The endocranial surface close to the frontal crest of the frontal bone a thickened area with one large (1065 mm) and two minor (565 mm) dense, protruding, round nodules with discrete margins was observed (Figure 3). The largest nodule, located on the left side of the frontal crest, protruded approximately 4 mm. The surface of the thickened area is smooth, but the endocranial tabula exhibits clusters of swollen billows superior to the nodules, foremost close to the right side of the coronal suture. The outer table exhibits no pathological changes.Analysis of nDNAThe sex determination by pyrosequencing analysis was consistent with the anthropological data and revealed that the cranium and ulna originate from a female individual. In addition, the paraffin-embedded tissue sample from Carin Goring’s son was sex ?determined and confirmed to originate from a male individual (Figure 5). In general, analysis of mtDNA has low discrimination capacity, and the haplotype in this particular case is very common. Therefore, an nDNA analysis was attempted to further improve the evidentiary value. A first test, using the AmpliSTR Identifiler (STR) Kit from Applied Biosystems, failed to yield results from the remains. Further analysis was performed using the STR markers TPOX, TH01, D5S818, D7S820 and D8S1179, which were amplified in short targets to facilitate analysis of aged and degraded samples [20]. Several products for the TH01, D7S820 and D8S1179 markers could be obtained from both 
samples for further pyrosequencing analysis and genotyping. The TPOX and D5S81 markers did not amplify in either the remains or the reference sample and were therefore excluded in further analysis.?Identification of Carin GoringTable 2. Bone preservation and measurements.Element Frontal bone Occipital bone Parietal bone (left and right) Right clavicle Left claviclePreservation ( of complete bone) 100Measurements (mm)20 (area around Bregma) 100 100 GL 135.0 ant-post 13.13 inf-sup 8.Right scapula Left scapula Left radius100 100 100Cav. glen. 36.5 Cav. glen. 33.5 GL 244.0 ant-post 10.75 med-lat 15.Left ulna Left humerusc. 25 c. 75 Max Mid 20.01 Min Mid 19.90 GdB 57.Cervical vertebra x1 Thoracic vertebrae x3 Lumbar vertebrae x100 100 one 100 , one fragmented (40 )(GL = greatest length, ant-post = anterior-posterior diameter, inf-sup = inferior-superior diameter, med-lat = medial-lateral diameter, Max Mid = maximal diameter at midshaft, Min Mid = minimal diameter at midshaft, GdB = greatest distal breadth). doi:10.1371/journal.pone.0044366.tThe results for the amplified and sequenced markers are summarised in Table 3. The two samples share an allele for each of the three markers supporting a mother and son relationship. Statistical evaluation for the STR markers BTZ-043 resulted in a combined likelihood ratio (LR) of 26.9 in favour of the hypothesis that the remains are from the mother of the reference individual. When the mtDNA data is added, the LR is 12926553 increased to 277 (Table 3).DiscussionAn anthropological analysis was made of the bone elements for a thorough primary documentation of the skeletal preservation and of macroscopic bone changes. Homotaurine site Moreover, mitochondrial DNA analysis, a molecular sex determination, and nuclear DNA typing were performed, which all support the notion that the bones discovered in the Schorfheide Forest in 1991 are the remains of Carin Goring. ?The bone changes of the frontal bone demonstrate a close resemblance with hyperostosis frontalis interna (.The endocranial surface close to the frontal crest of the frontal bone a thickened area with one large (1065 mm) and two minor (565 mm) dense, protruding, round nodules with discrete margins was observed (Figure 3). The largest nodule, located on the left side of the frontal crest, protruded approximately 4 mm. The surface of the thickened area is smooth, but the endocranial tabula exhibits clusters of swollen billows superior to the nodules, foremost close to the right side of the coronal suture. The outer table exhibits no pathological changes.Analysis of nDNAThe sex determination by pyrosequencing analysis was consistent with the anthropological data and revealed that the cranium and ulna originate from a female individual. In addition, the paraffin-embedded tissue sample from Carin Goring’s son was sex ?determined and confirmed to originate from a male individual (Figure 5). In general, analysis of mtDNA has low discrimination capacity, and the haplotype in this particular case is very common. Therefore, an nDNA analysis was attempted to further improve the evidentiary value. A first test, using the AmpliSTR Identifiler (STR) Kit from Applied Biosystems, failed to yield results from the remains. Further analysis was performed using the STR markers TPOX, TH01, D5S818, D7S820 and D8S1179, which were amplified in short targets to facilitate analysis of aged and degraded samples [20]. Several products for the TH01, D7S820 and D8S1179 markers could be obtained from both samples for further pyrosequencing analysis and genotyping. The TPOX and D5S81 markers did not amplify in either the remains or the reference sample and were therefore excluded in further analysis.?Identification of Carin GoringTable 2. Bone preservation and measurements.Element Frontal bone Occipital bone Parietal bone (left and right) Right clavicle Left claviclePreservation ( of complete bone) 100Measurements (mm)20 (area around Bregma) 100 100 GL 135.0 ant-post 13.13 inf-sup 8.Right scapula Left scapula Left radius100 100 100Cav. glen. 36.5 Cav. glen. 33.5 GL 244.0 ant-post 10.75 med-lat 15.Left ulna Left humerusc. 25 c. 75 Max Mid 20.01 Min Mid 19.90 GdB 57.Cervical vertebra x1 Thoracic vertebrae x3 Lumbar vertebrae x100 100 one 100 , one fragmented (40 )(GL = greatest length, ant-post = anterior-posterior diameter, inf-sup = inferior-superior diameter, med-lat = medial-lateral diameter, Max Mid = maximal diameter at midshaft, Min Mid = minimal diameter at midshaft, GdB = greatest distal 
breadth). doi:10.1371/journal.pone.0044366.tThe results for the amplified and sequenced markers are summarised in Table 3. The two samples share an allele for each of the three markers supporting a mother and son relationship. Statistical evaluation for the STR markers resulted in a combined likelihood ratio (LR) of 26.9 in favour of the hypothesis that the remains are from the mother of the reference individual. When the mtDNA data is added, the LR is 12926553 increased to 277 (Table 3).DiscussionAn anthropological analysis was made of the bone elements for a thorough primary documentation of the skeletal preservation and of macroscopic bone changes. Moreover, mitochondrial DNA analysis, a molecular sex determination, and nuclear DNA typing were performed, which all support the notion that the bones discovered in the Schorfheide Forest in 1991 are the remains of Carin Goring. ?The bone changes of the frontal bone demonstrate a close resemblance with hyperostosis frontalis interna (.
