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Background and purpose:  P‐glycoprotein (Pgp) efflux assays are widely used to identify Pgp substrates. The kidney cell lines Madin‐Darby canine kidney (MDCK)‐II and LLC‐PK1, transfected with human MDR1 (ABCB1) are used to provide recombinant models of drug transport. Endogenous transporters in these cells may contribute to the activities of recombinant transporters, so that drug transport in MDR1‐transfected cells is often corrected for the transport obtained in parental (wildtype) cells. However, expression of endogenous transporters may vary between transfected and wildtype cells, so that this correction may cause erroneous data. Here, we have measured the expression of endogenous efflux transporters in transfected and wildtype MDCK‐II or LLC cells and the consequences for Pgp‐mediated drug transport. Experimental approach:  Using quantitative real‐time RT‐PCR, we determined the expression of endogenous Mdr1 mRNA and other efflux transporters in wildtype and MDR1‐transfected MDCK‐II and LLC cells. Transcellular transport was measured with the test substrate vinblastine. Key results:  In MDR1‐transfected MDCK cells, expression of endogenous (canine) Mdr1 and Mrp2 (Abcc2) mRNA was markedly lower than in wildtype cells, whereas MDR1‐transfected LLC cells exhibited comparable Mdr1 but strikingly higher Mrp2 mRNA levels than wildtype cells. As a consequence, transport of vinblastine by human Pgp in efflux experiments was markedly underestimated when transport in MDR1‐transfected MDCK cells was corrected for transport obtained in wildtype cells. This problem did not occur in LLC cells. Conclusions and implications:  Differences in the expression of endogenous efflux transporters between transfected and wildtype MDCK cells provide a potential bias for in vitro studies on Pgp‐mediated drug transport.

Resistance to multiple antiepileptic drugs (AEDs) is a common problem in epilepsy, affecting at least 30% of patients. One prominent hypothesis to explain this resistance suggests an inadequate penetration or excess efflux of AEDs across the blood - brain barrier (BBB) as a result of overexpressed efflux transporters such as P-glycoprotein (Pgp), the encoded product of the multidrug resistance- 1 (MDR1, ABCB1) gene. Pgp and MDR1 are markedly increased in epileptogenic brain tissue of patients with AED-resistant partial epilepsy and following seizures in rodent models of partial epilepsy. In rodent models, AED-resistant rats exhibit higher Pgp levels than responsive animals; increased Pgp expression is associated with lower brain levels of AEDs; and, most importantly, co-administration of Pgp inhibitors reverses AED resistance. Thus, it is reasonable to conclude that Pgp plays a significant role in mediating resistance to AEDs in rodent models of epilepsy - however, whether this phenomenon extends to at least some human refractory epilepsy remains unclear, particularly because it is still a matter of debate which AEDs, if any, are transported by human Pgp. The difficulty in determining which AEDs are substrates of human Pgp is mainly a consequence of the fact that AEDs are highly permeable compounds, which are not easily identified as Pgp substrates in in vitro models of the BBB, such as monolayer (Transwell(®)) efflux assays. By using a modified assay (concentration equilibrium transport assay; CETA), which minimizes the influence of high transcellular permeability, two groups have recently demonstrated that several major AEDs are transported by human Pgp. Importantly, it was demonstrated in these studies that Pgp-mediated transport highly depends on the AED concentration and may not be identified if concentrations below or above the therapeutic range are used. In addition to the efflux transporters, seizure-induced alterations in BBB integrity and activity of drug metabolizing enzymes (CYPs) affect the brain uptake of AEDs. For translating these findings to the clinical arena, in vivo imaging studies using positron emission tomography (PET) with (11)C-labelled AEDs in epileptic patients are under way.

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Thus, these data indicate that brain levels of topiramate may be affected by overexpression of Pgp as determined in patients with intractable epilepsy.

Medroxiprogesterona acetato para la elaboración de dispositivos intravaginales caseros usados en la sincronización del estro en ovinos de pelo: Evaluación de su eficacia, sus efectos secundarios y comparación con un dispositivo comercial en hatos de Costa Rica. Resumen El objetivo del trabajo fue determinar la eficacia farmacológica y los efectos secundarios de un dispositivo intravaginal de fabricación casera elaborado con acetato de medroxiprogestrona para la sincronización del estro en ovinos de pelo. Se incluyeron un total de 78 hembras adultas, multíparas y ciclando en un estudio de dos etapas. En la primera etapa se utilizaron 18 animales con el objetivo de analizar la eficacia y los efectos secundarios del dispositivo de elaboración casera. En la segunda etapa se utilizaron 60 hembras divididas aleatoriamente en dos grupos: (G1) dispositivo casero (n-30), (G2) dispositivo comercial (n-30). El fin de esta etapa fue comparar la tasa de sincronización de celos y la tasa de concepción por monta natural. Para la sincronización se colocó el dispositivo con 60mg de acetato de medroxiprogesterona durante 12 días, con inyección de 200 UI de eCG y 5 mg de dinoprost trometamina al retirar del dispositivo. Se evaluaron andrológicamente 13 machos, de los cuales 4 fueron descartados por ser insatisfactorios y se utilizaron 8 machos satisfactorios durante ambas fases del estudio. En ambas etapas la presentación de celo fue nula durante la presencia del dispositivo y 100% presentaron celo 48 horas después de su retiro. Los efectos adversos en la primera etapa fueron: Vaginitis leve al día 12 en todas las hembras, 3 presentaron adherencias, 2 hembras con aumento de fibrinógeno el día 12 y una disminución general de los leucocitos. En la segunda etapa no hubo diferencias en la sincronización entre los grupos, la tasa de concepción fue de 80,0% y 76,7% en G1 y G2, respectivamente. Basado en los resultados de este trabajo, el dispositivo intravaginal de fabricación casera es efectivo para la sincronización del estro en ovinos de pelo y para la obtención de altas tasas de preñez por monta natural.Palabras clave: Sincronización del estro, ovinos, medroxiprogesterona acetato. AbstractThe goal of our study was to determine the pharmacological efficacy and adverse effects of a homemade intravaginal device containing medroxyprogesterone acetate for estrus synchronization in hair ewes. A total of