Automation and Integration in Semiconductor Manufacturing

Targets 32 2. Materials and methods Surgical procedureMale Wistar rats (89 weeks old, 200250g in weight) were anesthetized by intraperitoneal pentobarbital (50mg/kg) injection.The scalp was incised on the midline and the skull was exposed.A 22.5mm hole was drilled (1 mm posterior, + 1 mm right lateral to bregma) in the right parietal calvaria.Brain injury above the dura mater was then induced with a pneumatic control injury device (Itoh et al., 2005; Itoh et al., 2007;Itoh et al., 2009) at an impact velocity of 4m/sec (impact tip diameter of 1mm; fixed impact deformation of 2mm depth from the cerebral surface) (Itoh et al., 2005; Itoh et al., 2007;Itoh et al., 2009).Control rats were subjected to a sham operation, but no cortical penetration injury was inflicted.The contralateral hemisphere was not used as a control, since that area may have been affected by the impact.Rats were placed in a heated cage to maintain their body temperature at 37°C during the recovery from anesthesia. Anti-APP antibody infusionThe drug infusion protocol was as previously described (Green-Sadan et al., 2003).Immediately, after TBI, the animals were placed in a stereotactic frame and fitted with a brain infusion cannula (Alzet brain infusion kit 3, 1mm, Alzet, San Diego, CA, USA).The cannula was implanted with the tip inserted into the damaged brain region (1 mm posterior, + 1 mm right lateral to bregma).The cannula was secured with dental cement.A microosmotic pump (Model 2001; rate of 1μl/h, Alzet), filled with 300 μl infusion volume kept 37°C, was implanted subcutaneously in the neck and connected to the infusion cannula.Rats received an infusion of either anti-APP antibody (IgG2a isoform, 1μg/ml; anti-APP antibody group; Chemicon, Temecula, CA, USA) diluted in sterilized phosphate buffer saline (0.1M PBS, pH 7.47.5;n=20) or sterilized PBS only (PBS group; n=20) for seven days immediately after TBI.For the control of the Morris water maze experiments, ten animals with no operation (sham operation group, no injury) were also collected.In sham operation group, a micro-osmotic pump was implanted subcutaneously in the neck without infusion cannula.Moreover, sham operation group was used only the behavioural experiments.As probe and the extent of APP antibody infusion test, although data not shown in this study, we investigated that the extent of APP antibody diffusion with Evans blue using the osmotic pump and confirmed Evans blue extended whole brain from center of infusion area. Morris water maze experimentsA circular, thermostatically regulated, dark gray PVC-plastic water tank (180cm wide, 45cm deep, filled with tap water at 22±1°C), located in the center of the testing room and surrounded by extra-maze cues, was used in the spatial learning task.A constant asymmetrical array of lamps and pictures served as cues for spatial orientation.A circular dark gray platform (15cm wide) submerged 1cm below the water surface served as a platform.The platform was placed in the center of one of the quadrants, i.e., the target quadrant, of the water maze (Elvander et al., 2004).The experiments were monitored using a digital TV system connected to a computer (Elvander et al., 2004).Training took place between 8:00a.m. and 3:00p.m. during seven consecutive days.Each daily training session consisted of four trials with a 120s cutoff time, followed by 30s rest on the platform.Memory was tested in fifteen animals from each of the anti-APP antibody group and the PBS group, and in ten sham operation group animals, starting at day one and continued up to seven days after TBI/sham operation.www.intechopen.