Automation of the Differential Digestion Process of Sexual Assault Evidence.

Sexual assault evidence samples require the use of a specific process known as a differential digestion to separate sperm from nonsperm cells prior to DNA extraction. An automated differential digestion process was developed using a selective degradation technique, which uses DNase I to digest the remaining nonsperm DNA in the sperm fraction. The use of DNase on pristine samples, as well as aged and degraded samples, was assessed to ensure that the quantity and quality of the sperm DNA were not compromised or adversely affected. Samples processed using the selective degradation technique yielded comparable DNA yield and DNA typing data to the conventional differential digestion process. The automated process utilized 96-well plates for high throughput and incorporated microscope slide preparations for confirmation of sperm. It reduced processing time by about sixfold and was paramount in the elimination of the Oakland Police Department Criminalistics Laboratory's sexual assault kit backlog.

High-resolution melt analysis of the minisatellite D1S80: a potential forensic screening tool.

High-resolution melt (HRM) analysis of the VNTR region of the human D1S80 locus, a 16-bp repeat minisatellite from approximately 400 to over 700 bp in length, was investigated. A Qiagen Rotor-Gene Q using the Type-it PCR HRM kit was used to acquire HRM curves for 14 single, and 16 biallelic, dsDNA samples. The HRM analysis was applicable over a range of DNA concentrations; however the characteristics of the melt curve did depend on the forward and reverse primer ratio. Despite the large amplicon size and the similarities of the repeat sequences, it was possible to discriminate different genotypes. Heterozygotes were clearly different from the homozygous variants and even small differences in the repeat sequence could be differentiated. However, the melt analysis requires a high-resolution system with temperature resolution of 0.02°C or better in order to sort out differences in these large amplicons of near identical GC content (in this case 56%). HRM analysis of amplicons with large repeat sequences can be used as a means of comparing DNA fragments. Examination of multiple sequences can be used to differentiate DNA samples and demonstrate the potential of HRM analysis as a rapid and inexpensive prescreening technique in forensic applications.

Management of mandibular rotation after a mandibulectomy: a clinical report.

Oral rehabilitation after a mandibulectomy is a challenge for prosthodontists. This clinical report describes the management of a 54-year-old man, who, after a mandibulectomy, had rotation of the mandible, with a revascularized flap, followed by an implant-supported prosthesis and ceramic restorations.

The accuracy of cephalometric tracing superimposition.

PURPOSE: The purpose of this study was to compare the accuracy of 4 methods for cephalometric tracing superimposition. They are the FH@Porion method, S-N@Sella method, least-squared averaged 5 landmarks (LS-5) method, and manual geometric method. MATERIALS AND METHODS: Eight lateral cephalometric radiographs were used. Cephalometric tracing was performed by 2 examiners. One had extensive experience in landmark digitization while the other had minimal experience. The radiographs were scanned and the reference landmarks ANS, Point A, Point B, and Pogonion were digitized, creating 8 master tracings. Then 6 digital copies of each master tracing were made, 3 for each examiner. Subsequently, the examiners were asked to digitize and trace predetermined cranial base landmarks and structures. Tracings occurred at 1-month intervals. As a result, 3 separate tracings of each set were obtained from each examiner. The tracings of each set were superimposed using 4 different methods in the CASSOS software (SoftEnable Technology Ltd, Hong Kong SAR, China). For each method of superimposition, the coordinates of ANS, Point A, Point B, and Pogonion were recorded. Their means and variances were calculated. The variance represents the variability of the superimposition method. A general linear model for repeated measures was computed to test whether there were statistically significant differences among the 4 superimposition methods, 2 examiners, 4 reference landmarks, and 2 directions. Because the distribution of the variances was skewed, they were transformed to log variances. Finally, the errors of the superimposition in millimeters for each given examiner, superimposition method, reference landmark, and direction (X, Y) were calculated. RESULTS: There was a statistically significant difference in measurement variability among the 4 superimposition methods (P < .001). For both examiners, the variability of the different superimposition methods from the highest to the lowest was: Frankfort Plane registered at Porion method, Sella-Nasion registered at Sella method, least-square averaged 5 landmarks method, and the manual geometric method. In addition, there was a statistically significant difference in the magnitude of superimposition errors between the 2 examiners (P < .001). The experienced examiner was consistently more precise than the inexperienced examiner across all methods. Moreover, there was a statistically significant difference among 4 reference landmarks (P < .001). For both examiners, the recorded variability of each given reference landmark from the lowest to the highest was: ANS, Point A, Point B, and Pogonion. Furthermore, the variability differences between horizontal and vertical directions did not reach a conventional level of significance (P = .123). Finally, the recorded errors in millimeters for each superimposition method were summarized. A smaller error in millimeters represented a higher accuracy in superimposition. The error of using manual geometric or LS-5 methods for both examiners was less than 0.50 mm, while the error of using the other 2 methods was up to 0.99 mm for the experienced examiner and 2.88 mm for the inexperienced examiner. CONCLUSION: The error of both manual and LS-5 methods was within 0.5 mm. The LS-5 method had its advantage because it could be automated by the computer.