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Davide MALAGOLI
Professore Ordinario
Dipartimento di Scienze della Vita sede ex-Biologia
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Pubblicazioni
2023
- Clodronate Liposome-Mediated Phagocytic Hemocyte Depletion Affects the Regeneration of the Cephalic Tentacle of the Invasive Snail, Pomacea canaliculata
[Articolo su rivista]
Bergamini, Giulia; Sacchi, Sandro; Ferri, Anita; Franchi, Nicola; Montanari, Monica; Ahmad, Mohamad; Losi, Chiara; Nasi, Milena; Cocchi, Marina; Malagoli, Davide
abstract
After amputation, granular hemocytes infiltrate the blastema of regenerating cephalic
tentacles of the freshwater snail Pomacea canaliculata. Here, the circulating phagocytic hemocytes were chemically depleted by injecting the snails with clodronate liposomes, and the effects on the cephalic tentacle regeneration onset and on Pc-Hemocyanin, Pc-transglutaminase (Pc-TG) and Pc-Allograft Inflammatory Factor-1 (Pc-AIF-1) gene expressions were investigated. Flow cytometry analysis demonstrated that clodronate liposomes targeted large circulating hemocytes, resulting in a transient decrease in their number. Corresponding with the phagocyte depletion, tentacle regeneration onset
was halted, and it resumed at the expected pace when clodronate liposome effects were no longer visible. In addition to the regeneration progress, the expressions of Pc-Hemocyanin, Pc-TG, and Pc-AIF-1, which are markers of hemocyte-mediated functions like oxygen transport and immunity, clotting, and inflammation, were modified. After the injection of clodronate liposomes, a specific computer-assisted image analysis protocol still evidenced the presence of granular hemocytes in the tentacle blastema. This is consistent with reports indicating the large and agranular hemocyte population as the most represented among the professional phagocytes of P. canaliculata and with the hypothesis that different hemocyte morphologies could exert diverse biological functions, as it has been observed in other invertebrates.
2023
- The Eco-Immunological Relevance of the Anti-Oxidant Response in Invasive Molluscs
[Articolo su rivista]
Malagoli, Davide; Franchi, Nicola; Sacchi, Sandro
abstract
Reactive oxygen species (ROS) are volatile and short-lived molecules playing important roles in several physiological functions, including immunity and physiological adaptation to unsuit- able environmental conditions. In an eco-immunological view, the energetic costs associated with an advantageous metabolic apparatus able to cope with wide changes in environmental parameters, e.g., temperature range, water salinity or drought, could be further balanced by the advantages that this apparatus may also represent in other situations, e.g., during the immune response. This review provides an overview of molluscs included in the IUCN list of the worst invasive species, highlighting how their relevant capacity to manage ROS production during physiologically challenging situations can also be advantageously employed during the immune response. Current evidence suggests that a relevant capacity to buffer ROS action and their damaging consequences is advantageous in the face of both environmental and immunological challenges, and this may represent a trait for potential invasiveness. This should be considered in order to obtain or update information when investigating the potential of the invasiveness of emerging alien species, and also in view of ongoing climate changes.
2022
- Immune contribution to tentacle regeneration in adult mollusc and cnidarian models
[Abstract in Rivista]
Bergamini, G; Sacchi, S; Ahmad, M; Cocchi, M; Basu, S; Ikmi, A; Malagoli, D
abstract
Histological studies focusing on the early
cephalic tentacle regeneration in P. canaliculata,
have demonstrated that wound closure and
blastema formation took place within 24 h post
amputation (hpa). A Matlab® plugin allowed the
semi-automated identification and quantification of a
phagocytic hemocyte sub-population in the
blastema. Flow cytometry analysis showed that the
injection of the phagocyte-specific drug
Clophosome® (45 µg/g snail) could transiently
remove circulating hemocytes, that recovered the
pre-treatment level within 24 h. Consistently,
histological experiment demonstrated that rare
hemocytes were present in the early regenerating
tentacles of Clophosome®-injected snails
2021
- A New Protocol of Computer-Assisted Image Analysis Highlights the Presence of Hemocytes in the Regenerating Cephalic Tentacles of Adult Pomacea canaliculata
[Articolo su rivista]
Bergamini, Giulia; Ahmad, Mohamad; Cocchi, Marina; Malagoli, Davide
abstract
In humans, injuries and diseases can result in irreversible tissue or organ loss. This wellknown
fact has prompted several basic studies on organisms capable of adult regeneration, such
as amphibians, bony fish, and invertebrates. These studies have provided important biological
information and helped to develop regenerative medicine therapies, but important gaps concerning
the regulation of tissue and organ regeneration remain to be elucidated. To this aim, new models
for studying regenerative biology could prove helpful. Here, the description of the cephalic tentacle
regeneration in the adult of the freshwater snail Pomacea canaliculata is presented. In this invasive
mollusk, the whole tentacle is reconstructed within 3 months. Regenerating epithelial, connective,
muscular and neural components are already recognizable 72 h post-amputation (hpa). Only in
the early phases of regeneration, several hemocytes are retrieved in the forming blastema. In
view of quantifying the hemocytes retrieved in regenerating organs, granular hemocytes present
in the tentacle blastema at 12 hpa were counted, with a new and specific computer-assisted image
analysis protocol. Since it can be applied in absence of specific cell markers and after a common
hematoxylin-eosin staining, this protocol could prove helpful to evidence and count the hemocytes
interspersed among regenerating tissues, helping to unveil the role of immune-related cells in sensory
organ regeneration.
2021
- Expression pattern of perilipins in human brain during aging and in Alzheimer's disease
[Articolo su rivista]
Conte, Maria; Medici, Valentina; Malagoli, Davide; Chiariello, Antonio; Cirrincione, Alice; Davin, Annalisa; Chikhladze, Maia; Vasuri, Francesco; Legname, Giuseppe; Ferrer, Isidre; Vanni, Silvia; Marcon, Gabriella; Poloni, Tino Emanuele; Guaita, Antonio; Franceschi, Claudio; Salvioli, Stefano
abstract
Perilipins are conserved proteins that decorate intracellular lipid droplets and are essential for lipid metabolism. To date, there is limited knowledge on their expression in human brain or their involvement in brain aging and neurodegeneration. The aim of this study was to characterise the expression levels of perilipins (Plin1-Plin5) in different cerebral areas from subjects of different age, with or without signs of neurodegeneration.
2021
- Pomacea canaliculata ampullar proteome: A nematode-based bio-pesticide induces changes in metabolic and stress-related pathways
[Articolo su rivista]
Boraldi, F.; Lofaro, F. D.; Bergamini, G.; Ferrari, A.; Malagoli, D.
abstract
Pomacea canaliculata is a freshwater gastropod known for being both a highly invasive species and one of the possible intermediate hosts of the mammalian parasite Angiostrongylus cantonensis. With the aim of providing new information concerning P. canaliculata biology and adaptability, the first proteome of the ampulla, i.e., a small organ associated with the circulatory system and known as a reservoir of nitrogen-containing compounds, was obtained. The ampullar proteome was derived from ampullae of control snails or after exposure to a nematode-based molluscicide, known for killing snails in a dose-and temperature-dependent fashion. Proteome analysis revealed that the composition of connective ampulla walls, cell metabolism and oxidative stress response were affected by the biopesticide. Ultrastructural investigations have highlighted the presence of rhogocytes within the ampullar walls, as it has been reported for other organs containing nitrogen storage tissue. Collected data suggested that the ampulla may belong to a network of organs involved in controlling and facing oxidative stress in different situations. The response against the nematode-based molluscicide recalled the response set up during early arousal after aestivation and hibernation, thus encouraging the hypothesis that metabolic pathways and antioxidant defences promoting amphibiousness could also prove useful in facing other challenges stimulating an oxidative stress response, e.g., immune challenges or biocide exposure. Targeting the oxidative stress resistance of P. canaliculata may prove helpful for increasing its susceptibility to bio-pesticides and may help the sustainable control of this pest’s diffusion.
2020
- 20-OH-ecdysone and extracellular ATP are not pro-autophagic factors for the lepidopteran fat bodycell line, IPLB-LdFB
[Abstract in Atti di Convegno]
Ferrari, A; Fiorino, R; Montanari, M; Accorsi, A; Simonini, R; de Eguileor, M; Malagoli, D
abstract
2020
- Hemocyte depletion as a tool for studying immune cell dynamics and contribution to fundamental biological processes in the freshwater snail Pomacea canaliculata
[Abstract in Atti di Convegno]
Bergamini, Giulia; Accorsi, Alice; Malagoli, Davide
abstract
2020
- Organ-specific accumulation of Superparamagnetic Iron Oxide Nanoparticles (SPIONs) in the apple snail Pomacea canaliculata
[Poster]
Fiorino, Roberta; Bergamini, Giulia; Ahmad, Mohamad; Cocchi, Marina; Prina-Mello, Adriele; Malagoli, Davide
abstract
2020
- The apple snail Pomacea canaliculata: a new and alternative animal model for testing innovative nanomedicines
[Relazione in Atti di Convegno]
Fiorino, Roberta; Bergamini, Giulia; Tosi, Giovanni; Prina-Mello, Adriele; Malagoli, Davide
abstract
2020
- The immune response of the invasive golden apple snail to a nematode‐based molluscicide involves different organs
[Articolo su rivista]
Montanari, Alice; Bergamini, Giulia; Ferrari, Agnese; Ferri, Anita; Nasi, Milena; Simonini, Roberto; Malagoli, Davide
abstract
Simple SummarySustainable solutions to the spreading of invasive species are difficult to find due to the absence of biological information about basic immune mechanisms of the target pests. Here, we present evidence of the effects of a commercially available roundworm, Phasmarhabditis hermaphrodita, against the invasive apple snail Pomacea canaliculata. The effects are principally evaluated in terms of snail survival and immune activation. Via molecular and microscopy-based approaches, we demonstrate that dosage and temperature are critical in determining the effects of the roundworm, and that the apple snail response to this immune challenge involves different organs. To our knowledge, these findings are the first demonstration that a P. hermaphrodita-based molluscicide can effectively kill P. canaliculata and that the snail can mount a multi-organ response against this pathogenic roundworm.The spreading of alien and invasive species poses new challenges for the ecosystem services, the sustainable production of food, and human well-being. Unveiling and targeting the immune system of invasive species can prove helpful for basic and applied research. Here, we present evidence that a nematode (Phasmarhabditis hermaphrodita)-based molluscicide exerts dose-dependent lethal effects on the golden apple snail, Pomacea canaliculata. When used at 1.7 g/L, this biopesticide kills about 30% of snails within one week and promotes a change in the expression of Pc-bpi, an orthologue of mammalian bactericidal/permeability increasing protein (BPI). Changes in Pc-bpi expression, as monitored by quantitative PCR (qPCR), occurred in two immune-related organs, namely the anterior kidney and the gills, after exposure at 18 and 25 degrees C, respectively. Histological analyses revealed the presence of the nematode in the snail anterior kidney and the gills at both 18 and 25 degrees C. The mantle and the central nervous system had a stable Pc-bpi expression and seemed not affected by the nematodes. Fluorescence in situ hybridization (FISH) experiments demonstrated the expression of Pc-bpi in circulating hemocytes, nurturing the possibility that increased Pc-bpi expression in the anterior kidney and gills may be due to the hemocytes patrolling the organs. While suggesting that P. hermaphrodita-based biopesticides enable the sustainable control of P. canaliculata spread, our experiments also unveiled an organ-specific and temperature-dependent response in the snails exposed to the nematodes. Overall, our data indicate that, after exposure to a pathogen, the snail P. canaliculata can mount a complex, multi-organ innate immune response.
2020
- The potential immune alterations in insect pests and pollinators after insecticide
exposure in agroecosystem
[Articolo su rivista]
Zibaee, A; Malagoli, D
abstract
Agroecosystems are the habitat of pests and beneficial insects from different orders, which are
exposed to agro-practices, especially treatments with chemicals. Insecticides are a wide group of
chemicals used in agroecosystems that affect insect ecology and physiology in different ways. Among
physiological components affected by insecticides, the immune system (IS) is an important one,
enabling insects to resist against invading microorganisms and parasitoids thanks to the action of
hemocytes and humoral components. So the determination of any immune alterations should be
considered as a critical issue in insecticide application within agroecosystems. Insecticides of synthetic
or natural origin, e.g. insect growth regulators (IGRs) and botanicals, are frequently cytotoxic and alter
hemocyte morphology and number, impairing cellular-based immune responses in addition to humeral
responses. Exposure of pollinators to neurotoxin insecticides like neonicotinoids may inhibit the
immune-related transcription factor, NF- B, with a negative impact on the expression of antimicrobial
peptides, melanization and clotting. In contrast, some IGRs may have enhancing effects on hemocyte
spreading mainly plasmatocytes and cellular-based immune responses. Chemical insecticides have
several impacts on the physiology of insects in which immune modulation is one of the most important
cases because any alteration may alter their ability to respond toward invading pathogens and directly
their survival. This is more severe once pollinators are in contact with chemicals because of the
presence of several pathogenic agents that directly influence their performance.
2019
- Effects of a nematode-based molluscicide on survival and antimicrobial peptide expression in Pomacea canaliculata
[Abstract in Atti di Convegno]
Montanari, A; Accorsi, A; Nasi, M; Malagoli, D
abstract
2019
- HEMOCYTE RESERVOIRS IN THE MOLLUSC Pomacea canaliculata: EVIDENCE FROM MORHPOLOGICAL AND IMAGE-BASED FLOW CYTOMETRY ANALYSES
[Abstract in Atti di Convegno]
Montanari, Alice; Accorsi, Alice; Box, Andrew; Ferrari, Agnese; Malagoli, Davide
abstract
2019
- In vivo testing of nanomedicines: Seeking New Animal models for Investigating Life
[Relazione in Atti di Convegno]
Malagoli, Davide
abstract
2019
- Toward the Molecular Deciphering of Pomacea canaliculata Immunity: First Proteomic Analysis of Circulating Hemocytes
[Articolo su rivista]
Boraldi, F.; Lofaro, FRANCESCO DEMETRIO; Accorsi, A.; Ross, E.; Malagoli, D.
abstract
Pomacea canaliculata is a freshwater snail with interesting biological features that include invasiveness, human parasite hosting, and adult regeneration. Its immune system may represent the target for strategies aimed at controlling the spread of the snail population and its hosting of the human parasite Angiostrongylus cantonensis. Moreover, immune functions likely have a role in the snail's ability to wound heal and regenerate. Despite its importance in multiple processes, very little is known about the molecular basis of P. canaliculata immunity. Aiming to contribute to filling this gap, the ultrastructure of circulating hemocytes in healthy snails is studied and the first proteomic analysis of these cells is performed, evidencing 83 unique proteins, 96% of which have identifiable homologs in other species. Fifteen proteins are retrieved as potentially involved in immune-related signaling pathways, such as hemocyanin, C1q-like protein, and HSP90 together with cytoskeleton and cytoskeleton-related proteins involved in cell motility and membrane dynamics. This first proteome study on non-stimulated hemocytes provides a valid reference for future investigations on the molecular changes under stressful circumstances, like pathogen exposure, wounding, or environmental changes.
2018
- COMBINED METHODS TO INVESTIGATE AT HIGH RESOLUTION THE COMPLEXITY OF HEMATOPOIESIS IN A MOLLUSCAN RESEARCH ORGANISM
[Abstract in Atti di Convegno]
Accorsi, Alice; Box, Andrew; Ross, Eric; Malagoli, Davide
abstract
2018
- Going beyond a static picture: the apple snail Pomacea canaliculata can tell us the life history of molluscan hemocytes
[Articolo su rivista]
Malagoli, D
abstract
More than 40 years of studies on molluscan immunity have revealed a complex and dynamic immune system endowed with multifunctional circulating cells, i.e., hemocytes that are regulated by diverse signaling molecules. However, very little is known about the dynamic processes that drive hemocyte proliferation, differentiation, maturation, and senescence. Evidence reported here highlights how the apple snail Pomacea canaliculata is an extremely promising research organism that will provide answers to the numerous questions regarding the life-history of molluscan and lophotrochozoan hemocytes.
2018
- Real time PCR and image-based cell profiling analyses evidence correlation between hematopoietic and ampulla cell populations in the gastropod Pomacea canaliculata
[Abstract in Atti di Convegno]
Accorsi, A; Montanari, A; Nasi, M; Pecorini, S; Box, A; Peuß, R; Malagoli, D
abstract
2017
- A prokineticin-like protein responds to immune challenges in the gastropod pest Pomacea canaliculata
[Articolo su rivista]
Accorsi, Alice; Benatti, Stefania; Ross, Eric; Nasi, Milena; Malagoli, Davide
abstract
The golden apple snail Pomacea canaliculata is an invasive pest originating from South America. It has already been found in Asia, the southern United States and more recently in the EU. Aiming to target the immune system of the snail as a way to control its spreading, we have developed organ-specific transcriptomes and looked for molecules controlling replication and differentiation of snail hemocytes. The prokineticin domain-containing protein Astakine 1 is the only cytokine known thus far capable of regulating invertebrate hematopoiesis, and we analyzed the transcriptomes looking for molecules containing a prokineticin domain. We have identified a prokineticin-like protein (PlP), that we called Pc-plp and we analyzed by real-time PCR (qPCR) its expression. In control snails, highest levels of Pc-plp were detected in the digestive gland, the ampulla (i.e., a hemocyte reservoir) and the pericardial fluid (i.e., the hematopoietic district). We tested Pc-plp expression after triggering hematopoiesis via multiple hemolymph withdrawals, or during bacterial challenge through LPS injection. In both cases a reduction of Pc-plp mRNA was observed. The multiple hemolymph withdrawals caused a significant decrease of Pc-plp mRNA in pericardial fluid and circulating hemocytes, while the LPS injection promoted the Pc-plp mRNA drop in anterior kidney, mantle and gills, organs that may act as immune barrier in molluscs. Our data indicate an important role for prokineticin domain-containing proteins as immunomodulators also in gastropods and their dynamic expression may serve as a biosensor to gauge the effectiveness of immunological interventions aimed at curtailing the spreading of the gastropod pest P. canaliculata.
2017
- Circulating phagocytes: The ancient and conserved interface between immune and neuroendocrine function
[Articolo su rivista]
Malagoli, Davide; Mandrioli, Mauro; Tascedda, Fabio; Ottaviani, Enzo
abstract
Immune and neuroendocrine functions display significant overlap in highly divergent and evolutionarily distant models such as molluscs, crustaceans, insects and mammals. Fundamental players in this crosstalk are professional phagocytes: macrophages in vertebrates and immunocytes in invertebrates. Although they have different developmental origins, macrophages and immunocytes possess comparable functions and differentiate under the control of evolutionarily conserved transcription factors. Macrophages and immunocytes share their pools of receptors, signalling molecules and pathways with neural cells and the neuro-endocrine system. In crustaceans, adult transdifferentiation of circulating haemocytes into neural cells has been documented recently. In light of developmental, molecular and functional evidence, we propose that the immune-neuroendocrine role of circulating phagocytes pre-dates the split of protostomian and deuterostomian superphyla and has been conserved during the evolution of the main groups of metazoans.
2017
- Cross-talk among immune and neuroendocrine systems in molluscs and other invertebrate models
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
2016
- . Pc-astakine mRNA levels decrease after both LPS injection and hemolymph withdrawals in the gastropod pest Pomacea canaliculata.
[Abstract in Rivista]
Accorsi, Alice; Gotting, Karen; Ross, Eric; Nasi, Milena; Ottaviani, Enzo; Sánchez Alvarado, Alejandro; Malagoli, Davide
abstract
The freshwater gastropod Pomacea canaliculata has recently been included among emerging pests in the EU. Hence, its immune system may be a promising target for pest control strategies. By analyzing a set of P. canaliculata transcriptomes, we have discovered an Astakine-1-like molecule (Pc-Astakine) in this organism. Astakine was first isolated as a hematopoietic cytokine in the crayfish Pacifastacus leniusculus (Pl-Astakine-1), and later found in arthropods and the bivalve Crassostrea gigas. Bioinformatic analyses show that Pc-Astakine is a 121 aa protein with a conserved distribution pattern of cysteines retrievable in both Pl-Astakine-1 and the vertebrate Prokineticin protein family. Pc-astakine is expressed at low levels in circulating hemocytes, but markedly in hemocyte reservoirs (e.g., ampulla) and hematopoietic tissue (pericardial fluid) suggesting a conserved role in hematopoiesis. We tested this potential role of Pc-Astakine in two ways. First, we measured RNA levels of this gene in immune-related tissues 24 h after injection of 50 μg of Escherichia coli-derived lipopolysaccharide (055:B5) (LPS). We observed a sharp decrease of Pc-astakine mRNA in all the analyzed organs, with Pc-astakine mRNA levels almost undetectable in anterior kidney (a hemocyte reservoir) when compared to sham-injected controls. Second, we defined the profile of Pc-astakine expression after a series of 4 hemolymph withdrawals performed within 72 h. As with the LPS-treatment, a general decrease of mRNA levels was observed, with the largest decrease observed in hematopoietic tissue and circulating hemocytes (reduced to 20% of the control). Altogether, we hypothesize that the decrease of Pc-astakine mRNA after both LPS injection and hemolymph withdrawal may be due to an increased rate of translation, rather than a reduction of gene expression. This is in part supported by similar observations in the oyster C. gigas in which massive translation of Cg-astakine is observed as a consequence of environment stress, including immune stimuli, and in P. leniusculus in which increased levels of Astakine in the plasma of LPS-injected animals have been related to an increased translational rate.
To our knowledge, this is the first report of an astakine-like molecule in gastropods and our data indicate a potential involvement of Pc-Astakine in both the immune response and hematopoiesis of the molluscan pest P. canaliculata.
2016
- ACTH in invertebrates: A molecule for all seasons
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
In vertebrate and invertebrate models, adrenocorticotropic hormone (ACTH) belongs to the melanocortin group of related peptides, which share a common precursor, pro-opiomelanocortin (POMC). Functional experiments indicate that in invertebrates, ACTH plays a major role in several biological functions. ACTH, whose effects have been conserved during evolution more than its amino acidic sequence, is, directly or indirectly, able to contrast agents that perturb a body’s homeostasis. Here we review evidence highlighting the involvement of ACTH and ACTH-like molecules in the response of invertebrate models versus immune, environmental and parasitic challenges.
2016
- Cell Death Pathways in an Unconventional Invertebrate Model
[Capitolo/Saggio]
Ottaviani, Enzo; Malagoli, Davide
abstract
Programmed cell death (including apoptosis and autophagy-mediated cell death) is a fundamental feature of many important biological processes devoted to the maintenance of the body's homeostasis. Numerous research studies on programmed cell death included well-established invertebrate models like Drosophila melanogaster and Caenorhabditis elegans, and their contribution to the present knowledge on cell death was decisive. Despite this, several questions remain, especially in the relationships existing between the diverse modalities of cell death, asking for further experiments in different models. Here, we describe the prodeath effects and the pathways activated by the mitochondria-targeting drugs 2-deoxy-. d-ribose, sodium nitroprusside, or oligomycin A in the IPLB-LdFB cell line derived from the unconventional insect model Lymantria dispar.
2016
- Hematopoiesis and hemocytes in pancrustacean and molluscan models
[Capitolo/Saggio]
Smith, V. J.; Accorsi, A.; Malagoli, Davide
abstract
The hematopoietic districts and their organization are here described in detail for some of the principal pancrustacean and molluscan models. Pancrustaceans present an overall similar organization of hematopoietic organs, while among mollusks the hematopoietic organs and tissues are more diversified. The morphologies, functions, and main lineages of circulating hemocytes are recapitulated, and the common traits between different taxa are evidenced
2016
- Pomacea canaliculata: development and regeneration of complex eyes.
[Abstract in Atti di Convegno]
Accorsi, Alice; Ross, Eric; Malagoli, Davide; Sánchez Alvarado, Alejandro
abstract
A systematic molecular and genetic dissection of complex camera-type eye regeneration has been hampered by the
paucity of model systems in which to carry out these studies. The freshwater snail Pomacea canaliculata has a number of
biological attributes that lend this organism ideally suited for studies in development and regeneration. Chiefly among
them are ease of culture and reproduction in captivity with abundant and year-round offspring production, direct
development and the ability to regenerate complex organs, such as tentacles and eyes after complete amputation. During
embryogenesis, the eyes are developed relatively early; the retina, the lens and the general structure of the eyes are already
well formed and similar to the adult eyes prior to hatching. Once amputated, the adult complex camera-type eyes,
comprised of cornea, lens, retina and optic nerve, can fully regenerate in a relative short time (1 month) and the de novo
regenerating eye shows a complete reconstruction of all the mentioned layers. To advance our studies, we have generated
a high-dimensional transcriptome at high temporal resolution of the regenerating P. canaliculata eye. Equivalent work has
been done during embryogenesis, providing us with an opportunity to compare embryonic versus regeneration eye
ontogeny. Our aim is to determine whether embryonic morphogenetic programs and pathways are reactivated in adult eye
regeneration and to identify the stimuli that trigger the reactivation of cell proliferation and morphogenesis of a new
complex camera-type eye in adult P. canaliculata.
2016
- Preface [The evolution of the immune system]
[Breve Introduzione]
Malagoli, Davide
abstract
Brief Preface in quality of Editor of the Book
2016
- The evolution of the immune system: conservation and diversification
[Curatela]
Malagoli, Davide
abstract
The Evolution of the Immune System: Conservation and Diversification is the first book of its kind that prompts a new perspective when describing and considering the evolution of the immune system. Its unique approach summarizes, updates, and provides new insights on the different immune receptors, soluble factors, and immune cell effectors. Helps the reader gain a modern idea of the evolution of the immune systems in pluricellular organisms Provides a complete overview of the most studied and hot topics in comparative and evolutionary immunology Reflects the organisation of the immune system (cell-based, humoral [innate], humoral [adaptive]) without introducing further and misleading levels of organization Brings concepts and ideas on the evolution of the immune system to a wide readership.
2015
- Autophagic cell death in the insect cell line IPLB-LdFB: evidence in absence of known mediators
[Abstract in Rivista]
Accorsi, Alice; Bellelli, A.; Ottaviani, Enzo; Malagoli, Davide
abstract
Autophagic cell death in the insect cell line IPLB-LdFB
2015
- Hematopoiesis and hemocyte reservoirs control the number of circulating hemocytes in the apple snail Pomacea canaliculata (Mollusca, Gastropoda)
[Abstract in Atti di Convegno]
Accorsi, Alice; Ottaviani, Enzo; Malagoli, Davide
abstract
Pomacea canaliculata is an invasive freshwater snail native to South America and introduced in Southeast Asia, where it severely damages rice seedlings and acts as intermediate host of the parasite Angiostrongylus cantonensis. After its retrieval in Spain, the European Union Parliament banned P. canaliculata import and diffusion. Information about the immune system of P. canaliculata is needed to implement the control measures against its diffusion.
The principal cellular players of molluscan immune system are the circulating hemocytes. P. canaliculata hemocytes consist of blast-like cells (20%), hyalinocytes (72%) and granulocytes (8%). All these cells adhere to glass surface, but only hyalinocytes and granulocytes phagocytize heat-inactivated bacteria. Four hemolymph withdrawals in 72 h did not change the hemocyte total number but blast-like cells slightly decreased. The p(Ser10)-H3 mitotic marker was evidenced in cells along the external side of the vessels entering the heart. These observations were indicative of an active hematopoietic tissue, localized into the pericardial cavity. The repeated withdrawals had no effects on hemocyte renewal rate, thus one or more hemocyte reservoirs may intervene in buffering hemocyte lost in P. canaliculata. Light microscopy observations suggested that the ampulla acts as hemocyte reservoir.
These results demonstrate that adult P. canaliculata possess an active hematopoietic district like Biomphalaria glabrata and other gastropods, whose amebocyte-producing organ (APO) consists of clusters of cells positioned between the pericardial and the posterior mantle epithelia. Further, hematopoiesis role in hemocyte renewal is supported by the ampulla, that seems to be a hemocyte reservoir in P. canaliculata.
2015
- Molluscs as models for translational medicine
[Articolo su rivista]
Tascedda, Fabio; Malagoli, Davide; Accorsi, Alice; Rigillo, Giovanna; Blom, Johanna Maria Catharina; Ottaviani, Enzo
abstract
This paper describes the advantages of adopting a molluscan model for studying the biological basis of some central nervous system pathologies affecting humans. In particular, we will focus on the freshwater snail Lymnaea stagnalis, which is already the subject of electrophysiological studies related to learning and memory, as well as ecotoxicological studies. The genome of L. stagnalis has been sequenced and annotated but the gene characterization has not yet been performed. We consider the characterization of the gene networks that play crucial roles in development and functioning of the central nervous system in L. stagnalis, an important scientific development that comparative biologists should pursue. This important effort would add a new experimental model to the limited number of invertebrates already used in studies of translational medicine, the discipline that seeks to improve human health by taking advantage of knowledge collected at the molecular and cellular levels in non-human organisms.
2015
- The multifaceted activity of insect caspases
[Articolo su rivista]
Accorsi, Alice; Zibaee, A; Malagoli, Davide
abstract
Caspases are frequently considered synonymous with apoptotic cell death. Increasing evidence demonstrates that these proteases may exert their activities in non-apoptotic functions. The non-apoptotic roles of caspases may include developmentally regulated autophagy during insect metamorphosis, as well as neuroblast self-renewal and the immune response. Here, we summarize the established knowledge and the recent advances in the multiple roles of insect caspases to highlight their relevance for physiological processes and survival.
2014
- Autophagy researchers
[Scheda bibliografica]
Carra, Serena; Malagoli, Davide; Ney, Pa; Steffan, Js
abstract
Brief description of the various PI works and interest, especially centered on the autophagy field
2014
- Eco-immunology-Evolutive Aspects and Future Perspectives
[Monografia/Trattato scientifico]
Malagoli, Davide; Ottaviani, Enzo
abstract
THE PRODUCT IS A BOOK
2014
- Effects of repeated hemolymph withdrawals on the hemocyte populations and hematopoiesis in Pomacea canaliculata
[Articolo su rivista]
Accorsi, Alice; Ottaviani, Enzo; Malagoli, Davide
abstract
Pomacea canaliculata is a freshwater gastropod considered an invasive pest by several European, North
American and Asiatic countries. This snail presents a considerable resistance to pollutants and may
successfully face stressful events. Thanks to the unusual possibility to perform several hemolymph
collections without affecting its survival, P. canaliculata is a good model to study the hematopoietic
process and the hemocyte turnover in molluscs. Here we have analyzed the effects of repeated hemolymph
withdrawals on circulating hemocyte populations and pericardial organs, i.e., the heart, the main
vessels entering and leaving the heart and the ampulla, of P. canaliculata. Our experiments revealed that
the circulating hemocyte populations were maintained constant after 3 collections performed in 48 h.
The tissue organization of the heart and the vessels remained unaltered, whereas the ampulla buffered
the effects of hemolymph collections acting as hemocyte reservoir, and its original organization was
progressively lost by the repeated hemolymph withdrawals. The hematopoietic tissue of P. canaliculata
was evidenced here for the first time. It is positioned within the pericardial cavity, in correspondence of
the principle veins. Mitoses within the hematopoietic tissue were not influenced by hemolymph collections,
and circulating hemocytes never presented mitotic activity.
2014
- Functional amyloid formation in LPS activated cells from invertebrates to vertebrates
[Articolo su rivista]
Grimaldi, A.; Tettamanti, G.; Girardello, R.; Pulze, L.; Valvassori, R.; Malagoli, D.; Ottaviani, E.; de Eguileor, M.
abstract
LPS stimulation provokes serious cellular stress with an increase of cytoplasmic reactive oxygen species (ROS). We have investigated, among the different cellular defenses, amyloidogenesis as common physiological response to attenuate oxidative stress. Optical and electron microscopic observations of the following LPS activated cell lines [insect (larval hemocytes, IPLB-LdFB and Drosophila Schneider’s S2 cells); mouse (NIH3T3 embryonic fibroblasts); Human (Human Umbilical Vein Endothelial Cells (HUVEC), neutrophils, and mesenchymal stem cells] reveal that, all are characterized by irregular profiles, cytoplasmic empty vacuoles or by cisternae containing fibrillar material. The compartmentalized fibrillar material shows staining properties typical of amyloid fibrils. LPS activation leads to ROS generation, resulting in pH acidification. Stimulated cells show pink cytoplasm in May-Grünwald Giemsa differential staining, giving a gross indication of a lower intracellular pH. Moreover the activation of amyloidogenesis is also linked with an extensive production of ACTH and α-MSH in all cultured cell types. We suggest that amyloidogenesis is a common, physiological cellular response to weak ROS, starting when other anti-stress cellular systems failed to restore homeostasis. The morphological evidence and/or functional characterization of synthesized amyloid fibrils could be an early indicator of oxidative stress that may lead to a general inflammatory process.
2014
- Hematopoiesis and hemocyte replacement after repeated hemolymph withdrawals in Pomacea canaliculata (Mollusca, Gastropoda)
[Abstract in Atti di Convegno]
Accorsi, Alice; Malagoli, Davide; Ottaviani, Enzo
abstract
Hematopoiesis and hemocyte replacement after repeated hemolymph withdrawals in Pomacea canaliculata (Mollusca, Gastropoda)
2014
- Immune response of Chilo suppressalis Walker (Lepidoptera: Crambidae) larvae to different entomopathogenic fungi
[Articolo su rivista]
A., Zibaee; Malagoli, Davide
abstract
The current study reports mortality and effects on cellular immune response of
several entomopathogenic fungi including isoleates BB1, BB2 and BB3 of Beauveria
bassiana, Metarhizium anisopliae, Isaria fumosoroseus and Lecanicilium lecanii against
larvae of Chilo suppressalis. Prohemocytes, granulocytes, plasmatocytes and
oenocytoids were identified as the main circulating hemocytes in the hemolymph
of larvae using Giemsa staining solution. Entomopathogenic fungi caused
differential mortality on larvae: BB1, BB3, M. anisopliae lead to the highest mortality
16 on larvae and L. lecanii caused the lowest mortality. The highest numbers of total
hemocytes were observed 3h post-injection of B. bassiana isolates and 6h for the other
treatments. The highest numbers of plasmatocytes were observed 3h post-injection of
BB1 and Tween 80, whereas BB2, BB3, M. anisopliae, I. fumosoroseus and L. lecani caused plasmatocyte increase 6h post-injection. Similar results were obtained in case
of granulocytes but only Tween 80 showed the highest number of hemocytes 3h post-
injection. The highest numbers of nodules were found at various time intervals after
injection of fungal isolates and latex bead. The highest activities of phenoloxidase
were observed 12h post-injection by BbB1, BbB3, M. anisopliae and latex bead; 3–6h
post-injection by BbB2, 6h post-injection by I. fumosoroseus and 3–6h post-injection by
L. lecanii. Our data demonstrate the possibility of utilizing different fungal extracts in
the field to help reduce the risk of resistance evolution in C. suppressalis and
encourage experimentations aimed to increase the number of biological control agent
for insect pests such as the striped rice stem borer C. suppressalis.
2014
- Immune-neuroendocrine integration and its evolution.
[Capitolo/Saggio]
Malagoli, Davide; Ottaviani, Enzo
abstract
Immune-neuroendocrine integration and its evolution.
2014
- Lectine, citochine e molecole altamente variabili negli invertebrati
[Capitolo/Saggio]
Malagoli, Davide
abstract
Il presente capitolo affronta un tema di grande attualità per l’immunologia comparata, e prende spunto da scoperte avvenute essenzialmente nell’ultimo decennio. Il concetto di molecole altamente variabili ed estremamente specifiche nell’ambito del sistema immunitario è stato per lungo tempo ritenuto sinonimo di anticorpi di Vertebrato. Oggi è invece dimostrato che i principali e più diffusi taxa di invertebrati quali Molluschi, Insetti, Crostacei, Echinodermi e Tunicati sono in grado di produrre molecole altamente variabili che caratterizzano in modo specifico ogni singolo individuo. Inoltre, il pool di molecole variabili prodotto da un individuo può variare in funzione dell’agente patogeno incontrato, indicando perciò la possibilità di plasmare la risposta immunitaria sulla base della storia individuale del singolo animale e della specifica contingenza che si trova ad affrontare. Il funzionamento del sistema immunitario di questi animali si basa su molecole ancora in larga parte non caratterizzate della classe delle lectine e delle citochine, che unitamente alle molecole variabili permettono di modulare la risposta in funzione dell’intensità e della tipologia dello stimolo. Pur non essendoci evidenze certe di memoria immunitaria, anche al di fuori dei Vertebrati è invece dimostrata la capacità del sistema immunitario di adeguarsi in modo specifico all’ambiente circostante, e pertanto le parole chiave una volta assegnate al sistema immunitario dei Vertebrati, ossia specificità (specificty) ed adattabilità (adaptivity) sono da considerarsi patrimonio comune di tutti i Bilateri.
2014
- Mouse models as paradigms of human diseases.
[Capitolo/Saggio]
Demetrius, L. A.; Malagoli, Davide
abstract
Mouse models as paradigms of human diseases.
2014
- Preface
[Breve Introduzione]
Malagoli, D.; Ottaviani, E.
abstract
2014
- The ampulla is a hemocyte reservoir but not an hematopoietic organ in the freshwater snail Pomacea canaliculata.
[Abstract in Rivista]
Malagoli, Davide; Accorsi, Alice; Ottaviani, Enzo
abstract
The ampulla is a hemocyte reservoir but not an hematopoietic organ in the freshwater snail Pomacea canaliculata.
2013
- Amyloidogenesis as protective response in invertebrates and vertebrates.
[Abstract in Atti di Convegno]
Eguileor De Girardello, R. M.; Grimaldi, A.; Malagoli, Davide; Pulze, L.; Tettamanti, G.; Ottaviani, Enzo; Valvassori, R.
abstract
Amyloidogenesis as protective response in invertebrates and vertebrates.
2013
- Amyloidogenesis as stress response: winner or loser process?
[Abstract in Rivista]
De Eguileor, M.; Grimaldi, A.; Girardello, R; Malagoli, Davide; Pulze, L.; Tettamanti, G.; Ottaviani, Enzo; Valvassori, R.
abstract
Amyloidogenesis as stress response
2013
- Comparative analysis of circulating hemocytes of the freshwater snail Pomacea canaliculata
[Articolo su rivista]
Accorsi, Alice; L., Bucci; M., de Eguileor; Ottaviani, Enzo; Malagoli, Davide
abstract
Molluscs are invertebrates of great relevance for economy, environment and public health. The numerous
studies on molluscan immunity and physiology registered an impressive variability of circulating hemocytes.
This study is focused on the first characterization of the circulating hemocytes of the freshwater
gastropod Pomacea canaliculata, a model for several eco-toxicological and parasitological researches.
Flow cytometry analysis identified two populations of hemocytes on the basis of differences in size and
internal organization. The first population contains small and agranular cells. The second one displays
major size and a more articulated internal organization. Light microscopy evidenced two principal
morphologies, categorized as Group I (small) and II (large) hemocytes. Group I hemocytes present the
characteristics of blast-like cells, with an agranular and basophilic cytoplasm. Group I hemocytes can
adhere onto a glass surface but seem unable to phagocytize heat-inactivated Escherichia coli. The majority
of Group II hemocytes displays an agranular cytoplasm, while a minority presents numerous granules.
Agranular cytoplasm may be basophilic or acidophilic. Granules are positive to neutral red staining and
therefore acidic. Independently from their morphology, Group II hemocytes are able to adhere and to
engulf heat-inactivated E. coli. Transmission electron microscopy analysis clearly distinguished between
agranular and granular hemocytes and highlighted the electron dense content of the granules. After
hemolymph collection, time-course analysis indicated that the Group II hemocytes are subjected to an
evident dynamism with changes in the percentage of agranular and granular hemocytes. The ability of
circulating hemocytes to quickly modify their morphology and stainability suggests that P. canaliculata is
endowed with highly dynamic hemocyte populations able to cope with rapid environmental changes as well as fast growing pathogens.
2013
- Drosophila Helical factor (Hf) and helical cytokine evolution: from in silico approaches to functional evidence.
[Abstract in Atti di Convegno]
Malagoli, Davide
abstract
Summa of present day knowledge about the helical cytokine Helical factor of Dorsophila melanogaster
2013
- Epigenetic modification in neurons of the mollusc Pomacea canaliculata after immune challenge
[Articolo su rivista]
Ottaviani, Enzo; Accorsi, Alice; Rigillo, Giovanna; Malagoli, Davide; Joan M. C., Blom; Tascedda, Fabio
abstract
In human and rodents, the transcriptional response of neurons to stress is related to epigenetic modifications of both DNA and histone proteins. To assess the suitability of simple invertebrate models in studying the basic mechanisms of stress-related epigenetic modifications, we analyzed epigenetic modifications in neurons of the freshwater snail Pomacea canaliculata after the injection of Escherichia coli-derived lipopolysaccharide (LPS). The phospho-acetylation of histone H3, together with the induction of stress-related factors, c-Fos and HSP70, were evaluated in large and small neurons of the pedal ganglia of sham- and LPS-injected snails. Immunocytochemical investigations showed that after LPS injection, the immunopositivity towards phospho (Ser10)-acetyl (Lys14)-histone H3 and c-Fos increases in the nuclei of small gangliar neurons. Western blot analysis confirmed a significant increase of phospho (Ser10)-acetyl (Lys14)-histone H3 in nuclear extracts from 2h LPS-injected animals. c-Fos protein levels were significantly augmented 6h after LPS injection. Immunocytochemistry and western blot indicated that no changes occurred in HSP70 distribution and protein levels. To our knowledge this is the first demonstration of epigenetic changes in molluscan neurons after an immune challenge and indicate the gastropod P. canaliculata as a suitable model for evolutionary and translational studies on stress-related epigenetic modifications.
2013
- Neuropeptide S stimulates human monocyte chemotaxis via NPS receptor activation.
[Articolo su rivista]
Filaferro, Monica; Novi, Chiara; Ruggieri, Valentina; Genedani, Susanna; Alboni, Silvia; Malagoli, Davide; Caló, G; Guerrini, R; Vitale, Giovanni
abstract
Neuropeptide S (NPS) produces several biological actions by activating a formerly orphan GPCR, now named NPS receptor (NPSR). It has been previously demonstrated that NPS stimulates murine leukocyte chemotaxis in vitro. In the present study we investigated the ability of NPS, in comparison with the proinflammatory peptide formyl-Met-Leu-Phe (fMLP), to stimulate human monocyte chemotaxis. At a concentration of 10(-8)M fMLP significantly stimulated chemotaxis. NPS produced a concentration dependent chemotactic action over the concentration range 10(-12) to 10(-5)M. The NPSR antagonists [D-Cys((t)Bu)(5)]NPS, [(t)Bu-D-Gly(5)]NPS and SHA 68 were used to pharmacologically characterize NPS action. Monocyte chemoattractant effect of NPS, but not fMLP, was completely blocked by either peptide antagonists or SHA with the nonpeptide molecule being more potent. None of the NPSR antagonists modified per se random cell migration. Thus, the present study demonstrated that NPS is able to stimulate human monocyte chemotaxis and that this effect is entirely due to selective NPSR activation.
2013
- Skin Wound Healing in Different Aged Xenopus laevis
[Articolo su rivista]
Bertolotti, Evelina; Malagoli, Davide; Franchini, Antonella
abstract
Xenopus froglets can perfectly heal skin
wounds without scarring. To explore whether this
capacity is maintained as development proceeds, we
examined the cellular responses during the repair of skin
injury in 8- and 15-month-old Xenopus laevis. The morphology
and sequence of healing phases (i.e., inflammation,
new tissue formation, and remodeling) were
independent of age, while the timing was delayed in older
frogs. At the beginning of postinjury, wound re-epithelialization
occurred in form of a thin epithelium followed by a
multilayered epidermis containing cells with apoptotic
patterns and keratinocytes stained by anti-inducible nitric
oxide synthase (iNOS) antibody. The inflammatory
response, early activated by recruitment of blood cells immunoreactive
to anti-tumor necrosis factor (TNF)-a,
iNOS, transforming growth factor (TGF)-b1, and matrix
metalloproteinase (MMP)-9, persisted over time. The dermis
repaired by a granulation tissue with extensive angiogenesis,
inflammatory cells, fibroblasts, and anti-a-SMA
positive myofibroblasts. As the healing progressed,
wounded areas displayed vascular regression, decrease in
cellularity, and rearrangement of provisional matrix. The
epidermis restored to a prewound morphology while granulation
tissue was replaced by a fibrous tissue in a scarlike
pattern. The quantitative PCR analysis demonstrated
an up-regulated expression of Xenopus suppressor of cytokine
signaling 3 (XSOCS-3) and Xenopus transforming
growth factor-b2 (XTGF-b2) soon after wounding and
peak levels were detected when granulation tissue was
well developed with a large number of inflammatory cells.
The findings indicate that X. laevis skin wound healing
occurred by a combination of regeneration (in epidermis)
and repair (in dermis) and, in contrast to froglet scarless
wound healing, the growth to a more mature adult stage
is associated with a decrease in regenerative capacity
with scar-like tissue formation.
2013
- The injection of LPS induces epigenetic changes in Pomacea canaliculata neurons. XIII Meeting of the Italian Association of Developmental and Comparative Immunology, Palermo, Inv. Surv. J., 10: 16, 2013
[Abstract in Rivista]
Accorsi, Alice; Rigillo, Giovanna; Malagoli, Davide; Blom, Johanna Maria Catharina; Tascedda, Fabio; Ottaviani, Enzo
abstract
The injection of LPS induces epigenetic changes in Pomacea canaliculata neurons
2013
- Time-related changes of hemocyte morphology in the gastropod Pomacea canaliculata. XIV Meeting of the Italian Association of Developmental and Comparative Immunology, Palermo, Inv. Surv. J., 10: 21, 2013.
[Abstract in Rivista]
Malagoli, Davide; Accorsi, Alice; Bucci, L.; De Eguileor, M.; Ottaviani, Enzo
abstract
Time-related changes of hemocyte morphology in the gastropod Pomacea canaliculata
2012
- Amyloid/Melanin distinctive mark in invertebrate immunity
[Articolo su rivista]
Grimaldi, A.; Girardello, R.; Malagoli, D.; Falabella, P.; Tettamanti, G.; Valvassori, R.; Ottaviani, E.; de Eguileor, M.
abstract
Protostomes and Deuterostomes show the same nexus between melanin production, and amyloid fibril production, i.e., the presence of melanin is indissolubly linked to amyloid scaffold that, in turn, is conditioned by the redox status/cytoplasmic pH modification, pro-protein cleavage presence, adrenocorticotropin hormone (ACTH), melanocyte-stimulating hormone (α-MSH), and neutral endopeptidase (NEP) overexpressions. These events represent the crucial component of immune response in invertebrates, while in vertebrates these series of occurrences could be interpreted as a modest and very restricted innate immune response. On the whole, it emerges that the mechanisms involving amyloid fibrils/pigment synthesis in phylogenetically distant metazoan (viz, cnidaria, molluscs, annelids, insects, ascidians and vertebrates) are evolutionary conserved. Furthermore, our data show the relationship between immune and neuroendocrine systems in amyloid/melanin synthesis. Indeed the process is closely associated to ACTH-α-MSH production, and their role in stress responses leading to pigment production reflects and confirms again their ancient phylogeny.
2012
- Deciphering Insect Immunity: New Insights from RNA Interference
[Monografia/Trattato scientifico]
Malagoli, Davide; Mandrioli, Mauro
abstract
Insect and vertebrate evolutionary histories are separated by more than 500 million years, but the molecular bases of several fundamental biological functions, including innate immune response, seem to have been conserved during metazoan diversification. As a consequence, insects represent good models for gaining new insights into biological basis of human immune-surveillance and pathology. Gene silencing consists in several fruitful techniques, such as the production of loss-of-function mutants and RNA interference. These methods, when applied to models for which molecular databases are available, allow the genetic dissection of several immune-related processes and pathways. In the present review, we will focus on the recent advances on insect immunity derived from the application of gene silencing techniques in Drosophila melanogaster and Anopheles gambiae.
2012
- Drosophila Helical factor is an inducible protein acting as an immune-regulated cytokine in S2 cells.
[Articolo su rivista]
Malagoli, Davide; Accorsi, Alice; Sacchi, Sandro; Basile, Valentina; Mandrioli, Mauro; Pinti, Marcello; D., Conklin; Ottaviani, Enzo
abstract
The innate immunity of Drosophila melanogaster is based on cellular and humoral components. Drosophila Helical factor (Hf), is a molecule previously discovered using an in silico approach and whose expression is controlled by the immune deficiency (Imd) pathway. Here we present evidence demonstrating that Hf is an inducible protein constitutively produced by the S2 hemocyte-derived cell line. Hf expression is stimulated by bacterial extracts that specifically trigger the Imd pathway. In absence of any bacterial challenge, the recombinant form of Hf can influence the expression of the antimicrobial peptides (AMPs) defensin but not drosomycin. These data suggest that in vitro Hf is an inducible and immune-regulated factor, with functions comparable to those of secreted vertebrate cytokines
2012
- Guidelines for the use and interpretation of assays for monitoring autophagy.
[Articolo su rivista]
Klionsky, Dj; Abdalla, Fc; Abeliovich, H; Abraham, Rt; Acevedo Arozena, A; Adeli, K; Agholme, L; Agnello, M; Agostinis, P; Aguirre Ghiso, Ja; Ahn, Hj; Ait Mohamed, O; Ait Si Ali, S; Akematsu, T; Akira, S; Al Younes, Hm; Al Zeer, Ma; Albert, Ml; Albin, Rl; Alegre Abarrategui, J; Aleo, Mf; Alirezaei, M; Almasan, A; Almonte Becerril, M; Amano, A; Amaravadi, R; Amarnath, S; Amer, Ao; Andrieu Abadie, N; Anantharam, V; Ann, Dk; Anoopkumar Dukie, S; Aoki, H; Apostolova, N; Auberger, P; Baba, M; Backues, Sk; Baehrecke, Eh; Bahr, Ba; Bai, Xy; Bailly, Y; Baiocchi, R; Baldini, G; Balduini, W; Ballabio, A; Bamber, Ba; Bampton, Et; Bánhegyi, G; Bartholomew, Cr; Bassham, Dc; Bast RC, Jr; Batoko, H; Bay, Bh; Beau, I; Béchet, Dm; Begley, Tj; Behl, C; Behrends, C; Bekri, S; Bellaire, B; Bendall, Lj; Benetti, L; Berliocchi, L; Bernardi, H; Bernassola, F; Besteiro, S; Bhatia Kissova, I; Bi, X; Biard Piechaczyk, M; Blum, Js; Boise, Lh; Bonaldo, P; Boone, Dl; Bornhauser, Bc; Bortoluci, Kr; Bossis, I; Bost, F; Bourquin, Jp; Boya, P; Boyer Guittaut, M; Bozhkov, Pv; Brady, Nr; Brancolini, C; Brech, A; Brenman, Je; Brennand, A; Bresnick, Eh; Brest, P; Bridges, D; Bristol, Ml; Brookes, Ps; Brown, Ej; Brumell, Jh; Brunetti Pierri, N; Brunk, Ut; Bulman, De; Bultman, Sj; Bultynck, G; Burbulla, Lf; Bursch, W; Butchar, Jp; Buzgariu, W; Bydlowski, Sp; Cadwell, K; Cahová, M; Cai, D; Cai, J; Cai, Q; Calabretta, Bruno; Calvo Garrido, J; Camougrand, N; Campanella, M; Campos Salinas, J; Candi, E; Cao, L; Caplan, Ab; Carding, Sr; Cardoso, Sm; Carew, Js; Carlin, Cr; Carmignac, V; Carneiro, La; Carra, Serena; Caruso, Ra; Casari, G; Casas, C; Castino, R; Cebollero, E; Cecconi, F; Celli, J; Chaachouay, H; Chae, Hj; Chai, Cy; Chan, Dc; Chan, Ey; Chang, Rc; Che, Cm; Chen, Cc; Chen, Gc; Chen, Gq; Chen, M; Chen, Q; Chen, Ss; Chen, W; Chen, X; Chen, X; Chen, X; Chen, Yg; Chen, Y; Chen, Y; Chen, Yj; Chen, Z; Cheng, A; Cheng, Ch; Cheng, Y; Cheong, H; Cheong, Jh; Cherry, S; Chess Williams, R; Cheung, Zh; Chevet, E; Chiang, Hl; Chiarelli, R; Chiba, T; Chin, Ls; Chiou, Sh; Chisari, Fv; Cho, Ch; Cho, Dh; Choi, Am; Choi, D; Choi, Ks; Choi, Me; Chouaib, S; Choubey, D; Choubey, V; Chu, Ct; Chuang, Th; Chueh, Sh; Chun, T; Chwae, Yj; Chye, Ml; Ciarcia, R; Ciriolo, Mr; Clague, Mj; Clark, Rs; Clarke, Pg; Clarke, R; Codogno, P; Coller, Ha; Colombo, Mi; Comincini, S; Condello, M; Condorelli, F; Cookson, Mr; Coombs, Gh; Coppens, I; Corbalan, R; Cossart, P; Costelli, P; Costes, S; Coto Montes, A; Couve, E; Coxon, Fp; Cregg, Jm; Crespo, Jl; Cronjé, Mj; Cuervo, Am; Cullen, Jj; Czaja, Mj; D'Amelio, M; Darfeuille Michaud, A; Davids, Lm; Davies, Fe; De Felici, M; de Groot, Jf; de Haan, Ca; De Martino, L; De Milito, A; De Tata, V; Debnath, J; Degterev, A; Dehay, B; Delbridge, Lm; Demarchi, F; Deng, Yz; Dengjel, J; Dent, P; Denton, D; Deretic, V; Desai, Sd; Devenish, Rj; Di Gioacchino, M; Di Paolo, G; Di Pietro, C; Díaz Araya, G; Díaz Laviada, I; Diaz Meco, Mt; Diaz Nido, J; Dikic, I; Dinesh Kumar, Sp; Ding, Wx; Distelhorst, Cw; Diwan, A; Djavaheri Mergny, M; Dokudovskaya, S; Dong, Z; Dorsey, Fc; Dosenko, V; Dowling, Jj; Doxsey, S; Dreux, M; Drew, Me; Duan, Q; Duchosal, Ma; Duff, K; Dugail, I; Durbeej, M; Duszenko, M; Edelstein, Cl; Edinger, Al; Egea, G; Eichinger, L; Eissa, Nt; Ekmekcioglu, S; El Deiry, Ws; Elazar, Z; Elgendy, M; Ellerby, Lm; Eng, Ke; Engelbrecht, Am; Engelender, S; Erenpreisa, J; Escalante, R; Esclatine, A; Eskelinen, El; Espert, L; Espina, V; Fan, H; Fan, J; Fan, Qw; Fan, Z; Fang, S; Fang, Y; Fanto, M; Fanzani, A; Farkas, T; Farré, Jc; Faure, M; Fechheimer, M; Feng, Cg; Feng, J; Feng, Q; Feng, Y; Fésüs, L; Feuer, R; Figueiredo Pereira, Me; Fimia, Gm; Fingar, Dc; Finkbeiner, S; Finkel, T; Finley, Kd; Fiorito, F; Fisher, Ea; Fisher, Pb; Flajolet, M; Florez McClure, Ml; Florio, S; Fon, Ea; Fornai, F; Fortunato, F; Fotedar, R; Fowler, Dh; Fox, Hs; Franco, R; Frankel, Lb; Fransen, M; Fuentes, Jm; Fueyo, J; Fujii, J; Fujisaki, K; Fuj
abstract
In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
2012
- Methoxyfenozide and pyriproxifen alter the cellular immune reactions of Eurygaster integriceps Puton (Hemiptera: Scutelleridae) against Beauveria bassiana
[Articolo su rivista]
A., Zibaee; A. R., Bandani; Malagoli, Davide
abstract
Effects of the two insect growth regulators (IGRs) methoxyfenozide, 20-hydroxyecdysone (20E) agonist,and pyriproxifen, Juvenile hormone (JH) agonist, were examined on the cellular immune responses of theSunn pest, Eurygaster integriceps versus the entomopathogenic fungus Beauveria bassiana. The simultaneoustreatment with the IGRs and the fungal spores altered haemocyte count (total and differentiate), nodulationresponse and phenoloxidase (PO) activity in a dose- and time-dependent manner. It was observed thatdifferent concentrations of methoxyfenozide increased total and differentiate haemocyte numbers as wellas B. bassiana-induced nodulation response. In contrast with the JH agonist, pyriproxifen significantlydecreased total and differentiate haemocyte numbers and inhibited nodule formation in E. integricepsadults. The 20E agonist displayed major effects when injected at the doses 2.79 and 5.59 lg/mg adult. Incontrast, injecting adults by pyriproxifen significantly impaired their ability to raise an efficacious responseagainst the fungal spores. The ability of the two IGR analogues to interfere with activity of the PO systemin haemolymph of E. integriceps adults was also investigated 6 h after injection by fungal spores.Methoxyfenozide had an excitatory effect on PO activity when the 5.59 lg/mg concentration was usedagainst adults. Conversely, pyriproxifen had an inhibitory effect on PO activity when used at 1.49 lg/mgadult concentration. These findings demonstrate that pyriproxifenmayinterfere with cell-mediated immunityof E. integriceps. So, pyriproxifen could be a good candidate for the integrated control of the Sunn pest.
2012
- TP53 codon 72 polymorphism affects accumulation of mtDNA damage in human cells
[Articolo su rivista]
S., Altilia; A., Santoro; Malagoli, Davide; C., Lanzarini; J. A., Ballesteros Álvarez; G., Galazzo; D. C., Porter; P., Crocco; G., Rose; G., Passarino; I. B., Roninson; C., Franceschi; S., Salvioli
abstract
Human TP53 gene is characterised by a polymorphism at codon 72 leading to an Arginine‐to‐Proline (R/P)substitution. The two resulting p53 isoforms have a different subcellular localisation after stress (more nuclear or moremitochondrial for the P or R isoform, respectively). p53P72 variant is more efficient than p53R72 in inducing the expressionof genes involved in nuclear DNA repair. Since p53 is involved also in mitochondrial DNA (mtDNA) maintenance, wewondered whether these p53 isoforms are associated with different accumulation of mtDNA damage. We observed thatcells bearing p53R72 accumulate lower amount of mtDNA damage upon rotenone stress with respect to cells bearingp53P72, and that p53R72 co‐localises with polymerase gamma more than p53P72. We also analysed the in vivoaccumulation of heteroplasmy in a 300 bp fragment of mtDNA D‐loop of 425 aged subjects. We observed that subjects withheteroplasmy higher than 5% are significantly less than expected in the p53R72/R72 group. On the whole, these datasuggest that the polymorphism of TP53 at codon 72 affects the accumulation of mtDNA mutations, likely through thedifferent ability of the two p53 isoforms to bind to polymerase gamma, and may contribute to in vivo accumulation ofmtDNA mutations.
2012
- The case of the “serfdom” condition of phagocytic immune cells
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; A., Grimaldi; M., de Eguileor
abstract
In a modern immunological perspective, it may be asserted that the phagocytic cell should not be considered as the "serfdom", but rather the pivot of the immune system. Indeed, the invertebrate immunocyte as well as the vertebrate macrophage play a central role in immunity, inflammation and stress response. The evolutionary conserved immune-neuroendocrine effector system have remained of fundamental importance in defense against pathogens, and its efficiency increased through synergy with the new, clonotipical recognition repertoire in vertebrates.
2012
- The immuneregulator role of neprilysin (NEP) in invertebrates
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; Grimaldi, A; Eguileor, M. de
abstract
Neprilysin (NEP) represents an important enzyme in both vertebrates and invertebrates. In the present report we have focused our attention to invertebrates. In particular, a structure related to CD10/NEP as well as its activity in different tissues, such as immunocytes, nervous tissue and muscle of various species were detected. Moreover, the role played by the enzyme in the interactions between host and parasite has also been reported. The findings indicate that NEP immunoregulation is a well-balanced process that, with appropriate physiological and homeostatic responses to challenges, allows the survival and well-being of the species
2012
- The main actors involved in parasitization of Heliothis virescens larva.
[Articolo su rivista]
Grimaldi, A; Tettamanti, G; Congiu, T; Girardello, R; Malagoli, Davide; Falabella, P; Valvassori, R; Ottaviani, Enzo; de Eguileor, M.
abstract
At the moment of parasitization by another insect, the host Heliothis larva is able to defend itself by the activation of humoral and cellular defenses characterized by unusual reactions of hemocytes in response to external stimuli. Here, we have combined light and electron microscopy, staining reactions, and immunocytochemical characterization to analyze the activation and deactivation of one of the most important immune responses involved in invertebrates defense, i.e., melanin production and deposition. The insect host/parasitoid system is a good model to study these events. The activated granulocytes of the host insect are a major repository of amyloid fibrils forming a lattice in the cell. Subsequently, the exocytosed amyloid lattice constitutes the template for melanin deposition in the hemocel. Furthermore, cross-talk between immune and neuroendocrine systems mediated by hormones, cytokines, and neuromodulators with the activation of stress-sensoring circuits to produce and release molecules such as adrenocorticotropin hormone, alpha melanocyte-stimulating hormone, and neutral endopeptidase occurs. Thus, parasitization promotes massive morphological and physiological modifications in the host insect hemocytes and mimics general stress conditions in which phenomena such as amyloid fibril formation, melanin polymerization, pro-inflammatory cytokine production, and activation of the adrenocorticotropin hormone system occur. These events observed in invertebrates are also reported in the literature for vertebrates, suggesting that this network of mechanisms and responses is maintained throughout evolution.
2011
- Cellular immune reactions of the sunn pest, Eurygaster integriceps, to the entomopathogenic fungus, Beauveria bassiana and its secondary metabolites
[Articolo su rivista]
A., Zibaee; A. R., Bandani; R., Talaei Hassanlouei; Malagoli, Davide
abstract
In this study, five morphological types of circulating hemocytes were recognized in thehemolymph of the adult sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae),namely prohemocytes, plasmatocytes, granulocytes, adipohemocytes, and oenocytoids. Theeffects of the secondary metabolites of the entomopathogenic fungus Beauveria bassiana oncellular immune defenses of Eurygaster integriceps were investigated. The results showed thatthe fungal secondary metabolites inhibited phagocytic activity of E. integriceps hemocytes andhampered nodule formation. A reduction of phenoloxidase activity was also observed. The datasuggest that B. bassiana produce secondary metabolites that disable several immune mechanismsallowing the fungus to overcome and then kill its host. This characteristic makes B. bassiana apromising model for biological control of insect pests such as E. integriceps.
2011
- In vitro characterization of the cytokine Drosophila Helical factor
[Abstract in Rivista]
Malagoli, Davide; A., Accorsi; D., Conklin; Filaferro, Monica; Mandrioli, Mauro; Pinti, Marcello; S., Sacchi; Ottaviani, Enzo
abstract
Drosophila Helical factor (Hf) is a protein discovered through the QT method, an algorithm specifically designed for finding helical cytokines. Since in vivo experiments suggested the involvement of Hf in Drosophila melanogaster immunity, we have proceeded with the characterization of Hf functions in the macrophage-like Drosophila embryonic hemocytes, SL2 cell line. qPCR results demonstrated that Hf gene is induced in the SL2 cell line, after either 6 or 24 h incubation with Escherichia coli-purified peptidoglycan. The silencing of Hf expression through RNAi resulted in the reduced capability of synthesizing antimicrobial peptides (AMP) after exposure to heat-inactivated E. coli. The effects of the recombinant peptide rHf have also been tested in the SL2 cell line. rHf promotes the expression and triggers the release of Hf from the hemocytes, and stimulates the synthesis of the antimicrobial peptides (AMP) Defensin and Drosomycin, without any further immune stimulation. Consistent with the output of the QT method, which predicts Hf as a secreted protein, chromatin immune-precipitation experiments confirmed that Hf does not bind DNA, excluding that it acts as an immune-regulated transcription factor. Finally, rHf neither exerts chemotactic action nor triggers bacterial phagocytosis in SL2 cells.Altogether, our data supports the prediction that Hf is an helical cytokine produced and secreted by the hemocytes and it is mainly involved in the regulation of the humoral component of the immune response of D. melanogaster.
2011
- Purification and characterization of phenoloxidase from the hemocytes of Eurygaster integriceps Puton (Hemiptera: Scutelleridae)
[Articolo su rivista]
A., Zibaee; A. R., Bandani; Malagoli, Davide
abstract
Insects protect themselves from microbial invaders by two main immune activities, namely the cellular and humoral reactions. Phenoloxidases are oxidative enzymes that have an important role in both cell-mediated and humoral immunity. In this study, the purification and biochemical characterization of a phenoloxidase from the hemocytes of the Sunn pest Eurygaster integriceps Puton (Hemiptera: Scutelleridae) was carried out. After the final purification step, the enzyme was purified 7.31-fold with a recovery of 3.94% and a specific activity of 4.95 U/mg protein. Results of the biochemical characterization showed that the purified phenoloxidase has a maximum activity at pH 6 and at 30-35 ˚C and is stable for 24-36 hours. Divalent cations such as Ca2+ and Cu2+ significantly increased the enzymatic activity and synthetic inhibitors such as phenylthiourea significantly decreased it. The purified phenoloxidase has a molecular weight of 22 kDa. The current paper represents a further step towards the characterization of humoral immunity of E. integriceps in order to develop new strategies for the biological control of the Sunn pest
2011
- The evolution of proopiomelanocortin: looking for the invertebrate fingerprints.
[Articolo su rivista]
Malagoli, Davide; A., Accorsi; Ottaviani, Enzo
abstract
The presence and role of the pro-opiomelanocortin (POMC) gene and encoded peptides in invertebratesare here summarized and discussed. Some of the POMC-derived peptides show a significant similarityregarding their functions, suggesting their appearance before the split of protostomian–deuterostomianlineages and their maintenance during evolution. The basic mechanisms that govern the exchange ofinformation between cells are usually well conserved, and this could have also been for POMC-derivedpeptides, that are mainly involved in fundamental functions such as immune and neuroendocrineresponses. However, the presence and functions that POMC-derived peptides exhibit in taxonomicallydistant models, are not always reflected by the expected gene homology, leaving the problem of POMCevolution in invertebrates in need of additional study.
2011
- The evolution of the adipose tissue: A neglected enigma
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; Franceschi, Claudio
abstract
The complexity of the anatomical distribution and functions of adipose tissue (AT) has been rarely analyzed
in an evolutionary perspective. From yeast to man lipid droplets are stored mainly in the form of
triglycerides in order to provide energy during periods when energy demands exceed caloric intake. This
simple scenario is in agreement with the recent discovery of a highly conserved family of proteins for fat
storage in both unicellular and multicellular organisms. However, the evolutionary history of organs such
as the fat body in insects, playing a role in immunity and other functions besides energy storage and thermal
insulation, and of differently distributed subtypes of AT in vertebrates is much less clear. These topics
still await a systematic investigation using up-to-date technologies and approaches that would provide
information useful for understanding the role of different AT subtypes in normal/physiological conditions
or in metabolic pathologies of humans.
2010
- Autophagy and its physiological relevance in arthropods: Current knowledge and perspectives.
[Articolo su rivista]
Malagoli, Davide; F. C., Abdalla; Y., Cao; Q., Feng; K., Fujisaki; A., Gregorc; T., Matsuo; I. P., Nezis; I. S., Papassideri; M., Sass; E. C., Silva Zacarin; G., Tettamanti; R., Umemiya Shirafuji
abstract
The autophagic process is one of the best examples of a conserved mechanism of survival in eukaryotes. At the molecular level there are impressive similarities between unicellular and multicellular organisms, but there is increasing evidence that the same process may be used for different ends, i.e., survival or death, at least at the cellular level. Arthropods encompass a wide variety of invertebrates such as insects, crustaceans and spiders, and thus represent the taxon in which most of the investigations on autophagy in nonmammalian models are performed. The present review is focused on the genetic basis and the physiological meaning of the autophagic process in key models of arthropods. The involvement of autophagy in programmed cell death, especially during oogenesis and development, is also discussed.
2010
- Cytokine network in invertebrates: the very next phase of comparative immunology
[Articolo su rivista]
Malagoli, Davide
abstract
Information on invertebrate cytokines has been growing impressively in the past five years. However, molecular characterization of newly discovered cytokines has not proportionally improved our understanding of the main reason underpinning their conservation among metazoans. One possible explanation is that cytokines have been conserved for the fundamental processes they control, but in mammals a single cytokine can hardly be considered as controller of complex reactions. In mammals, cytokines constitute a network of communication, and only this network can be considered the real controller of the effects that cytokines exert on immune or developmental functions. In all, the capability of constituting a network could represent the principal reason for cytokine evolution and conservation through diversification of metazoans
2010
- Discrepant effects of mammalian factors on molluscan cell motility, chemotaxis and phagocytosis: divergent evolution or finely tuned contingency?
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
Cell motility, cell migration and phagocytosis are distinct, though frequently sequential, processes. They are fundamental for the maintenance of homoeostasis in single cells as well as in pluricellular organisms. Like vertebrates, invertebrate immune functions are strictly dependent on cell motility, chemotaxis and phagocytosis. Several comparative immunobiology experiments have tested the effects of mammalian factors on cell migration and phagocytic activity in invertebrate immune-competent cells. The discrepancies that were found suggest various hypotheses, e.g. species-specific reactions to heterologous factors. Here, we reconsider data concerning the effects of POMC (proopiomelanocortin)-derived peptides, cytokines and growth factors on molluscan immunocytes in the light of recent findings that also encompass the effects of experimental conditions.
2010
- Effects of marine toxins on Xenopus laevis early development
[Abstract in Rivista]
Franchini, Antonella; Malagoli, Davide; Ottaviani, Enzo
abstract
Okadaic acid (OA) is a lipophilic compound produced by several marine dinoflagellates, almost exclusively accumulated in mussel digestive gland. The consumption of contaminated animals provokes a syndrome in humans known as diarrheic shellfish poisoning. Palytoxin (PTX) is a large, water soluble polyalcohol found in a variety of marine organisms ranging from dinoflagellates to fish, implicated in seafood poisoning and classified as neurotoxin.
Our experiments were performed by using the Frog Embryo Teratogenesis Assay-Xenopus (FETAX) protocol, and X. laevis embryos at early gastrula stage were treated with different toxin concentrations (0.1, 1 and 10 nM for OA and 0.37, 37 and 370 nM for PTX) for 5 days. The bioassay showed that both toxins affected Xenopus development in terms of mortality, delayed growth and embryo malformation. In particular significant mortality rates were observed with PTX higher dose and the initial sample population decreased by about 80% at assay end. The observation of surviving larvae showed a marked tail folding. Further histological and histochemical studies revealed disorders to the nervous system (the most sensitive tissue) and to the tail skeletal musculature, while alterations also involved the main visceral organs.
Molecular biology-based experiments assessed the expression of four genes (siamois, engrailed-2, bmp4, and myf5) involved in the early events of Xenopus development (stages 11-47) and showed that PTX induces an increase in expression levels in all genes, while the response to OA is stage-dependent, with the embryonic development stages more sensitive than the larval stages. The de-regulated gene expression patterns may account for FETAX and histological data.
2010
- Gene Silencing and the Analysis of Immune Response in Model Insects
[Capitolo/Saggio]
Malagoli, Davide; Mandrioli, Mauro
abstract
Insects are organisms of considerable interest for comparative biology and medicine, therefore it is not surprising that several publications referred to them as model organisms. Insect and vertebrate evolution diverged more than 500 million years ago, but the molecular bases of several fundamental biological functions, including innate immune response, were already established in their common progenitor and have been conserved. Consequently, starting from information collected in insects, new insights into human biology and pathology were gained. Gene silencing includes several powerful methods, such as the production of loss-of-function mutants and RNA interference. These procedures, in particularly when performed in models for which molecular databases are already available, allow the genetic dissection of several immune-related processes and pathways. In the present review, we will concentrate our attention on the information derived from gene silencing techniques on insect immune signalling with particular attention for Drosophila melanogaster and Anopheles gambiae.
2010
- Inflammatory response in molluscs: cross-taxa and evolutionary considerations
[Articolo su rivista]
Ottaviani, Enzo; Franchini, Antonella; Malagoli, Davide
abstract
Inflammation represents the rapid and efficient elimination of damaged tissue and microbes and eventually the restoration oftissue functionality. Inflammatory response is one of the vital reactions to body injury, acting alongside the restoration of homeostasis,wound repair and immune response. In mammals, wound healing is a process that seeks to restore tissue integrity and function, and ischaracterized by a series of biological processes including inflammatory response. Here, we review pioneering experiments and recentobservations in invertebrate models suggesting that in highly divergent and evolutionary distant taxa, such as molluscs, insects and vertebrates,the inflammatory response could be driven by a pool of molecules sharing common evolutionary origin.
2010
- Lectins and cytokines in celomatic invertebrates: Two tales with the same end
[Articolo su rivista]
Malagoli, D.; Sacchi, S.; Ottaviani, E.
abstract
The paper presents the principle data regarding the presence and the roles of lectins and cytokines in invertebrates. The former have been described in the main invertebrate taxa, such molluscs, annelids, arthropods, echinoderms and tunicates, while convincing evidence for cytokines was found only in the insects, Drosophila melanogaster and Pseudaletia separata, and the freshwater crayfish, Pacifastacus leniusculus. Lectins and cytokines share convergent and common functions, and one of the multiples roles of these messenger molecules is their participation in fighting against non-self.
2010
- Life is a huge compromise: Is the complexity of the vertebrate immune-neuroendocrine system an advantage or the price to pay?
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
Recent advances in comparative immunology have established that invertebrates produce hypervariable molecules probably related to immunity, suggesting the possibility of raising a specific immune response. "Priming" and "tailoring" are terms now often associated with the invertebrate innate immunity. Comparative immunologists contributed to eliminate the idea of a static immune system in invertebrates, making necessary to re-consider the evolutive meaning of immunological memory of vertebrates. If the anticipatory immune system represents a maximally efficient immune system, why can it be observed only in vertebrates, especially in consideration that molecular hypervariability exists also in invertebrates? Using well-established theories concerning the evolution of the vertebrate immunity as theoretical basis we analyze from an Eco-immunology-based perspective why a memory-based immune system may have represented an evolutive advantage for jawed vertebrates. We hypothesize that for cold-blooded vertebrates memory represents a complimentary component that flanks the robust and fundamental innate immunity. Conversely, immunological memory has become indispensable and fully exploited in warm-blooded vertebrates, due to their stable inner environment and high metabolic rate, respectively.
2010
- Targets and effects of yessotoxin, okadaic acid and palytoxin: a differential review. Mar. Drugs
[Articolo su rivista]
Franchini, Antonella; Malagoli, Davide; Ottaviani, Enzo
abstract
In this review, we focus on processes, organs and systems targeted by the marine toxins yessotoxin (YTX), okadaic acid (OA) and palytoxin (PTX). The effects of YTX and their basis are analyzed from data collected in the mollusc Mytilus galloprovincialis, the annelid Enchytraeus crypticus, Swiss CD1 mice and invertebrate and vertebrate cell cultures. OA and PTX, two toxins with a better established mode of action, are analyzed with regard to their effects on development. The amphibian Xenopus laevis is used as a model, and the Frog Embryo Teratogenesis Assay-Xenopus (FETAX) as the experimental protocol.
2010
- The common origin of immune and neuroendocrine systems
[Capitolo/Saggio]
Ottaviani, Enzo; Malagoli, Davide
abstract
In this book, the abstract was replaced by the Editor's comments (see Informazioni Utili)
2010
- The state of art on the immune and neuroendocrine functions of corticotrophin-releasing hormone and urocortin in invertebrates
[Capitolo/Saggio]
Ottaviani, Enzo; Malagoli, Davide
abstract
Corticotrophin-releasing hormone (CRH) and urocortin (UCN) are well-known peptides whose functions, in mammals, include different aspects of stress-response, such as the activation of the hypothalamus-hypophysis-adrenal gland axis and the reduction of the appetite, respectively. Expecially in vertebrates, numerous sequences are now available for CRH, UCN and their paralogs that are collectively dubbed as CRH-family members.Together with a better understanding of their functions, the increased availability of CRHfamily member sequences allowed sequence comparison and the consequent flourishing of evolutive hypotheses. Even if problems and conundrums still persist about the origin and the diversification of the CRH-family components, there is growing evidence that this family of peptides, the related receptors and their binding proteins evolved before the split of deuterostomian and protostomian lineages. In invertebrates, CRH-like molecules have been isolated and described mainly in insects whereas no sequence information have so far been derived from other models, such as molluscs or annelids. In insects, the molecules considered as the most probable paralogs of vertebrate CRH-family members are usually ascribed to the group of diuretic hormones. However, in numerous invertebrate taxa, morphological, functional and molecular data indicate the presence of CRH and/or UCN orthologs in circulating and immune-related cell types, that we refer to as immunocytes. Here we review the evidence connecting the presence of invertebrate CRH- and UCN-like peptides to the immune and neuroendocrine system, suggesting that their involvement with the stress response is probably one of their most primitive characteristic, despite the different anatomical scenario within which the stress response occurs in divergent metazoan taxa.
2009
- Around the word stress: its biological and evolutive implications
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide
abstract
Stress is a general adaptive reaction crucial for survival and basically positive that involves the neuroendocrine and the immune systems. In all bilaterian metazoans, the molecular mediators of the stress response, i.e., corticotrophin-releasing hormone, corticotrophin, catecholamines and glucocorticoids, have been preserved during evolution, even if the increased complexity of animals have corresponded to a more articulated stress response that, following the eco-immunology perspective, we speculate to be hierarchically organized along three levels.
2009
- Expression of the genes siamois, engrailed-2, bmp4 and myf5 during Xenopus development in presence of the marine toxins okadaic acid and palytoxin.
[Articolo su rivista]
Franchini, Antonella; Casarini, Livio; Malagoli, Davide; Ottaviani, Enzo
abstract
The present investigation examines the effects of the marine toxins, okadaic acid (OA) and palytoxin (PTX), on some genes involved in the neural and muscular specification and patterning of Xenopus laevis. The RT-PCR analyses performed at different stages of embryonic and larval development (stages 11-47) demonstrated that both toxins induce an over-expression of the genes siamois and engrailed-2 and a different behaviour in bmp4 and myf5. Indeed, OA provoked a significant increase in bmp4 in the earliest stage (11) examined, a down-regulation from stages 12 to 17, and a renewed increase from the beginning of hatching onwards (stages 35-47). In contrast, myf5 was up-regulated in all stages up to 35. PTX induced an over-expression of both bmp4 and myf5 during the embryonic and early larval development stages. The results show that PTX induces an increase in expression levels in all tested genes, while the response to OA seems to be more stage-dependent, with the embryonic development stage more sensitive to the toxin than the larval stages.
2009
- New insights into autophagic cell death in the gypsy moth Lymantria dispar: a proteomic approach
[Articolo su rivista]
Malagoli, Davide; Boraldi, Federica; Annovi, Giulia; Quaglino, Daniela; Ottaviani, Enzo
abstract
Autophagy is an evolutionary ancient process based on the activity of genes conserved from yeast to metazoan taxa. Whereas its role as a mechanism to provide energy during cell starvation is commonly accepted, debate continues about the occurrence of autophagy as a means specifically activated to achieve cell death. The IPLB-LdFB insect cell line, derived from the larval fat body of the lepidoptera Lymantria dispar, represents a suitable model to address this question, as both autophagic and apoptotic cell death can be induced by various stimuli. Using morphological and functional approaches, we have observed that the culture medium conditioned by IPLB-LdFB cells committed to death by the ATPase inhibitor oligomycin A stimulates autophagic cell death in untreated IPLB-LdFB cells. Moreover, proteomic analysis of the conditioned media suggests that, in IPLB-LdFB cells, oligomycin A promotes a shift towards lipid metabolism, increases oxidative stress and specifically directs the cells towards autophagic activity.
2009
- Presence of a low molecular weight lectin in the coelomic fluid of the sea urchin Paracentrotus lividus.
[Articolo su rivista]
Drago, F; Malagoli, Davide; Pezzino, Fm; Durso, V. SAMMARTANO F.
abstract
A low molecular weight (MW) lectin (Paracentrotus lividus small lectin, PlSL) has been found in the sea urchin, Paracentrotus lividus. After gel electrophoresis under denaturing conditions, PlSL exhibits a MW of 13 kDa, while its hemagglutinating activity is Ca2+-independent and inhibited by D-Glucose, L-Rhamnose, D-Arabinose, L-Fucose and N-Acetyl-D-glucosamine. Electrophoretic analysis of the coleomic fluid of P. lividus reveals that the presence of PlSL increases following immune challenge with bacteria, whereas it is annulled as a consequence of osmotic stress. Interestingly, two other putative inducible hemagglutinins of an approximate MW of 11 and 32 kDa were retrieved in concomitance with the stress-promoted disappearance of PlSL.
2009
- Temperature and Ca2+ ion as modulators in cellular immunity of the Sunn pest Eurygaster integriceps Puton (Heteroptera: Scutelleridae)
[Articolo su rivista]
A., Zibaee; A. R., Bandani; R., Talaei Hassanlouei; Malagoli, Davide
abstract
Environmental conditions in addition to divalent cations may affect the interactionsbetween pathogens and insects. Elucidation of factors which modulate insectimmune responses could be a significant part of investigations in this area. In thisstudy, adults of Eurygaster integriceps, as the destructive pest of wheat, were keptat different temperatures in addition to injection with different concentrations ofCa2+ to find the effect on cellular immune reactions against Beauveria bassiana.Results showed that total and differentiate hemocyte numbers, nodule formationand phenoloxidase activity increased with elevation of temperature so that thehigher values were obtained at 30 and 40°C at various intervals. Higher concentrationsof Ca2+ ion (5 mM) caused an increase in plasmatocyte length and widthespecially after 60 min. Similar results were observed for nodule formation andphenoloxidase activity of E. integriceps adults after injection by B. bassiana sporesand phenoloxidase activity. It is clear from the current study that thermoregulationand Ca2+ ion can positively affect the hemocyte numbers especially plasmatocytesand granulocytes, nodule formation and phenoloxidase activity in E. integriceps.The understanding of modulators of the insect immune response may directlyinfluence novel approaches to obtain safe and effective biological control agents.
2009
- The invertebrate blueprint of the connection between aging and immune neuroendocrine responses
[Articolo su rivista]
Ottaviani, E.; Malagoli, D.; Franceschi, C.
abstract
The present paper summarizes the main findings related to aging and immune neuroendocrine responses in invertebrates. In particular, the functional aspects and the genes involved are examined. A possible mechanism of correlation between aging and functioning of immune-neuroendocrine-related genes is also discussed.
2008
- Cell death in the IPLB-LdFB insect cell line: Facts and implications
[Articolo su rivista]
Malagoli, Davide
abstract
Abstract: The present review summarizes findings on stress-induced cell death in the IPLB-LdFB insect cell line derived from the larvalfat body of the lepidopteron Lymantria dispar. Apoptotic, oncotic and autophagic cell death have been described in these cells as a consequenceof oxidative stress or ATP deprivation, and similarities between IPLB-LdFB and mammalian apoptotic pathways have beenhighlighted. Furthermore, starting from observations in the IPLB-LdFB cells, a link has been surmised between relevance of autophagiccell death and developmental processes in the metazoan taxa.
2008
- Cytotoxic activity by the mussel Mytilus galloprovincialis and the Venus clam Chamela gallina in the Adriatic sea in 2007
[Articolo su rivista]
Malagoli, Davide; Casarini, Livio; F., Fiori; Ottaviani, Enzo
abstract
Given the ecological and economic importance of bivalve molluscs, the evaluation of their welfare is one of the primary aims for both biologists and people working in shell fishing. After a three year-long period monitoring the cytotoxic activity exerted by the hemolymph from the mussel Mytilus galloprovincialis, we have concluded that cytotoxicity represents a useful parameter to evaluate the status of the immune activity and therefore the health of mussels in a specific period of the year. During 2007, we compared the mussel cytoxicity with that of the Venus clam Chamelea gallina from contiguous areas of the Northern Adriatic Sea. Our observations indicate that the cytotoxicity of the hemolymph of the two species follows a similar course during the year, suggesting that cytotoxic activity is primarily determined by the life/reproductive cycles.
2008
- Ecoimmunology: is there any room for the neuroendocrine system?
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; M., Capri; C., Franceschi
abstract
Ecological Immunology assumes that immunologicaldefenses must be minimized in terms of cost (energyexpenditure). To reach this goal, a complex and stilllargely unexplored strategy has evolved to assuresurvival. From invertebrates to vertebrates, an integratedimmune–neuroendocrine response appears to be crucialfor the hierarchical redistribution of resources within thebody according to the specific ecological demands.Thus, on the basis of experimental data on the intimaterelationship between stress and immune responses thathas been maintained during evolution, we argue that abroader perspective based on the integration of immuneand neuroendocrine responses should be adopted todescribe the comprehensive strategy that the bodyutilizes to adapt to dynamic environmental conditions.We discuss the hypothesis that a bow-tie architecturemight be suitable to describe the variety of immune–neuroendocrine inputs that continuously target cells andorgans while, at the same time, fulfilling the basicrequirement of minimizing the cost of immune–neuroendocrineresponses. Bow-tie architectures are able toconvert a variety of stimuli (fan in) into a wide range offine-tuned responses (fan out) by passing through theintegrating activity of a core (knot) constituted by alimited number of elements. Finally, we argue thatthe ecologically negotiated immune–neuroendocrinestrategies may have deleterious effects in the postreproductiveperiod of life when, at least in humans, chronic, low-grade, systemic inflammation develops, in accordance with the antagonistic pleiotropy theory of aging.
2008
- Effects of the marine toxins okadaic acid and palytoxin on mussel phagocytosis.
[Articolo su rivista]
Malagoli, Davide; Casarini, Livio; Ottaviani, Enzo
abstract
The present study analyzes the effects of the marine toxins okadaic acid (OA) and palytoxin (PTX) on the phagocytic activity of immunocytes from the mussel Mytilus galloprovincialis.In particular, we describe how the effects of the two biotoxins are influenced by the temperature and experimental stress applied before hemolymph withdrawal. The collected data indicate that OA increases phagocytic activity only when hemolymph incubation is performed at 25 C, but not at 20 C, suggesting a certain degree of dependence of OA effects from the status of mussel immunocytes. Conversely, PTX plays an active role in immunocyte signalling transduction pathways, increases the phagocytic activity and markedly promotes the involvement of p38 mitogen-activated protein (MAP) kinase in phagocytosis. Overall, we conclude that both OA and PTX influence mussel phagocytic activity, and the toxic effects may depend on both the mollusc conditions and the activation of specific signalling pathways.
2008
- Erratum to: Oligomycin A and the IPLB-LdFB insect cell line: Actin and mitochondrial responses [32 (2) 287-292] (DOI:10.1016/j.cellbi.2007.10.011)
[Articolo su rivista]
Tettamanti, G.; Malagoli, D.; Ottaviani, E.; de Eguileor, M.
abstract
2008
- Immune-neuroendocrine biology of invertebrates: A collection of methods
[Articolo su rivista]
Ballarin, L.; Cammarata, M.; Cima, F.; Grimaldi, A.; Lorenzon, S.; Malagoli, D.; Ottaviani, E.
abstract
In the last decade there has been a considerable increase of interest towards the elucidation of several aspects of invertebrate biology, including immunity and neuroendocrinology. However, due to the difficulties connected to the great variety of morphology and adaptations displayed by invertebrates, and also in consideration of the number of techniques that are applied in the various laboratories, research on invertebrates still suffers from hampering that have been substantially overcome in vertebrate models, especially in mammals. The aim of this Technical Report is to provide the reader a useful list of well-established morphological and morpho-functional protocols in order to facilitate the design and make more homogeneous the realization of experiments in the field of invertebrate immune-neuroendocrinology.
2008
- In vitro methods to monitor autophagy in Lepidoptera.
[Capitolo/Saggio]
Tettamanti, G; Malagoli, Davide
abstract
Autophagy is attracting growing interest, especially in relation to increasing evidence of the importance of autophagic processes in animal development, as well as in human cancer progression. In holometabolous insects (i.e., that undergo four distinct life cycle stages, including embryo, larva, pupa and imago), such as flies, butterflies, bees and beetles, autophagy has been found to play a fundamental role in metamorphosis, and given the high degree of conservation of the genes and the basic mechanisms of autophagy, attention to these relatively simple models has increased significantly. Together with Drosophila, Lepidoptera larvae are among the most common invertebrate models in studies concerning the protective action of starvation-induced autophagy or the possible role of autophagy as a programmed cell death process. In this chapter, we provide experimental methods developed for, or applicable to, the study of the autophagic process in the IPLB-LdFB cell line derived from the fat body of the caterpillar of the gypsy moth, Lymantria dispar.
2008
- Oligomycin A and the IPLB-LdFB insect cell line: Actin and mitochondrial responses
[Articolo su rivista]
G., Tettamanti; Malagoli, Davide; Ottaviani, Enzo; M., De Eguileor
abstract
Oligomycin A, an inhibitor of mitochondrial ATP synthase, provokes simultaneous and different responses in IPLB-LdFB insect cell line. The oligomycin A treatment causes mitochondrial loss, increase in reactive oxygen species (ROS), destabilization/reorganization of the actin microfilaments and, finally, autophagic cell death. We speculate that oligomycin A affects the mitochondria and that the impairment of theseorganelles leads to the generation of ROS in quantities that exceed the antioxidant capacity of the cell. This in turn would lead to a feedback loop of increased mitochondrial impairment, amplification of ROS production and the removal of damaged organelles through autophagy.
2008
- hrIL-8 stimulates unpaired (upd)-3 expression and other immune-related activities in Drosophila melanogaster SL2 cell line
[Relazione in Atti di Convegno]
Malagoli, Davide; Sacchi, S; Ottaviani, Enzo
abstract
cellular and humoral components. Among humoral factors, there is less information on soluble molecules able to act as signals during the immune response (i.e. cytokines). To date, only a few cytokines have been observed in different arthropod taxa such as insects (Spätzle and Unpaired [Upd]-3) and crustaceans (astakine). As it has been proposed that a possible involvement of chemotactic factors may increase the expression of upd-3, we have studied the effects of human recombinant ilnterleukin-8 (hrIL-8) on the immune functions of Drosophila melanogaster SL2 macrophage-like cells. We have found that hrIL-8 enhances the expression of upd-3 and of the putative cytokine, Drosophila helical factor (dhf). Furthermore, hrIL-8 promotes the transcription of the antimicrobial peptide genes defensin and cecropin A1. Beside these evidences on humoral factors, hrIL-8 also increases the phagocytic activity of SL2 cells. Our data suggest the existence in D. melanogaster of one or more soluble factors that possibly share some structural similarity with IL-8, eliciting an immune response involving simultaneously cellular and humoral components.
2008
- unpaired (upd)-3 expression and other immune-related functions are stimulated by interleukin-8 in Drosophila melanogaster SL2 cell line
[Articolo su rivista]
Malagoli, Davide; Sacchi, S; Ottaviani, Enzo
abstract
Invertebrate innate immunity relies on both cellular and humoral components. Among humoral factors,there is less information on soluble molecules able to act as signals during the immune response (i.e.cytokines). In Drosophila melanogaster, the cytokine Unpaired (Upd)-3, is known to activate the JAK/STATpathway, but it is still not clear which molecules and pathways are responsible for its induction andsecretion. It has been proposed that highly chemotactic factors may increase the expression of upd-3.In this respect, we have studied the effects of the chemotactic human recombinant (hr) interleukin(IL)-8 on the immune functions of Drosophila SL2 macrophage-like cells. The hrIL-8 increases the percentageof phagocytic cells with a specific timing and enhances the expression of the cytokine, upd-3, as wellas that of the putative cytokine Drosophila helical factor (dhf). The antimicrobial peptides defensin, cecropinA1 and diptericin, are all influenced in their expression by the human chemokine, while hrIL-8 leavesunaffected the expression of drosomycin, i.e. the antimicrobial peptide more strictly connected with theToll pathway. Similar effects to those registered for hrIL-8 are also provoked by a specific activator of theImd pathway, i.e. the Escherichia coli peptidoglycan. RNAi experiments demonstrated that the silencing ofthe Imd pathway-associated kinase dTAK1, leaves unaffected the induction of upd-3, while it completelyabolishes the effects of hrIL-8-on the expression of dhf. Our data suggest that the Imd pathway is not fundamentalin regulating the levels of upd-3, whereas it controls the expression of the putative cytokine dhf.
2007
- A full-length protocol to test hemolytic activity of palytoxin on human erythrocytes
[Articolo su rivista]
Malagoli, Davide
abstract
The hemolytic assay protocols currently utilized to test the presence of the marine biotoxin palytoxin (PTX) are deeply analyzed. In some points, slight modifications and rearrangements have been realized, to obtain an exhaustive protocol suitable to test PTX activity on human erythrocytes.
2007
- A putative helical cytokine functioning in innate immune signalling in Drosophila melanogaster
[Articolo su rivista]
Malagoli, Davide; D., Conklin; Sacchi, Sandro; Mandrioli, Mauro; Ottaviani, Enzo
abstract
In invertebrates and vertebrates, innate immunity is considered the first line of defense mechanism against non-self material. In vertebrates,cytokines play a critical role in innate immune signalling. To date, however, the existence of genes encoding for invertebrate helical cytokines hasbeen anticipated, but never demonstrated. Here, we report the first structural and functional evidence of a gene encoding for a putative helicalcytokine in Drosophila melanogaster. Functional experiments demonstrate that its expression, as well as that of the antimicrobial factors defensinand cecropin A1, is significantly increased after immune stimulation. These observations suggest the involvement of helical cytokines in the innateimmune response of invertebrates.
2007
- Common evolutionary origin of the immune and neuroendocrine systems: from morphological and functional evidences to in silico approaches
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; C., Franceschi
abstract
Ten years ago, we surmised that there was a commonevolutionary origin for the immune and neuroendocrinesystems. This was based on morphological and functionalresults indicating that a common pool of moleculesis shared by the two systems, both ininvertebrates and vertebrates. In the past decade,numerous molecular biology experiments have confirmedsequence similarity between invertebrate andvertebrate neuroimmune mediators, such as corticotrophin-releasing hormone. However, sequence similaritybasedapproaches were inadequate for analyzing otherimmune-related molecules, such as helical cytokines.This review covers older, and more recent findings oninvertebrate immune- and neuroendocrine-related moleculeswith an evolutionary perspective, and suggeststhat protein-folding recognition algorithms are a fundamentaltool in understanding the evolution of immuneandneuroendocrine-related molecules.
2007
- Detection of HSP27-like molecules in the annelid Enchytraeus japonensis after exposure to extremely low frequency magnetic fields (50 Hz).
[Articolo su rivista]
Malagoli, Davide; Marchetti, M; Gobba, Fabriziomaria
abstract
The immunocytochemical study performed on the annelid Enchytraeus japonensis revealed the presence of immunoreactive heat shock protein (HSP)27 molecules in different areas of the body. Positivity was observed in coelomocytes and in epithelial cells of the intestine wall. The exposition of the animals to 400 μT magnetic fields (50 Hz) for 30 min provoked an increased immunoreactivity in some specimens, but after immunoblot experiments, no significant differences in the total content of HSP27-like molecules were found between exposed and non-exposed animals.
2007
- Evaluation of the effects of the marine toxin okadaic acid by using FETAX assay
[Articolo su rivista]
Casarini, Livio; Franchini, Antonella; Malagoli, Davide; Ottaviani, Enzo
abstract
The Frog Embryo Teratogenesis Assay Xenopus (FETAX), is a screening assay using embryos at gastrula stage of the anuran Xenopus laevis to identify substances that may pose a developmental hazard in humans. The FETAX assay evaluates three parameters, i.e. mortality, delayed growth and embryo malformation. In the present investigation, the FETAX protocol was applied to the marine toxin okadaic acid (OA) and the experiments show that OA affects the above parameters in a dose-correlated manner. The morpho-functional modifications induced in embryo organs by OA were also studied. The nervous system, tail skeletal musculature, intestine and kidney appeared particularly damaged, with the former being the most sensitive. On the whole, various advantages emerge from using the FETAX assay: different parameters can be tested simultaneously, the indication of the presence of a potentially dangerous substance is rapid and the assay is a valid alternative to mammalian systems. (c) 2007 Elsevier Ireland Ltd. All rights reserved.
2007
- Evolution game: which came first, the receptor or the ligand?
[Articolo su rivista]
Mandrioli, Mauro; Malagoli, Davide; Ottaviani, Enzo
abstract
On the basis of a bioinformatic approach, we suggest that in invertebrates many ligands interact with a single, ancestral and generalized receptor driving ligand evolution. In vertebrates, on the other hand, the occurrence of gene/genome duplications induced the shift to a ligand-directed evolution of receptors.
2007
- Evolution of helical cytokines: a structural approach
[Relazione in Atti di Convegno]
Malagoli, Davide; Ottaviani, Enzo
abstract
Cytokines are small soluble factors retrieved in mammals and involved in several processes such as immunity and development. They are typically characterized by pleiotropicity, functional and receptor redundancy. In consideration of functional parallels between mammalian and invertebrate immunity, a lot of experiments have been dedicated to the unravelling of cytokine network in both protostomian and deuterostomian invertebrates. The presence of cytokine-like molecules has been evidenced by several morphological and functional investigations in different taxa of invertebrates, leading to the hypothesis that cytokines are molecules of ancient origin, present in metazoans before the division of protostomian and deuterostomian phyletic lines. However, all the recent molecular biology advances indicate that no sequence similarity can be retrieved between the known vertebrate cytokines and the whole genome of invertebrate species. On these basis, functional convergence has been proposed between
25
vertebrate and invertebrate cytokines. The functional convergence would be due to the lectin-like activity of vertebrate cytokines that can be retrieved also in some invertebrate molecules.
In order to unravel this unsolved matter, we have adopted a new bioinformatics approach able to isolate proteins whose structure is comparable to that of mammalian helical cytokines from EST and protein databases. Through this method we have isolated a molecule from Drosophila melanogaster databases that presents the structural characteristics of a helical cytokine (Drosophila helical factor, DHF). Functional experiments performed on third instars larvae and SL2 embryonic hemocyte cell line of D. melanogaster demonstrated that DHF expression was increased after different immune challenges.
From the present findings, it emerges that the contradiction between the amount of morphological and functional evidences and the absence of any homology between mammalian and invertebrate cytokines, could be explained by evolution of cytokine genes thereby conserving specific protein structures rather than amino acid or nucleotide sequences.
From the present findings, it emerges that the contradiction between the amount of morphological and functional evidences and the absence of any homology between mammalian and invertebrate cytokines, could be explained by evolution of cytokine genes thereby conserving specific protein structures rather than amino acid or nucleotide sequences.
2007
- Helical cytokines and invertebrate immunity: a new field of research
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
Description of the invertebrate elical cytokine
2007
- Monitoring of the immune efficiency of Mytilus galloprovincialis in Adriatic sea mussel farms in 2006: regular changes of cytotoxicity during the year
[Articolo su rivista]
Malagoli, Davide; Casarini, Livio; Ottaviani, Enzo
abstract
By monitoring the course of hemolymph cytolytic activity in Mytilus galloprovincialis during 2006, we have observed important fluctuations in the percentage of cytotoxic animals over the year. The changes seem to be correlated with seasonal variations in the temperature, but observations in mussels kept in aquaria indicated that this parameter is not the main cause of the fluctuations. Data presented here suggest that normal levels of cytotoxicity can be predicted in a population for a specific period of the year, therefore confirming the value of this parameter in determining the immune efficiency of mussels at a given time.
2007
- Presence of and stress-related changes in urocortin-like molecules in neurons and immune cells from the mussel Mytilus galloprovincialis
[Articolo su rivista]
Malagoli, Davide; I., Di Paolo; Ottaviani, Enzo
abstract
The distribution of urocortin (UCN)-like material is investigated in the bivalve molluscMytilus galloprovincialis. Immunocytochemical data demonstrate that UCN-like moleculesare present in ganglionic neurons, microglial cells and immunocytes. Moreover, a colocalizationof UCN- and corticotrophin-releasing hormone (CRH)-like molecules is foundin microglial cells and in immunocytes, but not in neurons. Following high salinity-stress experiments, immunoreactivity for UCN and CRH increased in ganglionic neurons and immunocytes. Our findings extend the number of molecules potentially used by molluscan immunocytes to confront stress situations and strengthen the idea of functional conservation of stress-related molecules during evolution.
2007
- Stress and immune response in the mussel Mytilus galloprovincialis
[Articolo su rivista]
Malagoli, Davide; Casarini, Livio; Sacchi, Sandro; Ottaviani, Enzo
abstract
The present study investigates the effects on immune-related parameters of various stress factors (air exposure, mechanicalstress, high temperature and extreme salinity conditions) faced by the bivalve mollusc Mytilus galloprovincialis during marketingprocedures. We observed that some stress typologies increase phagocytosis and the number of circulating immunocytes, whileothers can modify immunocyte response towards a further perturbation, i.e. the marine algal toxin yessotoxin. Our results suggest that non-lethal stress can be counteracted for sometime by increasing the level of some defence parameters. Moreover, our data indicate that fishing and transport procedures could interfere with mussel immunosurveillance.
2006
- 50 Hz magnetic fields of constant or fluctuating intensità: effects on immunocyte HSP 70 in the mussel Mytilus galloprovincialis
[Articolo su rivista]
Malagoli, Davide; Gobba, Fabriziomaria; Ottaviani, Enzo
abstract
Effects of a single 30 min exposure to a 50 Hz, 400 mu T sinusoidal extremely low frequency magnetic field (ELF MF) on hsp70 expression in immunocytes from the bivalve Mytilus galloprovincialis was investigated using RT-PCR and immunoblot approaches. The results indicate that in M. galloprovincialis immunocytes hsp70 expression was unaltered at both transcriptional and translational level after exposure to constant or fluctuating intensity MFs. Bioelectromagnetics 27:427-429, 2006.
2006
- Algal toxin yessotoxin signalling pathways involve immunocyte mussel calcium channels
[Articolo su rivista]
Malagoli, Davide; Casarini, Livio; Ottaviani, Enzo
abstract
A fragment of a putative L-type Ca2+ channel has been identified by molecular biology experiments in immunocytes from the mussel Mytilus galloprovincialis. Using the cell permeable and Ca2+-specific fluorochrome FURA 2-AM, we have demonstrated that the algal toxin yessotoxin (YTX) is able to increase intracellular Ca2+ concentration in M. galloprovincialis immunocytes. The YTX effect on Ca2+ increase is inhibited by the L-type Ca2+ channel inhibitor, verapamil, which is cAMP- and cGMP-dependent; but PKA- and nitric oxide-independent. On the basis of these observations, a possible role for YTX as a potential disturber of mussel immune efficiency is suggested. (c) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.
2006
- Cytokines and invertebrates: TGF-β and PDGF
[Articolo su rivista]
Franchini, Antonella; Malagoli, Davide; Ottaviani, Enzo
abstract
The review outlines the presence and function of TGF-beta and PDGF family members in invertebrates. TGF-beta and PDGF play an important role in development, in immune and neuroendocrine responses and in the wound repair by activating the classical transduction pathways. Generally speaking, these cytokines appear very early in evolution and conserve their functions.
2006
- Growth factors and chemokines: A comparative functional approach between invertebrates and vertebrates
[Articolo su rivista]
Tettamanti, G; Malagoli, Davide; Benelli, R; Albini, A; Grimaldi, A; Perletti, G; Noonan, Dm; de Eguileor, M; Ottaviani, Enzo
abstract
Growth factors and cytokines control and coordinate a broad spectrum of fundamental cellular functions, and are evolutionarily conserved both in vertebrates and invertebrates. In this review, we focus our attention on the functional phylogenetic aspects of growth factors/cytokines like the Transforming Growth Factor-beta (TGF-beta), the Connective Tissue Growth Factor (CTGF), and the Vascular Endothelial Growth Factor (VEGP). We will also delve into the activites of two chemokine families, interleukin (IL)-8 (or CXCL8) and CC chemokine ligand 2/monocyte chemoattractant protein-1 (CCL2). These molecules have been selected for their involvement in immune responses and wound healing processes, where they mediate and finely regulate various regeneration processes like angiogenesis or fibroplasia, not only in vertebrates, but also in invertebrates.
2006
- Lysosomes as the target of yessotoxin in invertebrate and vertebrate cell lines
[Articolo su rivista]
Malagoli, Davide; Marchesini, Erika; Ottaviani, Enzo
abstract
The toxic effects of the algal polyether phycotoxin yessotoxin (YTX) are studied in the insect fat body IPLB-LdFB and the mouse fibroblast NIH3T3 cell lines. Our experiments confirm the cytotoxic action exerted by the toxin in both insect and mammalian cells, but morphological observations, TUNEL experiments and electrophoretic evalution of DNA integrity failed to evidence a clear pro-apoptotic role for YTX. In both IPLB-LdFB and NIH3T3 cell lines, neutral red and acridine orange stainings, together with evaluation of acid phosphatase activity demonstrate that YTX first damages lysosomal vesicles. This is then followed by a progressive depolymerization of actin microfilaments, as shown by phalloidin fluorescent immunostaining. Overall, our data identify in early lysosomal damage and the subsequent cytoskeletal disruption two common steps related to YTX toxicity towards metazoan cells. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
2006
- Monitoring of the immune efficiency of Mytilus galloprovincialis in Adriatic sea mussel farms in 2005
[Articolo su rivista]
Malagoli, Davide; Casarini, Livio; Ottaviani, Enzo
abstract
The monthly evaluation of the cytotoxicity of hemolymph from the mussel Mytilus galloprovincialis
revealed some variations in the percentage of cytotoxic animals during the year. Cytotoxicity is confirmed
to be a dynamic parameter that can be used as an indicator of immune efficiency and, therefore, of the
state of health of the animals.
2006
- Oligomycin A induces autophagy in the IPLB-LdFB insect cell line
[Articolo su rivista]
G., Tettamanti; Malagoli, Davide; Marchesini, Erika; T., Congiu; M., de Eguileor; Ottaviani, Enzo
abstract
Functional and morphological modifications in the IPLB-LdFB insect cell line were examined following a short treatment with a reversible inhibitor of mitochondrial ATP synthase, oligomycin A, and subsequent incubation for various times in oligomycin-A-free medium. Oncosis, apoptosis and autophagy at variable percentages were observed under the various experimental conditions. Together with oncotic and apoptotic pathways that lead directly to cell death, the insect cells responded to ATP depletion with autophagy. Our results revealed that, in most cases, autophagy failed to restore cellular homeostasis, probably because of a massive sequestration of mitochondria in autophagic vacuoles. This critical event was a point of no return and ultimately resulted in cell necrosis. However, cells with a misshapen body and nucleus resembling resistant forms were observed at the end of the experiments. Our findings indicate that oligomycin-A-induced autophagy can promote cell protection or cell destruction and is an open-ended process that can lead to survival or death depending on a combination of concomitant factors.
2005
- Cell-death mechanisms in the IPLB-LdFB insect cell line: a nuclear located Bcl-2-like molecule as a possible controller of 2-deoxy-D-ribose-mediated DNA fragmentation
[Articolo su rivista]
Malagoli, Davide; I., Iacconi; Marchesini, Erika; Ottaviani, Enzo
abstract
In the IPLB-LdFB insect cell line, oncosis and apoptosis are the two pre-mortal processes, whereas necrosis is the post-mortem condition. As found in mammals, adenosine triphosphate depletion of insect cells by oligomycin A induces oncosis. The apoptotic inducer 2-deoxy-D-ribose (dRib) provokes cell death through an intrinsic apoptotic pathway similar to that observed in mammalian models and results in oligonucleosomal DNA fragmentation. The addition to insect cells of an anti-Bcl-2 polyclonal antibody known to prevent dRib-mediated apoptosis abolishes DNA fragmentation, whereas cytochrome c release and the increase in a caspase 3-like activity are still detectable. These and previous findings suggest a double role for the Bcl-2-like molecule in IPLB-LdFB, i.e. the maintenance of mitochondrial integrity and the control of apoptotic machinery at the nuclear level.
2005
- Cytotoxicity as a marker of mussel health status
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
Cytotoxic activity of the mollusc Mytilus galloprovincalis haemolymph was considered as a possible marker of mussel health status. An easy-to-use and low-cost cytotoxicity test was set up to assess whether cytotoxicity is influenced by environmental stress such as an increase in water temperature or a reduction in water pH or salinity. Experiments demonstrated that, despite great variability in cytotoxic activity among the mussel populations, changes in the water physical conditions reduce mussel cytotoxicity, making animals more vulnerable to pathogen aggression.
2005
- Investigation of the loss of byssus in Mytilus galloprovincialis from mussel farms in the Adriatic Sea
[Articolo su rivista]
Franchini, Antonella; Malagoli, Davide; Ottaviani, Enzo
abstract
A fungal infection has been found in the mussel Mytilus galloprovincialis from Adriatic Sea mussel farms. The infection ultimately results in the loss of the byssus, with serious consequences for mussel farming yield. The pathogen provokes the progressive destruction of the foot muscles, also damaging related structures such as the intra-organism part of the byssus apparatus, resulting in loss of the thread component. The affected health status of the animal is also sustained by modifications in the digestive gland structure, ranging from hyperactivity to extreme cell death in the tubula. At present, the identity of the harmful fungus is unknown.
2004
- 50 Hz magnetic fields activate mussel immunocyte p38 MAP kinase and induce HSP70 and 90
[Articolo su rivista]
Malagoli, Davide; M., Lusvardi; Gobba, Fabriziomaria; Ottaviani, Enzo
abstract
Fifty hertz magnetic fields (MFs) induced the expression of heat shock proteins (HSPs) 70 and 90 in immunocytes of the mussel Mytilus galloprovincialis. Animals exposed at 300 muT for three different times (30; 2 X 30; 3 X 30 min), did not show differences in the HSP densitometric values in comparison with non-exposed mussels. At 400 muT, exposed animals showed a time-dependent increase in HSP expression as revealed by Western blot. After exposure to 600 muT, the HSP densitometric values were significantly higher than controls but not related to exposure duration. The induction of HSPs is concomitant with the activation of p38 MAP kinase signalling pathway. The present findings suggest the possibility to modulate the expression of HSPs by an appropriate time-intensity magnetic field exposure.
2004
- Invertebrate humoral factors: cytokines as mediators of cell survival
[Capitolo/Saggio]
Ottaviani, Enzo; Malagoli, Davide; Franchini, Antonella
abstract
The presence and the different functional aspects of cytokine-related molecules in invertebrates are described. Cytokine-like factors affect immune functions, such as cell motility, chemotaxis, phagocytosis and cytotoxicity. In particular, cell migration shows a species-specific effect for IL-1alpha and TNF-alpha and a dose-correlated effect for IL-8, PDGF-AB and TGF-beta1. Apart from some exceptions, the phagocytic effect increases significantly at all the concentrations tested and with all the species used. PDGF-AB, TGF-beta1 and IL-8 provoke conformational changes in mollusk immunocytes, involving the signaling transduction pathways of phosphatidylinositol and cAMP. PDGF-AB and TGF-beta1 partially inhibit the induced programmed cell death in an insect cell line, and the survival effect is mediated by the activation of phosphatidylinositol 3-kinase, PKA and PKC. The exogenous administration of these growth factors in an invertebrate wound repair model showed that they are able to control the wound environment and promote the repair process by accelerating the coordinated activities involved. Moreover, IL-1alpha, IL-2 and TNF-alpha are able to induce nitric oxide synthase. PDGF-AB and TGF-beta1 provoke an increase in neutral endopeptidase-24.11 (NEP)-like activity in membrane preparations from mollusk immunocytes, while NEP deactivates the PDGF-AB- and TGF-beta1-induced cell shape changes. Cytokines are also involved in invertebrate stress response in a manner extremely similar to that in vertebrates. Several studies suggest the existence on the mollusk immunocyte membrane of an ancestral receptor capable of binding both IL-2 and CRH. Furthermore, the competition found between CRH and a large number of cytokines supports the idea that invertebrate cytokine receptors show a certain degree of promiscuity. The multiple functions of cytokines detected in invertebrates underline another characteristic of mammalian cytokines, i.e. their great pleiotropicity. Altogether, the studies on the function of the invertebrate humoral factors show a close overlapping with those found in vertebrates, and the hypothesized missing correlation between invertebrate and vertebrate cytokine genes that is emerging from the limited molecular biology data present in literature might represent a very peculiar strategy followed by Nature in the evolution of cytokines.
2004
- ProCRH in the teleost Ameiurus nebulosus: gene cloning and role in LPS-induced stress response
[Articolo su rivista]
Malagoli, Davide; Mandrioli, Mauro; Ottaviani, Enzo
abstract
The procorticotrophin-releasing hormone (proCRH) gene from the teleost Ameiurus nebulosus was cloned by direct and inverse PCR-based technologies and characterized. The structure of the proCRH gene shows the presence of four exons and three introns giving a total length of 1416 bp. Sequence similarity with the corresponding proCRH coding sequences in Tilapia mossambica and Homo sapiens is 97.7 and 78%, respectively. Western blot experiments performed with an anti-human CRH (1-41) antibody revealed the presence of an immumoreactive molecule with an approximate MW of 18 kDa, a value comparable to that of the putative catfish proCRH peptide. These data suggest that proCRH could be active in A. nebulosus without any cleavage. ProCRH immunoreactive molecules were found in the central nervous system (CNS) and were immunocytochemically detected in the head kidney and in the pancreatic gland. Western blot and immunocytochemical experiments showed an increase in proCRH expression in the CNS after 15 min but not after 120 min exposure to LPS. In contrast, the increased immunopositivity was detectable in the pancreas only after 120 min of treatment, but in the head kidney throughout the entire period of exposure. Our findings indicate that the CNS responds to the altered conditions for a shorter period of time than the peripheral organs, suggesting a hierarchical and time-regulated stress response. However, an independent response in the peripheral organs cannot be excluded in this scenario.
2004
- The effects of parasite-derived immune-suppressive factors on the cellular innate immune and autoimmune responses of Drosophila melanogaster
[Articolo su rivista]
Nappi, A. J.; Vass, E.; Malagoli, Davide; D., Carton
abstract
Immune-suppressive factors (ISFs) introduced into larvae of Drosophila melanogaster during infection by virulent endoparasitic wasps effectively block the innate immune response mediated by blood cells (hemocytes) but have little influenceon the autoimmune response made by a tumor strain in which the blood cells manifest a similar response but instead target anddestroy endogenous tissues. Quantitative hemocyte analyses indicate that ISFs interfere with the immune effector responsesdownstream of nonself recognition, hemocyte activation and differentiation, because these responses were manifested by tumorhosts, in which the parasitoids developed. The data suggest that once activated to encapsulate aberrant tissues, the target specificityof the autoimmune-activated hemocytes, and the genetic program underlying tumor formation, cannot be blocked by parasitoidderivedISFs, which effectively inhibit identical hemocyte-mediated responses during parasitization.
2004
- Yessotoxin affects fMLP-induced cell shape changes in Mytilus galloprovincialis immunocytes
[Articolo su rivista]
Malagoli, Davide; Ottaviani, Enzo
abstract
Using computer-assisted microscopic image analysis, we have found that algal yessotoxin (YTX) affects the immune response of Mytilus galloprovincialis. Indeed, YTX increases immunocyte cell motility through the involvement of both extracellular Ca2+ and CAMP. but not through protein kinase A, protein kinase C or phosphoinositide 3-kinase. Alone, however, the toxin does not induce any effect, as its action on cell motility is observed only after addition of the chemotactic substance N-formyl-Meth-Leu-Phe (fMLP). fMLP is known to induce cellular changes via both the phosphatidylinositol and CAMP pathways and, from this scenario, we can surmise that Ca2+ and CAMP concentrations rise sufficiently in fMLP-activated immunocytes to reveal YTX action. One possible explanation is that the toxin increases fMLP-mediated cell activation by intervening in L-type Ca2+-channel opening through a cAMP-dependent/PKA-independent pathway.
2003
- Effects of 50 hz magnetic fields on fMLP-induced shape changes in invertebrate immunocytes: The role of calcium ion channels
[Articolo su rivista]
Gobba, Fabriziomaria; Malagoli, Davide; Ottaviani, Enzo
abstract
Plasma membrane Ca2+ channels in immunocytes from the mussel Mytilus galloprovincialis exposed to 50 Hz sine wave magnetic fields (MFs) of various strengths were studied. At levels of 300 muT and above, MFs reduce shape changes in immunocytes induced by the chemotactic substance N-formyl-Meth-Leu-Phe, and this effect involves L-type Ca2+ channels. Upon the addition of the Ca2+ blocker verapamil to molluscan immunocytes exposed to MFs results in a synergistic cytotoxic action, while in the presence of the Ca2+ opener SDZ-202, 791, a reactivation of the cells is observed. This suggests that, as previously reported for potassium channels, the damage to Ca2+ channels induced by short exposure to MF at appropriate intensities is not permanent.
2003
- Effects of 50-Hz magnetic fields on the signalling pathways of fMLP-induced shape changes in invertebrate immunocytes: the activation of an alternative stress pathway
[Articolo su rivista]
Malagoli, Davide; Gobba, Fabriziomaria; Ottaviani, Enzo
abstract
N-formyl-Meth-Leu-Phe (fMLP)-induced immunocyte shape changes in the mussel Mytilus galloprovincialis through both the phosphatidylinositol and the cAMP pathways are studied. Fifteen- and thirty-minute exposures of mussels to 50-Hz magnetic fields (MFs) at intensities of 300 and 400 muT do not provoke permanent cell damage, since immunocytes maintain the capacity to respond to fMLP. This avoidance of external insult seems to be achieved through the activation of a stress pathway which is not functionally detectable in nonexposed animals and which involves mitogen activated protein (MAP) kinase members. This phenomenon is clearly evident at 400 muT. Contemporaneously, a different expression of Jun transcriptional regulatory proteins is also found. (C) 2002 Elsevier Science B.V. All rights reserved.
2002
- 50 Hz magnetic fields of varying flux intensity affect cell shape changes in invertebrate immunocytes: the role of potassium ion channels
[Articolo su rivista]
Ottaviani, Enzo; Malagoli, Davide; A., Ferrari; Tagliazucchi, Davide; Conte, Angela; Gobba, Fabriziomaria
abstract
The effect induced by exposure to 50 Hz magnetic fields (MFs) in immunocytes from the mussel Mytilus galloprovincialis is evaluated. The whole animal was exposed for 15 and 30 min to MF intensities ranging from 200 to 1,000 microT. The changes in the cellular shape of immunocytes, expressed as shape factor (SF), were studied at different times after addition of the chemotacting substance N-formyl-Meth-Leu-Phe (fMLP). Results show that MFs provoke differing delays in fMLP-induced cellular shape changes: 200 microT are ineffective, while levels from 300 microT upwards cause a significant increase in immunocyte SF values compared to controls. Reactivation of the cells is possible up to an intensity of 600 microT. The use of PCO 400, an opener of ATP-sensitive K+ channels, shows that potassium channels are involved in the effect of MFs on M. galloprovincialis immunocytes.
2002
- Bcl-2 does not control programmed cell death in the IPLB-LdFB cell line from the insect Lymantria dispar
[Articolo su rivista]
Ottaviani, E.; Malagoli, D.
abstract
In the insect Lymantria dispar cell line IPLB-LdFB the presence of a Bcl-2-like molecule has been demonstrated. The Western blot analysis performed on the cells incubated with 2-deoxy-D-ribose (dRib), an apoptotic inducer, revealed that, in comparison with the control, the Bcl-2 expression was unaffected. Furthermore, incubation of the insect cells with an anti-Bcl-2 polyclonal antibody inhibited the apoptotic effect induced by dRib, and provoked mitochondrial membrane depolarization without any apoptotic phenomena. Similar behaviour was observed using the K+ ionophore valinomycin. From these findings, we hypothesize that the L. dispar Bcl-2-like protein is essential for maintenance of the mitochondrial membrane potential, but not, as usually thought, for the regulation of programmed cell death. (C) 2002 Elsevier Science Ltd. All rights reserved.
2002
- Cloning and characterisation of a procorticotrophin-releasing hormone in the IZD-MB-0503 immunocyte line from the insect Mamestra brassicae
[Articolo su rivista]
Malagoli, Davide; Mandrioli, Mauro; Ottaviani, Enzo
abstract
The cloning and characterisation of a procorticotrophin-releasing hormone (proCRH) and the related CRH fragment in the IZD-MB-0503 cell line from the leptidopteran Mamestra brassicae were performed. PCR amplification of the genomic DNA reveals a fragment of 276 bp, while inverse PCR shows the presence of a gene consisting of 1153 bp. The comparison of the insect genomic proCRH gene with proCRH cDNA obtained by RACE shows the presence of three introns. There was a 61% identity with the corresponding coding sequence in Tilapia mossambica, and a 65.2% identity with the human proCRH coding sequence. (C) 2002 Elsevier Science Inc. All rights reserved.
2002
- Protein kinases mediate nitric oxide-induced apoptosis in the insect cell line IPLB-LdFB
[Articolo su rivista]
Malagoli, Davide; Conte, Angela; Ottaviani, Enzo
abstract
The involvement of protein kinases (PKA, PKC and PKB) in nitric oxide (NO)-induced apoptosis with sodium nitroprusside plus N-acetyl-L-cysteine in the IPLB-LdFB cell line from the insect Lymantria dispar was investigated. The presence of protein kinase-like molecules was demonstrated by Western blot analysis. The role of the kinases in programmed cell death was analysed in cytofluorimetric experiments by incubating the insect cells with H-89 (a specific inhibitor of PKA), calphostin C (an inhibitor of PKC) or wortmannin (an inhibitor of phosphatidylinositol 3-kinase). The results show that PKA is correlated with the induction and PKC and PKB with the prevention of NO-induced insect cell death. Moreover, NO-induced apoptosis involves the release of cytochrome c.
2001
- An anti-Bcl-2 antibody prevents 2-deoxy-D-ribose-induced apoptosis in the IPLB-LdFB insect cell line
[Articolo su rivista]
D., Barbieri; Malagoli, Davide; B., Cuoghi; Ottaviani, Enzo
abstract
Confocal microscopy reveals that the anti-Bcl-2 antibody (pAb) is able to diffuse across the plasma membrane of the fat body cell line IPLB-LdFB from the insect Lymantria dispar demonstrating the presence of Bcl-2-like molecules in the cytoplasm. Immunoperoxidase procedure confirms the cellular localization. Furthermore, an immunoprecipitation corresponding to a molecular weight of 29 KDa is observed with western blot analysis using the anti-Bcl-2 pAb. Cytofluorimetric experiments show that anti-Bcl-2 pAb counteracts 2-deoxy-D-ribose-induced apoptosis and provokes morphological changes in the insect cell line, i.e. a reduction in cell size, the disappearance of the vacuola and changes in shape. At the same time, the antibody provokes mitochondrial membrane depolarization, and N-acetyl-L-cysteine is unable to reconstitute the physiological conditions. The present findings suggest that Bcl-2-like proteins play a main role in maintaining of the integrity of cellular components, e.g, mitochondria, rather than in controlling programmed cell death.
2001
- Involvement of PI 3-kinase, PKA and PKC in PDGF- and TGF-β-mediated prevention of 2-deoxy-D-ribose-induced apoptosis in the insect cell line, IPLB-LdFB
[Articolo su rivista]
Ottaviani, E.; Barbieri, D.; Malagoli, D.; Kletsas, D.
abstract
Activation of phosphatidylinositol (PI) 3-kinase, protein kinase A (PKA) and protein kinase C (PKC) is associated with the survival effect elicited by PDGF-AB and TGF-β1 against the apoptotic inducer 2-deoxy-D-ribose (dRib) in the fat body cell line, IPLB-LdFB, from the insect Lymantria dispar. dRib induces apoptosis and provokes mitochondrial membrane depolarization (MMD). The antioxidant N-acetyl-L-cysteine annuls only the first effect. These findings suggest that apoptosis and MMD are provoked by two different mechanisms, and that dRib induces apoptosis by oxidative stress. © 2001 Academic Press.
2001
- Nitric oxide induces apoptosis in the fat body cell line IPLB-LdFB from the insect Lymantria dispar
[Articolo su rivista]
Ottaviani, Enzo; D., Barbieri; Malagoli, Davide; Franchini, Antonella
abstract
The presence of immunoreactive inducible nitric oxide synthase molecules (ir-iNOS) is demonstrated in the Lymantria dispar IPLB-LdFB cell lint. The maximum ir-iNOS inducibility is observed 18 h after incubation with sodium nitroprusside (SNP). The increase in NO provoked by SNP in turn induces apoptosis. However, this phenomenon is observed only after 48 h. The NOS-inhibitors N-omega-nitro-L-arginine methyl ester (L-NAME) and N-[3-(aminomethyl)-benzyl]acetamide (1400W) were both unable to block the SNP-induced apoptosis at all the concentrations used. Incubation with SNP plus N-acetyl-L-cysteine (NAC) further augmented the percentage of cell death with respect to SNP used alone, and this process is seen earlier, i.e. after 24 h. Moreover, the induction of apoptosis in the presence of NAC is time- and concentration-dependent. The high percentage of cell death with SNP + NAC suggests that NAC forms S-nitrosothiols with NO, resulting in an increase in the bioavailability of NO. In conclusion, these findings show the existence of a close relationship between mammalian and invertebrate cells with regards to SNP and NAC induction and the related NO response. (C) 2001 Elsevier Science Inc. All rights reserved.
2000
- Immunomodulation by recombinant human interleukin-8 and its signal transduction pathways in invertebrate hemocytes
[Articolo su rivista]
Ottaviani, Enzo; Franchini, Antonella; Malagoli, Davide; Genedani, Susanna
abstract
We report the presence of interleukin (IL)-8-immunoreactive molecules in hemocytes from the mollusc Mytilus galloprovincialis. Functional studies demonstrate that recombinant human (rh)IL-8 provokes conformational changes, induces chemotaxis, and increases bacterial phagocytic activity in hemocytes. rhIL-8 induces cell shape changes via protein kinase A and C pathways. These morphological changes are followed by reorganization of the actin microfilaments. The findings suggest that, as previously reported for other cytokines, IL-8 is well conserved and deeply involved in immune functions from invertebrates to mammals.
2000
- Synergistic role of cAMP and IP3 in corticotropin-releasing hormone-induced cell shape changes in invertebrate immunocytes
[Articolo su rivista]
Malagoli, Davide; Franchini, Antonella; Ottaviani, Enzo
abstract
Immunocytes from the mollusc galloprovincialis express corticotropin-releasing hormone (CRH) receptor subtype (CRH-R1 and CRH-R2)-like mRNAs. Using computer-assisted microscopic image analysis, we have found that exogenous CRH provokes changes in the cellular shape of immunocytes, and that this response is extracellular Ca2+-dependent. The various inhibitors of transduction signaling pathways, i.e. suramin sodium, 2',5' -dideoxyadenosine: neomycin sulfate, calphostin C, H-89, and wortmannin, completely or partially inhibit these changes. The present findings demonstrate that PKA, PKC, and PKB/Akt are involved in CRH-induced cell shape changes in immunocytes, and that the cellular effect of CRH needs the synergistic action of the two second messengers, cAMP and IP3. (C) 2000 Elsevier Science Inc. All rights reserved.
1999
- Platelet-derived growth factor and transforming growth factor-β induce shape changes in invertebrate immunocytes via multiple signalling pathways and provoke the expression of Fos-, Jun- and SMAD-family members
[Articolo su rivista]
Ottaviani, E.; Malagoli, D.; Kletsas, D.
abstract
Recently, we have found that platelet-derived growth factor (PDGF)-AB and transforming growth factor (TGF)-β1 induce shape changes in the immunocytes of the mollusc Mytilus galloprovincialis. Here we report that calphostin C and H-89, which specifically inhibit PKC and PKA, respectively, completely or in part inhibit the above effect. These data indicate the involvement of both pathways in growth factor-induced conformational changes in molluscan immunocytes. Furthermore, we show the presence of immunoreactive molecules for the Fos B, Jun D and Smad4 transcription factors. Stimulation by PDGF-AB or TGF-β1 up-regulate the intranuclear levels of these factors. Copyright (C) 1999 Elsevier Science Inc.