Sex assessment is one of the first steps of routine forensic anthropological examinations and it provides a crucial element to identify a set of human skeletal remains. In bioarchaeological contexts, this assessment is also important, as it helps in the reconstruction of past societies. Sex determination can be achieved by using several morphological or metric traits of the skull and postcranial skeleton, which have been found to have varying degrees of accuracy. In 1969 Phenice proposed a methodology focusing on three traits located on the pubis. These traits were described as either having a female or male morphology with ambiguity being rare. Phenice\'s method became regularly utilized as it was considered to be reliable. In 2012, Klales and colleagues published a revision of Phenice\'s method, as they found that it did not capture the variation in the expression of the three traits. Klales and co-authors created a visual ordinal scale of 1-5 for each of the three traits Phenice originally identified, thus adding three extra possible forms of expression. The purpose of the present research was to test both the original and revised methodologies on the same skeletal population in order to evaluate their suitability for the assessment of sex. The Luís Lopes Anthropological collection in Lisbon was used; 117 males and 117 females were scored using both methodologies. The results showed that the original method performed better (96.5% accuracy) than the revised method (92.7%).

Accurate sex identification is crucial for elucidating the biology of a species. In the absence of directly observable sexual characteristics, sex identification of wild fauna can be challenging, if not impossible. Molecular sexing offers a powerful alternative to morphological sexing approaches. Here, we present SeXY, a novel sex-identification pipeline, for very low-coverage shotgun sequencing data from a single individual. SeXY was designed to utilize low-effort screening data for sex identification and does not require a conspecific sex-chromosome assembly as reference. We assess the accuracy of our pipeline to data quantity by downsampling sequencing data from 100,000 to 1000 mapped reads and to reference genome selection by mapping to a variety of reference genomes of various qualities and phylogenetic distance. We show that our method is 100% accurate when mapping to a high-quality (highly contiguous N50 > 30 Mb) conspecific genome, even down to 1000 mapped reads. For lower-quality reference assemblies (N50 < 30 Mb), our method is 100% accurate with 50,000 mapped reads, regardless of reference assembly quality or phylogenetic distance. The SeXY pipeline provides several advantages over previously implemented methods; SeXY (i) requires sequencing data from only a single individual, (ii) does not require assembled conspecific sex chromosomes, or even a conspecific reference assembly, (iii) takes into account variation in coverage across the genome, and (iv) is accurate with only 1000 mapped reads in many cases.

It is common in forensic anthropology to draw inferences (e.g., inferences with respect to biological sex of human remains) using statistical models applied to anthropometric data. Commonly used models can output posterior probabilities, but a threshold is usually applied in order to obtain a classification. In the forensic-anthropology literature, there is some unease with this \"fall-off-the-cliff\" approach. Proposals have been made to exclude results that fall within a \"zone of uncertainty\", e.g., if the posterior probability for \"male\" is greater than 0.95 then the remains are classified as male, and if the posterior probability for \"male\" is less than 0.05 then the remains are classified as female, but if the posterior probability for \"male\" is between 0.05 and 0.95 the remains are not classified as either male or female. In the present paper, we propose what we believe is a simpler solution that is in line with interpretation of evidence in other branches of forensic science: implementation of the likelihood-ratio framework using relevant data, quantitative measurements, and statistical models. Statistical models that can implement this approach are already widely used in forensic anthropology. All that is required are minor modifications in the way those models are used and a change in the way practitioners and researchers think about the meaning of the output of those models. We explain how to calculate likelihood ratios using osteometric data and linear discriminant analysis, quadratic discriminant analysis, and logistic regression models. We also explain how to empirically validate likelihood-ratio models.

Lateral angle (LA) of the internal acoustic canal (IAC) at its opening at the surface of the petrous portion of the temporal bone is known in literature as a dimorphic measurement useful for sex assessment. However, its reliability is still debated. Moreover, no information is available about the possible relationship between LA and shape and size of the IAC. This study aims at assessing breadth, length and lateral angle of IAC on 100 CT scans belonging to subjects aged between 20 and 70 years, equally divided between males and females. Possible differences in the three measurements according to side and sex were assessed through two-way ANCOVA test, using three cranial measurements (distance between anterior and posterior nasal spine, upper facial height, bizygomatic breadth) as covariates (p < 0.05). Possible correlations among IAC measurements and with age were assessed through Pearson\'s test (p < 0.05). Results showed a significantly greater IAC breadth on the left side than on the right side; moreover, LA was significantly wider in females than in males and was positively correlated with IAC breadth in both males and females. In addition, LA was negatively correlated with age only in the female group. The study first showed that LA is strictly related to IAC morphology; moreover, it is significantly affected by masculinization of skeletal traits in females. Therefore, caution is suggested in using this measurement for sex assessment.

Identifying the skeletal remains of an unidentified individual is a priority for the medico-legal system because identification increases the chances of finding the person responsible and provides closure to the family. The purpose of this research was to develop a combined morphological and metric cranial sex assessment method using 3D technology that accommodates the medico-legal system, and their use of 3D models facilitates the technological transition to digitally archived skeletal collections. A total of 91 individuals of European biogeographical ancestry from the William M. Bass Donated Skeletal Collection (University of Tennessee Knoxville) were imaged using photogrammetry, turned into 3D models using Agisoft PhotoScan, and digitally evaluated using 3D Studio Max. This novel method digitally evaluated five cranial traits, including the nasal aperture height, nasal aperture width, mastoid length, the general size and architecture, and the supraorbital ridges, combining techniques that can only be done digitally with those that can be completed on the actual bone. Preliminary statistical tests demonstrate an overall accuracy rate of 90% when tested against the training sample (20 males, 20 females) and 75% when tested against the test sample (51 individuals). Although no intra- or inter-observer error rate tests were done, and further testing on other skeletal collections is necessary, this method allows forensic anthropologists to perform relatively easy point-to-point measurements, the quantification of traditionally non-quantified traits, the possibility of reproducible results, and the ability for future analyses or research.

Sex identification of ancient animal biological remains can benefit our understanding of historical population structure, demography and social behavior. Traditional methods for sex identification (e.g., osteological and morphometric comparisons) may be ineffective when animal remains are not well preserved, when sex distinguishing characteristics have not yet developed, or where organisms do not exhibit sex-associated phenotypic dimorphisms. Here we adapt a method developed for human sex determination so that it can be used to identify the sex of ancient and modern animal taxa. The method identifies sex by calculating the ratio of DNA reads aligning to the X chromosome to DNA reads aligning to autosomes (termed the Rx ratio). We tested the accuracy of this method using low coverage genomes from 15 modern elephants (Loxodonta africana) for which sex was known. We then applied this method to ancient elephant ivory samples for which sex was unknown, and describe how this method can be further adapted to the genomes of other taxa. This method may be especially useful when only low-coverage genomic data are obtainable. Furthermore, because this method relies on only the X and not the Y chromosome, it can be used to determine the sex of organisms for which a reference genome was obtained from a female or for which only the X chromosome is reported. Such taxa include the domestic cat, sheep, goat, and horse; and non-domesticated animals such as the Sumatran orangutan, western lowland gorilla and meerkat.

This study investigated the use of sexually dimorphic metrics of the first permanent maxillary molar (M1 ) to determine sex in adult and immature individuals within and between populations.
Ten M1 dimensions were measured in 91 adults (19-55 years) and 58 immatures (5-18 years) from two English populations, one of documented sex (Spitalfields crypt) and another of morphologically-assigned sex (Black Gate). Preliminary statistical analysis was undertaken to explore bilateral differences and variation by age and sex, followed by multivariate analyses to predict sex from dental metrics.
Both cross-validated linear discriminant analysis and binary logistic regression predicted biological sex consistent with known sex in 94.6% of adults and 90.9% of immatures. When functions extracted from the Spitalfields data were used to assign sex to Black Gate adults, consistency with morphological sex varied from 83.3% to 57.7%. A new function developed on Black Gate resulted in only a 4.8% increase in maximum accuracy but reduced bias. The immature cohort comprised 19 (52.8%) males and 17 (47.2%) females.
This study demonstrates substantial sexual dimorphism in a single tooth which is commonly preserved in archaeological and forensic contexts. It successfully assigns biological sex to immatures from 5 years of age with substantially greater accuracy than any other morphological or metric method. We suggest that accurate cross-population functions based on dentition require a trade-off between accuracy and applicability, and that functions extracted from populations of documented sex can be used to assign sex to other archaeological and forensic remains.

Sex determination of unknown individuals is one of the primary goals of Physical and Forensic Anthropology. The adult skeleton can be sexed using both morphological and metric traits on a large number of bones. The human pelvis is often used as an important element of adult sex determination. However, studies carried out about the pelvic bone in subadult individuals present several limitations due the absence of sexually dimorphic characteristics. In this study, we analyse the sexual dimorphism of the immature pubis and ischium bones, attending to their shape (Procrustes residuals) and size (centroid size), using an identified sample of subadult individuals composed of 58 individuals for the pubis and 83 for the ischium, aged between birth and 1year of life, from the Granada osteological collection of identified infants (Granada, Spain). Geometric morphometric methods and discriminant analysis were applied to this study. The results of intra- and inter-observer error showed good and excellent agreement in the location of coordinates of landmarks and semilandmarks, respectively. Principal component analysis performed on shape and size variables showed superposition of the two sexes, suggesting a low degree of sexual dimorphism. Canonical variable analysis did not show significant changes between the male and female shapes. As a consequence, discriminant analysis with leave-one-out cross validation provided low classification accuracy. The results suggested a low degree of sexual dimorphism supported by significant sexual dimorphism in the subadult sample and poor cross-validated classification accuracy. The inclusion of centroid size as a discriminant variable does not imply a significant improvement in the results of the analysis. The similarities found between the sexes prevent consideration of pubic and ischial morphology as a sex estimator in early stages of development. The authors suggest extending this study by analysing the different trajectories of shape and size in later ontogeny between males and females.

Although the presence of a preauricular sulcus is often cited as an indicator of female sex, very little research has been conducted to substantiate this claim. This article examines both the relationship between the incidence and morphology of the preauricular sulcus and sex.
A total of 500 left ossa coxae from the Hamann-Todd Human Osteological Collection belonging to 261 males and 239 females were examined to investigate the relationship between the preauricular sulcus and sex. The sample includes both American Blacks and Whites. Each individual was scored based on groove presence and morphology. Differences between the sexes in terms of overall prevalence and morphology were investigated using chi-squared tests. Additionally, possible relationships between ancestry and the manifestation of the preauricular sulcus were investigated using multiple regression analysis.
Overall, the presence or absence of the preauricular sulcus resulted in correct sex assessment in 75.8% of ossa coxae. However, the accuracy differed between the sexes, with 62.84% of males and 89.96% of females being assessed correctly. Males were significantly more likely to display a short, narrow sulcus (score 4) than females, whereas females were observed to be significantly more likely to have a wide, long sulcus. Ancestry was not related to sulcus manifestation.
The results presented here suggest that the presence of a preauricular sulcus should not be considered positive proof of female sex. However, the complete lack of a sulcus could be considered suggestive of male sex.

Sexual dimorphism is a crucial characteristic of skeleton. In the last years, volumetric and surface 3D acquisition systems have enabled anthropologists to assess surfaces and volumes, whose potential still needs to be verified. This article aimed at assessing volume and linear parameters of the first metatarsal bone through 3D acquisition by laser scanning. Sixty-eight skeletons underwent 3D scan through laser scanner: Seven linear measurements and volume from each bone were assessed. A cutoff value of 13,370 mm3 was found, with an accuracy of 80.8%. Linear measurements outperformed volume: metatarsal length and mediolateral width of base showed higher cross-validated accuracies (respectively, 82.1% and 79.1%, raising at 83.6% when both of them were included). Further studies are needed to verify the real advantage for sex assessment provided by volume measurements.