Sphingosines Derived from Marine Sponge as Potential Multi-Target Drug Related to Disorders in Cancer Development.

Haliclona tubifera, marine sponge species abundant in Brazilian coastline, presents only a few papers published in the literature. Recently, we have reported the isolation of two modified C18 sphingoid bases: (2R,3R,6R,7Z)-2-aminooctadec-7-ene-1,3, 6-triol and and (2R,3R,6R)-2-aminooctadec-1,3,6-triol. In order to continue our research, in this work aimed at the biological investigation of fractions that led to the isolation of these compounds. We evaluated the cytotoxic effect of marine sponge H. tubifera fractions in glioma (U87) and neuroblastoma (SH-SY5Y) human cell lines. In addition, considering the link between cancer, imbalance of reactive oxygen species and coagulation disorders, we also investigated the in vitro effects on blood coagulation and their redox properties. We showed that the ethyl acetate (EtOAc) fraction, rich in sphingoid bases, had important cytotoxic effects in both cancer cell lines with an IC50 < 15 µg/mL and also can inhibit the production of peroxyl radicals. Interestingly, this fraction increased the recalcification time of human blood, showing anticoagulant properties. The present study indicates the sphingosines fraction as a promising source of chemical prototypes, especially multifunctional drugs in cancer therapy.

Cytotoxic effects on tumour cell lines of fatty acids from the marine sponge Scopalina ruetzleri.

OBJECTIVES: Marine sponges are among the most promising sources of chemically diversified fatty acids (FAs). In addition, several studies have shown the effect of polyunsaturated FAs on cancer therapy. This research carried out a biological and chemical evaluation of the sponge Scopalina ruetzleri collected on the South Brazilian coastline. METHODS: Bioassay-guided fractionation of S. ruetzleri was performed in human glioma (U87) and neuroblastoma (SH-SY5Y) cell lines, and the in-vitro effects on free radicals were evaluated. KEY FINDINGS: The ethyl acetate fraction of S. ruetzleri showed promising cytotoxic effects in cancer cell lines, with IC50 < 20 µg/ml. Fingerprint (1) H Nuclear Magnetic Resonance (NMR) analysis showed that this fraction is mainly constituted of FAs. Through FA methyl ester analysis, it was possible to identify 32 FAs. In addition, some minor unusual FAs for the marine biosphere were identified. The results of conjugated dienes method showed that FAs fraction, at concentrations above 50 µg/ml, has a pro-oxidant effect, indicating that lipid peroxidation may be partially responsible for the mechanism of cytotoxicity on cancer cells. CONCLUSION: This work also contributes to studies that focus on the application of FAs on cancer therapy as a new adjuvant to radio or chemotherapy, or as a chemotherapeutic agent.

Biological activities of ACL-I and physicochemical properties of ACL-II, lectins isolated from the marine sponge Axinella corrugata.

Lectin II from the marine sponge Axinella corrugata (ACL-II) was purified by affinity chromatography on rabbit erythrocytic stroma incorporated into a polyacrylamide gel, followed by gel filtration on Ultrogel AcA 44 column. Purified ACL-II is a lectin with an Mr of 80 kDa and 78 kDa, estimated by SDS-PAGE and by FPLC on Superose 12 HR column, respectively. ACL-II mainly agglutinates native rabbit erythrocytes and this hemagglutinating activity is independent of Ca(2+), Mg(2+) and Mn(2+), but is inhibited by d-galactose, chitin and N-acetyl derivatives, with the exception of GalNAc. ACL-II is stable for up to 65 °C for 30 min, with a better stability at a pH range of 2 to 6. In contrast, ACL-I displays a strong mitogenic and cytotoxic effect.

Current status on natural products with antitumor activity from Brazilian marine sponges.

Over the last few years, samples from the marine environment have been screened for a variety of compounds with different biological activities. Among all marine organisms, sponges represent one of the most promising sources of leads in the research of new cancer drugs. However, there are few reports on screening Brazilian marine sponges for biological activities. In the following review, the current status of natural product research relating to Brazilian marine sponges is summarized, particularly for compounds demonstrating potential antitumor activity.

Immunohistochemical localization of an N-acetyl amino-carbohydrate specific lectin (ACL-I) of the marine sponge Axinella corrugata.

The N-acetyl amino-carbohydrate specific lectin (ACL-I) was previously identified and purified by us from the marine sponge Axinella corrugata (phylum Porifera, class Demospongiae). The distribution of the specific lectin within the tissue of the sponge was studied by bright-field optical microscopy immunohistochemistry in order to better understand its physiological role in the sponge. Polyclonal antibodies were raised against purified ACL-I in mice and tested by Western blot technique. The immunohistochemical analysis of ACL-I in cross sections of A. corrugata showed that this lectin is found inside the denominated spherulous cells, which contain vesicles that store the lectin. Some evidence is shown that ACL-I might also be present in the extracellular matrix. It was not possible to demonstrate by the immunohistochemical technique if ACL-I is colocalized in both the plasma membrane and in the cytoplasm of the spherulous cells.

Brazilian marine sponge Polymastia janeirensis induces apoptotic cell death in human U138MG glioma cell line, but not in a normal cell culture.

Marine sponges have been prominently featured in the area of cancer research. Here, we examined the anti-proliferative effects of crude extracts (aqueous and organic) of the Brazilian marine sponge Polymastia janeirensis in the U138MG human glioma cell line. Moreover, we examined the effects of extracts on selective cytotoxicity in the glioma cells in comparison with a normal cell culture. Exposure of glioma cells to treatments (24 h) resulted in cell number decrease at all doses tested, with both aqueous and organic extracts (IC(50) <20 and <30 microg/ml, respectively). Parallel to this result, sponge extracts reduced glioma cell viability (IC(50) <15 microg/ml for both extracts). However, higher doses (50 and 100 microg/ml) induced a stronger cytotoxic effect when compared to the lower dose tested (10 microg/ml), inhibiting more than 80% of cellular growth and viability. Propidium iodide uptake and flow cytometry analysis further showed that sponge extracts caused necrosis in the glioma cell line at higher doses, while a high percentage of apoptotic glioma cells were observed at 10 microg/ml. Moreover, apoptosis was prevented by the pan-caspase inhibitor Z-VAD, suggesting that marine sponge extracts, at lower doses, induce caspase-dependent apoptosis in U138MG glioma cells. Surprisingly the extracts herein tested were more effective than temozolomide, a potent inductor of apoptosis used for the treatment of malignant gliomas. Furthermore, our results suggested a selectivity cytotoxic effect on glioma cell line in comparison with a normal cell culture, since the effect on viability found in glioma cells was not observed in astrocyte cultures with the lower dose (10 microg/ml). Thus, this marine sponge may be considered a good candidate for development of new cancer medicines with antitumor activity against gliomas.

Extracts of marine sponge Polymastia janeirensis induce oxidative cell death through a caspase-9 apoptotic pathway in human U138MG glioma cell line.

We have studied the apoptotic pathway activated in response to marine sponge extracts of Polymastia janeirensis. The effect on intracellular ROS production was also examined. Exposure of U138MG glioma cell line to doses higher than 5 microg/mL has decreased glioma cell viability, with an IC(50) <15 microg/mL for both aqueous and organic extracts. However, extracts at higher doses (50 and 100 microg/mL) have stronger cytotoxic effects, decreasing more than 90% of glioma cell viability. The antioxidant Trolox (100 microM) reversed the cell death percentage induced by extracts at 10 and 25 microg/mL. The type of cell death induced by such high doses was predominantly necrosis, while a high percentage of apoptotic glioma cells was found at 10 microg/mL. Moreover, inhibition of caspase-8 with Z-IETD (a caspase-8 inhibitor) had no effect on the amount of apoptosis induced by 10 microg/mL, but inhibition of caspase-9 with Z-LEHD (a caspase-9 inhibitor) decreased apoptosis. We also observed a dose-dependent increase in ROS production, and similarly to effects observed on viability of glioma cells, and on cell death, higher doses also had more severe effects. Co-treatment with Trolox significantly reduced ROS production by extracts at doses lower than 50 microg/mL. This is a first report demonstrating that marine sponge extracts of P. janeirensis induce oxidative cell death through a caspase-9 apoptotic pathway.

ACL-I, a lectin from the marine sponge Axinella corrugata: isolation, characterization and chemotactic activity.

The lectin from the marine sponge Axinella corrugata (ACL-I) was purified by affinity chromatography on rabbit erythrocytic stroma incorporated into a polyacrylamide gel followed by gel filtration on Ultrogel AcA 44 column. Purified ACL-I is a hexameric glycoprotein with a Mr of 82.3 kDa estimated by SDS-PAGE and 78.5 kDa by FPLC on Superose 12 HR column. The pI of lectin is 6.3 and ACL-I is constituted of 13.9 kDa similar subunits some of them linked by disulphide bridges. This lectin agglutinates native rabbit, goat and dog erythrocytes and in less extent human erythrocytes. The hemagglutinating activity is independent of Ca(2+), Mg(2+) and Mn(2+), but it is strongly inhibited by carbohydrates containing N-acetyl groups. ACL-I is stable up to 70 degrees C for 30 min, with optimum pH between 7 and 8, and it is also resistant to enzymatic proteolysis in vitro. In the presence of reducing or denaturant agents, the lectin activity decreases. ACL-I displays chemotactic effect on rat neutrophil in vitro which is inhibited by N-acetyl-d-glucosamine.

New records of the genus Agelas Duchassaing & Michelotti, 1864 (Porifera, Agelasida) off the Amazon River mouth, Brazil, Southwestern Atlantic / Novos registros do gênero Agelas Duchassaing & Michelotti, 1864 (Porifera, Agelasida) ao largo da desembocadura do Rio Amazonas, Brasil, Atlântico Sudoeste

This work provides new information on agelasid sponges found on the continental shelf off northern Brazil. Agelas sceptrum (Lamarck, 1815) and Agelas wiedenmayeri Alcolado, 1984 have their first record for the Brazilian coast. Agelas dispar Duchassaing & Michelotti, 1864 and Agelas schmidti Wilson, 1902, previously recorded from Brazil, are cited for the first time off the mouth of the Amazon River.

Este trabalho fornece novas informações sobre esponjas agelasidas encontradas na costa norte da plataforma continental brasileira. Agelas sceptrum (Lamarck, 1815) e Agelas wiedenmayeri Alcolado, 1984 têm seu primeiro registro para a costa brasileira. Agelas dispar Duchassaing & Michelotti, 1864 e Agelas schmidti Wilson, 1902, registradas anteriormente na costa brasileira, são citadas pela primeira vez ao largo da desembocadura do Rio Amazonas.

In vitro antiviral activity of marine sponges collected off Brazilian coast.

This paper describes the in vitro antiviral evaluation of 27 different marine sponges (Porifera) collected off Brazilian coastline in the search for novel drug leads. With these sponges aqueous and organic extracts were prepared and tested for anti-herpetic (HSV-1, KOS strain), anti-adenovirus (human AdV serotype 5) and anti-rotavirus (simian RV SA11) activities. The evaluation of the cytotoxicity and potential antiviral activity of these extracts were performed by using MTT assay. Results were expressed as 50% cytotoxicity (CC50) and 50% effective (EC50) concentrations, respectively, in order to calculate the selectivity indices (SI=CC50/EC50) of each extract. From the 40 sponge extracts tested, 17 extracts showed antiviral action in different degrees. The results concerning the antiviral activity were obtained by using three different strategies: (1) simultaneous assay, when sponge extracts were added to the cells at the same time of the viruses; (2) pre treatment assay, when sponge extracts were added to the cells 15 h prior to the viruses infection; and (3) post treatment assay, when the viruses were added to the cells and remained during 2 h prior to the addition of sponge extracts. The antiviral assays with HSV-1/KOS and AdV-5 showed more promising results when the pre treatment test was employed. In relation to the RV-SA11 virus, only the simultaneous assay showed antiviral activity. The extracts presenting the most promising results were the aqueous extracts of Cliona sp., Agelas sp.2, Tethya sp., Axinella aff corrugata, Polymastia janeirensis and Protosuberites sp., and these extracts deserve special attention in further studies.

Recruitment and colonization of colonial ascidians (Tunicata: Ascidiacea) on intertidal rocks in Northeastern Brazil

Recruitment and colonization of colonial ascidians, starting on natural scraped substratum, was observed throughout one year, in quadrats marked on sea- and continental-rock faces in the sublittoral fringe of an exposed beach in the Northeast of Brazil. The species of the Didemnidae, Didemnum duplicatum Monniot, 1983, D. psammathodes Sluiter, 1895, and Polysyncraton amethysteum Van Name, 1902, and an unidentified species of Polycitoridae, Eudistoma sp. 1, were recruited. Presenting continuous recruitment of all species, the density of living and dead recruits and colonies was similar on both faces of the rocks. The highest rainfall during winter, affected significantly the recruits density of D. duplicatum and D. psammathodes. Different permanence periods were observed for colonies of each species. The longest permanence periods were assigned to the colonies of P. amethysteum and D. psammathodes, extending up to four and five months, respectively.

A partir de substrato natural raspado, o recrutamento e a colonização de ascídias coloniais foi observado ao longo de um ano, em unidades amostrais delimitadas sobre as faces mar e continente de rochas situadas na franja do infralitoral de uma praia exposta no nordeste do Brasil. Foram recrutadas as espécies de Didemnidae, Didemnum duplicatum Monniot, 1983, D. psammathodes Sluiter, 1895 e Polysyncraton amethysteum Van Name, 1902 e uma espécie não-identificada de Polycitoridae, Eudistoma sp. 1. Apresentando recrutamento contínuo de todas as espécies, a densidade de colônias e de recrutas vivos e mortos foi similar nas duas faces das rochas. Durante o inverno, a densidade de recrutas de D. duplicatum e de D. psammathodes foi significativamente afetada pela intensa pluviosidade. As colônias de cada espécie apresentaram diferentes períodos de permanência sobre as rochas. Com duração de quatro e cinco meses, os maiores períodos de permanência foram assinalados para as colônias de P. amethysteum e D. psammathodes, respectivamente.

Spongia (Heterofibria) catarinensis sp. nov. (Porifera, Spongiidae) no litoral de Santa Catarina, Brasil

A new species, Spongia (Heterofibria) catarinensis, is described from Ilha das Aranhas (27º29'077''S, 48º21'380''W), State of Santa Catarina, Brazil. It differs from the other southwestern Atlantic species of this genus by external morphology and architecture. The new species is characterized for presenting massive form, incrusting with lobate projections; primary fibres measuring 60-100 µm; secondary fibres 11,5-69 µm; pseudotertiary fibres 2,3-23 µm and distance between primary fibres 391-920 µm; distance between secondary/pseudotertiary fibres 92-575 µm; secondary/pseudotertiary fibres constituting polygonal meshes 30-700 µm diameter. This species belongs to subgenus Heterofibria Cook & Bergquist, 2001 for displaying a clear dichotomy of its fibres.

Uma nova espécie, Spongia (Heterofibria) catarinensis, é descrita para a Ilha das Aranhas (27º29'077''S, 48º21'380''W), Estado de Santa Catarina, Brasil. Difere das outras espécies do gênero do Atlântico sudoeste pela morfologia externa e arquitetura. A nova espécie é caracterizada por apresentar forma massiva, incrustante com projeções lobulares; fibras primárias medindo 60-100 µm; fibras secundárias 11,5-69 µm; fibras pseudoterciárias 2,3-23 µm e distância entre fibras primárias 391-920 µm; distância entre fibras secundárias/pseudoterciárias 92-575 µm; fibras secundárias/pseudoterciárias constituindo malhas poligonais com 30-700 µm de diâmetro. A espécie pertence ao subgênero Heterofibria Cook & Bergquist, 2001 por apresentar uma clara dicotomia de suas fibras.

Spongia (Heterofibria) catarinensis sp. nov. (Porifera, Spongiidae) no litoral de Santa Catarina, Brasil

A new species, Spongia (Heterofibria) catarinensis, is described from Ilha das Aranhas (27º29'077''S, 48º21'380''W), State of Santa Catarina, Brazil. It differs from the other southwestern Atlantic species of this genus by external morphology and architecture. The new species is characterized for presenting massive form, incrusting with lobate projections; primary fibres measuring 60-100 µm; secondary fibres 11,5-69 µm; pseudotertiary fibres 2,3-23 µm and distance between primary fibres 391-920 µm; distance between secondary/pseudotertiary fibres 92-575 µm; secondary/pseudotertiary fibres constituting polygonal meshes 30-700 µm diameter. This species belongs to subgenus Heterofibria Cook & Bergquist, 2001 for displaying a clear dichotomy of its fibres.

Uma nova espécie, Spongia (Heterofibria) catarinensis, é descrita para a Ilha das Aranhas (27º29'077''S, 48º21'380''W), Estado de Santa Catarina, Brasil. Difere das outras espécies do gênero do Atlântico sudoeste pela morfologia externa e arquitetura. A nova espécie é caracterizada por apresentar forma massiva, incrustante com projeções lobulares; fibras primárias medindo 60-100 µm; fibras secundárias 11,5-69 µm; fibras pseudoterciárias 2,3-23 µm e distância entre fibras primárias 391-920 µm; distância entre fibras secundárias/pseudoterciárias 92-575 µm; fibras secundárias/pseudoterciárias constituindo malhas poligonais com 30-700 µm de diâmetro. A espécie pertence ao subgênero Heterofibria Cook & Bergquist, 2001 por apresentar uma clara dicotomia de suas fibras.

Recruitment and colonization of colonial ascidians (Tunicata: Ascidiacea) on intertidal rocks in Northeastern Brazil

Recruitment and colonization of colonial ascidians, starting on natural scraped substratum, was observed throughout one year, in quadrats marked on sea- and continental-rock faces in the sublittoral fringe of an exposed beach in the Northeast of Brazil. The species of the Didemnidae, Didemnum duplicatum Monniot, 1983, D. psammathodes Sluiter, 1895, and Polysyncraton amethysteum Van Name, 1902, and an unidentified species of Polycitoridae, Eudistoma sp. 1, were recruited. Presenting continuous recruitment of all species, the density of living and dead recruits and colonies was similar on both faces of the rocks. The highest rainfall during winter, affected significantly the recruits density of D. duplicatum and D. psammathodes. Different permanence periods were observed for colonies of each species. The longest permanence periods were assigned to the colonies of P. amethysteum and D. psammathodes, extending up to four and five months, respectively.

A partir de substrato natural raspado, o recrutamento e a colonização de ascídias coloniais foi observado ao longo de um ano, em unidades amostrais delimitadas sobre as faces mar e continente de rochas situadas na franja do infralitoral de uma praia exposta no nordeste do Brasil. Foram recrutadas as espécies de Didemnidae, Didemnum duplicatum Monniot, 1983, D. psammathodes Sluiter, 1895 e Polysyncraton amethysteum Van Name, 1902 e uma espécie não-identificada de Polycitoridae, Eudistoma sp. 1. Apresentando recrutamento contínuo de todas as espécies, a densidade de colônias e de recrutas vivos e mortos foi similar nas duas faces das rochas. Durante o inverno, a densidade de recrutas de D. duplicatum e de D. psammathodes foi significativamente afetada pela intensa pluviosidade. As colônias de cada espécie apresentaram diferentes períodos de permanência sobre as rochas. Com duração de quatro e cinco meses, os maiores períodos de permanência foram assinalados para as colônias de P. amethysteum e D. psammathodes, respectivamente.

Esponjas (Demospongiae, Halichondrida) da costa do Maranhão, Brasil

Three species of Halichondrida, Dragmacidon reticulatus (Ridley & Dendy, 1886) (Axinellidae), Myrmekioderma rea (Laubenfels, 1934) (Desmoxyidae) and Topsentia ophiraphidites (Laubenfels, 1934) (Halichondriidae), collected from 30 to 184 m depths, were recorded for the first time from State of Maranhão, north-northeast coast of Brazilian shelf.

Esponjas (Demospongiae, Halichondrida) da costa do Maranhão, Brasil

Three species of Halichondrida, Dragmacidon reticulatus (Ridley & Dendy, 1886) (Axinellidae), Myrmekioderma rea (Laubenfels, 1934) (Desmoxyidae) and Topsentia ophiraphidites (Laubenfels, 1934) (Halichondriidae), collected from 30 to 184 m depths, were recorded for the first time from State of Maranhão, north-northeast coast of Brazilian shelf.

Esponjas (Demospongiae, Halichondrida) da costa do Maranhão, Brasil

Three species of Halichondrida, Dragmacidon reticulatus (Ridley & Dendy, 1886) (Axinellidae), Myrmekioderma rea (Laubenfels, 1934) (Desmoxyidae) and Topsentia ophiraphidites (Laubenfels, 1934) (Halichondriidae), collected from 30 to 184 m depths, were recorded for the first time from State of Maranhão, north-northeast coast of Brazilian shelf.