Family Life Project Research Articles

Household Income Moderates Longitudinal Relations Between Neighborhood Child Opportunity Index and BMI Growth.

Background: To examine longitudinal associations of early neighborhood Child Opportunity Index 2.0 (COI) with children's BMI trajectories and identify whether household economic resources moderate relations of COI in infancy/toddlerhood and the preschool years to longitudinal BMI growth between 2 and 12 years. Methods: Family data (n = 1091) were drawn from the Family Life Project, a longitudinal study of families residing in rural high-poverty areas. Neighborhood COI was obtained for each developmental period: infancy/toddlerhood (2-15 months) and the preschool years (2-5 years). BMIs were created from anthropometrics collected at six time points. Results: Higher neighborhood COIs during the infancy/toddlerhood (β = -0.0130, p < 0.01) and preschool years (β = -0.0093, p < 0.05) were associated with lower BMI at 5 years of age; although the latter became nonsignificant after adjusting for infancy/toddlerhood COI. Both household income and time spent in poverty moderated associations of infancy/toddlerhood exposure to neighborhood COI with BMI change. Among children residing in not poor households, higher neighborhood level child opportunity was associated with a slower increase in BMI from 2 to 12 years (β = -0.0369, p < 0.05), and a lower BMI at 12 years (β = -0.0395, p < 0.05). Conclusions: Neighborhood COI during the infant and toddler years is longitudinally associated with child growth, and long-term associations are evident among children residing in not poor households. Future work is needed to better understand how family and neighborhood-level resources interact to influence obesity risk, particularly for those at high risk.

Considerations for Cell Type Heterogeneity in Pediatric Salivary DNA Methylation Analyses: Comparison of Reference Panels & Stratification by Estimated Cell Type Proportion.

Saliva is a widely used sample in epigenetic research with children due to its non-invasive nature. Since DNA methylation (DNAm) profile is cell type (CT) specific, salivary DNAm associations with exposures may be influenced by CT compositions, which is highly variable in saliva as it contains immune and buccal epithelial cells (BEC). Reference-based CT deconvolution and statistically adjusting estimated CT in DNAm analyses have become an increasingly common practice. However, careful examinations of how different reference panels may affect DNAm results and alternative approaches (e.g., stratification) are lacking. To scrutinize the best analytical strategies on pediatric salivary DNAm, the current study used 529 salivary DNAm samples obtained from children (mean age = 7.26 years, SD = 0.26 years) in the Family Life Project. Our results showed higher estimated CT variability with child than adult reference panels and highlighted the impact of these estimated CT discrepancies on DNAm associations with social variables (socioeconomic status). Stratifying salivary DNAm samples by BEC proportions detected a larger number of significant associations with biological variables (sex) and tissue-specific effect with cotinine level, a tobacco smoke-exposure biomarker. We provide analytical recommendations for future epigenetic research involving pediatric saliva samples.

Change in Child Opportunity Index in Early Childhood Is Associated with Youth BMI Growth.

Background: The neighborhood-level child opportunity index (COI) has been used in policy-based initiatives to identify and improve low-resource neighborhoods in order to impact child health. Understanding of how changes in COI can impact child growth, however, is lacking. Methods: Participants were 1124 children from the Family Life Project, a longitudinal birth cohort of families in rural, high-poverty areas. Youth anthropometrics were measured at eight assessments (ages 2 months through 12 years). Neighborhood COI was obtained at seven assessments (ages 2 months through 5 years) and used to create seven trajectory groups representing a change in COI: stayed low on all seven assessments, stayed moderate, stayed high, left low, declined from moderate, declined from high, and bounced around. Results: As hypothesized, moving from high COI neighborhoods into lower COI neighborhoods was associated with greater BMI growth and increased risk for obesity and severe obesity at 12 years. As hypothesized, the opposite effect, which approached significance at p = 0.056, was found among children who moved from low COI neighborhoods into higher COI neighborhoods. Specifically, moving into higher COI neighborhoods was associated with reduced BMI growth, and lower risk for severe obesity at 12 years. Conclusions: Moving into higher COI neighborhoods may be associated with healthier BMI growth, while the opposite effect may occur when moving into lower COI neighborhoods. Given the use of the COI in public health initiatives and growing evidence for its potential positive impact on child growth, future work is needed to replicate our findings among larger diverse samples.

Indirect effects of intrusive parenting on externalizing behaviors in socioeconomically disadvantaged children: A parallel mediation analysis

Although emerging research has identified intrusive parenting as a crucial risk factor for childhood externalizing behaviors, the mechanisms that underlie this relation warrant further investigation. Hence, the present study examined common executive functioning (EF) and expressive verbal abilities as parallel mediators in the associations between intrusive parenting and externalizing behaviors (i.e., hyperactivity/inattention, conduct problems) in preschool-aged children from low-income families. Data from the Family Life Project (N = 1050, Mage = 3 years 2 months) was analyzed using structural equation modelling. We found that common EF and expressive verbal abilities separately mediated the respective pathways from intrusive parenting to preschool-aged children’s hyperactivity/inattention problems and conduct problems. These findings held when key covariates-age, gender, household income, ethnicity, and state of residence—were controlled for. Notably, our findings provide evidence of potentially differing processes that explain the link between intrusive parenting and separable aspects of externalizing behaviors in young children from disadvantaged families.

Within-person bidirectional associations between maternal cortisol reactivity and harsh parenting across infancy and toddlerhood.

To understand transactional associations between mothers' biological stress responsivity and parenting behaviors, we examined bidirectional effects between maternal cortisol reactivity to observing their children during distress-eliciting paradigms and harsh parenting across infancy and toddlerhood using longitudinal data from the Family Life Project (N = 1,292, 41.5% African American). Children completed a series of distress-eliciting laboratory paradigms when they were 7, 15, and 24 months old, and mothers observed their children during the paradigms. Maternal cortisol reactivity was computed as a residualized change score from baseline to 20 min postparadigm, controlling for the time of day the saliva sample was collected. Harsh parenting was measured using five items from the Home Observation Measurement of the Environment inventory. A random-intercept cross-lagged panel model revealed that increases in maternal cortisol reactivity at 15 months predicted subsequent increases in harsh parenting at 24 months. Similarly, increases in harsh parenting at 15 months predicted increases in maternal cortisol reactivity at 24 months. Findings indicate that increased cortisol reactivity to children's distress in early toddlerhood may indicate a risk for harsh parenting in late toddlerhood and that increases in harsh parenting can also negatively impact mothers' stress physiology over time. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

An introduction to inverse probability weighting and marginal structural models: The case of environmental tobacco exposure and attention deficit/hyperactivity disorder behaviors.

Developmental scientists routinely examine how a focal predictor relates to some aspect of children's development. Although covariate adjustment is typically used to test hypotheses, propensity score-based methods, including inverse probability of treatment weighting (IPTW) and marginal structural models (MSM), can strengthen inference and answer more nuanced, developmentally relevant questions. This article provides a didactic introduction to IPTW and MSM methods and demonstrates their use for testing the impact of environmental smoke exposure (continuous treatment) from 6 to 90 months on parent-reported attention deficit hyperactivity disorder behaviors in first grade for 1,053 children (51% male, 44% Black) in the Family Life Project. We highlight differences that result both in conclusions and in the evaluation of assumptions for IPTW and MSM relative to more traditional covariate adjustment methods. Sample Stata syntax is provided. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

The salivary metabolome of children and parental caregivers in a large-scale family environment study

Human metabolism is complex, and is impacted by genetics, cohabitation, diet, health, and environmental inputs. As such, we applied untargeted LC-MS metabolomics to 1425 saliva samples from a diverse group of elementary school-aged children and their caregivers collected during the Family Life Project, of which 1344 were paired into caregiver/child dyads. We compared metabolomes within and between homes, performed population-wide “metabotype” analyses, and measured associations between metabolites and salivary biomeasures of inflammation, antioxidant potential, environmental tobacco smoke (ETS) exposure, metabolic regulation, and heavy metals. Children and caregivers had similar salivary metabolomes, and dyad explained most metabolomic variation. Our data clustered into two groups, indicating that “metabotypes” exist across large populations. Lastly, several metabolites—putative oxidative damage-associated or pathological markers—were correlated with the above-mentioned salivary biomeasures and heavy metals. Implications of the family environment’s effects on metabolomic variation at population, dyadic, and individual levels for human health are discussed.

Trajectories of maternal depressive symptoms from infancy through early childhood: The roles of perceived financial strain, social support, and intimate partner violence.

Although new mothers are at risk of heightened vulnerability for depressive symptoms, there is limited understanding regarding changes in maternal depressive symptoms over the course of the postpartum and early childhood of their child's life among rural, low-income mothers from diverse racial backgrounds. This study examined distinct trajectories of depressive symptoms among rural low-income mothers during the first five years of their child's life, at 6, 15, 24, and 58 months, using data from the Family Life Project (N = 1,292). Latent class growth analysis identified four distinct trajectories of maternal depressive symptoms, including Low-decreasing (50%; n = 622), Low-increasing (26%; n = 324), Moderate-decreasing (13%; n = 156), and Moderate-increasing (11%; n = 131) trajectories. Multinomial logistic regression demonstrated that higher perceived financial strain and intimate partner violence, and lower social support predicted higher-risk trajectories (Low-increasing, Moderate-decreasing, and Moderate-increasing) relative to the Low-decreasing trajectory. Compared to the Low-decreasing trajectory, lower neighborhood safety/quietness predicted to the Low-increasing trajectory. Moreover, lower social support predicted the Moderate-increasing trajectory, the highest-risk trajectory, compared to those in Moderate-decreasing. The current analyses underscore the heterogeneity on patterns of depressive symptoms among rural, low-income mothers, and that the role of both proximal and broader contexts contributing to distinct trajectories of maternal depressive symptoms over early childhood.

Associations among symbolic functioning, joint attention, expressive communication, and executive functioning of children in rural areas

The purpose of this study is to investigate the relations among children’s symbolic functioning at 15 months, joint attention at 24 months, expressive communication at 24 and 36 months, and executive functioning at 36 months. With the sample from rural areas in the United States collected by the Family Life Project (N = 1,008), a longitudinal data analysis was conducted. The results of structural equation modeling suggested that children’s symbolic functioning at 15 months and children’s executive functioning at 36 months was directly related to each other. These two variables were also indirectly related to each other through joint attention at 24 months and expressive communication at 24 and 36 months. Psychological distancing and verbal and nonverbal communication were used to explain the role symbolic functioning plays in the development of executive functioning during the second and the third years of children’s lives.

Increased risk of attention-deficit/hyperactivity disorder in adolescents with high salivary levels of copper, manganese, and zinc

Exposure to toxic heavy metals has been associated with the development of attention-deficit/hyperactivity disorder (ADHD). However, fewer studies have examined the associations between abnormal levels of essential trace metals and ADHD, and none have done so using saliva. We investigated whether salivary metals were associated with ADHD in adolescents aged 12 from the Family Life Project (FLP) using a nested case–control study design that included 110 adolescents who met diagnostic criteria for inattentive (ADHD-I), hyperactive-impulsive (ADHD-H), or combined type ADHD (ADHD-C) (cases) and 173 children who did not (controls). We used inductively coupled plasma optical emission spectrophotometry to measure chromium, copper, manganese, and zinc in saliva samples. We employed logistic regression models to examine associations between quartile levels of individual metals and ADHD outcomes by subtype. Salivary copper levels were significantly associated with increased odds of any ADHD diagnosis (OR = 3.31, 95% CI: 1.08–10.12; p = 0.04) and with increased odds of ADHD-C diagnosis (OR = 8.44, 95% CI: 1.58–45.12; p = 0.01). Salivary zinc levels were significantly associated with increased odds of ADHD-C diagnosis (OR = 4.06, 95% CI: 1.21–13.69; p = 0.02). Salivary manganese levels were also significantly associated with increased odds of ADHD-C diagnosis (OR = 5.43, 95% CI: 1.08–27.27, p = 0.04). This is the first study using saliva to assess metal exposure and provide a potential link between salivary levels of copper, manganese, and zinc and ADHD diagnoses in adolescents. Public health interventions focused on metal exposures might reduce ADHD incidence in low-income, minority communities.

Maternal Psychological Symptom Trajectories From 2 to 24 Months Postpartum: A Latent Class Growth Analysis.

Postpartum women experience an array of psychological symptoms that are associated with adverse health behaviors and outcomes including postpartum suicidal ideation and long-term depression. To provide early management of postpartum psychological symptoms, it is important to understand how the symptom experiences change over time. The aim of this study was to examine maternal psychological symptom trajectories over 2, 6, 15, and 24 months postpartum using latent class growth analysis and to examine how each trajectory is associated with maternal depression outcome at 24 months. We used secondary data from the Family Life Project ( N = 1,122) and performed multitrajectory latent class growth analysis based on four observed symptom variables (depression, anxiety, somatization, and hostility). After the final model was identified, bivariate analyses were conducted to examine the association between each trajectory and (a) individual characteristics and (b) outcome (Center for Epidemiologic Studies Depression Scale [CES-D]) variables. A four-class model was selected for the final model because it had better fit indices, entropy, and interpretability. The four symptom trajectories were (a) no symptoms over 24 months, (b) low symptoms over 24 months, (c) moderate symptoms increasing over 15 months, and (d) high symptoms increasing over 24 months. More postpartum women with the trajectory of high symptoms increasing over 24 months (Trajectory 4) were in low economic status (92.16%), unemployed (68.63%), or did not complete 4-year college education (98.04%). Most postpartum women (95.56%) in Trajectory 4 also had higher CES-D cutoff scores, indicating a possible clinical depression at 24 months postpartum. Postpartum women who experience increasing symptom trajectories over 15-24 months (Trajectories 3 and 4) could benefit from tailored symptom management interventions provided earlier in the postpartum period to prevent persistent and worsening symptom experiences.

Changes in maternal psychological symptom profiles from 2 to 6months postpartum: an application of latent transition analysis.

To identify subgroups of postpartum women with different psychological symptom profiles at 2 and 6months postpartum and to examine how they transition between symptom profiles over time using latent transition analysis (LTA). We used secondary data from the Family Life Project (N = 1,117) and performed LTA based on observed variables (depression, anxiety, somatization, and hostility). We examined transition probabilities and changes in latent status prevalence from 2 to 6months postpartum. Considering the known influences of social determinants of health on psychological symptoms, bivariate analyses were conducted to describe the characteristics of different transition patterns. A 3-class model with better fit indices, entropy, and interpretability was selected. Based on symptom severity, the identified profiles were Profile 1: Low, Profile 2: Moderate, and Profile 3: High. From 2 to 6months postpartum, the prevalence of low symptom profile decreased (82 to 78.2%) while the prevalence increased for moderate (15.8 to 17.5%) and high symptom profiles (2.2 to 4.4%). For all profiles, it was most likely for postpartum women to stay in the same profile from 2 to 6months (low to low, moderate to moderate, and high to high). Those in persistent or worsening transition patterns were significantly younger or had less social support or education. Postpartum women in moderate or high symptom profiles at 2months were most likely to stay in the same profile at 6months postpartum, indicating persistent symptom burden. Clinicians should consider providing early, targeted support to prevent persistent symptom burden.

Salivary uric acid across child development and associations with weight, height, and body mass index.

Obesity during childhood is a serious and growing chronic disease with consequences for lifelong health. In an effort to advance research into the preclinical indicators of pediatric obesity, we examined longitudinal assessments of uric acid concentrations in saliva among a cohort of healthy children from age 6-months to 12-years (n's per assessment range from 294 to 727). Using data from a subsample of participants from the Family Life Project (an Environmental influences on Child Health Outcomes Program cohort), we: (1) characterized salivary uric acid (sUA) concentrations from infancy to early adolescence by sex and race; (2) assessed changes in sUA levels across development; and (3) evaluated associations between sUA concentrations and measures of child weight, height, and body mass index (BMI). Across four assessments conducted at 6-, 24-, 90-, and 154-months of age, 2,000 saliva samples were assayed for UA from 781 participants (217 participants had sUA data at all assessments). There were no significant differences in sUA concentrations by sex at any assessment, and differences in sUA concentrations between White and non-White children varied by age. At the 90- and 154-month assessments, sUA concentrations were positively correlated with measures of child weight, height, and BMI (90-month: weight- ρ(610) = 0.13, p < 0.01; height- ρ(607) = 0.10, p < 0.05; BMI- ρ(604) = 0.13, p < 0.01; 154-month: weight- ρ(723) = 0.18, p < 0.0001; height- ρ(721) = 0.10, p < 0.01; BMI- ρ(721) = 0.17, p < 0.0001). Group based trajectory modeling identified two groups of children in our sample with distinct patterns of sUA developmental change. The majority (72%) of participants showed no significant changes in sUA across time ("Stable" group), while 28% showed increases in sUA across childhood with steep increases from the 90- to 154-month assessments ("Increasing" group). Children in the Increasing group exhibited higher sUA concentrations at all assessments (6-month: t(215) = -5.71, p < 0.001; 24-month: t(215) = -2.89, p < 0.01; 90-month: t(215) = -3.89, p < 0.001; 154-month: t(215) = -19.28, p < 0.001) and higher weight at the 24- and 90-month assessments (24-month: t(214) = -2.37, p < 0.05; 90-month: t(214) = -2.73, p < 0.01). Our findings support the potential utility of sUA as a novel, minimally-invasive biomarker that may help advance understanding of the mechanisms underlying obesity as well as further surveillance and monitoring efforts for pediatric obesity on a large-scale.

Association between Teacher Depressive Symptoms and Rural Children's Cognitive Outcomes: Indirect Relations with Early Care and Education Quality.

A subset of extant data (N=423) from the Family Life Project, a population-based study of Black and White families with low incomes from rural communities, were used to test for associations between teacher depressive symptoms and children's cognitive outcomes at 36 months. A second aim tested whether early care and education (ECE) quality mediated such relations. Results indicated that although the associations were in the expected negative direction, teacher depressive symptoms were not directly significantly related to any child cognitive outcome. However, relations were completely mediated by indirect pathways with ECE quality, meaning that teacher depressive symptoms affected child cognitive outcomes entirely via decreases in ECE quality (ES= -0.04), particularly for receptive vocabulary. Implications for teacher mental health and psychological wellbeing as well as ECE quality in rural communities are discussed.

Profiles of perceived resources among low-income, rural mothers: Prospective associations with maternal and child outcomes.

Low-income, rural families face significant mental health risks. However, the understanding of resources associated with mental health risks is limited. The present study investigated the associations between perceived resources of low-income, rural mothers, and longitudinal maternal and child outcomes. This study utilized longitudinal data from the Family Life Project (N=1203), from US rural areas with high poverty rates. Mothers reported their resources at 6-month postpartum, and their levels of depression, anxiety, and role overload were assessed at 2-year postpartum. Mothers reported their children's behavioral problems at 3 years old. Using a person-centered approach, we identified four maternal profiles: lower resources (7.1%); higher intra-family support (11.1%); higher inter-family support (20.8%); and higher resources (60.9%). In general, the higher resource profile was associated with lower mental health concerns of mothers and lower levels of behavior problems of children. Mothers in the higher intra-family support profile had disproportionately higher role overload. Children of mothers in the higher inter-family support profile showed disproportionately higher behavioral problems. Maternal partner status and education were significant predictors of resource profiles. Findings support the heterogeneity in perceived resources among low-income, rural families and different risk levels. Identifying these subgroups has significant implications for policy and interventions aimed toward this vulnerable population.

Measuring adolescents’ eating in the absence of hunger in the home environment using subjective and objective measures

Eating in the absence of hunger (EAH) has been identified as a behavioral phenotype for obesity. Few studies have reported on objective measures of EAH in adolescents, and fewer yet have objectively measured EAH in a naturalistic, home setting. The purpose of this paper was to examine relations between objective, adolescent-report and parent-report measures of EAH, and to examine variation by sex and race. Participants included 295 predominantly low-income and rural adolescents (mean age = 14.2 ± 0.6 years) and their parents, drawn from the Family Life Project. An EAH task was administered in the home following an ad-libitum meal and compulsory milkshake; EAH was also reported on a web-based survey (both adolescent and parent reports) and adolescents’ BMIz was calculated from height and weight, measured in the home or self-reported on the web survey. A high degree of variability in EAH intake was observed (range = 8–741 kcals). Parent and adolescent reports of EAH were weakly correlated and unrelated to observed EAH consumption; only adolescent reports of EAH were related to their BMIz. Several relations varied by sex and race. Positive associations between reported and observed EAH was only observed in girls, and positive associations between observed EAH and BMI was only observed in boys and in white adolescents. Overall EAH consumption was significantly greater in boys and in white adolescents. These findings suggest that EAH can be measured in adolescents in the home. In this sample of youth experiencing rural poverty, this home-based measure appears most valid for white adolescent girls.

Head Start in Low-Wealth, Rural Communities: Evidence from the Family Life Project

ABSTRACT This study utilized data from the Family Life Project (FLP) to examine Head Start children’s school readiness skills at the end of preschool in comparison to two other care groups: home-based care and other center-based care. The FLP study enrolled a birth-cohort of 1,292 children born in two historically low-wealth, rural regions of the U.S., with oversampling of children from low-wealth families and African American families. Propensity score weighting adjusted for differences in baseline child and family characteristics between the Head Start participants and children in the two alternative care groups. Research Findings: The results indicated that the Head Start group had higher literacy skills at the end of preschool in comparison to the home-based care group (β = 0.36), but no other reliable differences emerged between these groups in school readiness skills. No reliable differences emerged between the Head Start and other center-based care groups in school readiness skills. Practice and Policy: Implications for educational practice and policy were discussed in relation to Head Start programming in rural America, including potential targets for program improvement and strategies for program expansion.

Sensitive caregiving and reward responsivity: A novel mechanism linking parenting and executive functions development in early childhood.

Sensitive caregiving is an essential aspect of positive parenting that influences executive functions development, but the mechanisms underlying this association are less clear. Using data from the Family Life Project, a large prospective longitudinal sample of 1292 families residing in rural, predominately low-income communities, the current study examined whether sensitive caregiving impacts executive functions development by shaping behavioral reward processing systems in early postnatal life. Results indicated that higher levels of sensitive caregiving during infancy were associated with heightened reward responsivity at age 4, which in turn predicted superior executive functions ability at age 5. Notably, children's reward responsivity partially mediated the relationship between sensitive caregiving in infancy and executive functions ability at school entry. These findings add to prior work on early experience and children's executive functions and highlight caregiver scaffolding of developing reward processing systems as a potential foundational mechanism for supporting adaptive behavior and self-regulation across the lifespan.

A dynamic systems perspective towards executive function development: Susceptibility at both ends for inhibitory control.

In light of the dynamic systems perspective, the current study expanded existing literature by examining the moderating effect of maternal sensitivity on the quadratic association between infant negative reactivity and future executive function development. Using a longitudinal, multimethod design, we addressed executive function development among preschoolers. This study utilized data from the Family Life Project (N = 1292). Infant negative reactivity at 6 months, maternal sensitivity across first 3 years, and executive functions during preschool age were observational assessed. A path model with moderation analyses revealed a U-shaped quadratic association between infant negative reactivity and preschoolers' inhibitory control, only when maternal sensitivity was high. The results suggest that maternal sensitivity may assist infants with both low and high, but not moderate, levels of negative reactivity towards better executive function development. Findings support the ongoing nonlinear person-environment interplay during early years of life.

Profiles of family-based social experiences in the first 3 years predict early cognitive, behavioral, and socioemotional competencies.

This study utilized latent profile analyses to identify unique configurations of children's family-based social experiences during the first 3 years of life and examine differences across profiles with respect to developmental outcomes at 36 and 48 months of age. Seven family process variables were used: maternal emotional functioning, maternal sensitivity, negative controlling parenting, cognitive stimulation, corporal punishment, adult-adult aggression, and household disorganization. Data were collected by the Family Life Project (N = 1,087), a longitudinal study of families living in low-wealth, nonurban areas, for whom the biological mother was the child's primary caregiver from 6 to 36 months of age. On average, mothers were 26 years of age at 2 months of child age. Approximately 36% of the families lived below the federal poverty limit, and 41% identified as Black; 49% of children were female. Latent profile analyses identified four groups: (a) positive exposure (b) average exposure, (c) problematic adult functioning, and (d) problematic parenting . Comparisons indicate that children in the positive exposure profile had the highest levels of socioemotional and cognitive outcomes compared to overall profiles. Children in the problematic adult functioning and problematic parenting profiles had the most problematic child outcomes, with children in the problematic parenting profile scoring lowest overall. Results indicate that there is configural heterogeneity in family-based social experiences at the highest levels of risk and that exposures to problematic parenting may be more consequential for later child outcomes than exposures to problematic adult functioning in the absence of compromised caregiving. (PsycInfo Database Record (c) 2022 APA, all rights reserved).