Assessment of Transformed Properties In Vitro and of Tumorigenicity In Vivo in Primary Keratinocytes Cultured for Epidermal Sheet Transplantation

Epidermal keratinocytes are used as a cell source for autologous and allogenic cell transplant therapy for skin burns. The question addressed here is to determine whether the culture process may induce cellular, molecular, or genetic alterations that might increase the risk of cellular transformation. Keratinocytes from four different human donors were investigated for molecular and cellular parameters indicative of transformation status, including (i) karyotype, (ii) telomere length, (iii) proliferation rate, (iv) epithelial-mesenchymal transition, (v) anchorage-independent growth potential, and (vi) tumorigenicity in nude mice. Results show that, despite increased cell survival in one keratinocyte strain, none of the cultures displayed characteristics of cell transformations, implying that the culture protocol does not generate artefacts leading to the selection of transformed cells. We conclude that the current protocol does not result in an increased risk of tumorigenicity of transplanted cells

FREE ORAL COMMUNICATIONS 2: ALCOHOL AND LIVER—CLINICAL RESEARCHO2.1RAPID DECLINE OF LIVER STIFFNESS WITH ALCOHOL WITHDRAWAL IN HEAVY DRINKERS

Background and aims. Measurement of liver stiffness using real-time elastography appears as a promising tool to evaluate the severity of chronic liver diseases. Previous studies in patients with alcoholic liver disease have suggested that fibrosis was the only histological parameter to influence liver stiffness. To challenge this hypothesis, we have prospectively tested the short-term impact of alcohol withdrawal on liver stiffness value. Methods. All patients hospitalized for alcohol withdrawal in our Liver Unit between September 2008 and December 2010 had a liver stiffness determination (using a FibroScan® device) at entry (D0) and 7 days after alcohol withdrawal (D7). Stiffness values were compared using non-parametric test for paired-values. We compared (i) the 10 measures performed at D0 and at D7 for each patient; (ii) the variation of the median result of all patients (using Wilcoxon test in both cases). Results. A total of 138 patients were included in the study [median alcohol consumption: 150g/day (range: 40-400); hepatitis C: n=22 (15.9%); cirrhosis: n=29 (21.0%)]. From D0 to D7, the liver stiffness decreased significantly in 61 patients (44.2%) and increased significantly in 18 (13.0%). Considering all patients, median liver stiffness value decreased from 7.25 to kPa (P<0.001). The stage of fibrosis indicated by liver stiffness changed in 47 patients between D0 and D7 (decrease in 33 and increase in 14). Conclusion. Liver stiffness decreases significantly in nearly half of alcoholic patients after only 7 days of abstinence. This result strongly suggests that non-fibrotic lesions (such as inflammatory ones) may influence liver stiffness. From a practical point of view, it also shows that variation in alcohol consumption must be taken into account for the interpretation of liver stiffness valu

Is cohorting the only solution to control carbapenemase-producing Enterobacteriaceae outbreaks? A single-centre experience

International audienceBACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) are a major health issue. Cohorting may help to control spread of CPEs in hospitals, but is expensive and hard to implement.AIM: To identify ward variables associated with CPE in-hospital transmission in a hospital where cohorting has never been implemented.METHODS: Cohort prospective study, comparing 14-consecutive-day periods regarding in-hospital transmission. Each period met the two following conditions: (i) CPE carriers/infected admitted for ≥48&nbsp;h; (ii) 80% of relative contact patients were screened at least twice. Periods (a) with no acquired CPE case among relative contact patients were compared to periods (b) during which one or more CPE case acquisition was identified. Variables potentially associated with CPE transmission were assessed: colonization pressure, caregiver:patient ratio, hand hygiene compliance, hydro-alcoholic product consumption, antibiotic consumption, and infection control team (ICT) involvement on the ward.FINDINGS: Sixty-eight periods of two consecutive weeks were included, 18 (26.5%) included at least one CPE case acquisition. By multivariate analysis, colonization pressure (odds ratio: 1.12; 95% confidence interval: 1.0-1.25; P&nbsp;=&nbsp;0.042) and antibiotic consumption (2.41; 1.02-5.66; P&nbsp;=&nbsp;0.044) were associated with CPE in-hospital transmission. Caregiver:patient ratio potentiated both these variables, suggesting a role for understaffing in CPE transmission.CONCLUSION: Understanding ward variables associated with CPE spread can help design suitable solutions. Colonization pressure and antibiotic consumption seems to be driving in-hospital transmission, along with caregiver:patient ratio. In presence of high colonization pressure, dedicated healthcare workers for managing CPE patients should be implemented. Co-ordination between ICT and antimicrobial stewardship team is also crucial to prevent CPE spread.</p

CHRNA5 as negative regulator of nicotine signaling in normal and cancer bronchial cells: Effects on motility, migration and p63 expression

Genome-wide association studies have linked lung cancer risk with a region of chromosome 15q25.1 containing CHRNA3, CHRNA5 and CHRNB4 encoding a3, a5 and ß4 subunits of nicotinic acetylcholine receptors (nAChR), respectively. One of the strongest associations was observed for a non-silent singlenucleotide polymorphism at codon 398 in CHRNA5. Here, we have used pharmacological (antagonists) or genetic (RNA interference) interventions to modulate the activity of CHRNA5 in nontransformed bronchial cells and in lung cancer cell lines. In both cell types, silencing CHRNA5 or inhibiting receptors containing nAChR a5 with a-conotoxin MII exerted a nicotine-like effect, with increased motility and invasiveness in vitro and increasing calcium influx. The effects on motility were enhanced by addition of nicotine but blocked by inhibiting CHRNA7, which encodes the homopentameric receptor a7 subunit. Silencing CHRNA5 also decreased the expression of cell adhesion molecules P120 and ZO-1 in lung cancer cells as well as the expression of DeltaNp63a in squamous cell carcinoma cell lines. These results demonstrate a role for CHRNA5 in modulating adhesion and motility in bronchial cells, as well as in regulating p63, a potential oncogene in squamous cell carcinoma. © The Author 2011. Published by Oxford University Press. All rights reserved

Azacitidine frontline therapy for unfit acute myeloid leukemia patients: Clinical use and outcome prediction

Hypomethylating agents are able to prolong the overall survival of some patients diagnosed with acute myeloid leukemia. The aim of this study was to evaluate the clinical use of azacitidine as front-line therapy in unfit acute myeloid leukemia (AML) patients and to develop a clinical prediction model to identify which patients may benefit more from the drug. One hundred and ten untreated unfit AML patients received front-line azacitidine therapy in Spain, and response and survival were evaluated in them following European LeukemiaNet (ELN) guidelines. A clinical prediction rule was obtained from this population that was validated and refined in 261 patients treated in France, Austria and Italy. ELN response was achieved in 21.0% of the 371 patients (CI95% 17.0-25.5) and did not depend on bone marrow blast cell percentage. Median overall survival was 9.6 months (CI95% 8.5-10.8) and 40.6% of the patients were alive at 1 year (CI95% 35.5-45.7). European ALMA score (E-ALMA), based on performance status, white blood cell counts at azacitidine onset and cytogenetics, discriminated three risk groups with different survival and response rates. Azacitidine seems a reasonable therapeutic option for most unfit AML patients, i.e. those displaying a favorable or intermediate E-ALMA score

Projet ClimAware : impacts du changement climatique sur la gestion des ressources en eau. Stratégies régionales et vision européenne. Rapport final

The main objectives of the project have been addressed by combining a European modelling approach with case study analysis and regional (local) knowledge of water demand and water availability considering climate change as well as socio-economic developments. An integrated assessment for entire Europe was performed under the consideration of different scenarios and cli-mate change projections. This large-scale perspective allows indicating regions which are po¬ten-tially vulnerable to climate change and furthermore to identify regional adaptation measures which could be promoted at the EU level. Additionally three case studies were selected across Europe to in¬vestigate changes in hydrologic regimes, water availability and sectoral water use. These case stu¬dies are focusing on three different water management issues in three different regions. In the first case study, the influence of climate change on the hydromorphological conditions according to the WFD were evaluated for a section of the Eder River (Germany). The objective of this case study was to examine whether the environmental WFD objectives can be achieved in a typical river section considering climate change impacts. The second case study investigated water management, especially drinking water provision, and flood alleviation in the Seine river basin (France), which is partly based on the operation of artificial reservoirs. Scenarios were developed linking the impact of climate change on water resources and changes in water demand and its management. The third case study assesses the quantitative effects of climate change on water balance components and water use in the agricultural sector of the Italian Apulia region, in order to support the adoption of adaptation measures. Actually, in the Apulia region agriculture still remains the primary user of water and the primary economical resource.Les principaux objectifs du projet ont été adressés en combinant une approche de modélisation européenne avec une analyse de cas d'études et la connaissance régionale (locale) des demandes et disponibilités en eau en considérant le changement climatique ainsi que les développements socio-économiques. Une estimation intégrée pour l'Europe entière a été entreprise en considérant différents scénarios et projections de changement climatique. Cette perspective à large échelle permet d'identifier des régions qui sont potentiellement vulnérables au changement climatique mais aussi d'identifier les mesures d'adaptation qui pourraient être développées au niveau européen. De plus, trois cas d'étude ont été sélectionnés à travers l'Europe pour étudier les changements dans les régimes hydrologiques, la disponibilité en eau, et les usages de l'eau. Ces cas d'étude concernent trois différents problèmes de gestion de l'eau dans trois régions différentes. Dans le premier cas d'étude, l'influence du changement climatique sur les conditions hydromorphologiques selon la DCE ont été évaluées pour une section de la rivière Eder (Allemagne). L'objectif de ce cas d'étude était d'examiner si l'objectif environnemental de la DCE peut être rempli dans une section typique de la rivière en considérant le changement climatique. Le second cas d'étude s'est penché sur la gestion de l'eau, plus spécialement de la fourniture en eau potable, et sur la mitigation des crues sur le bassin de la Seine (France), qui est partiellement basée sur l'exploitation de réservoirs artificiels. Des scénarios ont été développés pour évaluer l'impact du changement climatique sur les ressources en eau en testant différents modes de gestion des réservoirs. Le troisième cas d'étude a évalué les effets quantitatifs du changement climatique sur les composantes du bilan en eau et sur l'utilisation de l'eau sur le secteur agricole de la région des Pouilles en Italie, afin de soutenir l'adoption de mesures d'adaptation. En fait, la région des Pouilles l'agriculture reste le principal utilisateur d'eau et la principale ressource économique