The tomato ethylene receptor LE-ETR3 (NR) is not involved in mediating ozone sensitivity: causal relationships among ethylene emission, oxidative burst and tissue damage

The causal relationships among ethylene emission, oxidative burst and tissue damage, and the temporal expression patterns of some ethylene biosynthetic and responsive genes, were examined in the Never ripe (Nr) tomato (Lycopersicon esculentum) mutant and its isogenic wild type (cv. Pearson), to investigate the role played by the ethylene receptor LE-ETR3 (NR) in mediating the plant response to ozone (O(3)). Tomato plants were used in a time-course experiment in which they were exposed to acute O(3) fumigation with 200 nl l(-1) O(3) for 4 h. The pattern of leaf lesions indicated similar sensitivities to O(3) for cv. Pearson and Nr. In both genotypes, O(3) activated a hydrogen peroxide (H(2)O(2))-dependent oxidative burst, which was also ethylene-driven in Nr leaves. Ozone induced some ethylene and jasmonate biosynthetic and inducible genes, although with different timings and to different extents in the two genotypes. The overall data indicate that Nr retains partial sensitivity to ethylene, suggesting only a marginal role of the NR receptor in mediating the complex response of tomato plants to O(3).     

Exhaled breath profiling by electronic nose enabled discrimination of allergic rhinitis and extrinsic asthma

Aim: To assess whether an e-nose could discriminate between subjects affected by allergic rhinitis with and without concomitant extrinsic asthma, as well as from healthy controls, in terms of exhaled VOC-profile.

Methods: Fourteen patients with Extrinsic Asthma and Allergic Rhinitis (AAR), 14 patients with Allergic Rhinitis without asthma (AR) and 14 healthy controls (HC) participated in a cross-sectional study. Exhaled breath was collected by a standardized method and sampled by an e-nose (Cyranose 320). Raw data were reduced by Principal component analysis and analyzed by canonical discriminant analysis. Cross-validation accuracy (CVA) and Receiver Operating Characteristic(ROC)-curves were calculated. External validation in newly recruited patients (7 AAR, 7?AR and 7 HC) was tested using the previous training model.

Results: Breathprints of patients with AR clustered from those with AAR (CVA?=?85.7%), as well as HC (CVA?=?82.1%). Breathprints from AAR were also separated from those of HC (CVA?=?75.0%). External validation confirmed the above findings.

Conclusions: An e-nose can discriminate exhaled breath from subjects with allergic rhinitis with and without extrinsic asthma, which represent two different diseases with partly overlapping features. This supports the view of using breath profiling to diagnose asthma also in patients with allergic rhinitis.     

The human melanoma associated protein melanotransferrin promotes endothelial cell migration and angiogenesis in vivo

Melanotransferrin is a member of the transferrin family, which is comprised of serum transferrin, lactoferrin and ovotransferrin, and is highly expressed on melanoma cells compared to normal melanocytes. Since melanoma is an highly vascularized tumour that expresses melanotransferrin at high levels, we tested purified recombinant melanotransferrin for its capability to induce angiogenesis in the chick chorioallantoic membrane. Macroscopic and microscopic evaluation of the vascular density demonstrated that melanotransferrin exerts an angiogenic response quantitatively similar to that elicited by fibroblast growth factor-2. Overexpression of vascular endothelial growth factor-receptor-2 was observed in newly formed vessels, suggesting that the angiogenic activity of melanotransferrin may depend on activation of endogenous vascular endothelial growth factor. In addition, when antibodies against vascular endothelial growth factor were included in the assay, the angiogenic response was inhibited by 50%. In a Boyden chamber assay purified recombinant melanotransferrin induced chemotactic migration of vascular cells, which was decreased in the the presence of anti-vascular endothelial growth factor antibodies suggesting an involvement of vascular endothelial growth factor present in endothelial cells also in this assay. However, melanotransferrin was found not to directly bind to integrin alphavbeta3 or the vascular endothelial growth factor-receptor-2 as assessed in a BlAcore assay. A possible correlation between vascularization occurring during melanoma progression and the expression of melanotransferrin and vascular endothelial growth factor was established by immunolocalization of the two factors in sections of melanoma at different clinical steps of melanoma progression. These latter data strongly imply that melanotransferrin may participate in the vascularization of solid tumours and that inhibition of melanotransferrin could form the basis for intervention in tumours which use this pathway.     

Freeze-drying duration influences the amino acid and rutin content in honeybee-collected chestnut pollen

Honeybee-collected pollen is gaining attention as functional food for human consumption, due to antiproliferative, antiallergic, antibiotic, antidiarrheic and antioxidant activities. Among the different bioactive compounds, flavonoids from bee-collected pollen are currently recognised as powerful antioxidant and antiradical molecules. Traditional conservation methods influence pollen organoleptic properties as well as the contents of nutrients and nutraceutical compounds. Here, freeze-drying (FD) was proposed as a novel conservation method, estimating its adequacy as drying process by the evaluation of changes in free and total amino acids and proline as well as in their ratios. Honeybee-collected chestnut pollen was taken into consideration and the level of rutin, as main flavonoid, was considered as marker compound highlighting the maintenance of pollen nutraceutical properties. Results showed that FD influenced rutin level, depending on the FD duration. However, the free proline to free amino acid ratio was always below 80%, and the free amino acid to total amino acid ratio remained unaltered indicating the adequacy of the FD treatment, which did not affect the nutritional value of chestnut pollen. Overall, this study shed light on the nutraceutical profile of honeybee-collected chestnut pollen, highlighting the promising potential of FD as a novel method to treat pollen for human consumption.     

Oxidative damage in human gingival fibroblasts exposed to cigarette smoke

Cigarette smoke, a complex mixture of over 7000 chemicals, contains many components capable of eliciting oxidative stress, which may induce smoking-related disorders, including oral cavity diseases. In this study, we investigated the effects of whole (mainstream) cigarette smoke on human gingival fibroblasts (HGFs). Cells were exposed to various puffs (0.5-12) of whole cigarette smoke and oxidative stress was assessed by 2',7'-dichlorofluorescein fluorescence. The extent of protein carbonylation was determined by use of 2,4-dinitrophenylhydrazine with both immunocytochemical and Western immunoblotting assays. Cigarette smoke-induced protein carbonylation exhibited a puff-dependent increase. The main carbonylated proteins were identified by means of two-dimensional electrophoresis and MALDI-TOF mass spectrometry (redox proteomics). We demonstrated that exposure of HGFs to cigarette smoke decreased cellular protein thiols and rapidly depleted intracellular glutathione (GSH), with a minimal increase in the intracellular levels of glutathione disulfide and S-glutathionylated proteins, as well as total glutathione levels. Mass spectrometric analyses showed that total GSH consumption is due to the export by the cells of GSH-acrolein and GSH-crotonaldehyde adducts. GSH depletion could be a mechanism for cigarette smoke-induced cytotoxicity and could be correlated with the reduced reparative and regenerative activity of gingival and periodontal tissues previously reported in smokers.     

Real-time PCR detection of Paenibacillus spp. in raw milk to predict shelf life performance of pasteurized fluid milk products

Psychrotolerant sporeformers, specifically Paenibacillus spp., are important spoilage bacteria for pasteurized, refrigerated foods such as fluid milk. While Paenibacillus spp. have been isolated from farm environments, raw milk, processing plant environments, and pasteurized fluid milk, no information on the number of Paenibacillus spp. that need to be present in raw milk to cause pasteurized milk spoilage was available. A real-time PCR assay targeting the 16S rRNA gene was designed to detect Paenibacillus spp. in fluid milk and to discriminate between Paenibacillus and other closely related spore-forming bacteria. Specificity was confirmed using 16 Paenibacillus and 17 Bacillus isolates. All 16 Paenibacillus isolates were detected with a mean cycle threshold (C(T)) of 19.14 ± 0.54. While 14/17 Bacillus isolates showed no signal (C(T) > 40), 3 Bacillus isolates showed very weak positive signals (C(T) = 38.66 ± 0.65). The assay provided a detection limit of approximately 3.25 × 10(1) CFU/ml using total genomic DNA extracted from raw milk samples inoculated with Paenibacillus. Application of the TaqMan PCR to colony lysates obtained from heat-treated and enriched raw milk provided fast and accurate detection of Paenibacillus. Heat-treated milk samples where Paenibacillus (≥1 CFU/ml) was detected by this colony TaqMan PCR showed high bacterial counts (>4.30 log CFU/ml) after refrigerated storage (6°C) for 21 days. We thus developed a tool for rapid detection of Paenibacillus that has the potential to identify raw milk with microbial spoilage potential as a pasteurized product.     

Activation of nervous system development genes in bone marrow derived mesenchymal stem cells following spaceflight exposure

Stalled cell division in precursor bone cells and reduced osteoblast function are considered responsible for the microgravity‐induced bone loss observed during spaceflight. However, underlying molecular mechanisms remain unraveled. Having overcome technological difficulties associated with flying cells in a space mission, we present the first report on the behavior of the potentially osteogenic murine bone marrow stromal cells (BMSC) in a 3D culture system, flown inside the KUBIK aboard space mission ISS 12S (Soyuz TMA‐8 + Increment 13) from March 30 to April 8, 2006 (experiment “Stroma‐2”). Flight 1g control cultures were performed in a centrifuge located within the payload. Ground controls were maintained on Earth in another KUBIK payload and in Petri dishes. Half of the cultures were stimulated with osteo‐inductive medium. Differences in total RNA extracted suggested that cell proliferation was inhibited in flight samples. Affymetrix technology revealed that 1,599 genes changed expression after spaceflight exposure. A decreased expression of cell‐cycle genes confirmed the inhibition of cell proliferation in space. Unexpectedly, most of the modulated expression was found in genes related to various processes of neural development, neuron morphogenesis, transmission of nerve impulse and synapse, raising the question on the lineage restriction in BMSC. J. Cell. Biochem. 111: 442–452, 2010. © 2010 Wiley‐Liss, Inc.