A Comprehensive Analysis of Gene Expression Changes Provoked by Bacterial and Fungal Infection in C. elegans

While Caenorhabditis elegans specifically responds to infection by the up-regulation of certain genes, distinct pathogens trigger the expression of a common set of genes. We applied new methods to conduct a comprehensive and comparative study of the transcriptional response of C. elegans to bacterial and fungal infection. Using tiling arrays and/or RNA-sequencing, we have characterized the genome-wide transcriptional changes that underlie the host's response to infection by three bacterial (Serratia marcescens, Enterococcus faecalis and otorhabdus luminescens) and two fungal pathogens (Drechmeria coniospora and Harposporium sp.). We developed a flexible tool, the WormBase Converter (available at http://wormbasemanager.sourceforge.net/), to allow cross-study comparisons. The new data sets provided more extensive lists of differentially regulated genes than previous studies. Annotation analysis confirmed that genes commonly up-regulated by bacterial infections are related to stress responses. We found substantial overlaps between the genes regulated upon intestinal infection by the bacterial pathogens and Harposporium, and between those regulated by Harposporium and D. coniospora, which infects the epidermis. Among the fungus-regulated genes, there was a significant bias towards genes that are evolving rapidly and potentially encode small proteins. The results obtained using new methods reveal that the response to infection in C. elegans is determined by the nature of the pathogen, the site of infection and the physiological imbalance provoked by infection. They form the basis for future functional dissection of innate immune signaling. Finally, we also propose alternative methods to identify differentially regulated genes that take into account the greater variability in lowly expressed genes

Wood specific gravity variations within tree trunk: the case study of Legumes representatives in French Guiana

International audienceOver the past decade, much attention has been devoted to the development of forest biomass estimation methods at a stand scale, leading to the establishment of allometric models (Chave et al., 2014). These allometric equations use a unique wood specific gravity value (WSG) per species, but neglect the within tree variations of WSG found by others (Wiemann & Williamson, 1989).The main objectives of this study are (1) to illustrate the diversity of radial (from pith to bark) and longitudinal (from bottom to top) patterns of WSG variation within and between species, (2) to highlight different trends of WSG radial variations and the possible misinterpretations of these trends due to the effect of heartwood and (3) to link these variations and patterns to the successional status of the species (from pioneer to sciaphilic species).We sampled 33 small trees (10<DBH<15cm) at the Paracou field station in French Guiana, belonging to 14 Legumes species, and to different ecological groups according to light. WSG radial profiles were measured at 3 heights along the trunk, and 2 heights along the crown, of each tree.We observed different radial and longitudinal patterns of WSG variation. Pioneer and heliophilic species show both radial and longitudinal increases in WSG, while shade-tolerant and sciaphilic species show the reverse pattern. Hemi-tolerant species show an intermediate pattern, with WSG increasing radially, but decreasing or increasing longitudinally. Decreasing radial pattern in sciaphilic species is due to the presence of heartwood relatively denser than sapwood. When a corrected WSG is used, sciaphilic species show the same radial pattern as hemi-tolerant species (i.e. increasing) or no radial pattern (i.e. ‘flat’ from pith to bark).Decreasing WSG from bottom to top is a general case, excepted for species with low WSG (i.e pioneers). All studied species tend to the same range of WSG values with height (~ 0.6-0.9), supported by a higher WSG under bark within trunk.We also developed a biomass model, implemented under Xplo software (Griffon et al., 2011) to infer trunk biomass from WSG profiles, allowing comparisons of both single- and varying-WSG models.Wood specific gravity variations within tree trunk: the case study of Legumes representatives in French Guiana

Diversity of experimentation by farmers engaged in agroecology

International audienceAbstractAgroecology questions the production of generic knowledge. Rather than searching for the best practices for large-scale transfer, it would be more efficient to help farmers find their own solutions. A promising activity for farmers is experimentation because it answers their needs and helps them learn. However, how agroecological practices are tested by farmers in their own experiments is still poorly known. In this study, we examined the short-term experimental activity, i.e., experiments carried out at a yearly scale in pre-defined fields. Seventeen farmers in south eastern France were surveyed. The farmers practiced conventional or organic farming and cultivated either arable or market garden crops. Experiments on agroecological practices were characterized, located along a timeline, and discussed with them. To conduct the interviews with the farmers, each experiment was described in three stages: (1) designing the experiment, (2) managing it in real time, and (3) evaluating the results of the experiment. The data collected in the interviews were first analyzed to build a descriptive framework of farmers’ experiments, after which hierarchical cluster analysis was used to analyze the diversity of the farmers’ experiments. Here, we propose for the first time a generic framework to describe farmers’ experiments at a short time scale based on the consistency between the Design, Management, and Evaluation stages. We used the framework to characterize the diversity of farmers’ experiments and identified four clusters. The originality of this work is both building a descriptive framework resulting from in-depth analyses of farmers’ discourse and using statistical tools to identify and interpret the groups of experiments. Our results provide a better understanding of farmers’ experiments and suggest tools and methods to help them experiment, a major challenge in the promotion of a large-scale agroecological transition

Vascular network remodeling via vessel cooption, regression and growth in tumors

The transformation of the regular vasculature in normal tissue into a highly inhomogeneous tumor specific capillary network is described by a theoretical model incorporating tumor growth, vessel cooption, neo-vascularization, vessel collapse and cell death. Compartmentalization of the tumor into several regions differing in vessel density, diameter and in necrosis is observed for a wide range of parameters in agreement with the vessel morphology found in human melanoma. In accord with data for human melanoma the model predicts, that microvascular density (MVD, regarded as an important diagnostic tool in cancer treatment, does not necessarily determine the tempo of tumor progression. Instead it is suggested, that the MVD of the original tissue as well as the metabolic demand of the individual tumor cell plays the major role in the initial stages of tumor growth.Comment: 30 pages, 11 figures (higher resolution at http://www.uni-saarland.de/fak7/rieger/HOMEPAGE/BJ0.pdf

Managing uncertainty: a review of food system scenario analysis and modelling

Complex socio-ecological systems like the food system are unpredictable, especially to long-term horizons such as 2050. In order to manage this uncertainty, scenario analysis has been used in conjunction with food system models to explore plausible future outcomes. Food system scenarios use a diversity of scenario types and modelling approaches determined by the purpose of the exercise and by technical, methodological and epistemological constraints. Our case studies do not suggest Malthusian futures for a projected global population of 9 billion in 2050; but international trade will be a crucial determinant of outcomes; and the concept of sustainability across the dimensions of the food system has been inadequately explored so far. The impact of scenario analysis at a global scale could be strengthened with participatory processes involving key actors at other geographical scales. Food system models are valuable in managing existing knowledge on system behaviour and ensuring the credibility of qualitative stories but they are limited by current datasets for global crop production and trade, land use and hydrology. Climate change is likely to challenge the adaptive capacity of agricultural production and there are important knowledge gaps for modelling research to address

Canine ovariohysterectomy: a survey of surgeon concerns and surgical complications encountered by newly graduated veterinarians

The objective of this study was to document newly qualified veterinarians’ concerns and surgical complications encountered during canine ovariohysterectomy (cOVH) during the first year of general practice. A questionnaire investigating concerns about cOVH procedures was sent to all final-year veterinary students (group 1) enrolled at five UK universities. Participants were later asked to complete a similar questionnaire 6 months (group 2) and 12 months (group 3) after graduation, which involved grading their concern about different aspects of the cOVH procedure and reporting surgical complications encountered after completing three cOVHs. Responses were compared between different time points. There were 196 respondents in group 1, 55 in group 2, and 36 in group 3. Between groups 1 and 2, there was a statistically significant reduction in the respondents’ levels of concern in every aspect of cOVH (p < .05). Between groups 2 and 3, there was no statistically significant change in respondents’ levels of concern in any aspect of cOVH (pb.21). There was a significant reduction in the number of complications encountered by veterinarians in group 3 (39/102, 38.2%) compared to those in group 2 (117/206, 56.8%) (p ¼ .002). Employers should anticipate high levels of concern regarding all aspects of cOVHs in new graduates, and supervision during the first 6 months may be particularly useful

Increasing tumoral 5-fluorouracil concentrations during a 5-day continuous infusion: a microdialysis study

Purpose: Response to anticancer therapy is believed to be directly related to the concentration of the anticancer drug in the tumor itself. Assessment of intra-tumor drug pharmacokinetics can be helpful to gain more insight into mechanisms involved in the (in)sensitivity of tumors to anticancer therapy. We explored the pharmacokinetics of 5-fluorouracil in both plasma and tumor tissue during a 5-day continuous infusion of 5-fluorouracil in patients with cancer. Sampling for measurement of 5-fluorouracil in tumor tissue was performed using microdialysis. Experimental design: In seven patients with an accessible (sub)cutaneous tumor treated with a continuous 5-fluorouracil infusion, plasma and microdialysate samples from tumor and normal adipose tissue were collected over a period of 5 days. Results: For six patients, drug concentrations in both tumor tissue and plasma were available. Concentration-time curves of unbound 5-fluorouracil were lower in tumor tissue compared to the curves in plasma, but exposure ratios of tumor tissue versus plasma increased during the 5-day infusion period. The presence of circadian rhythmicity of 5-fluorouracil pharmacokinetics in the tumor itself was demonstrated as 5-fluorouracil concentrations in tumor extracellular fluid were higher during the night than during daytime. Conclusion: Microdialysis was successfully employed in patients with cancer during a continuous 5-day 5-fluorouracil infusion. Plasma and tumor pharmacokinetics of 5-fluorouracil differed substantially with increasing 5-fluorouracil concentrations in tumor over time, possibly resulting from a lowered interstitial fluid pressure by 5-fluorouracil itself. This microdialysis 5-fluorouracil model might be useful to monitor the effect of drug delivery modulating strategies in future studies

In vitro selectivity, in vivo biodistribution and tumour uptake of annexin V radiolabelled with a positron emitting radioisotope

The availability of a noninvasive method to detect and quantify apoptosis in tumours will enable tumour response to several cancer therapies to be assessed. We have synthesised two radiotracers, annexin V and the N-succinimidyl-3-iodobenzoic acid (SIB) derivative of annexin V, labelled with radio-iodine (124I and 125I) and provided proof of the concept by assessing specific binding and biodistribution of these probes to apoptotic cells and tumours. We have also assessed the tumour uptake of [124I]annexin V in a mouse model of apoptosis. RIF-1 cells induced to undergo apoptosis in vitro showed a drug concentration-dependent increased binding of [125I]annexin V and [125I]SIB–annexin V. In the same model system, there was an increase in terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL)-positive cells and a decrease in clonogenic survival. Radiotracer binding was completely inhibited by preincubation with unlabelled annexin V. In RIF-1 tumour-bearing mice, rapid distribution of [125I]SIB–annexin V-derived radioactivity to kidneys was observed and the radiotracer accumulated in urine. The binding of [125I]SIB–annexin V to RIF-1 tumours increased by 2.3-fold at 48 h after a single intraperitoneal injection of 5-fluorouracil (165 mg kg−1 body weight), compared to a 4.4-fold increase in TUNEL-positive cells measured by immunostaining. Positron emission tomography images with both radiotracers demonstrated intense localisation in the kidneys and bladder. Unlike [124I]SIB–annexin V, [124I]annexin V also showed localisation in the thyroid region presumably due to deiodination of the radiolabel. [124I]SIB–annexin V is an attractive candidate for in vivo imaging of apoptosis by PET

Treatment with Imatinib in NSCLC is associated with decrease of phosphorylated PDGFR-β and VEGF expression, decrease in interstitial fluid pressure and improvement of oxygenation

Elevated intratumoral interstitial fluid pressure (IFP) and tumour hypoxia are independent predictive factors for poor survival and poor treatment response in cancer patients. However, the relationship between IFP and tumour hypoxia has not yet been clearly established. Preclinical studies have shown that lowering IFP improves treatment response to cytotoxic therapy. Interstitial fluid pressure can be reduced by inhibition of phosphorylated platelet-derived growth factor receptor-β (p-PDGFR-β), a tyrosine kinase receptor frequently overexpressed in cancer stroma, and/or by inhibition of VEGF, a growth factor commonly overexpressed in tumours overexpressing p-PDGFR-β. We hypothesised that Imatinib, a specific PDGFR-β inhibitor will, in addition to p-PDGFR-β inhibition, downregulate VEGF, decrease IFP and improve tumour oxygenation. A549 human lung adenocarcinoma xenografts overexpressing PDGFR-β were grown in nude mice. Tumour-bearing animals were randomised to control and treatment groups (Imatinib 50 mg kg−1 via gavage for 4 days). Interstitial fluid pressure was measured in both groups before and after treatment. EF5, a hypoxia marker, was administered 3 h before being killed. Tumours were sectioned and stained for p-PDGFR-β, VEGF and EF5 binding. Stained sections were viewed with a fluorescence microscope and image analysis was performed. Imatinib treatment resulted in significant reduction of p-PDGFR-β, VEGF and IFP. Tumour oxygenation was also significantly improved. This study shows that p-PDGFR-β-overexpressing tumours can be effectively treated with Imatinib to decrease tumour IFP. Importantly, this is the first study demonstrating that Imatinib treatment improves tumour oxygenation and downregulates tumour VEGF expression