A "chamber of errors" adaptation to assess pharmaceutical assistants' knowledge in chemotherapy preparation.

French preparation guidelines state that pharmacy staff who manipulate cytotoxic drugs have to follow specific training. In order to assess the pharmaceutical assistants' skills and knowledge, we developed a "Cytotoxic Preparation Centralized Unit (CPCU) of errors," derived from the Canadian concept of "Chamber of horrors." A table listing 20 mistakes to track down was created and each pharmaceutical assistant spent 20 min in the "CPCU of errors" with the pharmacist, who wrote down the spotted mistakes in real time. Among the 21 trained pharmaceutical assistants, 15 were evaluated. On average, 11.9 mistakes on 20 were detected. The lowest score was 7 spotted errors on 20 and the highest was 16 on 20. Those results should be qualified depending on pharmaceutical assistants' years of experience in the preparation of chemotherapy. Those results may be explained by the way the role-playing was conducted. The simulation was not conducted during an actual preparation using the usual equipment. One of the major obstacles was the difficulty to clear some time for this project because its realization required a full-time pharmacist and the referring pharmaceutical assistant in addition to the evaluated pharmaceutical assistants. Overall, the staff feedback was positive and the role-playing led to a reminder of theoretical knowledge and the good use of some devices. It would be interesting to develop this type of project through a regional oncology network to create a medium that can be used by other hospitals.

Physicochemical stability of norepinephrine bitartrate in polypropylene syringes at high concentrations for intensive care units.

OBJECTIVES: Norepinephrine is usually used in emergency situations such as in intensive care units (ICUs) for the restoration of blood pressure. The objective was to study the stability of highly-concentrated solutions of norepinephrine at 0.50mg/mL and 1.16mg/mL, diluted in glucose 5% (G5%) in polypropylene syringes, protected or not from light, up to 48h. MATERIALS AND METHODS: Chemical stability was analysed by high-performance liquid chromatography coupled to photodiode array detection at each time of the analysis. The method was validated according to the International Conference on Harmonisation Q2(R1). Physical stability was evaluated by visual and subvisual inspection. Three syringes for each condition were prepared. At each time of the analysis, three samples were analysed for each syringe. pH values were evaluated at each moment of the analysis. RESULTS: Solutions of norepinephrine at 0.50 and 1.16mg/mL, diluted in G5%, with or without protection from light, retained more than 95.0% of the initial concentration after a 48-hour storage at 20-25°C. No visual and subvisual modification occured during the stability study. No degradation product appearing during the stressed degradation was observed during the study but an additional peak with a relative retention at 0.66 was observed and constant. This peak was identified as 5-hydroxymethylfurfural, a degradation product of glucose. CONCLUSION: Norepinephrine diluted in G5% at 0.50mg/mL and 1.16mg/mL was physically and chemically stable over a period of 48hours at room temperature. These stability data of highly concentrated solutions provide additional knowledge to assist intensive care services in daily practice.

First international database of research teams in stability and compatibility of medications: An additional function in the Stabilis® database.

A new function on the international database Stabilis® has been created: a database of research teams involved in stability and compatibility studies of drugs. The first part is descriptive. For each team, the list of publications and of molecules studied can be edited. A search function can extract data by using different criteria like the drug, galenic form, pharmacological class, molecule form (small molecule, monoclonal antibody), country, author and years of publications. The second part of the new function is interactive and allows Stabilis® users to suggest stability studies. After examination by collaborators of Stabilis®, the suggestions are rejected, modified or accepted, published on the website and available for all research teams. The United States of America (USA) was the main country in the 1990s but today, Europe has 67 active teams which publish stability studies and 50 for USA. Europe is now the more active zone in term of number of publications. Antibiotics and cytotoxics are the main pharmacological classes studied. Monoclonal antibodies are studied by few teams due to the complexity of the methodology to study the stability of these drugs. This database can facilitate the communication between research teams and users.

[Study impacto: Descriptive analyzis of pharmacist's clinical practice in onco-hematology]. / Étude impacto : analyse descriptive des pratiques de pharmacie clinique en cancérologie.

Pharmaceutical analyses of chemotherapy prescriptions by hospital pharmacists are activities codified by regulation and rules (bon usage). The involvement of the pharmacists in clinical pharmacy activities in the oncology setting is not clearly identified, justifying the development of a mapping of these activities from a questionnaire addressed to the professionals. One hundred and seven centers have participated to this study at the national level (overall participation rate of 32.4%). More than 95% of them used a computerized ordering system and three quarter of them submit the introduction of new compounds to an analysis by the drug therapeutic committee. Prescription analysis allowed detecting around 2% of errors from the current prescription. Clinical pharmacist participates to tumor boards of onco-hematology (RCP) at a level of 46% for senior pharmacist and 42% for junior pharmacist. This involvement in the RCP allowed anticipating protocol's modification and temporary used authorization. Ninety-two percent of the senior pharmacists estimate that they highlight the risk of no reimbursement for prescription out of the guideline during RCP, resulting to a modification of the prescription for 40% of them. This level of intervention is lower with respectively 64% and 10% for the juniors. This study underlines the expert value of the clinical pharmacist dedicated to oncology setting in pre and post analysis prescriptions. It could be targeted by a prospective analysis of both clinical and pharmacoeconomics impact of these interventions.

A human homologue of the Drosophila eyes absent gene underlies branchio-oto-renal (BOR) syndrome and identifies a novel gene family.

A candidate gene for Branchio-Oto-Renal (BOR) syndrome was identified at chromosome 8q13.3 by positional cloning and shown to underlie the disease. This gene is a human homologue of the Drosophila eyes absent gene (eya), and was therefore called EYA1. A highly conserved 271-amino acid C-terminal region was also found in the products of two other human genes (EYA2 and EYA3), demonstrating the existence of a novel gene family. The expression pattern of the murine EYA1 orthologue, Eya1, suggests a role in the development of all components of the inner ear, from the emergence of the otic placode. In the developing kidney, the expression pattern is indicative of a role for Eya1 in the metanephric cells surrounding the 'just-divided' ureteric branches.

Mutations in the skeletal muscle alpha-actin gene in patients with actin myopathy and nemaline myopathy.

Muscle contraction results from the force generated between the thin filament protein actin and the thick filament protein myosin, which causes the thick and thin muscle filaments to slide past each other. There are skeletal muscle, cardiac muscle, smooth muscle and non-muscle isoforms of both actin and myosin. Inherited diseases in humans have been associated with defects in cardiac actin (dilated cardiomyopathy and hypertrophic cardiomyopathy), cardiac myosin (hypertrophic cardiomyopathy) and non-muscle myosin (deafness). Here we report that mutations in the human skeletal muscle alpha-actin gene (ACTA1) are associated with two different muscle diseases, 'congenital myopathy with excess of thin myofilaments' (actin myopathy) and nemaline myopathy. Both diseases are characterized by structural abnormalities of the muscle fibres and variable degrees of muscle weakness. We have identified 15 different missense mutations resulting in 14 different amino acid changes. The missense mutations in ACTA1 are distributed throughout all six coding exons, and some involve known functional domains of actin. Approximately half of the patients died within their first year, but two female patients have survived into their thirties and have children. We identified dominant mutations in all but 1 of 14 families, with the missense mutations being single and heterozygous. The only family showing dominant inheritance comprised a 33-year-old affected mother and her two affected and two unaffected children. In another family, the clinically unaffected father is a somatic mosaic for the mutation seen in both of his affected children. We identified recessive mutations in one family in which the two affected siblings had heterozygous mutations in two different exons, one paternally and the other maternally inherited. We also identified de novo mutations in seven sporadic probands for which it was possible to analyse parental DNA.

SFPO and ESOP recommendations for the practical stability of anticancer drugs: an update.

The recommendations for the practical stability of anticancer drugs published in 2010 by the French Society of Hospital Pharmacists (SFPO) and the European Society of Oncology Pharmacists (ESOP) have been updated. Ten new molecules have been included (asparaginase, azacitidine, bevacizumab, clofarabine, eribuline mesylate, folinate sodium, levofolinate calcium, nelarabine, rituximab, temsirolimus).

Stability study of amiodarone hydrochloride in capsules for paediatric patients using a high-performance liquid chromatography method.

Amiodarone hydrochloride, a class III antiarrhythmic agent, shows ß blocker-like and potassium channel blocker-like actions on the sinuatrial and atrioventricular nodes. It is given by mouth in the treatment of all forms of atrial, junctional and ventricular arrhythmias. Capsules for paediatric patients are not commercially available and must be prepared in the pharmacy department. The aim of the study was to evaluate the stability of amiodarone hydrochloride at different dosages, 10, 60 and 100 mg, in capsules for paediatric patients stored in three packages under dark conditions and at room temperatures over one year. At different intervals during the storage period, amiodarone hydrochloride concentration was tested using a high-performance liquid chromatography (HPLC) method. Amiodarone hydrochloride content remained greater than 95% of the initial concentration in all capsules at all dosages. The 10, 60 and 100 mg amiodarone paediatric capsules were stable for one year when stored in the three packages at ambient temperature and under dark conditions.

Guidelines for the practical stability studies of anticancer drugs: a European consensus conference.

Stability studies performed by the pharmaceutical industry are only designed to fulfill licensing requirements. Thus, post-dilution or -reconstitution stability data are frequently limited to 24h only for bacteriological reasons regardless of the true chemical stability which could, in many cases, be longer. In practice, the pharmacy-based centralized preparation may require infusions to be made several days in advance to provide, for example, the filling of ambulatory devices for continuous infusions or batch preparations for dose banding. Furthermore, a non-justified limited stability for expensive products is obviously very costly. Thus, there is a compelling need for additional stability data covering practical uses of anticancer drugs. A European conference consensus was held in France, May 2010, under the auspices of the French Society of Oncology Pharmacy (SFPO) to propose adapted rules on stability in practical situations and guidelines to perform corresponding stability studies. For each anticancer drug, considering their therapeutic index, the pharmacokinetics/pharmacodynamics (PK/PD) variability, specific clinical use and risks related to degradation products, the classical limit of 10% of degradation can be inappropriate. Therefore, acceptance limits must be clinically relevant and should be defined for each drug individually. Design of stability studies has to reflect the different needs of the clinical practice (preparation for the week-ends, outpatient transportations, implantable devices, dose banding…). It is essential to use validated stability-indicating methods, separating degradation products being formed in the practical use of the drug. Sequential temperature designs should be encouraged to replicate problems seen in daily practice such as rupture of the cold-chain or temperature-cycling between refrigerated storage and ambient in-use conditions. Stressed conditions are recommended to evaluate not only the role of classical variables (pH, temperature, light) but also the mechanical stress. Physical stability such as particles' formation should be systematically evaluated. The consensus conference focused on the need to perform more studies on the stability of biotherapies, including a minimum of three complementary separating methods and a careful evaluation of submicron aggregates. The determination of the biological activity of proteins could be also useful. A guideline on the practical stability of anticancer drugs is proposed to cover current clinical and pharmaceutical practice. It should contribute to improved security of use, optimization of centralized handling and reduced costs. Finally, we have attempted to establish a new drug stability paradigm based on practical clinical needs, to complement regulatory guidelines which are essentially orientated to the stability of manufactured drugs.

Mutation update for the CSB/ERCC6 and CSA/ERCC8 genes involved in Cockayne syndrome.

Cockayne syndrome is an autosomal recessive multisystem disorder characterized principally by neurological and sensory impairment, cachectic dwarfism, and photosensitivity. This rare disease is linked to mutations in the CSB/ERCC6 and CSA/ERCC8 genes encoding proteins involved in the transcription-coupled DNA repair pathway. The clinical spectrum of Cockayne syndrome encompasses a wide range of severity from severe prenatal forms to mild and late-onset presentations. We have reviewed the 45 published mutations in CSA and CSB to date and we report 43 new mutations in these genes together with the corresponding clinical data. Among the 84 reported kindreds, 52 (62%) have mutations in the CSB gene. Many types of mutations are scattered along the whole coding sequence of both genes, but clusters of missense mutations can be recognized and highlight the role of particular motifs in the proteins. Genotype-phenotype correlation hypotheses are considered with regard to these new molecular and clinical data. Additional cases of molecular prenatal diagnosis are reported and the strategy for prenatal testing is discussed. Two web-based locus-specific databases have been created to list all identified variants and to allow the inclusion of future reports (www.umd.be/CSA/ and www.umd.be/CSB/).

Delineation of 15q13.3 microdeletions.

The increasing use of array-comparative genomic hybridization (array-CGH) to identify copy number variations (CNVs) in patients with developmental delay (DD), mental retardation and/or dysmorphic features has allowed the recent recognition of numerous genomic imbalances, including the 15q13.3 microdeletion. Patients with this microdeletion generally present with relatively consistent breakpoints at BP4 and BP5, which include the CHRNA7 gene. About 100 index cases have been reported since the first publication in 2008. This large number of patients ascertained through highly variable samples has been necessary to describe the full phenotypic spectrum of this microdeletion, ranging from mental retardation with dysmorphic features, epilepsy, neuropsychiatric disturbances with or without cognitive impairment to complete absence of anomalies. Here, we describe a collaborative study reporting a new cohort of 12 index patients and 13 relatives carrying a heterozygous BP4-BP5 microdeletion out of a series of 4625 patients screened by array-CGH for DD. We confirm the clinical expressivity of the disease as well as the incomplete penetrance in seven families. We showed through a review of the literature that males are more likely to be symptomatic. Sequence analysis of CHRNA7 yielded no data to support the unmasking of recessive variants as a cause of phenotypic variability. We also report the first patient carrying a 15q13.3 homozygous microdeletion inherited from both parents. He had severe epileptic encephalopathy with retinopathy, autistic features and choreoathetosis. Besides the classical approximately 1.5 Mb BP4-BP5 microdeletion, we also describe three index patients and two relatives with a smaller 500 kb microdeletion, including the CHRNA7 gene.

Confirmation of TFAP2A gene involvement in branchio-oculo-facial syndrome (BOFS) and report of temporal bone anomalies.

Branchio-oculo-facial syndrome (BOFS) is an autosomal-dominant condition characterized by three main features, respectively: branchial defects, ocular anomalies, and craniofacial defects including cleft lip and/or palate (CL/P). We report on one family with three affected, and two sporadic cases that have been found to carry missense mutations in the newly reported BOFS gene: TFAP2A. This report confirms the involvement of this transcription factor in this developmental syndrome with clinical variability. Moreover, we present CT scan temporal bone anomalies in the familial cases, related to branchial arch defects, highlighting the importance of radiological investigations for differential diagnosis.

Optical near-field excitations on plasmonic nanoparticle-based structures.

We investigate optical excitations on single silver nanospheres and nanosphere composites with the Finite Difference Time Domain (FDTD) method. Our objective is to achieve polarization control of the enhanced local field, pertinent to SERS applications. We employ dimer and quadrumer structures, which can display broadband and highly confined near-field-intensity enhancement comparable to or exceeding the resonant value of smaller sized isolated spheres. Our results demonstrate that the polarization of the enhanced field can be controlled by the orientation of the multimers in respect to the illumination, rather than the illumination itself. In particular, we report cases where the enhanced field shares the same polarization with the exciting field, and cases where it is predominantly perpendicular to the source field. We call the later phenomenon depolarized enhancement. Furthermore, we study a realizable nanolens based on a tapered self-similar silver nanosphere array. The time evolution of the fields in such structures show conversion of a diffraction limited Gaussian beam to a focused spot, through sequential coupling of the nano-array spheres' Mie-plasmons. For a longitudinally excited nanolens design we observed the formation of an isolated focus with size about one tenth the vacuum wavelength. We believe such nanolens will aid scanning near-field optical microscopy (SNOM) detection and the excitation of surface plasmon based guiding devices.

Grafting and polymer formation on silicon from unsaturated Grignards: II. Aliphatic precursors.

Anodic decomposition of a vinylmagnesium halide or an ethynylmagnesium halide at a surface-hydrogenated silicon electrode leads to the formation of polymeric layers covalently anchored to the silicon surface. These layers have been characterized using spectroellipsometry and photoluminescence, infrared, and X-ray photoelectron spectroscopy. In the case of vinyl precursors, it appears that the multiple bonds are largely broken in the process. In the case of ethynyl, the layer formation rate is much higher for the chloride than for the bromide. The obtained polymer appears as a saturated skeleton bearing halide and unsaturated ethynyl groups. Furthermore, it appears that the solvent may be attacked by the ethynyl radicals leading to contamination of the polymer by solvent fragments, an effect that can largely be avoided by using appropriate solvents. The reaction pathways are discussed.

Characterization of the surface physico-chemistry of plasticized PVC used in blood bag and infusion tubing.

Commercial infusion tubing and blood storage devices (tubing, blood and platelets bags) made of plasticized PVC were analyzed by spectroscopic, chromatographic and microscopic techniques in order to identify and quantify the additives added to the polymer (lubricants, thermal stabilizers, plasticizers) and to put into evidence their blooming onto the surface of the devices. For all the samples, deposits were observed on the surface but with different kinds of morphologies. Ethylene bis amide lubricant and metallic stearate stabilizers were implicated in the formation of these layers. In contact with aqueous media, these insoluble deposits were damaged, suggesting a possible particulate contamination of the infused solutions.

Color-selecting reflectors inspired from biological periodic multilayer structures.

We propose a semi-infinite 1-D photonic crystal approach for designing artificial reflectors which aim to reproduce color changes with the angle of incidence found in biological periodic multilayer templates. We show that both the dominant reflected wavelength and the photonic bandgap can be predicted and that these predictions agree with exact calculations of reflectance spectra for a finite multilayer structure. In order to help the designer, the concept of spectral richness of angle-tuned color-selecting reflectors is introduced and color changes with angle are displayed in a chromaticity diagram. The usefulness of the photonic crystal approach is demonstrated by modelling a biological template (found in the cuticle of Chrysochora vittata beetle) and by designing a bio-inspired artificial reflector which reproduces the visual aspect of the template. The bioinspired novel aspect of the design relies on the strong unbalance between the thicknesses of the two layers forming the unit cell.

Grafting and polymer formation on silicon from unsaturated Grignards: I-aromatic precursors.

Anodic decomposition of a phenylmagnesium halide at a surface-hydrogenated silicon electrode leads to formation of polymeric layers covalently anchored to the silicon surface. These layers have been characterized using spectroellipsometry, photoluminescence, infrared, and X-ray photoelectron spectroscopies. The phenyl ring appears preserved in the process, and the polymer formed is a polyphenylene. Contamination by aliphatic groups from the solvent may be minimized by using a solvent resistant to hydrogen abstraction by the phenyl radicals. Regioselectivity of the branching may be oriented to the para form by using 4-chlorophenylmagnesium bromide as the precursor.

The design of cationic lipids for gene delivery.

Synthetic gene delivery vectors are gaining increasing importance in gene therapy as an alternative to recombinant viruses. Among the various types of non-viral vectors, cationic lipids are especially attractive as they can be prepared with relative ease and extensively characterised. Further, each of their constituent parts can be modified, thereby facilitating the elucidation of structure-activity relationships. In this forward-looking review, cationic lipid-mediated gene delivery will mainly be discussed in terms of the structure of the three basic constituent parts of any cationic lipid: the polar headgroup, hydrophobic moiety and linker. Particular emphasis will be placed on recent advances in the field as well as on our own original contributions. In addition to reviewing critical physicochemical features (such as headgroup hydration) of monovalent lipids, the use of headgroups with known nucleic-acid binding modes, such as linear and branched polyamines, aminoglycosides and guanidinium functions, will be comprehensively assessed. A particularly exciting innovation in linker design is the incorporation of environment-sensitive groups, the intracellular hydrolysis of which may lead to more controlled DNA delivery. Examples of pH-, redox- and enzyme-sensitive functional groups integrated into the linker are highlighted and the benefits of such degradable vectors can be evaluated in terms of transfection efficiency and cationic lipid-associated cytotoxicity. Finally, possible correlations between the length and type of hydrophobic moiety and transfection efficiency will be discussed. In conclusion it may be foreseen that in order to be successful, the future of cationic lipid-based gene delivery will probably require the development of sophisticated virus-like systems, which can be viewed as "programmed supramolecular systems" incorporating the various functions required to perform in a chronological order the different steps involved in gene transfection.

A "green" strategy to construct non-covalent, stable and bioactive coatings on porous MOF nanoparticles.

Nanoparticles made of metal-organic frameworks (nanoMOFs) attract a growing interest in gas storage, separation, catalysis, sensing and more recently, biomedicine. Achieving stable, versatile coatings on highly porous nanoMOFs without altering their ability to adsorb molecules of interest represents today a major challenge. Here we bring the proof of concept that the outer surface of porous nanoMOFs can be specifically functionalized in a rapid, biofriendly and non-covalent manner, leading to stable and versatile coatings. Cyclodextrin molecules bearing strong iron complexing groups (phosphates) were firmly anchored to the nanoMOFs' surface, within only a few minutes, simply by incubation with aqueous nanoMOF suspensions. The coating procedure did not affect the nanoMOF porosity, crystallinity, adsorption and release abilities. The stable cyclodextrin-based coating was further functionalized with: i) targeting moieties to increase the nanoMOF interaction with specific receptors and ii) poly(ethylene glycol) chains to escape the immune system. These results pave the way towards the design of surface-engineered nanoMOFs of interest for applications in the field of targeted drug delivery, catalysis, separation and sensing.