Optimal processing for proteomic genotyping of single human hairs.

The use of hair evidence for human identification is undergoing considerable improvement through the adoption of proteomic genotyping. Unlike traditional microscopic comparisons, protein sequencing provides quantitative and empirically based estimates for random match probability. Non-synonymous SNPs are translated as single amino acid polymorphisms and result in genetically variant peptides. Using high resolution mass spectrometry, these peptides can be detected in hair shaft proteins and used to infer the genotypes of corresponding SNP alleles. We describe experiments to optimize the proteomic genotyping approach to individual identification from a single human scalp hair 2 cm in length (∼100 µg). This is a necessary step to develop a protocol that will be useful to forensic investigators. To increase peptide yield from hair, and to maximize genetically variant peptide and ancestral information, we examined the conditions for reduction, alkylation, and protein digestion that specifically address the distinctive chemistry of the hair shaft. Results indicate that optimal conditions for proteomic analysis of a single human hair include 6 h of reduction with 100 mM dithiothreitol at room temperature, alkylation with 200 mM iodoacetamide for 45 min, and 6 h of digestion with two 1:50 (enzyme:protein) additions of stabilized trypsin at room temperature, with stirring incorporated into all three steps. Our final conditions using optimized temperatures and incubation times increased the average number of genetically variant peptides from 20 ±â€¯5 to 73 ±â€¯5 (p = 1 × 10-13), excluding intractable hair samples. Random match probabilities reached up to 1 in 620 million from a single hair with a median value of 1 in 1.1 million, compared to a maximum random match probability of 1 in 1380 and a median value of 1 in 24 for the original hair protein extraction method. Ancestral information was also present in the data. While the number of genetically variant peptides detected were equivalent for both European and African subjects, the estimated random match probabilities for inferred genotypes of European subjects were considerably smaller in African reference populations and vice versa, resulting in a difference in likelihood ratios of 6.8 orders of magnitude. This research will assure uniformity in results across different biogeographic backgrounds and enhance the use of novel peptide analysis in forensic science by helping to optimize genetically variant peptide yields and discovery. This work also introduces two algorithms, GVP Finder and GVP Scout, which facilitate searches, calculate random match probabilities, and aid in discovery of genetically variant peptides.

Influence of different cosmetic vehicles in mechanical and physical properties of hair treated with oxidative hair dyes

ABSTRACT Hair care products play a significant role in the cosmetic market and aim at improving hair brightness, breakage resistance, and color change. In this study, we analyzed the possibility of the formulation of oxidative dyes in different vehicles impacting the hair's both mechanical and physical properties. Light brown and light blond dyes were prepared using eight different vehicles. Among these, four vehicles were emulsifying agents and four gelling agents. Each formulation was applied to two types of virgin Caucasian hair (light blond and dark brown). Physical, chemical, and organoleptic properties of each formulation were assessed, as well as changes in hair parameters after oxidative dyeing, such as staining intensity, brightness, and breaking strength. The parameters of color and brightness differed in some formulations, but the hair type also responded differently. Brightness parameter was increased in dark brown hair colored with both dyes, whereas light blond hair showed the opposite result. Regarding the breaking strength, there were no significant differences between the two kinds of tresses. Cosmetic formulations should adjust the consumer desired effects (e.g. color change) in order to present minimal drawbacks (e.g. decrease of hair brightness and strength). Thus, the study of different vehicles is important when establishing the best oxidative dye formulation.