Development and validation of the AmpFlSTR Yfiler PCR amplification kit: a male specific, single amplification 17 Y-STR multiplex system

In the past 5 years, there has been a substantial increase in the use of Y-short tandem repeat loci (Y-STRs) in forensic laboratories, especially in cases where typing autosomal STRs has met with limited success. The AmpFlSTR Yfiler PCR amplification kit simultaneously amplifies 17 Y-STR loci including the loci in the "European minimal haplotype" (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, and DYS393), the Scientific Working Group on DNA Analysis Methods (SWGDAM) recommended Y-STR loci (DYS438 and DYS439), and the highly polymorphic loci DYS437, DYS448, DYS456, DYS458, Y GATA H4, and DYS635 (formerly known as Y GATA C4). The Yfiler kit was validated according to the FBI/National Standards and SWGDAM guidelines. Our results showed that full profiles are attainable with low levels of male DNA (below 125 pg) and that under optimized conditions, no detectable cross-reactive products were obtained on human female DNA, bacteria, and commonly encountered animal species. Additionally, we demonstrated the ability to detect male specific profiles in admixed male and female blood samples at a ratio of 1:1000.     

Identification of a novel polymorphism in the X-chromosome region homologous to the DYS456 locus

During an extensive multipopulation study with Y-short tandem repeat (STR) loci, amplified using the AmpFlSTR Yfiler PCR amplification kit, amplification of a 71 bp fragment was observed in 2.32% of the male samples analyzed (N = 3141). By direct sequencing of this fragment, it was determined that the primer binding sequences were identical to those of the DYS456 locus. A T to G single-nucleotide polymorphism (SNP) enabled amplification of the 71 bp fragment. The SNP is located within an X-Y homologous region at Xq21.31 and was observed with the highest frequency within the African American and Sub-Saharan African populations in our study. Presence of SNP on the X chromosome did not interfere with the reliability of typing the DYS456 locus and the other Y-STR loci typeable using the AmpFlSTR Yfiler PCR amplification kit. Full profiles in a mixture of male:female at 1:4000 were obtained using the current configuration of the AmpFlSTR Yfiler kit even in the presence of female DNA containing the G variant.     

Developmental validation of the AmpF?STR SEfiler Plus™ PCR amplification kit: An improved multiplex with enhanced performance for inhibited samples

Since its introduction in 2002, the AmpF?STR^® SEfiler™ kit has provided a highly discriminating DNA profiling option to German forensic laboratories by combining the widely used SGM Plus^® Kit loci with the SE-33 locus required for the German DNA Database. Whilst proving successful on database samples, laboratories using the SEfiler™ kit have reported the need for chemistry better able to handle the ever-increasing number of casework samples.The new AmpF?STR^® SEfiler Plus™ kit contains the same loci and primer sequences as the SEfiler™ kit but uses improved synthesis and purification processes to minimize the presence of dye-labeled artifacts. Other improvements include modified PCR cycling conditions for enhanced sensitivity and a new buffer formulation that improves performance with inhibited samples when compared to the original SEfiler™ kit.Validation studies demonstrating the effectiveness of the multiplex are presented with emphasis on the models of inhibition and casework samples.     

The Association between Maternal Exposure to Intimate Partner Violence and Emotional and Behavioral Problems in Spanish Children and Adolescents

This correlational cross-sectional study was designed to investigate whether the intimate partner violence (IPV) suffered by mothers (physical and psychological maltreatment), the neglect suffered by children, and the maltreatment (physical and psychological) directly suffered by children are statistically associated with an increase in the probability of the child’s suffering psychopathological problems. The sample consisted of 189 Spanish children aged 6 to 17 and their mothers, recruited from Centers of Specialized Assistance for Women Victims of IPV. The results of a canonical correlation analysis showed that the most significant problems suffered by the children were both externalizing and internalizing ones. In girls, the maltreatment suffered by their mothers was directly related to a larger frequency of somatic complaints than in boys. In addition, physical maltreatment to the mother and emotional maltreatment suffered by the child exhibited a statistically significant relationship with aggressive behaviour, thought problems, rule-breaking behaviour, attention problems, and withdrawn-depressed.     

Should I report? The role of general and sexual orientation-specific bullying policies and teacher behavior on adolescents’ reporting of victimization experiences

Empowering students to report peer victimization is a major part of bullying intervention. Nevertheless, although bullying often can be discriminatory (e.g., homophobic), there remains limited attention to how issues of diversity relate to students reporting their victimization experiences. Among 572 students in four Chilean high schools, we tested a model in which multiple factors predicted students’ consistency of reporting victimization. Students who perceived having a sexual orientation/gender identity-specific antibullying policy reported less frequently hearing teachers or staff make homophobic comments, and students who perceived having a general antibullying policy reported that teachers and staff more frequently intervened against homophobic behavior. In turn, hearing teachers and staff make fewer homophobic comments and seeing them more frequently correct homophobic behavior related to students reporting stronger trust in their teachers and staff. Finally, stronger trust in teachers and staff was associated with more consistently reporting victimization experiences.     

Concordance study between the AmpFlSTR MiniFiler PCR amplification kit and conventional STR typing kits

The AmpFlSTR MiniFiler polymerase chain reaction amplification kit developed by Applied Biosystems enables size reduction on eight of the larger STR loci amplified in the Identifiler kit, which will aid recovery of information from highly degraded DNA samples. The MiniFiler Kit amplifies CSF1PO, FGA, D2S1338, D7S820, D13S317, D16S539, D18S51, and D21S11 as well as the sex-typing locus amelogenin. A total of 1308 samples were evaluated with both the MiniFiler and Identifiler STR kits: 449 African American, 445 Caucasian, 207 Hispanic, and 207 Asian individuals. Full concordance between Identifiler and MiniFiler Kits was observed in 99.7% (10,437 out of 10,464) STR allele calls compared. The 27 differences seen are listed in Table 1 and encompass the loci D13S317 (n = 14) and D16S539 (n = 10) as well as D18S51 (n = 1), D7S820 (n = 1), and CSF1PO (n = 1). Genotyping discrepancies between the Identifiler and MiniFiler kits were confirmed by reamplification of the samples and further testing using the PowerPlex 16 kit in many cases. DNA sequence analysis was also performed in order to understand the nature of the genetic variations causing the allele dropout or apparent repeat unit shift.     

The Legacy of the Maastricht-Urteil and the Pluralist Movement

Abstract:  The Maastricht-Urteil of the German Constitutional Court of October 1993 has left a deep mark on EU law. Although some may consider it as part of legal history, the decision has never been overruled, and the ideas behind it are very much alive. This article tries to examine the legacy of that decision. From a practical point of view, the article focuses on the following issues: the current situation in Germany; the influence on other constitutional or supreme courts and on constitutional reforms in some Member States; the influence on the European Court of Justice and on the Treaty establishing a Constitution for Europe. Regarding theory, three sections of the article discuss a number of widespread ' idées reçues ' contained in the Maastricht-Urteil on notions such as the state, constituent power ( pouvoir constituant ), and democracy. The next section presents the movement of legal pluralism as an attempt to come to terms with the Maastricht-Urteil and its legacy. It criticises the radical versions of legal pluralism in view of the damage they may cause to essential dimensions of the rule of law. The final section reflects on the real motives behind the Maastricht-Urteil and its legacy, and on possible future developments.