Observer agreement on the morphology of porous cranial lesions: Results from a workshop at the 2019 meeting of the Paleopathology Association.

OBJECTIVE: This paper presents the results of a 2019 Paleopathology Association workshop that tested observer agreement on porous cranial lesion morphology and presence using multiple sets of existing guidelines for data collection. MATERIALS: Sixteen conference attendees of varying osteological experience served as observers. Three crania were assigned to each of four published guidelines for identifying and categorizing lesion morphology, for a total of twelve well-preserved human crania from the National Museum of Natural History Biological Anthropology Collections. METHODS: Observers assessed each cranium macroscopically according to its assigned set of guidelines. RESULTS: Observer concordance was higher using scoring guidelines with higher-quality photographs, such as the 2019 guidelines from Rinaldo and colleagues. CONCLUSIONS: Data collection guidelines with high-quality color photos may support greater reliability of researcher-generated data on macroscopic skeletal features. SIGNIFICANCE: The conclusions of any research study are only as reliable as the data on which they are based. This work highlights the need for ongoing practices of quality control in a field in which much data results from individual judgement calls. LIMITATIONS: Observer concordance is not a measure of observer accuracy. Sample size is insufficient to draw broadly generalizable conclusions on the reliability of data collected using the guidelines tested, and conference environments are not a facsimile of research settings. SUGGESTIONS FOR FURTHER RESEARCH: Iterative testing of methodological consistency using larger sample sizes and more non-pathological crania is advised to identify the factors that influence observer discordance and to improve guidelines for qualitative assessments.

Skeletal manifestations of disease experience: Length of illness and porous cranial lesion formation in a contemporary juvenile mortality sample.

OBJECTIVES: Porous lesions of the orbit (cribra orbitalia [CO]) and cranial vault (porotic hyperostosis [PH]) are used as skeletal indicators of childhood stress. Because they are understudied in contemporary populations, their relationship to disease experience is poorly understood. This paper examines the relationship between length of childhood illness and CO/PH formation in a clinically documented sample. "Turning points," which identify the window for lesion formation for CO/PH, are defined, implications for hidden heterogeneity in frailty are considered. METHODS: Data are from 333 (199 males; 134 females) pediatric postmortem computed tomography scans. Individuals died in New Mexico (2011-2019) and are 0.5 to 15.99 years (mean = 7.1). Length of illness was estimated using information from autopsy and field reports. Logistic regression was used to estimate predicted probabilities, odds ratios, and the temporal window for lesion formation. RESULTS: Illness, single bouts, or cumulative episodes lasting over 1 month is associated with higher odds of CO; individuals who were never sick have lower odds of having PH. This relationship was consistent for fatal and incidental illnesses that did not cause death. The developmental window for CO formation appears to close at 8 years. CONCLUSIONS: Those ill for over 1 month are more likely to have CO/PH than those with acute illnesses. Some individuals lived sufficiently long to form CO/PH but died of illness. Others with lesions died of circumstances unrelated to disease. This indicates hidden variation in robusticity even among ill individuals with CO/PH, which is vital in interpreting lesion frequencies in the archeological record.

A biological approach to adult sex differences in skeletal indicators of childhood stress.

OBJECTIVES: In previous work examining the etiology of cribra orbitalia (CO) and porotic hyperostosis (PH) in a contemporary juvenile mortality sample, we noted that males had higher odds of having CO lesions than females. Here, we examine potential reasons for this pattern in greater detail. Four non-mutually exclusive mechanisms could explain the observed sex differences: (1) sex-biased mortality; (2) sexual dimorphism in immune responses; (3) sexual dimorphism in bone turnover; or (4) sexual dimorphism in marrow conversion. SUBJECTS AND METHODS: The sample consists of postmortem computed tomography scans and autopsy reports, field reports, and limited medical records of 488 individuals from New Mexico (203 females; 285 males) aged between 0.5 and 15 years. We used Kaplan-Meier survival analysis, predicted probabilities, and odds ratios to test each mechanism. RESULTS: Males do not have lower survival probabilities than females, and we find no indications of sex differences in immune response. Overall, males have a higher probability of having CO or PH lesions than females. CONCLUSIONS: All results indicate that lesion formation in juveniles is influenced by some combination of sex differences in the pace of red-yellow conversion of the bone marrow and bone turnover. The preponderance of males with CO and PH likely speaks to the potential for heightened osteoblastic activity in males. We find no support for the hypotheses that sex biases in mortality or immune responses impacted lesion frequency in this sample. Sex differences in biological processes experienced by children may affect lesion formation and lesion expression in later life.

Do computed tomography findings agree with traditional osteological examination? The case of porous cranial lesions.

OBJECTIVE: The current study evaluates the feasibility of using clinical cranial computed tomography (CT) scans for assessing the presence and morphology of porous cranial lesions (cribra orbitalia, porotic hyperostosis). METHODS: Observers (n = 4) conducted three independent evaluations of porous cranial lesions based on photographs, 2-D CT, and 3-D CT scans of archaeological crania. Evaluations of the crania from each viewing scenario were compared to findings from direct macroscopic observation. MATERIALS: Twenty-two complete adult crania from the Peruvian sites of Pachacamac and Chicama. RESULTS: We found that lesion visibility differed by location: vault lesions with porosity larger than the resolution of the CT scan were identifiable across all viewing scenarios, but orbital lesions were identifiable only when extensive porosity was accompanied by widening of the inter-trabecular spaces. Lesions in stages of advanced remodeling were not visible on CT. CONCLUSIONS: Paleopathological criteria applied to head CTs from clinical cases of suspected cranial fracture can reliably identify moderate to severe porous cranial lesions in living individuals. SIGNIFICANCE: This validation study opens the door to broader study of porous cranial lesions in living individuals that can address open questions about the causes and consequences of these commonly reported skeletal indicators of stress. LIMITATIONS: Performance of all viewing scenarios was evaluated relative to assessment data from direct observation of skeletal remains, but direct observation is itself subject to error. SUGGESTIONS FOR FURTHER RESEARCH: The increasing resolution of routine CTs makes it increasingly possible to explore skeletal lesions in clinical contexts.

Using a multimethod life history approach to navigate the osteological paradox: A case study from Prehispanic Nasca, Peru.

OBJECTIVES: We leverage recent bioarchaeological approaches and life history theory to address the implications of the osteological paradox in a study population. The goal of this article is to evaluate morbidity and mortality patterns as well as variability in the risk of disease and death during the Late Intermediate period (LIP; 950-1450 C.E.) in the Nasca highlands of Peru. We demonstrate how the concurrent use of multiple analytical techniques and life history theory can engage the osteological paradox and provide salient insights into the study of stress, frailty, and resilience in past populations. MATERIALS AND METHODS: Crania from LIP burial contexts in the Nasca highlands were examined for cribra orbitalia (n = 325) and porotic hyperostosis (n = 270). All age groups and both sexes are represented in the sample. Survivor/nonsurvivor analysis assessed demographic differences in lesion frequency and severity. Hazard models were generated to assess differences in survivorship. The relationship between dietary diversity and heterogeneity in morbidity was assessed using stable δ15 N and δ13 C isotope values for bone collagen and carbonate. One hundred and twenty-four crania were directly AMS radiocarbon dated, allowing for a diachronic analysis of morbidity and mortality. RESULTS: The frequency and expression of both orbital and vault lesions increases significantly during the LIP. Survivor/nonsurvivor analysis indicates cranial lesions co-vary with frailty rather than robusticity or longevity. Hazard models show (1) decreasing survivorship with the transition into the LIP, (2) significantly lower adult life expectancy for females compared to males, and (3) individuals with cranial lesions have lower survivorship across the life course. Stable isotope results show very little dietary diversity. Mortality risk and frequency of pathological skeletal lesions were highest during Phase III (1300-1450 C.E.) of the LIP. CONCLUSION: Results provide compelling evidence of increasing physiological stress and mortality in the Nasca highlands during the LIP, but also reveal substantial heterogeneity in frailty and the risk of death. Certain members of society experienced a heavier disease burden and higher mortality compared to their contemporaries. Elevated levels of disease and lethal trauma among females account for some of the sex differences in survivorship but cannot explain the large degree of female-biased mortality. We hypothesize that parental investment in males or increased female fertility rates may explain these differences.

Immune function during pregnancy varies between ecologically distinct populations.

BACKGROUND AND OBJECTIVES: Among placental mammals, females undergo immunological shifts during pregnancy to accommodate the fetus (i.e. fetal tolerance). Fetal tolerance has primarily been characterized within post-industrial populations experiencing evolutionarily novel conditions (e.g. reduced pathogen exposure), which may shape maternal response to fetal antigens. This study investigates how ecological conditions affect maternal immune status during pregnancy by comparing the direction and magnitude of immunological changes associated with each trimester among the Tsimane (a subsistence population subjected to high pathogen load) and women in the USA. METHODOLOGY: Data from the Tsimane Health and Life History Project (N = 935) and the National Health and Nutrition Examination Survey (N = 1395) were used to estimate population-specific effects of trimester on differential leukocyte count and C-reactive protein (CRP), a marker of systemic inflammation. RESULTS: In both populations, pregnancy was associated with increased neutrophil prevalence, reduced lymphocyte and eosinophil count and elevated CRP. Compared to their US counterparts, pregnant Tsimane women exhibited elevated lymphocyte and eosinophil counts, fewer neutrophils and monocytes and lower CRP. Total leukocyte count remained high and unchanged among pregnant Tsimane women while pregnant US women exhibited substantially elevated counts, resulting in overlapping leukocyte prevalence among all third-trimester individuals. CONCLUSIONS AND IMPLICATIONS: Our findings indicate that ecological conditions shape non-pregnant immune baselines and the magnitude of immunological shifts during pregnancy via developmental constraints and current trade-offs. Future research should investigate how such flexibility impacts maternal health and disease susceptibility, particularly the degree to which chronic pathogen exposure might dampen inflammatory response to fetal antigens. LAY SUMMARY: This study compares immunological changes associated with pregnancy between the Tsimane (an Amazonian subsistence population) and individuals in the USA. Results suggest that while pregnancy enhances non-specific defenses and dampens both antigen-specific immunity and parasite/allergy response, ecological conditions strongly influence immune baselines and the magnitude of shifts during gestation.

Cribra orbitalia and porotic hyperostosis are associated with respiratory infections in a contemporary mortality sample from New Mexico.

OBJECTIVES: Cribra orbitalia (CO) and porotic hyperostosis (PH) are porous cranial lesions (PCLs) classically associated with iron-deficiency anemia in bioarchaeological contexts. However, recent studies indicate a need to reassess the interpretation of PCLs. This study addresses the potential health correlates of PCLs in a contemporary sample by examining relationships between the known cause of death (COD) and PCL presence/absence. METHODS: This study includes a sample of 461 juvenile individuals (6 months to 15 years of age) who underwent examination at the University of New Mexico's Office of the Medical Investigator between 2011 and 2019. The information available for each individual includes their sex, age at death, and their COD and manner of death. RESULTS: Odds ratio of having CO (OR = 3.92, p < .01) or PH (OR = 2.86, p = .02) lesions are increased in individuals with respiratory infections. Individuals with heart conditions have increased odds of having CO (OR = 3.52, p = .03) lesions, but not PH. CONCLUSION: Individuals with respiratory infection are more likely to have CO and/or PH. CO appears to have a greater range of health correlates than PH does, as indicated by the heart condition results. However, individuals with congenital heart defects are at higher risk for respiratory infections, so bony alterations in cases of heart conditions may be due to respiratory illness. Since respiratory infection remains a leading cause of mortality today, CO and PH in bioarchaeological contexts should be considered as potential indicators of respiratory infections in the past.

Old friends and friendly fire: Pregnancy, hookworm infection, and anemia among tropical horticulturalists.

OBJECTIVES: Despite public health concerns about hookworm infection in pregnancy, little is known about immune profiles associated with hookworm (Necator americanus and Ancylostoma duodenale) infection during pregnancy. Fetal tolerance requirements may constrain maternal immune response to hookworm, thereby increasing susceptibility to new infections or increasing hemoglobin loss. To explore this possibility, we study systemic immune response and hemoglobin levels in a natural fertility population with endemic helminthic infection. METHODS: We used Bayesian multilevel models to analyze mixed longitudinal data on hemoglobin, hookworm infection, reproductive state, eosinophils, and erythrocyte sedimentation rate (ESR) to examine the effects of pregnancy and hookworm infection on nonspecific inflammation, cellular parasite response, and hemoglobin among 612 Tsimane women aged 15-45 (1016 observations). RESULTS: Pregnancy is associated with lower eosinophil counts and lower eosinophil response to hookworm, particularly during the second and third trimesters. Both hookworm and pregnancy are associated with higher ESR, with evidence for an interaction between the two causing further increases in the first trimester. Pregnancy is moderately associated with higher odds of hookworm infection (OR: 1.23, 95% CI: 0.83 to 1.83). Pregnancy and hookworm both decrease hemoglobin and may interact to accentuate this effect in the first-trimester of pregnancy (Interaction: ß: -0.30 g/dL; CI: -0.870 to 0.24). CONCLUSIONS: Our findings are consistent with a possible trade-off between hookworm immunity and successful pregnancy, and with the suggestion that hookworm and pregnancy may have synergistic effects, particularly in the first trimester.

A microRNA of infectious laryngotracheitis virus can downregulate and direct cleavage of ICP4 mRNA.

Viral microRNAs regulate gene expression using either translational repression or mRNA cleavage and decay. Two microRNAs from infectious laryngotracheitis virus (ILTV), iltv-miR-I5 and iltv-miR-I6, map antisense to the ICP4 gene. Post-transcriptional repression by these microRNAs was tested against a portion of the ICP4 coding sequence cloned downstream of firefly luciferase. Luciferase activity was downregulated by approximately 60% with the iltv-miR-I5 mimic. Addition of an iltv-miR-I5 antagomiR or mutagenesis of the target seed sequence alleviated this effect. The iltv-miR-I5 mimic, when co-transfected with a plasmid expressing ICP4, reduced ICP4 transcript levels by approximately 50%, and inhibition was relieved by an iltv-miR-I5 antagomiR. In infected cells, iltv-miR-I5 mediated cleavage at the canonical site, as indicated by modified RACE analysis. Thus, in this system, iltv-miR-I5 decreased ILTV ICP4 mRNA levels via transcript cleavage and degradation. Downregulation of ICP4 could impact the balance between the lytic and latent states of the virus in vivo.

MicroRNAs of Gallid and Meleagrid herpesviruses show generally conserved genomic locations and are virus-specific.

Many herpesviruses, including Marek's disease viruses (MDV1 and MDV2), encode microRNAs. In this study, we report microRNAs of two related herpesviruses, infectious laryngotracheitis virus (ILTV) and herpesvirus of turkeys (HVT), as well as additional MDV2 microRNAs. The genome locations, but not microRNA sequences, are conserved among all four of these avian herpesviruses. Most are clustered in the repeats flanking the unique long region (I/TR(L)), except in ILTV which lacks these repeats. Two abundant ILTV microRNAs are antisense to the immediate early gene ICP4. A homologue of host microRNA, gga-miR-221, was found among the HVT microRNAs. Additionally, a cluster of HVT microRNAs was found in a region containing two locally duplicated segments, resulting in paralogous HVT microRNAs with 96-100% identity. The prevalence of microRNAs in the genomic repeat regions as well as in local repeats suggests the importance of genetic plasticity in herpesviruses for microRNA evolution and preservation of function.

Computed tomography coronary angiographic demonstration of a single coronary artery with the right coronary artery arising as a continuation from the left circumflex coronary artery.

We present the case of a 31-year-old man undergoing a chest pain evaluation. This report illustrates how 64-slice computed tomographic coronary angiography (CTCA) can aid in the management of patients with equivocal stress test results. It also describes how CTCA can accurately determine the origin and course of an anomalous coronary artery.

UNC-83 IS a KASH protein required for nuclear migration and is recruited to the outer nuclear membrane by a physical interaction with the SUN protein UNC-84.

UNC-84 is required to localize UNC-83 to the nuclear envelope where it functions during nuclear migration. A KASH domain in UNC-83 was identified. KASH domains are conserved in the nuclear envelope proteins Syne/nesprins, Klarsicht, MSP-300, and ANC-1. Caenorhabditis elegans UNC-83 was shown to localize to the outer nuclear membrane and UNC-84 to the inner nuclear membrane in transfected mammalian cells, suggesting the KASH and SUN protein targeting mechanisms are conserved. Deletion of the KASH domain of UNC-83 blocked nuclear migration and localization to the C. elegans nuclear envelope. Some point mutations in the UNC-83 KASH domain disrupted nuclear migration, even if they localized normally. At least two separable portions of the C-terminal half of UNC-84 were found to interact with the UNC-83 KASH domain in a membrane-bound, split-ubiquitin yeast two-hybrid system. However, the SUN domain was essential for UNC-84 function and UNC-83 localization in vivo. These data support the model that KASH and SUN proteins bridge the nuclear envelope, connecting the nuclear lamina to cytoskeletal components. This mechanism seems conserved across eukaryotes and is the first proposed mechanism to target proteins specifically to the outer nuclear membrane.

Effects of body weight gain reduction resulting from chemopreventive agent treatment on mammary gland morphology.

Moderate reductions (< or = 15%) in body weight gain, similar to those observed after administration of some chemopreventive agents in chemically induced mammary cancer models, will result in decreased mammary cancers (up to 55%). The objective of this study was to determine whether changes in mammary gland differentiation, proliferation, apoptosis, and estradiol and progesterone levels are affected by moderate reductions in body weight induced after chemopreventive agent treatment and dietary restriction. The body weights of female Sprague-Dawley rats were reduced by dietary restrictions to match those of rats receiving 4-hydroxyphenylretinamide (4-HPR) at a dose known to inhibit methylnitrosourea (MNU)-induced mammary cancers. 4-HPR supplementation or dietary restrictions began 1 wk before MNU administration at 50 days of age. Mammary gland differentiation, proliferation, apoptosis, and serum levels of estradiol and progesterone were measured at 50, 57, and 71 days of age. Casein expression, proliferating cellular nuclear antigen expression, and apoptosis were not significantly different from controls in the dietary-restricted group. Proliferating cellular nuclear antigen expression was significantly lower in 4-HPR-treated animals than in controls at 57 days of age. The diameter of the mammary gland ducts was smaller at 71 days of age in the treatment groups. A decrease in estradiol levels for each group was observed at 50 days of age, but not at later time points. Progesterone levels were reduced in the 4-HPR group, but not in the dietary-restricted group, during each time period. It would appear that the observed decrease in mammary cancers observed with moderate reductions in body weight gain might be due to multiple related factors different from those related to 4-HPR treatment.