S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100001 La Revista Costarricense de Ciencias Médicas Salas Oviedo José Luis , 06 2001 2001 22 11 11
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100002 Efecto del dimetilsulfóxido en la respuesta quimioluminiscente y el consumo de oxígeno de neutrófilos humanos activados García Jorge ,Universidad de Costa Rica Escuela de Medicina Departamento de Bioquímica 06 2001 2001 22 17 32

El dimetil sulfóxido es un secuestrador inactivante del radical hidroxilo (.OH), e inhibe, de manera proporcional a la dosis, la quimioluminiscencia (QL) de luminol (QLU) y de lucigenina (QLC) en leucocitos polimorfonucleares neutrófilos (PMN) activados con estimulantes solubles y partículas de zimosán opsonizado (ZO). Los resultados indican que la inhibición de la QLU en respuesta al ionóforo de calcio A23187 puede deberse al secuestro de .OH por DMSO, mientras que la inhibición de la QLC sugiere que el DMSO afecta negativamente a la oxidasa de membrana de PMN. Ello se confirmó al observar que el DMSO inhibió el consumo de O2 en PMN activados con FMLP y ZO. Cuando el DMSO se añadió luego de la estimulación con FMLP y ZO, no hubo inhibición de la QLU, pero sí de la inducida por A23187. El lavado de PMN expuestos a DMSO causó un incremento en la QLU en respuesta a la estimulación con FMLP y ZO. Ello es congruente con la hipótesis de que el DMSO interfiere con la activación de las subunidades de membrana de la oxidasa por las unidades reguladoras citoplasmáticas. Estos resultados implican que el DMSO puede inhibir la QL en fagocitos tanto mediante secuestro de .OH como por interferencia con la producción de superóxido por la oxidasa de membrana.

Dimethylsulfoxide (DMSO), a hydroxyl radical scavenger, exerted a dose dependent inhibition on the luminol and lucigenin-enhanced chemiluminiscent responses of human neutrophils activated with soluble and particulate stimulants. DMSO inhibition of the luminol chemiluminescence induced by calcium ionophore A23187 was probably due to .OH scavenging, whereas inhibition of the lucigenin chemiluminiscence suggested DMSO negatively affects the NADPH-dependent membrane oxidase of neutrophils. In agreement with this, DMSO moderately inhibited O2 consumption in PMN suspensions stimulated with chemotactic peptide and opsonized zymosan. DMSO inhibition of chemotactic peptide and opsonized zymosan-induced luminol chemiluminescence was observed only when added before or in conjunction with stimulants, whereas A23187-induced chemiluminescence was inhibited by DMSO regardless of time of addition. Washing of DMSO-treated PMN resulted in increased luminol enhanced chemiluminescence in response to chemotactic peptide and opsonized zymosan. This is consistent with the idea that DMSO may be interfering with activation of the membrane subunits of the oxidase by translocation and docking of the cytoplasmic, regulatory subunits. These data imply that DMSO inhibits neutrophil chemiluminescence both by .OH scavenging and interfering with oxidase activation.

 Dimetil sulfóxido quimioluniniscencia luminol lucigenina neutrófilos Dimethylsulfoxide chemiluminiscent luminol lucigenin neutrophils Dujovny M. Rozario R. Kossovsky N. Diaz F.G. Segal R. Antiplatelet effect of dimethyl sulfoxide, barbiturates, and methyl prednisolone Ann NY Acad Sci 1983 411 Kharasch N. Thyagarajan B.S. Structural basis for the biological activities of dimethyl sulfoxide Ann NY Acad Sci 1983 411 Spruance S.L. McKeough M.B. Cardinal J.L. Dimethysulfoxide as a vehicle for topical antiviral chemotherapy Ann NY Acad Sci 1983 411 Wood D.C. Wood J. Pharmacological and biochemical consideration of dimethylsulfoxide Ann NY Acad Sci 1975 243 de la Torre J.C. Role of dimethylsulfoxide in prostaglandin-thromboxane and platelet systems after cerebral ischemia Ann NY Acad Sci 1983 411 Brown M. Desai M. Traber L.D. Herndon D.N. Traber D.L. Dimethylsulfoxide with heparin in the treatment of smoke inhalation injury J Burn Care Rehab 1988 9 Salim A.S. Oxygen-derived free radical scavengers prolong survival in gastric cancer Chemotherapy 1992 38 Salim A.S. Scavengers of oxygen-derived free radicals prolong survival in advanced colonic cancer A new approach Tumour Biol 1993 14 Smith G. Bertone A.L. Kaeding C. Simmons E.J. Apostoles S. Anti-inflammatory effectos of topically applied dimethyl sulfoxide gel on endotoxin-induced synovitis in horses Am J Vet Res 1998 59 Klein S.M. Cohen G. Cederbaum A.I. The interaction of hydroxyl radicals with dimethylsulfoxide produces formaldehyde FEBS Lett 1980 116 Repine J.E. Eaton J.W. Anders M.W. Hoidal J.R. Fox R.B. Generation of hydroxyl radical by enzymes, chemicals and human phagocytes in vitro. Detection with the anti-inflammatory agent, dimethylsulfoxide J Clin Invest 1979 64 Babbs C.F. Griffin D.W. Scatchard analysis of methane sulfinic acid production from dimethylsulfoxide: a method to quantify hydroxyl radical formation in physiologic systems Free Rad Biol Med 1989 6 Jacob S.W. Herschler R. Introductory remarks: dimethylsulfoxide after twenty years Ann NY Acad Sci 1983 411 Repine J.E. Fox R.B. Berger E. Effect of dimethylsulfoxide on the bactericidal function of polymorphonuclear leukocytes Ann NY Acad Sci 1983 411 Czuprynski C.J. Henson P.M. Campbell P.A. Effect of dimethylsulfoxide on the in vitro and in vivo bactericidal activity of human and mouse neutrophils and mononuclear phagocytes Inflammation 1984 8 Auer D.E. Ng. J.C. Seawright A.A. Superoxide production by stimulated equine polymorphonuclear leukocytes - inhibition by anti-inflammatory drugs J Vet Pharmacol Ther 1990 13 Beilke M.A. Collins-Lech C. Sohnle P.G. Effects of dimethylsulfoxide on the oxidative function of human neutrophils J Lab Clin Med 1987 110 Allen R.C. Phagocytic leukocyte oxygenation activities and chemiluminescence: a kinetic approach to analysis. Meth Enzymol 1986 133 Brestel E.P. Co-oxidation of luminol by hypochlorite and hydrogen peroxide: implications for neutrophil chemiluminescence Biochem Biophys Res Comm 1985 126 Dahlgren C. Aniansson H. Magnusson K.E. Pattern of Formylmethionyl-leucyl-phenylalanine-induced luminol and lucigenin-dependent chemiluminescence in human neutrophils Infect Immun 1985 47 Barber D.A. Do N.H. Tackett R.L. Capomacchia A.C. Nonsuperoxide lucigenin-enhanced chemiluminescence from phospholipid and human saphenous veins Free Radic Biol Med 1995 18 Repine J.E. Staer-Johansen K. Berger E. Hydroxyl radical scavengers produce similar decreases in the chemiluminescence responses and bactericidal activities of neutrophils Infect Immun 1984 43 Rao P.S. Luber, Jr. J.M. Millinowicz J. Lalezari P. Mueller H.S. Specificity of oxygen radical scavengers and assessment of free radical scavenger specificity efficiency using luminol enhanced chemiluminescence Biochem. Biophys. Res Comm 1988 150 Sadler K.L. Badwey J.A. Second messengers involved in superoxide production by neutrophils: function and metabolism Hematology/Oncology Clinics of North America 1988 2 Larfars G. Gyllenhammar H. Stimulus-dependent transduction mechanisms for nitric oxide release in human polymorphonuclear neutrophil leukocytes J Lab Clin Med 1998 132 Klausner R.D. Fishman M.C. Karnovsky M.J. Ionophore A23187 disrupts membrane stucture by modifying protein-lipid interactions Nature 1979 281 Borgeat P. Samuelsson B. Arachidonic acid metabolism in polymorphonuclear leukocytes: effect of ionophore A 23187 Proc Nat Acad Sci 1979 76 Singh D. Greenwald J.E. Bianchine J. Metz E.N. Sagone A.L. Evidence for the generation of hydroxyl radical during arachidonic acid metabolism by human platelets Amer J Hematol 1981 11 Van Dyke K. Matamoros M.C. Van Dyke C.J. Castranova V. Calcium ionophore stimulated chemiluminescence from human granulocytes: evidence that A23187 induced chemiluminescence originates from arachidonic acid metabolism Microchem J 1983 28 García J. La alfa hemolisina de Escherichia coli potencia el metabolismo oxidativo de neutrófilos humanos en respuesta al péptido quimiotáctico FMLP: comparación con el ionóforo de calcio A23187 Rev Cost Cienc Med 2000 21 O’Brien P.J. Hulett L.G. Hydroxyl radical involvement in the luminol chemiluminescence from the reaction of arachidonic acid with sheep vesicular gland microsomes Prostaglandins 1980 19 Radi R. Cosgrove T.P. Beckman J.S. Freeman B.A. Peroxynitrite-induced luminol chemiluminescence Biochem J 1993 290 Segal A.W. Shatwell K.P. The NADPH oxidase of phagocytic leukocytes Ann NY Acad Sci 1997 832
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100003 La exposición a ozono disminuye la adherencia y aumenta la permeabilidad de membrana de macrófagos alveolares de rata García Jorge ,Universidad de Costa Rica Escuela de Medicina Escuela de Medicina 06 2001 2001 22 33 44

El ozono es un gas generado por procesos fotoquímicos en áreas urbanas o industriales con niveles altos de contaminación, y su inhalación causa inflamación en vías respiratorias. Para estudiar el daño causado por el gas, macrófagos alveolares fueron obtenidos por medio de lavado pulmonar de ratas Sprague-Dawley, y expuestos a ozono (0,594 ppm) por un máximo de 60 minutos. Los efectos de la exposición se manifestaron como disminución de la adherencia celular a un sustrato de poliestireno, y un incremento en la permeabilidad de la membrana plasmática, manifiesto como aumento en la liberación específica de 51Cr, y como aumento en el calcio citoplásmico, evidenciado por medición espectrofluorométrica. Los resultados obtenidos apoyan la idea de que la membrana celular es el blanco principal de la acción del ozono. La elevación en el calcio citoplásmico podría mediar otras respuestas de los macrófagos al ozono, incluyendo la producción de eicosanoides y óxido nítrico, que acarrean una reducción concomitante en la capacidad fagocítica y la producción de superóxido.

Ozone gas is generated photochemically in areas with high levels of automotive or industrial emissions, and causes irritation and inflammation of the airways if inhaled. Rat alveolar macrophages were obtained by lung lavage from male Sprague Dawley rats and used as a model to assess ozone induced cell damage (0,594 ppm for up to 60 minutes). Ozone exposure caused loss of cell adherence to a polystyrene substrate and increased membrane permeability, as noted by increases in specific 51Cr release and citoplasmic calcium levels. The results indicate that the cell membrane is a target for ozone damage. Elevations of cytoplasmic calcium could mediate other macrophage responses to ozone, including eicosanoid and nitric oxide production, with concomitant decreases in phagocytic ability and superoxide production.

 ozono macrófagos alveolares permeabilidad de membrana ozone alveolar macrophages membrane permeability Ozone and other photochemical oxidants. Committee on Medical and Biologic Effects of Environmental Pollutants Ozone and other photochemical oxidants. Committee on Medical and Biologic Effects of Environmental Pollutants 1977 19 ^eWashington, D.C. Subcommittee on Ozone and Other Photochemical Oxidants. National Academy of Sciences Mustafa M.G. Biochemical basis of ozone toxicity Free Radical Biol Med 1990 9 Haagen-Smith A.J. Chemistry and Physiology of Los Angeles smog Ind Eng Chem 1952 44 Haagen-Smith A.J. Photochemistry and smog J Air Pollut Control Assoc 1963 13 Koren H.S. Devlin R.B. Graham D.E. Mann R. McGee M.P. Horstman D.H. Kozumbo W.J. Becker S. House D.E. McDonnell W.F. Bromberg P.A. Ozone-induced inflammation in the lower airways of human subjects Amer Rev Respir Dis 1989 139 Stephens R.J. Sloan M.F. Evans M.J. Freeman G. Early response of lung to low levels of ozone Am J Pathol 1974 73 Barry B.E. Miller F.J. Crapo J.D. Lee S.D. Mustafa M.G. Mehlman Alveolar epithelial injury caused by inhalation of 0.25 ppm ozone International Symposium on the biomedical effects of ozone and related photochemical oxidants International Symposium on the biomedical effects of ozone and related photochemical oxidants 1983 Princeton^eN.J. Princeton Scientific Publishers Brain J.D. Macrophage damage in relation to the pathogenesis of lung diseases Environ Health Perspect 1980 35 Becker S. Madden M.C. Newman S.L. Devlin R.B. Koren H.S. Modulation of human macrophage properties by ozone exposure in vitro Toxicol Appl Pharmacol 1991 110 Coffey M.J. Wheeler C.S. Gross K.B. Eschenbacher W.L. Sporn P.H. Peters-Golden M. Increased 5-lipoxygenase metabolism in the lungs of human subjects exposed to ozone Toxicology 1996 114 Scannell C. Chen L. Aris R.M. Tager I. Christina D. Ferrando R. Welch B. Kelly T. Balmes J.R. Greater ozone-induced inflammatory responses in subjects with asthma Am J Resp Crit Care Med 1996 154 Hamilton R.F. Li L. Eschenbacher W.L. Szweda L. Holian A. Potential involvement of 4-hydroxynonenal in the response of human lung cells to ozone Am J Physiol 1998 274 Banks M.A. Porter D.W. Martin W.G. Castranova V. Effects of in vitro ozone exposure on peroxidative damage, membrane leakage, and taurine content of rat alveolar macrophages Toxicol Appl Pharmacol 1990 105 Pasternak C.A. A novel role of Ca+2 and Zn+2: protection of cells against membrane damage Bioscience Reports 1988 6 Arsalane K. Gosset P. Vanhee D. Voisin C. Hamid Q. Tonnel A.B. Wallaert B. Ozone stimulates synthesis of inflammatory cytoquines by alveolar macrophages in vitro Am. J Respir Cell Mol Biol 1995 13 Dormans J.A. Rombout P.J. van Loveren H. Surface morphology and morphometry of rat alveolar macrophages after ozone exposure J Toxicol Environ Health 1990 31 Palmer R.M.J. Salmon J.A. Release of leukotriene B4 from human neutrophils and its relationship to degranulation induced by N-formyl-methionyl-leucyl-phenylalanine, serum-treated zymosan and the ionophore A23187 Immunology 1983 50 Canning B.J. Hmieleski R.R. Spannhake E.W. Jakab G.J. Ozone reduces murine alveolar and peritoneal macrophage phagocytosis: the role of prostanoids Am J Physiol 1991 261 Laskin D.L. Pendino K.J. Punjabi C.J. Rodríguez del Valle M. Laskin J.D. Pulmonary and hepatic effects of inhaled ozone in rats Environ Health Perspect 1994 102 ^s10 Radi R Cosgrove T.P. Beckman J.S. Freeman B.A. Peroxynitrite-induced luminol chemiluminescence Biochem J 1993 290 Ryer-Powder J.E. Amoruso M.A. Czerniecki B. Witz G. Goldstein B.D. Inhalation of ozone produces a decrease in superoxide anion radical production in mouse alveolar macrophages Amer Rev Resp Dis 1988 138 Hoffer E. Baum Y. Tabak A. Frevert C. Adhesion molecules of blood polymorphonuclear leukocytes and alveolar macrophages in rats: modulation by exposure to ozone Hum Exp Toxicol 1999 18
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100004 Determinación de ciclosporina sanguínea realizada en el Hospital San Juan de Dios, San José, Costa Rica Granados-Zúñiga Jorge ,Caja Costarricense de Seguro Social Hospital San Juan de Dios Laboratorio Clínico San José Costa Rica 06 2001 2001 22 45 50

Se determinó los niveles sanguíneos de ciclosporina y metabolitos (método de inmunoensayo de polarización de fluorescencia TDx, Abbott) y ciclosporina (método de inmunoensayo de polarización de fluorescencia AxSyM, Abbott) en 86 pacientes transplantados de riñón del Hospital San Juan de Dios. Los resultados fueron en promedio 3,26 veces mayores con el método de TDx que con el de AxSyM (amplitud=1,02-6,01, S.D.=0,99). Esto se puede atribuir a la reactividad cruzada del antisuero del TDx contra metabolitos de ciclosporina. La amplia variabilidad en las proporciones ciclosporina y metabolitos/ciclosporina (1,02-6,01) podría atribuirse a las diferencias interindividuales en la actividad de enzimas con citocromo P-450. Se sugiere que las diferencias en los métodos para determinar niveles de ciclosporina requerirían un ajuste en los márgenes terapéuticos de la droga.

Cyclosporine and metabolites (maded by FPIA Abbott TDx) and cyclosporine (maded by FPIA Abbott AxSyM) blood levels were assayed on 86 renal transplanted adult patients of any sex. Results were 3.26 times greater with TDx than with AxSyM (range=1,02-6,01, SD=0,99). That may be attributed to TDx antiserum cross-reactivity with cyclosporine metabolites. The wide variability in TDx/AxSyM rates (1,02-6,01) could be caused by interindividual differences on P-450 cytocrom-enzyme activities. It is suggested that differences in methods may require adjustments in therapeutic ranges.

 ciclosporina niveles sanguíneos de drogas transplante de riñón drug monitoring cyclosporine kidney transplant Kahan BD Cyclosporine N Eng J Med 1989 321 Belitsky P Steven D Johnston A Levy G Impact of absorption on efficacy and safety of cyclosporine therapy in transplant recipients Clin Pharmacokinet 2000 39 Kahan BD Dunn J Fitts C Reduced inter-and intrasubject variability in cyclosporine pharmacokinetics in renal transplant recipients treated with a microemulsion formulation in conjunction with fasting, low-fat meals, or high-fat meals Transplantation 1995 59 Lindholm A Kahan B Influence of cyclosporine pharmacokinetics, trough concentrations, and AUC monitoring on outcome after kidney transplantation Clin Pharmacol Ther 1993 54 Dumont RJ Ensom M Methods for clinical monitoring of cyclosporine in transplant patients Clin Pharmacokinet 2000 38 Herrera A Estudio retrospectivo transplante renal Hospital México, noviembre 1981-noviembre 1994 Estudio retrospectivo transplante renal Hospital México, noviembre 1981-noviembre 1994 1995 Vargas M Schosinsky K Aplicación del método gráfico de Levey-Jenning a los datos de un control de calidad externo en química clínica Rev Cost Cienc Med 1986 7 Vargas M. Vargas M Holst I Schosinsky K Brilla E Control de calidad interno Seguridad de calidad en Química Clínica Seguridad de calidad en Química Clínica 1989 San José Universidad de Costa Rica TDx immunosupressant drug assays-cyclosporine and metabolites whole blood TDx immunosupressant drug assays-cyclosporine and metabolites whole blood 1997 Illinois Abbott Laboratories Abbott AxSyM system cyclosporine Abbott AxSyM system cyclosporine 1999 Illinois Abbott Laboratories Wallemacq PE Alexandre K Evaluation of the new AxSyM cyclosporine assay: comparison with TDx monoclonal whole blood and emit cyclosporine assays Clin Chem 1999 45 Hebert MF Bioavailability of cyclosporine with concomitant revamping administration is markedly less than predicted by hepatic enzyme induction Clin Pharmacol Ther 1992 52 Turgeon DK Erythromycin breath test predicts oral clearance of cyclosporine in kidney transplant recipients Clin Pharmacol Ther 1992 52 Villeneuve JP Prediction of cyclosporine clearance in liver transplant recipients by the use of midazolam as a cytochrome P450A probe Clin Pharmacol Ther 2000 67 Ensom M Davis G Cropp C Ensom J Clinical pharmacokinetics in the 21st century Clin Pharmacokinet 1998 34 Holford N. The target concentration approach to clinical drug development Clin Pharmacokinet 1995 29 Morris R. Target concentration strategy for cyclosporin monitoring Clin Pharmacokinet 1997 32 Yee G McGuire T. Clin Pharmacokinet Phar ma co 19 Yee G McGuire T. Pharmacokinetic drug interactions with cyclosporin (Part II) Clin Pharmacokinet 1990 19
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100005 Un método rápido de reprocesamiento para microscopia electrónica de cortes histológicos en parafina Hernández-Chavarría Francisco Vargas-Montero Maribel Rivera Patricia Carranza Alfonso ,Universidad de Costa Rica Unidad de Microscopia Electrónica San José Costa Rica ,Universidad de Costa Rica Facultad de Microbiología San José Costa Rica ,CCSS Hospital Nacional de Niños San José Costa Rica 06 2001 2001 22 51 56

Se describe un método simple y rápido para el re-procesamiento de tejidos incluidos en parafina usados en microscopia de luz para poder ser observados con microscopia electrónica de transmisión (MET) y de rastreo (MER). Los tejidos incluidos en parafina se seccionaron, desparafinaron en tolueno y se expusieron a vapores de osmio mediante irradiación con microondas en un horno de microondas doméstico. Los tejidos fueron embebidos en resina epóxica, polimerizados y seccionados. El método de procesamiento requirió un tiempo relativamente corto (aproximadamente 30 minutos para MET y 15 para MER) y rinde una calidad razonable de la ultraestructura para propósitos diagnósticos.

A simple and rapid method is described for re-processing of light microscopy paraffin sections to observe they under transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The paraffin-embedded tissue is sectioned and deparaffinized in toluene; then exposed to osmium vapor under microwave irradiation using a domestic microwave oven. The tissues were embedded in epoxy resin, polymerized and ultrathin sectioned. The method requires a relatively short time (about 30 minutes for TEM and 15 for SEM), and produces a reasonable quality of the ultrastructure for diagnostic purposes.

 Tejidos embebidos en parafina microscopia electrónica de transmisión microscopia electrónica de rastreo Paraffin embedded tissues scanning electron microscopy transmission electron microscopy Afzelius BA Half a century of electron microscopy Ultrastr Pathol 1981 2 Hawkes PW The beginnings of electron microscopy The beginnings of electron microscopy 1985 Orlando Academic Press Inc Crang RFE Lkomparens KL Artifacts in biological electron microscopy Artifacts in biological electron microscopy 1988 New York Plenum Press Bozzola JJ Russell LD Electron microscopy: Principles and techniques for biologist Electron microscopy: Principles and techniques for biologist 1992 Boston Jones and Bartlett Publishers Bowers B Maser M Artifacts in fixation for transmission electron microscopy Artifacts in biological electron microscopy Artifacts in biological electron microscopy 1988 New York Plenum Press Newman GR Resin microscopy and on-section immuno-cytochemistry Resin microscopy and on-section immuno-cytochemistry 1993 Berlin Springer-Verlag Kok LP Boon ME Microwave cookbook for microscopists. Art and science of visualization Microwave cookbook for microscopists. Art and science of visualization 1992 3 Leiden Coulomb Press Hernández-Chavarría F Guillén R Microwave process for scanning electron microscopy Europ J Morphol 2000 38 Hernández-Chavarría F Vargas-Montero M Rapid staining of ultrathin sections using microwave irradiation with heat dissipation J Microsc 2001 202 Hernández F Kozuka Y Cambios post mortem en mucosa intestinal y tejido pulmonar de Mus musculus, observados al microscopio electrónico de barrido Rev Biol Trop 1981 29 Robinson DG Ehlers U Herken R Hermann B Mayer F Schürmann FW Methods of preparation for electron microscopy. An introduction for the biomedical sciences Methods of preparation for electron microscopy. An introduction for the biomedical sciences 1987 Berlin Spring-Verlag Ngai HK Chan KW Or SB Yau WL A rapid method for reprocessing paraffin sections for diagnostic electron microscopy J Pathol 1985 145 Mitin KS Pigolkin II A method for the re-embedding of tissues from paraffin blocks into epoxi resins for the study of stained semithinn sectiosn by electron microscopy Sud Med Ekspert 1997 40 Carranza A Rivera P Gutiérrez R Hernández-Chavarría F Endocarditis aguda estafilococcica fatal con periodontitis severa Rev Cost Cienc Med 2000 21 ^s3,4 Mather J Stanbridge CM Butler EB Method for the removal of selected cells from cytological smear preparations for transmission electron microscopy J Clin Pathol 1981 34 Bretscheider A Burns W Morrison A "Pop-off" technic. The ultrastructure of paraffin embedded sections Amer J Clin Pathol 1981 76 Widehn S Kindblom LG A rapid and simple method for electron microscopy of paraffin-embedded tissue Ultrastruct Pathol 1988 12
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100006 Cierre Asistido con Presión Negativa (VAC), en el tratamiento de esternotomía infectada: primer caso en Latinoamérica Elizondo Almeida Jorge Pucci Coronado Juan Soto- Pacheco Longino ,CCSS Hospital México Clínica de Heridas, Servicio y Cátedra Dermatología- Alergología Costa Rica ,CCSS Hospital México 2Servicio y Cátedra de Cirugía de Tórax y Cardiovascular Costa Rica 06 2001 2001 22 59 64

Una nueva técnica utilizando el cierre asistido con vacío o presión negativa (VAC, por sus siglas en inglés) se utilizó exitosamente en un paciente masculino de 57 años, con una herida de esternotomía dehiscente e infectada. Posterior a su desbridamiento y drenaje de absceso, el tratamiento regular con gasa iodoformada que utilizó por 10 días, fue sustituido con la aplicación de presión negativa con succión controlada durante 7 días. Esta novedosa técnica, para el cierre de heridas, utilizada por primera vez en Latinoamérica, en este tipo de lesión, hizo posible reducir el tiempo de cicatrización y los costos hospitalarios inherentes a este tipo de complicación infecciosa.

A new technique using vacuum assisted closure (VAC) was succesfully applied to a 57 year old, male patient, with a dehisced infected sternotomy wound. After debridment and abscess drainage, a standard treatment with iodorform gauze strip was applied during 10 days, then it was substituted with by the application of negative pressure by controlled suction through a porous derssing, during 7 days. This novel technique, the first time applied in Latinamerica in this kind of lesion, has made possible to reduce healing time and hospital costs.

 Cierre asistido con presión negativa costos de hospitalización Vacuum assisted closure hospital costs Raudat C.W Pagel J Woodhall D Early intervention and aggressive management of infected median sternotomy incision: a review of 2242 open-heart procedures Am Surg 1997 63 Tang A Ohri S Haw M Novel application of vacuum assisted closure technique to the treatment of sternotomy wound infection Eur J Orthop Surg Traumatol 2000 17 Obdeijn M Lange M Lichtendahl D Vacuum-assisted closure in the treatment of poststernotomy mediastinitis Ann Thorac Surg 1999 68 Morykwas M. J Argenta L.C Shelton-Brown E McGuirt W Vacuum-assisted closure: a new method for wound control and treatment: animal studies and basic foundation Ann Plast Surg 1997 38 Morykwas M.J Argenta Vacuum-assisted closure: a new method for wound control and treatment: Clinical Experience Ann Plast Surg 1997 38 Joseph E Hamori C.A Bergman S Swann N.F Anastasi G.W A prospective, randomized trial of vacuum-assisted closure versus standard therapy of chronic nonhealing wound Wounds 2000 12 ^s2 Meara J Guo L Smith J Vacuum-Assited Closure in the treatment of degloving injuries Ann Plast Surg 1999 42 Greer S.E Duthie E Cartolano B Techniques for applying subatmospheric pressre dressing to wounds in difficult regions of anatomy J WOCN 1999 26 Morykwas M.J David L.R Schneider A.M Whang C Jennings D.A Canty C.H Parker D White W.L Argenta L.C Use of subatmospheric pressure to prevent progression of Partial- Thickness Burns in a Swine Model J Burn Care Rehabil 1999 20 Argenta L.C Morykwas M.J Non-surgical modalities to enhance healing and care of soft issue wound J South Orthop Assoc 1997 6 ^s4 Rohrich R.J Lowe J.B Hackney F.L Bowman J.L Hobar P.C An algoritm for abdominal wall reconstruction Plast Reconstr Surg 2000 105 Genecov D.G Schneider A.M Morykwas M.J Parker D White W.L Argenta L.C A Controlled Subatmospheric Pressure Dressing Increases the Rate of Skin Graft Donor Site Reepithelialization Ann Plast Surg 1998 40 Schneider A.M Morykwas M.J Argenta L.C A New and Reliable Method of Securing Skin Grafts to the Difficult Recipient Bed Jour Plastic & Re-constructive Surgery 1998 102 ^s4 Rosser Ch Morykwas M Argenta L A new technique to manage perineal wounds Infections in Urology 2000 Mendez-Eastman S Negative Pressure Wound Therapy Plastic Surgical Nursing 1998 18 ^s1 Mc Callon S Knight C Valiulus J Vacuum-assisted closure versus saline-moistened gauze in the healing of postoperative diabetic foot wounds Ostomy/Wound Management 2000 46 ^s8 Wyoscki A.B Wound fluids and the pathogenesis of cronic wounds Journal of WOCN 1996 23 ^s6 Letsou G.V Rosales O Maltz S Vogt A Sumpio B.E Stimulation of Adenylate Cyclase Activity in Cultured Endothelial Cells Subjected to Cyclic Strech J Cardiovasc Surg 1990 31 Vandenburgh H.H Mechanical Forces and their second Messengers in Stimulating Cell Growth in Vitro Am J Physiol 1992 262 Sadoshima J Jahn L Toshiyuki T Kulik T.J Izumo S Molecular Characterization of the Strech-induced Adaptation of Cultured Cardiac Cells J Biol Chem 1992 267 ^s15 Philbeck T.E Whittington K.T Millsap M.H Briones R,B Wight D.G Schroeder W.J The Clinical and Cost Effectiveness of Externally Applied Negative Pressure Wound Therapy in the Treatment of Wounds in Home Healthcare Medicare Patients Ostomy/Wound Management 1999 45 ^s11
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100007 Satelismo plaquetario: primeros dos casos reportados en Costa Rica Cartín Walter Brenes María Oviedo Javier Segura Humberto ,Laboratorio Clínico Cartín y Ramos Alajuela Costa Rica ,Laboratorio Clínico Labisan Santa Ana Costa Rica 06 2001 2001 22 65 69

Menos de 100 casos de Satelitísmo Plaquetario se han descrito desde que en 1963, Field y MacLeod describieron por primera vez este fenómeno. Estos son los dos primeros casos del fenómeno descritos en la literatura Costarricense. El satelitismo plaquetario se observo en muestras sanguíneas con EDTA como anticoagulante y alrededor de los polimorfonucleares. A diferencia de otros casos descritos en la literatura la fagocitosis plaquetaria era evidente y no se observo pseudotrombocitopenia.

Field and MacLeod first reported the platelet satellitosis or satellitism in 1963 and to date is has been observed in no more than 100 cases. These are the first two cases reported in Costa Rica. This phenomenon occurred only in blood anticoagulated by EDTA and around polymorphonuclears. Pseudotrombocytopenia was not observed but platelet phagocytosis was evident.

 Satelitismo Plaquetario EDTA pseudotrombocitopenia fagocitosis plaquetaria Platelet satellitism EDTA pseudotrombocytopenia platelet phagocytosis Bizarro N Platelet Satellitosis to Polymorphonuclears: Cytochemical, lmmunological, and Ultrastructural Characterization of Eight Cases Am J Hematol 1991 36 Glassy E Glassy E Platelets Color Atlas of Hematology: An Illustrated Field Guide Based on Proficiency Testing Color Atlas of Hematology: An Illustrated Field Guide Based on Proficiency Testing 1998 North Field^eIllinois College of American Pathologist Bizarro N Goldschmeding R von Dem Borne A Platelet Satellitism Is Fc RIII (CDI 6) Receptor-Mediated Coag T Med 1994 Shahab N Evans M Platelet Satellitism N Engl J Med 1998 338 Morselli M Longo G Bonacorsi G Potenza L Emilia G Torelli G Anticoagulant C Mattock C Platelet pseudothrombocytopenia with platelet satellitism Haematologica 1999 84 Diacovo T Roth S Buccola J Baiton D Springer T Neutrophil Rolling, Arrest, and Transmigration Across Activated, Surface-Adherent Platelets Via Sequential Action of P-Selectin and the ß 32-Integrin CD11/ CD18 Blood 1996 88 Chirstopoulos C Mattock C Platelet Satellitism and ? granule proteins J CIin Pathol 1991 44 Evangelista V Manarini S Rotondo S Martelli N Polischuk R McGregor J PlateIet/Polymorphonuclear Leukocyte Interaction in Dynamic Conditions: Evidence of Adhesion Cascade and Cross Talk Between P-Selectin and the ? 32 lntegrin CD11b/CD18 Blood 1996 88 Govoni M Lodi G Menini C Ocurrence of Platelet Satellitism in Blood Donors Vox Sang 1991 61 Kabutomori O lwatani Y Koh T Amino N Platelet Satellitism Found by an Abnormal Leukocyte Cytogram With an Automated Cell Counter N Engl J Med 1998 338
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100008 Brucella: una bacteria virulenta carente de los factores de virulencia clásicos Freer Enrique Castro-Arce Rocío ,Universidad de Costa Rica Unidad de Microscopia Electrónica Costa Rica ,Universidad de Costa Rica Escuela de Medicina Departamento de Fisiología 06 2001 2001 22 73 82

La brucelosis es una enfermedad muy antigua conocida como fiebre de Malta. Fue descrita inicialmente en forma clínica por Marston en 1859 y Sir David Bruce aisló su agente causal en 1887. La brucelosis es una zoonosis de distribución cosmopolita que ha llegado a ser un serio problema económico y de salud pública en ciertos países. Los miembros del género Brucella son patógenos intracelulares facultativos que infectan una gran variedad de mamíferos. La bacteria se trasmite por ingestión o contacto con materiales contaminados provenientes de animales enfermos. A pesar de que algunos factores de virulencia han sido relacionados con Brucella, todavía no se conocen sus verdaderos factores de virulencia. Brucella no demuestra poseer factores de virulencia agresivos como exotoxinas, cápsula, plásmidos, fimbrias, flagelos o variación antigénica. Sin embargo, es muy virulenta y patogénica en sus huéspedes. Esta paradoja o controversia ha sido discutida por grupos de investigación que no aceptan la ausencia de verdaderos factores de virulencia y otros grupos que piensan que su baja actividad biológica in vitro e in vivo es per se un factor evolutivo de adaptación a la vida intracelular y como tal debe considerarse un factor de virulencia. Esta capacidad de adaptación esta probablemente relacionada con la composición de la membrana externa.

Brucellosis is an ancient disease known as Malta fever. Marston clinically described it for the first time in 1859 and its causative agent was isolated in 1887 by Sir David Bruce. Brucellosis is a zoonosis and is world wide distributed being a serious economic and public health problem in certain countries. Members of the genus Brucella are facultative intracelular pathogens, which infect and produce disease in a wide variety of mammals. Ingestion or contact with contaminated materials can infect humans. Although some factors have been implicated in the virulence of Brucella the exactly virulence mechanisms of this intracellular microorganism are not known yet. Brucella does not show aggressive virulence mechanisms such as exotoxins, anti-phagocitic capsules, plasmids, fimbria, flagella or antigenic variation. Even though it is highly pathogenic for preferred or accidental hosts. This paradigm has been discussed by groups who do not accept the absence of common virulent factors and groups that believe that the low in vitro and in vivo biological activities and silent capacity of Brucella to adapt to the intracellular environment is an evolutive virulence factor by itself. This capacity of intracellular adaptation is probably related to the outer membrane characteristic composition.

 Brucella factores de virulencia brucelosis membrana externa Brucella virulence factors brucellosis outer membrane Freer E Moreno E Moriyón I Pizarro-Cerda J Weintraub A Gorvel J-P Brucella-Salmonella lipopolysaccharide chimeras are less permeable to hydrophobic probes and more sensitive to cationic peptides and EDTA than their native Brucella sp. counterparts J Bacteriol 1996 178 Christie A.B. Brucellosis (undulant fever, Malta or Mediterranean fever) Infectious diseases: Epidemiology and Clinical Practice Infectious diseases: Epidemiology and Clinical Practice 1987 New York Churchill-Livinstone Crawford RP Huber JD Adams BS Nielsen K. Duncan J. R. Epidemiology and Surveinllance Animal Brucellosis Animal Brucellosis 1990 19 90 Boca Raton^eFlorida CRC Press Inc Mikolich DJ Boyce JM Bennet J.E. Brucella species Principles and practice of infectious diseases Principles and practice of infectious diseases 1990 NY Churchill Livingstone Inc Albert A. Selective toxicity Selective toxicity 1985 London Chapman and Hall Freer E Immunochemical characterisation of Brucella LPS and its relation to virulence Immunochemical characterisation of Brucella LPS and its relation to virulence Adams LG Adams LG. Advances in brucellosis research 1990 ^eTexas Texas A y M University Press Méresse S Steele-Mortimer O Moreno E Desjardins M Finlay B Gorvel JP Controlling the maturation of pathogen-containing vacuoles: a matter of life and death Nature Cell Biol 1999 1 Moore RA Bates NC Hancock R. W. Interaction of polycationic antibiotics with Pseudomonas aeruginosa lipopolysaccharide and lipid A studied by using dansyl-polymyxin. Antimicrob Agents Chemother 1986 29 Knight V. Beeson PB McDermott W Brucelosis Tratado de Medicina Interna de Cecil-Loeb Tratado de Medicina Interna de Cecil-Loeb 1977 Interamericana Fauci AS Braunwald E Isselbacher KJ Wilson JD Martin JB Kasper DL Hauser SL Longo DL Harrison Compendio de Medicina Interna Harrison Compendio de Medicina Interna 1999 14th Ed Interamericana Alton G.G. Jones L. M. Agnus R.D. Verger J.M. Techniques for the brucellosis laboratory Techniques for the brucellosis laboratory 1988 Paris Institut National de la Recherche Agronomique Holt JG Krieg NR Sneath PHA Staley JT Williams ST Bergey's Manual of Determinative Bacteriology Bergey's Manual of Determinative Bacteriology 1994 9th Baltimore Williams & Wilkins Moreno E. Pitt M. W. Jones L. M. Schurig G.G. Berman D.T. Purification and characterization of smooth and rough lipopolysaccharides from Brucella abortus J Bacteriol. 1979 138 Perera V.Y. Winter A.J. Ganem B. Evidence for covalent bonding of native-hapten protein complexes to smooth lipopolysaccharide of Brucella abortus FEMS Microbiol Lett 1984 21 Freer E. Production and characterization of monoclonal antibodies against Brucella lipid A and demonstration of the heterogeneity of Brucella lipid A by high-performance thin-layer chromatography Production and characterization of monoclonal antibodies against Brucella lipid A and demonstration of the heterogeneity of Brucella lipid A by high-performance thin-layer chromatography Gamazo C. Moriyón I. Release of outer membrane fragments by exponentially growing Brucella melitensis cells Infect Immun 1987 55 Moriyón I. Berman D.T. Effects of nonionic, ionic, and dipolar ionic detergents and EDTA on the Brucella cell envelope J Bacteriol 1982 Thiele O. W. Schwinn G. The free lipids of Brucella melitensis and Bordetella pertusis Eur J Biochem 1973 34 Freer E Rojas N. Weintraub A. Lindberg A. Moreno E. Heterogeneity of Brucella abortus lipopolysaccharides Res Microbiol 1995 146 Avhonen P Jansson E. Aho K. Marked cross-aglutination between Brucella and a subtype of Yersinia enterocilitica Acta Path Microbiol Scand 1969 75 Hurvel B Lindberg AA Serological cross-reactions between different Brucella species and Yersinia enterocolitica. Immunochemical studies on phenolwater extracted lipopolysaccharides from Brucella abortus and Yersinia enterocolitica type 9 Acta Path Microbio Scand 1973 81 Bundle DR Gidney MA Perry MB Duncan JR Cherwonogrodzky JW Serological confirmation of Brucella abortus and Yersinia enterocolitica O:9 O-antigens by monoclonal antibodies Infect Immun 1984 46 Caroff M Bundle DR Perry MB. Structure of the O-chain of the pheno-phase soluble cellular lipopolysaccharide of Yersinia enterocilitica. O:9 Eur J Bichem 1984 139 Corbel M.J. Parker M. T. Collier L. H. Brucella Topley’s and Wilson’s principles of bacteriology, virology and immunity Topley’s and Wilson’s principles of bacteriology, virology and immunity 1990 London Butler and Tanner Caroff M Bundle DR. Perry MB. Cherwonogrodzky JW. Duncan JR. Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3 Infect Immun 1984 46 Moreno E Jones LM. Berman DT. Immunochemical characterization of rough Brucella lipopolysaccharides Infect Immun 1984 43 Cherwonogrodzky JW Dubray G. Moreno E Mayer H Nielsen K. Duncan J. R. Antigens of Brucella Animal brucellosis Animal brucellosis 1990 Boca Ratón^eFlorida CRC Press Inc Moreno E Stackebrandt Dorsch M Wolters J Busch M Mayer H Brucella abortus 16S rRNA and lipid A reveal a phylogenetic relationship with members of the alpha-2 subdivision of the class Proteobacteria J. Bacteriol 1990 172 Moreno E. Plommet M. Evolution of Brucella Prevention of brucellosis is the mediterranean countries Prevention of brucellosis is the mediterranean countries 1992 Wageningen Pudoc Ewalt DR Payeur JB Martin BM Cummins DR Miller WG Characteristics of a Brucella species from a bottlenose dolphin (Tursiops truncatus) J. Vet. Diag. Invest. 1994 6 Enright M. Nielsen K. Duncan B. The pathogenesis and pathobiology of Brucella infection in domestic animals Animal brucellosis Animal brucellosis 1990 Boca Ratón^eFlorida CRC Press Inc. Shuterland SS Searson J Nielsen K. Duncan B. The immune response to Brucella abortus Animal brucellosis Animal brucellosis 1990 19 90 Boca Raton^eFlorida CRC Press Inc. Frenchick PJ Markam RJ F Cochrane A. Inhibition of phagosome-lysosome fusion in macrophages by soluble extracts of virulent Brucella abortus Am J Vet Res 1985 46 Canning P.C. Adams G. Phagocyte function in resistance to brucellosis Advances in brucellosis research Advances in brucellosis research 1990 College Station^eTexas Texas A & M University Press Detilleux PG Deyoe BL Cheville NF Penetration and intracellular growth of Brucella abortus in nonphagocytic cells in vitro Infect Immun 1990 58 Pizarro-Cerdá J Méresse S Parton RG van der Goot G Sola-Land A López-Goñi I Moreno E Gorvel JP Pathogenic Brucella abortus transits through the autophagic pathways and replicates in the endoplasmic reticulum of non profesional phagocytes Infect Immun 1998 66 Pizarro-Cerdá J Moreno E Gorvel JP Invasion and intracellular trafficking of Brucella abortus in nonphagocytic cells Microbes and Infection 20002 Alexander B Schnurrenberger PR Brown RR Numbers of Brucella abortus in the placenta, umbilicus and fetal fluid of two naturally infected cows Veterinary Record 1981 108 Moreno E. Genome evolution within the alpha Proteobacteria: why do some bacteria not posses plasmids and others exhibit more than one different chromosome? FEMS Microb Rev 1998 22 Bäumler AJ Heffron F Roth J. A. Bolin C. A. Brogden K., A. Minion F. C. Wannemuehler M. J. Microbial resistance to macrophage effector functions: Strategies for evading microbicidal mechanisms and scavenging nutrients within mononuclear phagocytes Virulence mechanisms of bacterial pathogens Virulence mechanisms of bacterial pathogens 1995 ^eWashimgton, D. C. ASM Press Ganz T Selsted ME Szklarek D Harwing SL Daher K Bainton DF Lehrer RI Defensins: natural peptide antibiotics of human neutrophils J Clin Invest 1985 76 Lehrer RI Ganz T Antimicrobial polypeptides of human neutrophils Blood 1990 76 Lehrer RI Ganz T Selste ME Defensins: endogenous antibiotic peptides of animal cells Cell 1991 64 Lehrer RI Lichtenstein AK Ganz T Defensins: antimicrobial and cytotoxic peptides of mammalian cells Annu Rev Immunol 1993 11 Goldstein J Hoffman T Frasch C Lizzio E F Beining PR Hochstein D Lee YL Angus RD Golding B Lipopolysaccharide (LPS) from Brucella abortus in less toxic than that from Escherichia coli, suggesting the possible use of B. abortus or LPS from B. abortus as carrier in vaccines Infect Immun 1992 60 Sawyer JG Martin NL Hancock REW Interaction of macrophage cationic proteins with the outer membrane of Pseudomonas aeruginosa Infect Immun 1988 56 Hindill RD Berman DT Antigens of Brucella abortus II: Toxicity for macrophages in culture I Infect Dis 1968 118 Leong D Diaz R Milner K Rudbach J Wilson JB Some estructural and biological properties of Brucella endotoxin Infect Immun 1970 1 Moreno E Berman DT Boettcher LA Biological activities of Brucella abortus lipopolysaccharides Infect Immun 1981 31 Moreno E. Características y actividades biológicas del lipopolisacárido de Brucella Adel. Microbiol. Enf. Infecc 1983 2 Freer Rassol O. Moreno E Jarstrand C Effect of Brucella abortus lipopolysaccharide on oxidative metabolism and lysozyme release by human neutrophils Infect Immun 1992 60
S0253-294820010001 Revista Costarricense de Ciencias Médicas Rev. costarric. cienc. méd 0253-2948 Editorial Nacional de Salud y Seguridad Social S0253-29482001000100009 Aseguramiento de la calidad analítica y norma ISO 17 025 en laboratorios clínicos y químicos Rodríguez-Benavides Gerarda Blanco-Sáenz Rigoberto ,Caja Costarricense Seguro Social Clínica Marcial Fallas Laboratorio Clínico San José Costa Rica ,Caja Costarricense Seguro Social Departamento Saneamiento Básico y Ambiental San José Costa Rica 06 2001 2001 22 83 97

Actualmente hay un gran interés en la estandarización de los sistemas de aseguramiento de calidad en los laboratorios, puesto que la aceptación y credibilidad de sus resultados depende de la identificación de las fuentes de variabilidad, de su control y de la documentación que así lo demuestre. Lo cual es resultado en buena medida de la globalización del mercado de bienes y servicios. En diciembre de 1999, se publicó la norma internacional ISO 17 025, que establece los requisitos administrativos y técnicos que debe cumplir un laboratorio de pruebas o calibración para obtener reconocimiento internacional, dentro del contexto de la Organización Mundial del Comercio. En este artículo se discuten los contenidos de la norma y su aplicación en laboratorios clínicos y químicos.

There is nowadays a strong concern for standardization of laboratory quality assurance systems, since results acceptability and confidence depend on identification of the sources of variability, their control and documentation, as a result to a greater extent of word commercial globalization. During December 1999, the International Standard ISO 17 025 was published, establishing organizational and technical requirements for test and calibration laboratories, in order to get international recognition, into the context of World Trade Organization. In this paper, such requirements are discussed for clinical and chemical laboratories.

 Aseguramiento de la Calidad Control de Calidad Norma ISO 17 025 Laboratorio Clínico Laboratorio Químico Analítico Quality Assurance Quality Control Standard ISO 17 025 Clinical Laboratory Chemical Analytical Laboratory Horwitz W Evaluation of Analytical Methods Used for Regulation of Foods and Drugs Analytical Chemistry 1982 54 Final Rule for Good Laboratory Practice Regulations under teh Federal Food Final Rule for Good Laboratory Practice Regulations under teh Federal Food 1978 Final Rule for Good Laboratory Practice Standards under teh federal Insecticide Final Rule for Good Laboratory Practice Standards under teh federal Insecticide 1983 Final Rule for Good Laboratory Practice Standards under the Toxic Substances Control Final Rule for Good Laboratory Practice Standards under the Toxic Substances Control 1983 Garne W Barge MS Good Laboratory Practices: An Agrochemical Perspective Good Laboratory Practices: An Agrochemical Perspective 1988 369 ^eWashington DC Good Laboratory Practices in the Testing of Chemicals: Final Report of the Group of Experts on Good Laboratory Practice Good Laboratory Practices in the Testing of Chemicals: Final Report of the Group of Experts on Good Laboratory Practice 1982 Paris Guía de la Ronda de Uruguay para la Comunidad Empresarial Guía de la Ronda de Uruguay para la Comunidad Empresarial 1996 Ginebra Aprobación del Acta Final en que se incorporan los resultados de la Ronda Uruguay de Negociaciones Comerciales Multilaterales La Gaceta 26 d e di 245 Guía de la Ronda de Uruguay para la Comunidad Empresarial Guía de la Ronda de Uruguay para la Comunidad Empresarial 1996 Ginebra Quality Assurance for Reserach and Development and Non-Routine Analysis Quality Assurance for Reserach and Development and Non-Routine Analysis 1998 First Ed Garfield Frederick M. Quality Assurance Principles for Analitycal Laboratories Quality Assurance Principles for Analitycal Laboratories 1996 Arlington^eV.A. AOAC International El-Nageh M.M. Heuck C.C. Appel W. Vandepitte J. Gibbs W.N. Engbaek K. Basics of Quality Assurance for Intermediate and Peripheral Laboratories Basics of Quality Assurance for Intermediate and Peripheral Laboratories 1992 Alexandria World Health Organization Sargent. M M. Revision of the VAM Principles Revision of the VAM Principles 1997 Middlesex Laboratory of The Government Chemist El-Nageh M.M. Heuck C.C. Appel W. Vandepitte J. Gibbs W.N. Engbaek K. Basics of Quality Assurance for Intermediate and Peripheral Laboratories Basics of Quality Assurance for Intermediate and Peripheral Laboratories 1992 Alexandria World Health Organization Westgard James O. Klee George G. Tietz Norbert W. Quality Assurance Fundamentals of Clinical Chemistry Fundamentals of Clinical Chemistry 1987 Third Philadelphia W. B. Saunders