Patch clamp electrophysiology serves as a powerful method for studying changes in plasma membrane ion conductance induced by externally applied high-intensity nanosecond electric pulses (NEPs). This paper describes an enhanced monitoring technique that minimizes the length of time between pulse exposure and data recording in a patch-clamped excitable cell. Whole-cell membrane currents were continuously recorded up to 11 ms before and resumed 8 ms after delivery of a 5-ns, 6 MV/m pulse by a pair of tungsten rod electrodes to a patched adrenal chromaffin cell maintained at a holding potential of -70 mV. This timing was achieved by two sets of relay switches. One set was used to disconnect the patch pipette electrode from the pre-amplifier and connect it to a battery to maintain membrane potential at -70 mV, and also to disconnect the reference electrode from the amplifier. The other set was used to disconnect the electrodes from the pulse generator until the time of NEP/sham exposure. The sequence and timing of both sets of relays were computer-controlled. Using this procedure, we observed that a 5-ns pulse induced an instantaneous inward current that decayed exponentially over the course of several minutes, that a second pulse induced a similar response, and that the current was carried, at least in part, by Na. This approach for characterizing ion conductance changes in an excitable cell in response to NEPs will yield information essential for assessing the potential use of NEP stimulation for therapeutic applications.
The mechanism of TMI‐induced crosslinking was studied using model reactions in a mixture of TMI, water and acetic acid. The hydrolysis of TMI was slow at room temperature, but could be enhanced by the incorporation of carboxylic acid groups. The formation of amide linkages via condensation between TMI and carboxylic acid groups did occur at room temperature, but was very slow. The crosslinking of a fully crosslinked latex film was also studied using a new type of latex. The degree of crosslinking of the core‐crosslinked latex films was lower than theoretical values and the mechanical properties were poor, which was ascribed to insufficient interfacial crosslinking.magnified image
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