Dispatches from the interface of salivary bioscience and neonatal research.

The emergence of the interdisciplinary field of salivary bioscience has created opportunity for neonatal researchers to measure multiple components of biological systems non-invasively in oral fluids. The implications are profound and potentially high impact. From a single oral fluid specimen, information can be obtained about a vast array of biological systems (e.g., endocrine, immune, autonomic nervous system) and the genetic polymorphisms related to individual differences in their function. The purpose of this review is to describe the state of the art for investigators interested in integrating these unique measurement tools into the current and next generation of research on gonadal steroid exposure during the prenatal and neonatal developmental periods

Uric Acid as a Potential Peripheral Biomarker for Disease Features in Huntington's Patients.

Oxidative stress has long been implicated in the pathophysiology and progression of Huntington's disease (HD). Uric acid (UA) is a naturally occurring antioxidant that is present in the brain and periphery. Growing evidence has implicated UA as a molecular biomarker for several neurodegenerative diseases, most notably Parkinson's disease (PD). In this study, we investigated UA levels in clinical samples from HD patients and normal controls (NCs) and assessed potential relationships between UA levels and disease and clinical data. UA levels were measured in plasma (n = 107) and saliva (n = 178) samples from premanifest (pre-HD) and manifest HD patients and control subjects. Gender effects of UA levels were observed in both biofluids, with male patients showing higher UA levels compared to female patients. Comparisons of UA levels across diagnostic groups, separated by gender, revealed that both plasma and salivary UA levels were significantly lower in female pre-HD and manifest HD patients compared to NCs. Salivary levels of UA were also significantly lower in male manifest HD patients versus controls, but not in plasma. Correlations of peripheral UA levels to clinical data also showed differences according to gender. In male HD patients, both plasma and salivary UA levels were significantly negatively correlated with total functional capacity (TFC), while positive correlations were observed with total motor score (TMS). Female HD patients showed a significant positive correlation between plasma UA levels and TMS, while salivary UA levels from female patients were significantly correlated to disease burden. Finally, in a separate cohort, we show that UA levels are decreased in postmortem prefrontal cortical samples (n = 20) from HD subjects compared to matched controls. These findings suggest that decreased levels of UA in the brains of HD patients can be reflected in peripheral fluids, with salivary measures of UA particularly offering significant promise as a potentially relevant, non-invasive biomarker of disease symptoms and burden. Our findings further highlight the impact of sexual dimorphism in HD pathophysiology

Correspondence Between Cytomegalovirus Immunoglobulin-G Levels Measured in Saliva and Serum

Human cytomegalovirus (HCMV) infects more than 80% of the global population. While mostly asymptomatic, HCMV infection can be serious among the immunocompromised, and it is implicated in chronic disease pathophysiology in adulthood. Large-scale minimally invasive HCMV screening could advance research and public health efforts to monitor infection prevalence and prevent or mitigate downstream risks associated with infection. We examine the utility of measuring HCMV immunoglobulin-G (IgG) levels in saliva as an index of serum levels. Matched serum and saliva samples from healthy adults (N = 98; 44% female; 51% white) were assayed for HCMV IgG, total salivary protein, and salivary markers related to oral inflammation, blood, and tissue integrity. We examine the serum-saliva association for HCMV IgG and assess the influence of participant characteristics and factors specific to the oral compartment (e.g., oral inflammation) on HCMV IgG levels and cross-specimen relations. We found a robust serum-saliva association for HCMV IgG with serum antibody levels accounting for \u3e60% of the variance in salivary levels. This relation remained after adjusting for key demographic and oral immune-related variables. Compared to the serum test, the salivary HCMV IgG test had 51% sensitivity and 97% specificity. With improvements in assay performance and sample optimization, HCMV antibody levels in oral fluids may be a useful proxy for serum levels

Salivary total Immunoglobulin G as a surrogate marker of oral immune activity in salivary bioscience research

The integration of salivary biomeasures in biobehavioral, psychophysiological, and clinical research has greatly expanded our ability to study the biopsychosocial processes underlying health. Much of this research, however, has failed to adequately assess and adjust for the impact of oral immune activity on salivary biomeasure concentrations and associations with serum levels. Aiming to improve the validity and reliability of salivary biomeasure data, we examine salivary total Immunoglobulin G (IgG) as a potential surrogate marker of oral inflammation and immune activity. During a single study visit in Baltimore, Maryland, healthy young adult participants provided matched blood and saliva samples (N=99; age 18–37 years, 42% female) and completed an oral health questionnaire. Biospecimens were assayed for total IgG and immune markers related to inflammation (cytokines), blood in saliva (transferrin), and tissue remodeling (matrix metalloproteinase-8). Total IgG (μg/mL) concentrations were higher in serum than saliva. Salivary total IgG was associated with some self-reported oral health measures, and strongly positively associated with all salivary immune markers. Controlling for salivary total IgG may be a feasible, affordable approach to adjusting salivary biomeasure findings for the influence of the oral immune environment when it is not possible or practical to obtain clinical oral health data

Censored data considerations and analytical approaches for salivary bioscience data

Left censoring in salivary bioscience data occurs when salivary analyte determinations fall below the lower limit of an assay’s measurement range. Conventional statistical approaches for addressing censored values (i.e., recoding as missing, substituting or extrapolating values) may introduce systematic bias. While specialized censored data statistical approaches (i.e., Maximum Likelihood Estimation, Regression on Ordered Statistics, Kaplan-Meier, and general Tobit regression) are available, these methods are rarely implemented in biobehavioral studies that examine salivary biomeasures, and their application to salivary data analysis may be hindered by their sensitivity to skewed data distributions, outliers, and sample size. This study compares descriptive statistics, correlation coefficients, and regression parameter estimates generated via conventional and specialized censored data approaches using salivary C-reactive protein data. We assess differences in statistical estimates across approach and across two levels of censoring (9% and 15%) and examine the sensitivity of our results to sample size. Overall, findings were similar across conventional and censored data approaches, but the implementation of specialized censored data approaches was more efficient (i.e., required little manipulations to the raw analyte data) and appropriate. Based on our review of the findings, we outline preliminary recommendations to enable investigators to more efficiently and effectively reduce statistical bias when working with left-censored salivary biomeasure data

Overestimating Self-Blame for Stressful Life Events and Adolescents’ Latent Trait Cortisol (LTC): The Moderating Role of Parental Warmth. Journal of Youth and Adolescence

Cognitive interpretations of stressful events impact their implications for physiological stress processes. However, whether such interpretations are related to trait cortisol—an indicator of individual differences in stress physiology—is unknown. In 112 early adolescent girls (M age = 12.39 years), this study examined the association between self-blame estimates for past year events and latent trait cortisol, and whether maternal warmth moderated effects. Overestimating self-blame (versus objective indices) for independent (uncontrollable) events was associated with lower latent trait cortisol, and maternal warmth moderated the effect of self-blame estimates on latent trait cortisol for each dependent (at least partially controllable) and interpersonal events. Implications for understanding the impact of cognitive and interpersonal factors on trait cortisol during early adolescence are discussed

Salivary Cortisol Mediates Effects of Poverty and Parenting on Executive Functions in Early Childhood

In a predominantly low-income population-based longitudinal sample of 1,292 children followed from birth, higher level of salivary cortisol assessed at ages 7, 15, and 24 months was uniquely associated with lower executive function ability and to a lesser extent IQ at age 3 years. Measures of positive and negative aspects of parenting and household risk were also uniquely related to both executive functions and IQ. The effect of positive parenting on executive functions was partially mediated through cortisol. Typical or resting level of cortisol was increased in African American relative to White participants. In combination with positive and negative parenting and household risk, cortisol mediated effects of African American ethnicity, income-to-need, and maternal education on child cognitive ability.

Oral microbial communities in children, caregivers, and associations with salivary biomeasures and environmental tobacco smoke exposure

Human oral microbial communities are diverse, with implications for oral and systemic health. Oral microbial communities change over time; thus, it is important to understand how healthy versus dysbiotic oral microbiomes differ, especially within and between families. There is also a need to understand how the oral microbiome composition is changed within an individual including by factors such as environmental tobacco smoke (ETS) exposure, metabolic regulation, inflammation, and antioxidant potential. Using archived saliva samples collected from caregivers and children during a 90-month follow-up assessment in a longitudinal study of child development in the context of rural poverty, we used 16S rRNA gene sequencing to determine the salivary microbiome. A total of 724 saliva samples were available, 448 of which were from caregiver/child dyads, an additional 70 from children and 206 from adults. We compared children’s and caregivers’ oral microbiomes, performed “stomatotype” analyses, and examined microbial relations with concentrations of salivary markers associated with ETS exposure, metabolic regulation, inflammation, and antioxidant potential (i.e., salivary cotinine, adiponectin, C-reactive protein, and uric acid) assayed from the same biospecimens. Our results indicate that children and caregivers share much of their oral microbiome diversity, but there are distinct differences. Microbiomes from intrafamily individuals are more similar than microbiomes from nonfamily individuals, with child/caregiver dyad explaining 52% of overall microbial variation. Notably, children harbor fewer potential pathogens than caregivers, and participants’ microbiomes clustered into two groups, with major differences being driven by Streptococcus spp. Differences in salivary microbiome composition associated with ETS exposure, and taxa associated with salivary analytes representing potential associations between antioxidant potential, metabolic regulation, and the oral microbiome

Emotion Regulation and Positive Affect in the Context of Salivary Alpha-Amylase Response to Pain in Children with Cancer: Physiology, Self-Report, and Behavior

Children with cancer are repeatedly exposed to aversive stimuli including painful procedures. Therefore, emotional regulation techniques may prove useful during such experiences and contribute to pain resilience. This study aimed to determine whether three different emotional regulation strategies (distraction, reappraisal and reassurance) impacted physiological, self- reported and behavioral pain responses in pediatric patients with cancer ages 6 to 18 years (N = 73). The cold pressor task (CPT), an experimental task in which pain is induced by having participants place their hand in cold water, was used to examine pain responses. Patients placed their hand in 7 degree Celsius water for up to 4 minutes. Saliva samples were collected 15 minutes before, immediately after, and then 15 minutes after the CPT. Saliva samples were assayed for alpha amylase, a proxy for sympathetic nervous system activation. Self-reported pain severity was measured upon hand removal. Pain tolerance was assessed by length of time participants kept their hand in the water. Children in the reassurance condition exhibited salivary alpha amylase levels that continued to rise post completion of the CPT as compared to children in the distraction (Beta = -1.68, SE = 0.73, z = -2.30, p = .021, 95% CI [-3.10, -0.25]) and reappraisal (Beta = -1.24, SE = 0.72, z = -1.73, p = .084, 95% CI [-2.65, 0.17]) conditions. However, when self-reported pain and behavior were examined, no differences in pain severity (Wald Chi-squared (2) = 2.47, p = .292), or pain tolerance (Wald Chi-squared (2) = 1.38, p = 0.502) among the emotional regulation strategies were observed. Thus, significant findings were present for physiological markers of distress, but not for self-reported and behavioral measures. These findings suggest that in terms of physiological measures, specific emotional regulation strategies, such as distraction and reappraisal, may be more beneficial in reducing stress responses to painful medical procedures in pediatric patients with cancer as compared to reassurance. These results also demonstrate the importance of examining physiological outcomes in addition to self-report and behavioral outcomes

Low-Level Prenatal and Postnatal Blood Lead Exposure and Adrenocortical Responses to Acute Stress in Children

BACKGROUND: A few recent studies have demonstrated heightened hypothalamic–pituitary–adrenal (HPA) axis reactivity to acute stress in animals exposed to heavy metal contaminants, particularly lead. However, Pb-induced dysregulation of the HPA axis has not yet been studied in humans. OBJECTIVE: In this study, we examined children’s cortisol response to acute stress (the glucocorticoid product of HPA activation) in relation to low-level prenatal and postnatal Pb exposure. METHODS: Children’s prenatal blood Pb levels were determined from cord blood specimens, and postnatal lead levels were abstracted from pediatrician and state records. Children’s adrenocortical responses to an acute stressor were measured using assays of salivary cortisol before and after administration of a standard cold pressor task. RESULTS: Pb exposure was not associated with initial salivary cortisol levels. After an acute stressor, however, increasing prenatal and postnatal blood Pb levels were independently associated with significantly heightened salivary cortisol responses. CONCLUSIONS: Our results suggest that relatively low prenatal and postnatal blood lead levels— notably those below the 10 µg/dL blood lead level identified by the Centers for Disease Control and Prevention for public health purposes—can alter children’s adrenocortical responses to acute stress. The behavioral and health consequences of this Pb-induced HPA dysregulation in children have yet to be determined