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SummaryHigh-intensity femtosecond pulses from an X-ray free-electron laser enable pump probe experiments for investigating electronic and nuclear changes during light-induced reactions. On time scales ranging from femtoseconds to milliseconds and for a variety of biological systems, time-resolved serial femtosecond crystallography (TR-SFX) has provided detailed structural data for light-induced isomerization, breakage or formation of chemical bonds and electron transfer1. However, all ultra-fast TR-SFX studies to date have employed such high pump laser energies that several photons were nominally absorbed per chromophore2-14. As multiphoton absorption may force the protein response into nonphysiological pathways, it is of great concern15whether this experimental approach16allows valid inferences to be drawn vis-à-vis biologically relevant single-photon-induced reactions17. Here we describe ultrafast pump-probe SFX experiments on photodissociation of carboxymyoglobin, showing that different pump laser fluences yield markedly different results. In particular, the dynamics of structural changes and observed indicators of the mechanistically important coherent oscillations of the Fe-CO bond distance (predicted by recent quantum wavepacket dynamics15) are seen to depend strongly on pump laser energy. Our results confirm both the feasibility and necessity of performing TR-SFX pump probe experiments in the linear photoexcitation regime. We consider this to be a starting point for reassessing design and interpretation of ultrafast TR-SFX pump probe experiments16such that biologically relevant insight emerges.

Regenerative capability in anthozoans is an important adaptive strategy for their survival to environmental disturbance of natural and anthropogenic origin such as predation or anchoring, that can cause injuries or removal of entire parts of the animal body, and it can be also considered indirectly a further tool of innate immune system. In the context of “self”-“non self” recognition, is significant the interaction with the endosymbiont of the genus Symbiodinium and the recognition of pathogens and foreign agents capable of invading the injured tissues. From these premises and the growing stressors that can cause injuries, it is significant to understand how species respond to physical damage and how they manage to recover and regenerate compromised tissues. Our research team, studied in the Mediterranean anthozoan Anemonia viridis (Forsskål, 1775), the natural seasonal variability of its morphology and enzymatic biomarkers involved in inflammatory process, the immune system response following injection of molecules varied in type and dimension. In particular, after the infection of two pathogenic bacteria Escherichia coli and Vibrio alginolyticus a particular and strong reaction was observed. These previous knowledge allowed us to examine the activity of enzymes such as proteases (SDS-PAGE on gelatin and fibrinogen substrate), peroxidase and alkaline phosphatase as biomarkers traditionally involved in wound healing event and in the rearrangement of extracellular matrix. The regenerative process, in this Mediterranean species of anthozoan, was analyzed by subjecting groups of animals to differential tentacle cuts (n = 10, 20, 30) and observing and estimating the regenerative potential after 7, 14 and 21 days from cutting. A morphological and histological observation of the tentacular regrowth event was conducted in addition to the evaluation of the expression of proliferating cell nuclear antigen (PCNA) using the immunoblotting technique. The future goal is to increase the knowledge of the processes that trigger the immune response in inflammation and regeneration, for the considerable interest in basic sciences and for the transferability of results in biotechnological field

The diversification of anthozoan toxins played an important role in the ability to colonize various ecological niches. In this study we evaluated the hemolytic activity of HPLC separated fraction of tentacle extracts of sea anemone Actinia equina. Toxic components from acid tissue tentacle extracts were investigated by size exclusion and reverse phase HPLC to characterize cytolytic molecules. A novel low molecular weight active fraction was sequenced by MALDI TOF analysis and a protein correspondent to 5.4 kDa Sodium channel neurotoxin (Ae1) from A. equina was identified. Synthetic Ae1 was assayed and it showed an hemolytic activity against mammalian erythrocytes in a dose dependent manner. Cytolytic activity in addition to neuro-inhibitory function could be a further property of Ae1 toxin repertoire., Invertebrate Survival Journal, Vol 13 No 1 (2016)

Gilthead seabream (Sparus aurata) is the only species of the Sparidae family currently bred on a large scale. As one of the most important species on Mediterranean aquaculture. For this main reason, the knowledge of its immune system and their alterations it is crucial to avoid economic losses worldwide. Fish are in intimate contact with their environment and defending itself by a complex system of innate defence mechanisms, that is divided into physical barriers, cellular and humoral components. The skin as a physical barrier seems to be central to protection from pathogens such as Photobacterium damselae ssp. piscicida. Probiotics are microorganisms (usually bacteria) that promote different benefits on host. The aim of this work was to evaluate the in vitro capacity of Shewanella putrefaciens also known as Pdp11 (a probiotic isolated from skin of gilthead seabream) to improve the skin immune defence against Photobacterium damselae. Thus, we analyzed by qPCR the expression profile of different cytokines in seabream skin explants, in response to an exposure (of 2 and 4 hours) to the probiotic Pdp11 and the pathogen Photobacterium damselae. Our results showed that the different cytokines are modulated by both the pathogen (P. Damselae) and the probiotic (Pdp11), which could, at least in in vitro level, improve the gilthead seabream mucosal health. Further studies are neccesary to unravel if the present findings in vitro can be confirmed through in vivo studies

Immunomodulation and physiological responses of Mytilus galloprovincialis as bioindicators of environmental change Oxygenation level temperature increases and changes in food availability are predicted to occur in the future. In such scenario, a global climate change (GCC), there is growing concern for the health status of wild and farmed organisms. Bivalve molluscs, are important components of coastal marine ecosystems, and as sedentary and filter feeders, are good bioindicators of environmental conditions. The ability of organisms to maintain the immunosurveillance unaltered under adverse environmental conditions may enhance theirs survival capability. Only a few studies have investigated the effects of changing environmental parameters on the mussels immunity. In the present study, the effects of different food concentration, temperature and oxygenation treatments were evaluated on immune parameters of Mytilus galloprovincialis detected on digestive gland and hemocytes. Bivalves were exposed to experimental conditions by increasing of six food treatment, to three different temperatures under conditions of normoxia and anoxia. The multifactorial analysis applied to the responses of the immune variables has showed a direct dependence of various enzymes production by temperature and food concentration. The stability of the lysosomal membrane was altered under conditions of thermal stress and food changing. The protein concentration of the lysate of hemocytes instead was most affected by the lack of adequate oxygenation. In addition, a correlation was carried out between mussels immunological effectors and physiological responses as clearance rate, measured by the removal of suspended particles from water flowing through experimental chambers, food absorption efficiency and rates of oxygen consumption by individual mussels. Overall, information summarized in the present study indicated that climate changes can affect hemocyte and enzymatic functionality and the immune responses of this bivalve could be used as good environmental biomarkers.

Variation of environmental condition and diet act on immune parameters of Mytilus galloprovincialis The knowledge of the immunity mechanisms as environmental indicators and their alterations in the case of physical stress can be of fundamental importance in the environmental management programs. Recently has been shown that environmental factors affect immune responses in some species of bivalves. In this study we assessed different enzymatic activities from digestive gland of M. galloprovincialis such esterase, phosphatase and phenoloxidase (PO), involved in digestive inflammatory, detoxification and melanization processes. Particularly, esterases catalyze the hydrolysis reaction of the ester bond. Phosphatases modulates the removal of phosphate groups by producing phosphoric acid from esters and participate in the metabolism of sugars, nucleotides and phospholipids. The melanization cascade, in which phenoloxidase is the terminal enzyme, plays a key role in recognition of microbial infections in molluscs. It was also evaluated, as potential biomarker, the lysosomal membrane stability through neutral red assay on hemocytes taken from the posterior adductor muscle. Specimens were maintained under conditions of normoxia and anoxia and they were subjected to various food amount and different temperatures. 102 The results showed that the enzymatic activities of esterases, phosphatases and PO are higher during treatments with lowest temperatures and food amount. Moreover, the production of PO is higher in the conditions of anoxia. The lower values of enzymatic production have been detected under the levels of temperature, oxygen availability and food different than the optimum conditions for the mussels life cycle. During the normoxic treatment, the stability of the lysosomal membrane is highest at the average values of temperature and food concentration. The lowest values, instead, were measured at a temperature of 12 °C and 28 °C in anoxic conditions

Light absorption can trigger biologically relevant protein conformational changes. The light-induced structural rearrangement at the level of a photoexcited chromophore is known to occur in the femtosecond timescale and is expected to propagate through the protein as a quake-like intramolecular motion. Here we report direct experimental evidence of such ‘proteinquake’ observed in myoglobin through femtosecond X-ray solution scattering measurements performed at the Linac Coherent Light Source X-ray free-electron laser. An ultrafast increase of myoglobin radius of gyration occurs within 1 picosecond and is followed by a delayed protein expansion. As the system approaches equilibrium it undergoes damped oscillations with a ~3.6-picosecond time period. Our results unambiguously show how initially localized chemical changes can propagate at the level of the global protein conformation in the picosecond timescale., Localized chemical events such as the breakage of a bond between a protein and a ligand may trigger a global protein conformational change. Here, the authors use an X-ray free-electron laser to track the motion of myoglobin in response to photoinduced ligand release, and observe a picosecond proteinquake.

Progetto selezionato ADI DESIGN INDEX 2015 e candidato al compasso d'oro 2016. Corporate image/identity e supporti di wayfinding per struttura ospedaliera a Gela. Il progetto comunicativo si innesta in un ampio programma di trasformazione della clinica che ha interessato la riorganizzazione interna dei servizi – la Clinica, gli Ambulatori, la Diagnostica e la nuova Residenza Sanitaria Assistita (RSA) – l’ammodernamento dell’edificio, l’efficientamento delle modalità operative e dei protocolli per l’erogazione dei servizi, oltre che al normale e continuo aggiornamento del personale e all’adeguamento delle attrezzature. All’interno di questo processo dinamico, che mira al consolidamento di un nuovo profilo di “ospedale senza dolore, la strategia di identità visiva coordinata elabora una lettura del brand specifico (a partire dal naming), del concetto stesso di luogo per la cura e dei supporti a servizio del personale e dell’utenza. L’uso di strumenti digitali come il QR-CODE, applicato a tutti i prodotti cartacei progettati, riposiziona ogni piccolo oggetto in una famiglia di oggetti vasta e coerente che uniforma la segnaletica, le divise del personale, le insegne, il nuovo logotipo e l’intera immagine dell’ospedale attraverso il nuovo sito web www.santabarbarahospital.it. Project selected for ADI DESIGN INDEX 2015 and candidate for the Compasso d'Oro in 2016. Corporate identity and media wayfinding for the hospital in Gela. The communication design is part of an extensive program of the clinic’s transformation that involved the internal re-organization of services, modernization of the building, improving the efficiency of the operating procedures and protocols for the delivery of services in addition to the normal and continuous update of staff and the upgrading of equipment. Within this dynamic process, which aims at consolidating a new profile of “pain-free hospital”, the strategy of coordinated visual identity develops a reading of the specific brand (starting with the naming), the same concept of a place for care and attention and support for service personnel and users. The use of digital tools such as QR CODEs, applied to all the designed paper products, repositions every small object in a wide and coherent range of items that applies a level of conformity to all signs, the staff uniforms, the internal and external communication, the new logo and the entire image of the hospital through the web site www.santabarbarahospital.it.

The phylum of Cnidaria is one of the first branches in the tree of animal life to provide crucial insights on the evolution of immunity. Cnidarians are diblastic aquatic animals with radial symmetry and they are the simplest multicellular organisms that have reached the level of tissue organization. The renewed interest in the study of immunity in Cnidaria has led to additional information to the scenario of the first stages of immunity evolution revealing the cellular processes involved in symbiosis, in the regulation of homeostasis and in the fight against infections. We investigated the inflammatory response in Cnidarian following injection of various substances different in type and dimension, and observed clear, strong and specific reactions especially after injection of bacteria. The enzymes evaluation (protease, phosphatase and esterase), showing how the injection of different bacterial strains alters the expression of these enzymes suggesting a correlation between the appearance of the inflammatory reaction and the modification of enzymatic activities. The Cnidaria phylum has evolved using biotoxins as defense or predation mechanisms for ensure survival in hostile and competitive environments such as the seas and oceans indeed the tissues and the mucus produced by cnidarians are involved in immune defense and contain a large variety of toxins such enzymes, potent pore forming toxins, and neurotoxins. They could also take advantage of the multi-functionality of some of their toxins. The bioactive molecules were characterized and purified by biological assays, acid extraction, HPLC purifications, mass spectroscopy and peptide synthesis. Here, we show the cnidarian bioactive molecules as antimicrobial peptides and enzymes in order to draw important applications in fields ranging from pharmacology to cultural heritage.

The Cnidaria phylum includes organisms that are among the most poisonous animals. The exact composition of cnidarian bioactive molecules is not known in detail, but little is known on the cells that produce the toxins. Here we have shown that the presence of cytolysins is not exclusive of nematocysts. A plaque-forming assay was carried out with cell populations extracted from the percoled body fluid showed for the first time that anthozoan granulocytes are able to form plaque of lysis. We have partitioned the total population of free cells into three distinct discrete bands by discontinuous Percoll gradient, and we have identified six small different types cells: morular granulocytes; cells with large or small peripherical granules, granulocytes with irregular shape containing blue and red granules, cells showing one fine red granule of uniform size and, finally, cells with elongated shape and small dispersed granules. Cell lysate of each cellular band resulted cytolytic toward different erythrocytes types. SDS page analysis of the lysate cell fraction showed a predominant of 20 kDa that corresponds to the weight of the cytolytic equinatoxin. The nature of equinatoxins-related activity was demonstrated by inhibition experiments using bovine sphingomyelin.

The paper concerns the seismic behavior of precast concrete industrial buildings with an elevation equipped with seismic isolators. It’s examined a building with rectangular plan of m 40x 50. The structure is constituted of columns of square cross-section of 0.6 m, with continuum beams foundation. The desk is made with T section beams for each frame connected by Gerber joints in the main direction and they support precast pre-tensioned tiles with  shape. A plate of concrete is casted of the wing tiles for obtain a connected rigid deck. At the top of the pillars will be placed rubber bearings having high dissipation capacity (HDRB). The dynamic behavior of the structure under recorded real strong accelerograms is examined. In this work are considered Friuli, Loma Prieta, Northridge and Kobe accelerograms, dynamic incremental analysis are conducted in order to obtain the response of structure. The concrete behavior is assumed with Mander law for the pillars, while the beams have a linear elastic behavior. For the isolators a linearized constitutive law, using equivalent stiffness, is assumed. The design criteria of isolated precast systems are highlight, in detail the use of isolators permit to reduce the dimensions and the reinforcing bars amount of pillars. It is need to connect the precast beams of the deck for obtain a rigid deck motion.

in tedesco del Poster Multiple Wesen, esposto al Simposio internazionale „Lunar Base. Bauen fuer ein Leben auf dem Mond” tenuto a Kaiserslautern il 12-13 Maggio 2009. Was baut man auf dem Mond? Wie baut der Erdmensch auf dem Mond? Wofür wird gebaut? Für wem wird gebaut? Welche Nutzungen, wie groß, welche Materialien? Man kann weiter fragen,... Sind dieses überhaupt die richtige Fragen? Kann man überhaupt vom Bauen auf dem Mond sprechen? Oder geht es darum, was der Mensch auf den Mond mit bringt, aus seiner Kenntniss, seinen Vorstellungen, seinen Träumen, seiner Erfahrung: was bringt er von dem Reich des Bekannten (der Erde) in das Reich des nur Vorstellbaren (dem Mond). Also: sich mit dem Mond zu beschäftigen - ausgenommen für die sehr wenigen, die schon dort waren – heißt, sich mit sich selbst zu beschäftigen, mit dem eigenen Wissen und dem Bild vom Mond. Alle Voraussetzungen, die einem irdischen architektonischen Entwurf zu Grunde liegen, fehlen; zu diesem Zeitpunkt kann man sich arbiträr Rahmenbedingungen geben: (Größe, wo, welche Nutzung, welche Lebensdauer, welche Materialien) so zu sagen ein Mondprojekt in einem Erdprojekt zu entwickeln, dadurch schnell eine „abgesicherte“, unangreifbare Lösung zu erlangen. Aber macht dieses Sinn? Bedeutet dies, die Spannung und die Erforschung des Neuen sowie das kreative Potential örtlicher Schwierigkeiten im Vorfeld auszuschließen? Wir schlagen vor, anstatt auf hiesige Fragen irdische Antworten zu formulieren, Strukturen zum Mond zu bringen; Raumstrukturen, die durch mentale Konzepte begründet sind, welche die vielen Fragen - auch welche die wir jetzt nicht zu fragen gewagt haben - aufnehmen können. Diese Strukturen, aber auch kleine räumliche Gebilde, helfen dem Vorstellungsvermögen der Forscher, Ingenieure, Wissenschaftler und alle Mondinteressenten. Neutrale Strukturen, ohne jetzt genau alle physische Eigenschaften festlegen zu wollen, sollen Raum werden können für Wohnen, Arbeiten, Forschen, Landen, Binden, Schützen, Aufbewahren, usw. - abhängig von den Fragen, die sich zu gegebener Zeit formulieren lassen und ständig ändern werden. Wir denken an sehr flexible Strukturen, die viel aufnehmen und aushalten können, sich sehr schnell transformieren lassen, also multiple Wesen, die zunächst die Vorstellung eines belebten Mondes anregen sollen. Dieser Strukturenkatalog soll dem Wissenschaftler zur Verfügung stehen, um das Mitdenken seiner räumlichen Bedürfnisse zu stützen. Comunicato stampa Contributo degli studenti di Architettura al Simposio “Lunar Base - Costruire per una vita sulla Luna” A Kaiserslautern in Germania è in corso (12-13-maggio) il Simposio “Lunarbase - Bauen fuer ein Leben auf dem Mond” (www.lunar-base.net) organizzato dalla DLR (Agenzia aerospaziale tedesca) e dalla TU Kaiserslautern. Scienziati, ingegneri, architetti e designer di tutto il mondo presentano i loro piani e le loro visioni per una possibile vita sulla Luna. Alcune nazioni tra le quali la Cina, la Russia e gli Stati Uniti stanno elaborando piani concreti per la costruzione di stazioni lunari come basi per missioni equipaggiate su Marte, con un orizzonte temporale intorno al 2020. La proposta degli studenti dei Laboratori di I anno di Progettazione architettonica, tenuti dal Prof. Gaetano Licata con la collaborazione del dottorando in Design Michele Cammarata nel nuovo corso di Laurea triennale in Scienze dell’Architettura (SdA) e nel Corso di Laurea quinquennale (4/s), è stata selezionata dal comitato organizzatore che ne ha richiesto l’elaborazione sotto forma di un Poster (A0) attualmente esposto durante il Simposio. Si tratta di un montaggio di modelli tridimensionali elaborati dagli studenti e denominati “multiple Wesen”. Questi modelli, in quanto concetti di architettura assolutamente astratti, si prestano a contribuire a quel bagaglio di conoscenze, visioni, sogni e immaginazione, che chiunque voglia “confrontarsi” con una realtà solo immaginabile come quella della Luna, ha bisogno di “portare con sé” dalla Terra. Concetti mentali di spazi possibili – “Raumstrukturen” - a sostegno degli scienziati, per poter continuare a immaginare insieme sempre più concretamene la vita e la ricerca direttamente sulla Luna. Pa, 13.05.2009 (gaetano.licata@unipa.it)

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.

A theoretical study of the ultrafast electron kinetics in solid SiO2, irradiated with a femtosecond X-ray laser pulse, is presented. A Monte Carlo code for event-by-event simulations of individual particles is applied to model the electron kinetics within the irradiated SiO2 bulk. The simulation includes photoionization, elastic and inelastic scatterings of electrons, Auger decays of core holes, and electron-hole recombination via exciton self-trapping mechanism. Transient electron density is followed, at different photon energies and pulse durations. The change of the optical properties (reflectance, transmittance) of the material is estimated with the help of the Drude model. The analysis of the results allows us to conclude that within the X-ray excited dielectric, the holes in the valence band give the predominant contribution to the transient changes of optical properties within the material. The increase rate of the free electron density is limited by the duration of secondary electron cascading.

Several diagnostic genetic markers were identified in Pomatoschistus marmoratus and P. tortonesei using polyacrylamide gel electrophoresis (PAGE) of allozymes. Twenty-one loci were resolved, including the electrophoretic pattern of muscle proteins. The MDH*, PGM- 1,2*, EST-1,2*, FUM* and PGI-2* loci exhibited different alleles which were fixed for the two species being analysed. Genetic distance, as calculated by Nei’s index, showed a value of 0·413. Environmental hypersalinity, could have influenced the geographical distribution of P. tortonesei.

The cytotoxic activity against rabbit erythrocytes (RE) and human K562 tumor cells by Styela plicata hemocytes was significantly related to the phenoloxidase (PO) which converts phenols to quinone and initiates the melanogenic pathway. The effector hemocyte population, separated in a Percoll density gradient band, enriched in a granulocyte type named "morula cells", was examined with RE in a hemocyte cytotoxic assay and plaque forming cell assay. Inhibition experiments with the copper chelating agents 1-phenyl-2-thiourea and tropolone, the substrate analogue sodium benzoate and sodium ascorbate support the notion that hemocyte cytotoxic activity is a PO-dependent mechanism. Treatments of hemocytes with the antioxidant enzymes, superoxide dismutase and catalase rule out oxy radicals produced by the melanogenic process as responsible of erythrolysis. Such a result suggests that quinone compounds derived from the melanogenic pathway might be the cytotoxic molecules. The PO-dependent anti-RE activity was also shown in a plaque forming assay in which "morula cells", containing polyphenols and PO, were identified as cytotoxic.