Optimal salt concentration of vehicle for plasmid DNA enhances gene transfer mediated by electroporation

In vivo electroporation has emerged as a leading technology for developing nonviral gene therapies, and the various technical parameters governing electroporation efficiency have been optimized by both theoretical and experimental analysis. However, most electroporation parameters focused on the electric conditions and the preferred vehicle for plasmid DNA injections has been normal saline. We hypothesized that salts in vehicle for plasmid DNA must affect the efficiency of DNA transfer because cations would alter ionic atmosphere, ionic strength, and conductivity of their medium. Here, we show that half saline (71 mM) is an optimal vehicle for in vivo electroporation of naked DNA in skeletal muscle. With various salt concentrations, two reporter genes, luciferase and beta-galactosidase were injected intramuscularly under our optimal electric condition (125 V/cm, 4 pulses x 2 times, 50 ms, 1 Hz). Exact salt concentrations of DNA vehicle were measured by the inductively coupled plasma-atomic emission spectrometer (ICP-AES) and the conductivity change in the tissue induced by the salt in the medium was measured by Low-Frequency (LF) Impedance Analyzer. Luciferase expression in-creased as cation concentration of vehicle dec-reased and this result can be visualized by X-Gal staining. However, at lower salt concentration, transfection efficiency was diminished because the hypoosmotic stress and electrical injury by low conductivity induced myofiber damage. At optimal salt concentration (71 mM), we observed a 3-fold average increase in luciferase expression in comparison with the normal saline condition (p < 0.01). These results provide a valuable experimental parameter for in vivo gene therapy mediated by electroporation.

Optimal salt concentration of vehicle for plasmid DNA enhances gene transfer mediated by electroporation

In vivo electroporation has emerged as a leading technology for developing nonviral gene therapies, and the various technical parameters governing electroporation efficiency have been optimized by both theoretical and experimental analysis. However, most electroporation parameters focused on the electric conditions and the preferred vehicle for plasmid DNA injections has been normal saline. We hypothesized that salts in vehicle for plasmid DNA must affect the efficiency of DNA transfer because cations would alter ionic atmosphere, ionic strength, and conductivity of their medium. Here, we show that half saline (71 mM) is an optimal vehicle for in vivo electroporation of naked DNA in skeletal muscle. With various salt concentrations, two reporter genes, luciferase and beta-galactosidase were injected intramuscularly under our optimal electric condition (125 V/cm, 4 pulses x 2 times, 50 ms, 1 Hz). Exact salt concentrations of DNA vehicle were measured by the inductively coupled plasma-atomic emission spectrometer (ICP-AES) and the conductivity change in the tissue induced by the salt in the medium was measured by Low-Frequency (LF) Impedance Analyzer. Luciferase expression in-creased as cation concentration of vehicle dec-reased and this result can be visualized by X-Gal staining. However, at lower salt concentration, transfection efficiency was diminished because the hypoosmotic stress and electrical injury by low conductivity induced myofiber damage. At optimal salt concentration (71 mM), we observed a 3-fold average increase in luciferase expression in comparison with the normal saline condition (p < 0.01). These results provide a valuable experimental parameter for in vivo gene therapy mediated by electroporation.

Medicación preanestésica por vía oral con midazolam y ketamina en jugo de manzana, para pacientes pediátricos sometidos a cirugía ambulatoria / Preanesthetic medication with midazolam or ketamine in apple juice for outpatient surgery in childrens

La medicación preanestésica es de gran utilidad para los niños sometidos a cirugía electiva ambulatoria, se administra fácilmente y es bien aceptada por vía oral. Se administraron dosis de midazolam y ketamina por vía oral de 0.75 mg/kg y 6 mg/kg respectivamente utilizando como vehículo jugo de manzna, en dos grupos de pacientes de 20 cada uno, en niños de 1-10 años de edad. Se observó el grado de sedación, calidad de la separación familiar y facilidad para la aplicación de venopunción a los 15 y 30 minutos después de su administración, grado de amnesia y tiempo de recuperación. El inicio de acción y calidad de sedación resultaron con p< 0.05 a favor del midazolam, el resto de las observaciones registradas fueron similares en ambos grupos

Behavior effects of adrenaline and noradrenaline injected into the lateral ventricles of the pigeon (Columba Livia)

The behavioral effects of intravrnticular 1-micronl injections of adrenaline and noradrenaline (both in a concentration of 30 nmol/micronl) were wxamined in pigeons bearing cannule chronically implanted into the lateral ventricles. Injections of either catcholoamine evoked immediate and intense bouts of feeding behavior, followed by long-lasting incrases in sleep duration (50-90% higher than vehicle-treated subjects) in pigeons given free access to food during the observation period. Pigeons treated with adrenaline or vehicle only, and placed in a cage without the feeder set (food-deprived durngi the observation period), exhibited late increases in exploratory and preening behaviors, and less sleep than controls (vehicle-treated pigeons with free access to food). These data suggest that post-prandial sleep in this situation may represent a by-product of feeding-related processes evoked by both catecholamines