First report of eprinomectin-resistant isolates of Haemonchus contortus in 5 dairy sheep farms from the Pyrénées Atlantiques département in France.

Infection of sheep by gastrointestinal nematodes (GIN) in pastoral systems such as those found in the South Western area of France, the Pyrénées Atlantiques, is one of the main reasons for economic loss and degradation of their welfare. In the present study, the efficacy of eprinomectin (EPN) was monitored on farms from this area following suspicion of lack of anthelmintic efficacy. Suspicions were raised by veterinarians, based on clinical signs ranging from milk and body condition loss, to anaemia, and mortality. Resistance was evaluated according to the World Association for the Advancement for Veterinary Parasitology (WAAVP) guidelines using fecal egg count reduction tests reinforced by individual analysis of drug concentration in the serum of all treated ewes by high-performance liquid chromatography (HPLC). EPN was administered by subcutaneous (SC) and topical (T) route according to manufacturer's requirements, as well as by the oral route (O) with the topical solution according to off-labelled practices in the field. For the first time in France, the presence of resistant isolates of Haemonchus contortus to EPN was observed in 5 dairy sheep farms. The HPLC dosages showed exposure of worms to concentrations compatible with anthelmintic activity for animals treated by the SC and O routes. By contrast, they showed under exposure to the drug of most individuals treated by the T route. EPN is the only null milk withdrawal anthelmintic molecule currently available. The presence of resistant isolates of the pathogenic H. contortus to EPN in this important dairy region requires an urgent change in grazing, and sometimes production, systems.

Inhibition of OCT2, MATE1 and MATE2-K as a possible mechanism of drug interaction between pazopanib and cisplatin.

We hypothesized that pazopanib is an inhibitor of cisplatin renal transporters OCT2, MATE1 and MATE2-K based on previous studies demonstrating an interaction between tyrosine kinase inhibitors and these transporters. Because several combinations of targeted therapies and cytotoxics are currently in development for cancer treatment, such an interaction is worth investigating. Experiments on HEK293 cells stably transfected to express OCT2, MATE1, MATE2-K or an empty vector (EV) were conducted. The inhibitory effect of pazopanib on these transporters was measured using the uptake of fluorescent substrate ASP+ and cisplatin in the different cell lines. The effect of pazopanib on cisplatin-induced cytotoxicity was also evaluated. A decrease of ASP+ uptake was observed in OCT2-HEK, MATE1-HEK and MATE2K-HEK cell lines after addition of pazopanib at increasing concentrations. Pazopanib inhibited cisplatin specific uptake in OCT2-HEK, MATE1-HEK and MATE2K-HEK lines. Cytotoxicity experiments showed that co-incubation of cisplatin with pazopanib multiplied up to 2.7, 2.4 and 1.6 times the EC50 values of cisplatin in OCT2-HEK, MATE1-HEK and MATE2K-HEK cell lines respectively, reaching about the same values as in EV-HEK cells. To conclude, pazopanib inhibits OCT2, MATE1 and MATE2-K, which are involved in cisplatin secretion into urine. The combination of these two drugs may lead to an interaction and increase the cisplatin-induced systemic toxicity. Given the wide variability of plasma pazopanib concentrations observed in vivo, the interaction may occur in a clinical setting, particularly in overexposed patients. The existence of a drug-drug interaction should be investigated when pazopanib is associated with a substrate of these transporters.

[Evolution of food supply (apart from school catering) between 2004/2005 and 2009/2010 in middle- and high-schools of Aquitaine, France]. / Évolution de l'offre alimentaire hors restauration scolaire entre 2004-2005 et 2009-2010 dans les établissements du second degré d'Aquitaine.

BACKGROUND: The aim of this study was to assess the evolution of food supply (apart from school catering) between school years 2004/2005 and 2009/2010, in middle- and high-schools from the Aquitaine region (southwest France), in order to evaluate the impact of actions conducted within the framework and the program Nutrition, Prevention and Health of children and adolescents in Aquitaine (southwest France). METHODS: Two surveys were carried out among all middle- and high-schools of the Aquitaine region in 2004/2005 (n=536) and 2009/2010 (n=539) within the framework of a regional multidisciplinary public health program "Nutrition, prevention and health of children and teenagers in Aquitaine". For both 2004/2005 and 2009/2010, data were collected using the same questionnaire and dealt with school characteristics and modalities of food supply (apart from school catering). RESULTS: Response rate was 84.1% in 2004/2005 and 79.6% in 2009/2010. The proportion of schools offering food to pupils (apart from school catering) significantly decreased in 5 years (from 80.1% to 50.1%, P<0.001). Between 2004/2005 and 2009/2010, we observed a stabilization in the proportion of schools offering free food (from 19.7 to 17%, P=0.3), a significant decrease of those selling food (from 62.8 to 37.1, P<0.001), offering vending machines (from 43.5 to 7.2, P<0.001) and a significant increase of those offering water supply (from 8.2 to 44%, P<0.001). The composition of each modality of food supply (apart from school catering) has also been improved: less sweet and fat food, more bread and fruits. CONCLUSION: This study shows an overall improvement of food supply apart from school catering (food sale, free food and vending machines) in middle- and high-schools from the Aquitaine region (southwest France) between 2004/2005 and 2009/2010. This improvement is related to the proportion of schools offering food (quantitative improvement), as well as to the composition of food supply (qualitative improvement). These results show an improvement of food supply (apart from school catering), suggesting that actions implemented in the framework of the program "Nutrition, prevention and health of children and adolescents in Aquitaine" may have led to these improvements.

Electrochemical investigation of the P2­NaxCoO2 phase diagram.

Sodium layered oxides NaxCoO2 form one of the most fascinating low-dimensional and strongly correlated systems; in particular P2­NaxCoO2 exhibits various single-phase domains with different Na+/vacancy patterns depending on the sodium concentration. Here we used sodium batteries to clearly depict the P2­NaxCoO2 phase diagram for x≥0.50. By coupling the electrochemical process with an in situ X-ray diffraction experiment, we identified the succession of single-phase or two-phase domains appearing on sodium intercalation with a rather good accuracy compared with previous studies. We reported new single-phase domains and we underlined the thermal instability of some ordered phases from an electrochemical study at various temperatures. As each phase is characterized by the position of its Fermi level versus the Na+/Na couple, we showed that the synthesis of each material, even in large amounts, can be carried out electrochemically. The physical properties of the as-prepared Na1/2CoO2 and Na2/3CoO2 ordered phases were characterized and compared. Electrochemical processes are confirmed to be an accurate route to precisely investigate in a continuous way such a complex system and provide a new way to synthesize materials with a very narrow existence range.

Epidemiology and evolution of antibiotic resistance of Haemophilus influenzae in children 5 years of age or less in France, 2001-2008: a retrospective database analysis.

Trends in the evolution of antimicrobial resistance and mechanisms of resistance of Haemophilus influenzae to ß-lactam antibiotics in France were assessed through a retrospective database review. The antimicrobial resistance of 2,206 H. influenzae strains from children aged ≤5 years was studied between 2001 and 2008. Strains were isolated from blood or cerebrospinal fluid (n = 170), bronchial secretions (n = 188), middle ear fluid, and nasopharynx or conjunctiva (n = 1,848). A proportion of 95.1 % (n = 2,097) were non-typeable H. influenzae (NTHi). ß-lactamase production was identified in 27.5 % of NTHi isolates (all TEM-1), while ß-lactamase-negative ampicillin resistance and ß-lactamase-negative amoxicillin-clavulanate resistance among NTHi was 16.9 and 6.4 %, respectively. Over time, a statistically significant decrease in ß-lactamase-producing strain prevalence (p < 0.0001) and a statistically significant increase in ß-lactamase-negative ampicillin-resistant (BLNAR) strains (p < 0.0001) were observed in NTHi isolates from 2001 to 2008. The largest changes coincided with a campaign to reduce antibiotic use in France. An increasing diversity of amino acid substitution patterns was observed, with the emergence of group III/'III-like' patterns linked to high-level resistance. In France, amino acid substitution patterns are increasingly diverse, and strains with high-level antibiotic resistance are emerging. This study highlights the complexity of resistance dynamics within a given country. These results have implications on antibiotic guidelines and illustrate the importance of continued surveillance.

Reactivity of (1→3)-ß-d-glucan assay in bacterial bloodstream infections.

To diagnose invasive fungal infections, the detection of (1 → 3)-ß-d-glucan in serum has shown variable specificity, depending on the targeted population. Several circumstances for false-positive results of beta-glucan tests have been identified, among which are severe bacterial infections. In this study, we measured (1 → 3)-ß-d-glucan by the Fungitell test in the serum of 62 patients (one serum sample tested per patient) for whom invasive fungal infection was not suspected: 19 control subjects and 43 patients with bacteraemia. The test was interpretable for 58 sera: all 19 control subjects had negative beta-glucan test; among the 39 bacteraemic patients, we report 16 false-positive results. For the 22 patients undergoing bacteraemia due to Gram-negative bacilli, we observed 13 false-positive results (59%). Among the 17 patients with bloodstream infection involving Gram-positive cocci, three false-positive tests were recorded, but none in the eight cases of Streptococcus pneumoniae bacteraemia. Statistical analysis showed that beta-glucan levels were significantly higher in patients with Gram-negative bacilli bloodstream infection in comparison to those with bacteraemia due to Gram-positive cocci. These results were independent from other previously described causes for false-positive beta-glucan tests. These data might help physicians to interpret positive beta-glucan detection when an invasive fungal infection is suspected, especially for patients with bacterial infections.

Reinvestigation of the OP4-(Li/Na)CoO2-layered system and first evidence of the (Li/Na/Na)CoO2 phase with OPP9 oxygen stacking.

The composition and synthesis conditions of the (Li/Na)CoO(2) phase with an ordered 1:1 Li/Na stacking alternating with CoO(2) slabs were determined from a careful study of the P2-Na(∼0.7)CoO(2)-O3-LiCoO(2) system. An in situ X-ray diffraction (XRD) thermal study emphasizes the metastable character of this phase that can be stabilized only by very fast quenching. Its composition, (Li(0.42)Na(0.37))CoO(2), is significantly different from the ideally expected one, (Li(0.50)Na(0.35))CoO(2), and its structure, confirmed by Rietveld refinement of the XRD pattern, presents an ideal alternate ordering of lithium, cobalt, and sodium layers within OP4-type oxygen packing. The presence of vacancies in both alkali-ion layers was confirmed by electrochemical intercalation of lithium and sodium. For the first time, a new type of layered oxide exhibiting OPP9-type oxygen packing was evidenced. Between the CoO(2) slabs, alkali ions are intercalated in the following order: Li(octa)-Na(prism)-Na(prism). This material crystallizes in the R3m space group with a(hex) = 2.828 Šand c(hex) = 46.85 Šcell parameters.

First experimental evidence of a new D4-AgCoO2 delafossite stacking.

A new polytype of AgCoO(2) delafossite has been prepared from the ordered OP4-(Li/Na)CoO(2) layered compound using ion-exchange reaction in molten salts. As expected from the structural model assuming a topotactic process, the lamellar structure of this new polytype is an alternate combination of the already known 2H and 3R delafossite polytypes. It crystallizes in the P6(3)/mmc space group with cell parameters a(hex.) = 2.871(1) Å and c(hex.) = 24.448(1) Å. Thermal stability, morphology characterization, and electrical properties are reported here and compared with those of the 2H and 3R AgCoO(2) polytypes.

(Li/Ag)CoO2: a new intergrowth cobalt oxide composed of rock salt and delafossite layers.

A new ordered (Li/Ag)CoO(2) layered compound with an unusual oxygen packing combining rock salt and delafossite layers is obtained during the (Li(+), Na(+))/Ag(+) ionic exchange from the OP4-(Li/Na)CoO(2) precursor. This compound is actually an intermediate step to the final D4-AgCoO(2) delafossite and can be isolated thanks to the kinetics difference between the Li(+)/Ag(+) and Na(+)/Ag(+) exchange processes. It crystallizes in the P6(3)/mmc space group with cell parameters a(hex.) = 2.848(3) Å and c(hex.) = 21.607(7) Å. The details of the structure as well as its thermal stability and transport properties are presented and discussed.

Congenital toxoplasmosis in France in 2007: first results from a national surveillance system.

When immunocompetent people become infected with the parasite Toxoplasma gondii, the disease is generally asymptomatic. However, transplacental transmission of T. gondii may lead to severe congenital infection including in utero abortion, foetal death, or neurological or ocular damage of the foetus. France has had a national programme to prevent congenital toxoplasmosis since 1978. However, although estimated seroprevalence in pregnant women has fallen from 84% in the 1960s to 44% in 2003, no reliable data have been available on the annual number of cases of congenital toxoplasmosis or the severity of infection. In 2006, the French National Institute for Public Health Surveillance (Institut de Veille Sanitaire) and the National Reference Centre for Toxoplasmosis recommended that a national laboratory-based surveillance system be used for the surveillance of the disease. In 2007, 31 laboratories reported at least one congenital case through the surveillance system, giving a total of 272 cases. A total of 11 terminations of pregnancy were reported (six abortions and five foetal deaths). Of the live-born cases, 206 were asymptomatic, 28 were symptomatic and seven had a severe form of the disease. As there were 818,700 births in France and French overseas departments in 2007, the overall prevalence of congenital toxoplasmosis observed that year was 3.3 (95% confidence interval (CI): 2.9 to 3.7) per 10,000 live births and the incidence rate of the disease at birth was 2.9 (95% CI: 2.5 to 3.2) per 10,000 live births; the estimated incidence rate of symptomatic congenital toxoplasmosis was 0.34 (95% CI: 0.2 to 0.5) cases per 10,000 live births.

Lithium deintercalation in LiFePO4 nanoparticles via a domino-cascade model.

Lithium iron phosphate is one of the most promising positive-electrode materials for the next generation of lithium-ion batteries that will be used in electric and plug-in hybrid vehicles. Lithium deintercalation (intercalation) proceeds through a two-phase reaction between compositions very close to LiFePO(4) and FePO(4). As both endmember phases are very poor ionic and electronic conductors, it is difficult to understand the intercalation mechanism at the microscopic scale. Here, we report a characterization of electrochemically deintercalated nanomaterials by X-ray diffraction and electron microscopy that shows the coexistence of fully intercalated and fully deintercalated individual particles. This result indicates that the growth reaction is considerably faster than its nucleation. The reaction mechanism is described by a 'domino-cascade model' and is explained by the existence of structural constraints occurring just at the reaction interface: the minimization of the elastic energy enhances the deintercalation (intercalation) process that occurs as a wave moving through the entire crystal. This model opens new perspectives in the search for new electrode materials even with poor ionic and electronic conductivities.

Increased risk of colonic neoplasia in patients with sporadic duodenal adenoma.

BACKGROUND: Recent studies have shown an increased risk of colorectal neoplasia in patients with duodenal neoplasia. The aim of this retrospective case-control study was to confirm this risk. PATIENTS AND METHODS: Rate of colorectal neoplasia in 29 patients with one or more duodenal adenomas were compared with controls matched for gender and age, but without duodenal adenomas (one case to two controls). Patients with neoplasia of the ampulla, familial adenomatous polyposis or other known hereditary conditions of the digestive tract were excluded. Indications for upper and lower gastrointestinal endoscopy in controls were abdominal pain or changes in bowel habits. Controls with anemia or digestive bleeding were not included. Neoplastic lesions found at colonoscopy were classified as adenomas, advanced adenomas (size > or =10 mm, villous component, high-grade dysplasia), cancers and advanced neoplasia (cancers and advanced adenomas). Comparison between groups was by Fisher's exact test or Student's t test. Odds-ratios (OR) and 95% confidence intervals were calculated, if the difference was significant. RESULTS: Mean age of the 29 cases (seven women, 22 men) was 63.2 years and that of the 58 controls (14 women, 44 men) was 62.5 years. First-degree family history of colorectal cancer was present in four cases (13.8%) and eight controls (13.8%) (NS). Colonoscopy showed at least one adenoma in 15 cases (51.7%) and 11 controls (19%) (P=0.0027; OR 1.87, 1.0-3.49), advanced adenomas in four cases (13.8%) and three controls (5.2%) (NS), and colonic adenocarcinoma in three cases (10.3%) and no controls (0%) (P=0.03). Advanced neoplasia was present in seven cases (24.1%) and three controls (5.2%) (P=0.014; OR 2.86, 0.96-8.52). Results were not significantly modified after the exclusion of patients with a family history of colorectal cancer. CONCLUSION: Although lacking in statistical power, these results confirm that patients with sporadic duodenal adenoma are at high risk of colonic adenoma and advanced neoplasia, warranting systematic colonoscopy.

[Allergic respiratory diseases related to mould in the home]. / Pathologies allergiques respiratoires liées aux moisissures de l'habitat.

Exposure to mould is a potential risk factor for asthma in both adults and children. In adult, the relation between exposure, sensitization and symptoms has been demonstrated in severe asthmatics sensitized to Alternaria. For children, exposure to mould in childhood is a risk factor for asthma in both atopic and non-atopic individuals. Exposure or sensitization to moulds are a risk factor for severe asthma and/or exacerbations in children. There appears to be a causal relationship between exposure and asthma. This link seems less significant in adults. However, in adults mould sensitive asthma seems to determine a phenotype of severe asthma associated with more marked obstructive lung disease. Moulds can stimulate either innate or the acquired immunity. They are responsible for a marked Th2 inflammation leading to more severe asthma. Besides the immunological mechanisms, toxic mechanisms can also intervene. It is therefore not correct to reduce the effect of moulds, particularly in respiratory symptoms, to only allergic mechanisms.

[Vocal cord dyskinesia and/or asthma]. / Dyskinésie des cordes vocales et/ou asthme.

INTRODUCTION: Vocal cord dyskinesia or vocal cord dysfunction (VCD) is characterized by intermittent abnormal adduction of the vocal cords leading to airflow limitation at the level of the larynx, in the absence of local organic disease. It may occur in isolation or in association with asthma. The pathophysiology is complex and poorly understood. Wheeze, stridor or apparent upper airway obstruction are the most common symptoms. It occurs in a wide age range, more commonly in women, and diagnosis is often delayed and leads to unnecessary treatments (intubation, tracheostomy and high dose steroids). METHODS: A retrospective study of 15 cases of VCD (8 cases of isolated VCD and 7 cases of VCD with associated asthma) describing the main clinical features and the diagnosis strategy. RESULTS: Apparent upper airway obstruction, with or without associated asthma, requires an ear nose and throat examination with laryngoscopy to confirm the paradoxical adduction of the vocal cords during an acute episode of dyspnoea or during a provocation test with triggers like exercise or exposure to irritants, and for the purpose of differential diagnosis. CONCLUSIONS: VCD remains under-appreciated and misdiagnosed, often by mimicking asthma with which it can be associated. A delayed diagnosis by emergency specialists, pulmonologists and ear nose and throat surgeons leads to unnecessary treatments and morbidity before specific therapy can be given.

3-month prognostic impact of severe acute renal failure under veno-venous ECMO support: Importance of time of onset.

PURPOSE: Veno-venous ECMO is increasingly used for the management of refractory ARDS. In this context, acute kidney injury (AKI) is a major and frequent complication, often associated with poor outcome. We aimed to identify characteristics associated with severe renal failure (Kidney Disease Improving Global Outcome (KDIGO) 3) and its impact on 3-month outcome. METHODS: Between May 2009 and April 2016, 60 adult patients requiring VV-ECMO in our University Hospital were prospectively included. RESULTS: AKI occurrence was frequent (75%; n=45), 51% of patients (n=31) developed KDIGO 3 - predominantly prior to ECMO insertion - and renal replacement therapy was required in 43% (n=26) of cases. KDIGO 3 was associated with a lower mechanical ventilation weaning rate (24% vs 68% for patients with no AKI or other stages of AKI; p<0.001) and a higher 90-day mortality rate (72% vs 32%, p=0.002). Multivariate logistic regression suggested that KDIGO 3 occurrence prior to ECMO insertion, as well as PaCO2>57mmHg and mSOFA>12 were independent risks factors for 90-day mortality. CONCLUSION: KDIGO 3 AKI occurrence is correlated with the severity of patients' clinical condition prior to ECMO insertion and is negatively associated with 90-day survival.

[Epidemiology of Haemophilus influenzae strains collected in 2004 in France and in vitro assessment of their susceptibility to antibiotics]. / Epidémiologie et évaluation de la sensibilité aux antibiotiques de souches d'Haemophilus influenzae isolées en 2004 en France.

OBJECTIVES: The aim of this study was to describe the epidemiology of H. influenzae strains collected in 2004 at the National Reference Center and to evaluate their susceptibility to various antibiotics. METHODS: Demographic and clinical characteristics, capsular serotyping by slide agglutination with specific antisera, beta-lactamase by a chromogenic cephalosporin test (Nitrocefin) and MICs of amoxicillin, co-amoxiclav, cefpodoxime, cefaclor, cefuroxime, cefotaxime, erythromycin, pristinamycin and telithromycin by agar dilution method on Haemophilus Test Medium were determined for each strain. RESULTS: 807 strains of H. influenzae were identified: 41.8% from bronchial secretions (BS), 16.2% from conjunctivitis, 6.6% from otitis media (OM), 4.2% from CSF and 8.6% from blood cultures. 95.6% of strains was not capsulated and 4.4% was of serotype b, e, or f. 26.3% of strains was beta-lactamase producing (TEM type). 185 isolates (22.8% of total strains) had reduced susceptibility to beta-lactams due to modification of the target associated or not with beta-lactamase production. When beta-lactamase was produced, the MICs of amoxicillin increased, but the activity of the other antibiotics was unchanged. Low BLNAR strains showed an increase in the MICs of all beta-lactams. This increase was weak and variable according to beta-lactams. Pristinamycin and telithromycin activities were unchanged against these strains. Two strains were resistant to erythromycin. CONCLUSIONS: Theses results show that both beta-lactamase and modifications of the target are widespread among H. influenzae strains isolated in France. Cefpodoxime remains the most active compounds against H. influenzae, whatever the resistance mechanisms, followed by pristinamycin, telithromycin, and co-amoxiclav.

Farnesylated RhoB inhibits radiation-induced mitotic cell death and controls radiation-induced centrosome overduplication.

Our previous results demonstrated that expressing the GTPase ras homolog gene family, member B (RhoB) in radiosensitive NIH3T3 cells increases their survival following 2 Gy irradiation (SF2). We have first demonstrated here that RhoB expression inhibits radiation-induced mitotic cell death. RhoB is present in both a farnesylated and a geranylgeranylated form in vivo. By expressing RhoB mutants encoding for farnesylated (RhoB-F cells), geranylgeranylated or the CAAX deleted form of RhoB, we have then shown that only RhoB-F expression was able to increase the SF2 value by reducing the sensitivity of these cells to radiation-induced mitotic cell death. Moreover, RhoB-F cells showed an increased G2 arrest and an inhibition of centrosome overduplication following irradiation mediated by the Rho-kinase, strongly suggesting that RhoB-F may control centrosome overduplication during the G2 arrest after irradiation. Overall, our results for the first time clearly implicate farnesylated RhoB as a crucial protein in mediating cellular resistance to radiation-induced nonapoptotic cell death.

Genotyping of a family with a novel deleterious DPYD mutation supports the pretherapeutic screening of DPD deficiency with dihydrouracil/uracil ratio.

Despite the growing evidence that dihydropyrimidine dehydrogenase deficiency (DPD, encoded by the DPYD gene) confers a higher risk of developing severe toxicity, most patients are not screened for DPD deficiency before fluoropyrimidine treatment. We report here the genetic and phenotypic analyses of DPD in a family related to a patient who died after a first cycle of 5-fluorouracil and in 15 additional retrospective patients having a partial DPD deficiency (as measured by plasma dihydrouracil/uracil ratio). The patient with lethal toxicity was found to be a compound heterozygote for two DPYD mutations: a novel 8-bp duplication (c.168_175dupGAATAATT, p.Phe59Ter) and c.1679T>G (Ile560Ser). The patient's dihydrouracil/uracil ratio indicates complete DPD deficiency. The novel mutation was found in two members of the patient's family. Deleterious DPYD mutations were identified in 9 out of the 15 patients. The relationship between genotype and dihydrouracil/uracil values in the 22 patients of the present study was significant (P = 0.01).

[Isotretinoin: compliance with recommendations in childbearing women]. / Isotrétinoïne: suivi de l'application des recommandations des prescriptions chez les femmes en âge de procréer.

INTRODUCTION: The aim of this survey is to ascertain if the incidence of isotretinoin exposed pregnancies was reduced by the late recommendations of prescription and issue (AMM modification on 06/08/2001 and 25/09/2001). METHODS: All isotretinoin exposed pregnancies registered by the French Regional Drug Monitoring Centres, the Information Centre for Teratogenic Agents and Roche (Roaccutane), Pierre Fabre (Curacné Gé) and Expanscience (Procuta Gé) laboratories, from January 1st, 1999 to December 31st, 2002, were analysed. Enforcement of the strengthening of isotretinoin prescription recommendations was analysed on a sample of 68 prescriptions from 45 pharmacies throughout France. RESULTS: In 4 years, 103 isotretinoin exposed pregnancies (Roaccutane 97 p. 100, Curacné(R) Gé 3 p. 100) during teratogenic risk period, were registered. Pregnancy started less than one month after isotretinoin stopping (37 p. 100), during the treatment (43 p. 100), or was in progress when the treatment was initiated (20 p. 100). The reason of the 22 lacking contraception was known 12 times, i.e. an absence of prescription (6 times), a refusal to take a prescribed contraception (3 times) and a self-medication (3 times). Among the 71 pregnancies whose contraceptive status is known, 48 p. 100 could had been avoided if recommendations had been followed (pregnancies due to a premature stopping or an absence of contraception). The issue of pregnancies is a voluntary termination in 60 cases (87 p. 100). Malformations frequency is 25 p. 100. Incidence of isotretinoin exposed pregnancies remained stable, 0.26/1000 treated women (vs 0.34 after 2001's AMM modifications). Of 68 prescriptions studied, 23 (24 p. 100) carried all the legal warnings, which is close to the previous survey's results. Contraception was in accordance with the recommendations in 78 p. 100 of cases and women learned and applied information given in 38 p. 100 of cases. At last, only 6 patients (9 p. 100) have both a correctly written prescription, a contraception and a time between the pregnancy test date and prescription and issue dates, in accordance with the licence and have had a correct information and understood it. Regarding the previous survey, pregnancy test before treatment was more often prescribed (96 p. 100 vs 88 p. 100). On the other hand, less women knew the necessity to keep on taking contraception one month after isotretinoin stopping (82 p. 100 vs 93 p. 100). CONCLUSION: Despite 3 successive isotretinoin prescription and issue recommendations strengthening in childbearing women, pregnancies can't be totally avoided. Bad compliance concerns the prescription and/or an incomplete or not understood information by the patient who does not scrupulously apply the care and contraception agreement. However, this study does not allow to assess the proportion of issued prescriptions despite their non-accordance with the licence criteria. The National Commission of Pharmacovigilance did not like to limit isotretinoin prescription to dermatologists only. It estimates that the administrative authority must intensify information by dermatologists, general practitioners and pharmacists, about measures to take to avoid an exposure to isotretinoin during pregnancy.