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.

Daily physicochemical, microbiological and ecotoxicological fluctuations of a hospital effluent according to technical and care activities.

The problem of hospital effluents falls into the framework of hazardous substances due to the specific substances used and discharged for the most part into urban drainage networks without prior treatment. This in-depth study has led to greater understanding of the effluents discharged by hospitals. The experimental program implemented consisted in carrying out parallel sampling of the effluents of one hospital: a 24 h-average sample and 5 periodic samples corresponding to fractions of times and hospital activities. The samples were characterized by physicochemical, microbiological and ecotoxicological analyses. The results highlight that the effluents contained very little bacterial flora and a moderate organic pollution. However, a numerous of specific pollutants were detected: AOX, glutaraldehyde, free chlorine, detergents, Freon 113 as well as alcohols, acetone, formaldehyde, acetaldehyde, ammonium, phenols and several metals. The battery of bioassays showed that the effluents had a high level of ecotoxicity partly linked to particles in suspension and, that pollution fluctuated greatly during the day in connection with hospital activities. Finally, the PNEC values compared to the concentrations of pollutants dosed in the effluents highlighted that their toxicity was mainly due to several major pollutants, in particular free chlorine. Some hypotheses require additional experiments to be carried out. They concern: reactions of fermentations likely to occur in the drainage network and to form secondary toxic compounds, retention of chlorine by particles and physicochemical characterization of suspended solids.

[Method selected for the determination of creatinine in plasma or serum. Choice of optimal conditions of measurement]. / Technique sélectionnée pour le dosage de la créatinine dans le plasma ou le sérum, Choix des conditions optimales de mesure.

The method selected by the SFBC (Société française de biologie clinique) is derived from the colorimetric reaction of creatinine with alkaline picrate, measured kinetically, without any pretreatment step. The key parameters of the reaction determining the quality of the results are studied, with special regard to samples including known interferents. The aims of the study were to gain an optimal analytical sensitivity and to reduce main interferences (acetoacetate, bilirubine, glucose, protein) which plague the Jaffé reaction, through a comprehensive study of the reagents, of their concentrations and of the analytical procedures. The selected concentrations (in the test) are: 150 mmol/L sodium hydroxide, 10 mmol/L picric acid and 2 g/L sodium dodecyl sulfate. Ten millilitres of a BRIJ solution (30% volvol) are added to the reagent. The operating procedures are as follow: sample ratio 0.07 to 0.08; wavelength 505 to 510 nm; temperature 37 degrees C; incubation of the specimen with the alkaline reagent 5 mn (at least), before starting the reaction with picric acid. A seric calibrator is recommended. The first measurement is taken 20 to 40 s after starting the reaction. Total measurement time is 120 to 150 seconds.

[Validation and study of the transferability of the selected technique by SFBC for the determination of creatinine in plasma]. / Validation et étude de la transférabilité de la technique sélectionnée par la SFBC pour le dosage de la créatinine dans le plasma.

A selected method for the determination of creatinine in plasma, using the reaction with alkaline picrate without prior pretreatment has been proposed by the Commission 'Validation de techniques' in the SFBC (Société Française de biologie clinique). The transferability step was conducted in seven laboratories, equipped with different automatic analyzers, using analytical procedures derived from the recommended method. Its goal was to test whether the original analytical performances could be maintained and consistent results obtained. The validation step was designed to evaluate the linearity limits of the analytical range, the detection limit, to assess accuracy as compared to a high performance liquid chromatography and to investigate the effect of the main interferents. Linearity limits are 15 and 2000 mumol/L. The detection limit is 3 to 8 mumol/L according to the analytical systems. The selected method can fulfil the set imprecision goals: intralaboratory CV minus than 2% (within-run), minus than 4% (run-to-run), interlaboratory CV minus than 5% (for 100 mumol/L creatinine). Inaccuracy evaluated for the chosen control sera is 1 to 15% as compared to the chromatographic method, according to the sera and to the analytical systems. The results obtained with the selected method are more consistent with the HPLC than are those obtained with an alkaline picrate method without SDS or with an enzymatic method. No interference could be demonstrated for acetoacetate (up to 8 mmol/L), hemoglobin (up to 210 mumol/L), unconjugated bilirubin (up to 250 mumol/L), glucose (up to 30 mmol/L), IgG (up to 45 g/L), albumin (up to 60 g/L). The effect of cephalosporins depends on the molecule. The reagents are stable for at least 6 months when stored in closed vials at +20 degrees C. The alkaline reagent is stable 30 days at +4 degrees C. Reference limits (0.025 and 0.975 fractiles) have been established for healthy adults. They are respectively 73 to 126 mumol/L for men and 59 to 100 mumol/L for females.

Liquid-phase reactions started by rehydrating lyophilized reagents in a centrifugal analyzer.

In the disposable rotor of the SAM microcentrifugal analyzer, various lyophilized reagents are predistributed in 24 33-microL cuvets, for determination of multiple analytes in one specimen (e.g., for patient profiles). We evaluated a prototype of this system, which can be used at 25, 30, or 37 degrees C; absorbance readings at 340, 405, and 500 nm varied linearly up to 2.0 A. Starting the reactions by rehydrating the reagents with diluted serum is adequate because absorbance readings do not begin until 180 s after initiating the rehydration. Analytical performances of kinetic determinations at 30 degrees C showed good accuracy and correlation with other methods for creatine kinase, amylase, aspartate aminotransferase, and gamma-glutamyltransferase. Kinetic determinations for urea, and equilibrium determinations with blank corrections for glucose, cholesterol, and triglycerides gave excellent results for glucose and correct results for the other analytes. This compact analyzer combines the analytical performances of a centrifugal analyzer with the practicability of instruments having predistributed reagents.

Different fates of hepatitis B virus markers during plasma fractionation: a clue to the infectivity of blood derivatives.

Hepatitis B surface antigen (HBs Ag) and associated particles, e antigen (e Ag) and DNA polymerase are unevenly distributed during Cohn's cold ethanol fractionation of plasmas positive for these markers of the hepatitis B virus (HBV). Most of the e Ag, Dane particles and DNA polymerase are retained in fraction III whereas the bulk of HBs Ag is recovered in fraction IV where only 22 nm spheres and short filaments are still identified. These results suggest that differences in quantitative distribution of HB virions together with alteration of infectious particles during the fractionation process may in addition to heat inactivation account for the relative hepatitis risk of the various plasma derivatives.

[Comparative study of three technics of inverse passive hemagglutination in the search for a surface antigen of the hepatitis b virus (Ag HBs) (author's transl)]. / Etude comparative de trois techniques d'hémagglutination passive inverse pour la recherche de l'antigène de surface du virus de l'hépatite B (Ag HBs).

The following three different versions of reverse passive hemagglutination (RPHA) were evaluated for the detection of HBs antigen: Auscell I - Abbott, WH HBs - Wellcome and the hepanosticon - Organon technique and results obtained were compared with those obtained by the radio immuno assay (Ausria II of Abbott). In 493 sera studied, up to 16,8% were found positive by RPHA as compared to 17,2% positives by RIA. The percentage of false positives by the different methods varied from 4,9 to 7,3. Confirmatory tests, either absorption or neutralization, are necessary to ascertain accuracy of positive results in each of the 3 RPHA methods. The high quality of the Auscell and WH HBs confirmative test allows their sole use although they are slightly less sensitive than the RIA. We would recommand use of the Hepanosticon test whenever positive sera can be confirmed by RIA.

[Detection and prognostic significance of Ag "e" and its antibody in the course of HB virus infections[]. / Détection et signification pronostique de l'Ag "e" et de son anticorps au cours des infections à virus HB

A new antigen antibody system the e Ag and Ab has been investigated by immunodiffusion and counterelectrophoresis in the serum of 509 subjects. Those included 242 patients with polyarteritis, acute chronic or fulminant hepatitis; 85 hemodialysis HBs Ag carriers and 182 asymptomatic HBs Ag carrier blood donors. Neither e Ag nor anti- e were detected in any of the non hepatitis B virus associated cases. Counterelectrophoresis was found to be more sensitive than immunodiffusion and detected either e Ag or anti- e in 24 p. cent more cases. e Ag was found among HBs Ag positive patients with polyarteritis, chronic hepatitis or under going hemodialysis. Anti- e was observed in 28 p. cent to 66 p. cent of asymptomatic HBs Ag carriers. The study of the nature and prognostic significance of e Ag and anti- e appear of major importance in the understanding and follow up of HB virus infections.

Hepatitis B antigen (HBSAg) and/or antibodies (anti-HBS and anti-HBC) in fulminant hepatitis: pathogenic and prognostic significance.

Hepatitis B surface antigen (HBSAg) and antibodies to both the surface and core antigens of the hepatitis B virus (anti-HBS and anti-HBC) have been studied in 64 consecutive cases of fulminant hepatitis. HBSAg was detected by counterelectrophoresis in 23 (35-9%) but by radioimmunoassay in 38 (59-3%). Anti-HBS was detected by passive haemagglutination in 26 (40-6%), coexisting HBSAg and anti-HBS were found in 16 cases (25%). Using an indirect immunofluorescence technique, anti-HBC was found in all of the cases in whom either HBSAg or anti-HBS was present. The highest survival rate was observed in patients with no evidence of HBV infection (31-3%) and was lowest in those who had both HBSAg and anti-HBS detected simultaneously (6-2%). The prognosis of those who exhibited anti-HBS only was no better than those with HBSAg alone. In a further case, transient interruption of the asymptomatic chronic HBSAg carrier state with seroconversion to anti-HBS was associated with the development of a fulminant hepatitis syndrome. The results suggest that an unusually strong and rapid immune clearance of HBSAg may be involved in the pathogenesis of fulminant hepatitis.