A mutation in the melon Vacuolar Protein Sorting 41prevents systemic infection of Cucumber mosaic virus

[EN] In the melon exotic accession PI 161375, the gene cmv1, confers recessive resistance to Cucumber mosaic virus (CMV) strains of subgroup II. cmv1 prevents the systemic infection by restricting the virus to the bundle sheath cells and impeding viral loading to the phloem. Here we report the fine mapping and cloning of cmv1. Screening of an F2 population reduced the cmv1 region to a 132 Kb interval that includes a Vacuolar Protein Sorting 41 gene. CmVPS41 is conserved among plants, animals and yeast and is required for post-Golgi vesicle trafficking towards the vacuole. We have validated CmVPS41 as the gene responsible for the resistance, both by generating CMV susceptible transgenic melon plants, expressing the susceptible allele in the resistant cultivar and by characterizing CmVPS41 TILLING mutants with reduced susceptibility to CMV. Finally, a core collection of 52 melon accessions allowed us to identify a single amino acid substitution (L348R) as the only polymorphism associated with the resistant phenotype. CmVPS41 is the first natural recessive resistance gene found to be involved in viral transport and its cellular function suggests that CMV might use CmVPS41 for its own transport towards the phloem.The TILLING platform is supported by the Program Saclay Plant Sciences (SPS, ANR-10-LABX-40) and the European Research Council (ERC-SEXYPARTH). This work was supported by grants AGL2009-12698-C02-01 and AGL2012-40130-C02-01 from the Spanish Ministry of Science and Innovation, the Spanish Ministry of Econom and Competitiveness, through the "Severo Ochoa Programme for Centres of Excellence in R&D" 2016-2019 (SEV-2015-0533)" and the CERCA Programme/Generalitat de Catalunya.Giner, A.; Pascual, L.; Bourgeois, M.; Gyetvai, G.; Rios, P.; Picó Sirvent, MB.; Troadec, C.... (2017). 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Synthetic Culture Media Evaluated for the Detection of Coliform Bacteria in Milk, Cheese and Egg Melange

Simple synthetic culture media of liquid and solid form (X broth and X agar) were tested for selective isolation of coliform bacteria. Selectivity is based on the ability of coliform bacteria to grow when the minimal medium contains simple inorganic substances as nitrogen and carbon supply. Selectivity of the media was tested by inoculation of pure cultures of different microbes belonging to the genera of Staphylococcus, Bacillus and Pseudomonas and the family Enterobacteriaceae and was found to be complete in this range. The comparative investigation of milk, camembert cheese and egg melange samples in the traditional and new media proved good applicability of X broth and X agar for an effective and selective detection of coliform bacteria. When testing pasteurized milk samples, X agar detected coliforms in significantly higher counts than violet red-bile-lactose agar

Gentamicin sulphate permeation through porcine intestinal epithelial cell monolayer

Gentamicin is an aminoglycoside antibiotic widely used in combination with dimethyl sulphoxide (DMSO) in topical drug formulations. It is not known, however, whether DMSO can enhance the permeation of gentamicin through biological membranes, leading to oto- and nephrotoxic side effects. A simple and reliable high-performance liquid chromatographic (HPLC) method was applied for the quantitative determination of gentamicin collected from the apical and basolateral compartments of the porcine intestinal epithelial cell line IPEC-J2 cell monolayer using fluorometric derivatisation of the analyte with fluorenylmethyloxycarbonyl chloride (FMOC) prior to chromatographic run in the presence and absence of 1% DMSO. The lack of change in transepithelial electrical resistance (TER) demonstrated that gentamicin and 1% DMSO did not affect IPEC-J2 cell monolayer integrity via the disruption of cell membranes. Chromatographic data also ascertained that gentamicin penetration across the cell monolayer even in the presence of 1% DMSO was negligible at 6 h after the beginning of apical gentamicin administration. This study further indicates that the addition of this organic solvent does not increase the incidence of toxic effects related to gentamicin permeation

Transcription analysis of the myometrium of labouring and non-labouring women

An incomplete understanding of the molecular mechanisms that initiate normal human labour at term seriously hampers the development of effective ways to predict, prevent and treat disorders such as preterm labour. Appropriate analysis of large microarray experiments that compare gene expression in non-labouring and labouring gestational tissues is necessary to help bridge these gaps in our knowledge. In this work, gene expression in 48 (22 labouring, 26 non-labouring) lower-segment myometrial samples collected at Caesarean section were analysed using Illumina HT-12 v4.0 BeadChips. Normalised data were compared between labouring and non-labouring groups using traditional statistical methods and a novel network graph approach. We sought technical validation with quantitative real-time PCR, and biological replication through inverse variance-weighted meta-analysis with published microarray data. We have extended the list of genes suggested to be associated with labour: Compared to non-labouring samples, labouring samples showed apparent higher expression at 960 probes (949 genes) and apparent lower expression at 801 probes (789 genes) (absolute fold change ≥1.2, rank product percentage of false positive value (RP-PFP) <0.05). Although half of the women in the labouring group had received pharmaceutical treatment to induce or augment labour, sensitivity analysis suggested that this did not confound our results. In agreement with previous studies, functional analysis suggested that labour was characterised by an increase in the expression of inflammatory genes and network analysis suggested a strong neutrophil signature. Our analysis also suggested that labour is characterised by a decrease in the expression of muscle-specific processes, which has not been explicitly discussed previously. We validated these findings through the first formal meta-analysis of raw data from previous experiments and we hypothesise that this represents a change in the composition of myometrial tissue at labour. Further work will be necessary to reveal whether these results are solely due to leukocyte infiltration into the myometrium as a mechanism initiating labour, or in addition whether they also represent gene changes in the myocytes themselves. We have made all our data available at www.ebi.ac.uk/arrayexpress/ (accession number E-MTAB-3136) to facilitate progression of this work

Discomfort: not pain but still unpleasant feelings from the gut

Most of the factors initiating food or fluid intake have already been studied, but much less is known about those terminating ingestion. We have hypothetised that discomfort originating from the gastrointestinal system may be one of those factors. Gut distension cause pain if the intestinal volume changes but merely discomfort if only the tension of the gut wall increases. It seems that mild unpleasantness (i.e. discomfort) arising from the gut as a result of moderate (quasi-isometric) distension, among and in concordance with other factors, may significantly reduce intake and hence contribute to physiological satiety. The arising discomfort can be detected by measuring the amount and rate of the ingestion, by recording and analysing ingestive behavior by taste-aversivity and taste-reactivity tests, etc. Conclusions of all experiments point to the same direction: tension increase in the gut wall causes discomfort and results in decrease of intake, i.e. satiety

Channel-pattern adjustments and geomorphic characteristics of Elkhead Creek, Colorado, 1937-97 /

Includes bibliographical references (p. 38-39).Mode of access: Internet

Recent advances in the prevention and treatment of preterm labour: Oxytocin antagonists and the silicone (Arabin) pessary

© 2015 Cambridge University Press. Preterm birth may be spontaneous or medically indicated for maternal or fetal reasons. Around 20-25% of preterm births (PTB) follow preterm premature rupture of the membranes (PPROM), however the cause of preterm labour is often unknown. It may represent early maturation and activation of the normal labour process or it may be precipitated by pathological causes. The normal process of labour has a diurnal variation with more deliveries occurring at night. Evidence demonstrating that the diurnal variation persists in preterm deliveries suggest that at least a proportion are due to early maturation of the normal process and the logical assumption is that these may be amenable to prevention or effective treatment. Whatever the cause of preterm delivery, there appears to be a common pathway resulting in activation of inflammatory processes. It is important to distinguish the physiological and pathological causes of preterm labour and not to assume that all inflammation is pathological. The distinction is clinically important since pathological causes may be associated with an adverse intrauterine environment, which would be a contraindication to delaying delivery