Lead exposure across the life course and age at death.

BACKGROUND: Lead (Pb) exposure has been associated with an increased risk of all-cause mortality, even at low levels. Little is known about how the timing of Pb exposure throughout life may influence these relationships. Quantifying the amount of Pb present in various tissues of the body provides measurements of exposure from different periods of life. These include bone, tooth enamel, which is the hard outer layer of the crown, and tooth cementum, which is the calcified connective tissue covering the tooth root. The purpose of the study was to examine Pb exposure at multiple periods throughout life, including childhood (enamel), adulthood (cementum), and later life (bone), and to estimate their associations with age at death. METHODS: 208 skeleton donors (born 1910-1960) from an ongoing case-control study were included in this study. Pb was measured in tibia (shin), bone using X-Ray Florescence and in teeth using Laser-Ablation Inductively Coupled Plasma Mass Spectroscopy. After excluding unusually high measurements (>2sd), this resulted in a final sample of 111 with all exposure measures. Correlations across measures were determined using partial Spearman correlations. Associations between Pb exposure and age at death were estimated using Multivariable Linear Regression. RESULTS: Pb measures across exposure periods were all significantly correlated, with the highest correlation between cementum and tibia measures (r = 0.61). Donors were largely female (63.0 %), White (97.3 %), and attended some college (49.5 %). Single exposure models found that higher tooth cementum Pb (-1.27; 95 % CI: -2.48, -0.06) and tibia bone Pb (-0.91; 95 % CI: -1.67, -0.15) were significantly associated with an earlier age at death. When considered simultaneously, only cementum Pb remained significant (-1.51; 95 % CI: -2.92, -0.11). Secondary analyses suggest that the outer cementum Pb may be especially associated with an earlier age at death. CONCLUSION: Results suggest that higher Pb exposure is associated with an earlier age at death, with adulthood as the life period of most relevance. Additional studies using Pb exposure measures from different life stages should be conducted.

Soil elemental changes during human decomposition.

Mammalian decomposition provides pulses of organic matter to the local ecosystem creating ephemeral hotspots of nutrient cycling. While changes to soil biogeochemistry in these hotspots have been described for C and N, patterns associated with deposition and cycling of other elements have not received the same attention. The goal of our study was to evaluate temporal changes to a broad suite of dissolved elements in soils impacted by human decomposition on the soil surface including: 1) abundant mineral elements in the human body (K, Na, S, P, Ca, and Mg), 2) trace elements in the human body (Fe, Mn, Se, Zn, Cu, Co, and B), and 3) Al which is transient in the human body but common in soils. We performed a four-month human decomposition trial at the University of Tennessee Anthropology Research Facility and quantified elemental concentrations dissolved in the soil solution, targeting the mobile and bioavailable fraction. We identified three groups of elements based on their temporal patterns. Group 1 elements appeared to be cadaver-derived (Na, K, P, S) and their persistence in soil varied based upon soluble organic forms (P), the dynamics of the soil exchange complex (Na, K), and gradual releases attributable to microbial degradation (S). Group 2 elements (Ca, Mg, Mn, Se, B) included three elements that have greater concentrations in soil than would be expected based on cadaver inputs alone, suggesting that these elements partially originate from the soil exchange (Ca, Mg), or are solubilized as a result of soil acidification (Mn). Group 3 elements (Fe, Cu, Zn, Co, Al) increased late in the decomposition process, suggesting a gradual solubilization from soil minerals under acidic pH conditions. This work presents a detailed longitudinal characterization of changes in dissolved soil elements during human decomposition furthering our understanding of elemental deposition and cycling in these environments.

Bone lead variability in bone repository skeletal samples measured with portable x-ray fluorescence.

Bone lead serves as a better, more accessible biomarker to many communities experiencing chronic exposure to lead. A new method using low energy x-ray fluorescence in a handheld device (portable XRF) allows us to measure this chronic biomarker in only a few minutes. However, many unknowns remain about this biomarker measured using a new low energy x-ray technique. The low energy of the new method was theorized to measure a slightly different portion of the bone than previous techniques, which could influence measurements at different bone sites and types. We tested how bone measurements varied across five bone sites: mid-tibial shaft, proximal tibia, distal tibia (ankle), ilium, and cranium. We found bone lead measurements are not significantly different between skeletal elements when measured using a portable XRF. On average, bone lead in the repository samples was measured to be 21.6 ± 21.3 µg/g with an XRF detection limit of 2.1 ± 0.5 µg/g. Cumulative lead exposure can be effectively measured using the portable XRF on a variety of bone types, but the tibia should be preferentially measured to compare between studies and individuals.

Not by the Book: Observations of Delayed Oviposition and Re-Colonization of Human Remains by Blow Flies.

Postmortem interval estimations can be complicated by the inter-individual variation present in human decomposition. Forensic entomologists may especially face challenges interpreting arthropod evidence in scenarios that are not "by the book", or that vary in unexpected ways. Therefore, it is important to report instances where blow fly colonization does not align with expected soft tissue decomposition as blow fly larvae are often used to produce a time of colonization (TOC) estimation to infer a minimum PMI. We followed the decomposition and blow fly activity of three human donors at the Anthropology Research Facility (University of Tennessee). Delayed oviposition occurred on one donor 115 d post-placement, whereas two donors experienced blow fly re-colonization after cessation of the consumption phase, one 22 d and one more than 200 d after blow fly larvae were last observed. A null hypothesis model tested whether the entomological TOC and anthropological total body score (TBS) estimations encompassed the time of placement (TOP) for each donor. While the null hypothesis was rejected for all TOC estimations, it could not be rejected for the TBS estimations. We discuss how the non-linear nature of human decomposition can pose challenges to interpreting blow fly evidence and suggest that forensic entomology practitioners should recognize these limitations in both research endeavors and applied casework.

Body Mass Index (BMI) Impacts Soil Chemical and Microbial Response to Human Decomposition.

Microorganisms are key decomposers of vertebrate mortalities, breaking down body tissues and impacting decomposition progress. During human decomposition, both extrinsic environmental factors and intrinsic cadaver-related factors have the potential to impact microbial decomposers either directly or indirectly via altered physical or chemical conditions. While extrinsic factors (e.g., temperature, humidity) explain some variation in microbial response during human decomposition in terrestrial settings, recent work has noted that even under the same environmental conditions, individuals can have different decomposition patterns, highlighting the potential for intrinsic factors to impact microbial decomposers. The goal of this study was to investigate the effects of several intrinsic factors (age, sex, diseases at time of death, and body mass index [BMI]) on chemical and microbial changes in decomposition-impacted soils. In a field study conducted at the University of Tennessee Anthropology Research Facility, soils were collected from the decomposition-impacted area surrounding 19 deceased human individuals through the end of active decomposition. Soil physicochemical parameters were measured, and microbial (bacterial and fungal) communities were assessed via amplicon sequencing. BMI was shown to explain some variation in soil pH and microbial response to human decomposition. Hierarchical linear mixed (HLM) effects models revealed that BMI category significantly explained variation in pH response within decomposition-impacted soils over time (HLM F = 9.647; P < 0.001). Additionally, the relative abundance of soil Saccharomycetes in decomposition soils under underweight donors displayed little to no changes (mean maximum change in relative abundance, +6.6%), while all other BMI categories displayed an increased relative abundance of these organisms over time (normal, +50.6%; overweight, +64.4%; and obese, +64.6%) (HLM F = 3.441; P = 0.11). Together, these results reveal intrinsic factors influencing decomposition patterns, especially within the soil environment, and suggest BMI is an important factor for controlling decomposition processes. IMPORTANCE This work begins to address questions about interindividual variation in vertebrate decomposition attributed to intrinsic factors, that is, properties of the carcass or cadaver itself. Most research on factors affecting decomposition has focused on the extrinsic environment, such as temperature or humidity. While these extrinsic factors do explain some variation in decomposition patterns, interindividual variability is still observed. Understanding how intrinsic factors influence microbial decomposers will help reveal the ecological impacts of decomposition. This work also has forensic applications, as soil chemical and biological changes have been suggested as indicators of postmortem interval. We reveal factors that explain variation in the decomposition environment that should be considered in these estimates. This is particularly important as we consider the implications of variations in human populations due to diet, age, BMI, disease, toxicological loading, etc. on forensic investigations dealing with decomposing remains.

First record of the oriental latrine fly, Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae), in Tennessee, USA.

PURPOSE: Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) is a medically and forensically important blow fly species that invaded the United States three decades ago and has continued to expand its distribution across the country ever since. Unlike many other blow fly species, larvae of C. megacephala can develop in feces, particularly from humans. Additionally, C. megacephala is known to carry antibiotic-resistant bacteria in even greater quantities than house flies in tropical areas. This behavior, along with its dissemination into and prevalence within human inhabited environments (e.g., outdoor markets, urban neighborhoods), makes C. megacephala a potential threat to human health. This short communication serves as the first record of C. megacephala in Tennessee, USA. METHODS: Collections of adult and larval blow flies were made from two sets of human remains decomposing at the Anthropology Research Facility (ARF) at the University of Tennessee. Specimens were confirmed by an expert to be C. megacephala. RESULTS: A total of seven individual specimens (four adults and three larvae) of C. megacephala were collected from human bodies at the ARF. These results indicate that C. megacephala is not just dispersing into this environment as adults, but actively colonizing human remains in this semi-urban area. CONCLUSION: These observations support the previously described behavior and habitat of this filth-breeding fly in many Asian countries where it is considered medically and forensically important. Therefore, continuously updated distribution records, like this one, are critical for tracking the movement of C. megacephala across the United States.

Plants to Remotely Detect Human Decomposition?

In the USA, 100 000 people go missing every year. Difficulty in the rapid identification of sites of human decomposition complicates the recovery of bodies, especially in forests. We propose that spectral responses in tree and shrub canopies could act as guides to find cadavers using remote sensing platforms for societal benefit.

Comparative Decomposition of Humans and Pigs: Soil Biogeochemistry, Microbial Activity and Metabolomic Profiles.

Vertebrate decomposition processes have important ecological implications and, in the case of human decomposition, forensic applications. Animals, especially domestic pigs (Sus scrofa), are frequently used as human analogs in forensic decomposition studies. However, recent research shows that humans and pigs do not necessarily decompose in the same manner, with differences in decomposition rates, patterns, and scavenging. The objective of our study was to extend these observations and determine if human and pig decomposition in terrestrial settings have different local impacts on soil biogeochemistry and microbial activity. In two seasonal trials (summer and winter), we simultaneously placed replicate human donors and pig carcasses on the soil surface and allowed them to decompose. In both human and pig decomposition-impacted soils, we observed elevated microbial respiration, protease activity, and ammonium, indicative of enhanced microbial ammonification and limited nitrification in soil during soft tissue decomposition. Soil respiration was comparable between summer and winter, indicating similar microbial activity; however, the magnitude of the pulse of decomposition products was greater in the summer. Using untargeted metabolomics and lipidomics approaches, we identified 38 metabolites and 54 lipids that were elevated in both human and pig decomposition-impacted soils. The most frequently detected metabolites were anthranilate, creatine, 5-hydroxyindoleacetic acid, taurine, xanthine, N-acetylglutamine, acetyllysine, and sedoheptulose 1/7-phosphate; the most frequently detected lipids were phosphatidylethanolamine and monogalactosyldiacylglycerol. Decomposition soils were also significantly enriched in metabolites belonging to amino acid metabolic pathways and the TCA cycle. Comparing humans and pigs, we noted several differences in soil biogeochemical responses. Soils under humans decreased in pH as decomposition progressed, while under pigs, soil pH increased. Additionally, under pigs we observed significantly higher ammonium and protease activities compared to humans. We identified several metabolites that were elevated in human decomposition soil compared to pig decomposition soil, including 2-oxo-4-methylthiobutanoate, sn-glycerol 3-phosphate, and tryptophan, suggesting different decomposition chemistries and timing between the two species. Together, our work shows that human and pig decomposition differ in terms of their impacts on soil biogeochemistry and microbial decomposer activities, adding to our understanding of decomposition ecology and informing the use of non-human models in forensic research.

The Influence of Body Size on the Expression of Sexually Dimorphic Morphological Traits.

Skeletal sexual dimorphism manifests as size or shape differences between males and females in a population. Certain dimorphic traits are used in sex estimation methods, and populational variation in the expression of these traits can result in inaccurate sex estimation. However, the underlying causes of variation in trait expression remain unclear. This study explores body size, which also exhibits sexual dimorphism, as a potential factor influencing trait expression. To test this, skeletons of 209 individuals of varying body size were analyzed, and morphological traits were scored according to the Walker (2008), Klales et al. (2012), and Rogers (1999) sex estimation methods. Statistical analyses found significant correlations between body size parameters and expression of traits, with stature explaining more relative variance in trait expression than body mass. However, the relationships are weak and few in number, suggesting that body size has a minimal impact on the expression of these morphological traits.

A study of the intrinsic variability and the effect of environmental conditions on the formation of a postmortem root band.

A postmortem root band (PMRB) is defined as "an opaque ellipsoidal band composed of a collection of parallel elongated air/gas spaces and is approximately 0.5mm above the root bulb and about 2mm below the skin surface" [1]. It is generally accepted that it can appear in the root of hairs attached to remains during decomposition [1]. This study aimed to investigate the underlying cause and mechanism of PMRB formation. This was done (i) by observing the overall frequency and the intrinsic variability in anagen hairs containing a PMRB collected across five regions of a human decedent's scalp at three time points, and (ii) by determining if PMRB-like features can be induced via immersion in in-vitro controlled environments of anagen hairs plucked from the scalp of a human decedent (ex-situ postmortem hairs) not containing a PMRB. The results of the first objective illustrated that as time since death increased, the frequency of hairs containing a PMRB across the scalp sampling regions increased and the intrinsic variability decreased. The results of the second objective demonstrated that both an aqueous environment and microbial activity are essential for the formation of PMRB-like features. This study was the first to statistically analyze the intrinsic variability of PMRB formation, as well as the first to induce PMRB-like features in roots of ex-situ postmortem hairs.

Impact of sample degradation and inhibition on field-based DNA identification of human remains.

The advent of DNA technologies for field-based application promises to provide rapid intelligence to aid investigations. Their validation and adoption by enforcement groups have demonstrated utility in sample screening and prioritisation, but field application in some areas of forensic science, such as human remains identification, is little evidenced. Assessing the ability of such approaches to provide meaningful data is critical as decomposition is likely to complicate analysis and limit the effective use of such field-based DNA interventions. This research assessed the ability to collect viable DNA data in the field using the ParaDNA Field Instrument and Intelligence Test chemistry. Different sample collection methods were assessed; direct from skin surface; direct from exposed tissue; indirect from muscle swab transferred to FTA card; and from larvae on the donors. Samples were collected and processed on-site at the Anthropology Research Facility, University of Tennessee. The data show that the muscle tissue provided the most effective sample template and, using this approach, it was possible to generate STR profiles from human remains in under two hours from the time of sample collection. STR profile data were collected up to four days from donor placement (114 Accumulated Degree Days). After this time there was a rapid decrease in the quality of the profiles collected due to the onset of decomposition. The data also show that effective sample recovery was not possible from the surface of the skin, exposed tissue or from carrion larvae. Inhibition studies in the laboratory suggest that by-products of the decomposition process are the primary mode of failure. Together these data suggests a possible application for screening and prioritisation in criminal casework but highlights issues that may affect the success of the approach.

Selective mortality in middle-aged American women with Diffuse Idiopathic Skeletal Hyperostosis (DISH).

OBJECTIVE: A mortality sample of white American male and female skeletons was examined to illustrate a simple means of identifying skeletal conditions associated with an increased risk of dying relatively early in adulthood and to determine if males and females with Diffuse Idiopathic Skeletal Hyperostosis (DISH) displayed the same general age-specific pattern of mortality. METHODS: Age-specific probability distributions for DISH were generated from 416 white Americans who died from the 1980s to the present, and whose remains were donated to the University of Tennessee Forensic Anthropology Center. The age-specific frequency of DISH is analyzed using an empirical smoothing algorithm. Doing so allows for the identification of deviations (i.e., local maxima) from monotonically increasing age-specific probabilities. RESULTS: In females (N = 199), there is a peak in the frequency of individuals with DISH around 60 years of age where 37.0% of the individuals have DISH. It is matched only by the frequency (38.7%) in the oldest females, those over 85 years old. In contrast, DISH frequencies for males (N = 217) increase monotonically with advancing age, reaching 62.5% in the ≥86 years age group. There was an association between DISH and high body weight in women, particularly those who died before they reached the age of 75. CONCLUSIONS: Early-onset DISH in white American women is associated with an increased risk of dying indicated by a local maximum in the probability curve. Should this finding be replicated in additional mortality samples and the reason DISH is associated with early death is established, beyond being heavy, this radiologically visible ossification of the spine could be a potential component of health-monitoring programs for middle-aged women.

Age-at-Death Estimation for Modern Populations in Mexico and Puerto Rico through the Use of 3D Laser Scans of the Pubic Symphysis.

Reliable age-at-death estimates from the adult skeleton are of fundamental importance in forensic anthropology, because it contributes to the identity parameters used in a medicolegal death investigation. However, reliable estimates are difficult because many traditional aging methods depend on a set of population-specific criteria derived from individuals of European and African descent. The absence of information on the potential differences in the aging patterns of underrepresented, especially Latinx, populations may hinder our efforts to produce useful age-at-death estimates. In response to these concerns, this study explores the utility of currently available aging techniques and whether population-specific aging methods among Latinx groups are needed. The authors obtained data from two skeletal collections representing modern individuals of Mexican and Puerto Rican origin. They examined five newly developed computational shape-based techniques using 3D laser scans of the pubic symphysis and one traditional bone-to-phase technique. A validation test of all computational and traditional methods was implemented, and new population-specific equations using the computational algorithms were generated and tested against a subsample. Results suggest that traditional and computational aging techniques applied to the pubic symphysis perform best with individuals within 35-45 years of age. Levels of bias and inaccuracy increase as chronological age increases, with overestimation of individuals younger than 35 years and underestimation of individuals older than 45 years. New regression models provided error rates comparable to, and in some occasions outperformed, the original computational models developed on white American males, but age estimates did not significantly improve. This study shows that population-specific models do not necessarily improve age estimates in Latinx samples. Results do suggest that computational methods can ultimately outperform the Suchey-Brooks method and provide improved objectivity when estimating age at death in Latinx samples.