Validation of a male-specific, 12-locus fluorescent short tandem repeat (STR) multiplex

Y chromosome-specific short tandem repeat (Y-STR) analysis has become another widely accepted tool for human identification. The PowerPlex Y System is a fluorescent multiplex that includes the 12 loci: DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439. This panel of markers incorporates the 9-locus European minimal haplotype (EMH) loci recommended by the International Y-STR User Group and the 11-locus set recommended by the Scientific Working Group on DNA Analysis Methods (SWGDAM). Described here are inter-laboratory results from 17 developmental validation studies of the PowerPlex Y System and include the following results: (a) samples distributed between laboratories and commercial standards produced expected and reproducible haplotypes; (b) use of common amplification and detection instruments were successfully demonstrated; (c) full profiles were obtained with standard 30 and 32 cycle amplification protocols and cycle number (24-28 cycles) could be modified to match different substrates (such as direct amplification of FTA paper); (d) complete profiles were observed with reaction volumes from 6.25 to 50 microL; (e) minimal impact was observed with variation of enzyme concentration; (f) full haplotypes were observed with 0.5-2x primer concentrations; however, relative yield between loci varied with concentration; (g) reduction of magnesium to 1mM (1.5 mM standard) resulted in minimal amplification, while only partial loss of yield was observed with 1.25 mM magnesium; (h) decreasing the annealing temperature by 2-4 degrees C did not generate artifacts or locus dropout and most laboratories observed full amplification with the annealing temperature increased by 2 degrees C and significant locus dropout with a 4 degrees C increase in annealing temperature; (i) amplification of individual loci with primers used in the multiplex produced the same alleles as observed with the multiplex amplification; (j) all laboratories observed full amplification with >or = 125 pg of male template with partial and/or complete profiles observed using 30-62.5 pg of DNA; (k) analysis of < or = 500 ng of female DNA did not yield amplification products; (l) the minor male component of a male/female mixture was observed with < or =1200-fold excess female DNA with the majority of alleles still observed with 10,000-fold excess female; (m) male/male mixtures produced full profiles from the minor contributor with 10-20-fold excess of the major contributor; (n) average stutter for each locus; (o) precision of sizing were determined; (p) human-specificity studies displayed amplification products only with some primate samples; and (q) reanalysis of 102 non-probative casework samples from 65 cases produced results consistent with original findings and in some instances additional identification of a minor male contributor to a male/female mixture was obtained. In general, the PowerPlex Y System was shown to have the sensitivity, specificity and reliability required for forensic DNA analysis.     

Validation of male-specific, 12-locus fluorescent short tandem repeat (STR) multiplex

Y chromosome-specific short tandem repeat (Y-STR) analysis has become another widely accepted tool for human identification. The PowerPlex Y System is a fluorescent multiplex that includes the 12 loci: DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439. This panel of markers incorporates the 9-locus European minimal haplotype (EMH) loci recommended by the International Y-STR User Group and the 11-locus set recommended by the Scientific Working Group on DNA Analysis Methods (SWGDAM). Described here are inter-laboratory results from 17 developmental validation studies of the PowerPlex Y System and include the following results: (a) samples distributed between laboratories and commercial standards produced expected and reproducible haplotypes; (b) use of common amplification and detection instruments were successfully demonstrated; (c) full profiles were obtained with standard 30 and 32 cycle amplification protocols and cycle number (24-28 cycles) could be modified to match different substrates (such as direct amplification of FTA paper); (d) complete profiles were observed with reaction volumes from 6.25 to 50 microL; (e) minimal impact was observed with variation of enzyme concentration; (f) full haplotypes were observed with 0.5-2x primer concentrations; however, relative yield between loci varied with concentration; (g) reduction of magnesium to 1mM (1.5 mM standard) resulted in minimal amplification, while only partial loss of yield was observed with 1.25 mM magnesium; (h) decreasing the annealing temperature by 2-4 degrees C did not generate artifacts or locus dropout and most laboratories observed full amplification with the annealing temperature increased by 2 degrees C and significant locus dropout with a 4 degrees C increase in annealing temperature; (i) amplification of individual loci with primers used in the multiplex produced the same alleles as observed with the multiplex amplification; (j) all laboratories observed full amplification with >or = 125 pg of male template with partial and/or complete profiles observed using 30-62.5 pg of DNA; (k) analysis of < or = 500 ng of female DNA did not yield amplification products; (l) the minor male component of a male/female mixture was observed with < or =1200-fold excess female DNA with the majority of alleles still observed with 10,000-fold excess female; (m) male/male mixtures produced full profiles from the minor contributor with 10-20-fold excess of the major contributor; (n) average stutter for each locus; (o) precision of sizing were determined; (p) human-specificity studies displayed amplification products only with some primate samples; and (q) reanalysis of 102 non-probative casework samples from 65 cases produced results consistent with original findings and in some instances additional identification of a minor male contributor to a male/female mixture was obtained. In general, the PowerPlex Y System was shown to have the sensitivity, specificity and reliability required for forensic DNA analysis.     

Media Framing on Armed Conflicts: Limits of Peace Journalism on the Nagorno-Karabakh Conflict

ABSTRACT

Media framing and coverage of contemporary armed conflicts largely focus on the defence of national security and war propaganda. The concept of peace journalism draws on the Galtung tradition and provides a toolkit for opposing war journalism and contributing to reconciliation. By examining the case of media framing of the Nagorno-Karabakh conflict, this article asks whether and why peace journalism is possible in the given political context. Our study of limited peace journalism shows that traditions of war journalism are predominant and points to limitations of liberal instruments in (semi-)authoritarian post-Soviet conflict contexts.     

Community Involvement in Rural Regeneration Partnerships: Exploring the Rural Dimension

This paper is about the involvement of local communities in rural regeneration and area development partnerships. In particular it asks questions about the extent to which there are distinctive rural elements to this which differentiate and counterpoise it to such involvement in urban areas. The paper concludes that, whilst there are some commonalities between such involvement across urban and rural communities, there are also challenges particular to rural areas. These are the spatial features of rural areas, the paucity of human capital in rural communities and a much lower resource level than in urban communities. The paper concludes that these challenges can only be met effectively in rural areas where there is strong local voluntary and community sector infrastructure to support community involvement in rural regeneration partnerships.     

Military professionalism in post‐communist Hungary and Poland: An analysis and assessment

Abstract

This article discusses Russian perceptions of and attitudes toward the North Atlantic Treaty Organization (NATO). Russia has historically disliked and mistrusted NATO, seeing it as the primary threat to its international aspirations; in practice Russia pursues a dual policy. Its harsh condemnation of NATO has not stopped it from cooperating in selected areas of mutual interest. The most important among them is support for NATO's military operations in Afghanistan. The recent rejuvenation of relations between the west and Moscow is known as the strategic ‘reset’, meaning a return to diplomatic contacts and limited cooperation regardless of disagreements over the invasion of Georgia and Moscow's other recent international transgressions. The reset in NATO–Russia relations has only tactical significance, however. Cooperation will take place on a limited basis, but a genuine reset in mutual relations must wait for a reset in Russia's political and strategic priorities.     

Path-Dependent Institutions and Strategic Veto Players: National Parliaments in the European Union

According to the prevailing concept of 'dual legitimacy', national parliaments constitute an important source of democratic legitimacy in the EU. Reinforced parliamentary scrutiny and control of the national representatives in the Council of Ministers seem to contribute to a more democratic Europe. However, if parliaments tie the hands of their governments when they negotiate at the European level, effectiveness of policy-making is jeopardised and national interests may be defeated. Realising this dilemma, members of national parliaments develop strategies to deal with conflicting requirements of national party politics and European policy-making. These strategies and their implications for democracy are influenced by the path-dependent institutional changes in national parliamentary systems. They therefore vary considerably between member states.