Genetic sexing to determine the optimal discriminant functions for the analysis of archaeological remains from El Hierro (Canary Islands)

A correct sex assignment of a given bone or bone fragment is of paramount importance for the archaeologist, anthropologist and in forensic medicine. Discriminant functions, combining several anthropometric measurements obtained from individuals with known sex are useful tools for this purpose, but it is essential to know exactly the sex from which the measures are obtained. This is an easy task in modern populations, but it is problematic in ancient ones, since even when the entire skeleton is available, diagnosis of sex is not 100% accurate. Sexing by genetic methods by amplifying the first intron of the amelogenin gene constitutes a much more accurate method for sexing bones and may be the gold standard for further elaboration of discriminant functions which may serve for sexing new bones dug up in future excavations. With this aim we have genetically sexed 52 (out of 59) tibiae belonging to the prehispanic population of El Hierro, in the Canary Islands, identifying 18 women and 34 men, and then, performed discriminant functions combining several anthropometric variables. These functions show a high accuracy in sex diagnosis (94.2%; area under ROC curve = 0.954 with the best of the functions), so that they allow correct sexing of tibiae or tibiae fragments (only proximal third, distal third or midshaft). Thus, genetic sexing obviates the problem of finding an accurate gold standard for the elaboration of discriminant functions for ancient bones. This method could be applied to other populations of different antiquity and different ethnicity.     

Canary islands aborigin sex determination based on mandible parameters contrasted by amelogenin analysis

Sex determination using mandible parameters is population dependent. In order to assess which measurements better characterize sex in prehispanic individuals from the Canary Islands, we blindly contrasted the results obtained by visual inspection and osteometric measurements with those obtained by molecular sexing using amelogenin ancient DNA analysis on teeth from the same material. Unambiguous sex classification was achieved by amplification of sex specific amelogenin alleles in 56 out of 76 mandibles (73.78% of the cases). Visual inspection led to a correct diagnosis in 66.04% of cases, with a greater proportion of errors for female (54.17%) than male (17.24%) mandibles. Osteometric measurements were able to assign sex correctly in 72.2% in the best of cases (mandibular height), a proportion similar to that obtained using a discriminant function (71.2%). By logistic regression analysis, ramus breadth, index ramus breadth/ramus height and mandibular length were the parameters independently related with a mistaken diagnosis of female sex, whereas bigonial width, ramus height and mandibular length were the parameters more closely and independently related to a mistaken diagnosis of male sex. In conclusion, diagnosis based on visual examination of the mandible or on its metric measurement only serves to roughly estimate sex with an accuracy of around 70% or less, at least among the prehispanic population from Gran Canaria. Amplification of amelogenin alleles leads to unambiguous identification of male and female alleles in 73.68% of cases, at least among the prehispanic population from Gran Canaria.     

Sex identification of ancient DNA samples using a microfluidic device

A microfluidic device has been developed for the sex identification of ancient DNA samples and works by manipulating liquids within an environment of micrometer dimensions. In this work a range of microfluidic DNA extraction methods were evaluated for their compatibility with ancient DNA samples, and the use of streptavidin-coated super paramagnetic particles to isolate biotin-labeled abasic sites within damaged DNA was shown to be the most reproducible. Polymerase chain reaction-based DNA amplification was possible on the microfluidic device when less than 50 pg of template DNA was added. As a proof-of-principle, powdered bone samples were analysed using the integrated methodology developed. Following conventional capillary gel electrophoresis, two out of the three samples produced positive amplification results and were successfully identified as female. These sex identifications were corroborated by independent Amelogenin, anthropological and Y chromosome analysis. The work reported here is the first step in the development of a complete miniaturized microfluidic system that would enable on-site ancient DNA analysis.     

Dietary patterns during the early prehispanic settlement in La Gomera (Canary Islands)

The dietary pattern of 10 adults interred in the Acceso al Pescante de Vallehermoso cave (La Gomera, Canary Islands), dated from 1600 to 1800 years BP, has been investigated using carbon and nitrogen stable isotope signatures, bone barium and strontium levels, and dental and oral pathologies. In addition, trabecular bone mass – as a parameter useful to evaluate overall nutrition – was also determined.