The glucocorticoid PYED-1 disrupts mature biofilms of Candida spp. and inhibits hyphal development in Candida albicans

Invasive Candida infections have become a global public health problem due to the increase of Candida species resistant against antifungal therapeutics. The glucocorticoid PYED-1 (pregnadiene-11-hydroxy-16α,17α-epoxy-3,20-dione-1) has antimicrobial activity against various bacterial taxa. Consequently, it might be considered for the treatment of Candida infections. The antifungal activity of PYED-1 was evaluated against several fungal strains that were representative of the five species that causes the majority of Candida infections—namely, Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis and Candida krusei. PYED-1 exhibited a weak antifungal activity and a fungistatic effect on all five Candida species. On the other hand, PYED-1 exhibited a good antibiofilm activity, and was able to eradicate the preformed biofilms of all Candida species analyzed. Moreover, PYED-1 inhibited germ tube and hyphae formation of C. albicans and reduced adhesion of C. albicans to abiotic surfaces by up to 30%

Physical activity regulates tnfα and il-6 expression to counteract inflammation in cystic fibrosis patients

Background: Cystic fibrosis (CF) is one of the most common inherited diseases. It is characterised by a severe decline in pulmonary function associated with metabolic perturbations and an increased production of inflammatory cytokines. The key role of physical activity (PA) in improving the health status of CF patients and reducing lung function decline has recently been demonstrated. This study evaluated interleukin-6 (IL-6) and tumour necrosis factor α (TNFα) expression in two subgroups of CF patients classified based on PA. Methods: We selected 85 CF patients; half of them regularly undertook supervised PA in the three years leading up to the study and half of them were not physically active. Patients were analysed for serum IL-6 and TNFα levels using enzyme-linked immunosorbent assays. Results: We found that the expression levels of IL-6 and TNFα differed in terms of their regulation by PA. In particular, TNFα levels negatively correlated with FEV1% decrease/year and FEV1% decrease (p = 0.023 and p = 0.02, respectively), and positively correlated with serum fasting glucose (p = 0.019) in PA CF patients. In contrast, in the NPA subgroup, TNFα levels were positively correlated with IL-6 (p = 0.001) and negatively correlated with adiponectin (p = 0.000). In addition, multiple logistic regression analysis confirmed that PA is an independent modulator of the inflammatory state. Conclusions: PA modulates inflammatory processes in CF patients by regulating the secretion of pro-inflammatory cytokines and thus ameliorating lung function. Our data show that PA is a useful complementary strategy in the management of CF and that TNFα may be a marker of these effects of PA

N-nonyloxypentyl-l-deoxynojirimycin inhibits growth, biofilm formation and virulence factors expression of Staphylococcus aureus

Staphylococcus aureus is one of the major causes of hospital-and community-associated bacterial infections throughout the world, which are difficult to treat due to the rising number of drug-resistant strains. New molecules displaying potent activity against this bacterium are urgently needed. In this study, d-and l-deoxynojirimycin (DNJ) and a small library of their N-alkyl derivatives were screened against S. aureus ATCC 29213, with the aim to identify novel candidates with inhibitory potential. Among them, N-nonyloxypentyl-l-DNJ (l-NPDNJ) proved to be the most active compound against S. aureus ATCC 29213 and its clinical isolates, with the minimum inhibitory concentration (MIC) value of 128 µg/mL. l-NPDNJ also displayed an additive effect with gentamicin and oxacillin against the gentamicin-and methicillin-resistant S. aureus isolate 00717. Sub-MIC values of l-NPDNJ affected S. aureus biofilm development in a dose-dependent manner, inducing a strong reduction in biofilm biomass. Moreover, real-time reverse transcriptase PCR analysis revealed that l-NPDNJ effectively inhibited at sub-MIC values the transcription of the spa, hla, hlb and sea virulence genes, as well as the agrA and saeR response regulator genes

Trans-heterozygosity for mutations enhances the risk of recurrent/chronic pancreatitis in patients with Cystic Fibrosis

Background: Recurrent (RP) and chronic pancreatitis (CP) may complicate Cystic Fibrosis (CF). It is still unknown if mutations in genes involved in the intrapancreatic activation of trypsin (IPAT) or in the pancreatic secretion pathway (PSP) may enhance the risk for RP/CP in patients with CF.Methods: We enrolled: 48 patients affected by CF complicated by RP/CP and, as controls 35 patients with CF without pancreatitis and 80 unrelated healthy subjects. We tested a panel of 8 genes involved in the IPAT, i.e. PRSS1, PRSS2, SPINK1, CTRC, CASR, CFTR, CTSB and KRT8 and 23 additional genes implicated in the PSP.Results: We found 14/48 patients (29.2%) with mutations in genes involved in IPAT in the group of CF patients with RP/CP, while mutations in such genes were found in 2/35 (5.7%) patients with CF without pancreatitis and in 3/80 (3.8%) healthy subjects (p < 0.001). Thus, we found mutations in 12 genes of the PSP in 11/48 (22.9%) patients with CF and RP/CP. Overall, 19/48 (39.6%) patients with CF and RP/CP showed one or more mutations in the genes involved in the IPAT and in the PSP while such figure was 4/35 (11.4%) for patients with CF without pancreatitis and 11/80 (13.7%) for healthy controls (p < 0.001).Conclusions: The trans-heterozygous association between CFTR mutations in genes involved in the pathways of pancreatic enzyme activation and the pancreatic secretion may be risk factors for the development of recurrent or chronic pancreatitis in patients with CF

Incorporating weekly carboplatin in anthracycline and paclitaxel-containing neoadjuvant chemotherapy for triple-negative breast cancer: propensity-score matching analysis and TIL evaluation

Background The generation of data capturing the risk-benefit ratio of incorporating carboplatin (Cb) to neoadjuvant chemotherapy (NACT) for triple-negative breast cancer (TNBC) in a clinical practice setting is urgently needed. Tumour-infiltrating lymphocytes (TILs) have an established role in TNBC receiving NACT, however, the role of TIL dynamics under NACT exposure in patients receiving the current standard of care is largely uncharted. Methods Consecutive TNBC patients receiving anthracycline-taxane [A-T] +/- Cb NACT at three Institutions were enrolled. Stromal-TILs were evaluated on pre-NACT and residual disease (RD) specimens. In the clinical cohort, propensity-score-matching was used to control selection bias. Results In total, 247 patients were included (A-T = 40.5%, A-TCb = 59.5%). After propensity-score-matching, pCR was significantly higher for A-TCb vs A-T (51.9% vs 34.2%, multivariate: OR = 2.40, P = 0.01). No differences in grade &gt;= 3 haematological toxicities were observed. TILs increased from baseline to RD in the overall population and across A-T/A-TCb subgroups. TIL increase from baseline to RD was positively and independently associated with distant disease-free survival (multivariate: HR = 0.43, P = 0.05). Conclusions We confirmed in a clinical practice setting of TNBC patients receiving A-T NACT that the incorporation of weekly Cb significantly improved pCR. In addition, A-T +/- Cb enhanced immune infiltration from baseline to RD. Finally, we reported a positive independent prognostic role of TIL increase after NACT exposure

Le reti sismica e geodetica di pronto intervento dell’INGV: un primo impiego a seguito del terremoto de L’Aquila del 6 aprile 2009

Durante gli ultimi due anni l’Istituto Nazionale di Geofisica e Vulcanologia (INGV) ha sviluppato un importante infrastruttura di pronto intervento (la Rete Mobile Real-Time di Pronto Intervento), al fine di incrementare il numero di stazioni della Rete Sismica Nazionale dell’INGV (RSN) in zona epicentrale a seguito di eventi sismici rilevanti. Gli obiettivi principali della Rete Mobile Real-Time di Pronto Intervento sono il miglioramento delle localizzazioni epicentrali calcolate dalla Sala di Monitoraggio dell’INGV e l’abbassamento della soglia di detezione della micro-sismicità in area epicentrale durante una sequenza sismica. La Rete Mobile Real-Time di Pronto Intervento è composta da stazioni sismiche remote i cui dati sono telemetrati tramite ponte radio UHF (Ultra High Frequency) presso dei centri d’acquisizione intermedi (definiti “sottonodi”). I sottonodi sono a loro volta connessi tramite Wi-Fi ad un “centro stella” (nodo), ove è situato un sistema di trasmissione satellitare (Libra VSAT Nanometrics), tramite il quale vengono inviati i dati in tempo reale al centro acquisizione della Sala di Monitoraggio dell’INGV di Roma. L’acquisizione dati è ridondata inoltre presso la sala Disaster Recovery dell’Osservatorio di Grottaminarda. Il sistema d’acquisizione di dati sismici è costituito da un datalogger a tre canali, equipaggiato con un convertitore AD ad alta risoluzione (a 24 bit), dotato di un clock di precisione basato su timing GPS. I sensori sismici utilizzati presso le stazioni remote sono accelerometri Episensor FBA ES-T (Kinemetrics) con fondo scala a 2G e velocimetri a corto periodo (Lennartz Le Lite 3D). Il sistema di trasmissione dati, come accennato, si avvale di diversi apparati installati presso le stazioni remote, i sottonodi, ed il centro stella. Presso le stazioni remote è installato un radio modem operante in banda UHF (da 380 a 470 MHz), per il trasferimento trasparente di dati asincroni in modalità half-duplex. L’apparato modula in etere a 9.600 bps, realizzando collegamenti da 2 a 50 chilometri, in funzione dell’orografia locale e del sistema d’antenna utilizzato. Presso i sottonodi viene utilizzato un apparato WiFi (Wireless Fidelity) operante con frequenza di 2.4 GHz per collegamenti IP fino a 54 Mbit/s. Presso i sottonodi i dati sismici ricevuti dalle stazioni remote vengono inviati, tramite ponte Wi-Fi, al centro stella. Presso il centro stella la trasmissione dati avviene tramite il ricetrasmettitore Cygnus Nanometrics. Esso permette l’invio dei dati ricevuti alla Sala di Monitoraggio tramite collegamento satellitare. Il protocollo di trasmissione satellitare dedicato sul link VSAT è di tipo IP, ma può avvenire anche su apparati esterni quali fibra ottica, linee telefoniche, ecc. Per conseguire una maggiore flessibilità d’impiego, tale sistema dispone di due differenti frequenze di trasmissione, disponibili su satellite Intelsat ed HellaSat. Tutto ciò permette di orientare la parabola in due diverse direzioni, in modo da poter ovviare l’eventuale presenza di ostacoli come alberi, montagne o edifici. L’intera struttura racchiude queste tre diverse tecnologie di trasmissione dati (UHF, Wi-Fi e satellitare) al fine di garantire maggiore flessibilità di utilizzo; questo permette di affrontare l’emergenza sismica in tutte le condizioni logistiche e/o meteorologiche mirando a rapidi tempi di intervento (raggiungimento della zona epicentrale e istallazione). L’installazione della Rete Mobile Real-Time di Pronto Intervento viene gestita e coordinata all’interno di un Sistema Informativo Geografico (GIS) che consente la scelta della disposizione geografica ottimale delle stazioni della rete di pronto intervento intorno all’area epicentrale. Il database geografico utilizzato durante l’emergenza sismica contiene informazioni territoriali di vario tipo in area epicentrale. L’INGV dispone infatti di database geografici contenenti dati territoriali di tutto il territorio nazionale le cui categorie, utili ai fini della gestione dell’emergenza sismica, sono: Ubicazione delle stazioni delle reti di monitoraggio; Cartografia topografica IGM (1:25000, 1:50000, 1:100000); Modello digitale del terreno IGM; Uso del suolo; Viabilità e grafo stradale; Catologhi di sismicità storica e strumentale; Mappe di pericolosità sismica e del territorio; Database delle Sorgenti sismogenetiche; Mappe di scuotimento; Mappe di osservazioni macrosismiche. I dati sopra elencati sono utilizzati per la realizzazione di analisi di superficie (surface spatial analysis, Viewshed, Observer Point) che consentono la produzione di scenari utili per l’individuazione delle aree più favorevoli alla collocazione degli apparati della rete Real Time. Il terremoto de L’Aquila del 6 aprile 2009 è stato il primo caso di utilizzo dell’intera infrastruttura di pronto intervento. A meno di 6 ore dalla scossa principale (Mw 6.3 delle ore 01:32 GMT) il primo accelerometro inviava già dati alla Sala di Monitoraggio dell’INGV di Roma. A 3 giorni dall’evento la struttura di pronto intervento installata era costituita da 9 stazioni sismiche real-time. Oltre alla Rete Real Time di Pronto Intervento l’INGV ha installato 5 nuove stazioni GPS permanenti nel territorio abruzzese a seguito dell’evento del 6 aprile (Fig. 3). Le stazioni GPS permanenti presenti nel settore aquilano precedentemente al terremoto erano infatti caratterizzate da un’interdistanza troppo elevata, tale da non consentire una risoluzione spaziale adeguata del campo di spostamento co- e postsismico. A poche ore di distanza dall’evento sismico del 6 aprile si è quindi attivata una squadra di pronto intervento dell’INGV coadiuvata anche da personale del DPC-Ufficio Sismico e dell’ISPRA. A partire dal 7 aprile 2009, e fino al 17 dello stesso mese, sono state installate 5 nuove stazioni GPS permanenti (3 stazioni appartenenti alla Rete Integrata Nazionale GPS dell’INGV, 1 stazione del DPC-Ufficio Sismico ed una stazione dell’ISPRA) nei settori limitrofi all’epicentro della scossa principale della sequenza dell’aquilano. In tutte e 5 i casi la stazione GPS è stata monumentata, installata e avviata nell’arco di 5-6 ore. Su tutte le stazioni GPS è stata impostata sia un’acquisizione del dato GPS a 30 secondi sia un ringbuffer con campionamento a 10 Hz, in modo da permettere la registrazione dell’intera deformazione cosismica (sia statica che dinamica) in caso di ulteriore evento sismico. Nelle settimane successive è stata poi ottimizzata la trasmissione dei dati GPS, utilizzando un sistema di trasmissione dati via GPRS/UMTS implementato dal ST-Osservatorio di Grottaminarda.PublishedTrieste- Italy1.1. TTC - Monitoraggio sismico del territorio nazionaleope

La Rete sismica Mobile in telemetria satellitare (Re.Mo.Tel.)

Oggi la nuova Rete Sismica Mobile in telemetria satellitare” (Re.Mo.Tel.) è composta da nove stazioni la cui trasmissione, tramite ponte UHF (Ultra High Frequency), è equamente ripartita verso tre centri di acquisizione intermedi, detti “sottonodi”). Tali sottonodi, a loro volta tramite connessione Wi-Fi, ridirezionano il flusso dati verso un “centro stella” (detto “nodo”) dal quale, con il sistema di trasmissione satellitare Libra VSAT Nanometrics, i dati sono inviati al centro acquisizione dati della Sala Sorveglianza Sismica della sede INGV di Roma e ridondati al centro “disaster recovery” approntato presso l’Osservatorio di Grottaminarda (Sede Irpinia in provincia di Avellino). La struttura della Re.Mo.Tel. è stata ideata ed ingegnerizzata in modo da ridurre al minimo i tempi d’installazione. Il sistema è stato infatti realizzato interamente “plug and play” e di conseguenza nessuna attività di configurazione è richiesta all’operatore all’atto dell’istallazione. La Re.Mo.Tel. si basa su di un articolato sistema di trasmissione (UHF, Wi-Fi e satellitare), mediante l’impiego di diversi apparati

The RING GPS network: a research geodetic infrastructure to study plate boundary deformation in the Central Mediterranean

We present the INGV (Italian National Institute of Geophysics and Volcanology) geodetic research infrastructure and related facilities, dedicated to the observation and monitoring of current deformation of the plate boundary between Africa and Eurasia. The recent increase of continuous GPS (CGPS) stations in the Central Mediterranean plate boundary zone offers the opportunity to study in detail the present-day kinematics of this actively deforming region. For answering all the open questions related to this complex area, INGV deployed a permanent, integrated and real-time monitoring CGPS network (RING) all over Italy. The RING network (http:/ring.gm.ingv.it) is now constituted by more than 150 stations. All stations have high quality GPS monuments and most of them are co-located with broadband or very broadband seismometers and strong motion sensors. The RING CGPS sites acquire at 1Hz and 30s sampling rates (some of them acquire at 10 Hz) and are connected in real-time to the INGV acquisition centers located in Roma and Grottaminarda. Real-time GPS data are transmitted using different systems, such as satellite systems, Internet, GPRS/UMTS and wireless networks. The differentiation of data transmission type and the integration with seismic instruments makes this network one of the most innovative CGPS networks in Europe. Furthermore, the INGV data acquisition centers acquire, archive and analyze most of the Italian CGPS stations managed by regional or national data providers (such as local Authorities and nation-wide industries), integrating more than 350 stations of the CGPS scientific and commercial networks existing in the Italian region. To manage data acquisition, storage, distribution and access we developed dedicated facilities including new softwares for data acquisition and a web-based collaborative environment for management of data and metadata. The GPS analysis is carried out with the three main geodetic-quality softwares used in the GPS scientific community: Bernese GAMIT an GIPSY-OASIS. The resulting daily solutions are aligned to the ITRF2005 reference frame. Stable plate reference frames are realized by minimizing the horizontal velocities at sites on the Eurasia and Nubia plates, respectively. The different software-related solutions consistency RMS is within 0.3 mm/yr (Avallone et al., 2010). The solutions are then evaluated with regard to the numerous scientific motivations behind this presentation, ranging from the definition of strain distribution and microplate kinematics within the plate boundary, to the evaluation of tectonic strain accumulation on active faults. The RING network is strongly contributing to the definition of GPS velocity field in the Italian region, and now is able to furnish a newly and up to date view of this actively deforming part of the Nubia-Eurasia plate boundary. INGV is now aiming to make the RING (and integrated CGPS networks) data and related products publicly available for the scientific community. We believe that our network represents an important reality in the framework of the EPOS infrastructure and we strongly support the idea of an European research approach to data sharing among the scientific community. We will present (a) the current CGPS site distribution, (b) the technological description of the data acquisition, storage and distribution at INGV centers, (c) the results of CGPS data analysis, and (d) the planned data access for the scientific community.PublishedVienna, Geophysical Research Abstracts Vol. 13, EGU2011-8626, 20111.9. Rete GPS nazionale3.2. Tettonica attivaope

Genotype-phenotype correlation and functional studies in patients with cystic fibrosis bearing CFTR complex alleles

Background The effect of complex alleles in cystic fibrosis (CF) is poorly defined for the lack of functional studies. Objectives To describe the genotype-phenotype correlation and the results of either in vitro and ex vivo studies performed on nasal epithelial cells (NEC) in a cohort of patients with CF carrying cystic fibrosis transmembrane conductance regulator (CFTR) complex alleles. Methods We studied 70 homozygous, compound heterozygous or heterozygous for CFTR mutations: p. [Arg74Trp;Val201Met;Asp1270Asn], n=8; p.[Ile148Thr; Ile1023_Val1024del], n=5; p.[Arg117Leu;Leu997Phe], n=6; c.[1210-34TG[12];1210-12T[5];2930C>T], n=3; p. [Arg74Trp;Asp1270Asn], n=4; p.Asp1270Asn, n=2; p. Ile148Thr, n=6; p.Leu997Phe, n=36. In 39 patients, we analysed the CFTR gating activity on NEC in comparison with patients with CF (n=8) and carriers (n=4). Finally, we analysed in vitro the p.[Arg74Trp;Val201Met;Asp1270Asn] complex allele. Results The p.[Ile148Thr;Ile1023_Val1024del] caused severe CF in five compound heterozygous with a class I-II mutation. Their CFTR activity on NEC was comparable with patients with two class I-II mutations (mean 7.3% vs 6.9%). The p.[Arg74Trp;Asp1270Asn] and the p. Asp1270Asn have scarce functional effects, while p. [Arg74Trp;Val201Met;Asp1270Asn] caused mild CF in four of five subjects carrying a class I-II mutation in trans, or CFTR-related disorders (CFTR-RD) in three having in trans a class IV-V mutation. The p.[Arg74Trp;Val201Met; Asp1270Asn] causes significantly (pT] and a class I-II mutation had mild CF or CFTR-RD (gating activity: 18.5-19.0%). Conclusions The effect of complex alleles partially depends on the mutation in trans. Although larger studies are necessary, the CFTR activity on NEC is a rapid contributory tool to classify patients with CFTR dysfunction