Applying an evolutionary mismatch framework to understand disease susceptibility.

Noncommunicable diseases (NCDs) are on the rise worldwide. Obesity, cardiovascular disease, and type 2 diabetes are among a long list of "lifestyle" diseases that were rare throughout human history but are now common. The evolutionary mismatch hypothesis posits that humans evolved in environments that radically differ from those we currently experience; consequently, traits that were once advantageous may now be "mismatched" and disease causing. At the genetic level, this hypothesis predicts that loci with a history of selection will exhibit "genotype by environment" (GxE) interactions, with different health effects in "ancestral" versus "modern" environments. To identify such loci, we advocate for combining genomic tools in partnership with subsistence-level groups experiencing rapid lifestyle change. In these populations, comparisons of individuals falling on opposite extremes of the "matched" to "mismatched" spectrum are uniquely possible. More broadly, the work we propose will inform our understanding of environmental and genetic risk factors for NCDs across diverse ancestries and cultures.

Variation in diurnal cortisol patterns among the Indigenous Shuar of Amazonian Ecuador.

OBJECTIVES: The hypothalamic-pituitary-adrenal (HPA) axis and its primary end product, the glucocorticoid cortisol, are major components of the evolved human stress response. However, most studies have examined these systems among populations in high-income settings, which differ from the high pathogen and limited resource contexts in which the HPA axis functioned for most of human evolution. METHODS: We investigated variability in diurnal salivary cortisol patterns among 298 Indigenous Shuar from Amazonian Ecuador (147 males, 151 females; age 2-86 years), focusing on the effects of age, biological sex, and body mass index (BMI) in shaping differences in diurnal cortisol production. Saliva samples were collected three times daily (waking, 30 minutes post-waking, evening) for three consecutive days to measure key cortisol parameters: levels at waking, the cortisol awakening response, the diurnal slope, and total daily output. RESULTS: Age was positively associated with waking levels and total daily output, with Shuar juveniles and adolescents displaying significantly lower levels than adults (p < .05). Sex was not a significant predictor of cortisol levels (p > .05), as Shuar males and females displayed similar patterns of diurnal cortisol production across the life course. Moreover, age, sex, and BMI significantly interacted to moderate the rate of diurnal cortisol decline (p = .027). Overall, Shuar demonstrated relatively lower cortisol concentrations than high-income populations. CONCLUSIONS: This study expands the documented range of global variation in HPA axis activity and diurnal cortisol production and provides important insights into the plasticity of human stress physiology across diverse developmental and socioecological settings.

Pathogen disgust sensitivity protects against infection in a high pathogen environment.

Disgust is hypothesized to be an evolved emotion that functions to regulate the avoidance of pathogen-related stimuli and behaviors. Individuals with higher pathogen disgust sensitivity (PDS) are predicted to be exposed to and thus infected by fewer pathogens, though no studies have tested this directly. Furthermore, PDS is hypothesized to be locally calibrated to the types of pathogens normally encountered and the fitness-related costs and benefits of infection and avoidance. Market integration (the degree of production for and consumption from market-based economies) influences the relative costs/benefits of pathogen exposure and avoidance through sanitation, hygiene, and lifestyle changes, and is thus predicted to affect PDS. Here, we examine the function of PDS in disease avoidance, its environmental calibration, and its socioecological variation by examining associations among PDS, market-related lifestyle factors, and measures of bacterial, viral, and macroparasitic infection at the individual, household, and community levels. Data were collected among 75 participants (ages 5 to 59 y) from 28 households in three Ecuadorian Shuar communities characterized by subsistence-based lifestyles and high pathogen burden, but experiencing rapid market integration. As predicted, we found strong negative associations between PDS and biomarkers of immune response to viral/bacterial infection, and weaker associations between PDS and measures of macroparasite infection, apparently mediated by market integration-related differences. We provide support for the previously untested hypothesis that PDS is negatively associated with infection, and document variation in PDS indicative of calibration to local socioeconomic conditions. More broadly, findings highlight the importance of evolved psychological mechanisms in human health outcomes.

Dual burden of infectious and chronic disease in low-resource U.S. communities: examining relationships between infection, adiposity, and inflammation.

BACKGROUND: Rising global obesity rates are linked with inflammation and associated morbidities. These negative outcomes are generally more common in low-resource communities within high-income countries; however, it is unclear how frequent infectious disease exposures in these settings may influence the relationship between adiposity and inflammation. AIM: We test associations between adiposity measures and distinct forms of inflammation among adults (n = 80) living in low-resource U.S. communities experiencing high levels of obesity and pathogen exposure. SUBJECTS AND METHODS: Adiposity measures included BMI and percent body fat. Inflammation measures included systemic inflammation (C-reactive protein [CRP]) and localised intestinal inflammation (faecal calprotectin [FC]). The relationship between a condition characterised by elevated inflammation (Helicobacter pylori infection) and adiposity was also considered. RESULTS: Adiposity was not significantly related to FC concentration. However, both adiposity measures were positively related with odds of CRP elevation and H. pylori infection was associated with significantly lower adiposity measures (all p < 0.05). CONCLUSION: For this disadvantaged U.S. sample, the association between adiposity and inflammation varies by the systemic/localised nature of inflammation and the likely underlying cause of inflammation. Defining these associations will improve understanding of how rising obesity rates shape long-term health inequities, with implications for more effective intervention design.

Minimally invasive biomarkers in human and non-human primate evolutionary biology: Tools for understanding variation and adaptation.

BACKGROUND: The use of minimally invasive biomarkers (MIBs - physiological biomarkers obtained from minimally invasive sample types) has expanded rapidly in science and medicine over the past several decades. The MIB approach is a methodological strength in the field of human and non-human primate evolutionary biology (HEB). Among humans and our closest relatives, MIBs provide unique opportunities to document phenotypic variation and to operationalize evolutionary hypotheses. AIMS: This paper overviews the use of MIBs in HEB. Our objectives are to (1) highlight key research topics which successfully implement MIBs, (2) identify promising yet under-investigated areas of MIB application, and (3) discuss current challenges in MIB research, with suggestions for advancing the field. DISCUSSION AND CONCLUSIONS: A range of MIBs are used to investigate focal topics in HEB, including energetics and life history variation/evolution, developmental plasticity, and social status and dominance relationships. Nonetheless, we identify gaps in existing MIB research on traits such as physical growth and gut function that are central to the field. Several challenges remain for HEB research using MIBs, including the need for additional biomarkers and methods of assessment, robust validations, and approaches that are standardized across labs and research groups. Importantly, researchers must provide better support for adaptation and fitness effects in hypothesis testing (e.g., by obtaining complementary measures of energy expenditure, demonstrating redundancy of function, and performing lifetime/longitudinal analyses). We point to continued progress in the use of MIBs in HEB to better understand the past, present, and future of humans and our closest primate relatives.

Age-related patterns of cytomegalovirus antibodies accompanying Epstein-Barr virus co-infection.

OBJECTIVE: Cytomegalovirus (CMV) infection is associated with age-related chronic disease, and co-infection with Epstein-Barr virus (EBV) may compound disease risk. We aimed to assess the frequency of CMV infection and its relationship with age among EBV seropositive individuals in an Indigenous Amazonian population. METHODS: We report concentrations of CMV and EBV antibodies in dried blood spot samples collected from 157 EBV positive Shuar participants aged 15-86 years (60.5% female) to assess CMV infection rate. We used logistic and linear regression models to examine associations among CMV, EBV, and age, adjusting for sex, geographic region, and body mass index. RESULTS: Nearly two-thirds (63.1%) of EBV seropositive participants were also CMV seropositive. A 1-year increase in age was associated with 3.4% higher odds of CMV infection (OR [95% CI]: 1.034 [1.009-1.064], p = .012), but CMV antibody concentration was not significantly associated with age or EBV antibody concentration among co-infected individuals. CONCLUSIONS: Herpesvirus-related immunosenescence may be important to understanding chronic disease risk among Shuar. Future studies should further explore the role of co-infection in shaping age-related changes in immune function.

Low prevalence of anemia among Shuar communities of Amazonian Ecuador.

OBJECTIVE: Anemia is an important global health challenge. We investigate anemia prevalence among Indigenous Shuar of Ecuador to expand our understanding of population-level variation, and to test hypotheses about how anemia variation is related to age, sex, and market integration. METHODS: Hemoglobin levels were measured in a total sample of 1650 Shuar participants (ages 6 months to 86 years) from 46 communities between 2008 and 2017 to compare anemia prevalence across regions characterized by different levels of market integration. RESULTS: Shuar anemia rates among children under 15 years (12.2%), adult women (10.5%), and adult men (5.3%) were less than half of those previously documented in other neo-tropical Indigenous populations. Anemia prevalence did not vary between more traditional and market integrated communities (OR = 0.47, p = .52). However, anemia was negatively associated with body mass index (OR = 0.47, p = .002). CONCLUSIONS: Compared to other South American Indigenous populations, anemia prevalence is relatively low among Shuar of Ecuador and invariant with market integration. Understanding this pattern can provide valuable insights into anemia prevention among at-risk populations.

Childhood Daily Energy Expenditure Does Not Decrease with Market Integration and Is Not Related to Adiposity in Amazonia.

BACKGROUND: Childhood overweight and obesity (OW/OB) is increasingly centered in low- and middle-income countries (LMICs) as rural populations experience market integration and lifeway change. Most explanatory studies have relied on imprecise estimates of children's energy expenditure, restricting understanding of the relative effects of changes in diet and energy expenditure on the development of OW/OB in transitioning contexts. OBJECTIVES: This study used gold-standard measurements of children's energy expenditure to investigate the changes that underlie OW/OB and the nutrition/epidemiologic transition. METHODS: Cross-sectional data were collected from "rural" (n = 43) Shuar forager-horticulturalist children and their "peri-urban" (n = 34) Shuar counterparts (age 4-12 y) in Amazonian Ecuador. Doubly labeled water measurements of total energy expenditure (TEE; kcal/d), respirometry measurements of resting energy expenditure (REE; kcal/d), and measures of diet, physical activity, immune activity, and market integration were analyzed primarily using regression models. RESULTS: Peri-urban children had higher body fat percentage (+8.1%, P < 0.001), greater consumption of market-acquired foods (multiple P < 0.001), lower concentrations of immune activity biomarkers (multiple P < 0.05), and lower REE (-108 kcal/d, P = 0.002) than rural children. Despite these differences, peri-urban children's TEE was indistinguishable from that of rural children (P = 0.499). Moreover, although sample-wide IgG concentrations and household incomes predicted REE (both P < 0.05), no examined household, immune activity, or physical activity measures were related to children's overall TEE (all P > 0.09). Diet and energy expenditure associations with adiposity demonstrate that only reported consumption of market-acquired "protein" and "carbohydrate" foods predicted children's body fat levels (multiple P < 0.05). CONCLUSIONS: Despite underlying patterns in REE, Shuar children's TEE is not reliably related to market integration and-unlike dietary measures-does not predict adiposity. These findings suggest a leading role of changing dietary intake in transitions to OW/OB in LMICs.

Tradeoffs between immune function and childhood growth among Amazonian forager-horticulturalists.

Immune function is an energetically costly physiological activity that potentially diverts calories away from less immediately essential life tasks. Among developing organisms, the allocation of energy toward immune function may lead to tradeoffs with physical growth, particularly in high-pathogen, low-resource environments. The present study tests this hypothesis across diverse timeframes, branches of immunity, and conditions of energy availability among humans. Using a prospective mixed-longitudinal design, we collected anthropometric and blood immune biomarker data from 261 Amazonian forager-horticulturalist Shuar children (age 4-11 y old). This strategy provided baseline measures of participant stature, s.c. body fat, and humoral and cell-mediated immune activity as well as subsample longitudinal measures of linear growth (1 wk, 3 mo, 20 mo) and acute inflammation. Multilevel analyses demonstrate consistent negative effects of immune function on growth, with children experiencing up to 49% growth reduction during periods of mildly elevated immune activity. The direct energetic nature of these relationships is indicated by (i) the manifestation of biomarker-specific negative immune effects only when examining growth over timeframes capturing active competition for energetic resources, (ii) the exaggerated impact of particularly costly inflammation on growth, and (iii) the ability of children with greater levels of body fat (i.e., energy reserves) to completely avoid the growth-inhibiting effects of acute inflammation. These findings provide evidence for immunologically and temporally diverse body fat-dependent tradeoffs between immune function and growth during childhood. We discuss the implications of this work for understanding human developmental energetics and the biological mechanisms regulating variation in human ontogeny, life history, and health.

Disparities in bone density across contemporary Amazonian forager-horticulturalists: Cross-population comparison of the Tsimane and Shuar.

OBJECTIVES: This study investigates bone density across the life course among Bolivian Tsimane and Ecuadorian Shuar of Amazonia. Both groups are rural, high-fertility forager-horticulturalists, with high lifetime physical activity levels. We test whether Tsimane and Shuar bone density patterns are different from each other, and if both groups are characterized by lower osteoporosis risk compared to U.S. references. METHODS: Anthropometric and calcaneal bone density data, obtained via quantitative ultrasonometry (QUS), were collected from 678 Tsimane and 235 Shuar (13-92 years old). Population and sex differences in QUS values (estimated bone mineral density, speed of sound, broadband ultrasound attenuation) by age group were assessed using Mann-Whitney U tests. Age-related change and age at peak QUS value were determined using polynomial regressions. One-way analyses of covariance assessed population-level differences in QUS values by age group adjusting for body mass index. Participants aged 50+ years at elevated osteoporosis risk were identified using a T score < -1.8; binomial tests assessed risk compared to U.S. references. RESULTS: Shuar males and females <50 years old have QUS values 3-36% higher than Tsimane, with differences evident in adolescence. Among Tsimane and Shuar, 49 and 23% of participants aged 50+ years old, respectively, are at high risk for osteoporosis, compared to 34% of Americans; Shuar osteoporosis risk is comparable to Americans, while Tsimane risk is elevated. CONCLUSIONS: Disparate patterns in QUS values are documented for Tsimane and Shuar, with pronounced differences early in life. Potential explanations for differences include gene-environment interactions and/or degree of market integration, which influences diet, activity profiles, pathogen exposures, and other lifestyle covariates. As Tsimane osteoporosis risk is greater than in the United States, findings point to alternative risk factors for low bone density that are not readily discernible in industrialized populations.

Soil-transmitted helminth infection and intestinal inflammation among the Shuar of Amazonian Ecuador.

OBJECTIVES: Little research exists documenting levels of intestinal inflammation among indigenous populations where exposure to macroparasites, like soil-transmitted helminths (STHs), is common. Reduced STH exposure is hypothesized to contribute to increased prevalence of elevated intestinal inflammation in wealthy nations, likely due to coevolutionary histories between STHs and human immune systems that favored anti-inflammatory pathways. Here, we document levels of intestinal inflammation and test associations with STH infection among the Shuar of Ecuador, an indigenous population undergoing socioeconomic/lifestyle changes that influence their hygienic environment. We predict that fecal calprotectin (FC; a measure of intestinal inflammation) will be lower in STH infected individuals and that FC will be negatively associated with infection intensity. METHODS: Stool samples to analyze FC levels and STH infection were collected from 69 Shuar participants (ages 5-75 years). Children (<15 years) and adults (15+ years) were analyzed separately to understand the role of exposure in immune system development and the intestinal inflammatory response. RESULTS: Two species of STH were present: Ascaris lumbricoides and Trichuris trichiura. The relationships between infection and intestinal inflammation were age- and species-specific. While no significant relationships were found among adults, children who were singly infected with T. trichiura had lower FC levels than uninfected children. Infection intensity was not significantly associated with FC in children or adults. CONCLUSIONS: These preliminary results provide limited support for our hypotheses, documenting tentative age- and species-specific associations between FC and infection status. Findings may point to the importance of species-specific STH exposure during immune system development.

High energy requirements and water throughput of adult Shuar forager-horticulturalists of Amazonian Ecuador.

OBJECTIVES: We measured total energy expenditure (TEE; kcal/d) and water throughput (L/d) among Shuar forager-horticulturalists from Amazonian Ecuador to compare their daily energy and water demands to adults in other small-scale and industrialized populations. METHODS: TEE and water throughput were measured using the doubly labeled water method among 15 Shuar adults (eight women, seven men; age range 18-60 years) living in a relatively remote village. We used multiple regression to assess the effects of anthropometric variables (body size, fat free mass, age, and sex) on TEE and water throughput. We also compared Shuar TEE and water throughput to those of other small-scale and industrialized societies. RESULTS: TEE among Shuar adults (men: 4141 ± 645 kcal/d, women: 2536 ± 281 kcal/d) was most strongly correlated with fat free mass. Estimated physical activity levels (PAL) calculated as (TEE/estimated BMR), were greater for men (2.34 ± 0.29) than women (1.83 ± 0.14, P < 0.001). Water throughput was also greater among Shuar men (9.37 ± 2.34 L/d) than women (4.76 ± 0.36 L/d, P < 0.001). Shuar TEE and water throughput were elevated compared to adults in industrialized populations. DISCUSSION: TEE and PAL of Shuar men are among the highest recorded during normal daily life, and likely reflect both high levels of physical activity and cultural dietary practices. Drinking large amounts of chicha, a traditional carbohydrate-rich drink made from manioc, likely contributes to the high levels of water throughput among Shuar men, and may contribute to elevated TEE.

Global variation in diurnal cortisol rhythms: evidence from Garisakang forager-horticulturalists of lowland Papua New Guinea.

The hypothalamic-pituitary-adrenal (HPA) axis represents an important and evolutionarily ancient biological pathway linking physical and psychological stressors with human health. Despite considerable research exploring the physiological stress response among developed populations, few studies have examined HPA activity in non-industrialized contexts, restricting understanding of variation in human stress reactivity across global socio-ecological diversity. The present study addresses this shortcoming by investigating diurnal cortisol rhythms among Garisakang forager-horticulturalists of remote, lowland Papua New Guinea. Using a large sample of repeated salivary cortisol measurements from 169 participants (age 4-70 years), multilevel growth curve models were constructed to assess Garisakang waking cortisol concentrations and diurnal cortisol slopes. As predicted, results demonstrate identifiable but substantially diminished diurnal cortisol rhythms relative to those of industrialized populations. Sample-wide, Garisakang cortisol concentrations are highest upon waking (mean = 4.86 nmol/L) and decrease throughout the day at a mean rate of only -0.18 nmol/L/h or -6.20%/h. Age and sex significantly predict evaluated cortisol parameters in ways not consistently reported among industrialized populations, suggesting that Garisakang diurnal cortisol rhythms are defined by distinct ontogenetic trajectories across the lifespan. These findings highlight cross-cultural diversity in HPA activity and have important implications for understanding basic mechanisms of the physiological stress response in contexts of chronic physical stressors such as limited nutrition, heavy burden of infectious disease, and high levels of physical activity.

Growth references for Tsimane forager-horticulturalists of the Bolivian Amazon.

OBJECTIVES: Growth standards and references currently used to assess population and individual health are derived primarily from urban populations, including few individuals from indigenous or subsistence groups. Given environmental and genetic differences, growth may vary in these populations. Thus, there is a need to assess whether international standards are appropriate for all populations, and to produce population specific references if growth differs. Here we present and assess growth references for the Tsimane, an indigenous population of Bolivian forager-horticulturalists. METHODS: Mixed cross-sectional/longitudinal anthropometrics (9,614 individuals; 30,118 observations; ages 0-29 years) were used to generate centile curves and Lambda-Mu-Sigma (LMS) tables for height-for-age, weight-for-age, body mass index (BMI)-for-age, and weight-for-height (WFH) using Generalized Additive Models for Location Shape and Scale (GAMLSS). Velocity curves were generated using SuperImposition by Translation and Rotation (SITAR). Tsimane ≤5 years were compared to World Health Organization (WHO) standards while those >5 years were compared to WHO school age references. All ages were compared to published references for Shuar forager-horticulturalists of the Ecuadorian Amazon. RESULTS: Tsimane growth differs from WHO values in height and weight, but is similar for BMI and WFH. Tsimane growth is characterized by slow height velocity in childhood and early adolescent peak height velocity at 11.3 and 13.2 years for girls and boys. Tsimane growth patterns are similar to Shuar, suggesting shared features of growth among indigenous South Americans. CONCLUSIONS: International references for BMI-for-age and WFH are likely appropriate for Tsimane, but differences in height-for-age and weight-for-age suggest Tsimane specific references may be useful for these measures.

The application of knemometry to measure childhood short-term growth among the indigenous Shuar of Ecuador.

OBJECTIVES: Knemometry, the precise measurement of lower leg (LL) length, suggests that childhood short-term (e.g., weekly) growth is a dynamic, nonlinear process. However, owing to the large size and complexity of the traditional knemometer device, previous study of short-term growth among children has been restricted predominantly to clinical settings in industrialized Western nations. The aim of the present study is to address this limitation and promote broader understandings of global variation in childhood development by: (1) describing a custom-built portable knemometer and assessing its performance in the field; and (2) demonstrating the potential application of such a device by characterizing childhood short-term LL growth among the indigenous Shuar of Amazonian Ecuador. MATERIALS AND METHODS: Mixed-longitudinal LL length data were collected weekly from 336 Shuar children age 5-12 years old using the custom portable knemometer (n = 1,145 total observations). Device performance and Shuar short-term LL growth were explored using linear mixed effects models and descriptive statistics. RESULTS: The portable knemometer performed well across a range of participant characteristics and possesses a low technical error of measurement of 0.18 mm. Shuar childhood LL growth averages 0.47 mm/week (SD = 0.75 mm/week), but exhibits large between- and within-individual variation. DISCUSSION: Knemometry can be reliably performed in the field, providing a means for evaluating childhood short-term growth among genetically and ecologically diverse populations. Preliminary findings suggest that Shuar weekly LL growth is comparable in mean magnitude but likely more variable than reported for healthy Western children. Future work will further explore these patterns. Am J Phys Anthropol 160:353-357, 2016. © 2016 Wiley Periodicals, Inc.

Physical growth of the shuar: Height, Weight, and BMI references for an indigenous amazonian population.

OBJECTIVES: Information concerning physical growth among small-scale populations remains limited, yet such data are critical to local health efforts and to foster basic understandings of human life history and variation in childhood development. Using a large dataset and robust modeling methods, this study aims to describe growth from birth to adulthood among the indigenous Shuar of Amazonian Ecuador. METHODS: Mixed-longitudinal measures of height, weight, and body mass index (BMI) were collected from Shuar participants (n = 2,463; age: 0-29 years). Centile growth curves and tables were created for each anthropometric variable of interest using Generalized Additive Models for Location, Scale, and Shape (GAMLSS). Pseudo-velocity and Lambda-Mu-Sigma curves were generated to further investigate Shuar patterns of growth and to facilitate comparison with United States Center for Disease Control and Prevention and multinational World Health Organization growth references. RESULTS: The Shuar are small throughout life and exhibit complex patterns of growth that differ substantially from those of international references. Similar to other Amazonians, Shuar growth in weight compares more favorably to references than growth in height, resulting in BMI curves that approximate international medians. Several additional characteristics of Shuar development are noteworthy, including large observed variation in body size early in life, significant infant growth faltering, extended male growth into adulthood, and a markedly early female pubertal growth spurt in height. Phenotypic plasticity and genetic selection in response to local environmental factors may explain many of these patterns. CONCLUSIONS: Providing a detailed reference of growth for the Shuar and other Amazonian populations, this study possesses direct clinical application and affords valuable insight into childhood health and the ecology of human growth.

Heterogeneous effects of market integration on sub-adult body size and nutritional status among the Shuar of Amazonian Ecuador.

BACKGROUND: Market integration (MI)-increasing production for and consumption from a market-based economy-is drastically altering traditional ways of life and environmental conditions among indigenous Amazonian peoples. The effects of MI on the biology and health of Amazonian children and adolescents, however, remain unclear. AIM: This study examines the impact of MI on sub-adult body size and nutritional status at the population, regional and household levels among the Shuar of Amazonian Ecuador. SUBJECTS AND METHODS: Anthropometric data were collected between 2005-2014 from 2164 Shuar (aged 2-19 years) living in two geographic regions differing in general degree of MI. High-resolution household economic, lifestyle and dietary data were collected from a sub-sample of 631 participants. Analyses were performed to investigate relationships between body size and year of data collection, region and specific aspects of household MI. RESULTS: Results from temporal and regional analyses suggest that MI has a significant and overall positive impact on Shuar body size and nutritional status. However, household-level results exhibit nuanced and heterogeneous specific effects of MI underlying these overarching relationships. CONCLUSION: This study provides novel insight into the complex socio-ecological pathways linking MI, physical growth and health among the Shuar and other indigenous Amazonian populations.

Effect of chewing betel nut (Areca catechu) on salivary cortisol measurement.

OBJECTIVES: Cultural practices may compromise the accuracy of salivary hormone measurements and must be considered when designing human biology research protocols. This study aims to evaluate the acute effect of one common human practice-chewing betel nut-on the measurement of salivary cortisol levels under field conditions. MATERIALS AND METHODS: Data were collected from 17 adult habitual betel nut users (males = 11; females = 6; mean age = 32.8 years) from a small rural community in Papua New Guinea. Saliva was collected in time series from each participant before and at 0, 15, 30, 45, 60, and 75 min after chewing betel nut. Samples were analyzed by radioimmunoassay and cortisol levels were compared across time using linear mixed effects modeling. RESULTS: Measured mean cortisol concentration fell nearly 40% immediately following betel nut use and remained significantly below baseline levels for the following 45 min (all P < 0.05). Cortisol concentrations measured at 60 min and 75 min were indistinguishable from baseline levels (all P > 0.16). DISCUSSION: Chewing betel nut is associated with a transient but significant reduction in measured levels of salivary cortisol. Future research must take this into account in populations where betel nut use is prevalent.

Refugee health and physiological profiles in transitional settlements in Serbia and Kenya: Comparative evidence for effects of gender and social support.

When armed conflict compels people to flee from their homelands, they embark on protracted journeys during which they experience wide ranging physical, social, and psychological challenges. Few studies have focused on refugee psychosocial and physiological profiles during the transitional phase of forced migration that often involves temporary sheltering. Transient refugees' experiences can vary substantially based on local socio-ecological conditions in temporary settlements, including the length of stay, living conditions, as well as the availability and accessibility of physical and social resources. In this study, we compared physiological and psychosocial data from refugees (N=365; 406 observations) in Serbia and Kenya, respectively, with divergent temporal (length of stay) and socio-ecological conditions. In Serbia, refugees resided in asylum centers (mean stay: 0.9 y); in Kenya they were living in Kakuma Refugee Camp (mean stay: 8.8 y), one of the world's largest camps at the time. We had limited ability to directly compare psychosocial measures and used meta-analytic techniques to evaluate predictors of refugee mental and physical health at the two sites, including based on perceived social support. Refugees in Serbia had higher fingernail cortisol (p < 0.001) and were less likely to have elevated C-reactive protein (CRP) levels (p < 0.01) than refugees in Kakuma. We found common gender differences in both settings; women had lower cortisol but higher EBV antibody titers and higher likelihood of having elevated CRP compared to men (all p < 0.01). Woman also reported poorer mental and physical health (p < 0.001). These physiological and health differences may reflect variation between men and women in their psychosocial and physical experiences of factors such as stress, violence, and trauma during their journeys and as transitional refugees. Finally, we also found that refugees with lower levels of perceived social support reported poorer physical and mental health (p < 0.001). Although our results are cross-sectional, they suggest that this intermittent phase of the refugee experience is a key window for helping enhance refugee well-being through an emphasis on interpersonal and community support systems.