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A simple method for extracting protein from human anagen (i.e., actively growing hair stage) head hairs was developed in this study for cases of limited sample availability and/or studies of specific micro-features within a hair. The distinct feature segments of the hair from one donor were divided lengthwise (i.e., each of ∼200–400 μm) and then pooled for three individual hairs to form a total of eight composite hair samples (i.e., each of ∼1 mm or less in total length). The proteins were extracted, digested using trypsin, and characterized via nano-flow liquid chromatography tandem-mass spectrometry (nLC-MS/MS). A total of 63 proteins were identified from all eight protein samples analyzed of which 60% were keratin and keratin-associated proteins. The major hair keratins identified are consistent with previous studies using fluorescence in situ hybridization and nLC-MS/MS while requiring over 400–8000-fold less sample. The protein extraction method from micro-sized human head hairs described in this study will enable proteomic analysis of biological evidence for cases of limited sample availability and will complement hair research. For example, research seeking to develop alternative non-DNA based techniques for comparing questioned to known hairs, and understanding the biochemistry of hair decomposition.

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Objectives: Analysis of a mass burial of seven males at CA-ALA-554, a prehistoric site in the Amador Valley, CA, was undertaken to determine if the individuals were "locals" or "non-locals," and how they were genetically related to one another.Methods: The study includes osteological, genetic (mtDNA), and stable (C, N, O, S) and radiogenic (Sr) isotope analyses of bone and tooth (first and third molars) samples.Results: Isotopes in first molars, third molars, and bone show they spent the majority of their lives living together. They are not locals to the Amador Valley, but were recently living to the east in the San Joaquin Valley, suggesting intergroup warfare as the cause of death. The men were not maternally related, but represent at least four different matrilines. The men also changed residence as a group between age 16 and adult years.Conclusions: Isotope data suggest intergroup warfare accounts for the mass burial. Genetic data suggest the raiding party included sets of unrelated men, perhaps from different households. Generalizing from this case and others like it, we hypothesize that competition over territory was a major factor behind ancient warfare in Central California. We present a testable model of demographic expansion, wherein villages in high-population-density areas frequently fissioned, with groups of individuals moving to lower-population-density areas to establish new villages. This model is consistent with previous models of linguistic expansion. Am J Phys Anthropol 159:116-125, 2016. V C 2015 Wiley Periodicals, Inc.A romanticized interpretation often depicts huntergatherers as peaceful peoples with, at most, very low rates of interpersonal violence when compared with more complex and state-level societies. Recent archaeological and anthropological research has challenged these notions (e.g., Walker, 1989;Keeley, 1996;LeBlanc, 1999;Lambert, 2002;Arkush and Allen, 2006; Allen, 2012;Jones and Allen, 2014). These studies suggest that the level of organization in warfare was often lower among hunter-gatherers, but that per-capita rates of non-lethal violence and homicide were often just as high (but see Fry, 2007 andFry andS€ oderberg, 2013 for an alternate perspective).The ancient hunter-gatherers of California have played an important part in this shift in thinking (Walker, 1989;Jurmain, 1991Jurmain, , 2001Jurmain and Bellifemine, 1997;Lambert, 1997Lambert, , 2007Andrushko et al., 2005Andrushko et al., , 2010Jurmain et al., 2009;Bartelink et al., 2013; Eerkens et al., 2014a,b;Schwitalla et al., 2014). Osteological studies show that violence often played a role in mortality. A recent compilation and survey of osteological data (Schwitalla et al., 2014) shows that sharp-force trauma appears in over 7% and blunt-force in over 4% of individuals

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Neither microscopical hair comparisons nor mitochondrial DNA sequencing alone, or together, constitutes a basis for personal identification. Due to these limitations, a complementary technique to compare questioned and known hair shafts was investigated. Recently, scientists from Lawrence Livermore National Laboratory's Forensic Science Center and other collaborators developed a peptide profiling technique, which can infer nonsynonymous single nucleotide polymorphisms (SNPs) preserved in hair shaft proteins as single amino acid polymorphisms (SAPs). In this study, peptide profiling was evaluated to determine if it can meet forensic expectations when samples are in limited quantities with the possibility that hair samples collected from different areas of a single donor's scalp (i.e., single source) might not exhibit the same SAP profile. The average dissimilarity, percent differences in SAP profiles within each source, ranged from 0% difference to 29%. This pilot study suggests that more work is needed before peptide profiling of hair can be considered for forensic comparisons.

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