Evaluation of a method for typing the microsatellite D12S391 locus using a new primer pair and capillary electrophoresis.

We describe a modified method for typing a polymorphic microsatellite D12S391 locus by PCR using a newly designed primer pair. This primer pair produces shorter D12S391 amplified fragments (104-156 bp) than the primer pair originally described by Lareu et al. (209-261 bp). The detection system for the D12S391 locus using the new primer pair and capillary electrophoresis (CE) analysis was evaluated using various forensic samples. The typing results from 70 DNA samples using the new primer pair and the original primer pair were completely identical. One hundred twenty-five amplified fragments from D12S391 alleles were sized correctly within +/- 0.25 bp of the D12S391 allelic ladder. A rare allele, 19.3, previously found only in Caucasians, was found for the first time in a Japanese subject, and it was clearly distinguished from allele 20 by the CE analysis. This detection system was sensitive and could detect D12S391 types from 16 pg of genomic DNA, and from a minor component at a ratio of 1:10 in mixed samples. This system was more useful for the analysis of degraded DNA than was the method using the original primer pair, and could detect D12S391 types from bloodstains that had been stored for 26 years. In addition, the specificity of the method was demonstrated using nonhuman DNA.</p

Forensic study of sex determination using PCR on teeth samples.

In this study, sex determination using polymerase chain reaction (PCR) on tooth material was evaluated from the viewpoint of forensic medicine. The sensitivity of PCR for detection of the Y chromosome-specific alphoid repeat sequence and the X chromosome-specific alphoid repeat sequence was 0.5 pg of genomic DNA. Sex could be determined by PCR of DNA extracted from the pulp of 16 freshly extracted permanent teeth and dentine including the surface of the pulp cavity of 6 freshly extracted milk teeth. Sex could be determined using the pulp in all 20 teeth (10 male and 10 female) preserved at room temperature for 22 years. For the pulp of teeth stored in sea water, the sex could be determined in all 8 teeth immersed for 1 week and in 5 of 6 teeth immersed for 4 weeks. In the remaining 1 tooth, in which sex determination based on the pulp failed, the sex could be determined correctly when DNA extracted from the tooth hard tissue was examined. For teeth stored in soil, the sex could be determined accurately in all 8 teeth buried for 1 week, 7 of 8 teeth buried for 4 weeks, and in all 6 teeth buried for 8 weeks. When teeth were heated for 30 min, sex determination from the pulp was possible in all teeth heated to 100, 150, and 200 degrees C, and even in some teeth heated to 250 degrees C. When this method was applied to actual forensic cases, the sex of a mummified body estimated to have been discovered half a year to 1 year after death could be determined readily by examination of the dental pulp. In the skeletons of 2 bodies placed under water for approximately 1 year and approximately 11 years and 7 months, pulp tissues had been dissolved and lost, but sex determination was possible using DNA extracted from hard dental tissues. These results indicate that this method is useful in forensic practices for sex determination based on teeth samples.</p

Policy Controlled System and Its Model Checking

A policy is an execution rule (or constraint) for objects in a system to retain security and integrity of the system. We introduce a simple policy specification language and define its operational semantics. A new NFA construction algorithm that works in linear time is proposed and a model checking method for policy controlled system (PCS) is presented. We conducted verification of a sample PCS for hotel reservation by our automatic verification tool and the experimental results showed the efficiency of the proposed method

Safety and Efficacy of Percutaneous Pedicle Screw Placement Using a Power Tool

Introduction: This study compared traditional manual methods and power tool use with regard to the speed and accuracy of percutaneous pedicle screw (PPS) placement and determined the advantages associated with the use of power tools. Although the indication of PPS placement in minimally invasive spine stabilization (MISt) procedures has been recently expanded, there are no reports on PPS insertion using a power tool. Methods: We evaluated 35 patients who underwent PPS insertion using a power tool during MISt procedures. On one side, PPS insertion was performed using the manual (M) method, whereas on the contralateral side, insertion was performed using the power tool (P) method. We assessed the number of implanted PPSs, time taken to implant PPSs after guidewire insertion, and accuracy of PPS placement as ranked postoperatively using computed tomography images. Results: A total of 294 PPSs were inserted (147 using the M method and 147 using the P method). The mean PPS insertion time was 10.5 s using the P method and 27.4 s using the M method. The time required for inserting a screw using the P method remained consistent in the range of 10-15 s, whereas the time using the M method tended to increase from the second screw onward, with a range of 25-30 s. With regard to PPS insertion accuracy, a 2 mm or more pedicle breach was noted in 2 (1.4%) case after the P method and in 2 (1.4%) case after the M method. Conclusions: PPS placement using power tools has the potential to save the surgical time during MISt procedures