Role of model organisms and nanocompounds in human health risk assessment.

Safeguarding the environment is one of the most serious modern challenges, as increasing amounts of chemical compounds are produced and released into the environment, causing a serious threat to the future health of the Earth as well as organisms and humans on a global scale. Ecotoxicology is an integrative science involving different physical, chemical, biological, and social aspects concerned with the study of toxic effects caused by natural or synthetic pollutants on any constituents of ecosystems, including animals (including humans), plants, or microorganisms, in an integral context. In recent decades, this science has undergone considerable development by addressing environmental risk assessments through the biomonitoring of indicator species using biomarkers, model organisms, and nanocompounds in toxicological assays. Since a single taxon cannot be representative of complex ecotoxicological effects and mechanisms of action of a chemical, the use of test batteries is widely accepted in ecotoxicology. Test batteries include properly chosen organisms that are easy to breed, adapt easily to laboratory conditions, and are representative of the environmental compartment under consideration. One of the main issues of toxicological and ecotoxicological research is to gain a deeper understanding of how data should be obtained through laboratory and field approaches using experimental models and how they could be extrapolated to humans. There is a tendency to replace animal tests with in vitro systems and to perform them according to standardized analytical methods and the rules of the so-called good laboratory practice (GLP). This paper aims to review this topic to stimulate both efforts to understand the toxicological and ecotoxicological properties of natural and synthetic chemicals and the possible use of such data for application to humans.

Modulatory Activities of Plant Extracts on Jellyfish Cytotoxicity.

INTRODUCTION: The potential efficacy of selected plant extracts to counteract the dermal toxicity of jellyfish envenomation was investigated using an in vitro cell culture model. METHODS: We studied plant extracts from Carica papaya, Ananas comosus, and Bouvardia ternifolia, known for their antivenom properties, in pairwise combinations with tissue homogenates of the jellyfish Pelagia noctiluca, Phyllorhiza punctata, and Cassiopea andromeda, to evaluate modulations of jellyfish cytotoxic effects. L929 mouse fibroblasts were incubated with pairwise jellyfish/plant extract combinations and examined by MTT assay (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide). RESULTS: C papaya and A comosus significantly lowered the cytotoxicity of P noctiluca and P punctata but induced a slight worsening of C andromeda cytotoxicity. Conversely, B ternifolia was protective against P punctata, ineffective against P noctiluca, and worsened C andromeda cytotoxicity. CONCLUSIONS: Data showed species-specific and contrasting effects of plant extracts, suggesting that those containing protease activities, namely A comosus and C papaya, are more effective in lowering the cytotoxicity of jellyfish venom containing toxic peptidic factors such as phospholipase A. However, all examined plants require further investigation in vivo to evaluate their ability to counteract jellyfish injury to the skin.

Stress-Induced Mucus Secretion and Its Composition by a Combination of Proteomics and Metabolomics of the Jellyfish Aurelia coerulea.

BACKGROUND: Jellyfish respond quickly to external stress that stimulates mucus secretion as a defense. Neither the composition of secreted mucus nor the process of secretion are well understood. METHODS: Aurelia coerulea jellyfish were stimulated by removing them from environmental seawater. Secreted mucus and tissue samples were then collected within 60 min, and analyzed by a combination of proteomics and metabolomics using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS), respectively. RESULTS: Two phases of sample collection displayed a quick decrease in volume, followed by a gradual increase. A total of 2421 and 1208 proteins were identified in tissue homogenate and secreted mucus, respectively. Gene Ontology (GO) analysis showed that the mucus-enriched proteins are mainly located in extracellular or membrane-associated regions, while the tissue-enriched proteins are distributed throughout intracellular compartments. Tryptamine, among 16 different metabolites, increased with the largest-fold change value of 7.8 in mucus, which is consistent with its involvement in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway 'tryptophan metabolism'. We identified 11 metalloproteinases, four serpins, three superoxide dismutases and three complements, and their presence was speculated to be related to self-protective defense. CONCLUSIONS: Our results provide a composition profile of proteins and metabolites in stress-induced mucus and tissue homogenate of A. coerulea. This provides insight for the ongoing endeavors to discover novel bioactive compounds. The large increase of tryptamine in mucus may indicate a strong stress response when jellyfish were taken out of seawater and the active self-protective components such as enzymes, serpins and complements potentially play a key role in innate immunity of jellyfish.

Antimicrobials from Cnidarians. A New Perspective for Anti-Infective Therapy?

The ability of microbes to counter the scientific and therapeutic advancements achieved during the second half of the twentieth century to provide effective disease treatments is currently a significant challenge for researchers in biology and medicine. The discovery of antibiotics, and the subsequent development of synthetic antimicrobial compounds, altered our therapeutic approach towards infectious diseases, and improved the quality and length of life for humans and other organisms. The current alarming rise in cases of antibiotic-resistance has forced biomedical researchers to explore new ways to recognize and/or produce new antimicrobials or to find other approaches for existing therapeutics. Aquatic organisms are known to be a source of compounds having the potential to play a role in fighting the battle against pathogenic microbes. In this connection, cnidarians occupy a pre-eminent role. Over the past few decades several studies have explored the antimicrobial/antibiotic properties of cnidarian extracts with the aim of isolating compounds possessing useful therapeutic features. This paper aims to review the existing data on this subject, taking into account the possible utilization of identified compounds.

Assessment of the Antimicrobial Activity of Algae Extracts on Bacteria Responsible of External Otitis.

External otitis is a diffuse inflammation around the external auditory canal and auricle, which is often occurred by microbial infection. This disease is generally treated using antibiotics, but the frequent occurrence of antibiotic resistance requires the development of new antibiotic agents. In this context, unexplored bioactive natural candidates could be a chance for the production of targeted drugs provided with antimicrobial activity. In this paper, microbial pathogens were isolated from patients with external otitis using ear swabs for over one year, and the antimicrobial activity of the two methanol extracts from selected marine (Dunaliella salina) and freshwater (Pseudokirchneriella subcapitata) microalgae was tested on the isolated pathogens. Totally, 114 bacterial and 11 fungal strains were isolated, of which Staphylococcus spp. (28.8%) and Pseudomonas aeruginosa (P. aeruginosa) (24.8%) were the major pathogens. Only three Staphylococcus aureus (S. aureus) strains and 11 coagulase-negative Staphylococci showed resistance to methicillin. The two algal extracts showed interesting antimicrobial properties, which mostly inhibited the growth of isolated S. aureus, P. aeruginosa, Escherichia coli, and Klebsiella spp. with MICs range of 1.4 × 108 to 2.2 × 10(10) cells/mL. These results suggest that the two algae have potential as resources for the development of antimicrobial agents.

Ecotoxicological assessment of the micelle encapsulator F-500.

Surfactants are synthetic chemicals utilized as detergents and cleaning products or as dispersants and emulsifiers to face water pollution. In spite of this, due to their wide diffusion, surfactants can induce water and soil pollution, notably in developed countries, and can be toxic to organisms. Taking into account that the assessment of new compounds is mandatory in the European Union, in this research the ecotoxicity of fire-fighting micelle encapsulator F-500, newly utilized as dispersant in seawaters polluted with oil dumping, was evaluated. The assessment was carried out on a battery of test-organisms (freshwater algae, crustaceans, and larval fish; seawater algae, crustaceans, and bivalves; soil earthworms, and seeds) as well as on cultured cells (L-929 mouse fibroblasts), which were exposed to F-500 concentrations. According to the toxicity thresholds provided by GESAMP, F-500 resulted to be slightly or moderately toxic to all test-organisms, excluding the freshwater alga Pseudokirchneriella subcapitata that suffered highly toxic effects with IC50 values ranging from 0.21 to 0.49mg/L. The IC50 for mouse fibroblasts was 5.41µg/L after 24h treatment.

Mediterranean jellyfish venoms: a review on scyphomedusae.

The production of natural toxins is an interesting aspect, which characterizes the physiology and the ecology of a number of marine species that use them for defence/offence purposes. Cnidarians are of particular concern from this point of view; their venoms are contained in specialized structures--the nematocysts--which, after mechanical or chemical stimulation, inject the venom in the prey or in the attacker. Cnidarian stinging is a serious health problem for humans in the zones where extremely venomous jellyfish or anemones are common, such as in temperate and tropical oceanic waters and particularly along several Pacific coasts, and severe cases of envenomation, including also lethal cases mainly induced by cubomedusae, were reported. On the contrary, in the Mediterranean region the problem of jellyfish stings is quite modest, even though they can have anyhow an impact on public health and be of importance from the ecological and economic point of view owing to the implications on ecosystems and on some human activities such as tourism, bathing and fishing. This paper reviews the knowledge about the various aspects related to the occurrence and the stinging of the Mediterranean scyphozoan jellyfish as well as the activity of their venoms.

Nematocyst types and venom effects of Aurelia aurita and Velella velella from the Mediterranean Sea.

Natural substances produced by venomous marine organisms are thought to be possible sources of useful compounds and new drugs having the potential to open new ways for pharmacology, nutrition and environmental applications. In this framework, cnidarians are very interesting being widely distributed and all are venomous organisms; so, a deep knowledge of their occurrence, morphology of venomous structures and of effects of venoms at cellular level is fundamental to evaluate the possible utilization of venomous compounds or extracts. In this research, the morphology and occurrence of nematocysts in two cnidarian species (Aurelia aurita, Velella velella), and the preliminary evaluation of the cytotoxicity of V. velella crude extract, of which cytotoxicity on cell cultures at present is unknown, were considered. The specimens were sampled in Güllük Bay, Southwestern coast of Turkey, and in the Gulf of Genova, Northwestern coast of Italy. Six nematocyst types (a-isorhiza, A-isorhiza, O-isorhiza, eurytele, polyspiras, birhopaloid) having different sizes, were observed in A. aurita, and two types (eurytele and stenotele) in V. velella. The crude extract from V. velella showed cytotoxic activity against cultured fibroblasts L929 at high doses, while inducing cell proliferation at low doses. The protein content in the extract increased remarkably after disruption of nematocysts.

Natural Compounds and Drug Discovery: Can Cnidarian Venom Play a Role?

Natural compounds extracted from organisms and microorganisms are an important resource for the development of drugs and bioactive molecules. Many such compounds have made valuable contributions in diverse fields such as human health, pharmaceutics and industrial applications. Presently, however, research on investigating natural compounds from marine organisms is scarce. This is somewhat surprising considering that the marine environment makes a major contribution to Earth's ecosystems and consequently possesses a vast storehouse of diverse marine species. Interestingly, of the marine bioactive natural compounds identified to date, many are venoms, coming from Cnidarians (jellyfish, sea anemones, corals). Cnidarians are therefore particularly interesting marine species, producing important biological compounds that warrant further investigation for their development as possible therapeutic agents. From an experimental aspect, this review aims to emphasize and update the current scientific knowledge reported on selected biological activity (antiinflammatory, antimicrobial, antitumoral, anticoagulant, along with several less studied effects) of Cnidarian venoms/extracts, highlighting potential aspects for ongoing research towards their utilization in human therapeutic approaches.

Comparative proteomic analysis of Aurelia coerulea for its locomotion system molecular structure-function inference.

BACKGROUND: Rhythmic contraction and autonomous movement play a key role in the predation, production and displacement of jellyfish. METHODS: Four independent body parts of the jellyfish Aurelia coerulea, including Bell, Tentacle, Oral arm and Gastric pouch were extracted and have been carried out a compared proteomics by liquid chromatography-mass spectrometry/mass-spectrometry (LC-MS/MS). ResultsA total of 13,429 peptides and 1916 proteins with molecular weights in the range of 10.6-980.9 kDa were identified, where 1916, 1562, 1474 and 1441 proteins were matched in the Gastric pouch, Tentacle, Oral arm and Bell, respectively. Gene Ontology (GO) analysis showed that translation, cytoplasma and ATP binding occupy the top differential terms of the three subdomains Biological process, Cellular Component and Molecular Function. A total of 326 pathways were successfully mapped that are mainly associated with intracellular synthesis, metabolism as well as intracellular functions. Moreover, a total of 27 contractile machinery associated proteins including 22 myosin, 3 actin and 2 tropomyosin were identified. CONCLUSIONS: Our results provide a composition profile in the four independent body parts of the jellyfish A. coerulea, of which the identified muscular proteins will greatly help in the understanding of the structural and functional relationship, as well as their operating mechanisms in the jellyfish locomotion system. SIGNIFICANCE: Omics studies have gained a new overall insight into the function of gene and protein networks during the development of motor systems in both bilateral and radial symmetrical animals. A compared proteomics using the label-free method of nano-LC-MS/MS has been performed through the four independent body parts of the moon jellyfish A. coerulea, including Bell, Tentacle, Oral arm and Gastric pouch. In addition to conventional bioinformatics analyses such as GO and KEGG, we have scanned the locomotion-related components, aligned their sequences, simulated three dimensional structures as well as did the molecular phylogenetic analyses. Our investigation provides a composition profile in the four independent body parts of the jellyfish A. coerulea, of which the identified muscular proteins will greatly help in the understanding of the structural and functional relationship, as well as their operating mechanisms in the jellyfish locomotion system.

The mauve stinger Pelagia noctiluca (Forsskål, 1775). Distribution, ecology, toxicity and epidemiology of stings. A review.

The toxicity of Cnidaria is a subject of concern due to its influence on humans. In particular, jellyfish blooms can highly affect human economical activities, such as bathing, fishery, tourism, etc., as well as the public health. Stinging structures of Cnidaria (nematocysts) produce remarkable effects on human skin, such as erythema, swelling, burning and vesicles, and at times further severe dermonecrotic, cardio- and neurotoxic effects, which are particularly dangerous in sensitive subjects. In several zones the toxicity of jellyfish is a very important health problem, thus it has stimulated the research on these organisms; to date toxicological research on Cnidarian venoms in the Mediterranean region is not well developed due to the weak poisonousness of venoms of jellyfish and anemones living in this area. In spite of this, during last decades several problems were also caused in the Mediterranean by stinging consequent to Cnidarian blooms mainly caused by Pelagia noctiluca (Forsskål, 1775) which is known to be the most venomous Mediterranean jellyfish. This paper reviews the knowledge on this jellyfish species, particularly considering its occurrence and toxicity.

Adverse Health Effects of Betel Quid and the Risk of Oral and Pharyngeal Cancers.

Global reports estimate 600 million betel quid (BQ) chewers. BQ chewing has been demonstrated not only to be a risk factor for cancers of the oral cavity and pharynx and oral potentially malignant disorders (OPMD) but also to cause other cancers and adverse health effects. Herein, we summarized the international comparison data to aid in the understanding of the close relationship between the prevalence of BQ chewing, the occurrence of oral and pharyngeal cancers, and adverse health effects. Potential biomarkers of BQ carcinogens, such as areca nut, alkaloids, and 3-methylnitrosaminopropionitrile (MNPN), are closely associated with human health toxicology. Molecular mechanisms or pathways involving autophagy, hypoxia, COX-2, NF-κB activity, and stemness are known to be induced by BQ ingredients and are very closely related to the carcinogenesis of cancers of oral and pharynx. BQ abuse-related monoamine oxidase (MAO) gene was associated with the occurrence and progress of oral and pharyngeal cancers. In summary, our review article provides important insights into the potential roles of environmental BQ (specific alkaloid biomarkers and nitrosamine products MNPN) and genetic factors (MAO) and offers a basis for studies aiming to reduce or eliminate BQ-related OPMD and oral/pharyngeal cancer incidences in the future.

Indolealkylamines from Toad Vertebrates and Sea Invertebrates - Their Identification and Potential Activities on the Central Nervous System.

Indolealkylamines (IAAs) are biogenic amines and derivatives of 5-hydroxytryptamine, acting primarily on serotonin receptors. IAAs are often considered the most thoroughly investigated group of aromatic amines in the amphibian skin. On the contrary, at present the detailed knowledge of these compounds in lower organisms is still limited and the biogenic amine receptors, mediating hormonal and modulatory functions, are largely unknown in primitive invertebrates. However, some active research is currently underway investigating this class of biogenic amines. Notably, during the last three decades several investigations have demonstrated the biological activity of endogenous biogenic amines in cnidarians, which are known to be the lowest beings equipped with an effective, even though rudimentary, nervous system. Toads, especially those from the Bufonidae family, constitute a significant part of the amphibian family and are an identified source of IAAs. To date fourteen IAAs have been identified in the skins of toad species. All are 5-substituted IAA derivatives acting mainly on the central nervous system (CNS), with most exhibiting some degrees of 5-HT2A receptor selectivity. This selective ability presents potential for their use in the development of treatments for various disorders such as schizophrenia, depression, anxiety, obsessive-compulsive disorders and chronic pain conditions. There are indications that some IAAs may also show subclass selectivity through binding to multiple 5-HT receptor subtypes. Thus, there exists an additional promising platform for the development of therapeutics targeting multiple 5-HT receptors. In this review, IAAs occurring naturally in various species of toad skins, which have been identified and isolated since 1944 are summarized and comparisons are made with similar biogenic amines recognized in cnidarians to date. Such comparisons highlight the potential to utilize existing knowledge gathered from vertebrates, such as toads in order to improve the understanding of the activities of such compounds in lower invertebrates.

Mesozooplankton assemblage and first record of Paracartia grani Sars G.O., 1904 (Copepoda: Calanoida) in the Western harbour of Genova (Ligurian Sea).

Harbours are characterized by high pollutant charge and by the occurrence of well adapted and resistant species. This paper reports the results of an annual plankton survey (May 1997-June 1998) carried out in the western harbour of Genova (Ligurian Sea) and in its mouth. Plankton samples were collected by horizontal trawls using a WP2 net. Copepods were the bulk of plankton in almost all samples. Eight copepod species were recognized: Paracalanus parvus and Acartia clausi were the most abundant. The first record of Paracartia grani in the harbour of Genova is here reported; this species, which is known to occur in polluted harbour waters of the Mediterranean Sea and was found in semi-confined Mediterranean and Atlantic coastal areas, was dominant during October 1997. Also Clausocalanus spp., Centropages typicus, Oithona helgolandica, Oithona nana, Farranula spp., Eurytemora spp., Isias clavipes and Lucicutia spp. were frequently sampled. Among other zooplankters, cladocerans, ostracods and tunicates occurred frequently, while cnidarians, mysids and chaetognaths showed low densities. These results show the occurrence of a well defined harbour plankton and point out the differences between harbour and neritic plankton of the Gulf of Genova.

Ecology of planktonic heterotrophic flagellates. A review.

In aquatic environments heterotrophic flagellates are an important component within the microbial loop and the food web, owing to their involvement in the energy transfer and flux and as an intermediate link between bacteria and primary producers, and greater organisms, such as other protists and metazoan consumers. In the microbial loop heterotrophic flagellates highly contribute to fast biomass and nutrient recycling and to the production in aquatic environments. In fact, these protists consume efficiently viruses, bacteria, cyanobacteria and picophytoplankton, and are grazed mainly by other protists, rotifers and small crustaceans. In this paper the knowledge about these unicellular organisms is reviewed, taking into particular account their ecological relationships and trophic role within the plankton community of marine and freshwater environments.

Animal age-, dose- and cell line-dependent growth of human neuroblastoma in nude mice. A statistical analysis.

Cells lines from human neuroblastoma (NB) and T/lymphoma (T-L) were injected subcutaneously (sc) in female CD1 nu/nu athymic nude mice. Results obtained after the observation of tumour growth were statistically analyzed by SAS. The following four parameters were considered: 1) dose of injected cells, 2) type of injected tumour (NB or T-L), 3) age of mice after individuation of three groups of animals (group A, 4-9 weeks old, group B, 9-20 weeks old, group C, > 20 weeks old), 4) injected cell line within the same tumour type. Latency time (LT), corresponding to the interval between cell inoculum and the appearance of a 5 mm diameter subcutaneous mass, and survival time (ST), corresponding to the interval between cell inoculum and the appearance of a 20 mm diameter subcutaneous tumour mass, were considered to evaluate tumour growth. Results showed that mass progression is affected by the number of injected cells and both LT and ST are age- and dose-dependent; furthermore, significant differences were recorded by using different NB and T-L cell lines. Group C showed longer LT than other groups; group B animals showed a statistically significant longer ST than groups A and C (p < 0.001). Our results indicate that growth of human NB in athymic mice is faster in young animals, which also show a significantly poorer prognosis, while better ST was observed in old and middle-aged animals. Results show statistically significant differences of both LT and ST in animals differing in age and in animals inoculated with different cell amounts. These results seem not to be related with biological properties of NB cells too, since neither the occurrence of MYCN amplification nor chromosome 1p deletion significantly modified such behaviour.

A fast centrifuge method for nematocyst isolation from Pelagia noctiluca Forskal (Cnidaria: Scyphozoa).

Nematocyst isolation from surrounding tissue is an important step to characterize Cnidarian venom. Although several protocols have been used to extract venoms from cnidarian tissues, the complete isolation of nematocysts from tissue is still difficult. The goal of the present work was to evaluate the effectiveness of three different media, Percoll, Ficoll and Methylcellulose in isolating nematocysts from Pelagia noctiluca tentacles by centrifugation. The complete sedimentation of nematocysts and tissue fragments to the bottom of the test tubes was observed in Ficoll and Methylcellulose suspensions. The best result was obtained using a discontinuous density gradient of Percoll: three types of nematocysts were concentrated in three different fractions along the density gradient. Protein assay and preliminary chromatographic analyses confirmed these results.

Hemolytic venoms from marine cnidarian jellyfish - an overview.

Cnidarian jellyfish are viewed as an emergent problem in several coastal zones throughout the world. Recurrent outbreaks pose a serious threat to tourists and bathers, as well as to sea-workers, involving health and economical aspects. As a rule, cnidarian stinging as a consequence of nematocyst firing induces merely local symptoms but cardiovascular or neurological complications can also occur. Hemolysis is a frequent effect of cnidarian stinging; this dangerous condition is known to be caused by several venoms and can sometimes be lethal. At present, the bulk of data concerning hemolytic cnidarian venoms comes from the study of benthic species, such as sea anemones and soft corals, but hemolytic factors were found in venoms of several siphonophore, cubozoan and scyphozoan jellyfish, which are mainly involved in the envenomation of bathers and sea-workers. Therefore, the aim of this paper is to review the scientific literature concerning the hemolytic venoms from cnidarian jellyfish taking into consideration their importance in human pathology as well as health implications and possible therapeutic measures.

The role of Cnidaria in drug discovery. A review on CNS implications and new perspectives.

Many organisms produce bioactive substances used in the production of drugs. In this context, Cnidaria occupy a major position; for this reason, research on new bioactive substances has focused upon them as an interesting target. As a matter of fact, substances and extracts able to fight human diseases have been found in cnidarians, several of which have been studied in laboratories using animal models or cell cultures and, at present, some are in the pre-clinical phase. This review aims to highlight the research on existing drugs or new drug candidates extracted from Cnidaria and the recent patents published in this field; furthermore, as many cnidarian venoms are known to have an impact on the CNS and on neuromuscular transmission, this review particularly considers the research concerning CNS drug discovery and pending patents.

Cytotoxic and cytolytic cnidarian venoms. A review on health implications and possible therapeutic applications.

The toxicity of Cnidaria is a subject of concern for its influence on human activities and public health. During the last decades, the mechanisms of cell injury caused by cnidarian venoms have been studied utilizing extracts from several Cnidaria that have been tested in order to evaluate some fundamental parameters, such as the activity on cell survival, functioning and metabolism, and to improve the knowledge about the mechanisms of action of these compounds. In agreement with the modern tendency aimed to avoid the utilization of living animals in the experiments and to substitute them with in vitro systems, established cell lines or primary cultures have been employed to test cnidarian extracts or derivatives. Several cnidarian venoms have been found to have cytotoxic properties and have been also shown to cause hemolytic effects. Some studied substances have been shown to affect tumour cells and microorganisms, so making cnidarian extracts particularly interesting for their possible therapeutic employment. The review aims to emphasize the up-to-date knowledge about this subject taking in consideration the importance of such venoms in human pathology, the health implications and the possible therapeutic application of these natural compounds.