Spatial analysis of the ancient proteome of archeological teeth using mass spectrometry imaging.

RATIONALE: Proteins extracted from archaeological bone and teeth are utilised for investigating the phylogeny of extinct and extant species, the biological sex and age of past individuals, as well as ancient health and physiology. However, variable preservation of proteins in archaeological materials represents a major challenge. METHODS: To better understand the spatial distribution of ancient proteins preserved within teeth, we applied matrix assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) for the first time to bioarchaeological samples to visualise the intensity of proteins in archaeological teeth thin sections. We specifically explored the spatial distribution of four proteins (collagen type I, of which the chains alpha-1 and alpha-2, alpha-2-HS-glycoprotein, haemoglobin subunit alpha and myosin light polypeptide 6). RESULTS: We successfully identified ancient proteins in archaeological teeth thin sections using mass spectrometry imaging. The data are available via ProteomeXchange with identifier PXD038114. However, we observed that peptides did not always follow our hypotheses for their spatial distribution, with distinct differences observed in the spatial distribution of several proteins, and occasionally between peptides of the same protein. CONCLUSIONS: While it remains unclear what causes these differences in protein intensity distribution within teeth, as revealed by MALDI-MSI in this study, we have demonstrated that MALDI-MSI can be successfully applied to mineralised bioarchaeological tissues to detect ancient peptides. In future applications, this technique could be particularly fruitful not just for understanding the preservation of proteins in a range of archaeological materials, but making informed decisions on sampling strategies and the targeting of key proteins of archaeological and biological interest.

Elucidating inequality in Nubia: an examination of entheseal changes at Kerma (Sudan).

Located 10 km south of the Third Cataract of the Nile River, the ancient city of Kerma was once capital to the second largest state in Africa. The Eastern Cemetery at Kerma (∼4 km east of city center) encompasses 80+ hectares and was used over a period of 1,500 years (3,200-1,500 BC). Excavated in the early 20th century by George Reisner, the cemetery contained an estimated 20,000-40,000 individuals. Reisner classified these burials into multiple categories, including chiefs and human sacrifices, based on burial position and grave goods. This study investigates the skeletal embodiment of social inequality by examining variation in entheseal severity between the Kerma burial classifications. Seventeen entheses were examined using the Hawkey and Merbs (1995) scoring method (n = 205 individuals); age, sex, and body size variables were considered by employing Mann-Whitney U tests and partial Spearman's correlations. This analysis suggests that significant differences in entheseal changes existed between select burial types. Specifically, "corridor sacrifices" had significantly higher rates of entheseal changes while "chiefs" and "subsidiary burials" had similar entheseal changes; furthermore, within these burial categories, males had higher entheseal scores despite body size controls. The elevated entheseal changes in the sacrificial burials may be due to an intensive agro-pastoral lifestyle or other demanding forms of manual labor. In conclusion, the disparity of entheseal markers between burial subgroups at Kerma might reflect a degree of social inequality within this state level society. This bioarchaeological research informs our understanding of socially-defined categories of persons as well as everyday life in Ancient Kerma.

Chronic Maxillary Sinusitis: A comparison of osteological and CT methods of diagnosis.

OBJECTIVE: To assess the differences between endoscopic and radiological methods of analysis for diagnosing chronic maxillary sinusitis (CMS) in archaeological skeletal remains. MATERIALS: 32 crania from a Dutch post-medieval rural population. METHODS: We assessed the presence of bone changes indicative of CMS (i.e., bone growth and bone resorption) both endoscopically and through computed tomography (CT), and then compared results. RESULTS: We observed moderate agreement between bone growth scores obtained through endoscopy and CT, and fair agreement when assessing bone resorption. CONCLUSIONS: CMS prevalence rates observed through CT may be comparable to rates assessed endoscopically, although caution is needed when making direct comparisons. SIGNIFICANCE: This is the first study comparing data obtained through endoscopy and radiological methods in the study of CMS, informing paleopathologists about potential biases in data comparison. LIMITATIONS: Our small sample size likely impacted results. SUGGESTIONS FOR FURTHER RESEARCH: Further research is advised to fully explore the comparability of endoscopic and radiological method of analysis in the study of sinusitis. DATA AVAILABILITY: The datasets generated and analyzed during the current study are available upon request.

An Accommodation for Whom? Has the COVID-19 Pandemic Changed the Landscape of Flexible and Remote Work for Workers with Disabilities?

At the start of the COVID-19 pandemic, flexible and remote work was viewed as a silver bullet that would increase employment rates among people with disabilities. This view fails to recognize that not all workers with disabilities can obtain jobs that can be done remotely or on a flexible schedule. Data from the 2019 and 2021 years of the Current Population Survey and the American Community Survey were used to examine if disabled workers' gender, race, ethnicity, age, and education, increase (or decrease) their chances of accessing flexible and remote work and if the group of workers with disabilities who access such options expanded since the COVID-19 pandemic. Findings indicate that compared to their non-disabled counterparts, prior to the pandemic, workers with disabilities reported similar rates of flexible and remote work. Workers with disabilities, however, had lower rates of remote work after the start of the pandemic. Regardless of year, flexible and remote work rates vary by demographic group, with disabled workers who are white, female, and college-educated more likely to access these options than multiply marginalized disabled workers.

Reframing antimicrobial resistance as a continuous spectrum of manifestations.

To fight antimicrobial resistance (AMR), we must recognize and target all its manifestations. In this review, we briefly summarize the history that led to recognition of the various manifestations of AMR in bacterial pathogens and the ways in which they interrelate. We emphasize the importance of distinguishing between AMR arising from genetic alterations versus induction of endogenous machinery in response to environmental triggers, including - paradoxically - stresses from host immunity and antimicrobial therapy. We present an integrated view of AMR by reframing it as a spectrum of phenotypes within a continuous three-dimensional space defined by the growth rate, prevalence, and kill rate of cells displaying AMR. Finally, we reflect on strategies that may help stem the emergence of AMR.

A distant city: Assessing the impact of Dutch socioeconomic developments on urban and rural health using respiratory disease as a proxy.

OBJECTIVES: To investigate the prevalence of respiratory disease in several populations from the Netherlands across different time periods and socioeconomic conditions. MATERIALS: We analyzed 695 adult individuals from six different Dutch contexts of urban and rural settlements dating to different time periods (i.e., early-medieval, late-medieval, post-medieval). METHODS: For each individual, the presence/absence of chronic maxillary sinusitis, otitis media, and inflammatory periosteal reaction on ribs was recorded macroscopically according to accepted methods. RESULTS: Statistically significant associations were found in the presence of sinusitis diachronically (early-medieval to late-medieval period, and early-medieval to post-medieval period) both in rural and urban environments. Differences in prevalence rates of otitis media were found statistically significant when comparing rural to urban environments in the early-medieval and late-medieval periods. CONCLUSION: Our results suggest that factors such as increased contact between towns and countryside, higher population densities, and intensification of agricultural production impacted the respiratory health of past populations both in rural and urban settings. SIGNIFICANCE: Our study provides new insights into the impact of environmental changes and urbanization on respiratory disease prevalence, shedding light on the relationship between health and changing social and environmental contexts. LIMITATIONS: Research limitations included the complex etiology of respiratory diseases, and the impact of uncontrollable factors such as hidden heterogeneity, selective mortality, and rural-to-urban migration. FUTURE RESEARCH: Further research in different contexts is advised in order to continue exploring urbanization and its impact on human health across both time and space.

Osteological, multi-isotope and proteomic analysis of poorly-preserved human remains from a Dutch East India Company burial ground in South Africa.

Skeletal remains discovered in Simon's Town, South Africa, were hypothesised as being associated with a former Dutch East India Company (VOC) hospital. We report a novel combined osteological and biochemical approach to these poorly-preserved remains. A combined strontium (87Sr/86Sr), oxygen (δ18OVPDB) and carbon (δ13CVPDB) isotope analysis informed possible childhood origins and diet, while sex-specific amelogenin enamel peptides revealed biological sex. Osteological analyses presented evidence of residual rickets, a healed trauma, dental pathological conditions, and pipe notches. The combined isotope analyses yielded results for 43 individuals which suggested a diverse range of geological origins, including at least 16% of the population being non-local. The inclusion of δ13CVPDB had intriguing implications for three individuals who likely did not have origins in the Cape Town region nor in Europe. Peptide analysis on the dental enamel of 25 tested individuals confirmed they were all biologically male. We suggest that isolated enamel may provide crucial information about individuals' pathological conditions, geographical origins, diet, and biological sex. These data further demonstrated that a combined approach using multiple osteological and biochemical methods is advantageous for human remains which are poorly preserved and can contextualise a site with little direct evidence.

Antibody-based sex determination of human skeletal remains.

Assignment of biological sex to skeletal remains is critical in the accurate reconstruction of the past. Analysis of sex-chromosome encoded AMELX and AMELY peptides from the enamel protein amelogenin underpins a minimally destructive mass spectrometry (MS) method for sex determination of human remains. However, access to such specialist approaches limits applicability. As a convenient alternative, we generated antibodies that distinguish human AMELX and AMELY. Purified antibodies demonstrated high selectivity and quantitative detection against synthetic peptides by ELISA. Using acid etches of enamel from post-medieval skeletons, antibody determinations corrected osteological uncertainties and matched parallel MS, and for Bronze Age samples where only enamel was preserved, also matched MS analyses. Toward improved throughput, automated stations were applied to analyze 19th-century teeth where sex of individuals was documented, confirming MS can be bypassed. Our immunological tools should underpin development of routine, economical, high-throughput methods for sex determination, potentially even in a field setting.

Activity patterns in New Kingdom Nubia: an examination of entheseal remodeling and osteoarthritis at Tombos.

The effects of Egyptian imperial expansion into Nubia during the New Kingdom Period (1,550-1,069 BC) have been debated. Here, the impacts of the Egyptian Empire are investigated through an examination of osteological indicators of activity at the archaeological site of Tombos. Entheseal changes to fibrocartilaginous attachment sites and osteoarthritis are examined to infer what types of physical activities this colonial town was engaging in. Many of the skeletal remains at Tombos were commingled due to looting in antiquity; undisturbed burials are presented as a subsample of the population (n = 28) in which age, sex, and body size can be considered. The total sample (n = 85) is then analyzed to better understand overall levels of activity. A number of Nile River Valley bioarchaeological samples are used as points of comparison to the Tombos population. Results indicate that the inhabitants of Tombos had relatively low entheseal remodeling scores; this is highlighted when Tombos is juxtaposed with comparative samples, particularly in men. Furthermore, osteoarthritis, as assessed by eburnation, was also markedly infrequent at Tombos. Collectively, these results indicate a relatively low level of activity and support the hypothesis that Tombos may have served as an administrative center.

Hemoglobin, growth, and attention of infants in southern Ethiopia.

Male and female infants from rural Ethiopia were tested to investigate relations among hemoglobin (Hb), anthropometry, and attention. A longitudinal design was used to examine differences in attention performance from 6 (M = 24.9 weeks, n = 89) to 9 months of age (M = 40.6 weeks, n = 85), differences hypothesized to be related to changes in iron status and growth delays. Stunting (length-for-age z scores < -2.0) and attention performance, t(30) = -2.42, p = .022, worsened over time. Growth and Hb predicted attention at 9 months, R(2) = .15, p < .05, but not at 6. The study contributes to the knowledge base concerning the relations among Hb, early growth, and attention.

Multiform antimicrobial resistance from a metabolic mutation.

A critical challenge for microbiology and medicine is how to cure infections by bacteria that survive antibiotic treatment by persistence or tolerance. Seeking mechanisms behind such high survival, we developed a forward-genetic method for efficient isolation of high-survival mutants in any culturable bacterial species. We found that perturbation of an essential biosynthetic pathway (arginine biosynthesis) in a mycobacterium generated three distinct forms of resistance to diverse antibiotics, each mediated by induction of WhiB7: high persistence and tolerance to kanamycin, high survival upon exposure to rifampicin, and minimum inhibitory concentration-shifted resistance to clarithromycin. As little as one base change in a gene that encodes, a metabolic pathway component conferred multiple forms of resistance to multiple antibiotics with different targets. This extraordinary resilience may help explain how substerilizing exposure to one antibiotic in a regimen can induce resistance to others and invites development of drugs targeting the mediator of multiform resistance, WhiB7.

Age and disability employment discrimination: occupational rehabilitation implications.

INTRODUCTION: As concerns grow that a thinning labor force due to retirement will lead to worker shortages, it becomes critical to support positive employment outcomes of groups who have been underutilized, specifically older workers and workers with disabilities. Better understanding perceived age and disability discrimination and their intersection can help rehabilitation specialists and employers address challenges expected as a result of the evolving workforce. METHODS: Using U.S. Equal Employment Opportunity Commission Integrated Mission System data, we investigate the nature of employment discrimination charges that cite the Americans with Disabilities Act or Age Discrimination in Employment Act individually or jointly. We focus on trends in joint filings over time and across categories of age, types of disabilities, and alleged discriminatory behavior. RESULTS: We find that employment discrimination claims that originate from older or disabled workers are concentrated within a subset of issues that include reasonable accommodation, retaliation, and termination. Age-related disabilities are more frequently referenced in joint cases than in the overall pool of ADA filings, while the psychiatric disorders are less often referenced in joint cases. When examining charges made by those protected under both the ADA and ADEA, results from a logit model indicate that in comparison to charges filed under the ADA alone, jointly-filed ADA/ADEA charges are more likely to be filed by older individuals, by those who perceive discrimination in hiring and termination, and to originate from within the smallest firms. CONCLUSION: In light of these findings, rehabilitation and workplace practices to maximize the hiring and retention of older workers and those with disabilities are discussed.

Biology of antimicrobial resistance and approaches to combat it.

Insufficient development of new antibiotics and the rising resistance of bacteria to those that we have are putting the world at risk of losing the most widely curative class of medicines currently available. Preventing deaths from antimicrobial resistance (AMR) will require exploiting emerging knowledge not only about genetic AMR conferred by horizontal gene transfer or de novo mutations but also about phenotypic AMR, which lacks a stably heritable basis. This Review summarizes recent advances and continuing limitations in our understanding of AMR and suggests approaches for combating its clinical consequences, including identification of previously unexploited bacterial targets, new antimicrobial compounds, and improved combination drug regimens.

Getting Rid of Dichotomous Sex Estimations: Why Logistic Regression Should be Preferred Over Discriminant Function Analysis.

Sex estimation is an important part of creating a biological profile for skeletal remains in forensics. The commonly used methods for developing sex estimation equations are discriminant function analysis (DFA) and logistic regression (LogR). LogR equations provide a probability of the predicted sex, while DFA relies on cutoff points to segregate males and females, resulting in a rigid dichotomization of the sexes. This is problematic because sexual dimorphism exists along a continuum and there can be considerable overlap in trait expression between the sexes. In this study, we used humeral measurements to compare the performance of DFA and LogR and found them to be very similar under multiple conditions. The overall cross-validated (leave-one-out) accuracy of DFA (75.76-95.14%) was slightly higher than LogR (75.76-93.82%) for simple and multiple variable equations, and also performed better under varying sample sizes (94.03% vs. 93.78%). Three of five DFA equations outperformed LogR under the B index, while all five LogR equations outperformed the DFA equations under the Q index. Both methods saw an improvement in overall accuracy (DFA: 86.74-95.79%; LogR: 86.74-95.76%) when individuals with a classification probability lower than 0.80 were excluded. Additionally, we propose a method for calculating additional cutoff points (PMarks) based on posterior probability values. In conclusion, we recommend using LogR over DFA due to the increased flexibility, robusticity, and benefits for future users of the statistical models; however, if DFA is preferred, use of the proposed PMarks facilitates future analysis while avoiding unnecessary dichotomization.

Primary care physician specialty referral decision making: patient, physician, and health care system determinants.

PURPOSE: To examine the effects of patient, physician, and health care system characteristics on primary care physicians' (PCPs') specialty referral decision making. METHODS: Physicians (n=142) and their practices (n=83) located in 30 states completed background questionnaires and collected survey data for all patient visits (n=34,069) made during 15 consecutive workdays. The authors modeled the occurrence of any specialty referral, which occurred during 5.2% of visits, as a function of patient, physician, and health care system structural characteristics. A subanalysis was done to examine determinants of referrals made for discretionary indications (17% of referrals), operationalized as problems commonly managed by PCPs, high level of diagnostic and therapeutic certainty, low urgency for specialist involvement, and cognitive assistance only requested from the specialist. RESULTS: Patient characteristics had the largest effects in the any-referral model. Other variables associated with an increased risk of referral included PCPs with less tolerance of uncertainty, larger practice size, health plans with gate-keeping arrangements, and practices with high levels of managed care. The risk of a referral being made for discretionary reasons was increased by capitated primary care payment, internal medicine specialty of the PCP, high concentration of specialists in the community, and higher levels of managed care in the practice. CONCLUSIONS: PCPs' referral decisions are influenced by a complex mix of patient, physician, and health care system structural characteristics. Factors associated with more discretionary referrals may lower PCPs' thresholds for referring problems that could have been managed in their entirety within primary care settings.

Trends and Patterns in Age Discrimination in Employment Act (ADEA) Charges.

The Age Discrimination in Employment Act (ADEA) protects individuals aged 40 years and over from discrimination throughout the employment process. Using data on ADEA charges from the Equal Employment Opportunity Commission from 1993 to 2010, we present labor force-adjusted charge rates demonstrating that the highest charge rates are among those in the preretirement age range, and only the rate of charges among those aged 65 years and older has not decreased. We examine characteristics of ADEA charges including the prevalence of different alleged discriminatory actions (or issues) and highlight the increasing proportion of age discrimination charges that are jointly filed with other antidiscrimination statutes. Through a regression analysis, we find that the likelihood of citing various issues differs by charging party characteristics, such as age, gender, and minority status, and on charges that cite only age discrimination as compared to those that are jointly filed. Implications of these findings for employers are discussed.

Family physicians' referral decisions: results from the ASPN referral study.

OBJECTIVE: To examine family physicians' referral decisions, which we conceptualized as having 2 phases: whether to refer followed by to whom to refer. STUDY DESIGN: Prospective cohort study. POPULATION: All visits (N = 34,519) and new referrals (N = 2534) occurring during 15 consecutive business days in the offices of 141 family physicians in 87 practices located in 31 states. OUTCOMES MEASURED: Rates of referral, reasons for referral, practitioners referred to, health problems prompting referral, and reasons for selecting particular specialists. RESULTS: Approximately 1 in 20 (5.1%) office visits led to referral. Although 68% of referrals were made by physicians during office visits, 18% were made by physicians during telephone conversations with patients, 11% by office staff with input from the physician, and 3% by staff without physician input. Physicians endorsed a mean of 1.8 reasons for making a referral. They sought specialists' advice on either diagnosis or treatment for 52.1% of referrals and asked the specialist to direct medical management for 25.9% and surgical management for 37.8%. Patient request was one reason for 13.6% of referrals. Fifty conditions accounted for 76% of all referrals. Surgical specialists were sent the largest share of referrals (45.4%), followed by medical specialists (31.0%), nonphysician clinicians (12.1%), obstetrician-gynecologists (4.6%), mental health professionals (4.2%), other practitioners (2.0%), and generalists (0.8%). Physicians recommended a specific practitioner to the patient for most (86.2%) referrals. Personal knowledge of the specialist was the most important reason for selecting a specific specialist. CONCLUSIONS: Referrals are commonly made during encounters other than office visits, such as telephone conversations or staff-patient interactions, in primary care practice. Training in the referral process should ensure that family physicians obtain the skills necessary to expand their scope of practice, when appropriate; determine when and why a patient should be referred; and identify the type of practitioner to whom the patient should be sent.

Variability in physician referral decisions.

BACKGROUND: Because it is possible that part of the variability in frequency of interventions and even in outcomes results from the variability in referral patterns of primary care physicians, our objectives were to examine primary care physician decision making about referrals for several common adult and childhood conditions. METHODS: One hundred thirty-six family physicians in 80 office-based practices recorded reasons for referral to the most common types of specialists and the expectation of the specialist for conditions with at least seven referrals to each specialist. RESULTS: By far the most referrals were expected to be short-term (< 12 months); for more than 50%, the referrals were for consultation only (rather than direct intervention). For most of the 10 types of conditions, there were no apparent differences in reason for or expectation of the referral that would explain the choice of different types of specialists for referral. Expectations for shared care were generally more common in referrals to nonphysicians than to physicians. CONCLUSIONS: This study revealed unexplained variability among family physicians in the specialists to whom patients are referred for specific conditions. Why some patients with the same condition are referred to surgeons and others to medical specialists is unclear, at least in the context of expectations for referral as being long-term vs short-term or consultative vs referral for definitive management. The impact of this variability on costs and outcome could be considerable and deserves more intensive study.

Vulnerability and the patient-practitioner relationship: the roles of gatekeeping and primary care performance.

OBJECTIVES: We examined whether patients' perceptions of their relationships with primary care practitioners (PCPs) vary by vulnerability status and assessed the extent to which gatekeeping arrangements and primary care performance moderate potential disparities. METHODS: We used the nationally representative 1996-1997 Community Tracking Study Household Survey as our data source. RESULTS: Whites reported better patient-practitioner relationships than minorities. Requirements that patients select a PCP and obtain referral authorization neither reduced nor exacerbated racial disparities in the patient-practitioner relationship. On the other hand, access to and continuity with a PCP substantively reduced disparities, especially for the most vulnerable group. CONCLUSIONS: Enhancing primary care performance may reduce some of the barriers to care experienced by vulnerable populations, thereby improving patients' relationships with their PCPs.