Studio fitochimico di Clerodendrum splendens G. Don (Verbenaceae)

Oggetto della presente tesi è lo studio fitochimico delle parti aeree di Clerodendrum splendens G. Don, un arbusto rampicante sempreverde appartenente alla famiglia delle Verbenaceae, originaria dell’ Africa occidentale ed acclimatato presso El Zoharia Research Garden del Cairo in Egitto. Lo studio di piante provenienti da tale giardino botanico rientra in un progetto di ricerca in collaborazione con la Faculty of Pharmacy, Al Azhar University del Cairo, Egitto, progetto che prevede l’isolamento e la caratterizzazione chimica di metaboliti secondari presenti in diverse specie vegetali mediante tecniche cromatografiche e spettroscopiche. L’ interesse verso questa specie nasce in primo luogo dall’esiguo numero di studi svolti su di essa ed inoltre, sulla base di dati presenti in letteratura sul genere, allo scopo di isolare e caratterizzare nuovi composti di natura diterpenica quali possibili prodotti di interesse biologico. Il materiale vegetale (parti aeree) è stato sottoposto a essiccazione all’aria, macinazione e successiva estrazione a temperatura ambiente con solventi a polarità crescente: n-esano, cloroformio, cloroformio–metanolo (9:1) e metanolo. In questo studio sono stati presi in considerazione gli estratti: cloroformico (RCHCl3), cloroformio–metanolo (RC-M), e metanolico (RMeOH). Il residuo RC-M è stato è stato sottoposto a cromatografia su colonna Sephadex LH-20 ad esclusione molecolare e, successivamente, le frazioni di interesse sono state studiate tramite cromatografia ad alta pressione su fase inversa (RP-HPLC), giungendo alla purificazione di 2 derivati fenolici, il composto flavonoidico ispidulina 7-O- β-D-glucopiranoside (1), ed il fenilpropanoide phlinoside B (2). Il residuo RMeOH è stato ripartito in una frazione acquosa ( RW ) ed in una n-butaolica ( RBu ) che è stata sottoposta a cromatografia su colonna Sephadex LH-20 ad esclusione molecolare. In seguito le frazioni di interesse sono state studiate tramite RP-HPLC giungendo all’ isolamento di 8 metaboliti secondari tra cui un nuovo fenilpropanoide (3), (3,4-diidrossifenil)etil-O-β-D-xilopiranosil-(1→2)-α-L-ramnopiranosil-(1→3)-6-O-t-caffeoil-β-D-glucopiranoside , i composti (1) e (2) già isolati nel residuo RCM , ed altri composti fenolici tra cui ispidulina 7-neoesperidoside (4), verbascoside (5), isoacteoside (6), luteolina 7-neoesperidoside (7), ed infine acido rosmarinico (8) . Il residuo RCHCl3 è stato sottoposto a cromatografia flash su colonna di gel di silice e, in seguito, le frazioni di interesse sono state studiate tramite cromatografia ad alta pressione su fase inversa RP-HPLC giungendo alla purificazione di composti di origine terpenica, fra cui un composto sesquiterpenico 2β-angeloilossi-5β-idrossi-7αH,10β-metil-eudesm-3-en-1-one (9), il diterpene 14,15-diidro-15-idrossi-3-epicarioptina (10), e 4 composti diterpenici mai precedentemente isolati, il 2-acetossi-3-(2’,3’-diacetossi-2’-metil)-butanoilossi-14-idro-15-idrossiclerodina (11), il 3,15-diidrossi-14-idro-clerodina (12), il 2,15-diidrossi-3-(2’-idrossi-2’-metil-3’-acetossi)-butanoilossi-6α,18-diacetossi-4α,17-epossi-clerodan-11,16-lattone (13), ed infine il diterpene 3,14,15-tridrossi-6α,18-diacetossi-4α,17-epossi-clerodan-11,16-lattone (14) . La determinazione strutturale di tutti i composti è stata effettuata con l’ausilio di tecniche spettroscopiche NMR mono e bidimensionali, fra cui 1H-NMR, 13C-NMR, HSQC, HMBC, 1D-TOCSY sono state le principali, e tecniche di spettrometria di massa ESI-MS. L’indagine fitochimica condotta su Clerodendrum splendens G. Don ha quindi portato all’isolamento totale di 14 composti tra cui 4 diterpeni mai caratterizzati in precedenza, 1 nuovo fenilpropanoide ed altri derivati fenolici nel completo accordo con quanto trovato negli studi riportati nella letteratura appartenente al genere Clerodendrum

JRC technical work supporting Commission second level legislation on risk based contributions to the (single) resolution fund

JRC supported the DG MARKT by developing quantitative analyses for the preparation of the second level legislation on bank contributions to be paid to the EU national Resolution Funds and to the Single Resolution Fund SRF for countries participating to the Banking Union. The present report summarizes all the extensive analyses on the calculation of banks contributions supporting the whole policy process. All analyses were based on a dataset that JRC built assembling individual bank unconsolidated balance sheet data, provided directly by the MS. JRC developed the technical details to measure the risk profile of each bank. Starting from a selection of balance sheet indicators, which account for the different aspects of each bank activity, the methodology aggregates them into a single composite risk indicator. The risk indicator is then combined with the bank size measure to compute the share of aggregated contribution each bank joining the fund would pay. JRC also investigated the decrease in contributions of applying a special treatment for the computation of the small banks’ contributions: these banks will not pay contributions based on their risk profile but will be instead lump-sum contributions, depending on their size only. JRC assessed the sensitivity of the distribution of contributions when changing some elements of the overall mechanism used to measure risk and compute contributions. Finally, following the discussion at the political level, JRC also assessed some technical issues related to the calculation of the contribution base and it tested the impact on banks contributions of different options for the phasing in of the single resolution fund.JRC.G.1-Financial and Economic Analysi

Interaction between <i>Mycobacterium tuberculosis</i>, <i>Mycobacterium bovis</i>, <i>Mycobacterium avium</i> subspecies <i>paratuberculosis</i> with the enteric glia and microglial cells

Background We investigated the interaction of Mycobacterium avium subspecies paratuberculosis, M. bovis and M. tuberculosis and different glial cells (enteric glial and microglial cells) in order to evaluate the infecting ability of these microorganisms and the effects produced on these cells, such as the evaluation of cytokines expression. Results Our experiments demonstrated the adhesion of M. paratuberculosis to the enteroglial cells and the induction of IL-1A and IL-6 expression; M. tuberculosis and M. bovis showed a good adhesive capability to the enteric cell line with the expression of the following cytokines: IL-1A and IL-1B, TNF-α, G-CSF and GM-CSF; M. bovis induced the expression of IL-6 too. The experiment performed with the microglial cells confirmed the results obtained with the enteroglial cells after the infection with M. tuberculosis and M. bovis, whereas M. paratuberculosis stimulated the production of IL-1A and IL-1B. Conclusion Enteroglial and microglial cells, could be the target of pathogenic mycobacteria and, even if present in different locations (Enteric Nervous System and Central Nervous System), show to have similar mechanism of immunomodulation

Deep Image Prior Amplitude SAR Image Anonymization

This paper presents an extensive evaluation of the Deep Image Prior (DIP) technique for image inpainting on Synthetic Aperture Radar (SAR) images. SAR images are gaining popularity in various applications, but there may be a need to conceal certain regions of them. Image inpainting provides a solution for this. However, not all inpainting techniques are designed to work on SAR images. Some are intended for use on photographs, while others have to be specifically trained on top of a huge set of images. In this work, we evaluate the performance of the DIP technique that is capable of addressing these challenges: it can adapt to the image under analysis including SAR imagery; it does not require any training. Our results demonstrate that the DIP method achieves great performance in terms of objective and semantic metrics. This indicates that the DIP method is a promising approach for inpainting SAR images, and can provide high-quality results that meet the requirements of various applications

Tuberculosis in Sardinia: An investigation into the relationship between natives and immigrants

AbstractObjective/background: Tuberculosis (TB) has had a recrudescence in the last few decades in Italy as a result of many factors, among which migration from countries where TB is endemic is one of them. In Sardinia, a major island of Italy, there was no knowledge of the mechanisms of transmission of TB in the immigrant subpopulation and the impact it may have on the native subpopulation and on the community as a whole. Therefore, a molecular epidemiological study was carried out to get a clearer picture of the number and genetic features of Mycobacterium tuberculosis strains isolated from immigrants and from natives in Sardinia. Methods: Two groups of clinical isolates of M. tuberculosis, one collected from immigrants and the other one from Sardinians, were analyzed in this study. The genotyping was executed through the variable number tandem repeat-mycobacterial interspersed repetitive units technique and a first-line antimycobacterial drug-susceptibility test was also carried out. Results: Thirty-six clinical isolates from immigrants and 25 from Sardinians were analyzed. Variable number tandem repeat-mycobacterial interspersed repetitive units technique showed that all of them belonged to different strains and there was a quite high allelic diversity among them. Moreover, data collected allowed the finding of, with a good approximation, the phylogenetic relations among the strains isolated and the best-known phylogenetic groups. Conclusion: The study pointed out that since every strain is different, there was no TB transmission in any of the subpopulations and between immigrants and natives. This showed that the presence of immigrants was not a risk factor for contracting TB in the community

"In vitro" activities of antimycobacterial agents against <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> linked to Crohn's disease and paratuberculosis

Crohn's disease, a human disease similar to paratuberculosis in animals is the most painful and devastating disease that may involve infection with M. avium subsp. paratuberculosis (MAP), different genetic polymorphisms and an immune dysregulation syndrome. Treatment of Crohn's disease is most commonly based on 5-aminosalicylic acid (5-ASA) compounds, corticosteroids, and immunosuppressive agents. Recently, biological therapies using monoclonal antibodies against inflammatory cytokines have shown some positive results. However, all these therapies treat the symptoms not the cause of the disease

Evaluation of the Antimicrobial Properties of the Essential Oil of Myrtus communis L. against Clinical Strains of Mycobacterium spp.

Mycobacterium tuberculosis is the etiological agent of tuberculosis. The World Health Organization has estimated that 8 million of people develop active TB every year and the situation is complicated by an increase of Mycobacterium tuberculosis strains resistant to drugs used in antitubercular therapy: MDR and XDR-TB. Myrtle leaf extracts, used as an antiseptic in Sardinian traditional medicine, have strong antibacterial activity as several investigations showed. In this study we investigated the antimicrobial properties of the essential oil of Myrtus communis against clinical strains of M. tuberculosis and M. paratuberculosis

Interaction between Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium avium subspecies paratuberculosis with the enteric glia and microglial cells

<p>Abstract</p> <p>Background</p> <p>We investigated the interaction of <it>Mycobacterium avium </it>subspecies <it>paratuberculosis, M. bovis </it>and <it>M. tuberculosis </it>and different glial cells (enteric glial and microglial cells) in order to evaluate the infecting ability of these microorganisms and the effects produced on these cells, such as the evaluation of cytokines expression.</p> <p>Results</p> <p>Our experiments demonstrated the adhesion of <it>M. paratuberculosis </it>to the enteroglial cells and the induction of IL-1A and IL-6 expression; <it>M. tuberculosis </it>and <it>M. bovis </it>showed a good adhesive capability to the enteric cell line with the expression of the following cytokines: IL-1A and IL-1B, TNF-α, G-CSF and GM-CSF; <it>M. bovis </it>induced the expression of IL-6 too.</p> <p>The experiment performed with the microglial cells confirmed the results obtained with the enteroglial cells after the infection with <it>M. tuberculosis </it>and <it>M. bovis</it>, whereas <it>M. paratuberculosis </it>stimulated the production of IL-1A and IL-1B.</p> <p>Conclusion</p> <p>Enteroglial and microglial cells, could be the target of pathogenic mycobacteria and, even if present in different locations (Enteric Nervous System and Central Nervous System), show to have similar mechanism of immunomodulation.</p