Diclofenac Loaded Lipid Nanovesicles Prepared by Double Solvent Displacement for Skin Drug Delivery.

PURPOSE: Herein, we detail a promising strategy of nanovesicle preparation based on control of phospholipid self-assembly: the Double Solvent Displacement. A systematic study was conducted and diclofenac as drug model encapsulated. In vitro skin studies were carried out to identify better formulation for dermal/transdermal delivery. METHODS: This method consists in two solvent displacements. The first one, made in a free water environment, has allowed triggering a phospholipid pre-organization. The second one, based on the diffusion into an aqueous phase has led to liposome formation. RESULTS: Homogeneous liposomes were obtained with a size close to 100 nm and a negative zeta potential around -40 mV. After incorporation of acid diclofenac, we obtained nanoliposomes with a size between 101 ± 45 and 133 ± 66 nm, a zeta potential between 34 ± 2 and 49 ± 3 mV, and the encapsulation efficiency (EE%) was between 58 ± 3 and 87 ± 5%. In vitro permeation studies showed that formulation with higher EE% dispayed the higher transdermal passage (18,4% of the applied dose) especially targeting dermis and beyond. CONCLUSIONS: Our results suggest that our diclofenac loaded lipid vesicles have significant potential as transdermal skin drug delivery system. Here, we produced cost effective lipid nanovesicles in a merely manner according to a process easily transposable to industrial scale. Graphical Abstract ᅟ.

[Analysis of the question-answer activity of a hospital pharmacy. Example of the handling of drug interactions]. / Analyse de l'activité question-réponse d'une pharmacie à usage intérieur. Exemple de la gestion des interactions médicamenteuses.

INTRODUCTION: The medical care of patients generates questions among healthcare professionals. Some will necessitate an advanced research. The hospital pharmacist is at the interface between prescribers, caregivers and the medicines and is requested to answer these requests. Studies conducted in other countries showed that this question-answer activity represents a significant amount of time in daily work. In France, this topic was poorly explored. The objective of our work was to study the volume and the type of questions, the clinical situations, the time required, the medicines implicated and the sources of information used. MATERIALS AND METHODS: A prospective study was conducted in the pharmacy of a university hospital. All the requests answered by the pharmaceutical team, which needed a specific research, analysis and writing of an answer were collected. RESULTS: A hundred and one questions were analyzed, originating from doctors or medicals interns. Almost half concerned drug interactions, and among them, almost a fourth were not mentioned in the Summary of Product Characteristics of the medicines involved. A pharmaceutical advice was provided in 91.5% of the cases. Time dedicated to the research varied between less than 30 minutes and more than 8 hours. DISCUSSION AND CONCLUSION: This study illustrates the question-answer activity of a hospital pharmacy, which is currently not taken into account as an indicator of pharmaceutical activity. A large part concerns analysis and management of drug interactions and requires a significant amount of pharmaceutical time.

Identification and prioritization of bioaccumulable pharmaceutical substances discharged in hospital effluents.

The consumption of pharmaceuticals and their excretion in wastewater is a continuous source of pollution for aquatic ecosystems. In certain cases these compounds are found in the environment at concentrations high enough to cause disturbance in aquatic organisms. Aware of this problem hospitals are giving increasing attention to the nature of their effluents and their impact on the environment, by implementing more efficient effluent management policies. This concern is justified in view of the large volumes of toxic products consumed (detergents, disinfectants, pharmaceuticals, chemical reagents, radioactive elements, etc.). Moreover, these effluents usually do not undergo any specific treatment before being discharged into urban sewage networks. In this article, we present a method for selecting the pharmaceuticals discharged in hospital effluents that have the worst impact on the aquatic ecosystem, primarily based on their bioaccumulation potential. This study focused on the pharmaceuticals consumed at the Hospices Civils de Lyon (HCL), the second largest hospital structure in France (5200 hospital beds). Of the 960 substances consumed in HCL hospitals, a shortlist of 70 substances considered as being potentially bioaccumulable was established. The use of aggravating factors of risk has then led to the final selection of 14 priority compounds. They include 4 compounds consumed in large quantities in HCL hospitals, 6 endocrine disruptors and 4 potentially ecotoxic compounds. For all these compounds, it is now advisable to verify their bioaccumulation potential experimentally and confirm their presence in the environment. In addition, in order to monitor the risk relating to possible contamination of the food chain, it will be necessary to measure accumulated dose levels in species of different trophic levels. Lastly, chronic ecotoxicity tests will permit evaluating the danger and risk that some of these substances may represent for aquatic ecosystems.

[The evolution of pharmaceutical practice within the context of changing health territories: How can social sciences contribute to pharmaceutical education?]. / Evolution de la pratique pharmaceutique en contexte de redéfinition des territoires de la santé : ce que les sciences humaines et sociales peuvent apporter à la formation du pharmacien.

Pharmaceutical practice in France is evolving as presented in the legislation reforming hospitals and regarding patients, health and the territories. Hence, the pharmaceutical "territory" has been reconfigured and requires evolving ideas about pharmaceutical training. At the heart of this paper is the following question: What can social sciences bring to pharmaceutical education? Three levels were considered: patients and their social environment, interaction and coordination management among health professionals and political, economical and social drug regulation systems.

In vitro micronucleus assay with Chinese hamster V79 cells - results of a collaborative study with in situ exposure to 26 chemical substances.

A collaborative study with 10 participating laboratories was conducted to evaluate a test protocol for the performance of the in vitro micronucleus (MN) test using the V79 cell line with one treatment and one sampling time only. A total of 26 coded substances were tested in this study for MN-inducing properties. Three substances were tested by all 10 laboratories and 23 substances were tested by three or four laboratories in parallel. Six aneugenic, 7 clastogenic and 6 non-genotoxic chemicals were uniformly recognised as such by all laboratories. Three chemicals were tested uniformly negative by three laboratories although also clastogenic properties have been reported for these substances. Another set of three clastogenic substances showed inconsistent results and one non-clastogenic substance was found to be positive by one out of three laboratories. Within the study, the applicability of the determination of a proliferation index (PI) as an internal cytotoxicity parameter in comparison with the determination of the mitotic index (MI) was also evaluated. Both parameters were found to be useful for the interpretation of the MN test result with regard to the control of cell cycle kinetics and the mode of action for MN induction. The MN test in vitro was found to be easy to perform and its results were mainly in accordance with results from chromosomal aberration tests in vitro.

4-Chloro-o-phenylenediamine: a 26-week oral (in feed) mutagenicity study in Big Blue mice.

4-Chloro-o-phenylenediamine (4-C-o-PDA) is a liver carcinogen in mice and was found to be weakly mutagenic in the liver of female Big Blue mice after short term treatment. In the present study the test compound was given subchronically in the diet for 26 weeks at doses of 0, 5000 and 10,000 ppm. The corresponding average test substance intake was 2166 mg kg-1 day-1 (males: 1794 mg kg-1 day-1; females: 2539 mg kg-1 day-1) and 4610 mg kg-1 day-1 (males: 3926 mg kg-1 day-1; females 5925 mg kg-1 day-1) at the low and high dose, respectively. After sacrifice, tissues were flash frozen in liquid nitrogen. The lacI mutant frequency in the liver was determined from three male and three female mice per dose group. The genomically integrated transgene was recovered by packaging into lambda phage using Transpack packaging extract (Stratagene, La Jolla, USA) followed by infection of Escherichia coli strain SCS-8. Blue mutant plaques were scored against a background of clear non-mutant plaques. Food consumption decreased initially at 10,000 ppm, while no treatment related effect on food intake was observed at 5000 ppm. Body weight gain was found to be decreased in all treated animals. Absolute and relative liver weight increased in a dose-related manner, but only the latter effect was statistically significant. A clear dose dependent increase in lacI mutant frequencies was observed in the liver of both sexes. The following mutant frequencies (x10(-5)) were observed: 2.73+/-1.01 (males, untreated), 7.24+/-1.50 (females, untreated), 18.91+/-5.30 (5000 ppm, males), 24.91+/-7.58 (5000 ppm, females), 20.47+/-6.68 (10,000 ppm, males) and 36.17+/-14.98 (10,000 ppm, females). It is therefore concluded that 4-C-o-PDA is a strong mutagen in the liver of mice treated subchronically for 26 weeks.

Automatic analysis of the in vitro micronucleus test on V79 cells.

The in vitro micronucleus test is a well established test for early screening of new chemical entities in industrial toxicology. For assessing the clastogenic or aneugenic potential of a test compound, micronucleus induction in cells has been shown repeatedly to be a sensitive and specific parameter. As a measure for numerical and structural chromosome aberrations, the in vitro micronucleus test consists of determining the frequency of micronucleated cells in a representative fraction of cells in a culture. So far, manual counting has been the only method for evaluating microscopic V79 Chinese hamster cell preparations. To replace this tedious and time consuming procedure, a fully automatic system for micronucleus scoring in V79 cells by image analysis has been developed and introduced into the routine genotoxicity screening of drug candidates. The comparison of manual and automatic micronucleus analysis showed a high degree of concordance between the results obtained by the two techniques. For concentration series of cyclophosphamide (CP) and ethyl-methanesulphonate (EMS) as test compounds, the frequency of erroneously missed micronuclei through automatic scoring proved to be below 15% in comparison with manual scoring. Generally, false positive micronucleus decisions could be controlled easily by fast and simple relocation of the automatically detected patterns. The possibility to analyze 24 slides within 1 day by fully automatic overnight analysis and the high reproducibility of the results make automatic image processing a powerful tool for the in vitro micronucleus analysis.

Evaluation of the in vitro micronucleus test as an alternative to the in vitro chromosomal aberration assay: position of the GUM Working Group on the in vitro micronucleus test. Gesellschaft für Umwelt-Mutations-forschung.

In order to license a pharmaceutical or chemical, a compound has to be tested for several genotoxicity endpoints, including the induction of chromosomal aberrations in vitro. A working group within the GUM has evaluated published data on the in vitro micronucleus test with the aim of judging its suitability as a replacement for the in vitro chromosomal aberration test. After strict rejection criteria were applied, a database including 96 publications and 34 compounds was obtained. For 30 of these compounds, data on both tests were available. For 24 of the 30, concordant results in both test systems were obtained (80% correlation). The discordant results in 6 compounds can be explained by a known or suspected aneugenic potential of these compounds. Considering that cell types and test protocols were extremely heterogeneous, this correlation is rather encouraging. Comparison of the different protocols, and experience established within the working group yielded several recommendations for the routine use of the in vitro micronucleus test. Although many cell lines are suitable, those most often used in genotoxicity testing (e.g. CHL, CHO, V79, human lymphocytes, L5178Y mouse lymphoma cells) are recommended. Cytochalasin B may be used in the case of human lymphocytes; however, the possibility of its interaction with aneugenic test compounds should be considered. For continuously dividing cell lines, cytochalasin B is not recommended by the working group. Although, there seems to be flexibility in the choice of treatment and sampling times, the average generation time of the chosen cell line of choice should be taken into account when determining sampling time, and treatment of cells for at least one cell cycle duration is recommended. The use of appropriate cytotoxicity tests is strongly recommended. Although studies on some parameters of the test protocol may be useful, the introduction of the in vitro micronucleus test into genotoxicity testing and guidelines should not be delayed. Even in its present state, the in vitro micronucleus is a reliable genotoxicity test. Compared with the chromosomal aberration test, it detects aneugens more reliably, it is faster and easier to perform, and it has more statistical power and the possibility of automation.

Evaluation of a new procedure for the flow cytometric analysis of in vitro, chemically induced micronuclei in V79 cells.

Measurement of the frequency of micronuclei induced in cells by ionizing radiation or by chemical treatment is widely used to analyze cytogenetic damage. The microscopic scoring of micronuclei is a tedious and time-consuming procedure. Therefore, attempts have been made to automate micronuclei scoring by means of image analysis or flow cytometry. A new procedure for the flow cytometric analysis of chemically induced micronuclei in V79 Chinese hamster cells has been established in our laboratory. Debris was separated from micronuclei by means of a new gating procedure using area and width fluorescence of the stained suspension of micronuclei and nuclei. In order to test the sensitivity and specificity of this improved method of flow cytometric analysis, five well-known mutagenic compounds were tested. With the new technique, the frequency of micronuclei measured and analyzed corresponded well with results obtained by conventional microscopy. In addition, a large series of negative compounds, and weak, middle, and strong micronuclei inducers, were tested in order to establish criteria for discrimination between genotoxic and nongenotoxic compounds by flow cytometry. This new procedure for flow cytometric detection of micronuclei represents a quick, reliable, and relatively simple method for in vitro micronucleus testing.

Comparative evaluation of the in vitro micronucleus test and the in vitro chromosome aberration test: industrial experience.

Because of its rapidness, simplicity and potential for automation, the measurement of micronucleated cells in vivo is not only equivalent to the analysis of chromosome aberrations, but often even preferred within routine genotoxicity testing. In order to evaluate the correlation between the in vitro micronucleus assay (MNT) and the in vitro chromosome aberration test (CA), we collected data from four pharmaceutical companies obtained either in Chinese hamster cell lines (CHO-K5, CHO-K1, V79) or in human peripheral blood lymphocytes. Among the 57 compounds included in this comparison, 45 compounds gave rise to concordant results in both assays (26 compounds negative in both assays; 19 compounds positive in both assays). The high percentage of concordance, i.e. about 79% is very promising and can be even increased to about 88% by omitting the 3 aneugenic compounds and 2 compounds inducing endoreduplicated chromosomes which were found positive only in the in vitro MNT. The results are remarkable in particular considering that most of the compounds evaluated are 'standard' pharmaceutical compounds and thus are at most weak inducers of chromosome damage. Our comparison strongly supports that the in vitro micronucleus test is a suitable alternative to the in vitro chromosome aberration assay. Moreover, the MNT has the potential of not only detecting clastogens but additionally aneuploidy inducing chemicals.

[French language pharmaceutical publications in 1995. I. Assessment and analysis of French language pharmaceutical periodicals]. / La presse pharmaceutique francophone en 1995. I. Recensement et anlyse des Péruiduqes pharmaceutique francophones.

Periodicals constitute a vital source of information and an invaluable continuing education medium to pharmacists. The aim of the present study was to compile the list of the 83 pharmacy-related periodicals published in French as at 1995 with a view to establishing their main characteristics. The first part focuses on the definition of the concept of "French language pharmaceutical periodicals" selected for the survey needed a) to be in circulation in 1995 and published at least two times a year, b) published by pharmacists or non-pharmacists but must carry articles intended to help, inform, or retrain pharmacists irrespective of their specialties (community, hospital, biological or industrial pharmacy) as well as their closest collaborators (assistants, technicians, etc.) and c) published in French. The second section presents the methodology of the survey, which consisted essentially in consulting publisher and library catalogs and direct interviews with embassies and Pharmacy Associations in French-speaking countries. Questionnaires were also sent out to editors of periodicals and the information gathered was analysed by computer. The following section reproduces the results of the the survey: These were classified into three categories: scientific, professional, and continuing education periodicals. The last section concentrates on their readership (national, international), frequency, modalities of publication, date of creation, indexing in bibliographic databases.

Evaluation of the in vivo genotoxic potential of three carcinogenic aromatic amines using the Big Blue transgenic mouse mutation assay.

Three genotoxic mouse carcinogens, 4-chloro-o-phenylenediamine (4-C-o-PDA), 2-nitro-p-phenylenediamine (2-N-p-PDA), and 2,4-diaminotoluene (2,4-DAT), were tested in the Big Blue transgenic mouse mutation assay. Each experiment consisted of a vehicle control group with ten Big Blue C57BL/6 mice, five of either sex, and an equally sized group treated with a high dose of the test chemical. In addition, four animals were treated with the vehicle and six animals with the test compound for the measurement of bromodeoxyuridine (BrdU) incorporation to determine cellular proliferation. Prior to the mutagenicity experiments, the maximally tolerated dose of each compound was determined using nontransgenic C57BL/6 mice. Based on these results the doses used in the main study were 200 mg/kg/day for 4-C-o-PDA, 150 mg/kg/ day for 2-N-p-PDA, and 80 mg/kg/day for 2,4-DAT. Animals were treated for 10 days over a 2 week period and were killed 10 days after the ast treatment. In an additional experiment with 2,4-DAT, animals were killed 28 days after treatment. Since all three chemicals are liver carcinogens in the mouse, the DNA of the liver was analyzed using the standard procedures for the Big Blue assay. Hepatocyte proliferation was assessed by immunohistochemical detection of proliferating cell nuclear antigen (PCNA) and, in some studies, by measuring BrdU incorporation. 4-C-o-PDA and 2-N-p-PDA did not induce an increase in PCNA expression when measured 10 days after the last treatment. There was no increase in BrdU incorporation immediately after treatment with 4-C-o-PDA or with 2,4-DAT. However, 10 days after the last treatment with 2,4-DAT, a strong mitogenic effect was found with both techniques, i.e., in the PCNA and BrdU assays. 4-C-o-PDA, a liver carcinogen in both genders of mice, induced a small, statistically significant increase of the mutant frequencies in females. No increase was found in males. 2-N-p-PDA, which has been reported to induce liver tumors only in females, was found positive in males and was clearly negative in females. 2,4-DAT, a liver carcinogen in female mice, was positive in females and negative in males when the animals were killed 10 days after the last treatment. After an expression time of 28 days, 2,4-DAT induced a statistically significant increase in both sexes. The effect in females was marginally stronger than after 10 days' expression time and almost identical to the effect observed in males under these test conditions. In conclusion, the experiments showed that the Big Blue assay detects the genotoxicity of the three carcinogenic monocyclic aromatic amines tested. However, it seems that the sex specificity of the carcinogenic effects of these compounds is not reflected by the mutagenicity data in Big Blue mice.

[CD-ROM and pharmacy: a new means of accessing information. I. Characteristics and analysis of CD-ROMs of interest to pharmacy]. / CD-ROM et pharmacie: un nouveau moyen d'accès à l'information. I. Recensement et analyse des CD-ROM intéressant la pharmacie.

CD-ROMs represent a new data medium which is not used enough by health professionals. This survey aimed to list current CD-ROMs useful in pharmacy field with their main characteristics. Thus, databases and directories from health information providers were consulted. A database was then made in order to exploit data collected as well as possible. The following section detailed the results of the survey: 205 CD-ROMs were selected, 95% are bibliographic or full-text data, 94% use English language, 84% present international information, 82% allow us previous years information retrieval, 65% are updated at least every three months, 97% work on PC computers, 74% are provided by american companies.

Rapid detection and characterization of foot-and-mouth disease virus by restriction enzyme and nucleotide sequence analysis of PCR products.

Reverse transcription coupled with PCR was used for the detection of foot-and-mouth disease virus serotypes A, C, and O in organ extracts from experimentally infected cattle. Primers were selected from conserved sequences flanking the genome region coding for the major antigenic site of the capsid located in the C-terminal part of viral protein 1 (VP1). Because this region of the capsid is highly variable its coding sequence is considered to be the most appropriate for the characterization of virus isolates and, therefore, for the determination of the epidemiological relationships between viruses of the same serotype. For differentiation between serotypes and for detailed characterization of individual virus isolates restriction enzyme cleavage and nucleotide sequence analysis of the respective PCR products were carried out. In order to minimize the time required for sample preparation from clinical material, viral RNA was released from particles by heating the sample for 5 min at 90 degrees C. Finally, an air thermocycler was used, which allows performance of a PCR of 30 cycles in approximately 20 min. The results show that reverse transcription PCR followed by restriction enzyme analysis and/or nucleotide sequence analysis of the PCR products is useful for the rapid detection and differentiation of foot-and-mouth disease virus.

Survey of European pharmaceutical journals in circulation in 1993.

Journals represent a very important source of information to pharmacists. This survey aimed to list current European pharmaceutical journals with their main characteristics. First, we provide a working definition of "European pharmaceutical journal". Then, the methodology and our sources which included books, databases, and questionnaires are reported. The following section details the results of the survey: journal creation for the period 1930 to 1993, geographic distribution, characteristics of the different types of journals (scientific, professional, scientific and professional/educational journals), inclusion in international bibliographic indexes. Finally, the list of the European pharmaceutical journals with their main characteristics and the list of the independent European pharmaceutical journals are reported in Appendix.

Femoral neck abnormalities in spina bifida.

Femoral neck abnormalities in spina bifida can be of two types: Type A, consisting of widening of the physis and often associated with varus deformity, and Type B, characterized by marked narrowing of the femoral neck, resulting in a typical mushroom appearance. The Type A deformity is usually associated with an abduction contracture of the hip. This physeal lysis seems to be secondary to microtrauma sustained during persistent exercise done by the parents and therapist to overcome the abduction contracture. No treatment is required, even when varus deformity is present. These patients require an orthosis with a pelvic band for ambulation, and their mobility will not be affected by the deformity.