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Extracorporeal blood purification therapies are increasingly applied in the field of intensive care medicine. These techniques are applied to a relevant number of clinical situations as Sepsis, Trauma, burns, Influenza, surgery, Pancreatitis, ARDS, Life support, Liver failure. Compared to filtration-based methods mainly used for renal replacement therapy, newest adsorptive approaches have shown to specifically target the inflammatory cascade by the effective removal of relevant mediators. In the neonatal and pediatric setting however, the application of these methods brings with it various challenges but also profound technical difficulties. Recently, a promising extracorporeal device for cytokine adsorption (Cytosorb) has been introduced, however data for its application in critically ill pediatric patients remains sparse [1]. The lack of data and procedure protocols led us to take some aprioristic diagnostic and speculative decisions only from a scientific point of view: “in primis” when we decided for the first time in a pediatric patient to use this "Cytosorb Absorber", a device not registered and delivered for toddlers; second when it was clear that we could extend its use to not conventional disease therapeutic strategies; third when we decided to give to “Cytosorb” use the appellation and indication of “rescue treatment”. These results are interesting for the scientific community, but further research is needed.

Natural mortality of the almond bark beetle, Scolytus amygdali Guerin-Meneville (Coleoptera: Curculionidae: Scolytinae) was studied during its spring generation under the bark of infested almond tree branches in Tunisia. Dead larvae and adults of the beetle were collected from the field and analyzed under the laboratory conditions. Two main fungal isolates were obtained from the cadavers of S. amygdali individuals,identified using morphological and molecular tools, as Aspergillus flavus and Fusarium oxysporum. In preliminary studies conducted under laboratory conditions, species of Aspergillus and Fusarium fungi were reported virulent at the concentrations of (0.4 x 107 conidia/ml) to S. amygdali. In order to test the pathogencity of the two isolated fungal species, similar concentrations were applied on larvae and adults of healthy S. amygdale individuals. Both fungi were found significantly virulent immediately after 24h under laboratory conditions. Further investigations for their suitability as biocontrol agents in the field are required.

Extracorporeal blood purification therapies are increasingly applied in the field of intensive care medicine. These techniques are applied to a relevant number of clinical situations as Sepsis, Trauma, burns, Influenza, surgery, Pancreatitis, ARDS, Life support, Liver failure. Compared to filtration-based methods mainly used for renal replacement therapy, newest adsorptive approaches have shown to specifically target the inflammatory cascade by the effective removal of relevant mediators. In the neonatal and pediatric setting however, the application of these methods brings with it various challenges but also profound technical difficulties. Recently, a promising extracorporeal device for cytokine adsorption (Cytosorb) has been introduced, however data for its application in critically ill pediatric patients remains sparse1. The lack of data and procedure protocols led us to take some aprioristic diagnostic and speculative decisions only from a scientific point of view: “in primis” when we decided for the first time in a pediatric patient to use this "Cytosorb Absorber", a device not registered and delivered for toddlers; second when it was clear that we could extend its use to not conventional disease therapeutic strategies; third when we decided to give to “Cytosorb” use the appellation and indication of “rescue treatment”. These results are interesting for the scientific community, but further research is needed.

Phenolic compounds are plant secondary metabolites that are important determinants in both sensory and nutritional quality of fruits and vegetables. In addition, in the last few years many studies studies support the increasing evidence of the possible role of phenolic compounds in prevention of chronic diseases such as cardiovascular disease and cancer [Nakajima et al., 2007, Life Sciences 80: 370–377]. Tomatoes are one of the valuable sources of antioxidants such as phenolic compounds (flavonoids and hydroxycinnamic acid derivatives). Therefore, increasing phenolics in tomato fruit is a major claim of breeding in order to meet consumer requirements and create new market opportunities. So far, many key genes have been reported that control the accumulation in planta of phenolic compounds. However, additional insights are required in order to highlight genetic and physiological mechanisms controlling phenols accumulation in tomato fruit and to support breeding programs for increased fruit quality. The aim of the work was the identification of major genes and gene networks that regulate the level of phenols in tomato fruit. The screening of Solanum pennellii x S. lycopersicum introgression lines (ILs) [Eshed and Zamir, 1995, Genetics 141:1147-1162] over three year trials in greenhouse and open field environments allowed the identification of a stable QTL for increased fruit content of total phenolics in the IL7-3. In particular, chlorogenic acid mainly accounted for the higher performance of this line. Therefore, to investigate candidate genes controlling phenols synthesis and accumulation in IL7-3 fruit, we performed a comparative transcriptomic analysis in tomato pericarp between this line and the control cv. M82 over two consecutive years. The transcriptomic approach allowed to identify 149 up-rgulated and 142 down-regulated probes. Based on functional annotation, clustering and networking outputs, subsets of differentially expressed transcripts were used to develop model networks that describe mechanisms controlling accumulation of phenylpropanoids in tomato fruit. The network explains the variation in phenols levels in terms of interactions between ethylene signalling, plant responses to stress and biosynthesis of phenolics. Upon validation of key transcripts of our model by RT-qPCR we undertook a functional characterization of candidate genes by the TILLING (Targeting Induced Local Lesion IN Genomes) approach [Minoia et al., 2010, BMC Research Notes 3:69]. In particular, tomato plants homozygous for mutations in the coding sequence of two transcription factors involved in ethylene response (e.g. ERF1 and EIN3) revealed to drive ripening-associated phenol accumulation in fruit. These results suggested us to design new strategies of precision breeding for increased fruit nutritional quality in tomato.

L’obiettivo del progetto ha riguardato la combinazione di QTL coinvolti nell’incremento del contenuto in antiossidanti nel frutto di pomodoro. A tale scopo sono state utilizzate le linee di introgressione (IL) di pomodoro. Tali IL sono particolarmente utili per la dissezione di caratteri complessi e consentono l’individuazione ed il mappaggio di QTL di interesse su specifici cromosomi. Il trasferimento dei QTL e la loro combinazione in nuovi genotipi è stato perseguito eseguendo incroci tra linee di introgressione. L’ibrido F1 recante le due introgressioni allo stato eterozigote è stato sottoposto ad autofecondazione ottenendo una progenie F2 nell’ambito della quale si è provveduto alla identificazione delle piante recanti le introgressioni desiderate allo stato omozigote mediante un approccio di selezione assistita da marcatori molecolari (MAS). In particolare, la IL7-3 e la IL12-4 sono state selezionate in quanto riportate esprimere rispettivamente un maggiore contenuto in fenoli totali (Phe) ed ascorbato (AsA) nel pericarpo mentre IL2-2 e IL6-3 sono state selezionate in quanto riportate esprimere un maggior contenuto in carotenoidi totali (Car) nel frutto. Dallo screening molecolare della popolazione derivante dall’incrocio IL12-4xIL7-3 si sono ottenute quattro piante doppio omozigoti. Le piante sono state allevate in due condizioni di crescita diverse (serra e pieno campo) e sono state sottoposte all’analisi fenotipiche per valutare il contenuto di AsA, fenoli totali e solidi solubili. I doppi omozigoti hanno mostrato un elevato contenuto di AsA, fenoli e solidi solubili nel frutto, rispetto al genotipo controllo, dimostrando la validità dell'approccio del pyramiding per incrementare il contenuto di antiossidanti nel frutto. Mediante l'analisi molecolare sono state selezionate anche 4 sub-linee della regione 12-4, che potranno essere utili in futuro per identificare geni candidati al controllo dell'AsA. Analogamente, le piante doppio omozigoti isolate dalla progenie dell’incrocio IL6-3xIL2-2 ed i rispettivi parentali sono state allevate in serra e sono state effettuate analisi chimico-nutrizionali. Tali campioni, infatti, sono stati sottoposti all’analisi HPLC per verificare il contenuto in carotenoidi nei frutti. Dall’analisi eseguita si è evidenziato un incremento del livello di β-carotene nelle linee doppio omozigoti e contemporaneamente una diminuzione del contenuto in trans-licopene. Tutti gli altri componenti non hanno evidenziato differenze statisticamente significative. Inoltre, campioni di frutto sono stati raccolti dalla sub-linea IL2-2-1 e da M82 a cinque diversi stadi di maturazione e sono stati sottoposti ad analisi metabolica e trascrittomica per il contenuto in carotenoidi. I risultati hanno evidenziato un incremento del contenuto di β carotene nei frutti della IL2-2-1 durante tutto il ciclo di maturazione ed una riduzione del contenuto di cis-licopene. Allo scopo di identificare variazioni trascrizionali di geni coinvolti nella sintesi dei carotenoidi è stata condotta una quantificazione relativa mediante Real Time qPCR di 12 geni chiave della biosintesi dei carotenoidi. L’analisi statistica dei dati ottenuti ha evidenziato differenze esclusivamente nell’abbondanza relativa del trascritto del gene β-carotene idrossilasi rispetto ad M82.