Relationship Between the Dose Administered, Target Tissue Dose, and Toxicity Level After Acute Oral Exposure to Bifenthrin and Tefluthrin in Young Adult Rats.

Most pyrethroid insecticides (PYRs) share a similar primary target site in mammals. However, the potency estimates of the lethal and sublethal effects of these compounds differ up to 103-fold. The aim of this study was to evaluate the relationship between the dose administered, the target tissue dose, and the effect of 2 highly toxic PYRs, tefluthrin (TEF; 0.1-9 mg/kg) and bifenthrin (BIF; 0.5-12 mg/kg), by using the oral route, a corn oil vehicle (1 ml/kg) and subcutaneous temperature (Tsc) monitoring assays in adult rats. The Tsc was determined at 30-min intervals for 5 h (TEF) or 4.5 h (BIF) after dosing. Rats were sacrificed at 6 h after dosing, and BIF and TEF concentrations were determined in blood (Bd), liver (Lv), and cerebellum (Cb) by using a GC-ECD system. The minimal effective dose of BIF (3 mg/kg) affecting Tsc was similar to that found in prior studies using other testing paradigms. Regarding TEF, a very steep relationship between the dose administered and toxicity was observed, with a near-threshold to low-effective range for Tsc at 0.1-6 mg/kg, and a near lethal syndrome at ≥ 7.5 mg/kg. At 6-7.5 mg/kg TEF, the Cb/Bd and Cb/Lv concentration ratios were both > 1. Conversely, for BIF, the Cb concentration was barely over the Bd concentration and the Cb/Lv concentration ratio remained < 1. Our results and previous findings call for more comprehensive consideration to establish the relevance of the distribution into target tissues and the tissue dosimetry for health risks through the exposure to PYRs in humans.

Relationship between exposure, body burden and target tissue concentration after oral administration of a low-dose mixture of pyrethroid insecticides in young adult rats.

Pyrethroids (PYRs) are synthetic insecticides increasingly used in agricultural and household pest control. Little is known on how the toxicity of highly effective bolus doses of single compounds compares to more realistic scenarios of low-level exposure to PYR mixtures. In this study, we examined a quaternary mixture of two noncyano (tefluthrin, TEF; bifenthrin, BIF) and two cyano (α-cypermethrin, α-CPM; deltamethrin, DTM) PYRs in young adult rats. These compounds are mostly composed of PYR isomers ranking top ten in acute lethality in rats. Concurrently, we administered near-threshold levels of the four PYRs dissolved in corn oil by oral route. Six hours later blood was collected and the liver and cerebellum were dissected out to determine PYR concentrations in these tissues using Gas Chromatography with Electron Capture Detector (GC-ECD). The mixture caused mild-to-moderate changes in non-locomotor behaviors and subcutaneous body temperature (up to +1.2-1.5 °C increase at 2-4 h after dosing, respectively, compared to pre-dosing records). The most toxic PYRs BIF and TEF reached higher concentrations in the cerebellum than the cyano-compounds α-CPM and DTM. In addition, PYR concentrations in the cerebellum were correlated to single compound proportions in the dosing solution and changes in body temperature. Our results suggest that aggregate exposures resulting in a target tissue burden of ∼10-1 nmoles PYR/g may be toxicologically relevant, expanding the evidence on exposure-dose-effect relationships for PYRs, and serving to design convenient pharmacokinetic models for environmentally relevant exposures to PYR mixtures.

Hidroarsenicismo: mecanismos de acción asociados a la toxicidad del arsénico / Hydroarsenicism: Mechanisms of action related to arsenic toxicity

La exposición crónica al arsénico (As) inorgánico a través del agua de bebida da lugar al desarrollo de la enfermedad conocida como hidroarsenicismo. Esta enfermedad presenta sintomatología característica, sin embargo, para la mayoría de los efectos tóxicos que produce del As aún no se conoce en detalle el mecanismo de acción tóxica. Los mecanismos moleculares de acción del arsenito (unión a grupos sulfhidrilos) y del arseniato (sustitución del fosfato) están bien identificados, sin embargo, las consecuencias a nivel subcelular, celular, tisular y orgánico de esos mecanismos todavía presentan muchos huecos por llenar. A nivel subcelular y celular, la generación de especies reactivas de oxígeno (ERO) y de nitrógeno (ERN) son los mecanismos de acción tóxica del As más estudiados últimamente. Se los ha vinculado con la diferenciación y proliferación de queratinocitos, con la disfunción endotelial, con la resistencia a la insulina, con la inducción de peroxidación lipídica en hígado, de necrosis tubular renal y con cambios en la expresión del receptor estrogénico. Por último, la respuesta celular a proteínas no plegadas (como consecuencia del estrés del retículo endoplásmico) podría ser un mecanismo para explicar la afectación de la inmunidad humoral y la celular.

Chronic exposure to inorganic arsenic (As) through drinking water leads to the development of the disease known as hydroarsenicism. This disease presents characteristic symptomatology but the mechanisms underlying most of the toxic effects produced by As are not fully understand. The molecular mechanisms of action of arsenite (binding to sulfhydryl groups) and arsenate (phosphate substitution) are well identified, however, the consequences at the subcellular, cellular, tissue and organic levels of these mechanisms still have many gaps to fill. At the subcellular and cellular level, the generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) are the most studied mechanisms of toxic action. They have been linked to the differentiation and proliferation of keratinocytes, endothelial dysfunction, insulin resistance, induction of lipid peroxidation in the liver, renal tubular necrosis and changes in the expression of estrogen receptor. Finally, the cellular response to unfolded proteins (as a consequence of the stress of the endoplasmic reticulum) could be a mechanism to explain the affectation of humoral and cellular immunity.

Validación de un método para cuantificar ácido-11-nor-9-carboxi-delta 9-tetrahidrocannabinol en orina por cromatografía gaseosa-espectrometría de masas previa extracción en fase sólida / Validation of a method for quantify 11-nor-9-carboxy-delta 9-tetrahydrocannabinol in human urine using solid phase extraction and gas chromatography-mass spectrometry

El cannabis es una de las drogas ilegales más usadas a nivel mundial. Su consumo se relaciona con diferentes hechos en el ámbito forense, laboral, deportivo y clínico. Para su detección se utilizan métodos con diferentes fundamentos y alcances (inmunológicos, cromatográficos). En este trabajo se describe un método preciso, reproducible y validado, para la cuantificación del principal metabolito urinario del Δ9-tetrahidrocannabinol, el ácido-11-nor-9-carboxi- Δ 9-tetrahidrocannabinol (THC-COOH), por cromatografía gaseosa-espectrometría de masas (GC-MS). Se efectuó una extracción en fase sólida (SPE) a partir de la orina, previa hidrólisis alcalina. Se utilizó el análogo deuterado (THC-COOH D3) como estándar interno. El análisis por GC-MS se realizó en modo SIM. La curva de calibración fue lineal en el rango de trabajo (10-100 ng/ml, r > 0,999) y el límite de cuantificación fue de 10 ng/ml. La recuperación absoluta estuvo comprendida entre el 91,0 y 99,0 %. La precisión intra e inter ensayo fue de 1,06 a 1,26 y 3,59 a 9,80 %, respectivamente. El método fue aplicado a muestras reales, positivas por test inmunológico, resultando ser muy útil y fiable en el análisis de rutina de THC-COOH en orina humana con fines toxicológicos.

Cannabis is one of the most widely used illegal drug in the world. Its consumption is related to different forensic, work, sports and clinical events. In order to determinate the presence of cannabis, different methods with distinct fundamentals and scopes (immunoassay and chromatography) are applied. This report described an accurate, reproducible, and validated gas chromatography-mass spectrometry (GC-MS) method for the quantitation of 11-nor-9-carboxy- Δ9-tetrahydrocannabinol (THCCOOH), the major metabolite of Δ9-tetrahydrocannabinol in urine. A solid phase extraction (SPE), previous alkaline hydrolysis, was performed on the urine sample. Its deuterated analog (THC-COOH D3) was used as internal standard. The GC-MS analysis made by selected ion monitoring (SIM). Calibration curve was linear over the specified range (10 -100 ng/ml; r > 0.999) and limit of quantitation was 10 ng/ml. Absolute recoveries ranged from 91.0 to 99.0. Intra-assay and inter assay precision ranged from 1.06 to 1.26 and 3.59 to 9.80 %, respectively. The method has been applied to real samples, positive to immunological screening test, resulting to be very useful and reliable in routine analysis of THC-COOH in human urine for toxicological purposes.

Metabolitos del efavirenz como probable causa de falsos-positivos en test inmunológico para benzodiacepinas en orina / False-positive immunoassay results for urine benzodiacepines probably caused by efavirenz metabolites

En el tratamiento del síndrome de inmunodeficiencia adquirida (SIDA) una de las drogas antirretrovirales usadas es el efavirenz (EFV). Existe una asociación entre el consumo de drogas de abuso y la probabilidad de adquirir el SIDA, razón por la cual se solicita su investigación en orina. Como método de screening para detectar el consumo de estas drogas se utilizan habitualmente los ensayos inmunológicos. Una característica especial de estos métodos son las reacciones cruzadas que pueden presentarse con sustancias estructuralmente relacionadas dando origen a resultados falsos positivos.Al analizar 18 muestras de orina de pacientes con SIDA, se observó un 78% de resultados falsos positivos para benzodiacepinas (BZD) cuando fueron analizados mediante el ensayo inmunológico Triage® (Ascend Multi Immune Assay). El estudio confirmatorio por cromatografía gaseosa acoplada a espectrometría de masa (GC-MS) reveló la ausencia de BZD en todos los casos y el 100% de los resultados falsos positivos observados correspondieron a las muestras de los pacientes tratados con EFV. Con el propósito de dilucidar el origen de esta reacción cruzada fueron aislados el EFV y sus metabolitos de las muestras de orina, mediante extracción en fase sólida (SPE) y cromatografía en capa delgada de alta resolución (HPTLC), e identificados por cromatografía gaseosa- espectrometría de masa (GC-MS y GC-MS/MS/MS).Los resultados obtenidos en este estudio indicarían que los metabolitos del EFV (8-OH-EFV y/o 7-OH-EFV) y no el EFV podrían ser los responsables de la reacción cruzada observada en el ensayo inmunológico.

Efavirenz (EFV) is an antiretroviraldrug used in the acquired immunodeficiency syndrome (AIDS) treatment. Immunoassay techniques have been widely used for abuse drug screening test. The presence of structurally related substances in urine samples can interfere by cross reactionscausing false positive results. High percentage of false positive results (78%) for benzodiazepines (BDZ) had been established for 18 urine samples assayed by immunoassay test Triage® (Ascend Multi Immune Assay). Gas chromatography-mass spectrometry(GC-MS) evaluation was negative for BDZ for all cases. One hundred percent (100%) of the positive results came from patients treated with EFV. With the aim to determine the cause of this cross- reaction, EFV and its metabolites were isolated by solid phase extraction(SPE) and high performance thin-layer chromatography (HPTLC) and then, identified by gas chromatography mass spectrometry (GC-MS and GC-MS/MS/MS). GC-MS/MS/MS analysis showed that EFV metabolites (8-OH-EFV and/or 7-OH-EFV) could probably be responsible for the cross reaction observed in the immunologic assays.

Pharmacokinetic aspects of naloxone-precipitated morphine withdrawal in male and female prepubertal mice.

It has been shown that the expression of the morphine (MOR) withdrawal syndrome precipitated by naloxone (NAL) is more intense in male mice than in females, but the reasons for this phenomenon remain uncertain. The purpose of the present study was to evaluate whether this sexual dimorphism might be due to differences in MOR and/or NAL plasma levels after a chronic treatment with MOR. Prepubertal Swiss male and female mice were rendered dependent by intraperitoneal (i.p.) injection of MOR (2 mg/kg), twice daily for 9 days. On day 10 dependent mice received NAL (6 mg/kg, i.p.) 60 min after MOR injection. Blood samples were taken at different times in order to determine MOR and NAL plasma levels by gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC), respectively. Pharmacokinetic analysis showed no differences between male and female mice either for MOR or for NAL. In conclusion, although males and females respond differentially to NAL-precipitated withdrawal, this dimorphic behavior would not be influenced by a pharmacokinetic factor.

Residuos de plaguicidas organoclorados en leches infantiles y productos lácteos y su evaluación del riesgo / Risk assessment of organochlorine pesticides residues in infant milks and milk products

Se considera que la población infantil es el grupo mas sensible a los contaminantes provenientes de la ingesta de alimentos y debido a la alta relación consumo/ peso corporal. En vista de la problemática mundial de la contaminación ambiental por plaguicidas en Argentina no existe hasta el momento información sobre ingestas deplaguicidas y evaluación del riesgo en este grupo. Los alimentos infantiles (leches y productos lácteos) cubren una parte importante de los requerimientos nutricionales diarios para esta población pero al mismo tiempo puede contribuir con la exposición a contaminantes.El presente trabajo fue realizado a fin de investigar plaguicidas en alimentos especialmente desarrollados y recomendados para neonatos y niños. Se analizaron residuos de plaguicidas organoclorados en 50 muestras de leches maternizadas y en 51 muestrasde productos lácteos (postres y yugures) recogidas en el mercado local de Buenos Aires durante el período febrero de 2001 a diciembre de 2003. La investigación fue realizada mediante cromatografía gaseosa con detector de captura de electrones e inyección automática. Los resultados fueron expresados en ng/mL y se consideraron los siguientes grupos: €Aldrinas;€Clordano; €DDT; €Endosulfán; €HCH; €Heptacloro; Endrin; HCB y Lindano. Con los resultados obtenidos se calculó la Ingesta Diaria Estimada (IDE) en base al consumo de alimentos lácteos y el peso corporal. Los resultados fueron comparados con las IDAs recomendadas por el Codex Alimentarius. De las 101 muestras analizadas el 10 % no contiene ninguno de los plaguicidas investigados, 24,8% contiene al menos un pesticida, 19,7% contiene dos y el 2 % mostró evidencia de siete u ocho plaguicidas organoclorados de los diferentes grupos. De todos los grupos de pesticidas investigados las €Aldrinas y €DDT tuvieron porcentajes representativos (32%), y el€Heptachlor y €HCH las frecuencias mas elevadas, (57%) y (53%) respectivamente...

Children population is considered the most sensitive group to contaminants by food intake due to the high ratio food consumption/ body weight. Despite the worldwide problematic of pesticides environmental pollution in Argentine untilthis moment does not exist data about pesticide intake and risk assessment in this group. Baby food (milk and milk products) cover an important part of the daily nutritional requirements of this population but as the same time could contribute with the exposure to contaminants. The present work was aimed to investigate pesticides in products specially manufactured and recommended for neonates andinfants. Organochlorine pesticides residues were assessed on 50 milk samples and 51 milk products colleted at different local markets at Buenos Aires, between February 2001 and December 2003. The pesticides determination was performed by gas chromatography electron capture detection. The results were expressed in ng/mL and it was considered the following groups:€Aldrins; €Chlordane; €DDT; €Endosulfan; €HCH; €Heptachlor; Endrin; HCB and Lindane. With the result obtained the Estimate Daily Intake (EDI) was calculated on the basis of the daily consumption and the average body weight. The result obtained wascompared with the Acceptable Dialy Intake (ADI) recommended by the Codex Alimentarius. Among 101 samples analyzed, 10% did not contain pesticides residues, 24.8% contained at least one pesticide, 19.7% containedtwo and 2% showed evidence of seven and eight organochlorine pesticides. Among all pesticides groups investigated€Aldrins and €DDT had representative percentages (32%), and €Heptachlor (57%) and €HCH (53%) were the highest frequency groups...