Dying produces human remains and is a unique period of human lives that remains poorly understood. The aims of this research were to validate the use of hair cortisol concentration (HCC) analysis in the study of stress at the end of life and to explore the effects of biocultural factors on stress experience in the last months of life. This study examined the dead from the 1st century CE Egypt, 19th-20th century Missouri, and 21st century Florida. A framework of embodiment and the good death was employed to interpret lived experience from HCC and examine the relationships between HCC, death, cause of death, duration of disease, and medical care and treatment. HCC in the dead is higher than in the living, varies considerably between individuals, and can fluctuate across the last months of life. High HCC at the end of life cannot be easily accounted for by medications, serious disease, or decomposition; are within possible biological ranges; and are dynamic. However, leaching of cortisol from the archaeological samples is likely. Duration of disease or presence of multiple medical conditions does not influence HCC. While modern medical advancements do not improve stress levels in the dead, a reduction in stress is observed following hospital entry in the past. HCC is a valid measure of stress at the end of life. The last months of life are periods of significant stress but dying is an inherently personal and dynamic experience that varies between individuals and over many months leading up to death due to the interaction of multiple biocultural factors. These findings contribute to the understanding of a unique period of individual lives, suggest that studies of HCC in bioarchaeology must focus on the dying period and be wary of leaching, and highlight the potential of HCC in palliative care research. / Dissertation / Doctor of Philosophy (PhD) / Today, death is often regarded with uncertainty and even fear, yet little is known about the experience of dying, especially in the past. Dying is difficult to study in modern people because of communication challenges and the number of complex factors at play while studies of the past are limited because human remains do not reveal how individuals felt. A better understanding of the dying experience can help bioarchaeologists clarify the relationship between dying, death, and skeletal remains and could help improve the care of dying people today. This research evaluated a new method of stress assessment to study experiences in the last months of life in archaeological, historic, and modern samples. Hair cortisol concentration (HCC) analysis measures stress hormones in human hair to reconstruct stress experience at the time the hair was growing. High levels of HCC in dead individuals from 1st century AD Egypt, 19th-20th century Missouri, and 21st century Florida indicate that dying is stressful across time and place. HCC values from the Egyptian sample were higher than most living people but were lower than other archaeological samples which may be a result of cortisol leaching out of the hair shaft after death. Although higher than living people, HCC levels still differed between individuals and across individual hair shafts, indicating that stress experience can be different between individuals and change over an individual’s final months. These differences are not a result of biological factors such as duration of disease or the presence of certain preexisting conditions suggesting that stress at the end of life is complex. Despite advancements in medicine, the modern sample displayed similar HCC levels to those from earlier historic periods and for a number of individuals from the historic sample, hospital entry led to a temporary reduction in HCC levels. Together, these findings suggest that, while modern medical advancements have not improved stress levels at the end of life, some aspects of care could reduce stress. The results of this study indicate that dying is a stressful, complex, and dynamic phenomenon that modern medical treatment alone may not be able to improve. Furthermore, studies of HCC in archaeology must focus on the effects of dying and be wary of leaching. Ultimately, HCC analysis could contribute to a greater knowledge of the dying experience, the understanding of past peoples, and improvement of the experience of dying.

Cortisol is a stress hormone which is released from the adrenals in the Hypothalamic-Pituitary-Adrenal (HPA) axis and plays a major role in animal stress response. Cortisol is used as a stress marker and can be sampled using different methods. A good non-invasive method and a good measure of chronic stress is to measure cortisol through hair. Cortisol is stored in hair for months and therefore reflects chronic stress. The aim of this study was to investigate if cortisol concentration differs depending on coat color. Hair samples from 20 black and white Border collie dogs was analysed and used in this study. Cortisol was extracted with methanol and analysed with ELISA. The results showed no significant difference between black and white coat color within the population while there were individual differences. The results also showed that the sexes do not affect the cortisol concentration. In summary, coat color (black and white) has an effect on cortisol concentration which means that the factor color does need to be taken into account when measuring cortisol through hair.

Prenatal stress has been correlated with adverse developmental outcomes affecting infant cognition and behavior. Previous studies have shown that prenatal stress can lead to increased susceptibility to adult disease but few studies have looked at the physiological stress response system by measuring the activity of the hypothalamicpituitary-adrenal (HPA) axis. Cortisol, the output of the HPA axis can be secreted in many different matrices (saliva, blood, urine, feces and hair). Most studies that do, only look at one measure of hormone production instead of examining multiple matrices. Additionally these studies do not look at the relationship between matrices. Hair provides a long-term assessment of cortisol hormone production as related to infant behavior. Four measures of cortisol representative of prenatal and postpartum periods were collected in a sample population of rhesus macaques at the NIH facility. No stress was applied to these animals and cortisol concentrations were assessed in maternal hair, infant hair, amniotic fluid, and mothers’ milk. These cortisol measures were then analyzed first to determine vii the relationships between the four measures and second to relate these cortisol values to infant behavior in the primate neonatal neurobehavioral assessment. Subjects of this study were 30 mothers and infants from the 2015 and 2016 breeding cohort. 25 of which, were unique dyads. Using four statistical analyses and 3 groupings of behavior, we found that maternal hair cortisol concentrations were correlated with different temperaments of infants, while milk cortisol concentrations were correlated with infant’s visual exploration of the environment. Additionally, an inverse relationship was found between hair cortisol concentrations and both hair cortisol concentrations with amniotic fluid cortisol. Together, the four statistical analyses show that Maternal HPA axis activation during and after pregnancy affects infant behavioral development 1 month postpartum.

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<p>Dairy calves in the preweaning period are exposed to a variety of stressors (e.g. disbudding, castration, etc.) and management practices that have the potential to influence their health and productivity later in life. Proper colostrum management at birth is the first step in ensuring calves can reach their full potential in terms of health, growth, and development, while improper management increases a calf’s risk of failed transfer of passive immunity <strong>(FTPI)</strong> which puts the calf at an increased risk of morbidity and mortality. Previous research has indicated there is a difference in colostrum management practices between dairy bull and heifer calves. Due to the marketing of dairy-beef crossbred calves as surplus animals along with dairy bull calves, there is growing interest in the treatment of these calves on farms. The first study presented here (Chapter 2) investigated the relationship between colostrum management practices on 15 farms and the prevalence of FTPI between purebred dairy and dairy-beef crossbred calves on those farms. The results of this study found no influence of calf breed (purebred dairy or dairy-beef crossbred) on the prevalence of FTPI. However, farm tended to influence FTPI. There was also a significant influence of the reported timing of the first colostrum feeding on FTPI, with calves on farms reporting feeding colostrum within 1 hour of birth having decreased odds of experiencing FTPI. In addition to colostrum management at birth, calves are exposed to several management factors during the preweaning period that can cause stressful experiences and result in negative behavioral and physiological outcomes. Early life stress, especially chronic stress, can negatively impact calves later in life. Short term stress load in calves can be quantified by measuring cortisol in the blood serum or saliva; however, these measures are not reflective of chronic stress load because they only measure circulating cortisol. The second study (Chapter 3) considered the efficacy of utilizing hair cortisol concentration <strong>(HCC)</strong> as a measure of stress load in 27 dairy calves exposed to a weekly saline injection or a weekly or biweekly repeated acute stressor (Cosyntropin injection) during the preweaning period. The results revealed no difference in HCC between the treatment groups, though salivary cortisol was increased in calves injected with Cosyntropin. Additionally, calf behavior and growth were not influenced by the treatments in this study. Overall, these studies provided insights into two areas that have not been extensively researched. First, considering dairy-beef crossbred calf management, and second investigating a method to assess chronic stress that has not been extensively researched in calves.</p>

Cognitive impairment in coronary artery disease (CAD) patients can predict poorer quality of life, dementia, and increased mortality. This study aimed to determine the association between long-term cortisol elevations and cognitive function in CAD patients. Participants were recruited at the beginning of a 1 year cardiac rehabilitation program and followed forward. Composite Z-scores were computed from tests measuring memory and executive function at baseline and 1 year. Cortisol deposition (3 months) was measured from a 20 mg, 3 cm hair sample. Analyses of covariance showed less improvement in memory function (F1,50=4.721, p=0.035) but not executive function (F1,49=0.318, p=0.575) in patients dichotomized into a high cortisol group based on a previously established reference range. Prolonged cortisol elevation may be associated with cognitive changes in subjects with CAD.

Cognitive impairment in coronary artery disease (CAD) patients can predict poorer quality of life, dementia, and increased mortality. This study aimed to determine the association between long-term cortisol elevations and cognitive function in CAD patients. Participants were recruited at the beginning of a 1 year cardiac rehabilitation program and followed forward. Composite Z-scores were computed from tests measuring memory and executive function at baseline and 1 year. Cortisol deposition (3 months) was measured from a 20 mg, 3 cm hair sample. Analyses of covariance showed less improvement in memory function (F1,50=4.721, p=0.035) but not executive function (F1,49=0.318, p=0.575) in patients dichotomized into a high cortisol group based on a previously established reference range. Prolonged cortisol elevation may be associated with cognitive changes in subjects with CAD.

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Background: The effect of animal-assisted activities (AAA) on the animal participants has been minimally investigated and the welfare of these animals has been questioned. Cortisol, in conjunction with stress-associated behavior, has been utilized as an objective assessment of animal welfare. Objective: Salivary cortisol and behavior in AAA dogs were measured to test the null hypothesis that salivary cortisol concentration and behavior are not different in an AAA environment compared to home or neutral environments.  Hair cortisol was measured in AAA dogs to test the null hypothesis that there is no relationship between hair cortisol and salivary cortisol. Methods:  Fifteen healthy adult dogs registered with an AAA organization were recruited.  A hair sample was collected from each dog upon enrollment.  Saliva samples were collected from each dog every 30 minutes, starting 30 minutes prior to and 30 minutes after a standardized 60 minute session across 3 settings: an AAA session (AS) for college students in the communal area of a residence hall; a neutral session (NS) located in a novel room without interaction with a stranger; and a home session (HS).  Each session was videotaped continuously and behaviors were coded at three separate 5-minute intervals while the dog was petted by a stranger in the AS or handler in the NS and HS. Results:  Salivary cortisol levels were not different in the AS compared to HS, but were significantly higher in the NS compared to AS and HS.  Dogs exhibited significantly more standing and ambulating behavior in the AS compared to HS.  Salivary cortisol level was negatively correlated with panting and standing at specific time points in the NS and AS, respectively.  Hair cortisol level did not correlate with salivary cortisol level at any time point in any of the settings. Conclusions:  During a 60 minute AAA session, salivary cortisol concentration and stress-associated behavior were not different compared to when dogs spent the same amount of time in the home setting, suggesting that they were not stressed when being used as AAA animals. The physical environment may be an important consideration when evaluating the effect of AAA on dogs.  Hair cortisol did not correlate with salivary cortisol, suggesting that hair may not be a representative predictor of cortisol in these environments.  Additional investigation is required to support cortisol and behavior as measures of stress and welfare in AAA animals. / Master of Science