Search

© 2019 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine. Background: Heart disease is an important cause of morbidity and mortality in cats, but there is limited evidence of the benefit of any medication. Hypothesis: The angiotensin-converting enzyme inhibitor benazepril would delay the time to treatment failure in cats with heart disease of various etiologies. Animals: One hundred fifty-one client-owned cats. Methods: Cats with heart disease, confirmed by echocardiography, with or without clinical signs of congestive heart failure, were recruited between 2002 and 2005 and randomized to benazepril or placebo in a prospective, multicenter, parallel-group, blinded clinical trial. Benazepril (0.5-1.0 mg/kg) or placebo was administered PO once daily for up to 2 years. The primary endpoint was treatment failure. Analyses were conducted separately for all-cause treatment failure (main analysis) and heart disease-related treatment failure (supportive analysis). Results: No benefit of benazepril versus placebo was detected for time to all-cause treatment failure (P =.42) or time to treatment failure related to heart disease (P =.21). Hazard ratios (95% confidence interval [CI]) from multivariate analysis for benazepril compared with placebo were 1.00 (0.57-1.74) for all-cause failure, and 0.99 (0.50-1.94) for forward selection and 0.93 (0.48-1.81) for bidirectional selection models for heart disease-related failure. There were no significant differences between groups over time after administration of the test articles in left atrium diameter, left ventricle wall thickness, quality of life scores, adverse events, or plasma biochemistry or hematology variables. Conclusions and Clinical Relevance: Benazepril was tolerated well in cats with heart disease, but no evidence of benefit was detected.

New biotechnologies such as assisted conception are socially embedded artefacts that raise context‐specific ethical, moral and social anxieties. In contexts where the regulations of these profitable developments are limited or ambiguous, and competition between private facilities is high, individual doctors become morally and socially responsible for determining the parameters of administering such therapies. Ethnographic research at two private fertility centres in Colombia reveals that doctors do not determine boundaries based on monetary gain but rather personal morals, social norms and professional obligations. Medical professionals hold diverse perceptions of assisted conception, and often struggle to make decisions regarding who should access such therapies, who are ideal gamete donors and the fate of extra embryos. The complexity of these perceptions applied in a context of limited regulation and the competition of private medicine impacts the praxis of assisted conception. As doctors determine the boundaries of their practice they not only create variation between clinical practices, but also make moral decisions regarding who should be parents, how families should be formed and the significance of embryos. Thus, in navigating their everyday practices, doctors also shape the social world.

Metabarcoding of the 16S rRNA gene is commonly used to characterize microbial communities, by estimating the relative abundance of microbes. Here, we present a method to retrieve the concentrations of the 16S rRNA gene per gram of any environmental sample using a synthetic standard in minuscule amounts (100 ppm to 1% of the 16S rRNA sequences) that is added to the sample before DNA extraction and quantified by two quantitative polymerase chain reaction (qPCR) reactions. This allows normalizing by the initial microbial density, taking into account the DNA recovery yield. We quantified the internal standard and the total load of 16S rRNA genes by qPCR. The qPCR for the latter uses the exact same primers as those used for Illumina sequencing of the V3‐V4 hypervariable regions of the 16S rRNA gene to increase accuracy. We are able to calculate the absolute concentration of the species per gram of sample, taking into account the DNA recovery yield. This is crucial for an accurate estimate as the yield varied between 40% and 84%. This method avoids sacrificing a high proportion of the sequencing effort to quantify the internal standard. If sacrificing a part of the sequencing effort to the internal standard is acceptable, we however recommend that the internal standard accounts for 30% of the environmental 16S rRNA genes to avoid the PCR bias associated with rare phylotypes. The method proposed here was tested on a feces sample but can be applied more broadly on any environmental sample. This method offers a real improvement of metabarcoding of microbial communities since it makes the method quantitative with limited efforts., Here, we present a spike‐and‐recovery method to get quantitative estimates from 16S rRNA surveys. The method relies on adding an artificial strand of DNA to the lysis buffer before the DNA extraction and measuring its recovery either by direct sequencing or by quantitative polymerase chain reaction (qPCR). The low detection limit achieved by qPCR allows to add minute amounts of the internal standard so that the sequencing effort is focused on the unknown sequences.

Facing up to the grand challenges posed to society today requires a policy that counts the cost of environmental damage, such as carbon emissions and air pollution. Technologies have arrived to address climate mitigation, but relatively few of these are biotechnologies. Biotechnologies in environmental applications suffer a variety of inhibitors – political, social and technical, and yet the potential cannot be denied. The greatest technical promise for future biotechnology mobilisation may be the standardisation of engineering biology that allows more rapid and less expensive reduction to practice. However, decades of metabolic engineering for bio-based chemicals and materials have brought many research successes but few commercial-scale products. To address this gap between laboratory and market, new models of R&D&I may be needed to speed up the process. In past, haste has not mattered. For the proposed generation and those that follow, there is a need for policy makers to abandon this complacency as recent evidence is showing that time is running out to keep global warming within internationally agreed limits.

édition online; Bread wheat derives from a grass ancestor structured in 7 protochromosomes followed by a paleotetraploidization to reach a 12 chromosomes intermediate and a neohexaploidization (involving subgenomes A, B and D) event that finally shaped the 21 modern chromosomes. Insights into wheat syntenome in sequencing COS (Conserved Orthologous Set) genes unravelled differences in genomic structure (such as gene conservation and diversity) and genetical landscape (such as recombination pattern) between ancestral as well as recent duplicated blocks. Contrasted evolutionary plasticity is observed where the B subgenome appears more sensitive (i.e. plastic) in contrast to A as dominant (i.e. stable) in response to the neotetraploidization and D subgenome as supradominant (i.e. pivotal) in response to the neohexaploidization event. Finally, the wheat syntenome, delivered through a public web interface PlantSyntenyViewer at http://urgi.versailles.inra.fr/synteny-wheat, can be considered as a guide for accelerated dissection of major agronomical traits in wheat. This article is protected by copyright. All rights reserved.

International audience; A global DNA methylation and proteomics approach was used to investigate somatic embryo maturation in hybrid larch. Each developmental step during somatic embryogenesis was associated with a distinct and significantly different global DNA methylation level: from 45.8% mC for undifferentiated somatic embryos (1-week proliferation) to 61.5% mC for immature somatic embryos (1-week maturation), while maturation was associated with a decrease in DNA methylation to 53.4% for mature cotyledonary somatic embryos (8-weeks maturation). The presence of 5-azacytidine (hypo-methylating agent) or hydroxyurea (hyper-methylating agent) in the maturation medium altered the global DNA methylation status of the embryogenic cultures, and significantly reduced both their relative growth rate and embryogenic potential, suggesting an important role for DNA methylation in embryogenesis. Maturation was also assessed by examining changes in the total protein profile. Storage proteins, identified as legumin- and vicilin-like, appeared at the precotyledonary stage. In the proteomic study, total soluble proteins were extracted from embryos after 1 and 8 weeks of maturation, and separated by two-dimensional gel electrophoresis. There were 147 spots which showed significant differences between the stages of maturation; they were found to be involved mainly in primary metabolism and the stabilization of the resulting metabolites. This indicated that the somatic embryo was still metabolically active at 8 weeks of maturation. This is the first report of analyses of global DNA methylation (including the effects of hyper- and hypo-treatments) and proteome during somatic embryogenesis in hybrid larch, and thus provides novel insights into maturation of conifer somatic embryos.

Correspondance: chervin@ensat.fr; International audience; Grape is considered as a non-climacteric fruit, the maturation of which is independent of ethylene. However, previous work had shown that ethylene is capable of affecting the physiological processes during maturation of grape berries. Experiments were designed to screen the gene pool affected by ethylene at the ripening inception in Cabernet Sauvignon berries. The results showed that only 73 of 14 562 genes of microarray slides were significantly modulated by a 24-h ethylene treatment (4 mu l l(-1)), performed 8 weeks after flowering. The study then focused on accumulation of several mRNAs affected by ethylene in relation to the berry size. Indeed, we observed that ethylene application at veraison led to a berry diameter increase. This increase is mainly because of sap intake and cell wall modifications, enabling cell elongation. This was related to changes in the expression pattern of many genes, classified in two groups: (1) 'water exchange' genes: various aquaporins (AQUA) and (2) 'cell wall structure' genes: polygalacturonases, xyloglucan endotransglucosylases (XTH), pectin methyl esterases, cellulose synthases and expansins. The expression patterns were followed either along berry development or in three berry tissues (peel, pulp and seeds). Ethylene stimulates the accumulation of most gene transcripts in 1 h, and in several parts of the berry, this stimulation may last for 24 h in some cases. One XTH and one AQUA seem to be good candidates to explain the ethylene-induced berry expansion. This work brings more clues about the ethylene involvement in the development and ripening of grape berries

Antisense ACC-oxidaseCantaloupeCharentaismelons, unsuitable for autocatalytic ethylene production, were harvested at different ages and exposed to a standard post-harvest manipulation to reproduce the handling chain that occurs from harvesting to retail sale. The stages were as follows: storage in a chamber at 2 ◦C during 2 or 3weeks, second storage at 8 ◦C for 4 days, treatment with 50 ppm ethylene for 2 days and, finally, maintenance at 20 ◦C for four more days. The date of harvest and the different periods of storage at 2 ◦C assayed did not affect the final de-greening of the rind, while the change in pulp colour depended on the harvest date. Storage conditions also influenced the texture of the pulp and acidity. Weight loss was dependent mainly on date of harvest, like soluble solids content, although the period of storage at 2 ◦C influenced too. The impact of cold on the appearance of the fruits is discussed.

International audience; Gene expression profiling holds tremendous promise for dissecting the regulatory mechanisms and transcriptional networks that underlie biological processes. Here we provide details of approaches used by others and ourselves for gene expression profiling in plants with emphasis on cDNA microarrays and discussion of both experimental design and downstream analysis. We focus on methods and techniques emphasizing fabrication of cDNA microarrays, fluorescent labeling, cDNA hybridization, experimental design, and data processing. We include specific examples that demonstrate how this technology can be used to further our understanding of plant physiology and development (specifically fruit development and ripening) and for comparative genomics by comparing transcriptome activity in tomato and pepper fruit.

Interspecific hybridizations were performed between Pelargonium x hortorum (subgenus Ciconium) and a yellow-flowered species, Pelargonium quinquelobatum (subgenus Eumorpha). Hybrids, using embryo rescue. were obtained only when P. quinquelobatum was used as the female parent. A polymerase chain reaction (PCR) cloned Pelargonium chs gene fragment was used as probe to characterize hybrids by restriction fragment length polymorphism (RFLP) analysis. Chromatographic flavonoid analysis showed that hybrids could synthesize both flavonol glycosides and anthocyanin in the petals which were pale yellow to pale pink from the proximal to the distal part.

Two genes (CM-AAT1 and CM-AAT2) with strong sequence homology (87% identity at the protein level) putatively involved in the formation of aroma volatile esters have been isolated from Charentais melon fruit.They belong to a large and highly divergent family of multifunctional plant acyl-transferases and show at most 21% identity to the only other fruit acyl-transferase characterized so far in strawberry.RT-PCR studies indicated that both genes were specifically expressed in fruit at increasing rates in the early and mid phases of ripening.Expression was severely reduced in ethylene-suppressed antisense ACC oxidase (AS) fruit and in wild-type (WT) fruit treated with the ethylene antagonist 1-MCP.Cloning of the two genes in yeast revealed that the CM-AAT1 protein exhibited alcohol acyl-transferase activity while no such activity could be detected for CM-AAT2 despite the strong homology between the two sequences.CM-AAT 1 was capable of producing esters from a wide range of combinations of alcohols and acyl-CoAs.The higher the carbon chain of aliphatic alcohols, the higher the activity.Branched alcohols were esterified at differential rates depending on the position of the methyl group and the nature of the acyl donor. Phenyl and benzoyl alcohols were also good substrates, but activity varied with the position and size of the aromatic residue.The cis/trans configuration influenced activity either positively (2-hexenol) or negatively (3-hexenol).Because ripening melons evolve the whole range of esters generated by the recombinant CM-AAT1 protein, we conclude that CM-AAT1 plays a major role in aroma volatiles formation in the melon.