3' RNA ligase mediated rapid amplification of cDNA ends for validating viroid induced cleavage at the 3' extremity of the host mRNA

Abstract : 5′ RNA ligase-mediated rapid amplification of cDNA ends (5′ RLM-RACE) is a widely-accepted method for the validation of direct cleavage of a target gene by a microRNA (miRNA) and viroid-derived small RNA (vd-sRNA). However, thi\s method cannot be used if cleavage takes place in the 3′ extremity of the target RNA, as this gives insufficient sequence length to design nested PCR primers for 5′ RLM RACE. To overcome this hurdle, we have developed 3′ RNA ligase-mediated rapid amplification of cDNA ends (3′ RLM RACE). In this method, an oligonucleotide adapter having 5′ adenylated and 3′ blocked is ligated to the 3′ end of the cleaved RNA followed by PCR amplification using gene specific primers. In other words, in 3′ RLM RACE, 3′ end is mapped using 5′ fragment instead of small 3′ fragment. The method developed here was verified by examining the bioinformatics predicted and parallel analysis of RNA ends (PARE) proved cleavage sites of chloride channel protein CLC-b-like mRNA in Potato spindle tuber viroid infected tomato plants. The 3′ RLM RACE developed in this study has the potential to validate the miRNA and vd-sRNA mediated cleavage of mRNAs at its 3′ untranslated region (3′ UTR)

Évolution clinico-biologique de 73 femmes diagnostiquées infectées par le VIH pendant la grossesse et traitées par trithérapie antirétrovirale (étude au Congo-Brazzaville)

La pandémie VIH est un problème de santé publique majeur en Afrique subsaharienne. Les programmes de Prévention de la Transmission de la Mère à l Enfant consistent à instaurer un traitement antirétroviral pendant la grossesse pour prévenir la transmission verticale du virus. Peu de données sont disponibles sur l évolution de ces patientes après l'accouchement. Cette étude rétrospective et descriptive a été menée au Centre de Traitement Ambulatoire de la Croix-Rouge Française situé à Pointe-Noire en République du Congo. Elle s est intéressée au suivi de 73 femmes traitées soit à visée prophylactique avec arrêt du traitement à l accouchement ou à la fin de l allaitement maternel (30 patientes), soit mises sous TAR au long cours pour leur propre santé (43 patientes). Elles avaient un niveau d éducation bas, la plupart étaient femmes au foyer/ménagères (44%), ou menaient un petit commerce (27,4%). Le suivi clinique a été bon avec 100% de patientes vues à M24, 84% à M30 et 81% à M36. Le suivi biologique (CD4 et CV) a été irrégulier en raison des problèmes d approvisionnement en réactifs, malgré les capacités techniques et humaines disponibles. Trois décès ont été relevés sur 36 mois. L évolution clinique des femmes ayant arrêté le TAR ne semblait pas péjorée par rapport à celle des femmes qui l ont poursuivi. Aucune des patientes traitées en prophylaxie n a présenté d effet secondaire au traitement, versus 6 événements pour les patientes traitées au long cours. La réflexion en vue d améliorer les conduites à tenir thérapeutiques nécessite d être élargie en prenant en compte, l ensemble des critères économiques, sociaux et culturels distinctifs de l Afrique subsaharienne.The HIV pandemic is a main public health concern in sub-Saharan Africa. The Prevention of Mother to Child Transmission programs aim to treat pregnant women with AntiRetroviral Treatment, in order to avoid the vertical virus transmission. Few data are available about the clinical and biological follow-up after delivery. This retrospective and descriptive study was conducted in the day care centre run by the French Red Cross in Pointe-Noire, Republic of Congo. Seventy-three patients were included. They were given either a prophylactic treatment which was stopped after delivery or breastfeeding (30 patients), or a long-term treatment for their own health (43 patients). The education level was low; most of them were housewives or housecleaners (44%), or had a street seller activity (27,4%). The clinical follow-up was good as 100 % of patients went to their consultation on M24, 84% on M30 and 81% on M36. The biological follow-up based on CD4 and Viral Load measure was rarely performed, due to the supply chain failures of biologic reactive, and despite the good technic and human support available. Three deaths occurred during the 36 months of follow-up. The clinical evolution of patients who stopped the ARV seems to be as safe as evolution of women who kept on with treatment. No patient treated with prophylactic ARV went through adverse effect, compared to 6 women in the life-time treatment group. In order to improve the guidelines, we need to take into account, further than the biomedical part, the economic, social and cultural aspects of sub-Saharan African context.GRENOBLE1-BU Médecine pharm. (385162101) / SudocSudocFranceF

Louis Bachelier On the centenary of Théorie de la Spéculation

International audienceWritten on the occasion of the centenary of Louis Bachelier's 1900 PhD thesis “Théorie de la spéculation”, this paper puts Bachelier into a historical perspective. It explains his role as a pioneer in both mathematical finance and probability theory, and it also gives a careful account of Bachelier's difficult personal and scientific career. This includes a discussion of Poincaré's report on the PhD thesis (the report itself with an English translation is reproduced in the appendix), and an explanation for the controversy between Bachelier's work and Paul L´evy's reports on it. The paper also contains a curriculum vitae and a list of the publications of Bachelier is made available

Centre d’anthropologie

Jean-Pierre Albert, directeur d’études avec Dominique Blanc, ingénieur d’études Approches anthropologiques du travail social Le séminaire a été construit, en alternance, autour, d’une part, d’apports théoriques et de débats concernant la pertinence d’un regard anthropologique et d’une recherche de type ethnographique dans l’analyse des « questions sociales » et des « interventions sur le social » et, d’autre part, de l’exposé de recherches en cours, telles que, cette année : les processus soc..

Retrospective French nationwide survey of childhood aggressive vascular anomalies of bone, 1988-2009

<p>Abstract</p> <p>Objective</p> <p>To document the epidemiological, clinical, histological and radiological characteristics of aggressive vascular abnormalities of bone in children.</p> <p>Study design</p> <p>Correspondents of the French Society of Childhood Malignancies were asked to notify all cases of aggressive vascular abnormalities of bone diagnosed between January 1988 and September 2009.</p> <p>Results</p> <p>21 cases were identified; 62% of the patients were boys. No familial cases were observed, and the disease appeared to be sporadic. Mean age at diagnosis was 8.0 years [0.8-16.9 years]. Median follow-up was 3 years [0.3-17 years]. The main presenting signs were bone fracture (n = 4) and respiratory distress (n = 7), but more indolent onset was observed in 8 cases. Lung involvement, with lymphangiectasies and pleural effusion, was the most frequent form of extraosseous involvement (10/21). Bisphosphonates, alpha interferon and radiotherapy were used as potentially curative treatments. High-dose radiotherapy appeared to be effective on pleural effusion but caused major late sequelae, whereas antiangiogenic drugs like alpha interferon and zoledrenate have had a limited impact on the course of pulmonary complications. The impact of bisphosphonates and alpha interferon on bone lesions was also difficult to assess, owing to insufficient follow-up in most cases, but it was occasionally positive. Six deaths were observed and the overall 10-year mortality rate was about 30%. The prognosis depended mainly on pulmonary and spinal complications.</p> <p>Conclusion</p> <p>Aggressive vascular abnormalities of bone are extremely rare in childhood but are lifethreatening. The impact of anti-angiogenic drugs on pulmonary complications seems to be limited, but they may improve bone lesions.</p

Étude critique des tests de résistance génotype chez 12 patients VIH en échec virologique au traitement antirétroviral

GRENOBLE1-BU Médecine pharm. (385162101) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

Durée de traitement des érysipèles (facteur de risque de récidive à 3 mois)

GRENOBLE1-BU Médecine pharm. (385162101) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

L'utilisation des antibiotiques en dermatologie (étude auprès des dermatologues des départements Isère, Savoie et Haute-Savoie)

La sélection et l émergence de bactéries multirésistantes sont directement liées à la consommation d antibiotiques. De cette constatation, nous avons voulu savoir si nous utilisions correctement les antibiotiques dans la pathologie infectieuse cutanée communautaire. Pour répondre à cette question nous avons envoyé un questionnaire aux dermatologues des départements d Isère, de Savoie et de Haute-Savoie, concernant leur thérapeutique ou leur conduite tenue dans l érysipèle, l impétigo, les folliculites et furoncles, les plaies aiguës et chroniques. Il nous a semblé également important de discuter de l antibiothérapie dans l acné. Nous avons comparé nos résultats aux données de la littérature. De cette analyse, il ressort que nous ne suivons pas les conférences de consensus ni les recommandations élaborées par la communauté scientifique. Cette étude soulève donc le problème de l accès à cette information et de notre capacité à modifier nos habitudes de prescription.GRENOBLE1-BU Médecine pharm. (385162101) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF

Carbapenem use in French hospitals: A nationwide survey at the patient level.

International audienceThe objective of this study was to evaluate the characteristics of carbapenem use in French healthcare settings in order to guide future actions. Healthcare facilities voluntarily participated in a nationwide cross-sectional survey in 2011. Medical data and reasons for carbapenem treatment (CPR) and discontinuation were recorded for all patients treated with carbapenems. A total of 2338 patients were recorded by 207 facilities. The median duration of CPR was 8 days, and 31.4% of patients received CPR for >10 days. An antibiotic consultant was involved in the initial choice of CPR in 36.8% of cases. CPR was chosen on an empirical (EP) basis for 1229 patients (52.6%), mainly because of severe sepsis (48.6%) or a perceived risk of bacterial resistance (33.7%). Among EP patients, de-escalation was more frequent in the case of intervention of an antibiotic consultant (35.1%) than without intervention (22.9%) (P<0.01). Among the 1109 patients receiving CPR initially based on bacteriological results, 607 (54.7%) had ESBL-producing Enterobacteriaceae and 397 (35.8%) had Gram-negative bacilli susceptible to at least one β-lactam other than carbapenems or to fluoroquinolones. Among the latter, de-escalation was performed in 59 cases (14.9%). The intervention of an antibiotic consultant did not favour de-escalation in this group. In conclusion, carbapenems are frequently used for treating suspected or confirmed multidrug-resistant bacteria, and overall CPR duration is long. De-escalation is frequently not implemented despite isolates being susceptible to other drugs. More frequent antibiotic consultant intervention may help to decrease carbapenem use in the case of EP treatment

Insights Into Potato Spindle Tuber Viroid Quasi-Species From Infection to Disease

Viroids are non-coding RNA plant pathogens that are characterized by their possession of a high mutation level. Although the sequence heterogeneity in viroid infected plants is well understood, shifts in viroid population dynamics due to mutations over the course of infection remain poorly understood. In this study, the ten most abundant sequence variants of potato spindle tuber viroid RG1 (PSTVd) expressed at different time intervals in PSTVd infected tomato plants were identified by high-throughput sequencing. The sequence variants, forming a quasi-species, were subjected to both the identification of the regions favoring mutations and the effect of the mutations on viroid secondary structure and viroid derived small RNAs (vd-sRNA). At week 1 of PSTVd infection, 25% of the sequence variants were similar to the "master" sequence (i.e., the sequence used for inoculation). The frequency of the master sequence within the population increased to 70% at week 2 after PSTVd infection, and then stabilized for the rest of the disease cycle (i.e., weeks 3 and 4). While some sequence variants were abundant at week 1 after PSTVd infection, they tended to decrease in frequency over time. For example, the variants with insertions at positions 253 or 254, positions that could affect the Loop E as well as the metastable hairpin I structure that has been shown important during replication and viroid infectivity, resulted in decreased frequency. Data obtained byin silicoanalysis of the viroid derived small RNAs (vd-sRNA) was also analyzed. A few mutants had the potential of positively affecting the viroid's accumulation by inducing the RNA silencing of the host's defense related genes. Variants with mutations that could negatively affect viroid abundance were also identified because their derived vd-sRNA were no longer capable of targeting any host mRNA or of changing its target sequence from a host defense gene to some other non-important host gene. Together, these findings open avenues into understanding the biological role of sequence variants, this viroid's interaction with host components, stable and metastable structures generated by mutants during the course of infection, and the influence of sequence variants on stabilizing viroid population dynamics