Binding loci of RelA-containing nuclear factor-kappaB dimers in promoter regions of PHM1-31 myometrial smooth muscle cells.

Human parturition is associated with many pro-inflammatory mediators which are regulated by the nuclear factor-kappaB (NF-κB) family of transcription factors. In the present study, we employed a ChIP-on-chip approach to define genomic loci within chromatin of PHM1-31 myometrial cells that were occupied by RelA-containing NF-κB dimers in response to a TNF stimulation of 1 h. In TNF-stimulated PHM1-31 cells, anti-RelA serum enriched 13 300 chromatin regions; importantly, 11 110 regions were also enriched by anti-RelA antibodies in the absence of TNF. DNA sequences in these regions, from both unstimulated or TNF-stimulated PHM1-31 cultures, were associated with genic regions including IκBα, COX-2, IL6RN, Jun and KCNMB3. TNF-induced binding events at a consensus κB site numbered 1667; these were represented by 112 different instances of the consensus κB motif. Of the 1667 consensus κB motif occurrences, 770 (46.2%) were identified within intronic regions. In unstimulated PHM1-31 cells, anti-RelA-serum-enriched regions were associated with sequences corresponding to open reading frames of ion channel subunit genes including CACNB3 and KCNB1. Moreover, in unstimulated cells, the consensus κB site was identified 2116 times, being defined by 103 different sequence instances of this motif. Of these 2116 consensus κB motifs, 1089 (51.5%) were identified within intronic regions. Parallel expression array analyses in PHM1-31 cultures demonstrated that TNF stimulated a >2-fold induction in 51 genes and a fold repression of >1.5 in 18 others. We identified 14 anti-RelA-serum-enriched genomic regions that correlated with 17 TNF-inducible genes, such as COX2, Egr-1, Jun, IκBα and IL6, as well as five regions associated with TNF-mediated gene repression, including Col1A2

DJ-1 is not a deglycase and makes a modest contribution to cellular defense against methylglyoxal damage in neurons

Human DJ-1 is a cytoprotective protein whose absence causes Parkinson\u27s disease and is also associated with other diseases. DJ-1 has an established role as a redox-regulated protein that defends against oxidative stress and mitochondrial dysfunction. Multiple studies have suggested that DJ-1 is also a protein/nucleic acid deglycase that plays a key role in the repair of glycation damage caused by methylglyoxal (MG), a reactive α-keto aldehyde formed by central metabolism. Contradictory reports suggest that DJ-1 is a glyoxalase but not a deglycase and does not play a major role in glycation defense. Resolving this issue is important for understanding how DJ-1 protects cells against insults that can cause disease. We find that DJ-1 reduces levels of reversible adducts of MG with guanine and cysteine in vitro. The steady-state kinetics of DJ-1 acting on reversible hemithioacetal substrates are fitted adequately with a computational kinetic model that requires only a DJ-1 glyoxalase activity, supporting the conclusion that deglycation is an apparent rather than a true activity of DJ-1. Sensitive and quantitative isotope-dilution mass spectrometry shows that DJ-1 modestly reduces the levels of some irreversible guanine and lysine glycation products in primary and cultured neuronal cell lines and whole mouse brain, consistent with a small but measurable effect on total neuronal glycation burden. However, DJ-1 does not improve cultured cell viability in exogenous MG. In total, our results suggest that DJ-1 is not a deglycase and has only a minor role in protecting neurons against methylglyoxal toxicity

"The dirty hand in the latex glove": a study of hand hygiene compliance when gloves are worn.

Wearing of gloves reduces transmission of organisms by healthcare workers' hands but is not a substitute for hand hygiene. Results of previous studies have varied as to whether hand hygiene is worse when gloves are worn. Most studies have been small and used nonstandardized assessments of glove use and hand hygiene. We sought to observe whether gloves were worn when appropriate and whether hand hygiene compliance differed when gloves were worn

Nosocomial transmission of C. difficile in English hospitals from patients with symptomatic infection.

BACKGROUND: Recent evidence suggests that less than one-quarter of patients with symptomatic nosocomial Clostridium difficile infections (CDI) are linked to other in-patients. However, this evidence was limited to one geographic area. We aimed to investigate the level of symptomatic CDI transmission in hospitals located across England from 2008 to 2012. METHODS: A generalized additive mixed-effects Poisson model was fitted to English hospital-surveillance data. After adjusting for seasonal fluctuations and between-hospital variation in reported CDI over time, possible clustering (transmission between symptomatic in-patients) of CDI cases was identified. We hypothesised that a temporal proximity would be reflected in the degree of correlation between in-hospital CDI cases per week. This correlation was modelled through a latent autoregressive structure of order 1 (AR(1)). FINDINGS: Forty-six hospitals (33 general, seven specialist, and six teaching hospitals) located in all English regions met our criteria. In total, 12,717 CDI cases were identified; seventy-five per cent of these occurred >48 hours after admission. There were slight increases in reports during winter months. We found a low, but statistically significant, correlation between successive weekly CDI case incidences (phi = 0.029, 95%CI: 0.009-0.049). This correlation was five times stronger in a subgroup analysis restricted to teaching hospitals (phi = 0.104, 95%CI: 0.048-0.159). CONCLUSIONS: The results suggest that symptomatic patient-to-patient transmission has been a source of CDI-acquisition in English hospitals in recent years, and that this might be a more important transmission route in teaching hospitals. Nonetheless, the weak correlation indicates that, in line with recent evidence, symptomatic cases might not be the primary source of nosocomial CDI in England

Horticulture Research

Grafting is an ancient method that has been intensively used for the clonal propagation of vegetables and woody trees. Despite its importance in agriculture the physiological and molecular mechanisms underlying phenotypic changes of plants following grafting are still poorly understood. In the present study, we analyse the populations of small RNAs in homo and heterografts and take advantage of the sequence differences in the genomes of heterograft partners to analyse the possible exchange of small RNAs. We demonstrate that the type of grafting per se dramatically influences the small RNA populations independently of genotypes but also show genotype specific effects. In addition, we demonstrate that bilateral exchanges of small RNAs, mainly short interfering RNAs, may occur in heterograft with the preferential transfer of small RNAs from the scion to the rootstock. Altogether, the results suggest that small RNAs may have an important role in the phenotype modifications observed in heterografts

Moving towards grapevine genotypes better adapted to abiotic constraints

Vitis spp., both in their cultivated and wild forms, have been growing in a large diversity of environments for thousands of years. As a result, they have developed many adaptive mechanisms controlled by a range of regulatory processes. The cultivated species, Vitis vinifera, is quite well adapted to semi-arid conditions and its cultivation can be used to produce crops on marginal lands. However, this is threatened by climate change, which is associated with increased temperature and CO2 atmospheric content, changes in water availability and an increased likelihood of extreme events, such as heat waves and early spring frosts. Indirect effects of climate change on solar radiation and soil minerals are also expected. Consequently, cultivated grapevines will presumably face more abiotic constraints occurring concomitantly or successively over one or more growing cycles. In addition to climate change, worldwide viticulture must reduce the use of pesticides. Adapting to climate change and reducing pesticide use are challenging, and increase the need to create new grapevine varieties that are more resistant to diseases and better adapted to abiotic constraints. For this purpose, the adaptive mechanisms of wild and cultivated Vitis spp. must be exploited. While major advances have already been made in exploiting wild alleles for disease resistance, the polygenic nature of adaptation to abiotic factors has slowed down research progress. To tackle this limitation, ambitious integrative strategies need to be undertaken from collection and characterization of genetic resources, investigations on genetic architecture and identification of underlying genes (including those involved in epigenetic regulation), to the implementation of new breeding technologies and the development of genomic selection. An update on the state-of-the-art regarding these aspects is presented

Investigating the potential of novel non-woven fabrics for efficient pollination control in plant breeding

Internal temperature (a) and vapour pressure deficit (hPa, b) response rates to a transition from dark to light (0 to 880 umolm-2s-1) above the pollination control bags. Box plots of the average maximum temperature (c) and VPD (d) are shown for each bag type. Significant differences are denoted by different lower case letters for temperature and VPD and are based on 11 replicates.</p

Tissue-specific stilbene accumulation is an early response to wounding/grafting as revealed by using spatial and temporal metabolomics

Grafting is widely used in horticulture. Shortly after grafting, callus tissues appear at the graft interface and the vascular tissues of the scion and rootstock connect. The graft interface contains a complex mix of tissues, we hypothesised that each tissue has its own metabolic response to wounding/grafting and accumulates different metabolites at different rates. We made intact and wounded cuttings and grafts of grapevine, and then measured changes in bulk flavonoid, phenolic acid and stilbenoid concentration and used metabolite imaging to study tissue‐specific responses. We show that some metabolites rapidly accumulate in specific tissues after grafting, for example, stilbene monomers accumulate in necrotic tissues surrounding mature xylem vessels. Whereas other metabolites, such as complex stilbenes, accumulate in the same tissues at later stages. We also observe that other metabolites accumulate in the newly formed callus tissue and identify genotype‐specific responses. In addition, exogenous resveratrol application did not modify grafting success rate, potentially suggesting that the accumulation of resveratrol at the graft interface is not linked to graft union formation. The increasing concentration of complex stilbenes often occurs in response to plant stresses (via unknown mechanisms), and potentially increases antioxidant activity and antifungal capacities

Evaluation of criteria to assist the selection of good quality grafted grapevines prior to their commercialisation

The production of grafted grapevine plant material is a complex process with many steps running over at least one year, from grafting to final sorting in nurseries. To reach the market in France, grafted grapevines must meet three criteria by law: resistance to a manual graft union test (or thumb test), a minimum number of three roots and a woody, lignified stem which has grown from the bud of the scion wood of at least 2 cm long. This study aimed to evaluate the possibility of using visual criteria to select good quality grafted grapevines, without the need to do the thumb test because the thumb test is manual and therefore very subjective; the test depends on the strength applied by the person who realises it. This study was done on 22 scion/rootstock combinations with different degrees of grafting success, i.e., producing different proportions of marketable plants after one year in the nursery. The three legal criteria currently used to select marketable grafted grapevines in France as well as other external and measurable criteria such as the length of lignified stem and diameter, the number of thin and thick roots, and rootstock wood diameter were measured on the 22 scion/rootstock combinations. Variation in the values for these different criteria was observed and correlations between the criteria and the number of marketable plants were studied. This data was then analysed to determine which visible criteria contribute most to identifying marketable grafts. The percentage of marketable grafts was most strongly correlated with the thumb test and positively correlated with the length of the lignified stem. The variables with the highest predictive effect for identifying marketable plants (other than the thumb test) were the number of large roots and the length of the lignified stem. The possibility of using visible criteria to screen for good quality grafted plants is discussed, but no single, or combination of criteria, was sufficiently strongly correlated with the percentage of marketable plants to replace the thumb test