Genomic analysis of three cheese-borne pseudomonas lactis with biofilm and spoilage-associated behavior

Psychrotrophic pseudomonads cause spoilage of cold fresh cheeses and their shelf-life reduction. Three cheese-borne Pseudomonas sp., ITEM 17295, ITEM 17298, and ITEM 17299 strains, previously isolated from mozzarella cheese, revealed distinctive spoilage traits based on molecular determinants requiring further investigations. Genomic indexes (ANI, isDDH), MLST-based phylogeny of four housekeeping genes (16S rRNA, gyrB, rpoB and rpoD) and genome-based phylogeny reclassified them as Pseudomonas lactis. Each strain showed distinctive phenotypic traits at 15 and 30◦C: ITEM 17298 was the highest biofilm producer at both temperatures, whilst ITEM 17295 and ITEM 17299 showed the strongest proteolytic activity at 30◦C. A wider pattern of pigments was found for ITEM 17298, while ITEM 17295 colonies were not pigmented. Although the high genomic similarity, some relevant molecular differences supported this phenotypic diversity: ITEM 17295, producing low biofilm amount, missed the pel operon involved in EPS synthesis and the biofilm-related Toxin-Antitoxin systems (mqsR/mqsA, chpB/chpS); pvdS, required for the pyoverdine synthesis, was a truncated gene in ITEM 17295, harboring, instead, a second aprA involved in milk proteolysis. This work provided new insight into the food spoiler microbiota by identifying these mozzarella cheese spoilers as P. lactis; molecular targets to be exploited in the development of novel preservative strategies were also revealed

Diltiazem downregulates IL-12 production by human dendritic cells

It is well known that IL-12 plays a central role in the initiation and control of allogeneic immune response. It promotes the proliferation of lymphocytes and NK cells, cytotoxic activity of NK cells, and CTL. It was recently shown that IL-12 is involved in the regulation of T helper Th1-Th2 responses by exerting stimulatory effects on Th1 and inhibitory effects on Th2. This regulatory role is believed to result from the ability of IL-12 to induce IFN-γ production in activated T cells and NK cells.[1 and 2] Th1 cytokines (IL-2 and IFN-γ) promote both CTL and delayed-type hypersensitivity (DTH) responses, which are considered the principal effector mechanisms of allograft rejection. Diltiazem, a calcium channel blocker used in organ transplantation, is often included in clinical protocols in association with cyclosporin A and corticosteroids.[3] It was used initially because of its antinephrotoxic and antihypertensive effects, so that the undesirable side effects induced by immunosuppressive therapy could be reduced. We previously studied the effect of diltiazem on human mixed lymphocyte reactions (MLR) and on isolated human monocytes, showing the capacity of this drug to affect proinflammatory cytokine production.[4 and 5] Since dendritic cells (DCs) are the most effective antigen-presenting cells (APCs) to prime naive T cells, we were interested in determining the influence of diltiazem on human DCs. Human DCs generated from peripheral blood monocytes in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) have been characterised as immature DCs. To become fully potent APCs, DCs must undergo maturation induced either by a proinflammatory signal such as lipopolysaccharide (LPS) or by interaction with CD40L expressed on activated T lymphocytes. [6] The ability of mature DCs to act as potent APCs is due to their high expression of MHC and costimulatory molecules and also to their production of cytokines, especially IL-12. Therefore, we determined the effect of diltiazem on cytokine production by human DCs with a particular interest in IL-1β, IL-6, TNF-α, and IL-12 production

Synthesis and characterisation of a new benzamide-containing nitrobenzoxadiazole as a GSTP1-1 inhibitor endowed with high stability to metabolic hydrolysis

The antitumor agent 6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thio)hexan-1-ol (1) is a potent inhibitor of GSTP1-1, a glutathione S-transferase capable of inhibiting apoptosis by binding to JNK1 and TRAF2. We recently demonstrated that, unlike its parent compound, the benzoyl ester of 1 (compound 3) exhibits negligible reactivity towards GSH, and has a different mode of interaction with GSTP1-1. Unfortunately, 3 is susceptible to rapid metabolic hydrolysis. In an effort to improve the metabolic stability of 3, its ester group has been replaced by an amide, leading to N-(6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thio)hexyl)benzamide (4). Unlike 3, compound 4 was stable to human liver microsomal carboxylesterases, but retained the ability to disrupt the interaction between GSTP1-1 and TRAF2 regardless of GSH levels. Moreover, 4 exhibited both a higher stability in the presence of GSH and a greater cytotoxicity towards cultured A375 melanoma cells, in comparison with 1 and its analog 2. These findings suggest that 4 deserves further preclinical testing

Status of the Cylindical-GEM project for the KLOE-2 Inner Tracker

The status of the R&D on the Cylindrical-GEM (CGEM) detector foreseen as Inner Tracker for KLOE-2, the upgrade of the KLOE experiment at the DAFNE phi-factory, will be presented. The R&D includes several activities: i) the construction and complete characterization of the full-size CGEM prototype, equipped with 650 microns pitch 1-D longitudinal strips; ii) the study of the 2-D readout with XV patterned strips and operation in magnetic field (up to 1.5T), performed with small planar prototypes in a dedicated test at the H4-SPS beam facility; iii) the characterization of the single-mask GEM technology for the realization of large-area GEM foils.Comment: 4 pages, 10 figures, Presented at Vienna Conference on Instrumentation (Feb 15-20, 2010, Vienna, Austria). Submitted to the Proceeding

Therapeutic effect of interleukin 12 on mouse haemangiosarcomas is not associated with an increased anti-tumour cytotoxic T-lymphocyte activity.

In syngeneic mice, the H5V polyoma middle-T oncogene-transformed endothelioma cell line induces Kaposi's sarcoma-like cavernous haemangiomas that regress transiently, probably because of an anti-tumour immune response, but eventually grow progressively and kill the host. To evaluate the generation of tumour-specific cytotoxic T lymphocytes (CTLs), spleen cells of tumour-bearing mice were restimulated with irradiated H5V cells in mixed leucocyte-tumour cell cultures. Tumour-specific CTLs were demonstrable only when low numbers of H5V stimulator cells were used (<1 H5V cell per 50 splenocytes). We found that H5V cells secrete immunosuppressive mediators because CTL generation was blocked when H5V cells culture supernatants were added to allogeneic mixed leucocyte cultures. As numerous tumour-derived immunosuppressive mediators may interfere with interleukin 12 (IL-12) production, we tested whether IL-12 treatment of the tumour-bearing mice would augment their immune response and thus suppress tumour growth. Indeed, IL-12 inhibited tumour growth and prevented mortality, but did not increase anti-H5V CTL generation either in vitro or in vivo. Moreover, the anti-tumour activity in IL-12-treated mice was abrogated by anti-interferon (IFN)-gamma monoclonal antibody (MAb) co-administration. These results strongly suggest that the anti-tumour effect of IL-12 is principally mediated by IFN-gamma release that in turn blocks H5V cell proliferation and induces the release of factors that suppress angiogenesis

Effect of cosmic rays on the resonant gravitational wave detector NAUTILUS at temperature T=1.5 K

The interaction between cosmic rays and the gravitational wave bar detector NAUTILUS is experimentally studied with the aluminum bar at temperature of T=1.5 K. The results are compared with those obtained in the previous runs when the bar was at T=0.14 K. The results of the run at T = 1.5 K are in agreement with the thermo-acoustic model; no large signals at unexpected rate are noticed, unlike the data taken in the run at T = 0.14 K. The observations suggest a larger efficiency in the mechanism of conversion of the particle energy into vibrational mode energy when the aluminum bar is in the superconductive status.Comment: 7 pages, 3 figures, 2 tables. Accepted by Physics Letters

Particle acoustic detection in gravitational wave aluminum resonant antennas

The results on cosmic rays detected by the gravitational antenna NAUTILUS have motivated an experiment (RAP) based on a suspended cylindrical bar, which is made of the same aluminum alloy as NAUTILUS and is exposed to a high energy electron beam. Mechanical vibrations originate from the local thermal expansion caused by warming up due to the energy lost by particles crossing the material. The aim of the experiment is to measure the amplitude of the fundamental longitudinal vibration at different temperatures. We report on the results obtained down to a temperature of about 4 K, which agree at the level of about 10% with the predictions of the model describing the underlying physical process.Comment: RAP experiment, 16 pages, 7 figure

Search for Periodic Gravitational Wave Sources with the Explorer Detector

We have developped a procedure for the search of periodic signals in the data of gravitational wave detectors. We report here the analysis of one year of data from the resonant detector Explorer, searching for pulsars located in the Galactic Center (GC). No signals with amplitude greater than $\bar{h}= 2.9~10^{-24}$, in the range 921.32-921.38 Hz, were observed using data collected over a time period of 95.7 days, for a source located at $\alpha=17.70 \pm 0.01$ hours and $\delta=-29.00 \pm 0.05$ degrees. Our procedure can be extended for any assumed position in the sky and for a more general all-sky search, even with a frequency correction at the source due to the spin-down and Doppler effects.Comment: One zipped file (Latex+eps figures). 33 pages, 14 figures. This and related material also at http://grwav3.roma1.infn.it