Dereplicazione di "miceli sterili" mediante tecniche di fingerprinting molecolare

Il significato principale del termine dereplicazione fa riferimento alla biochimica: indica il processo con cui componenti facenti parte di una sostanza più complessa, vengono analizzati per riconoscere ed eliminare le sostanze attive già studiate. Tale concetto è stato utilizzato anche per semplificare l’analisi di comunità di organismi campionati, al fine di identificare individui uguali non distinguibili da un punto di vista morfologico o strutturale. In questo lavoro la dereplicazione è stata applicata allo studio di miceli sterili, mediante l’analisi di fingerprinting molecolari ottenuti amplificando il DNA genomico con il minisatellite M13. La tecnica del fingerprinting molecolare utilizzando l’M13 come sonda è stata utilizzata per differenziare funghi filamentosi a livello di genere e di specie consentendo, inoltre, l’individuazione di cloni di un singolo isolato attraverso fingerprinting identici. Non vi sono notizie di lavori di dereplicazione di funghi filamentosi con l’utilizzo del minisatellite M13 come primer in esperimenti di PCR. I miceli sterili sono definiti “funghi che mancano della produzione di spore di ogni tipo”. Poichè la sistematica fungina è basata sulla morfologia e sulla strutture delle spore, la loro completa assenza rende i miceli sterili virtualmente non identificabili, né a livello di famiglia, né di genere o specie. Tuttavia, l’approfondimento delle conoscenze biologiche, fisiologiche e tassonomiche su tali funghi, spesso trascurati per le notevoli difficoltà che si incontrano nel loro studio, risulta di estremo interesse viste le loro potenzialità applicative come promotori della crescita delle piante o agenti di biocontrollo. In questo lavoro sono stati analizzati 99 miceli sterili, isolati da paglia interrata in due terreni (sabbioso e argilloso) con una precedente storia di coltivazione a frumento. L’isolamento è stato effettuato nel corso di una prova per la competizione per la paglia da parte di funghi filamentosi in presenza di Deossinivalenolo. Il DNA estratto è stato amplificato con il primer M13 e i prodotti di PCR sono stati separati mediante elettroforesi su gel di agarosio ottenendo fingerprinting con un numero di bande compreso tra 4 e 9, negli isolati campionati dal terreno sabbioso, e tra 4 e 13, negli isolati provenienti dal terreno argilloso. Per entrambi i terreni, i dati ottenuti dall’analisi delle bande di ciascun isolato sterile, sono stati utilizzati per costruire una matrice binaria, in cui 1 indica la presenza e 0 l’assenza della banda di ogni isolato. E’ stata quindi elaborata, con il programma Past, una matrice di somiglianza utilizzando il coefficiente Jaccard, dalla quale è stato poi costruito un dendogramma con il metodo UPGMA. L’analisi cluster dei 42 miceli sterili provenienti dal terreno sabbioso, ha evidenziato la presenza di 6 gruppi di isolati con fingerprinting identici e una similarità del 100%. L’analisi ha consentito, inoltre, di individuare un cluster di 11 isolati con profili elettroforetici molto simili (similarità 80%). Per il terreno argilloso, l’analisi cluster dei 57 miceli sterili ha mostrato la presenza di 13 gruppi di isolati con fingerprinting identici e una similarità del 100%. Dal dendrogramma è emersa, inoltre, la presenza di due cluster rispettivamente di 5 e 6 isolati con fingerprinting molto simili e percentuali di similarità pari a 75 e 88. I rimanenti isolati si sono raggruppati in numerosi cluster con una somiglianza inferiore al 60%. La dereplicazione dei miceli sterili è stata, quindi, effettuata in entrambi i tipi di terreno. Nel terreno sabbioso sono stati dereplicati 11 miceli sterili sui 42 analizzati (26%) e nel terreno argilloso ne sono stati dereplicati 20 sui 57 inizialmente saggiati (35%). I risultati ottenuti nel presente lavoro possono essere considerati incoraggianti per sviluppare la metodologia di dereplica di miceli sterili mediante l’analisi di fingerprinting molecolari ottenibili con il primer M13. L’approccio impiegato consente di ridurre considerevolmente il numero di individui da analizzare nel caso in cui si conducano indagini su comunità fungine complesse nelle quali sono comunemente presenti miceli sterili

Aiutare le vittime della mafia a ribellarsi

Riassunto. Nino Miceli è un imprenditore siciliano. Originario di Realmonte, in provincia di Agrigento, all’inizio degli anni ’90 è titolare della concessionaria Lancia di Gela. Si impegna molto nel suo lavoro e i risultati non tardano ad arrivare. La clientela aumenta, gli affari procedono, ma è proprio quando l’attività comincia a prosperare che la mafia gli presenta il conto. Deve pagare il “pizzo”, la tangente a cui ogni commerciante della zona deve sottostare. Nino Miceli non ci sta. Al principio subisce, ma presto inizia a registrare le conversazioni con gli esattori di Cosa Nostra per avere le prove del reato. Spinto e sostenuto da un ufficiale dei carabinieri, denuncia l’estorsione, subendo per questo minacce e ritorsioni. E’ costretto a trasferirsi, diviene un testimone di giustizia, cambia identità. All’inizio del 2007 esce il suo libro “Io, il fu Nino Miceli”. La rivista “Il Carabiniere” ha raccolto la sua testimonianza, pubblicata nel numero di aprile. Il paper analizzerà pertanto il ruolo dell’Arma di Carabinieri nel sostenere il percorso intrapreso dal soggetto che diviene, infine, in grado di ribellarsi e reagire ai ricatti ed alle estorsioni imposti dalla criminalità organizzata. Abstract Nino Miceli is a Sicilian man, owner of a small business: at the beginning of 1990s he is concessionaire of cars in Gela, an almost big town in Sicily. He works hard and soon he starts to gain a lot. Business is very good, customers are many and everything seems to proceed in the right direction. But the mafia keeps an eye on him and, at a certain point, it asks him to pay what in Italy is called “pizzo”. Nino Miceli does not want to pay it, he does not want to surrender to the mafia coercion. However, at the beginning he pays the “pizzo” but then he starts to record the conversation he has with those men (the “mafiosi”), in order to prove and demonstrate the crime. He also looks for protection at the local station of Carabinieri; finally, he decides to report the extortion but, from that time, he starts to experience more and more serious retorts and threats. Helped by an officier of Carabinieri, he becomes a witness of justice and, for that reason, he must change his identity. He leaves Sicily along with his family and move into a different region of Italy, far from his town, where nobody knows him. In 2007 he decides to write a book of memories, titled “Io, il fu Nino Miceli” (in English it could sound like “I was Nino Miceli”). This article tells his story, speaking also about the role played by the Arma of Carabinieri in supporting victims of racket and usury. Résumé Nino Miceli est un entrepreneur sicilien originaire de Realmonte, une ville en province d'Agrigento. Au début des années 90, il est proprietaire du garage Lancia à Gela. Il s'engage beaucoup dans son travail et les résultats ne tardent pas à arriver. La clientèle augmente, les affaires vont bon train, mais c'est justement quand l'entreprise est en train de prospérer que la mafia lui demande des comptes. Il doit payer le pot-de-vin comme tous les commerçants du quartier. Nino Miceli n'est pas d'accord avec cela. Au début il subit, mais il commence bientôt à enregistrer les conversations avec les percepteurs de Cosa Nostra pour collecter les preuves du crime. Il est poussé et soutenu par un officier des Carabinieri, il porte plainte pour extorsion, et à la suite de quoi il sera menacé et subira des rétorsions. Il doit déménager, il devient témoin de justice, il change d'identité. Au début de l'année 2007 son livre est publié avec le titre "Io, il fu Nino Miceli" (Moi, le fut Nino Miceli). La revue "Il Carabiniere" a recueilli son témoignage et l'a publié dans le numéro d'Avril 2007. Cette contribution analysera donc le rôle de l'Arme des Carabiniers dans le parcours entrepris par cet homme, qui devient enfin capable de se rebeller et de réagir aux chantages et aux extorsions imposés par la criminalité organisée

XMM-Newton evidence of shocked ISM in SN 1006: indications of hadronic acceleration

Shock fronts in young supernova remnants are the best candidates for being sites of cosmic ray acceleration up to a few PeV, though conclusive experimental evidence is still lacking. Hadron acceleration is expected to increase the shock compression ratio, providing higher postshock densities, but X-ray emission from shocked ambient medium has not firmly been detected yet in remnants where particle acceleration is at work. We exploited the deep observations of the XMM-Newton Large Program on SN 1006 to verify this prediction. We performed spatially resolved spectral analysis of a set of regions covering the southeastern rim of SN 1006. We studied the spatial distribution of the thermodynamic properties of the ambient medium and carefully verified the robustness of the result with respect to the analysis method. We detected the contribution of the shocked ambient medium. We also found that the postshock density of the interstellar medium significantly increases in regions where particle acceleration is efficient. Under the assumption of uniform preshock density, we found that the shock compression ratio reaches a value of ~6 in regions near the nonthermal limbs. Our results support the predictions of shock modification theory and indicate that effects of acceleration of cosmic ray hadrons on the postshock plasma can be observed in supernova remnants.Comment: Accepted for publication in A&

Suitability of borago officinalis for minimal processing as fresh-cut produce

Borage (Borago offcinalis L.) is a wild vegetable appreciated as a folk medicine and for culinary preparations. The introduction of borage as a specialized cultivation would allow for the diversification of vegetable crops and would widen the offerings of raw and minimally processed leafy vegetables. Thus, the aim of the research was to evaluate the quality and shelf-life of fresh-cut borage stored at different temperatures. Borage plants were grown during the autumn-winter season and immediately minimally processed after harvest. Fresh-cut borage leaves packed in sealed bags were stored at 2 or 6 °C for 21 d. Weight loss, total soluble solids (TSS), titratable acidity (TA), ascorbic acid, nitrates, leaf color characteristics and overall quality were determined through the storage period. Borage plants were deemed suitable for minimal processing. Storage temperature significantly influenced the rate of quality loss. Borage leaves had an initial nitrate content of 329.3 mg kg-1 FW that was not affected by temperature or storage. TSS and TA were higher in leaves stored at 6 °C. TSS, TA and ascorbic acid content increased during storage. Minimally processed borage leaves stored at 2 °C had lower weight loss and leaf color modifications during storage and a longer shelf life than those stored at 6 °C, so were still marketable after 21 d of storage

A Long Journey towards Citizenship: District Areas and Social Services in Goteborg

The main purpose of the present research is to investigate the implemented decentralized model of administration in local areas of Goteborg City and its effects on local welfare, or what I associate with the concept of social citizenship. In 1990 in Goteborg City the social sector - together with some other sectors (i.e. school and leisure) – was split up in 21 administrative District Councils. At that time politicians said that the main purpose of the district reform was to enhance social services management and citizenship. Citizenship entails a combination of participation and rights. This study will investigate the ways the reform improved local welfare. More precisely, it may be said that this is a study focusing on social administration mainly concerning the development of collective action and improved administrative performances for the advancement of local welfare. In Italy, instead of dealing with the increase of the complexity in social field through public intervention, the social services reform enacted by the law 328/2000 paved the way to partly privatize the Italian welfare state. This meant a much more massive presence of private actors in the social field. On the contrary, as it will be seen the Swedish government kept private subjects away from the management of social services by giving them the possibility to perform in political and cultural activities. The methodological design for this study can be complicated, since the implementation process could be seen as a journey that in Sweden started since 1952 and it is still in progress. Thus, I had to limit my purpose mainly in three questions: (1) How did the new institutional frame of reference contribute to improve social citizenship? (2) How did the district reform affect social services with regard to the building process of the local welfare? (3) In which way is Goteborg’s model to strengthen social citizenship distinguished from the Italian one? Our data consists of ten semi-structured interviews conducted thanks to the availability of key persons working in the municipality as city or district managers and social workers. The result is a clearer understanding of the district reform and its mechanism of functioning. For instance, it was possible to register the improvement of citizenship rights fulfilment due to the new local welfare model characterized by the institutional proximity to citizens, integration of different public services and sectors, coordination through decision-making decentralisation

The loss-limited electron energy in SN 1006: effects of the shock velocity and of the diffusion process

The spectral shape of the synchrotron X-ray emission from SN 1006 reveals the fundamental role played by radiative losses in shaping the high-energy tail of the electron spectrum. We analyze data from the XMM-Newton SN 1006 Large Program and confirm that in both nonthermal limbs the loss-limited model correctly describes the observed spectra. We study the physical origin of the observed variations of the synchrotron cutoff energy across the shell. We investigate the role played by the shock velocity and by the electron gyrofactor. We found that the cutoff energy of the syncrotron X-ray emission reaches its maximum value in regions where the shock has experienced its highest average speed. This result is consistent with the loss-limited framework. We also find that the electron acceleration in both nonthermal limbs of SN 1006 proceeds close to the Bohm diffusion limit, the gyrofactor being in the range 1.5-4. We finally investigate possible explanations for the low values of cutoff energy measured in thermal limbs.Comment: Accepted for publication in Astronomische Nachrichten. Proceedings of the XMM-Newton Science Workshop 201

Observability and diagnostics in the X-ray band of shock-cloud interactions in supernova remnants

X-ray emitting features originating from the interaction of supernova shock waves with small interstellar gas clouds are revealed in many X-ray observations of evolved supernova remnants (e.g. Cygnus Loop and Vela), but their interpretation is not straightforward. We develop a self-consistent method for the analysis and interpretation of shock-cloud interactions in middle-aged supernova remnants, which can provide the key parameters of the system and the role of relevant physical effects like the thermal conduction, without the need to run ad-hoc numerical simulations and to bother of morphology details. We explore all the possible values of the shock speed and cloud density contrast relevant to middle-aged SNRs with a set of hydrodynamic simulations of shock-cloud interaction, including the effects of thermal conduction and radiative cooling. From the simulations, we synthesize spatially and spectrally resolved focal-plane data as they would be collected with XMM-Newton/EPIC, an X-ray instrument commonly used in these studies. We devise and tune up two diagnostic tools, the first based on the mean-photon energy vs. count rate scatter plot and the second on the spectral analysis of the interaction region, that can be used to highlight the effects of thermal conduction and to derive the shock speed in case of efficient conduction at work. These tools can be used to ascertain information from X-ray observations, without the need to develop detailed and ad-hoc numerical models for the interpretation of the data.Comment: 9 pages, 7 Figures; accepted for publication on A&A. Version with full resolution images can be found at http://www.astropa.unipa.it/~orlando/PREPRINTS/sorlando_13801.pd

New Insights into SNR Evolution Revealed by the Discovery of Recombining Plasmas

We report the discovery of recombining plasmas in three supernova remnants (SNRs) with the Suzaku X-ray astronomy satellite. During SNR's evolution, the expanding supernova ejecta and the ambient matter are compressed and heated by the reverse and forward shocks to form an X-ray emitting hot plasma. Since ionization proceeds slowly compared to shock heating, most young or middle-aged SNRs have ionizing (underionized) plasmas. Owing to high sensitivity of Suzaku, however, we have detected radiative recombination continua (RRCs) from the SNRs IC 443, W49B, and G359.1-0.5. The presence of the strong RRC is the definitive evidence that the plasma is recombining (overionized). As a possible origin of the overionization, an interaction between the ejecta and dense circumstellar matter is proposed; the highly ionized gas was made at the initial phase of the SNR evolution in dense regions, and subsequent rapid adiabatic expansion caused sudden cooling of the electrons. The analysis on the full X-ray band spectrum of IC 443, which is newly presented in this paper, provides a consistent picture with this scenario. We also comment on the implications from the fact that all the SNRs having recombining plasmas are correlated with the mixed-morphology class.Comment: Published by Advances in Space Researc

Spatial identification of the overionized plasma in W49B

Recent Suzaku X-ray observations of the ejecta-dominated supernova remnant W49B have shown that in the global spectrum there is a clear indication for the presence of overionized plasma whose physical origin is still under debate. In order to ascertain the physical origin of such a rapidly cooling plasma, we focus on the study of its spatial localization within the X-ray emitting ejecta. We confirm the presence of a saw-edged excess (interpreted as a strong radiative recombination continuum) in the global spectrum above 8 keV, emerging above the ionization-equilibrium model. We produce a hardness ratio map to determine where the plasma is overionized and we perform a spectral analysis of the regions with and without strong overionization. We find that the overionized plasma is localized in the center of the remnant and in its western jet, while it is not detected in the bright eastern jet, where the expansion of the ejecta is hampered by their interaction with a dense interstellar cloud. The location of overionized plasma suggests that the inner ejecta are rapidly cooling by expansion, unlike the outer ejecta, for which expansion is hampered by interstellar clouds seen in H2Comment: Accepted for publication as a Letter in Astronomy and Astrophysics