Amelogenin genes are located on both X and Y sex chromosomes in humans and are a major focus of DNA-based sex estimation methods. Amelogenin proteins, AMELX_HUMAN and AMELY_HUMAN, are expressed in the tooth organ and play a major role in mineralization of enamel, the most taphonomically resistant, archaeologically persistent human tissue. We describe shotgun liquid chromatography mass spectrometry analysis of 40 enamel samples representing 25 individuals, including modern third molars and archaeological teeth from open-air contexts including permanent adult (400-7300 BP) and deciduous teeth (100-1000 BP). Peptides specific to the X-chromosome isoform of amelogenin were detected in all samples. Peptides specific to the sexually dimorphic Y-chromosome isoform were also detected in 26 samples from 13 individuals, across all time periods, including previously unsexed deciduous teeth from archaeological contexts. While the signal of each gene product can vary by more than an order of magnitude, we show close agreement between osteological and amelogenin-based sex estimation and thus demonstrate that the protein-based signal can reliably be obtained from open-air archaeological contexts dating to at least 7300 years ago. While samples with AMELY_HUMAN peptides are unambiguously male, samples with no AMELY_HUMAN signal may either be low signal male false negative samples or female samples. In order to estimate sex in these samples we developed a probability curve of female sex as a function of the logarithm of AMELX_HUMAN signal (p < 0.0001) using logistic regression. This is also the first demonstration using proteomics to estimate sex in deciduous teeth and pushes back the application of the method to teeth that are at least 7300 years old.

Sex estimation of skeletons is fundamental to many archaeological studies. currently, three approaches are available to estimate sex-osteology, genomics, or proteomics, but little is known about the relative reliability of these methods in applied settings. We present matching osteological, shotgun-genomic, and proteomic data to estimate the sex of 55 individuals, each with an independent radiocarbon date between 2,440 and 100 cal BP, from two ancestral Ohlone sites in Central California. Sex estimation was possible in 100% of this burial sample using proteomics, in 91% using genomics, and in 51% using osteology. Agreement between the methods was high, however conflicts did occur. Genomic sex estimates were 100% consistent with proteomic and osteological estimates when DNA reads were above 100,000 total sequences. However, more than half the samples had DNA read numbers below this threshold, producing high rates of conflict with osteological and proteomic data where nine out of twenty conditional DNA sex estimates conflicted with proteomics. While the DNA signal decreased by an order of magnitude in the older burial samples, there was no decrease in proteomic signal. We conclude that proteomics provides an important complement to osteological and shotgun-genomic sex estimation. Biological sex plays an important role in the human experience, correlating to lifespan, reproduction, and a wide range of other biological factors 1-5. Sex and gender are also fundamental in structuring an array of cultural behaviors, including residence patterns, kinship, economic roles, and identity construction and expression 6-9. How sex interacts with gender and these particular issues is not static and can vary in detail across societies and over time 10-12. It is not surprising that sex is one of the most basic and important measures in bioarchaeological and forensic analyses. Typically, osteological features are used to estimate sex of skeletal remains, and the most widely used marker is the morphology of the os coxae 13-16. However, appropriate markers are not always sufficiently expressed or preserved to estimate sex using morphological criteria 17. A lack of sexually-dimorphic markers is especially acute for skeletons of infants and children who have not undergone puberty. Mortuary practices, such as cremation or secondary burial in charnel houses, can also can impose limitations on the utility of osteological sex estimates 18. The advent of DNA sequencing made it possible to use skeletal remains to estimate the sex of very young individuals; it also expanded sex estimations for fragmentary, pathological, and degraded skeletal materials 19-21. More recently, development of massively parallel DNA sequencing greatly improved genome coverage in archaeological samples 22-25. In addition to providing detailed genetic information, this allows biological sex to be estimated from shotgun sequencing data 25-27. These approaches were an improvement over earlier PCR-based marker

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PURPOSE Cervical cancer remains the eighth leading cause of cancer mortality among women in Taiwan. Despite availability of a National Health Insurance program with free yearly screenings, Papanicolaou testing and human papillomavirus (HPV) vaccination rates have been historically low in Taiwan. Even in the United States, cervical cancer screening rates for Asian women are significantly lower than other ethnic groups. The goal is to direct future interventions by providing insight on barriers leading to low screening and vaccination rates among Taiwanese and Asian women. METHODS Anonymous surveys without patient identifiers were randomly administered to patients at a traditional Chinese medicine clinic in Hualien City in June 2018. Inclusion criterion was females. No exclusion criteria were defined. Participants provided written consent. Sixty-three completed surveys were received. A χ2 test was used to determine statistical significance (α = 0.05). RESULTS Formal education level correlated with increased knowledge of HPV ( P = .001), its route of transmission ( P = .044), its link to genital cancer ( P = .0024), and HPV vaccination ( P = .0039). Women were more likely to have Papanicolaou smears if they were older than 30 years of age ( P = .0033), visited the gynecologist ( P < .001), or were recommended one by their physicians ( P < .001). Although 57% of respondents knew of the HPV vaccine, only 19% were vaccinated. Among those not vaccinated, most cited reasons included an inability to find a physician providing it (23.5%), safety concerns (16.4%), belief that it encourages sexual behavior (14.5%), and high out-of-pocket expense (9.1%). CONCLUSION Knowledge of HPV does not predict a higher adherence to cervical screening guidelines. Instead, diligent physician recommendations on Papanicolaou smears can elevate adherence rates among patients. Significant contributors to low HPV vaccination rates in rural Taiwan include lack of awareness and access to the vaccine. Our study emphasizes the physician-patient relationship as a means to target vulnerable populations and increase rates of cervical cancer screening and HPV vaccination.