Alpha-defensins (α-Defs) in Crohn's disease: decrease of ileal α-Def 5 via permanent methylation and increase in plasma α-Def 1-3 concentrations offering biomarker utility.

An impaired expression of α-defensins (α-Defs) in the ileal mucosa and, conversely, increased levels in plasma, have been reported in Crohn's disease (CD). However, the specificity and correlation of these findings with the degree of inflammation are unclear. We aimed to characterize the concentration and utility of ileal and plasma α-Defs in CD and to analyse a potential epigenetic mechanism of α-Def expression. Peripheral blood samples and ileal biopsies were obtained from patients at disease onset (aCD), from those who achieved remission (iCD) and from two control groups (healthy controls and non-CD-aetiology ileitis patients). Plasma α-Defs 1-3 and 4 were detected by enzyme-linked immunosorbent assay (ELISA); α-Def 5 by immunolocalization. Methylation analysis of the α-Def 5 gene was performed using the MassARRAY EpiTYPER system. Plasma α-Defs 1-3 concentrations were significantly higher in aCD with ileal involvement (L1, L3) versus iCD or the control groups. The α-Defs 1-3 concentrations were also similar to healthy controls in patients with non-CD ileitis. There was a significant positive correlation between plasma α-Defs 1-3 levels in aCD and the endoscopic index, as well as with C-reactive protein (CRP) levels. The immunopositivity scoring showed significantly reduced α-Def 5 expression in ileal inflamed (aCD) versus non-inflamed mucosa (iCD and healthy controls). The α-Def 5 gene showed a higher methylation status in CD patients than controls, regardless of the inflammation. Plasma α-Defs 1-3 concentrations correlate with the degree of inflammation and appear to be specific biomarkers of ileal-CD at diagnosis. Ileal α-Def 5 expression is down-regulated permanently by methylation.

Distribution and seasonal variability of trace elements in atmospheric particulate in the Venice Lagoon.

Size distribution and selected element concentrations of atmospheric particulate matter (PM) were investigated in the Venice Lagoon, at three sites characterised by different anthropogenic influence. The PM(10) samples were collected in six size fractions (10-7.2, 7.2-3.0, 3.0-1.5, 1.5-0.95; 0.95-0.49 and <0.49 µm) with high volume cascade impactors, and the concentration of 17 elements (Al, As, Ca, Cd, Co, Cu, Fe, K, Li, Mg, Mn, Na, Ni, Pb, Sr, V, Zn) was determined by inductively coupled plasma quadrupole mass spectroscopy. More than 1 year of sampling activities allowed the examination of seasonal variability in size distribution of atmospheric particulates and element contents for each site. At all the stations, particles with an aerodynamic diameter <3 µm were predominant, thus accounting for more than 78% of the total aerosol mass concentration. The highest PM(10) concentrations for almost all elements were found at the site which is more influenced by industrial and urban emissions. Similarity in size distribution of elements at all sites allowed the identification of three main behavioural types: (a) elements found mainly within coarse particles (Ca, Mg, Na, Sr); (b) elements found mainly within fine particles (As, Cd, Ni, Pb, V) and (c) elements with several modes spread throughout the entire size range (Co, Cu, Fe, K, Zn, Mn). Factor Analysis was performed on aerosol data separately identified as fine and coarse types in order to examine the relationships between the inorganic elements and to identify their origin. Multivariate statistical analysis and assessment of similarity in the size distribution led to similar conclusions on the sources.

Increased lung neutrophil apoptosis and inflammation resolution in nonresponding pneumonia.

Neutrophil activation state and its relationship with an inflammatory environment in community-acquired pneumonia (CAP) remain insufficiently elucidated. We aimed to evaluate the neutrophil apoptosis and cytokine pattern in CAP patients after 72 h of treatment, and their impact on infection resolution. Apoptosis of blood and bronchoalveolar lavage (BAL) neutrophils was measured in nonresponding CAP (NCAP), in responding CAP (blood only) and in patients without infection (control). Pro-inflammatory (interleukin (IL)-6, IL-8) and anti-inflammatory (IL-10) cytokines were measured. Main outcomes were clinical stability and days of hospitalisation. Basal neutrophil apoptosis was higher in the BAL and blood of NCAP, whereas spontaneous apoptosis (after 24 h culture) was lower. Cytokines in NCAP were higher than in responding CAP and control: IL-6 was increased in BAL and blood, IL-8 in BAL and IL-10 in blood. An increased basal apoptosis (≥20%) in BAL of NCAP was associated with lower systemic IL-10 (p<0.01), earlier clinical stability (p=0.05) and shorter hospital stay (p=0.02). A significant correlation was found for systemic IL-6 and IL-10 with days to reach stability and length of stay. After 72 h of treatment, an increased basal alveolar neutrophil apoptosis might contribute to downregulation of inflammation and to faster clinical stability.

Air--sea gaseous exchange of PCB at the Venice lagoon (Italy).

Water bodies are important storage media for persistent organic pollutants (POPs) such as polychlorinated biphenyls (PCBs) and this function is increased in coastal regions because their inputs are higher than those to the open sea. The air-water interface is extensively involved with the global cycling of PCBs because it is the place where they accumulate due to depositional processes and where they may be emitted by gaseous exchange. In this work the parallel collection of air, microlayer and sub-superficial water samples was performed in July 2005 at a site in the Venice lagoon to evaluate the summer gaseous flux of PCBs. The total concentration of PCBs (sum of 118 congeners) in air varies from 87 to 273 pg m(-3), whereas in the operationally defined dissolved phase of microlayer and sub-superficial water samples it varies from 159 to 391 pg L(-1). No significant enrichment of dissolved PCB into the microlayer has been observed, although a preferential accumulation of most hydrophobic congeners occurs. Due to this behaviour, we believe that the modified two-layer model was the most suitable approach for the evaluation of the flux at the air-sea interface, because it takes into account the influence of the microlayer. From its application it appears that PCB volatilize from the lagoon waters with a net flux varying from 58 to 195 ng m(-2)d(-1) (uncertainty: +/-50-64%) due to the strong influence of wind speed. This flux is greater than those reported in the literature for the atmospheric deposition and rivers input and reveals that PCB are actively emitted from the Venice lagoon in summer months.

Polychlorinated biphenyls in sediments of the Tam Giang-Cau Hai Lagoon, Central Vietnam.

The Tam Giang-Cau Hai Lagoon, the largest in south east Asia, suffers from a great anthropogenic pressure and appears subject to a process of progressive environmental deterioration. To establish causes, history and trends of lagoon contamination, sediment samples representing three major parts of the systems were sampled and analysed for porosity, organic carbon, total nitrogen, delta(13)C, grain size, radiotracers and PCBs. PCB surficial concentrations range from 10.2 to 24.5 microg kg(-1), and maximum values are close to NOAA ERL guidelines. The downcore distribution of PCBs shows two well defined peaks in the core from Tam Giang (25.5 and 16.2 microg kg(-1)), whereas the sediment from Cau Hai is characterised by a maximum at the surface. Therefore, present trends indicate that contamination is still increasing or slightly decreasing. The prevailing congeners are 3-CB and 4-CB, hence recent PCBs appear to be mostly originated by Aroclor 1016- and 1242-like mixtures, with some samples influenced also by Aroclor 1248. Present sources seem to be similar everywhere and probably they are widespread all over the territory of the Thua Thien-Hue province.

PCBs and PAHs in sea-surface microlayer and sub-surface water samples of the Venice Lagoon (Italy).

Polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) are two classes of micropollutants intensively monitored and regulated due to their toxicity, persistency and wide diffusion. Their concentrations have been investigated in sea-microlayer (SML) and sub-surface water (SSW) samples collected at two sites of the Venice Lagoon, a fragile ecosystem highly influenced by industrial and anthropogenic emissions. The total sigmaPCB concentration varies from 0.45 ng/l to 2.1 ng/l in SSW while a clear enrichment is observed in the SML, where it ranges from 1.2 ng/l to 10.5 ng/l. The total sigmaPAH concentration shows marked differences between the two stations and varies from 12.4 ng/l to 266.8 ng/l in SSW; in SML it is more uniform and ranges from 19.6 ng/l to 178.9 ng/l. The enrichment factors are not larger than 1 for both pollutants in the 'dissolved' phase, while they are most significant for the 'particulate' phase (sigmaPCB: 5-9; sigmaPAH: 4-14).

Trace metals in aerosol at Terra Nova Bay, Antarctica.

Atmospheric particulate with an aerodynamic diameter <10 microm (PM10) was sampled continuously during the austral summers of 2000-2001 and 2001-2002 at a coastal site near to the Italian base of Terra Nova, Antarctica. Li, Pb, Cd, U, Ba, Bi, Cs, Rb, Tl, Sr, Al, V, Fe, Cu, Mn, Zn, Co, Ag were determined by inductively coupled sector field mass spectroscopy (ICP-SFMS) after sample digestion by a combination of HF, HNO3, and H2O2 in ultraclean conditions. Quality control of the analytical procedure was carried out by blank control, by evaluating the limits of detection, recoveries and repeatability. Concentrations found are extremely low for most metals, confirming the high purity of Antarctic aerosol. Principal Component Analysis (PCA) highlights high correlations among Pb, Cr, Bi, Cu and Zn concentration values and among Li, U, Ba, Cs, Rb, Al, V, Fe, Mn, Co concentration values permitting the identification of two principal source groups, namely crustal dust and human emission activities. Elements of anthropogenic origins (Pb, Cr, Cu, Zn) were highly enriched with respect to their crustal composition.

Determination of polychlorobiphenyl congeners (PCBs) in the surface water of the Venice lagoon.

The purpose of this study is to evaluate the degree of PCB contamination of Venice lagoon water. The PCB determination was made on the filtrate ("dissolved PCBs") and on the particulate collected on the filters ("particulate PCBs") by continuous liquid-liquid extraction and sonication, respectively, and by HRGC-LRMS analysis. An estimation of the procedural and "working" blanks was also made. Water samples were collected at six sites, selected to represent all the pollution situations present in the lagoon, and at a site in the Adriatic Sea. The range of the total sum ("dissolved plus particulate") of 49 congener concentrations falls between 355 and 1868 pgl(-1); the "dissolved" fraction concentrations (250-792 pgl(-1)) are greater at six sites than that of the "particulate" fraction (105-1273 pgl(-1)). Chemometric analyses suggest that the PCB congener pattern in the Venice Lagoon is different from that in the Adriatic Sea.

Stability of PEI-DNA and DOTAP-DNA complexes: effect of alkaline pH, heparin and serum.

DNA complexes formed with nonviral vectors such as polyethylenimine (PEI) or 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) are widely used in gene therapy. These complexes prevent the interaction of DNA with the fluorescent probes usually employed to quantify DNA. We thus studied the procedures for DNA quantification from DNA complexes as well as their stability in the presence of DNase or mouse, rat and human sera. Release of the DNA from its complexes was accomplished by increasing the pH of the medium (from 7.3 to 13.4) or by adding heparin. The stability against degradation was tested in vitro, by incubating the complexes at 37 degrees C in the presence of DNase I and sera from the three species. Both high pH and heparin were able to release DNA from its complexes. Naked DNA formed aggregates with serum proteins that delayed electrophoresis migration, and this effect was reversed in the presence of heparin. However, these aggregates did not protect DNA from digestion by serum DNase, and the DNA digesting ability of serum was: mouse>rat>human. The DNA from the complexes was resistant to degradation by DNase I, although a low proportion of DNA from the complexes was partially digested, as determined by electrophoresis. In contrast, PEI-DNA and DOTAP-DNA complexes were stable in the presence of all sera. Heparin and high pH release DNA from its complexes. The order of DNA degradation is: mouse>rat>human, but DOTAP and PEI avoid degradation of DNA by serum compounds.

Vacunas celulares en el melanoma obtenidas mediante terapia génica: experiencia y perspectivas / Melanorna cell vaccines obtained by gene therapy: experience and prospects

La transferencia a los células tumorales del gen que codifica la citocina GM-CSF, induce un reclutamiento local de células inflamatorias cuyos efectos derivan tanto en una activación efectiva de la respuesta inmune antitumoral como en una inhibición del crecimiento y lo progresión del tumor. La utilización de un modelo de vacunación con líneas celulares de melanoma (HFH18 y B16) se basa en la demostrada capacidad inmunogénica que presentan estas líneas celulares, y que puede ser incrementada transfectando dichas células con el gen que codifica la citocina GM-CSF. En este sentido, hemos observado que el transplante a ratones singénicos de clones de melanoma HFH18 modificados genéticamente para producir GM-CSF, incremento significativamente (40-60 por ciento) lo supervivencia de los animales. Ello se acompaño de un aumento en el lugar de administración del número de granulocitos, macráfagos, eosináfilos, linfocitos e histiocitos, siendo los linfocitos CD4+ y las células dendríticas los células predominantes. Así mismo, utilizando clones derivados de la línea celular B16, hemos comprobado que los clones modificados con el gen del GM-CSF (pero no IL-4) inducen una protección máxima, con el 100 por ciento de supervivencia (AU)

Determination of polychlorobiphenyls in Venice Lagoon sediments.

Polychlorinated biphenyl (PCBs) congeners were measured in 14 sediment samples collected at five sites in the Venice Lagoon selected on the basis of a supposed diversity of inputs and of pollution levels. The PCBs were determined after a sonication assisted extraction by gas chromatography mass spectrometry (GCMS). The sum of the congener concentrations determined in the samples range from 4.05 to 239.15 ng/g on a dry weight basis. The concentration values are very variable both comparing the sites and comparing different samples from the same site and can be connected to the sand percentage in the sediment samples. The correlation matrix obtained from data matrix using the congener concentrations as variables shows high values, mostly between 0.964 and 1, suggesting that the pattern of the PCB congeners and the ratios among the concentrations in Venice Lagoon sediments are more or less constant; the samples differ only in the concentration level.