The refolding kinetics of alpha-lactalbumin at different concentrations of guanidine hydrochloride have been investigated by means of kinetic circular dichroism and stopped-flow absorption measurements. The refolding reaction consists of at least two stages, the instantaneous accumulation of the transient intermediate that has peptide secondary structure and the subsequent slow process associated with formation of tertiary structure. The transient intermediate is compared with the well-characterized equilibrium intermediate observed during the denaturant-induced unfolding. Stabilities of the secondary structures against the denaturant, affinities for Ca2+, and tryptophan absorption properties of the transient and equilibrium intermediates were investigated. In all of these respects, the transient intermediate is identical with the equilibrium one, demonstrating the validity of the use of the equilibrium intermediate as a model of the folding intermediate. Essentially the same transient intermediate was also detected in the folding of lysozyme, the protein known to be homologous to alpha-lactalbumin but whose equilibrium unfolding is represented as a two-state reaction. The stability and cooperativity of the secondary structure of the intermediate of lysozyme are compared with those of alpha-lactalbumin. The results show that the protein folding occurring via the intermediate is not limited to the proteins that show equilibrium intermediates. Although the unfolding equilibria of most proteins are well approximated as a two-state reaction, the two-state hypothesis may not be applicable to the folding reaction under the native condition. Two models of protein folding, intermediate-controlled folding model and multiple-pathway folding model, which are different in view of the role of the intermediate in determining the pathway of folding, are also discussed.

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We have developed a multimodal complementary metal-oxide-semiconductor (CMOS) sensor device for observing neural activities in the deep brain of a mouse. The CMOS sensor includes an image sensor, electrodes, and a light-emitting diode (LED). The image sensor was designed to be operated using only four inputs/outputs (I/Os) to reduce the number of connecting wires. The electrodes were placed on the pixel array of the sensor. Windows were opened in the electrode over the photodiodes to enable the fluorescence to be imaged using the pixels under the electrodes. An LED was mounted on the chip. The sensor chip was shaped like a shank to facilitate smooth insertion into the brain tissue. The entire device was coated with a parylene layer to make it biocompatible. The experimental results showed that the green fluorescent beads on the pixel array were successfully imaged using the LED on the chip as a light source. In a brain phantom, the change in the electrical potential was successfully sensed by the electrode, and green fluorescent beads were simultaneously imaged using the pixels under the electrode. We also demonstrated that the CMOS sensor device could successfully operate in the hippocampal area of an anesthetized mouse.

Thin-film transistors (TFTs) were fabricated on polycrystalline silicon (poly-Si) films formed by position-controlled largegrain growth technology using an excimer laser. The field-effect mobility, on-off transition slope, and threshold voltage were 914 cm 2 V À1 s À1 , 93 mV/decade, and 0.58 V for the n-channel device, and 254 cm 2 V À1 s À1 , 122 mV/decade, and À0:43 V for the p-channel device, respectively. These values indicate that TFTs had an ultrahigh performance comparable to that of {100}-oriented crystal-silicon metal-oxide-semiconductor (MOS) transistors. Furthermore, their effective mobilities had the same effective field and temperature dependences as those of MOS transistors, indicating that electrons and holes were predominantly scattered not by random grain boundaries or defects in the Si film, but by phonons at the SiO 2 -Si interface, similarly to those of crystal-silicon MOS transistors. These attractive results were obtained as a result of the fact that the TFT channel region was made up of nearly {100}-oriented single grains.