Identification and characterization of a new soybean promoter induced by Phakopsora pachyrhizi, the causal agent of Asian soybean rust.

BACKGROUND: Phakopsora pachyrhizi is a biotrophic fungal pathogen responsible for the Asian soybean rust disease causing important yield losses in tropical and subtropical soybean-producing countries. P. pachyrhizi triggers important transcriptional changes in soybean plants during infection, with several hundreds of genes being either up- or downregulated. RESULTS: Based on published transcriptomic data, we identified a predicted chitinase gene, referred to as GmCHIT1, that was upregulated in the first hours of infection. We first confirmed this early induction and showed that this gene was expressed as early as 8 h after P. pachyrhizi inoculation. To investigate the promoter of GmCHIT1, transgenic soybean plants expressing the green fluorescence protein (GFP) under the control of the GmCHIT1 promoter were generated. Following inoculation of these transgenic plants with P. pachyrhizi, GFP fluorescence was detected in a limited area located around appressoria, the fungal penetration structures. Fluorescence was also observed after mechanical wounding whereas no variation in fluorescence of pGmCHIT1:GFP transgenic plants was detected after a treatment with an ethylene precursor or a methyl jasmonate analogue. CONCLUSION: We identified a soybean chitinase promoter exhibiting an early induction by P. pachyrhizi located in the first infected soybean leaf cells. Our results on the induction of GmCHIT1 promoter by P. pachyrhizi contribute to the identification of a new pathogen inducible promoter in soybean and beyond to the development of a strategy for the Asian soybean rust disease control using biotechnological approaches.

Endothelin receptors blockade blunts hypoxia-induced increase in PAP in humans.

BACKGROUND: Activation of the endothelin-1 (ET-1) pathway may be involved in hypoxia-induced pulmonary vasoconstriction, increase in pulmonary pressure and high altitude pulmonary oedema. Thus, we investigated the effect of the ETA/ETB receptor antagonist, bosentan, on pulmonary artery systolic pressure (PASP) in healthy subjects (n = 10). DESIGN: We used a double-blind, placebo-controlled, randomized, cross-over design to study the effects of a single oral dose of bosentan (250 mg) on PASP after 90-min-exposure to normobaric hypoxia (FiO(2) = 0.12). We measured PASP and cardiac output by echocardiography, systolic arterial blood pressure, arterial O(2) saturation (SaO(2)), and blood gases at rest and during a sub-maximal exercise. RESULTS: PASP in normoxia at rest was 23.5 +/- 2.7 and during exercise 39.8 +/- 11.6 mmHg (P < 0.0001). During the placebo period, hypoxia induced a significant decrease in SaO(2), PaO(2) and PCO(2) and increase in pH. PASP at rest increased significantly: 32.1 +/- 3.5 mmHg (P < 0.001 vs. normoxia). Bosentan significantly blunted the hypoxia-induced increase in PASP: bosentan: 27.0 +/- 3.3 mmHg, P = 0.002 vs. placebo at rest, but not during exercise: bosentan 39.8 +/- 11.6 vs. placebo 43.0 +/- 8.5 mmHg, ns. Bosentan had no effect on the hypoxia-induced changes in blood gases, or on cardiac output and systolic arterial blood pressure, which were not modified by hypoxia. CONCLUSION: A single oral dose of bosentan blunted an acute hypoxia-induced increase in PASP in healthy subjects, without altering cardiac output or systemic blood pressure.

Bactericidal efficacy of a 1.5min surgical hand-rubbing protocol under in-use conditions.

In healthy volunteers, surgical hand rubbing with Sterillium for 1.5min has been shown to be as effective as a 3min procedure. The aim of this study was to assess whether this result was reproducible under in-use conditions. During nine weeks in the ambulatory surgery theatre of a 750-bed tertiary care university hospital, the two surgical hand-rubbing procedures were compared with each other, and with a hand-scrubbing procedure using a povidone-iodine (4%) scrub prior to and after 25 different surgical operations for each. Imprints of the surgeon's dominant hand were taken on culture plates before and within 1min following the end of the hand-rubbing/scrubbing procedures (immediate effect) and at the end of surgery (sustained effect). Plates were incubated aerobically at 37 degrees C for 48h. Colonies were counted at 24h and 48h. Results were expressed as the number of colony-forming units per hand. No significant difference in baseline hand bacterial load was found before the hand-rubbing/scrubbing procedures among the three groups (P=0.19). With respect to immediate and sustained antimicrobial effects, a significantly greater reduction in microbial loads on the hands was achieved with the 3min hand-rubbing protocol as opposed to hand-scrubbing protocol (P=0.04 and P=0.02, respectively), but there was no difference between the reductions obtained with 1.5 and 3min rubbing protocols (P=0.41 and P=0.36, respectively). Surgical hand rubbing with Sterillium using a 1.5min protocol should be considered as an attractive alternative method for surgical hand disinfection.

Soluble interleukin-2 receptor and metalloproteinase-9 expression in head and neck cancer: prognostic value and analysis of their relationships.

In a series of 84 head and neck patients, a statistically significant correlation was observed between high serum soluble interleukin (IL)-2 receptor alpha (sIL-2Ralpha) (P = 0.034) and metalloproteinase-9 (MMP-9) concentrations (P = 0.036) at diagnosis and a shorter survival of these patients. As MMP-9 has been shown to mediate cleavage of IL-2Ralpha (CD25) by preactivated T cells, we looked for a relationship between MMP-9 expression and soluble IL-2Ralpha serum concentrations in these cancer patients. We did not find any correlation between intratumoral expression of MMP-9 or serum MMP-9 concentrations and serum sIL-2Ralpha levels. These results led us to reassess the role of MMP-9 in the release of sIL-2Ralpha. Treatment of Kit225 leukaemic cells with recombinant MMP-9 slightly decreased membrane CD25 expression and was associated with an increased concentration of sIL-2Ralpha in the supernatants. However, using a selective inhibitor of MMP-9 we did not succeed in specifically inhibiting the release of sIL-2Ralpha by the Kit225 cell line or by phytohaemagglutinin (PHA)-activated peripheral blood mononuclear cells. In addition, in a preclinical mouse model, basal serum sIL-2Ralpha concentrations and sIL-2Ralpha production by activated cells were not altered in MMP-9-deficient mice compared to wild-type mice. Interestingly, a broad spectrum metalloproteinase inhibitor inhibited the release of sIL-2Ralpha by PHA-activated peripheral blood mononuclear cells, suggesting that in contrast with current views concerning the major role of MMP-9 in the cleavage of membrane IL-2Ralpha, other proteases are involved in the shedding of sIL-2Ralpha. MMP-9 and sIL-2Ralpha appear therefore as independent prognostic markers in head and neck cancers.

Evaluation of a new disinfection procedure for ultrasound probes using ultraviolet light.

Following 183 ultrasound examinations, a randomized trial was conducted to compare three procedures for disinfection of probes under routine conditions: dry wiping with a soft, dry, non-sterile paper towel, antiseptic wiping with a towel impregnated with disinfectant spray and dry wiping followed by a 10 min ultraviolet C (UVC) cycle in a disinfection chamber. After ultrasonography, swabs were taken from transducer heads before and after cleaning and streaked onto plates that were then cultured. The number of colonies per plate was counted and organisms identified. The median microbial reduction was 100% for UVC, 98.4% for antiseptic wiping and 87.5% for dry wiping (P<0.001). The percentage of negative specimens was 88% for UVC, 16% for antiseptic wiping and 4% for dry wiping (P<0.0001). Microbial flora was isolated from 12 probes (6.6%) before cleaning, whereas specimens obtained after cleaning contained no pathogens except in one case after antiseptic wiping. UVC disinfection of ultrasound probe may provide a useful method for reducing the bacterial load under routine conditions.