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Inducible chemical defenses are more common in temperate seaweeds than tropical ones, and are directly detected by increase of chemical contents, or indirectly by differential consumption of live seaweed tissues or artificial food with algal extracts by herbivores. In general, seaweed-induced chemical defense occur between 11 and 20 days after both simulated/artificial or direct herbivory. Here, we used experimental procedures to assess induced chemical defense in the tropical red seaweed Laurencia dendroidea as response to direct grazing, chemical cues from grazed conspecific neighbors and only presence of herbivores. Chemical defenses were analyzed by detecting the palatability of artificial food containing L. dendroidea extracts offered to Aplysia brasiliana and by comparative analyses of extracts from this seaweed by Gas Chromatography/Mass Spectroscopy, as well as metabolomic data analysis by Principal Component Analysis. Our results revealed that direct grazing by A. brasiliana induced a rapid (after 48 h) response among individuals of L. dendroidea, as did waterborne chemical cues from grazed conspecifics, but the presence of sea hare alone did not elicit a response. Increased resistance to grazing was accompanied by significative changes in sesquiterpene metabolomic chemical profile, revealing that induced defense: may be more widespread among seaweeds, independent of latitude; can involve changes in other classes of substances besides phlorotannins or not only the increase in the content of a single compound; and may be a rapid and ecologically coherent response to consumers. In addition, the importance of incorporating the metabolomic approach when examining inducible chemical defense in seaweeds is also emphasized.

The sterol 3β,5α,6β,7α-tetrahydroxyergosta-8(14),22-diene was obtained from bio-guided fractioning of the chloroform extract of 50 L of liquid culture of Acremonium persicinum. This fungal strain was selected because of its anti-proliferative activity against solid human tumour cell lines (GI50 ≤ 50 μg/mL) in a bio-prospective study of fungi isolated from plant material, sediment and water samples obtained from alkaline lakes Alchichica and Atexcac in Puebla, Mexico. This compound showed GI50 (μM) values of: 16, 24, 18, 15 and 12 against tumour cell lines A-549, HBL-100, HeLa, T-47D and WiDr respectively. GI50 effects against tumour lines T-47D and WiDr were found to be greater than the clinically used drugs Etoposide and Cisplatin. Because of this, the results obtained support the pharmacological importance of the microorganisms that develop in these ecosystems and strengthen the non-invasive bio-prospection studies that our work group has developed in recent years.

Background: Fungi from mangrove forests represent a great potential for the bioprospecting of new bioactive metabolites. The rhizosphere of several regions of the world has been widely studied, however, there are no studies on the biodiversity of Mexican mangroves. Objectives: To evaluate the anti-proliferative activity of fungal extracts isolated from accumulated sediments in the rhizosphere of three mangrove species distributed in three zones with different degrees of salinity in Mexico. Methods: The isolated fungi were identified at genus level by taxonomic keys. The anti-proliferative activity was determined by the SRB assay and the strains with values of GI 50

Marine dinoflagellate microalgae belonging to the genus Amphidinium are a key source of an interesting group of polyketide metabolites with potent bioactivities, wide-ranging functional diversity and stereochemical complexity, but low natural availabilities. The feasibility of a microalgae dinoflagellate-based sustainable bioprocess for producing amphidinols (APDNs) by photoautotrophic culture of Amphidinium carterae in a pilot-scale LED-illuminated bubble column photobioreactor (PBR) was therefore investigated. A fed-batch culture mode with pulse feeding strategy provided a growth pattern strongly limited by the availability of phosphate content in the culture medium that stimulated the production of cellular APDNs. Since A. carterae was found to be much more shear-sensitive than other shear-tolerant non-dinoflagellate microalgae, the culture height and air flow rate were established to ensure the absence of damaging levels of hydrodynamic stress. The biomass capacity yielded by the PBR at the end of the culture (0.540 g d.w. L−1 equivalent to 1.70 × 106 cell mL−1) corresponded to that estimated stoichiometrically from the experimentally determined biomass P-molar formula (C329 O126 H732 N69 S3 P1) and the total phosphorus and nitrogen balances. The downstream processing section was initially conceived to recover APDNs excreted by cells into the supernatant. A dry APDN-enriched extract concentration of 49 mg per liter of supernatant was obtained. This purification process led to partitioning of the extract into several fractions and sub-fractions thereof. Only two sub-fractions were studied, yielding thereof highly pure (>95% pure) luteophanol D and lingshuiol A, and a new, roughly purified (>80% pure) APDN, namely amphidinol 20. The percentages of luteophanol D, lingshuiol A and amphidinol 20 by dry weight of the APDN-enriched extract obtained were 1%, 0.39% and 0.31%, respectively, thus representing a concentration in the culture supernatant of 490, 191 and 152 μg L−1, respectively.

Consumption of Brassica (Cruciferae) vegetables is associated with a reduced risk of cancer, but identification of the active components and insights into the underlying molecular events are scarce. Here we found that an extract of Lepidium latifolium, a cruciferous plant native to southern Europe, Mediterranean countries and Asia, showed in vitro cytotoxic activity, inducing caspase-dependent apoptosis, in a variety of human tumor cells, and the plant juice showed in vivo antitumor activity in a HT-29 human colon cancer xenograft mouse model. The epithionitrile 1-cyano-2,3-epithiopropane (CETP) was identified as the major active cancer cell-killing principle of L. latifolium. Synthetic and plant-derived CETP displayed similar proapoptotic activities as assessed by biochemical and morphological analyses. Analysis of the antiproliferative capacity of CETP on a wide number of cancer cell lines from the NCI-60 cell line panel followed by COMPARE analysis, showed an activity profile different from known anticancer agents. Flow cytometry and biochemical analyses revealed that CETP-induced apoptosis involved mitochondria, as assessed by loss of mitochondrial transmembrane potential and generation of reactive oxygen species, while overexpression of Bcl-X and Bcl-2 prevented CETP-induced apoptosis. Inhibition of reactive oxygen species by glutathione and N-acetyl cysteine reduced the apoptotic response induced by CETP. FADD dominant negative form, blocking Fas/CD95 signaling, and a specific caspase-8 inhibitor also inhibited CETP-induced killing. Taken together, our data suggest that the cancer cell-killing action of CETP, involving both intrinsic and extrinsic apoptotic signaling pathways, underlies the antitumor activity of L. latifolium plant, which could be of potential interest in cancer treatment., This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2011-30518; SAF2014-59716-R; CTQ2014-55888-C03-01-R), Instituto de Salud Carlos III (RD12/0036/0065 from Red Temática de Investigación Cooperativa en Cáncer, cofunded by the EU's European Regional Development Fund—FEDER), and the European Community's Seventh Framework Programme FP7-2007-2013 (HEALTH-F2-2011-256986, PANACREAS).

It is well known that marine fungi are an excellent source of biologically active secondary metabolites, and by 2011, it was reported that over 400 bioactive metabolites were derived from marine fungi.This study establishes the basis for future research on antiproliferative compounds of marine endophytes inhabited in the Veracruz Reef System.Isolation of the 34 fungal strains was carried out by microbiological method from samples of sponges, corals, and other biological material from the Veracruz Reef System. The fungal biomass and broth were separated and extracted with a mixture of solvents MeOH:CHCl3. Characterization and molecular identification of the fungal strains were performed through microbiological methods and the analysis of the ITS-rDNA regions. Antiproliferative activity was tested at a dose of 250 μg/mL on human solid tumor cell lines HBL-100, HeLa, SW1573, T-47D, and WiDr by the SRB assay after 48 h-exposure to the fungal extracts.The extracts from five isolates showed an antiproliferative effect against one or more of the tested cell lines (percentage growth50%). The mycelial extract from the isolate LAEE 03 manifested the highest activity against the five cell lines (% PG of 17 HBL-100, 19 HeLa, 23 SW1573, -6 T-47D, and 10 WiDr) and the strain was identified as Curvularia trifolii (Kauffman) Boedijn (Pleosporaceae).The results obtained indicate that the extract from a marine derived C. trifolii has the antiproliferative effect, thus suggesting that this organism is a good candidate for further analysis of its metabolites.

Various species of the genus Ganoderma have been used for centuries according to oriental tradition as a source of medicines and nutrients. A chemical study of the fruiting bodies and mycelial culture of G. oerstedii was carried out with the idea of isolating and characterizing active natural components present to make use of their potential pharmaceutical application in Mexico. The fruiting bodies and mycelial culture of G. oesrtedii were lyophylized and extracted one after the other with hexane, chloroform, and methanol. Following this process, each substance was extracted separately by using column chromatography. From fruiting bodies eight metabolites, five sterols (ergosta-7,22-dien-3β-ol, ergosterol peroxide, ergosterol, cerevisterol, and ergosta-7,22-dien-3-one) as well as three terpene compounds (ganodermanondiol, ganoderic acid Sz, and ganoderitriol M) were obtained from fruiting bodies. From the mycelial culture three metabolites, two sterols (ergosterol and cerevisterol), and a new terpene compound (ganoderic acetate from the acid) were obtained. These structures were established based on a spectroscopic analysis mainly using nuclear magnetic resonance and a comparison with data already established.

Red algae of Laurencia continue to provide wide structural diversity and complexity of halogenated C acetogenin medium-ring ethers. Here, we described the isolation of three new C acetogenins (3–5), and one truncated derivative (6) from Laurencia viridis collected on the Canary Islands. These compounds are interesting variations on the pinnatifidenyne structure that included the first examples of ethynyl oxirane derivatives (3–4). The structures were elucidated by extensive study of NMR (Nuclear Magnetic Resonance) data, J-based configuration analysis and DFT (Density Functional Theory) calculations. Their antiproliferative activity against six human solid tumor cell lines was evaluated., This work was supported by grant CTQ2014-55888-C03-01-R (Ministry of Economy and Competitiveness of Spain, MINECO). A.M.-A. and C.R.d.V. acknowledge MINECO for a Ph.D. scholarship and O.M.-F. thanks ULL for a grant.

8 pages, 1 table, 3 figures, Three strains of the toxic benthic dinoflagellate Prorocentrum hoffmannianum were isolated in the Canary Islands (north-east Atlantic Ocean, Spain). The identity of the strains was determined by phylogenetic analyses of partial LSU rDNA (D1-D2 regions) but their morphology based on SEM images corresponded to P. maculosum. Their toxin profiles were analyzed by liquid chromatography and high resolution mass spectrometry analysis (LC–HRMS) on cell extracts and culture media. Okadaic acid and three analogs were detected in all strains. Rather, in culture media the detected compounds were variable among strains, two of them being okadaic acid analogs not found on cell extracts. As a result, the taxonomy of the species was revised and P. maculosum is proposed as a junior synonym of P. hoffmannianum whose description is emended., This work was funded by projects CICAN, CCVIEO and CTQ2014-55888-C3-01-R

Five-membered rings are of particular interest, due to their presence in some of the most common molecules in chemistry and biology. Despite their apparent simplicity, the structural resolution of these rings is complex, due to their inherent conformational flexibility. Here, we describe an application of a recently reported simple and efficient NMR protocol based on the measurement of spin-spin coupling constants to achieve the challenging relative configurations of five new halogenated C15 tetrahydrofuranyl-acetogenins, marilzafurollenes A–D (1–4) and 12-acetoxy-marilzafurenyne (5), isolated from the red alga, Laurencia marilzae. Although DFT chemical shift calculations were used to connect remote stereocenters, the NMR-based approach seems advantageous over computational techniques in this context, as the presence of halogens may interfere with reliable calculations.

The structural determination of small organic molecules is mainly undertaken by using NMR techniques, although it is increasingly supplemented by using computational methods. NMR parameters, such as chemical shifts and coupling constants, are extremely sensitive indicators of local molecular conformation and are a source of structural evidence. However, their interpretation is fairly challenging in many circumstances, such as the case of the new polyether squalene derivative nivariol, the structure of which was elucidated by means of NMR spectroscopy and DFT calculations. The potential flexibility of this molecule and the high number of quaternary carbon atoms that it contains make its configurational assignment very difficult. Moreover, the relative configuration of four separated stereoclusters was established and subsequently connected by using NOE and J ‐based analysis, as well as by a comparison of its experimental 13C NMR chemical shifts with the corresponding population‐weighted values, as calculated by using DFT methods. Limitations of these used approaches became apparent but were overcome by combining the two methods.

A set of crown ethyl acyl derivatives based on 18-crown-6 moiety was synthesized and evaluated for biological activity. In vitro antiproliferative profiling demonstrated significant activities against HBL-100, HeLa, SW1573 and WiDr human cell lines. The most active compound exhibited GI50 values in the range of 3.7–5.6 μM. Antimicrobial evaluation showed that three polyaromatic compounds were active against Staphylococcus aureus (MIC90 values from 8.3 μM to 50 μM), whereas a (decyloxy)benzene substitution exhibited moderate activity against Candida albicans (MIC90 values 36 μM). According to SAR evaluation, the size of the crown ether and the acyl side chain had a significant effect on the bioactivity. Aromatic moieties close to the acyl group led to improved bioactivity as exemplified by some of the tested compounds. These results provide further evidence on the potential of crown ethyl structure as a scaffold for developing new biological probes and lead candidates for drug development.

Macrosporin, a bioactive metabolite produced by a plant-pathogenic fungus Stemphylium lycopersici , was found to be a non-host selective toxin which plays a part in leaf necrosis, a disease caused by this fungus. Here, we discuss the role of this metabolite in necrotic spots, through the direct inoculation of the midribs of the leaves of a natural host, as well as its ability to generate active oxygen species, through the process of photosensitizing oxygen.

Five‐membered rings are clearly among the most common structural motifs found in chemistry and biology. Nevertheless, the configuration of conformationally mobile five‐membered rings is often difficult to assign from nuclear magnetic resonance (NMR) data. A simple, reliable, and efficient approach for the stereochemical analysis of five‐membered rings based on the measurement of NMR coupling constants is presented. Density functional theory calculations using representative conformations of the full conformational space available to rings with different substitution patterns were used to identify differences between the accessible coupling constant values for cis and trans relative orientations of the substituents. The calculations were assessed experimentally using NMR data obtained from a number of models. This approach can be easily used to analyze different five‐membered rings, such as oxolanes, cyclopentanes, furanosides and pyrrolidines, and their relative configuration can be determined without the need for making further conformational considerations.

Eight new halogenated C(15) acetogenins, 1-8, were isolated from the organic extract of the red alga Laurencia marilzae. The structure elucidation and the assignments of the relative configurations were established by extensive use of spectroscopic studies, particularly 1D and 2D NMR data, while the absolute configurations of compounds 1 and 5 were determined by single-crystal X-ray diffraction analysis. Compounds 1, 2, 4, 5, and 7, along with the previously reported related cyclic ether obtusallene IV (9), were evaluated against six human solid tumor cell lines. All compounds were found to be essentially inactive (GI(50)10 μg/mL).

27 páginas, 4 tablas, 4 figuras, Diarrhetic Shellfish Poisoning (DSP) is a specific type of food poisoning, characterized by severe gastrointestinal illness due to the ingestion of filter feeding bivalves contaminated with a specific suite of toxins. It is known that the problem is worldwide and three chemically different groups of toxins have been historically associated with DSP syndrome: okadaic acid (OA) and dinophysistoxins (DTXs), pectenotoxins (PTXs) and yessotoxins (YTXs). PTXs and YTXs have been considered as DSP toxins because they can be detected with the bioassays used for the toxins of the okadaic acid group, but diarrhegenic effects have only been proven for OA and DTXs. Whereas, some PTXs causes liver necrosis and YTXs damages cardiac muscle after intraperitoneal injection into mice. On the other hand, azaspiracids (AZAs) have never been included in the DSP group, but they cause diarrhoea in humans. This review summarizes the origin, characterization, structure, activity, mechanism of action, clinical symptoms, method for analysis, potential risk, regulation and perspectives of DSP and associated toxins produced by marine dinoflagellates., This work was funded b yGrants AGL2005-07924-C04- 01/02 and CTQ2008-0754-C04-01/04 from the MEC of Spain.

Adriatoxin-B (ATX-B) is the first C13 terminal truncated YTX analogous metabolite that was isolated from large-scale cultures of the dinoflagellate Protoceratium reticulatum. The structure and conformational behavior of ATX-B was elucidated by a combination of NMR spectroscopy experiments and a conformational analysis. The isolation of this metabolite, which possesses a clear structural relationship with adriatoxin, supports the idea that the real source of these cut down substances is in the microalgae and not in the mollusks, via metabolic degradation like it has been assumed until now.

An exclusive club : The first member of an unprecedented class of polyunsaturated and polyhydroxylated macrocycle, belizeanolide (see structure), has been isolated from a marine dinoflagellate. The corresponding open form, belizeanolic acid, was also found. Their structures were determined primarily by analysis of NMR spectroscopic data. Both compounds showed significant antiproliferative activities.

Okadaic acid (1) (OA) and its congeners are mainly responsible for diarrhetic shellfish poisoning (DSP) syndrome. The presence of several OA derivatives have already been confirmed in Prorocentrum and Dinophysis spp. In this paper, we report on the detection and identification of a new DSP toxin, the OA isomer 19-epi-okadaic acid (2) (19-epi-OA), isolated from cultures of Prorocentrum belizeanum, by determining its retention time (RT) and fragmentation pattern using liquid chromatography coupled with mass spectrometry (LC–MS/MS).

In this paper we report on the isolation and structural elucidation of two new noncontiguous polypropionates, micromelones A ( 10) and B ( 11), that have been isolated from the marine gastropod Micromelo undata. Their structures were determined through the interpretation of their spectroscopic data, and a biosynthetic pathway from a common linear precursor with other polypropionate groups has been proposed.

The obtusallenes are a significant subset of C15-halogenated acetogenins that incorporate 12-membered cyclic ethers. We have recently reported the isolation from Laurencia marilzae of 12-epoxyobtusallene IV (1) and its related α,β-unsaturated carboxylate ester (2), both of special biogenetic relevance. Here we describe the final step of our study, the isolation of three new analogues (3-5), among these, the first bromopropargylic derivative (3) of this class of macrocyclic C15-acetogenins. The structures were elucidated by analysis of NMR and X-ray data. 12-Epoxyobtusallene IV (1), its new isomer 4, and known obtusallene IV (6) were evaluated for their apoptosis-inducing activities in a human hepatocarcinoma cell line.

Seven new nonadecaketides (1-7), lobophorols A-C, lobophopyranones A and B, and lobophorones A and B, along with the first naturally occurring related metabolites (8-10), were isolated from specimens of Lobophora variegata collected from the Canary Islands. Their structures were determined by extensive spectroscopic methods. In addition, an insight into the biosynthesis of these compounds on the basis of the involvement of type III polyketide synthases is proposed. Lobophorol A (1) showed significant antibacterial activity against Staphylococcus aureus.

The chemical study of the red alga Laurencia viridis has led to the isolation of four new polyether triterpenoids: 28-hydroxysaiyacenol B (2), saiyacenol C (3), 15,16-epoxythyrsiferol A (4), and 15,16-epoxythyrsiferol B (5). The structures of 2 and 3 were established mainly by NMR data analysis and comparison with the well-known metabolite dehydrothyrsiferol (1). However, due to the existence of a nonprotonated carbon within the epoxide functionality, stereochemical assignments in 4 and 5 required an in-depth structural study that included NOESY data, J-based configuration analysis, comparison with synthetic models, and DFT calculations. The biological activities of the new metabolites and other related oxasqualenoids were evaluated for the first time against a panel of relevant biofouling marine organisms, and structure–activity conclusions were obtained.

Dactylomelol 1 , initially isolated from molluscs of the genus Aplysia , was the first example of a monocarbocyclic diterpene skeleton with a C-6–C-11 cyclisation. This paper reports on the identification and structural elucidation of six new diterpenes, compounds 4 – 9 , isolated from specimens of Laurencia sp., together with dactylomelol, all with this particular carbon backbone that we named dactylomelane. This result is the first to confirm a red alga as the biogenetic origin of this class of compounds.

Marine toxins have drawn wide interest because their economical impact and disastrous effect upon the shellfish industry and public health in many parts of the world. One of the most interesting group of substances of marine toxins, from structural and pharmacological points of view are polyether compounds, which generally present a great diversity in size and potent biological activities. The subject of this work was about to biosynthesis of okadaic acid skeleton as leader as DSP toxins. Its biosynthesis attracts considerable attention since the carbon skeleton has been shown to be synthesised via an unusual route. In this paper we report on stable isotope incorporation experiments on DSP toxin in artificial cultures of dinoflagellate. The comparison of the degrees of incorporation in these samples measured by different methods led to contradictory results. This implies that further experimental data is needed in order to propose a logical biogenetic scheme. © 2004 The Japan Chemical Journal Forum and Wiley Periodicals, Inc. Chem Rec 4: 1–9; 2004: Published online in Wiley InterScience (www.interscience.wiley.com ) DOI 10.1002/tcr.10072

The red seaweed Laurencia viridis is a rich source of secondary metabolites derived from squalene. Novel polyethers, clavidol 4 , 3-epi-1111-dehydrothyrsiferol 5 , and lactodehydrothyrsiferol 6 , have been isolated and their structures established by spectroscopical methods. The relative stereochemistry proposed for these compounds is based on ROESY and NOEDIFF data. Possible biogenetic pathways are also discussed.

Chemical cues from sessile hosts can attract mobile and associated organisms and they are also impotant to maintain associations and overall biodiversity, but the identity and molecular structures of these chemicals have been little explored in the marine environment. Secondary metabolites are recognized as possible chemical mediators in the association between species of Laurencia and Aplysia, but the identity of the compounds has not been established. Here, for the first time, we experimentally verify that the sesquiterpene (+)-elatol, a compound produced by the red seaweed Laurencia dendroidea, is a chemical cue attracting the associated sea hare Aplysia brasiliana. In addition to revealing the nature of the chemical mediation between these two species, we provide evidence of a chemical cue that allows young individuals of A. brasiliana to live in association with L. dendroidea. This study highlights the importance of chemical cues in Laurencia-Aplysia association.

Two new zoantharia-derived metabolites, zoaramine (I) and zoarenone (II), are characterized.

In our search for quorum-sensing (QS) disrupting molecules, 75 fungal isolates were recovered from reef organisms (endophytes), saline lakes and mangrove rhizosphere. Their QS inhibitory activity was evaluated in Chromobacterium violaceum CVO26. Four strains of endophytic fungi stood out for their potent activity at concentrations from 500 to 50 μg mL−1. The molecular characterization, based on the internal transcribed spacer (ITS) region sequences (ITS1, 5.8S and ITS2) between the rRNA of 18S and 28S, identified these strains as belonging to four genera: Sarocladium (LAEE06), Fusarium (LAEE13), Epicoccum (LAEE14), and Khuskia (LAEE21). Interestingly, three came from coral species and two of them came from the same organism, the coral Diploria strigosa. Metabolic profiles obtained by Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) suggest that a combination of fungal secondary metabolites and fatty acids could be the responsible for the observed activities. The LC-HRMS analysis also revealed the presence of potentially new secondary metabolites. This is, to the best of our knowledge, the first report of QS inhibition by marine endophytic fungi.

Belizentrin (1), a novel 25-membered polyketide-derived macrocycle, was isolated from cultures of the marine dinoflagellate Prorocentrum belizeanum. This metabolite is the first member of an unprecedented class of polyunsaturated and polyhydroxylated macrolactams. The structure of 1 was primarily determined by NMR and computational methods. Pharmacological assays with cerebellar cells showed that 1 produces important changes in neuronal network integrity at nanomolar concentrations.

Three new norzoanthamine-type alkaloids, named 2-hydroxy-11-ketonorzoan thamide B (1), norzoanthamide B (2) and 15-hydroxynorzoanthamine (3), were isolated from Zoanthus sp. specimens collected at the Canary Islands. Their structures were determined by interpretation of NMR and HR-ESIMS data. Relative configurations of their chiral centers were proposed on the basis of ROESY spectra and by comparison of their spectroscopic data with those of the well-known compound, norzoanthamine.

Four new triterpenoids with a squalene carbon skeleton martiriol 7, pseudodehydrothyrsiferol 8, dioxepandehydrothyrsiferol 9 and 16-epihydroxydehydrothyrsiferol 10 have been isolated from the red alga Laurencia viridis. The structures were determined through the interpretation of their spectral data and the relative stereochemistries were proposed on the basis of ROESY and NOEDIFF data. Their cytotoxic activities were evaluated.

A new monogalactosyl triacylglycerol has been isolated from the artificial cultures of the dinoflagellate Amphidinium sp. Its structure was determined through the interpretation of 2D-NMR spectral data and chemical degradation. The relative steieochemistry is proposed on the basis of ROESY data.

Three new metabolites have been isolated from the brown alga Cystoseira abies marina. the structures were established by using spectroscopic methods.