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.

A comparison of faecal glucocorticoid metabolite concentration and gut microbiota diversity in bonobos (Pan paniscus).

Sex, age, diet, stress and social environment have all been shown to influence the gut microbiota. In several mammals, including humans, increased stress is related to decreasing gut microbial diversity and may differentially impact specific taxa. Recent evidence from gorillas shows faecal glucocorticoid metabolite concentration (FGMC) did not significantly explain gut microbial diversity, but it was significantly associated with the abundance of the family Anaerolineaceae. These patterns have yet to be examined in other primates, like bonobos (Pan paniscus). We compared FGMC to 16S rRNA amplicons for 202 bonobo faecal samples collected across 5 months to evaluate the impact of stress, measured with FGMC, on the gut microbiota. Alpha diversity measures (Chao's and Shannon's indexes) were not significantly related to FGMC. FGMC explained 0.80 % of the variation in beta diversity for Jensen-Shannon and 1.2% for weighted UniFrac but was not significant for unweighted UniFrac. We found that genus SHD-231, a member of the family Anaerolinaceae had a significant positive relationship with FGMC. These results suggest that bonobos are relatively similar to gorillas in alpha diversity and family Anaerolinaceae responses to FGMC, but different from gorillas in beta diversity. Members of the family Anaerolinaceae may be differentially affected by FGMC across great apes. FGMC appears to be context dependent and may be species-specific for alpha and beta diversity but this study provides an example of consistent change in two African apes. Thus, the relationship between physiological stress and the gut microbiome may be difficult to predict, even among closely related species.

Minimally invasive biomarkers in human population biology research, part 2: An introduction to the special issue.

Over the past several decades, biomarkers have become indispensable tools in human biology, allowing researchers to investigate biocultural and evolutionary questions and to generate basic data on health and well-being. Human biologists are central players in biomarker methods development, pioneering the creation of "field friendly," minimally invasively collected biomarker approaches, as well as leading the innovative use of biomarkers, most notably to map the complex pathways from social and environmental conditions to altered physiology to health effects. This special issue, Part 2 of a collection focused on minimally invasively collected biomarkers, is comprised of nine papers that jointly contribute to driving the science of biomarker methods development and application. This compilation of papers engages with topics such as biological normalcy, ecoimmunology, and the ethics of working with vulnerable and underserved populations. It also focuses attention on research areas at present not emphasized in human biology such as bone turnover markers as a window onto osteoporosis and osteoarthritis and the use of cancer-related biomarkers in population screening and epidemiology. Taken together, these papers help draw the roadmap for future biomarker work, emphasizing: (1) the need for systematic and transparent approaches to assay validation, with open access publication; (2) simultaneous measurement of multiple biomarkers and expanded use of instrument platforms that increase the range of physiological, genomics, and omics markers available to researchers; and, (3) increased attention to ethics and researcher responsibilities, encouraging a mindset that recognizes our obligation to provide benefits to individuals and communities and to help redress past abuses.

The utility of dried blood spot measurement of bone turnover markers in biological anthropology.

OBJECTIVES: Bone is a dynamic organ under continual turnover influenced by life history stage, energy dynamics, diet, climate, and disease. Bone turnover data have enormous potential in biological anthropology for testing evolutionary and biocultural hypotheses, yet few studies have integrated these biomarkers. In the present article we systematically review the current availability, future viability, and applicability of measuring bone turnover markers (BTMs) in dried blood spot (DBS) samples obtained from finger prick whole blood. METHODS: Our review considers clinical and public health relevance, biomarker stability in DBS, assay availability, and cost. We consider biomarkers of bone formation such as osteocalcin (bone matrix protein), PINP (N-terminal propeptide of type I collagen), and alkaline phosphatase (osteoblast enzyme), as well as biomarkers of bone resorption such as CTX (marker of collagen breakdown) and TRACP5b (tartrate-resistant acid phosphatase 5b; osteoclast enzyme). RESULTS: Two BTMs have been validated for DBS: osteocalcin (formation) and TRACP5b (resorption). Prime candidates for future development are CTX and PINP, the formation and resorption markers used for clinical monitoring of response to osteoporosis treatment. CONCLUSION: BTMs are a field-friendly technique for longitudinal monitoring of skeletal biology during growth, reproduction and aging, combining minimized risk to study participants with maximized ease of sample storage and transport. This combination allows new insights into the effects of energy availability, disease, and physical activity level on bone, and questions about bone gain and loss across life history and in response to environmental factors; these issues are important in human biology, paleoanthropology, bioarchaeology, and forensic anthropology.

Current and future applications of biomarkers in samples collected through minimally invasive methods for cancer medicine and population-based research.

Despite advances in cancer medicine and research, invasive and potentially risky procedures such as biopsies, venous blood tests, imaging, colonoscopy, and pap smear tests are still primarily used for screening, staging, and assessing response to therapy. The development and interdisciplinary use of biomarkers from urine, feces, saliva, scent, and capillary blood collected with minimally invasive methods represents a potential opportunity for integration with biomarker analysis for cancers, both in clinical practice (e.g., in screening, treatment, and disease monitoring, and improved quality of life for patients) and population-based research (e.g., in epidemiology/public health, studies of social and environmental determinants, and evolutionary medicine). In this article, we review the scientific rationale, benefits, challenges, and potential opportunities for measuring cancer-related biomarkers in samples collected through minimally invasive methods.

After Theranos: Using point-of-care testing to advance measures of health biomarkers in human biology research.

OBJECTIVES: The rise and fall of the health technology startup Theranos is emblematic of the promise and peril of point-of-care testing (POCT). Instruments that deliver immediate results from minimally invasive samples at the location of collection can provide powerful tools to deliver health data in clinical and public health contexts. Yet, POCT availability is driven largely by market interests, which limits the development of inexpensive tests for diverse health conditions that can be used in resource-limited settings. These constraints, combined with complex regulatory hurdles and substantial ethical challenges, have contributed to the underutilization of POCT in human biology research. METHODS: We evaluate current POCT capabilities and limitations, discuss promising applications for POCT devices in resource-limited settings, and discuss the future of POCT. RESULTS: As evidenced by publication trends, POCT platforms have rapidly advanced in recent years, gaining traction among clinicians and health researchers. We highlight POCT devices of potential interest to population-based researchers and present specific examples of POCT applications in human biology research. CONCLUSIONS: Several barriers can limit POCT applications, including cost, lack of regulatory approval for non-clinical use, requirements for expensive equipment, and the dearth of validation in remote field conditions. Despite these issues, we see immense potential for emerging POCT technology capable of analyzing new sample types and used in conjunction with increasingly common technology (e.g., smart phones). We argue that the fallout from Theranos may ultimately provide an opportunity to advance POCT, leading to more ethical data collection and novel opportunities in human biology research.

Transient refugees' social support, mental health, and physiological markers: Evidence from Serbian asylum centers.

OBJECTIVES: Refugees seeking safety across international borders are often exposed to a wide breadth of psychosocially stressful experiences that may fracture existing sources of social support and impair the generation of new social relationships, with implications for their long-term health and resilience. Using data from recently settled refugees in two asylum centers in Serbia, we examined the associations between social support, mental health, and physiological markers. METHODS: In this mixed-method study of refugees (age 18-50 years, n = 76), we collected key socio-demographic information and conducted semi-structured interviews about refugees' journey and stay in Serbia, trauma/loss, and their sources of social support. We also collected self-reported measures of mental well-being as well as physiological markers relevant to repeated exposure to chronic psychosocial stress (fingernail cortisol and dried blood spots for analysis of Epstein-Barr virus [EBV] antibody titers). RESULTS: We found that refugees with longer journeys reported lower social support than those with shorter journeys. Refugees with lower social support reported poorer mental well-being, greater PTSD-related symptoms, and higher recent perceived stress than those with higher social support. We also observed that refugees with lower social support and higher recent stress, respectively, tended to exhibit higher fingernail cortisol levels. However, we did not observe comparable patterns linking EBV antibodies with psychosocial functioning. CONCLUSION: Our cross-sectional findings are consistent with the notion that social support is likely to be a critical component in effective interventions aimed at mitigating the adverse health effects of relocation-related illnesses and poor social functioning as they await resettlement.

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.

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.

Cross-cultural invariances in the architecture of shame.

Human foragers are obligately group-living, and their high dependence on mutual aid is believed to have characterized our species' social evolution. It was therefore a central adaptive problem for our ancestors to avoid damaging the willingness of other group members to render them assistance. Cognitively, this requires a predictive map of the degree to which others would devalue the individual based on each of various possible acts. With such a map, an individual can avoid socially costly behaviors by anticipating how much audience devaluation a potential action (e.g., stealing) would cause and weigh this against the action's direct payoff (e.g., acquiring). The shame system manifests all of the functional properties required to solve this adaptive problem, with the aversive intensity of shame encoding the social cost. Previous data from three Western(ized) societies indicated that the shame evoked when the individual anticipates committing various acts closely tracks the magnitude of devaluation expressed by audiences in response to those acts. Here we report data supporting the broader claim that shame is a basic part of human biology. We conducted an experiment among 899 participants in 15 small-scale communities scattered around the world. Despite widely varying languages, cultures, and subsistence modes, shame in each community closely tracked the devaluation of local audiences (mean r = +0.84). The fact that the same pattern is encountered in such mutually remote communities suggests that shame's match to audience devaluation is a design feature crafted by selection and not a product of cultural contact or convergent cultural evolution.

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.

Validation of an enzyme-linked immunoassay assay for osteocalcin, a marker of bone formation, in dried blood spots.

OBJECTIVES: Investigating factors that contribute to bone loss and accretion across populations in remote settings is challenging, particularly where diagnostic tools are scarce. To mitigate this challenge, we describe validation of a commercial ELISA assay to measure osteocalcin, a biomarker of bone formation, from dried blood spots (DBS). METHODS: We validated the Osteocalcin Human SimpleStep ELISA kit from Abcam (ab1951214) using 158 matched plasma and DBS samples. Passing-Bablok regression analysis assessed the relationships between plasma and DBS osteocalcin concentrations. Dilutional linearity and spike and recovery experiments determined if the DBS matrix interfered with osteocalcin measurement, and intra- and inter-assay coefficients of variation (CVs) were calculated. Limit of detection, analyte stability, and specific forms of osteocalcin measured by the kit were also investigated. RESULTS: Mean plasma osteocalcin value was 218.2 ng/mL (range 64.6-618.1 ng/mL). Linear relationships existed between plasma and DBS concentrations of osteocalcin, with no apparent bias in plasma vs DBS concentrations. There was no apparent interference of the DBS matrix with measurement of osteocalcin in DBS. Intra-assay CV for DBS was ~8%, while average inter-assay CV was 14.8%. Limit of detection was 0.34 ng/mL. Osteocalcin concentrations were stable in DBS stored at -28°C and room temperature, but not those stored at 37°C. This ELISA kit detects total osteocalcin. CONCLUSIONS: Osteocalcin, a bone formation biomarker, can be measured from DBS. Combined with a previously validated DBS assay for TRACP-5b, a bone resorption biomarker, these assays have the potential to help researchers disentangle the many factors contributing to bone strength.

Inflammation and central adiposity as mediators of depression and uncontrolled diabetes in the study on global AGEing and adult health (SAGE).

OBJECTIVES: Diabetes and depression are commonly present in the same individuals, suggesting the possibility of underlying shared physiological processes. Inflammation, as assessed with the biomarker C-reactive protein (CRP), has not consistently explained the observed relationship between diabetes and depression, although both are associated with inflammation and share proposed inflammatory mechanisms. Central adiposity has also been associated with both conditions, potentially by causing increased inflammation. This study uses the World Health Organization's Study on global AGEing and adult health (SAGE) Mexico Wave 1 biomarker data (n = 1831) to evaluate if inflammation and central adiposity mediate the relationship between depression and diabetes. METHODS: Depression was estimated using a behavior-based diagnostic algorithm, inflammation using venous dried blood spot (DBS) CRP, central adiposity using waist-to-height ratio (WHtR), and uncontrolled diabetes using venous DBS-glycated hemoglobin (HbA1c). RESULTS: The association between depression and uncontrolled diabetes was partially mediated by CRP before but not after WHtR was considered. When WHtR was added to the model, it partially mediated the relationship between diabetes and depression while fully mediating the relationship between depression and CRP. CONCLUSIONS: These findings suggest that central adiposity may be a more significant mediator between diabetes and depression than inflammation and account for the relationship between these disorders and inflammation. Depression may cause an increase in central adiposity, which then may lead to diabetes, but the increase in known systemic inflammatory pathways caused by central adiposity may not be the key pathological mechanism.

Epigenetic aging in children from a small-scale farming society in The Congo Basin: Associations with child growth and family conflict.

Developmental environments influence individuals' long-term health trajectories, and there is increasing emphasis on understanding the biological pathways through which this occurs. Epigenetic aging evaluates DNA methylation at a suite of distinct CpG sites in the genome, and epigenetic age acceleration (EAA) is linked to heightened chronic morbidity and mortality risks in adults. Consequently, EAA provides insights on trajectories of biological aging, which early life experiences may help shape. However, few studies have measured correlates of children's epigenetic aging, especially outside of the U.S. and Europe. In particular, little is known about how children's growth and development relate to EAA in ecologies in which energetic and pathogenic stressors are commonplace. We studied EAA from dried blood spots among Bondongo children (n = 54) residing in a small-scale, fisher-farmer society in a remote region of the Republic of the Congo. Here, infectious disease burdens and their resultant energy demands are high. Children who were heavier for height or taller for age, respectively, exhibited greater EAA, including intrinsic EAA, which is considered to measure EAA internal to cells. Furthermore, we found that children in families with more conflict between parents had greater intrinsic EAA. These results suggest that in contexts in which limited energy must be allocated to competing demands, more investment in growth may coincide with greater EAA, which parallels findings in European children who do not face similar energetic constraints. Our findings also indicate that associations between adverse family environments and greater intrinsic EAA were nonetheless observable but only after adjustment for covariates relevant to the energetically and immunologically demanding nature of the local ecology.

Knee osteoarthritis risk in non-industrial societies undergoing an energy balance transition: evidence from the indigenous Tarahumara of Mexico.

Non-industrial societies with low energy balance levels are expected to be less vulnerable than industrial societies to diseases associated with obesity including knee osteoarthritis. However, as non-industrial societies undergo rapid lifestyle changes that promote positive energy balance, individuals whose metabolisms are adapted to energetic scarcity are encountering greater energy abundance, increasing their propensity to accumulate abdominal adipose tissue and thus potentially their sensitivity to obesity-related diseases. OBJECTIVES: Here, we propose that knee osteoarthritis is one such disease for which susceptibility is amplified by this energy balance transition. METHODS: Support for our hypothesis comes from comparisons of knee radiographs, knee pain and anthropometry among men aged ≥40 years in two populations: Tarahumara subsistence farmers in Mexico undergoing the energy balance transition and urban Americans from Framingham, Massachusetts. RESULTS: We show that despite having markedly lower obesity levels than the Americans, the Tarahumara appear predisposed to accrue greater abdominal adiposity (ie, larger abdomens) for a given body weight, and are more vulnerable to radiographic and symptomatic knee osteoarthritis at lower levels of body mass index. Also, proportionate increases in abdomen size in the two groups are associated with greater increases in radiographic knee osteoarthritis risk among the Tarahumara than the Americans, implying that the abdominal adipose tissue of the Tarahumara is a more potent stimulus for knee degeneration. CONCLUSIONS: Heightened vulnerability to knee osteoarthritis among non-industrial societies experiencing rapid lifestyle changes is a concern that warrants further investigation since such groups represent a large but understudied fraction of the global population.

A dried blood spot-based method to measure levels of tartrate-resistant acid phosphatase 5b (TRACP-5b), a marker of bone resorption.

OBJECTIVES: A number of basic questions about bone biology have not been answered, including population differences in bone turnover. In part, this stems from the lack of validated minimally invasive biomarker techniques to measure bone formation and resorption in field-based population-level research. The present study addresses this gap by validating a fingerprick dried blood spot (fDBS) assay for tartrate-resistant acid phosphatase 5b (TRACP-5b), a well-defined biomarker of bone resorption and osteoclast number. METHODS: We adapted a commercially available enzyme-linked immunosorbent assay (ELISA) kit from MyBiosource for the quantitative determination of TRACP-5b levels in serum and plasma for use with DBS. We used a rigorous process of assay modification and validation, including the use of a matched set of 189 adult plasma, fDBS, and venous DBS (vDBS) samples; parameters evaluated included precision, reliability, and analyte stability. RESULTS: Plasma and DBS TRACP-5b concentrations showed a linear relationship. There were no systematic differences in TRACP-5b levels in fDBS and vDBS, indicating no significant differences in TRACP-5b distribution between capillary and venous blood. Parallelism and spike-and-recovery results indicated that matrix factors in DBS do not interfere with measurement of TRACP-5b levels from DBS using the validated kit. Intra- and interassay CVs were 5.0% and 12.1%, respectively. DBS samples should preferably be stored frozen but controlled room temperature storage for up to a month may be acceptable. CONCLUSIONS: This DBS-based ELISA assay adds to the methodological toolkit available to human biologists and will facilitate research on bone turnover in population studies.

Climbing the ladder of decline: Income and acculturation associated with chronic inflammation among Mexican immigrants.

OBJECTIVES: Financial hardship and immigrant status are often associated with poorer health as immigrant groups acculturate to life in the US. Known as the Latino health paradox, studies have shown that Latino/a immigrants in particular often experience declines in health the more they embrace ways of life considered "dominant" by US society. At present, critical biological pathways linking socioeconomic and acculturative processes remain to be better explained. The present study investigates associations among financial strain, acculturation, and chronic inflammation. METHODS: In our study of 129 Mexican-born immigrants living in the US, we used Pearson's correlations and multiple regression analyses to investigate links among income-to-poverty ratio (an indicator of financial strain), English language engagement (acculturation), and C-reactive protein (CRP), a measure of systemic inflammation. RESULTS: Results showed that for men, but not women, acculturation as defined by English language engagement moderated the association between an income-to-poverty ratio and CRP levels. CONCLUSIONS: Consistent with the Latino health paradox, more acculturated men with relatively higher income levels (compared with the study sample) had significantly higher levels of CRP.

Economic comparison of systemic antimicrobial therapies for metritis in dairy cows.

Metritis is a prevalent disease with effects on production, reproduction, and survival, thereby affecting dairy farm profitability. A component of the cost of metritis is antimicrobial therapy. Some antimicrobials result in milk withhold that adds to the cost of disease. The objectives were (1) to determine cost of metritis for 2 antimicrobial treatments using a herd budget that includes costs associated with incidence of concurrent diseases, milk production and reproduction losses, and removal from the herd and (2) to apply sensitivity analysis to determine the cost of different scenarios. Cows with metritis from a previous study assigned randomly to receive ampicillin (AMP, n = 259), an antimicrobial that requires milk withhold, or ceftiofur (CEFT, n = 269), an antimicrobial with no milk withhold, were used for the economic analysis. A group of cows with no metritis (NOMET, n = 268), matched by parity and calving day, served as the baseline for comparison. The incidence of other diseases in the first 60 d postpartum, culling and death, reproductive performance, discarded milk, milk yield, total milk sold per cow, and residual cow value were used as responses. The economic analysis considered the costs associated with therapy, reproductive management, discarded milk, estimated DM consumed, income from saleable milk, and the residual cow value at 300 d postpartum or earlier if the cow was removed from the herd. Sensitivity analyses were performed considering 3 scenarios for milk and feed prices. The incidence of diseases other than metritis (NOMET, 30.4%; AMP, 45.4%; CEFT, 34.0%) and days in the hospital (NOMET, 2.7 d; AMP, 8.6 d; CEFT, 3.5 d) were greater for cows treated with AMP than CEFT. Treatment did not affect the risk of leaving the herd (NOMET, 15.5%; AMP, 15.0%; CEFT, 19.1%). The 21-d pregnancy rate was lower for cows with metritis but did not differ between AMP and CEFT (NOMET, 24.9%; AMP, 18.9%; CEFT, 17.0%). Milk yield was greater for cows without metritis than those with metritis and greater for AMP than CEFT (NOMET, 33.7 kg/d; AMP, 32.5 kg/d; CEFT, 31.2 kg/d). Cost of metritis did not differ with choice of therapy, but it increased as milk price increased. When both milk and feed prices were the average values considered ($0.44/kg and $0.26/kg, respectively), the costs of a case of metritis for AMP and CEFT were, respectively, $344 and $410 when milk was discarded and $267 and $406 when milk was fed to calves. Cost of therapy for AMP and CEFT represented 16.6 and 24.6% of the total cost of metritis when milk was discarded and 21.5 and 24.8% of the total cost of metritis when milk was fed to calves. The largest component of cost of metritis for both therapies was the reduced income from milk minus feed cost, ranging from 40.0 to 56.7%. Collectively, metritis is an expensive disease, and choice of antimicrobial therapy did not influence survival, reproduction, or cost of the disease.