RISPOSTA IMMUNITARIA DEL PUNTERUOLO ROSSO NEI CONFRONTI DEL ENTOMOPATOGENO BACILLUS THURINGIENSIS

Introdotto accidentalmente in Sicilia nel 2005 Rhynchophorous ferrugineus Olivier (Coleoptera Curculionidae), noto come punteruolo rosso, ha infestato velocemente e gravemente molte palme. La sua espansione è veloce e i potenziali stress abiotici e biotici presenti nel nuovo areale non sembrano costituire dei possibili agenti di contenimento. Si è cercato di comprendere il perché di tale resistenza trattando larve di punteruolo con uno dei maggiori batteri entomopageni attualmente conosciuti il Bacillus thuringiensis Berliner (Bt). Gli studi eff ettuati hanno evidenziato una certa mortalità ma soprattutto un’interazione con l’attività trofi ca. Per comprendere meglio questi eff etti si è st…

FACIT collagen (1alpha-chain) is expressed by hemocytes and epidermis during the inflammatory response of the ascidian Ciona intestinalis

Based on previous cloning and sequencing study, real-time PCR and in situ hybridization assays of the inflamed body wall of LPS-injected Ciona intestinalis showed the enhanced gene expression of a collagen with FACIT structural features (Ci-type IX-Col 1a-chain). By using specific antibodies raised against an opportunely chosen Ci-type IX-Col synthetic peptide, the fibroblast property of hemocytes challenged in vitro with LPS (at 4 h) was displayed by flow cytometry, while immunocytochemistry identified hemocytes with large granules (morula cells) as collagen-producing cells. Hemocyte lysate supernatant analyzed in immunoblotting contained a 60 kDa band identifiable as 1a-chain-Ci-type IX-C…

Physiological advantages of dwarfing in surviving extinctions in high-CO2 oceans

Excessive CO 2 in the present-day ocean-atmosphere system is causing ocean acidification, and is likely to cause a severe biodiversity decline in the future, mirroring effects in many past mass extinctions. Fossil records demonstrate that organisms surviving such events were often smaller than those before, a phenomenon called the Lilliput effect. Here, we show that two gastropod species adapted to acidified seawater at shallow-water CO 2 seeps were smaller than those found in normal pH conditions and had higher mass-specific energy consumption but significantly lower whole-animal metabolic energy demand. These physiological changes allowed the animals to maintain calcification and to parti…

Preliminary evidence for the cytotoxic molecule in hemocyte supernatant lysate from Ciona intestinalis unilocular hemocytes

METHYLMERCURY EFFECTS ON Mytilus galloprovincialis HAEMOCYTES ACTIVITY

Bivalves, filter-feeding organisms, due to their wide distri-bution, are used by many countries in biomonitoring, particu-larly in the assessment of xenobiotics in the marine ecosystem.These sentinel organisms are able to accumulate trace metalsand other substances in their tissues. Haemocytes are effectorsof cellular immunity in bivalves and are capable of respond-ing to stressors through activities such as phagocytosis or cyto-toxicity. In order to identify cellular markers to investigate pol-lution, the effect of different concentrations of organic mercury(CH3HgCl) on the morphology and responses of Mytilus gal-loprovincialishaemocytes was studied in this work. Sublethalconcentrations of…

Galectine nella risposta infiammatoria di Ciona intestinalis cDNA ed espressione genica

Inflammatory-like reaction following bacterial injection and antimicrobial peptide isolation from Anemonia sulcata (Cnidaria)

Antimicrobial response in Anemonia sulcata (Cnidaria)

Putative rhamnose-binding lectin in the solitary ascidian Ciona intestinalis.

INFLAMMATION AND TENTACLE REGENERATION RESPONSES IN ANEMONIA VIRIDIS (ANTHOZOA, CNIDARIA)

Isolation, characterization and expression analysis of a collectin in Tunicate Ciona intestinalis.

Isolation of a novel LPS-induced component of the ML superfamily in Ciona intestinalis

ML superfamily represents a group of proteins playing important roles in lipid metabolism and innate immune response. In this study, we report the identification of the first component of the ML superfamily in the invertebrate Ciona intestinalis by means of a subtractive hybridization strategy. Sequence homology and phylogenetic analysis showed that this protein forms a specific clade with vertebrate components of the Niemann-Pick type C2 protein and, for this reason, it has been named Ci-NPC2. The putative Ci-NPC2 is a 150 amino acids long protein with a short signal peptide, seven cysteine residues, three putative lipid binding site and a three-dimensional model showing a characteristic b…

Attività antibatterica di peptidi estratti da celomociti di echinodermi

Negli echinodermi la difesa dell’organismo dalle invasioni microbiche avviene tramite meccanismi cellulari e umorali dell’immunità innata. Una componente fondamentale dei fattori umorali è costituita dai peptidi antimicrobici (AMPs) molto conservati con un ampio spettro di attività antimicrobica contro batteri, virus e funghi. Solitamente gli AMPs sono costituiti da meno di 100 amminoacidi, sono cationici e formano strutture anfipatiche. In questo lavoro dimostriamo che i celomociti dell’Echinoideo Paracentrotus lividus e dell’Oloturoideo Holothuria tubulosa contengono sostanze con attività antimicrobica verso diversi ceppi batterici patogeni umani. In entrambe le specie, nel precipitato ac…

LPS injection reprograms the expression and the 3′ UTR of a CAP gene by alternative polyadenylation and the formation of a GAIT element in Ciona intestinalis

The diversification of cellular functions is one of the major characteristics of multicellular organisms which allow cells to modulate their gene expression, leading to the formation of transcripts and proteins with different functions and concentrations in response to different stimuli. CAP genes represent a widespread family of proteins belonging to the cysteine-rich secretory protein, antigen 5 and pathogenesis-related 1 superfamily which, it has been proposed, play key roles in the infection process and the modulation of immune responses in host animals. The ascidian Ciona intestinalis represents a group of proto-chordates with an exclusively innate immune system that has been widely st…

Components of hemocyte extracts from marine invertebrates exert antimicrobial activity

Ciona intestinalis galectin (CiLgals-a and CiLgals-b) genes are differentially expressed in endostyle zones and challenged by LPS

Abstract Immunohistochemical and in situ hybridization assays were performed to answer the question whether the endostyle, that is the initial gastro-intestinal trait of Ciona intestinalis pharynx, is involved in galectin (CiLgals-a and CiLgals-b) production during the pharynx inflammatory response to LPS inoculation. Specific anti-CiLgal-a and anti-CiLgals-b antibodies, and oligonucleotide probes, that mark inflammatory hemocytes inside the pharynx vessels and vessel epithelium as shown by a previous paper, were assayed on endostyle histological sections. For the first time, we show that galectins are produced by endostyle zones, and both CiLgals-a and –b genes are upregulated by LPS. CiLg…

Cellule che producono ciTNFalfa sono attive nell'infiammazione e nello sviluppo larvale di Ciona intestinalis

A D-galactose specific lectin is an inducible inflammatory IL-1-like opsonin in the hemolymph of the ascidian Ciona intestinalis challegend with LPS.

Attivazione del sistema della profenolossidasi nella risposta infiammatoria della tunica di Ciona intestinalis (L.)

The Expression of an Immune-Related Phenoloxidase Gene is Modulated in Ciona Intestinalis Ovary, Test Cells, Embryos and Larva

Two distinct Ciona intestinalis phenoloxidases (CinPO1, 2) had previously been cloned and sequenced. The CinPO2 is involved in innate immunity and is expressed by inflammatory hemocytes that populate the tunic and pharynx vessels as a response to LPS inoculation. In situ hybridization and immunohistochemistry assays on histological section, showed that the expression of this gene and the produced protein are shared with oogenesis, embryogenesis and larval morphogenesis. Intriguingly, upregulation of gene transcription was found in the test cell layer that envelopes the ovary follicle, ovulated egg, and gastrula, as well as it was modulated in the zygotic nucleus of outer balstomers of 32-ce…

Role of mucosal immune response and histopathological study in European eel (Anguilla anguilla L.) intraperitoneal challenged by Vibrio anguillarum or Tenacibaculum soleae.

Abstract The external mucus layer that covers fish skin contains numerous immune substances scarcely studied that act as the first line of defence against a broad spectrum of pathogens. This study aimed to characterize and describe for the first time several humoral immune defence parameters in the skin mucus of the European eel (Anguilla anguilla) after intraperitoneal injection with Vibrio anguillarum or Tenacibaculum soleae. This study evaluated several immune-related enzymes and bactericidal activity against fish pathogenic bacteria in the skin mucus of European eels at 24, 48, and 72 h post-challenge. The results demonstrated that European eel skin mucus showed significant increments i…

RIFLESSIONI SULL'EDUCAZIONE SCIENTIFICA MEDIANTE L'INSEGNAMENTO DELLE SCIENZE DELLA VITA

Cnidarian Interaction with Microbial Communities: From Aid to Animal’s Health to Rejection Responses

The phylum Cnidaria is an ancient branch in the tree of metazoans. Several species exert a remarkable longevity, suggesting the existence of a developed and consistent defense mechanism of the innate immunity capable to overcome the potential repeated exposure to microbial pathogenic agents. Increasing evidence indicates that the innate immune system in Cnidarians is not only involved in the disruption of harmful microorganisms, but also is crucial in structuring tissue-associated microbial communities that are essential components of the Cnidarian holobiont and useful to the animal’s health for several functions, including metabolism, immune defense, development, and behavior. Someti…

Mesoglea Extracellular Matrix Reorganization during Regenerative Process in Anemonia viridis (Forskål, 1775)

Given the anatomical simplicity and the extraordinary ability to regenerate missing parts of the body, Cnidaria represent an excellent model for the study of the mechanisms regulating regenerative processes. They possess the mesoglea, an amorphous and practically acellular extracellular matrix (ECM) located between the epidermis and the gastrodermis of the body and tentacles and consists of the same molecules present in the ECM of vertebrates, such as collagen, laminin, fibronectin and proteoglycans. This feature makes cnidarians anthozoans valid models for understanding the ECM role during regenerative processes. Indeed, it is now clear that its role in animal tissues is not just tissue su…

Study of immunotoxicity responses of Sabella spallanzanii exposed to copper sulphate

The Inflammatory Response of Urochordata: The Basic Process of the Ascidians’ Innate Immunity

Ascidians form a widespread marine invertebrate group and are heterogeneous in terms of the taxonomic groups’ evolutionary lineages. The ascidian genomes lack significant homologies for rearranging genes of the vertebrate adoptive immunity. Genome analysis, gene sequencing, and transcriptional profiling have allowed us to disclose upregulation of innate immunity genes and cell labeling with riboprobes and antibodies has identified hemocyte types in tunic and pharynx inflammatory responses. Lymphocyte-like cells are stem cells and their immunocompetence has been proposed. Granulocyte types (compartment/morula cells) and hemocytes with large granules/vacuoles (compartment/morula cells) are ma…

Lectine sieriche di "tipo F" nell'immunità innata dei pesci: aspetti molecolari e funzionali.

Hemocytes of Rhynchophorus ferrugineus (Olivier)(Coleoptera: Curculionidae) and their response to Saccharomyces cerevisiae and Bacillus thuringiensis

Originally from tropical Asia, the Red Palm Weevil (RPW Rhynchophorus ferrugineus (Olivier) is the most dangerous and deadly pest of many palm trees, and there have been reports of its recent detection in France, Greece and Italy. At present, emphasis is on the development of integrated pest management based on biological control rather than on chemical insecticides, however the success of both systems is often insufficient. In this regard, RPW appears to be one pest that is very difficult to control. Thus inves- tigations into the natural defences of this curculionid are advisable. RPW hemocytes, the main immuno- competent cells in the insect, are described for the first time. We identifie…

Purification and characterization of D-galactose binding lectin involved in the inflammatory response in Ciona intestinalis

Ciona intestinalis interleukin 17-like genes expression is upregulated by LPS challenge

In humans, IL-17 is a proinflammatory cytokine that plays a key role in the clearance of extracellular bacteria promoting cell infiltration and production of several cytokines and chemokines. Here, we report on three Ciona intestinalis IL-17 homologues (CiIL17-1, CiIL17-2, CiIL17-3). The gene organisation, phylogenetic tree and modelling supported the close relationship with the mammalian IL-17A and IL-17F suggesting that the C. intestinalis IL-17 genes share a common ancestor in the chordate lineages. Real time PCR analysis showed a prompt expression induced by LPS inoculation suggesting that they are involved in the first phase of inflammatory response. In situ hybridization assays disclo…

Upregulated transcription of phenoloxidase genes in the pharynx and endostyle of Ciona intestinalis in response to LPS

We investigated the role of phenoloxidases (POs) in ascidians inflammatory reaction, a components of a copper-containing protein family involved in invertebrate immune system. In Ciona intestinalis two phenoloxidases (CinPO-1, CinPO-2) have been sequenced. In the present study, real time PCR analysis showed that both CinPO-1 and CinPO-2 genes were modulated by LPS inoculation suggesting that they are inducible and highly expressed in the inflamed pharynx. In situ hybridization disclosed CinPO-1 and CinPO-2 transcripts in pharynx hemocytes (granulocytes) and, mainly, in unilocular refractile granulocytes (URG) which mainly populated the inflamed tunic matrix. Interestingly, the genes are als…

Further insight on Ciona intestinalis prophenoloxidase system activated during the LPS induced inflammatory response

A lytic mechanism based on soluble phospholypases A2 (sPLA2) and b-galactoside specific lectins is exerted by Ciona intestinalis (ascidian) unilocular refractile hemocytes against K562 cell line and mammalian erythrocytes

Abstract Hemocytes from the ascidian Ciona intestinalis exert in vitro Ca 2+ -dependent cytotoxic activity toward mammalian erythrocytes and K562 cells. To examine the lytic mechanism, hemocyte populations were separated (B1–B6 bands) through a Percoll discontinuous density gradient, the hemocyte cytotoxic activity (HCA) and the lytic activity of the hemocyte lysate supernatant (HLS) were assayed. In addition the separated hemocytes were cultured and the cell-free culture medium (CFM) assayed after 3 h culture. Results support that unilocular refractile hemocytes (URGs), enriched in B5, are cytotoxic. The B5-HLS contains lysins and the activity of B5-CFM shows that lysins can be released in…

Distinct protoconchs recognised in three of the larger Mediterranean Cerithium species (Caenogastropoda: Cerithiidae)

The gastropod genus Cerithium includes several polymorphic species which are hardly distinguishable using a morphological approach based on teleoconch characters. Here we show that protoconch characters can be reliably used to identify the larger Mediterranean species (Cerithium alucastrum, C. repandum and C. vulgatum), and to assess their intraspecific variability. Based on a large amount of morphological data, we show that a multispiral, strongly sculptured protoconch (traditionally associated with C. vulgatum) is found in C. alucastrum. This species originated in the Pliocene. A multispiral, weakly sculptured protoconch, not observed previously, is reported for C. vulgatum. A paucispiral…

Serum lectins in fish innate immunity: molecular and functional aspects

Fucose-binding lectins (FBL) are present in tissues and fluids from invertebrates and vertebrates. The lectin repertoires in teleost fish are highly diversified and recently has been described the structure of the fucose-binding agglutinin that revealed a novel lectin fold (the “F-type” eel (Anguilla anguilla) fold), which shared a unique fucose-binding sequence motif contained both in carbohydrate-binding proteins and unrelated proteins. In this report, we describe serum FBL from sea bass Dicentrarchus labrax and sea bream Sparus aurata. These lectins were purified, characterized, cloned and sequenced. Studies on structural aspects, biological activity, tissue distribution as well as ontog…

Effetti dell’esposizione a cadmio e rame sull’attività di cellule del sangue di spigola (Dicentrarchus labrax L. Osteichthyes, Moronidae).

From Cnidarian immunobiology to cultural heritage applications

The study of cnidarians immunity, as model systems of metazoans, lead additional informations on the first steps of the immunity evolution. The functions of the genes and cellular pathways in higher vertebrates are conserved during the evolution of metazoans, as shown by the discovery of homologues in cnidarians. These basal metazoans in fact, are far from "simples" in the range of methods at their disposal to deal with potential prey but also invading microbes and pathogens. They can give informations about the invertebrates innate immune repertoire. We investigated the immunobiology starting from the inflammatory response in Anemonia sulcata (Cnidaria: Anthozoa) following injection of sub…

Two similar new species of Alvania Risso,1826(Caenogastropoda: Rissoidae)from the late Cenozoic of Italy

The prophenoloxidase system is activated during the tunic inflammatory reaction of Ciona intestinalis

Phenoloxidase (PO) activity was examined in the tunic tissue of Ciona intestinalis following lipopolysaccharide (LPS) intratunic injection. Tunic homogenate supernatant (THS), assayed with the Dopa-MBTH reaction, displayed Ca(2+)-independent PO activity that was raised by LPS and further enhanced by proteases. Specific inhibitors (tropolone, phenylthiourea, diethylthiocarbamate) supported the specificity of the reaction. Assay with soybean trypsin inhibitor showed that, in the tunic, PO activation with trypsin was not significantly inhibited suggesting that proteases diverse from serine proteases were involved. In vivo experiments were carried out by injecting isosmotic medium or LPS, and T…

Phenoloxidases in ascidian hemocytes: characterization of the pro-phenoloxidase activating system.

The phenoloxidase (PO) activity of the hemocytes lysate supernatant from three ascidians species, assayed by means of 3-methyl-2-benzothiazolinone hydrazone hydrochloride, have been compared. PO-containing hemocytes were identified by a cytochemical reaction and the enzymatic activity measured by a spectrophotometric assay of lysate supernatant from hemocyte populations separated on a discontinuous Percoll density gradient. In Styela plicata, the enzyme appeared to be contained in morula cells only. In Ciona intestinalis, PO activity was shown in univacuolar refractile granulocyte and granular hemocyte. In Phallusia mammillata both compartment cell and granular hemocytes were positive. Enzy…

Invertebrate lectins present cytokine properties.

Inflamed adult pharynx tissues and swimming larva of Ciona intestinalis share CiTNFalfa-producing cells

Invertebrate lectins present cytockine properties.

Biological activity, tissue distribution and preliminary moleular characterization of a serum fucolectin from the sea bass (Dicentrarchus labrax)

Attività citotossica dei celomociti di Holothuria tubulosa (Echinodermata)

Numerosi studi hanno accertato che nel fluido celomatico degli echinodermi sono presenti cellule con attività diretta verso agenti estranei. In particolare i celomociti svolgono un repertorio di funzioni immunitarie tra cui la fagocitosi, la citotossicità, l’attività antibatterica, la formazione di capsule e il rigetto dei trapianti. Nel presente lavoro abbiamo studiato l’attività citotossica dei celomociti di Holothuria tubulosa (Oloturoideo) non separati ed abbiamo utilizzato cellule di mammifero come target. I celomociti prelevati dal fluido celomatico lisano gli eritrociti di coniglio o di montone e le cellule tumorali della linea eritromieloide umana K562. Inoltre anche il supernatante…

Enhanced expression of a cloned and sequenced Ciona intestinalis TNFa-like (CiTNFa) gene during the LPS-induced inflammatory response.

A tumor necrosis factor-alpha (TNFalpha)-like gene from Ciona intestinalis (CiTNF alpha-like) body wall challenged with bacterial lipopolysaccharide (LPS) was cloned and sequenced 4 h after LPS inoculation. An open reading frame of 936 bp encoding a propeptide of 312 amino acids (35.4 kDa) displaying a transmembrane domain from positions 7 to 29, a TACE cleavage site, and a mature peptide domain of 185 amino acids (20.9 kDa), was determined with a predicted isoelectric point of 9.4. The phylogenetic tree based on deduced amino acid sequences of invertebrate TNF-like protein and vertebrate TNFs supported the divergence between the ascidian and vertebrate TNF families, whereas D. melanogaster…

Nuovi dati sulla risposta infiammatoria dell’ascidia Ciona intestinalis

A response of Rhynchophorous ferrugineus (Coleoptera: Curculionidae) larval hemocytes to Bacillus thuringiensis

In the ovary of Ciona intestinalis (Type A), immune-related galectin and phenoloxidase genes are differentially expressed by the follicle accessory cells

Abstract Riboprobes (in situ hybridization) and antibodies (immunohistochemistry), previously used to show the upregulation of Ciona intestinalis (Type A) galectins (CiLgals-a, CiLgals-b) and phenoloxidase (CinPO2) immune-related genes, were tested on histological sections of the ovary. The ovarian follicles are composed of oocytes encased by follicular cells (FCs) and test cells (TCs). Results show the transcription upregulation of both CiLgals and CinPO2 genes in the vitellogenic FCs, conversely distinct cytolocalization of the proteins are shown. At vitellogenic stage, the CiLgals are localized in the FCs, in the oocyte cytoplasm, and close to the germinal vesicle (GV), whereas the CinPO…

Tunicate immunocytes can be cytotoxic toward foreign cells

Sphingomyelin as well as carbohydrates are involved in the mechanism of cytotoxic molecules contained and released in vitro by Ciona intestinalis granulocytes.

Il “problem posing” come metodologia innovativa per lo studio delle Scienze della Vita

Cytotoxic activity of Ciona intestinalis (Tunicata) hemocytes: Properties of the in vitro reaction against erythrocyte targets

Hemocytes (effectors) of Ciona intestinalis showed a natural cytotoxic capacity (HCA) when assayed in vitro against erythrocytes (targets). Cytotoxic cells lysed, to a variable extent, rabbit (RE), human (A, B, O), guinea pig, and sheep (SE) erythrocytes. Hemocyte cytotoxic activity (HCA) assayed against SE is a calcium-dependent reaction, occurs rapidly (15-30 min), at 25-37 degrees C over a wide range of pH (5.4-8.0). Assays were carried out using: 1) the medium in which hemocytes were maintained, 2) the soluble portion of hemocyte lysates, and 3) debris prepared from hemocyte lysates. Results suggest that HCA is a cell-mediated process that requires effector-target cell contacts. Anti-SE…

Il sistema della profenolossidasi in Ciona intestinalis. Caratterizzazione e interazione tra fenolossidasi, perossinectina e superossido dismutasi Cu-Zn dipendente nella risposta infiammatoria indotta da LPS

The expression of an immune-related phenoloxidase gene is modulated inCiona intestinalisovary, test cells, embryos and larva

Two distinct Ciona intestinalis phenoloxidases (CinPO1, 2) had previously been cloned and sequenced. The CinPO2 is involved in innate immunity and is expressed by inflammatory hemocytes that populate the tunic and pharynx vessels as a response to LPS inoculation. In situ hybridization and immunohistochemistry assays on histological section, showed that the expression of this gene and the produced protein are shared with oogenesis, embryogenesis and larval morphogenesis. Intriguingly, upregulation of gene transcription was found in the test cell layer that envelopes the ovary follicle, ovulated egg, and gastrula, as well as it was modulated in the zygotic nucleus of outer balstomers of 32-ce…

Effect of exposure cadmium on the echinoderm Paracentrotus lividus (Echinoidea)

EVOLUTION, ADAPTATION AND IMMUNE FUNCTIONS OF FISH LECTINS

Fish are equipped with a complex lectin repertoire that, like mammals, are involved almost all the immune reactions. Carbohydrate recognition and interactions mediated by lectins have been recognized involved in vertebrate innate immunity, not only for recognition of potential pathogens, but also acting in the agglutination, immobilization and other functional steps. In fish, C, F types galectins, Rhamnose-bind- ing lectin (RBL) and pentraxin have been identified in both car- tilaginous and bony fish. In addition, selectins and other genes have been found in the currently available fish genomes. On the basis of our results about F-type and RBL lectins we showed that: lectin repertoires in f…

Parametri sierologici e cellulari della specie alloctona Procambarus clarckii

Procambarus clarkii è un animale d’acqua dolce originario del Centro-Sud degli Stati Uniti e del Nord-Est del Messico. Questa specie rientra nella lista delle 100 specie invasive più pericolose al mondo (Delivering Alien Invasive Species Inventories for Europe) in quanto presenta una serie di caratteristiche biologiche, che la rendono una minaccia per la biodiversità. In Sicilia la sua presenza è stata registrata per la prima volta nel 2003 nella riserva naturale “Lago Preola e Gorghi Tondi”, Mazara del Vallo (TP). Allo scopo di conoscere meglio la biologia di questa specie, su esemplari di P. clarkii provenienti dalla riserva di Trapani, abbiamo esaminato i tipi emocitari ed alcune attivit…

Modulation of prophenoloxidase activity in Ciona intestinalis inflammatory response

Effetti di Bacillus thuringiensis nella sua forma vegetativa sulle larve di Rhynchophorous ferrugineus (Coleoptera Curculionidae)

Il batterio Bacillus thuringiensis (Bt) è un patogeno di molti insetti ed è ampiamente impiegato, sottoforma di spore, nella lotta biologica. La forma vegetative (Btv) è considerata quella responsabile della setticemia dell’ospite dopo che le tossine (Cry) rilasciate dal corpo parasporale del Bt in forma sporulata (Bts) hanno perforato il mesenteron del ospite. Tuttavia poco è conosciuto relativamente ai meccanismi di difesa dell’insetto nelle fasi successive alla azione delle tossine Cry. Lo scopo del presente lavoro è quello di studiare gli effetti del Btv sulle larve del fitofago Rhynchophorous ferrugineus Olivier (Coleoptera: Curculionidae) a seguito di iniezioni sub letali di Btv. Ques…

A new Miocene deep-sea chiton and early evidence for Teredinidae-sustained wood-fall communities

Deep-sea wood-falls are important biodiversity hot spots for insights on chemosynthesis-based communities. The study of deep-sea wood-fall-related palaeocommunities from the Neogene of north Italy shed light on interesting associations from the Miocene of Torrente Cinghio (Tortonian) and of Moncasale di Casina (Langhian). The most common components of this association are typical chemosynthetic/wood-fall molluscs, such as the gastropods Homalopoma sp. and Pseudonina bellardii, the bivalves Idas sp. and shipworms, and the chiton Leptochiton lignatilis n. sp., which belongs to a genus typical of recent sunken woods in tropical waters. The new species described is compared with other fossil an…

Ciona robusta hemocyte populational dynamics and PO-dependent cytotoxic activity

Hemocyte populations from the ascidian Ciona robusta, separated through a Percoll discontinuous density gradient, are further characterized by May-Grünwald-Giemsa staining and a cytochemical reaction for phenoloxidase. Variability in cell density, acidophilic property and phenoloxidase activity suggest multiple hemocyte type populations, cell lineages and morphotypes that may be involved in distinct cellular responses. Therefore, unilocular refractile granulocytes, typical of this ascidian species, enriched in a fraction separated from the hemolymph show in vitro phenoloxidase-dependent cytotoxic activity against mammalian erythrocytes and a tumor cell lineage, in addition the properties li…

Effects of organic mercury on Mytilus galloprovincialis hemocyte function and morphology

Abstract Filter-feeding organisms accumulate xenobiotics and other substances in their tissues. They can be useful as sentinel organisms in biomonitoring of the marine compartment. Bivalve cellular immunity is ensured by phagocytosis and cytotoxic reactions carried out by hemocytes in a network with humoral responses. These can be affected by chemical contaminants in water that can be immunosuppressors also at a low concentration increasing the sensibility to pathogens. This work is an attempt to individuate cellular markers for pollution detection, investigating the effect of methylmercury (CH3HgCl) at different concentrations on the activity and hemocyte morphology of the Mediterranean m…

Paracentrin 1, a synthetic antimicrobial peptide from the sea-urchin Paracentrotus lividus, interferes with staphylococcal and Pseudomonas aeruginosa biofilm formation

The rise of antibiotic-resistance as well as the reduction of investments by pharmaceutical companies in the development of new antibiotics have stimulated the investigation for alternative strategies to conventional antibiotics. Many antimicrobial peptides show a high specificity for prokaryotes and a low toxicity for eukaryotic cells and, due to their mode of action the development of resistance is considered unlikely. We recently characterized an antimicrobial peptide that was called Paracentrin 1 from the 5-kDa peptide fraction from the coelomocyte cytosol of the Paracentrotus lividus. In this study, the chemically synthesized Paracentrin 1, was tested for its antimicrobial and antibiof…

Inducible galectins are expressed in the inflamed pharynx of the ascidian Ciona intestinalis

Although ascidians belong to a key group in chordate phylogenesis, amino acid sequences of Ciona intestinalis galectin-CRDs (CiLgals-a and -b) have been retained too divergent from vertebrate galectins. In the present paper, to contribute in disclosing Bi-CRD galectin evolution a novel attempt was carried out on CiLgals-a and -b CRDs phylogenetic analysis, and their involvement in ascidian inflammatory responses was shown. CiLgals resulted aligned with Bi-CRD galectins from vertebrates (Xenopus tropicalis, Gallus gallus, Mus musculus, Homo sapiens), cephalochordates (Branchiostoma floridae), echinoderms (Strongylocentrotus purpuratus) and a mono-CRD galectin from the ascidian Clavelina pict…

The conservation and diversity of ascidian cells and molecules involved in the inflammatory reaction: The Ciona robusta model

Ascidians are marine invertebrate chordates belonging to the earliest branch (Tunicata) in the chordate phylum, therefore, they are of interest for studying the evolution of immune systems. Due to the known genome, the non-colonial Ciona robusta, previously considered to be C. intestinalis type A, is a model species for the study of inflammatory response. The internal defense of ascidians mainly relies on hemocytes circulating in the hemolymph and pharynx. Hemocytes can be in vivo challenged by LPS injection and various granulocyte and vacuolated cell populations differentiated to produce and release inflammatory factors. Molecular biology and gene expression studies revealed complex defens…

The cytotoxic activity of Ciona intestinalis (ascidian) unilocular refractile hemocytes versus K562 tumor cells and mammalian erythrocytes involves phospholypase A2 and lectins

F-type lectin from the sea bass (Dicentrarchus labrax): purification, cDNA cloning, tissue expression and localization, and opsonic activity.

Recently described biochemical and structural aspects of fucose-binding lectins from the European eel (Anguilla anguilla) and striped bass (Morone saxatilis) led to the identification of a novel lectin family ("F-type" lectins) characterized by a unique sequence motif and a characteristic structural fold. The F-type fold is shared not only with other members of this lectin family, but also with apparently unrelated proteins ranging from prokaryotes to vertebrates. Here we describe the purification, biochemical and molecular properties, and the opsonic activity of an F-type lectin (DlFBL) isolated from sea bass (Dicentrarchus labrax) serum. DlFBL exhibits two tandemly arranged carbohydrate-r…

Effect of in vitro exposure to cadmiumand copper on sea bass blood cells

Blood cells freshly collected from sea bass (Dicentrarchus labrax) were exposed in vitro to different concentrations of cadmium (Cd) and copper (Cu) at 10-7 M, 10-5 M, 10-3 M, and exam- ined for neutral red retention capacity and for cell vitality with MTT assay. A relationship between heavy metal exposure and alteration in responses of blood cells in a dose-time-dependent was found. Our results showed that fish blood cells may constitute an interesting biological model for experimen- tal and applied toxicology, especially in the case of environmental pollution.

Inflamed adult pharynx tissues and swimming larva of Ciona intestinalis share CiTNFalpha-producing cells.

In situ hybridisation and immunohistochemistry analyses have shown that the Ciona intestinalis tumour necrosis factor alpha gene (CiTNFalpha), which has been previously cloned and sequenced, is expressed either during the inflammatory pharynx response to lipopolysaccharide (LPS) or during the swimming larval phase of development. Granulocytes with large granules and compartment/morula cells are CiTNFalpha-producing cells in both inflamed pharynx and larvae. Pharynx vessel endothelium also takes part in the inflammatory response. Haemocyte nodules in the vessel lumen or associated with the endothelium suggest the involvement of CiTNFalpha in recruiting lymphocyte-like cells and promoting the…

Localizzazione di una lectina di “tipo F” nell’adulto e nell’ontogenesi di Dicentrarchus labrax

The purification, cloning, sequencing, molecular properties and expression of a fucose-binding lectin from the serum of Dicentrarchus labrax (DlFBL) have been previously reported. We now describe the distribution and expression of DlFBL during fish ontogeny. Immunohistochemistry and in situ hybridization assays were carried out at various developmental stages (from 10 days posthatching larvae to juveniles). Another fucose-binding lectin, similar to DlFBL in biochemical, immunochemical and agglutinating properties, was extracted and purified from eggs and appeared to be localized in the embryo yolk sack residual. DlFBL was found in columnar and goblet cells of the intestinal epithelium of la…

Molecular cloning, characterization and expression analysis of peroxinectin from Ciona intestinalis

LPS challenge regulates gene expression and tissue localization of a Ciona intestinalis gene through an alternative polyadenylation mechanism

Isolation and characterization of a LPS induced MD2-like protein in Ciona intestinalis

The MD2 (Myeloid Differentiation factor-2) protein belongs to the ML superfamily. This group of proteins contain a specific lipid binding domain (ML domain) that plays an important role in lipid recognition and metabolism. In vertebrates, MD-2 is involved in innate immune response as co-receptor in the LPS/TLR4 signaling pathway; MD2 recognizes and binds the bacterial lipid A and drives the TLR4 activation. Two TLR isoforms, CiTLR-1 and CiTLR-2, were identified in Ciona intestinalis with a TIR domain most similar to human TLR4 and TLR 6 respectively. Using a PCR-based subtractive hybridization strategy for isolation of differentially expressed genes between LPS-challenged and naïve C. intes…

Ciona intestinalis peroxinectin is a novel component of the peroxidase–cyclooxygenase gene superfamily upregulated by LPS

Peroxinectins function as hemoperoxidase and cell adhesion factor involved in invertebrate immune reaction. In this study, the ascidian (Ciona intestinalis) peroxinectin gene (CiPxt) and its expression during the inflammatory response have been examined. CiPxt is a new member of the peroxidase-cyclooxygenase gene superfamily that contains both the peroxidase domain and the integrin KGD (Lys-Gly-Asp) binding motif. A phylogenetic tree showed that CiPxt is very close to the chordate group and appears to be the outgroup of mammalian MPO, EPO and TPO clades. The CiPxt molecular structure model resulted superimposable to the human myeloperoxidase. The CiPxt mRNA expression is upregulated by LPS …

AMPs and biotechnology application for new generation of medical devices

Chemiluminescence response of ß-glucan stimulated leukocytes isolated from different tissues and peritoneal cavity of Dicentrarchus labrax

Abstract The respiratory burst of leukocytes isolated from sea bass ( Dicentrarchus labrax ) pronephros, peritoneal cavity (P.C.), spleen and blood, was measured by a chemiluminescence (CL) assay after stimulation with β-glucan. The CL response by P.C. and pronephros leukocytes was significantly higher than that expressed by a similar number of cells separated from spleen and blood. This probably reflects the observation that the proportion of macrophages and neutrophils was highest in the populations of leukocytes from peritoneal cavity and pronephros. Comparative observations showed a higher degree of yeast phagocytosis by leukocytes taken from peritoneal cavity than the pronephros. Moreo…

Transforming growth factor β (CiTGF-β) gene expression is induced in the inflammatory reaction of Ciona intestinalis.

Transforming growth factor (TGF-β) is a well-known component of a regulatory cytokines superfamily that has pleiotropic functions in a broad range of cell types and is involved, in vertebrates, in numerous physiological and pathological processes. In the current study, we report on Ciona intestinalis molecular characterisation and expression of a transforming growth factor β homologue (CiTGF-β). The gene organisation, phylogenetic tree and modelling supported the close relationship with the mammalian TGF suggesting that the C. intestinalis TGF-β gene shares a common ancestor in the chordate lineages. Functionally, real-time PCR analysis showed that CiTGF-β was transcriptionally upregulated …

Encapsulation Response ofCiona intestinalis(Ascidiacea) to Intratunical Erythrocyte Injection

Previous studies on the ascidian Ciona intestinalis have shown that an encapsulation response is experimentally induced by inserting vertebrate erythrocytes into the tunic, which initiates a massive inflammatory cell infiltration to isolate the injured area. Several hemocytes contribute to capsule formation, destruction of the foreign cells, tunic regeneration, and wound healing. The fine features of some inflammatory cell types are described although the complete capsular structure is not yet reported. Accordingly, the present investigation further examines various aspects of this cellular reaction against erythrocytes and, for the first time, presents the involvement of extratunical circu…

F-type lectin from serum of the Antarctic teleost fish Trematomus bernacchii (Boulenger, 1902): Purification, structural characterization, and bacterial agglutinating activity.

Abstract The increasing availability of sequenced genomes has enabled a deeper understanding of the complexity of fish lectin repertoires involved in early development and immune recognition. The teleost fucose-type lectin (FTL) family includes proteins that preferentially bind fucose and display tandemly arrayed carbohydrate-recognition domains (CRDs) or are found in mosaic combinations with other domains. They function as opsonins, promoting phagocytosis and the clearance of microbial pathogens. The Antarctic fish Trematomus bernacchii is a Perciforme living at extremely low temperatures (−1.68 °C) which is considered a model for studying adaptability to the variability of environmental w…

Expression of a type IX-like collagen in tissue injury of the ascidian Ciona intestinalis durino inflammatory process

Cnidarian Immunity and the Repertoire of Defense Mechanisms in Anthozoans

Anthozoa is the most specious class of the phylum Cnidaria that is phylogenetically basal within the Metazoa. It is an interesting group for studying the evolution of mutualisms and immunity, for despite their morphological simplicity, Anthozoans are unexpectedly immunologically complex, with large genomes and gene families similar to those of the Bilateria. Evidence indicates that the Anthozoan innate immune system is not only involved in the disruption of harmful microorganisms, but is also crucial in structuring tissue-associated microbial communities that are essential components of the cnidarian holobiont and useful to the animal’s health for several functions including metabolism, imm…

Studio preliminare sulla caratterizzazione degli emociti di Cancer pagurus

specific inflammatory response of Anemonia sulcata (cnidaria) after bacterial injection cause tissue rejection and enzymatic activity alteration

The evolution of multicellular organisms was marked by adaptation to protect against pathogens. The mechanisms for discriminating the ''self'' from ''non-self” have evolved into a long history of cellular and molecular strategies from damage repair to the co-evolution of host-pathogen interaction. The phylum of Cnidaria is one of the first branches in the tree of animal life to provide crucial insights on the evolution of immunity. Sea anemones (Anthozoa, Cnidaria) are benthic sessile species able to maintain the integrity of the tissues and allorecognition in colonial forms and to differentiate between symbionts and pathogenic intruders. We investigated the inflammatory response in sea ane…

Il sistema della profenolossidasi nella risposta infiammatoria della tunica di Ciona intestinalis (L.)

Effetti del cadmio sulle attività cellulari dell’echinoderma Paracentrotus lividus (Echinoidea)

Attività biologica, distribuzione tissutale e caratterizzazione molecolare della fucolectina sierica di spigola (Dicentrarchus labrax).

Encapsulation Response ofCiona intestinalis(Ascidiacea) to Intratunical Erythrocyte Injection

Abstract Electron microscopic studies on the encapsulation induced by erythrocyte injection into the tunic of the ascidian Ciona intestinalis were carried out. The observations reported in the present paper complete the description previously given of capsule architecture and contribute to the characterization of the cells involved in the inflammatory reaction. The inflamed area is surrounded by an ample and peculiar “three-layered coat” respectively composed of flattened and packed extratunical hemocytes, the monolayered epithelium, and a layer of intratunical electron-dense particles. The latter are also clustered, variously arranged, and distributed in the tunic ground substance. The epi…

An Attempt to re-examine the immune role of Ciona intestinalis hemocytes

Evoluzione dell’immunità innata. Componenti della reazione infiammatoria indotta da LPS nella parete corporea dell’ascidia Ciona intestinalis.

Effect of Bacillus thuringiensis as vegetative form on hemocytes of Rhynchophorous ferrugineus (Coleoptera Curculionidae) larvae

The bacterium Bacillus thuringiensis (Bt) is a pathogen of many insect species and is actively used in biocontrol. The vegetative form as been reported to be involved in insect septicemia process. Bt during the vegetative stage of growth, is known to secern a new family of insecticidal proteins. Moreover recently evidence has been provided, that B. thuringiensis can establish itself in replicative and vegetative form on the leaf surface. Little is known on the interaction of pathogens with the defense responses of phytophagous insects. Insect circulating hemocytes are primarily responsible for the immune defense against parasites and pathogens. We use as model Rhynchophorous ferrugineus Oli…

The Ciona intestinalis immune-related galectin genes (CiLgals-a and CiLgals-b) are expressed by the gastric epithelium.

The transcription of two Ciona intestinalis galectin genes (CiLgals-a and CiLgalseb) is uparegulated by LPS in the pharynxis (hemocytes, vessel epithelium, endostilar zones) which is retained the main organ of the immunity. In this ascidian, for the first time we show, by immunohistochemistry and in situ hybridization methods, that these two immune-related genes are expressed in the gastric epithelium of naïve ascidians, whereas the galectins appear to be only contained in the intestine columnar epithelium. In addition, according to previous results on the pharynx, the genes are also expressed and galectins produced by hemocytes scattered in the connective tissue surrounding the gut. The ge…

Reazione infiammatoria ed isolamento di un peptide antimicrobico da Anemonia sulcata (Cnidaria)

Espressione di galactine nell'infiammazione e nella larva natante di Ciona intestinalis

Enhanced CiTNFα expression is an inflammatory response of the ascidian Ciona intestinalis

Il sistema della profenolossidasi in Ciona intestinalis durante il processo infiammatorio indotto da LPS

The Ciona intestinalis prophenoloxidase activating system during LPS inflammatory reaction

Cloning and expression of a novel component of the CAP superfamily enhanced in the inflammatory response to LPS of the ascidian Ciona intestinalis.

The CAP superfamily is a group of proteins that have been linked to several biological functions such as reproduction, cancer, and immune defense. A differential screening between lipopolysaccharide (LPS)-challenged and naive Ciona intestinalis has been performed to identify LPS-induced genes. This strategy has allowed the isolation of a full-length 1471-bp cDNA encoding for a 413-amino-acid protein (CiCAP). In silico analysis has shown that this polypeptide displays a modular structure with similarities to vertebrate CAP-superfamily proteins and to a collagen-binding adhesin of Streptococcus mutans. Domain organization analysis and alignment of CiCAP to other vertebrate CAP proteins have r…

Physiological and agonistic behavioural response of Procambarus clarkii to an acoustic stimulus

Summary This study examined the effects of an acoustic stimulus on the haemolymph and agonistic behaviour of the red swamp crayfish Procambarus clarkii. The experiment was conducted in a tank equipped with a video recording system using 6 groups (3 control and 3 test groups) of five adult crayfish (30 specimens in total). After one hour of habituation, the behaviour of the crayfish was monitored for two hours. During the second hour, the animals in the test groups were exposed to a linear sweep (frequency range 0.1-25 kHz; peak amplitude 148 dBrms re 1 µPa at 12 kHz) acoustic stimulus for 30 minutes. Exposure to the noise produced significant variations in haemato-immunological parameters a…

FACIT collagen (1α-chain) is expressed by hemocytes and epidermis during the inflammatory response of the ascidian Ciona intestinalis

Based on previous cloning and sequencing study, real-time PCR and in situ hybridization assays of the inflamed body wall of LPS-injected Ciona intestinalis showed the enhanced gene expression of a collagen with FACIT structural features (Ci-type IX-Col 1alpha-chain). By using specific antibodies raised against an opportunely chosen Ci-type IX-Col synthetic peptide, the fibroblast property of hemocytes challenged in vitro with LPS (at 4h) was displayed by flow cytometry, while immunocytochemistry identified hemocytes with large granules (morula cells) as collagen-producing cells. Hemocyte lysate supernatant analyzed in immunoblotting contained a 60 kDa band identifiable as 1alpha-chain-Ci-ty…

cytokines in the inflammatory response of the ascidian ciona intestinalis

The Ciona intestinalis inflammatory response to several irritants have been demonstrated to be composed of a complex set reaction. The cellular reactions involve hemocyte infiltration, hemocyte and epidermis activities, vacuolization, cell disruption, while cell products can contribute to form capsule components and/or cause a wound. In this response the involvement of the pharynx, as the main immune-competent organ, has been disclosed by lipopolysaccharide (LPS) challenge that upregulates innate immunity genes. In vertebrate cytokines modulate the balance between humoral and cell-based immune responses, and they regulate the maturation, growth, and responsiveness of cell populations that h…

Modificazioni indotte dal metilmercurio sugli emociti dell’ascidia Styela plicata

Nuovi dati sull'attivazione della profenolossidasi di Ciona intestinalis nel processo infiammatorio

Prophenoloxidase system is activated in the tunic inflammatory response of ciona intestinalis

Cell cooperation in coelomocyte cytotoxic activity of Paracentrotus lividus coelomocytes

The coelomic fluid from the sea urchin Paracentrotus lividus contains several coelomocyte types including amoebocytes and uncoloured spherulocytes involved in immune defences. In the present paper, we show a Ca(2+)-dependent cytotoxic activity for the unfractionated coelomocytes assayed in vitro, with rabbit erythrocytes and the K562 tumour cell line. In a plaque-forming assay, whole coelomocyte preparations as well as density gradient separated coelomocyte populations revealed that cell populations enriched in uncoloured spherulocytes, exerted high cytotoxic activity by releasing lysins in the presence of amoebocytes. This cooperative effect could be dependent on soluble factors released b…

Molecules and cells in inflammatory responses of the ascidian Ciona intestinalis

Taxonomy and palaeobiogeography of the Cenozoic Euro-Mediterranean rissoid gastropod Galeodinopsis and its relationship with close genera

The investigation of the Late Paleogene to Late Neogene species of rissoid gastropod Galeodinopsis in the Euro-Mediterranean area has supported the hypothesis that this genus is an intermediate form between two well-known rissoids, Alvinia and Manzonia. We recognized four species of Galeodinopsis: G. biangulata, G. germanica sp. nov., G. semperi (new name for Rissoa duboisii), and G. tiberiana. The oldest (very Late Eocene/Oligocene) representatives of Galeodinopsis, G. biangulata, and G. semperi, share similar shell shape and microsculpture with Alvinia. This suggests that Galeodinopsis originated from some Eocene species related to Alvinia. The new species represents the first occurrence …

Interleukin 17 genes as mediators of inflammatory responses in Ciona intestinalis.

Inflammation is a complex reaction of host defence mechanisms aiming at neutralization of an insult and restoring normal tissue structure and function. In human IL-17 is T-cell derived cytokine plays a key role in the clearance of extracellular bacteria promoting cell infiltration and production of several cytokines and chemokines. Here, we report on three Ciona intestinalis IL-17 homologues (CiIL17-1, CiIL17-2, CiIL17-3). The gene organization, phylogenetic tree and modeling supported the close relationship with the mammalian IL-17A and IL-17F suggesting that the C. intestinalis IL-17 genes share a common ancestor in the chordate lineages. Real time PCR analysis showed a prompt expression …

La risposta infiammatoria in Ciona intestinalis è caratterizzata dall’aumento di lectine specifiche per D-galattosidi e dall’espressione di collagene FACIT di tipo IX

Comparative analysis of fucose binding lectins isolated and characterized from different teleost species, and distribution of a F-Lectin during Dicentrarchus labrax ontogenesis

EFFETTI DELLE VARIAZIONI STAGIONALI IN DUE POPOLAZIONI DI ANEMONIA SULCATA (PENNANT, 1777) E PRIME OSSERVAZIONI SULLA RIGENERAZIONE DEI TENTACOLI

ALESSANDRO LENTINI, MARIA GIOVANNA PARISI, DANIELA PARRINELLO, MATTEO CAMMARATA Dipartimento di Scienze della Terra e del Mare, Viale delle Scienze ed. 16 Università di Palermo EFFETTI DELLE VARIAZIONI STAGIONALI IN DUE POPOLAZIONI DI ANEMONIA SULCATA (PENNANT, 1777) E PRIME OSSERVAZIONI SULLA RIGENERAZIONE DEI TENTACOLI Gli organismi bentonici sono utilizzati sia come indicatori delle condizioni oceanografiche che come strumenti per il monitoraggio delle condizioni della fascia marino costiera su cui insistono fattori antropici. La raccolta di informazioni di base sui meccanismi di acclimatazione stagionale è un passo fondamentale per comprendere le risposte fisiologiche degli indicatori b…

Specific inflammatory response of Anemonia sulcata (Cnidaria) after bacterial injection causes tissue reaction and enzymatic activity alteration

The evolution of multicellular organisms was marked by adaptations to protect against pathogens. The mechanisms for discriminating the ’’self’’ from ’’non-self” have evolved into a long history of cellular and molecular strategies, from damage repair to the co-evolution of host-pathogen interactions. We investigated the inflammatory response in Anemonia sulcata (Cnidaria: Anthozoa) following injection of substances that varied in type and dimension, and observed clear, strong and specific reactions, especially after injection of Escherichia coli and Vibrio alginolyticus. Moreover, we analyzed enzymatic activity of protease, phosphatase and esterase, showing how the injection of different ba…

Response of Sabella spallanzanii to multiple stressors. The combined effect of infection and copper sulphate

The aim of this work is to study the immune responses of the polychaete Sabella spallanzanii after exposure to copper sulphate, an immunomodulating agent in marine organisms, and the multiple stresses caused by Escherichia coli infection, to validate the species as a model organism in marine-coastal biomonitoring programmes. Polychaetes were housed in laboratory and divided into five experimental groups: 1. Control (no microinjected), 2. filtered seawater + TBS injection (control of point 3), 3. filtered seawater + E. coli injection (control of point 4), 4. CuSO4 + TBS injection (control of point 5), and 5. CuSO4 + E. coli injection. The immune variables, esterase and alkaline phosphatase a…

Preliminare caratterizzazione di un’attività citotossica nel supernatante del lisato degli emociti con granulo rifrangente di Ciona intestinalis

Evolution of Innate Immunity. Components of inflammatory reaction in Ciona intestinalis

The ascidian Styela plicata hemocytes as a potential biomarker of marine pollution: In vitro effects of seawater and organic mercury.

Toxic metals, such as mercury, contribute substantially to anthropogenic pollution in many estuarine environments. Animals living in those environments, particularly invertebrate filter feeders like tunicates, can be used as bioindicators. In an attempt to identify cellular markers for revealing pollution, this study examined in vitro the effects of different concentrations of methyl mercury on Styela plicata hemocytes. The harvested hemocytes from S. plicata that were exposed to the metal had a significant mortality, cellular count and morphometric alterations. These findings provided evidence of MeHg immunotoxic effects on S. plicata, resulting in hemocyte death and morphological changes …

INDAGINI SULLA RISPOSTA IMMUNITARIA DEL PUNTERUOLO ROSSO NEI CONFRONTI DEI NEMATODI ENTOMOPAGENI

Il Punteruolo rosso delle palme (Rhynchophorus ferrugineus Olivier, 1790. Coleoptera Curculionidae), originario dell’Asia meridionale si è velocemente diff uso nell’areale mediterraneo attaccando diversi generi di palme e creando seri problemi al settore ornamentale e vivaistico, oltre che alle aree verdi delle città. Il controllo di questo fi tofago si è rivelato problematico e di diffi cile realizzazione a causa delle sue proprietà invasive. Allo stato attuale non sono stati individuati patogeni effi caci nella lotta biologica, anche se i nematodi entomopatogeni (EPN) sono stati impiegati con discreto successo in Spagna. Generalmente gli EPN sono impiegati contro gli stadi larvali ma poic…

Enhanced expression of a CinTNF gene in the LPS challenged inflammatiry responses of the ascidian Ciona intestinalis

LPS challenge regulates gene expression and tissue localization of a Ciona intestinalis gene through an alternative polyadenylation mechanism.

A subtractive hybridization strategy for the identification of differentially expressed genes was performed between LPS-challenged and naive Ciona intestinalis. This strategy allowed the characterization of two transcripts (Ci8short and Ci8long) generated by the use of two Alternative Polyadenylation sites. The Ci8long transcript contains a protein domain with relevant homology to several components of the Receptor Transporting Protein (RTP) family not present in the Ci8short mRNA. By means of Real Time PCR and Northern Blot, the Ci8short and Ci8long transcripts showed a different pattern of gene expression with the Ci8short mRNA being strongly activated after LPS injection in the pharynx. …

Inflammatory response of Ciona intestinalis involve enhanced D-galactoside-specific lectins and FACIT collagen expression.

Inducible lectins with galectin properties and human IL1alpha epitopes opsonize yeast during the inflammatory response of the ascidian Ciona intestinalis.

Studies on inducible ascidian lectins may shed light on the evolutionary emergence of cytokine functions. Here, we show that the levels of opsonins, with IL1alpha-epitopes, increase in Ciona intestinalis hemolymph as a response to an inflammatory stimulus and, in particular, to intratunic injection of lipopolysaccharide (LPS). The inflammatory agent promptly (within 4 h) enhances Ca(2+)-independent serum hemagglutinating and opsonizing activities, which are both inhibited by D-galactose and D-galactosides (alpha-lactose, N-acetyl-D-lactosamine, thio-digalactoside), suggesting that anti-rabbit erythrocyte lectins with galectin properties are involved as opsonins. Inducible galectin molecules…

Cooperazione cellulare nella reazione citotossica dei celomociti di Paracentrotus lividus

Distinct protoconchs recognised in three of the larger Mediterranean Cerithium species (Caenogastropoda: Cerithiidae)

The gastropod genus Cerithium includes several polymorphic species which are hardly distinguishable using a morphological approach based on teleoconch characters. Here we show that protoconch characters can be reliably used to identify the larger Mediterranean species (Cerithium alucastrum, C. repandum and C. vulgatum), and to assess their intraspecific variability. Based on a large amount of morphological data, we show that a multispiral, strongly sculptured protoconch (traditionally associated with C. vulgatum) is found in C. alucastrum. This species originated in the Pliocene. A multispiral, weakly sculptured protoconch, not observed previously, is reported for C. vulgatum. A paucispiral…

Physiological advantages of dwarfing in surviving extinctions in high-CO2 oceans

Excessive CO2 in the present-day ocean-atmosphere system is causing ocean acidification, and is likely to cause a severe biodiversity decline in the future, mirroring effects in many past mass extinctions. Fossil records demonstrate that organisms surviving such events were often smaller than those before, a phenomenon called the Lilliput effect. Here, we show that two gastropod species adapted to acidified seawater at shallow-water CO2 seeps were smaller than those found in normal pH conditions and had higher mass-specific energy consumption but significantly lower whole-animal metabolic energy demand. These physiological changes allowed the animals to maintain calcification and to partial…