Helicases are motor proteins that are involved in DNA and RNA metabolism, replication, recombination, transcription and repair. The motors are powered by ATP binding and hydrolysis. Hepatitis C virus encodes a helicase called non-structural protein (NS3). NS3 possesses protease and helicase activities on its N-terminal and C-terminal domains respectively. The helicase domain of NS3 protein is referred as NS3h. In vitro, NS3h catalyzes RNA and DNA unwinding in a 3' to -5' direction. The directionality for unwinding is thought to arise in part from the enzyme's ability to translocate along DNA, but translocation has not been shown explicitly. We examined the DNA translocase activity of NS3h by using single-stranded oligonucleotide substrates containing a fluorescent probe on the 5' end. NS3h can bind to the ssDNA and in the presence of ATP, move towards the 5'-end. When the enzyme encounters the fluorescent probe, a fluorescence change is observed that allows translocation to be characterized. Under conditions that favor binding of one NS3h per DNA substrate (100 nM NS3h, 200 nM oligonucleotide) we find that NS3h translocates on ssDNA at a rate of 46 ± 5 nt s −1 and that it can move for 230 ± 60 nt before dissociating from the DNA. The translocase activity of some helicases is responsible for displacing proteins that are bound to DNA. We studied protein displacement by using a ssDNA oligonucleotide covalently linked to biotin on the 5'-end. Upon addition of streptavidin, a 'protein-block' was placed in the pathway of the helicase. Interestingly, NS3h was unable to displace streptavidin from the end of the oligonucleotide, despite its ability to translocate along the DNA. The DNA unwinding activity of NS3h was examined using a 22 bp duplex DNA substrate under conditions that were identical to † This work was supported by NIH research grants R24 GM080599 (K.D.R.) and R01 GM089001 (K.D.R and C.E.C.). *Corresponding authors: Kevin D. Raney, Tel: (501) raneykevind@uams.edu; Christopher J. Fischer,; shark@ku.edu.. $ These authors contributed equally to this work. SUPPORTING INFORMATION AVAILABLESupplementary file provides data on size exclusion chromatography of NS3h in the presence of ssDNA, NS3 translocation on 5'-F-(dT) L, NS3 translocation on Poly(dT)/M13, NS3h translocation on Cy3-labeled oligos, NS3h translocation in the presence of different concentrations of heparin, and NS3h translocation on biotinylated oligo (3'T58BiodTG5') in the presence and absence of streptavidin. The supporting information is available for free of charge via the internet at

Forensic casework from past-conflicts relies on the corrected historical Trotter data for stature estimation in Fordisc. For roughly 10 years', stature estimation using this data has produced point estimates for the tibia that are on average 1.25 inches less than the other long bones. This issue was identified after applying the equations derived from Fordisc to the USS Oklahoma commingled assemblage. Reevaluation of Fordisc revealed that a correction factor of 20 mm, instead of 10 mm, was mistakenly applied to the Trotter tibia data. Historical forensic anthropology reports written at the Defense POW/MIA Accounting Agency were utilized to identify that the overcorrection is isolated to Fordisc 3 with an error rate of 5% of known antemortem statures falling outside of the prediction intervals that relied on the tibia. Further evaluation of the Oklahoma sample indicates the 10 mm correction is still producing point estimates less than the other long bones.

Large commingled assemblages present unique challenges for individual identification, because the segregation of remains into discrete individuals can be difficult and timeconsuming. These assemblages can be problematic from data management and analytical perspectives, and often an increase in the number of casualties leads to a decrease in the discriminatory power of anthropological techniques such as visual pair matching and articulation (SWGANTH 2013). These challenges can lead to lengthy analytical timeframes and delays in identification, and a multidisciplinary approach to resolving commingling is both espoused and practiced (e.g.

Estimation of the number of individuals in an assemblage is critical to determine the scale of an incident and whether all