to 2 weeks. Histological and immunohistochemical studies show that the explants maintain a stable number of cells with little proliferation or apoptosis throughout the culture incubation period. The main change observed in RNCs in vitro is a progressive dedifferentiation of radial glia-like cells. This dedifferentiation corresponds to the first step in the regeneration response to injury that has been described in vivo. COMPARISON WITH EXISTING METHODS: There are no existing methods to isolate and culture echinoderm radial nerve cord. CONCLUSIONS: The described protocol provides a unique tool to obtain easily accessible RNC from holothurians to perform cellular, biochemical, and genomic experiments in the echinoderm nervous system without interference of adjacent tissues. The technique provides a unique opportunity to study the dedifferentiation response associated with the regeneration of the nervous system in echinoderms.


Assuntos
Holothuria , Nervo Radial , Animais , Apoptose , Células Ependimogliais
17.
Artigo em Inglês | MEDLINE | ID: mdl-34488170

RESUMO

The sandfish Holothuria scabra is a high-value tropical sea cucumber species representing a major mariculture prospect across the Indo-Pacific. Advancements in culture technology, rearing, and processing present options for augmenting capture production, stock restoration, and sustainable livelihood activities from hatchery-produced sandfish. Further improvements in mariculture production may be gained from the application of genomic technologies to improve performance traits such as growth. In this study, we performed de novo transcriptome assembly and characterization of fast- and slow-growing juvenile H. scabra from three Philippine populations. Analyses revealed 66 unigenes that were consistently differentially regulated in fast-growing sandfish and found to be associated with immune response and metabolism. Further, we identified microsatellite and single nucleotide polymorphism markers potentially associated with fast growth. These findings provide insight on potential genomic determinants underlying growth regulation in early juvenile sandfish which will be useful for further functional studies.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Perfilação da Expressão Gênica , Holothuria/genética , Repetições de Microssatélites/genética , Pepinos-do-Mar/genética , Transcriptoma
18.
Dev Biol ; 480: 105-113, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34481794

RESUMO

Regeneration is a key developmental process by which organisms recover vital tissue and organ components following injury or disease. A growing interest is focused on the elucidation and characterization of the molecular mechanisms involved in these regenerative processes. We have now analyzed the possible role of the Wnt/ß-catenin pathway on the regeneration of the intestine in the sea cucumber Holothuria glaberrima. For this we have studied the expression in vivo of Wnt-associated genes and have implemented the use of Dicer-substrate interference RNA (DsiRNA) to knockdown the expression of ß-catenin transcript on gut rudiment explants. Neither cell dedifferentiation nor apoptosis were affected by the reduction of ß-catenin transcripts in the gut rudiment explants. Yet, the number of proliferating cells decreased significantly following the interference, suggesting that the Wnt/ß-catenin signaling pathway plays a significant role in cell proliferation, but not in cell dedifferentiation nor apoptosis during the regeneration of the intestine. The development of the in vitro RNAi protocol is a significant step in analyzing specific gene functions involved in echinoderm regeneration.


Assuntos
Intestinos/metabolismo , Músculo Esquelético/fisiologia , Via de Sinalização Wnt/fisiologia , Animais , Apoptose/fisiologia , Desdiferenciação Celular/fisiologia , Proliferação de Células/genética , Holothuria/metabolismo , Holothuria/fisiologia , Intestinos/crescimento & desenvolvimento , Músculo Esquelético/metabolismo , Regeneração/fisiologia , Pepinos-do-Mar/metabolismo , Pepinos-do-Mar/fisiologia , Proteínas Wnt/metabolismo , Via de Sinalização Wnt/genética , beta Catenina/metabolismo
20.
Artigo em Inglês | MEDLINE | ID: mdl-34303260

RESUMO

Ambient temperature is an important abiotic factor that influences growth performance and physiological functions in sea cucumbers. To understand the molecular responses of the sea cucumber Holothuria leucospilota to acute temperature stress, we performed a de novo transcriptome analysis of body wall tissue from H. leucospilota exposed to 2 hoursh of acute heat (35 ± 1 °C) and cold stress (15 ± 1 °C). A total of 99,015 unigenes were obtained after assembly of the sequenced reads. Compared with a control group maintained at 25.0 ± 1 °C, 1169 differentially expressed unigenes (DEGs) were identified after heat stress, 781 were up-regulated and 388 were down-regulated. After cold stress, 1464 DEGs were identified; 900 were up-regulated and 564 were down-regulated. The annotation of DEGs revealed that heat shock proteins play important roles in protecting H. leucospilota from high temperature stress. Furthermore, KEGG pathway enrichment analysis showed that the categories: "Ribosome" and "Protein processing in endoplasmic reticulum" were strongly affected by heat stress. These two pathways are associated with biosynthesis and processing of proteins, and refolding of misfolded proteins. The lipid metabolism pathways "Sphingolipid metabolism" and "Ether lipid metabolism", were affected by cold stress. The RNA-Seq results for eight selected DEGs were verified the expression by quantitative real-time PCR analysis. Our results will improve the understanding of the molecular response mechanisms of H. leucospilota to ambient temperature stress.


Assuntos
Holothuria , Pepinos-do-Mar , Animais , Perfilação da Expressão Gênica , Holothuria/genética , Pepinos-do-Mar/genética , Temperatura , Transcriptoma
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