[Detection of B19 parvovirus in plasma pools before solvent-detergent treatment of plasma: AFSSAPS and EFS Aquitaine-Limousin's experience]. / Détection du génome viral du parvovirus B19 dans les pools de plasmas servant à la préparation du plasma frais congelé traité pour viro-atténuation par solvant-détergent : expérience de l'Afssaps et de l'EFS Aquitaine-Limousin.

Since 1998, the Aquitaine-Limousin branch of the French Blood Institute has set up a parvovirus B19 (PV B19) systematic screening on each unit of plasma to be treated by solvent-detergent procedure for virus inactivation. Parvovirus B19 nucleic acid systematic testing in plasma pools became mandatory since 2005 (European monograph "Human plasma" - pooled and treated for virus inactivation). The French competent state authority (AFSSAPS) has decided to introduce this test as a part of the external quality control of labile blood products. This process is related to the harmonization of quality control practice realised on blood products in Europe even if the human plasma pooled and treated for virus inactivation by solvent-detergent is considered in France as a blood labile component. Implementation of this test required a validation step and a close cooperation between AFSSAPS and Aquitaine-Limousin blood transfusion centre. Validation consisted in perfecting a semi-quantitative, real-time nucleic acid testing method with automated extraction. This collaborative study leads us to control 1642 plasma pools. All the results were under the threshold of 10,0 IU/microL. AFSSAPS's results were in agreement with those of Aquitaine-Limousin's blood transfusion center who carry out the parvovirus B19 screening both on fresh frozen plasma units composing the pool and on plasma pools.

Challenging near infrared spectroscopy discriminating ability for counterfeit pharmaceuticals detection.

This study was initiated by the laboratories and control department of the French Health Products Safety Agency (AFSSAPS) as part of the fight against the public health problem of rising counterfeit and imitation medicines. To test the discriminating ability of Near InfraRed Spectroscopy (NIRS), worse cases scenarios were first considered for the discrimination of various pharmaceutical final products containing the same Active Pharmaceutical Ingredient (API) with different excipients, such as generics of proprietary medicinal products (PMP). Two generic databases were explored: low active strength hard capsules of Fluoxetine and high strength tablets of Ciprofloxacin. Then 4 other cases involving suspicious samples, counterfeits and imitations products were treated. In all these cases, spectral differences between samples were studied, giving access to API or excipient contents information, and eventually allowing manufacturing site identification. A chemometric background is developed to explain the optimisation methodology, consisting in the choices of appropriate pretreatments, algorithms for data exploratory analyses (unsupervised Principal Component Analysis), and data classification (supervised cluster analysis, and Soft Independent Modelling of Class Analogy). Results demonstrate the high performance of NIRS, highlighting slight differences in formulations, such as 2.5% (w/w) in API strength, 1.0% (w/w) in excipient and even coating variations (<1%, w/w) with identical contents, approaching the theoretical limits of NIRS sensitivity. All the different generic formulations were correctly discriminated and foreign PMP, constituted of formulations slightly different from the calibration ones, were also all discriminated. This publication addresses the ability of NIRS to detect counterfeits and imitations and presents the NIRS as an ideal tool to master the global threat of counterfeit drugs.

Trace determination of ethyl methanesulfonate in Viracept 250 mg tablets by gas chromatography-mass spectrometry.

Ethyl methanesulfonate (EMS) is a potential human mutagenic and carcinogenic compound which has been found by Roche laboratories in nelfinavir mesylate, the active pharmaceutical ingredient of Viracept. In order to verify the quality of the medicinal product, a gas chromatographic method using mass spectrometry detection was developed for the trace analysis of EMS in Viracept 250 mg tablets from Roche laboratories. Combined with suitable sample preparation including a liquid/liquid extraction this method allows the EMS quantification with a reporting limit of 5 ppm. The extract is injected on a gas chromatographic system with a CP624-CB capillary column. Selected Ion Monitoring mode was used for the EMS quantification. Some validation elements of the method are reported. The validation study was performed over a range from 5 ppm to 100 ppm.

In vitro pyrogenicity of Gram-positive bacteria--validation of the kit using fresh human whole blood.

The two conventional tests to detect pyrogen contaminants in injectable pharmaceutical drugs are the Rabbit Model and the Limulus amoebocyte lysate (LAL) test. To replace these models, a new system on human whole blood is developed, using the release of Interleukin 1 beta (IL1beta) after cell stimulation with gram-positive and gram negative pyrogens. The purpose of this study was to validate the ENDOSAFE-IPT kit using the quantitative ELISA enzyme immunoassay. The assay is divided into two parts: blood cell stimulation with Lipopolysaccharides (LPS) and Lipoteichoic acid (LTA) and quantitation of IL1beta using the ELISA method. In each assay, blood from a particular donor were stimulated with the Endotoxin Standard, and with a sample of a commercial antibiotic preparation (Clavulanic acid/Ticarcillin) spiked with the Endotoxin Standard. LTA from Bacillus subtilis and a sample of diphtheria toxoid were also used. At least, six assays were tested. A polynomial regression of the Endotoxin Standard series showed a correlation coefficient greater than 0.99. The spiked antibiotic sample recoveries were 50-121%. The LTA quantitation limit was 0.1 microg/ml and the range of detection of pyrogens from Gram positive diphtheria toxoid was 0.77 to 2.5 EEU/ml. The IL1beta production varied markedly between donors. However the coefficient of variation was less than 20 % intra-assay. In conclusion, the ENDOSAFE-IPT kit can be used for the quantitative and qualitative detection of pyrogens from Gram negative and Gram positive bacteria.

[Cosmetovigilance: a French pharmacovigilance system for cosmetics developed by the French health products safety agency. A proposal for the future]. / Le système de cosmétovigilance français géré par l'Agence française de sécurité sanitaire des produits de santé. Une proposition pour le futur.

Since 1999, the French health products safety agency (Afssaps) has been responsible for the safety of cosmetic products. Attempts are underway to set up a French post-marketing surveillance system for cosmetics and a working group specially in charge of setting this up has been created. The "cosmetovigilance" system has the three following objectives: public health concerns about "at risk" ingredients in cosmetic products: animal derivatives, glycol ethers, aluminium, camphor, etc; notification of adverse reactions: creation of standard reporting forms of adverse reactions; follow-up of laboratory quality controls: emergency and routine controls. The "cosmetovigilance" system is not only restricted to the study of adverse reactions occurring with cosmetic products. It will also address public health issues about ingredients and will rely on laboratory controls. The aim of the "cosmetovigilance" system is to investigate but also to prevent the risk of adverse reactions.

Central action of 5-HT3 receptor ligands in the regulation of sleep-wakefulness and raphe neuronal activity in the rat.

Anxiolytic drugs, such as the benzodiazepines and the azapirones (ipsapirone, gepirone, buspirone), are well known to affect states of vigilance and to decrease the firing rate of serotoninergic neurones within the dorsal raphe nucleus in rats. In order to examine whether the newly developed 5-HT3 antagonists with potential anxiolytic properties act through similar mechanisms, the effects of several of such antagonists: MDL 72222, ICS 205-930, ondansetron and/or zacopride on both sleep-wakefulness and the discharge of serotoninergic neurones within the dorsal raphe nucleus were investigated in rats. When tested in a wide range of doses (0.05-10 mg/kg, i.p.), none of these drugs significantly affected the states of vigilance, except ondansetron, at 0.1 mg/kg, which increased paradoxical sleep for the first 2 hr after administration and MDL 72222, at 10 mg/kg, which reduced both paradoxical and slow wave sleep and increased wakefulness for the same initial period after treatment. In vivo, in chloral hydrate anaesthetized rats, as well as in vitro, in slices of brain stem, none of the 5-HT3 antagonists tested affected the firing rate of serotoninergic neurones. Similarly, no change in the electrical activity of serotoninergic neurones could be evoked in vitro by superfusion of the tissue with the 5-HT3 agonists, phenylbiguanide (10 microM) and 2-methyl-5-HT (1 microM). At a larger concentration (10 microM), the latter compound reduced the neuronal discharge probably through the stimulation of somatodendritic 5-HT1A autoreceptors since this effect, as that of ipsapirone, could be prevented by 10 microM l-propranolol. Comparison of these data with those obtained with benzodiazepines and 5-HT1A agonists of the azapirone series, supports the concept that different mechanisms are responsible for the anxiolytic-like properties of 5-HT3 agonists, compared to those of other anxiolytic drugs.

Antidepressant-like effects of buspirone mediated by the 5-HT1A post-synaptic receptors in the learned helplessness paradigm.

The 5-HT1A receptor agonists buspirone and 8-OH-DPAT have strong effects on serotoninergic systems. Mediated by both pre- and post-synaptic 5-HT1A receptors, these pharmacological effects might predict both antidepressant and antianxiety activities. In animal models sensitive to antidepressant drugs, the 5-HT1A agonists administered i.p. have been shown to mimic the behavioral effects of tricyclics. In the present study, the learned helplessness paradigm was used to assess the possible role of pre- or post-synaptic 5-HTIA receptors in this effect. The ability of buspirone compared with 8-OH-DPAT to reduce helpless behavior was investigated after local microinjections (0.1 or 1.0 micrograms in 0.5 microliters) into the raphe nuclei or into the septum. The results indicate that microinjections of buspirone or 8-OH-DPAT into the raphe nuclei did not reverse helpless behavior; in contrast, microinjections of both 5-HTIA agonists into the septum reverse helpless behavior. These results suggest that antidepressant-like properties of buspirone and 8-OH-DPAT may be mediated, in this test, by the post-synaptic 5-HTIA receptors through functional enhancement of the 5-HT transmission.

[Experimental study of changes in the pressure of the posterior segment of the eye under general anesthesia. Consequences for surgery of the anterior segment]. / Etude expérimentale des variations de la pression du segment postérieur de l'oeil sous anesthésie générale. Conséquence pour la chirurgie du segment antérieur.

Animal studies were conducted to compare variations in intraocular (IOP) and posterior segment pressure (PSP) during general anesthesia to assess the role of PSP in the development of anesthesia for ophthalmological procedures. Anesthetic agents appear to have a marked effect on IOP during operations involving opening of hypertonic globe or examinations of children under general anesthesia, but their action on IOP has no significance during procedures requiring opening of the anterior chamber because of alteration of aqueous humor physiology. The PSP, defined as pressure in the posterior segment when the anterior segment is at atmospheric pressure, is the main factor affecting surgical conditions, a rise in PSP possibly resulting in typical complications of cataract surgery but having beneficial effects in corneal grafts for example. Posterior segment pressure cannot be studied in humans and an experimental model using rabbits under artificial ventilation following tracheotomy after general anesthesia was developed. Pressure gauges recorded arterial and central venous pressures and were connected to needles inserted in both eyes to monitor IOP and PSP, the latter from a needle passed into the anterior chamber through the cornea, which was incised over the needle to enable permanent drainage of aqueous humor. All pressures were recorded simultaneously and no correlations were observed between IOP and PSP after pentobarbitone, neosynephrine, succinylcholine, or asphyxia (interruption of ventilation and curarization). These findings suggest that IOP is not a valid measurement for assessment of anesthetic techniques, whereas PSP provides a better guideline for development of ophthalmological anesthesia.