Effects Of Prenatal Cocaine Exposure Research Articles

Prenatal cocaine exposure and its influence on pediatric epigenetic clocks and epigenetic scores in humans

The investigation of the effects of prenatal cocaine exposure (PCE) on offspring has been inconsistent, with few studies investigating biological outcomes in humans. We profiled genome-wide DNA methylation (DNAm) of umbilical cord blood (UCB) from newborns with (n = 35) and without (n = 47) PCE. We used DNAm data to (1) assess pediatric epigenetic clocks at birth and (2) to estimate epigenetic scores (ES) for lifetime disorders. We generated gestational epigenetic age estimates (DNAmGA) based on Knight and Bohlin epigenetic clocks. We also investigated the association between DNAmGA and UCB serum brain-derived neurotrophic factor (BDNF) levels. Considering the large-scale DNAm data availability and existing evidence regarding PCE as a risk for health problems later in life, we generated ES for tobacco smoking, psychosis, autism, diabetes, and obesity. A gene ontology (GO) analysis on the CpGs included in the ES with group differences was performed. PCE was associated with lower DNAmGA in newborns, and this effect remained significant when controlling for potential confounders, such as blood cell type composition predicted by DNAm and obstetric data. DNAmGA was negatively correlated with BDNF levels in the serum of UCB. Higher tobacco smoking, psychosis, and diabetes ES were found in the PCE group. The GO analysis revealed GABAergic synapses as a potential pathway altered by PCE. Our findings of decelerated DNAmGA and ES for adverse phenotypes associated with PCE, suggest that the effects of gestational cocaine exposure on the epigenetic landscape of human newborns are detectable at birth.

Self-reported mental health outcomes in prenatally cocaine exposed adolescents at 17 years of age

ObjectiveAdverse developmental effects of prenatal cocaine exposure (PCE) are hypothesized to extend into late adolescence, yet few studies have investigated the association between PCE and late adolescent mental health outcomes. We examined the associations between PCE and self-reported mental health symptoms at age 17, controlling for biologic and environmental confounders. We further explored the potential moderating role of sex and the mediating role of earlier drug use by age 15 in the associations. Method327 (162 PCE; 165 non-cocaine exposed, NCE) urban adolescents, primarily African Americans, of low socioeconomic status, were prospectively recruited at birth for a longitudinal study and participated in the current study. We administered the Computerized Diagnostic Interview Schedule for Children-IV to assess their mental health symptoms at age 17. Alcohol, tobacco, and marijuana use by age 15 were assessed using biologic samples and self-reports. Confounders included other prenatal drug exposures, caregiving environment, and childhood maltreatment. ResultsAlthough no overall associations between PCE and mental health outcomes were observed, multivariate logistic regression models indicate girls with PCE were 3.60 times (95% CI = 1.45–8.96, p = .006) more likely to have symptoms of oppositional defiance disorder than girls with NCE. This relationship was partially mediated by marijuana use by age 15. ConclusionContinued studies into emerging adulthood will further elucidate the long-term mental health outcomes associated with PCE.

Sex moderated and RSA mediated effects of prenatal cocaine exposure on behavior problems at age 7

This study was designed to assess whether prenatal cocaine exposure (PCE) is associated with sex differences in behavior problems in middle childhood and whether there are sex differences in the way in which parasympathetic functioning mediates the relations between PCE and behavior problems within a diverse low-income sample. Participants included 164 high risk mother-child dyads including 89 PC exposed children and 75 control children participating in an ongoing longitudinal study. Respiratory sinus arrhythmia (RSA) was measured to assess parasympathetic functioning at 13 months of age and maternal reports of child behavior problems were collected at 7 years of age. Results revealed no significant association between PCE and behavior problems for the full sample. A 2 × 2 Anova revealed a significant interaction between PCE and child sex on internalizing, externalizing, and total behavior problems (F (3, 160) = 13.45, p < .001) with cocaine exposed females averaging the highest behavior problem scores. Results also revealed a statistically significant indirect effect linking cocaine exposure to lower externalizing problems via lower baseline RSA among males. Findings indicate that cocaine exposed females may be more vulnerable to developing behavior problems than cocaine exposed males and that high baseline RSA may present a sex specific risk factor for externalizing problems among exposed males.

Preschool blood lead levels, language competency, and substance use in adolescence

BackgroundElevated lead levels in children are a persistent public health problem, particularly in urban areas in the United States, yet few prospective studies have examined the association of childhood lead levels with substance use in adolescence. ObjectivesTo determine the association of early lead levels with adolescent substance use and whether childhood IQ, language skills, and externalizing (aggressive and disruptive) behavior mediate the association, controlling for confounding biological and environmental factors. MethodsThe participants (N = 265) were a subsample of a prospective birth cohort study on the developmental effects of prenatal cocaine exposure in the Midwest United States. Blood lead levels (BLL) were assessed at age 4, IQ at age 11, language skills and externalizing behavior at age 12, and substance (alcohol, tobacco, marijuana) use and substance use-related problems at age 15. Biologic assays (hair, urine, bloodspots), along with self-report, were utilized to determine adolescent substance use. Path analyses were conducted to examine the direct and indirect associations of BLL with adolescent substance use. ResultsThe children's mean BLL at 4 years of age was 7.07 (SD = 4.12) μg/dL. Approximately 31% of adolescents used tobacco or marijuana, 40% used alcohol, and 23% reported experiencing substance use-related problems at age 15.7 (SD = 0.28). Elevated BLL was related to a higher likelihood of substance use. Childhood language skills fully mediated the relationship of BLL with substance use-related problems. IQ was related to neither substance use nor substance use-related problems. DiscussionElevated BLL in preschool years is a risk factor for adolescent substance use and related problems. Early screening and intervention for language impairment may reduce substance use-related problems.

Profiles of individual assets and mental health symptoms in at-risk early adolescents

IntroductionFew studies investigated the combined patterns of individual assets (e.g., social competence, positive identity) and mental health symptoms (MHS) in adolescents. This study examined the patterns of early adolescents' individual assets and MHS and whether identified patterns were associated with later adolescents’ outcomes. MethodsParticipants were 352 (164 boys, 188 girls) adolescents who were primarily African-American and from low socioeconomic status families, participating in a prospective study of the effects of prenatal cocaine exposure from birth in the Midwest United States. Individual assets, using the Developmental Assets Profile, and MHS, using the Youth Self-Report, were assessed at age 12. Substance use, via self-report and biologic assays, early (before age 15) sexual behaviors, and behavioral adjustment were assessed at age 15. ResultsLatent profile analysis indicated four distinctive profiles: low assets with elevated MHS (P1, n = 54, 15.3%); adequate assets with thought and social problems (P2, n = 84, 23.9%); low assets without MHS (P3, n = 101, 28.7%); and high assets without MHS (P4, n = 113, 32.1%). Children in the profile with high assets without MHS (P4) were more likely to have a higher IQ and to be in a more optimal environment (higher parental monitoring and less family conflict) than those in other profiles. Although profiles with MHS were associated with adolescent risk behaviors, this relationship was more pronounced for girls than for boys. ConclusionsGirls in the low assets with elevated MHS (P1) should be a primary concern for preventive intervention. Our study demonstrates the heterogeneity of individual patterns of adaptation and maladaptation.

Individual assets and problem behaviors in at-risk adolescents: A longitudinal cross-lagged analysis

Autoregressive cross-lagged structural equation modeling was conducted to examine longitudinal relationships between individual assets (social competence, positive values and identity) and problem behaviors in 373 adolescents (174 boys, 199 girls) who participated in a prospective study on the effects of prenatal cocaine exposure from birth. More behavioral problems at age 12 were related to fewer individual assets at age 15, while greater individual assets were related to more behavioral problems, with a non-significant yet nuanced (p = .076) gender difference. More problem behaviors were associated with decreased individual assets in girls, yet greater individual assets were associated with more problem behaviors in boys. Efforts to promote individual assets may not lower the risk of engaging in problem behaviors especially among boys. Continued studies into adulthood will uncover how individual assets and problem behaviors in childhood and adolescence may affect social and vocational adjustment in this high risk population.

Prenatal Cocaine Exposure Alters BDNF-TrkB Signaling in the Embryonic and Adult Brain

Prenatal cocaine exposure remains a major public health concern because of its adverse effects on cognitive function. Although the molecular mechanisms underlying the cognitive impairment are not fully understood, brain-derived neurotrophic factor (BDNF) signaling via its receptor tyrosine kinase B (TrkB) is emerging as a potential candidate. We used a mouse model to examine the impact of ongoing cocaine exposure on BDNF expression in the dorsal forebrain on embryonic day 15 (E15) as well as the long-term effects of prenatal cocaine exposure on BDNF-TrkB signaling in the frontal cortex in early postnatal (postnatal day 16; P16) and adult (P60) male and female mice. We found that ongoing cocaine exposure decreased BDNF expression in the E15 dorsal forebrain, prenatal cocaine exposure did not alter BDNF or TrkB (total or phosphorylated) expression in the frontal cortex at P16, and that the prenatal cocaine exposure produced significant increases in BDNF, the activated (phosphorylated) form of TrkB, as well as Bdnf mRNA in the frontal cortex at P60. The increase in BDNF protein and mRNA expression at P60 was concurrent with hyperacetylation of histone H3 at the Bdnf promoter in the frontal cortex. The increase in frontal cortical BDNF and activated TrkB at P60 occurred in male but not female mice. Thus, our data demonstrate that ongoing cocaine exposure produces a decrease in BDNF expression in the embryonic brain, and that prenatal cocaine exposure produces a sex-specific increase in frontal cortical BDNF-TrkB signaling at P60 only in male mice. Lastly, hyperacetylation of histone H3 at the Bdnf promoter is one epigenetic mechanism mediating the effects of prenatal cocaine exposure on Bdnf expression at P60 in male mice.

CYP3A5 Mediates Effects of Cocaine on Human Neocorticogenesis: Studies using an In Vitro 3D Self-Organized hPSC Model with a Single Cortex-Like Unit.

Because of unavoidable confounding variables in the direct study of human subjects, it has been difficult to unravel the effects of prenatal cocaine exposure on the human fetal brain, as well as the cellular and biochemical mechanisms involved. Here, we propose a novel approach using a human pluripotent stem cell (hPSC)-based 3D neocortical organoid model. This model retains essential features of human neocortical development by encompassing a single self-organized neocortical structure, without including an animal-derived gelatinous matrix. We reported previously that prenatal cocaine exposure to rats during the most active period of neural progenitor proliferation induces cytoarchitectural changes in the embryonic neocortex. We also identified a role of CYP450 and consequent oxidative ER stress signaling in these effects. However, because of differences between humans and rodents in neocorticogenesis and brain CYP metabolism, translation of the research findings from the rodent model to human brain development is uncertain. Using hPSC 3D neocortical organoids, we demonstrate that the effects of cocaine are mediated through CYP3A5-induced generation of reactive oxygen species, inhibition of neocortical progenitor cell proliferation, induction of premature neuronal differentiation, and interruption of neural tissue development. Furthermore, knockdown of CYP3A5 reversed these cocaine-induced pathological phenotypes, suggesting CYP3A5 as a therapeutic target to mitigate the deleterious neurodevelopmental effects of prenatal cocaine exposure in humans. Moreover, 3D organoid methodology provides an innovative platform for identifying adverse effects of abused psychostimulants and pharmaceutical agents, and can be adapted for use in neurodevelopmental disorders with genetic etiologies.

Pathways to adolescent sexual risk behaviors: Effects of prenatal cocaine exposure

BackgroundTo assess the impact of prenatal cocaine exposure (PCE) on adolescent sexual risk behaviors. Externalizing behavior, teen substance use, and early sexual intercourse were examined as pathways mediating the effects of PCE on sexual risk behaviors. MethodsAdolescents (N=364; 185 PCE, 179 non-cocaine exposure (NCE); 205 girls, 159 boys), primarily African–American and of low socioeconomic status, were prospectively enrolled in a longitudinal study at birth. Risky sexual behaviors were assessed at ages 15 and 17. Externalizing behavior at 12 years was assessed with the Youth Self-Report. Substance use, via self-report and biologic assays, and early (before age 15) sexual intercourse were assessed at age 15. Path analyses with the weighted least squares estimator with mean and variance adjustments were performed. ResultsThe final structural equation model-based path model, χ2=31.97 (df=27), p=.23, CFI=.99, TLI=.99, RMSEA=.021, WRMR=.695, indicated a direct effect of PCE on sexual risk behavior (β=.16, p=.02). Although PCE was related to greater externalizing behavior (β=.14, p=.009), which in turn, predicted early sexual intercourse (β=.16, p=.03), leading to sexual risk behavior (β=.44, p<.001), bootstrapping indicated a non-significant indirect effect (β=.01, p>.10). Substance use was correlated with early sexual intercourse (r=.60, p<.001) and predicted sexual risk behavior by age 17 (β=.31, p=.01). ConclusionsPrenatal cocaine exposure was related to more engagement in sexual risk behaviors, suggesting the importance of reducing substance use among pregnant women as a means of prevention of offspring substance use and sexual risk behavior.

Prenatal Cocaine Exposure and Cardiometabolic Disease Risk Factors in 18 to 20 Year Old African Americans

The long-term effects of prenatal cocaine exposure (PCE) on physical health are largely unknown. No human studies support or refute a relationship between PCE and the long-term risk for cardiovascular and/or metabolic disease. We investigated the association of PCE on primary cardiometabolic disease risk factors in African Americans (AA) aged 18 to 20 years. Cohort, longitudinal, prospective. Miami-Dade County, Florida, and the University of Miami Miller School of Medicine/Jackson Memorial Medical Center. Healthy full-term inner-city AA adolescents (aged 18 to 20 years, n=350) previously enrolled at birth from 1990-1993. Fasting serum insulin, glucose, lipids, and high-sensitivity C-reactive protein; systolic and diastolic blood pressures; and the components and prevalence of the metabolic syndrome. There were no PCE-associated differences in cardiometabolic disease risk factors including the metabolic syndrome and its individual components in AAs aged 18 to 20 years. The results of our study do not support an association between PCE and increased cardiometabolic disease risk in AAs aged 18 to 20 years. Whether PCE is associated with cardiovascular or metabolic disease in adulthood would require further investigation.

Effects of prenatal cocaine exposure on early sexual behavior: Gender difference in externalizing behavior as a mediator

BackgroundPrenatal cocaine exposure (PCE) is associated with increased risk for externalizing behavior problems; childhood externalizing behavior problems are linked with subsequent early sexual behavior. The present study examined the effects of PCE on early sexual initiation (sexual intercourse prior to age 15) and whether externalizing behavior in preadolescence mediated the relationship. MethodsThree hundred fifty-four (180 PCE and 174 non-cocaine exposed; 192 girls, 142 boys), primarily African-American, low socioeconomic status, 15-year-old adolescents participated in a prospective longitudinal study. Adolescents’ sexual behavior was assessed at 15 years using the Youth Risk Behavior Surveillance System. Externalizing behavior was assessed at 12 years using the Youth Self-Report. ResultsLogistic regression models indicated that adolescents with PCE (n=69, 38%) were 2.2 times more likely (95% CI=1.2–4.1, p<.01) to engage in early sexual intercourse than non-exposed peers (n=49, 28%) controlling for covariates. This relationship was fully mediated by self-reported externalizing behavior in girls but not in boys, suggesting childhood externalizing behavior as a gender moderated mediator. Blood lead level during preschool years was also related to a greater likelihood of early sexual intercourse (OR=2.6, 95% CI=1.4–4.7, p<.002). Greater parental monitoring decreased the likelihood of early sexual intercourse, while violence exposure increased the risk. ConclusionsPCE is related to early sexual intercourse, and externalizing behavior problems mediate PCE effects in female adolescents. Interventions targeting externalizing behavior may reduce early sexual initiation and thereby reduce HIV risk behaviors and early, unplanned pregnancy in girls with PCE.

Effects of prenatal cocaine exposure on early sexual activity: Gender difference in externalizing behavior as a mediator

Effects of prenatal cocaine exposure on early sexual activity: Gender difference in externalizing behavior as a mediator

Effects of prenatal cocaine exposure and externalizing behavior on adolescent substance use (15–17 years)

Effects of prenatal cocaine exposure and externalizing behavior on adolescent substance use (15–17 years)

Effects of prenatal cocaine exposure on adolescent development

The associations between prenatal cocaine exposure (PCE) and adolescent behavior, cognitive development, and physical growth were examined in 219 15-year-olds who have participated in a longitudinal study since their fourth gestational month. During the first trimester, 42% of the women used cocaine, with use declining across pregnancy. At the 15-year follow-up, the caregivers were, on average, 43years old, had 13years of education, and 50% were African American. First trimester PCE was not associated with global cognitive development or with measures of learning and memory. First trimester PCE was significantly related to adolescent-reported delinquent behavior, poorer problem solving and abstract reasoning, and reduced weight, height, and head circumference at 15years. These results were significant after other factors that affect these domains were controlled in regression analyses. In addition, exposure to violence partially mediated the effect of PCE on delinquent behavior. These adolescent domains are important because they are predictors of poorer adult functioning.

Executive function and cortical thickness in youths prenatally exposed to cocaine, alcohol and tobacco

Small and detrimental, albeit inconsistent, effects of prenatal cocaine exposure (PCE) during early childhood have been reported. The teratogenic effects of prenatal alcohol (PAE) and tobacco exposure (PTE) on neurobehavior are more firmly established than PCE. We tested if co-exposure to all three drugs could be related to greater differences in brain structure than exposure to cocaine alone. Participants (n=42, PCE=27; age range=14–16 years) received an executive function battery prior to a T1-weighted 3T structural MRI scan. Cortical thickness was measured using FreeSurfer (v5.1). Fetal drug exposure was quantified through maternal self-reports usage during pregnancy. Using general linear modeling, we found no main effects of PCE on cortical thickness, but significant main effects of PAE and PTE in superior and medial frontal regions, after co-varying for the effects of age, sex, and each drug of exposure. Significant alcohol-by-tobacco interactions, and significant cocaine-by-alcohol interactions on cortical thickness in medial parietal and temporal regions were also observed. Poly-drug exposure and cognitive function also showed significant interactions with cortical thickness: lower cortical thickness was associated with better performance in PCE-exposed adolescents. Results suggest that although children with PCE have subtle but persistent brain cortical differences until mid-to-late adolescence.

Effects of prenatal cocaine exposure on pubertal development

The purpose of the current study was to examine the relationship between prenatal cocaine exposure (PCE) and pubertal development. Children (n=192; 41% with PCE) completed the Pubertal Development Scale (Petersen et al. 1988) and provided salivary dehydroepiandrosterone (DHEA) samples at 6month intervals from 11 to 13years. PCE was examined as a predictor of pubertal status, pubertal tempo, and DHEA levels in mixed models analyses controlling for age, sex, environmental risk, neonatal medical problems, other prenatal exposures, and BMI. PCE interacted with age such that PCE predicted slower pubertal tempo during early adolescence. PCE also interacted with age to predict slower increases in DHEA levels during early adolescence. These findings suggest that PCE may affect pubertal development and, if slower pubertal tempo continues, could lead to delayed pubertal status in mid-adolescence.

Prenatal cocaine exposure and adolescent neural responses to appetitive and stressful stimuli.

Preclinical research has demonstrated the effects of prenatal cocaine exposure (PCE) on brain regions involved in emotional regulation, motivational control, and addiction vulnerability-eg, the ventral striatum (VS), anterior cingulate (ACC), and prefrontal cortex (PFC). However, little is known about the function of these regions in human adolescents with PCE. Twenty-two adolescents with PCE and 22 age-, gender-, and IQ-matched non-cocaine exposed (NCE) adolescents underwent functional magnetic resonance imaging (fMRI) during exposure to individually personalized neutral/relaxing, stressful, and favorite-food cues. fMRI data were compared using group-level two-tailed t-tests in the BioImage Suite. In comparison with NCE adolescents, PCE adolescents had reduced activity within cortical and subcortical brain regions, including the VS, ACC, and medial and dorslolateral PFC during exposure to favorite-food cues but did not differ in neural responses to stress cues. Subjective food craving was inversely related to dorsolateral PFC activation among PCE adolescents. Among PCE adolescents, subjective anxiety ratings correlated inversely with activations in the orbitofrontal cortex and brainstem during the stress condition and with ACC, dorsolateral PFC, and hippocampus activity during the neutral-relaxing condition. Thus adolescents with PCE display hypoactivation of brain regions involved in appetitive processing, with subjective intensities of craving and anxiety correlating inversely with extent of activation. These findings suggest possible mechanisms by which PCE might predispose to the development of addictions and related disorders, eg, substance-use disorders and binge-eating.

Effects of Prenatal Cocaine Exposure on Social Development in Mice

Prenatal cocaine exposure (PCE) in humans and animals has been shown to impair social development. Molecules that mediate synaptic plasticity and learning in the medial prefrontal cortex (mPFC), specifically brain-derived neurotrophic factor (BDNF) and its downstream signaling molecule, early growth response protein 1 (egr1), have been shown to affect the regulation of social interactions (SI). In this study we determined the effects of PCE on SI and the corresponding ultrasonic vocalizations (USVs) in developing mice. Furthermore, we studied the PCE-induced changes in the constitutive expression of BDNF, egr1 and their transcriptional regulators in the mPFC as a possible molecular mechanism mediating the altered SI. In prenatal cocaine-exposed (PCOC) mice we identified increased SI and USV production at postnatal day (PD) 25, and increased SI but not USVs at PD35. By PD45 the expression of both social behaviors normalized in PCOC mice. At the molecular level, we found increased BDNF exon IV and egr1 mRNA in the mPFC of PCOC mice at PD30 that normalized by PD45. This was concurrent with increased EGR1 protein in the mPFC of PCOC mice at PD30, suggesting a role of egr1 in the enhanced SI observed in juvenile PCOC mice. Additionally, by measuring the association of acetylation of histone 3 at lysine residues 9 and 14 (acH3K9,14) and MeCP2 at the promoters of BDNF exons I and IV and egr1, our results provide evidence of promoter-specific alterations in the mPFC of PCOC juvenile mice, with increased association of acH3K9,14 only at the BDNF exon IV promoter. These results identify a potential PCE-induced molecular alteration as the underlying neurobiological mechanism mediating the altered social development in juvenile mice.

Heart Rate Variability and Cocaine: a Systematic Review of Human Studies

Context:: Cocaine affects the autonomic nervous system (ANS). Cardiovascular effects of cocaine have been extensively studied by measures of mean heart rate (HR) or blood pressure (BP). The present systematic review aims to summarize the current evidence on the effects of cocaine on heart rate variability (HRV) in human subjects. Evidence Acquisition:: A systematic search of electronic databases (PUBMED, PSYNDEX and EMBASE) based on the “Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA)” statement was employed. Studies were screened for eligibility based on pre-specified inclusion criteria. The search revealed a total of 48 papers after removing duplicates. Eleven studies fitting the inclusion criteria were included in the systematic review. Results: Evidence on the effects of cocaine on HRV is rare. Existing studies either address the impact of acute cocaine administration in adults or the effect of prenatal cocaine exposure on infants in various age groups. Conclusions:: However, HRV is a promising index to study ANS function within this particular field of research. Several suggestions for future research are provided.

Prenatal Cocaine Exposure Uncouples mGluR1 from Homer1 and Gq Proteins

Cocaine exposure during gestation causes protracted neurobehavioral changes consistent with a compromised glutamatergic system. Although cocaine profoundly disrupts glutamatergic neurotransmission and in utero cocaine exposure negatively affects metabotropic glutamate receptor-type 1 (mGluR1) activity, the effect of prenatal cocaine exposure on mGluR1 signaling and the underlying mechanism responsible for the prenatal cocaine effect remain elusive. Using brains of the 21-day-old (P21) prenatal cocaine-exposed rats, we show that prenatal cocaine exposure uncouples mGluR1s from their associated synaptic anchoring protein, Homer1 and signal transducer, Gq/11 proteins leading to markedly reduced mGluR1-mediated phosphoinositide hydrolysis in frontal cortex (FCX) and hippocampus. This prenatal cocaine-induced effect is the result of a sustained protein kinase C (PKC)-mediated phosphorylation of mGluR1 on the serine residues. In support, phosphatase treatment of prenatal cocaine-exposed tissues restores whereas PKC-mediated phosphorylation of saline-treated synaptic membrane attenuates mGluR1 coupling to both Gq/11 and Homer1. Expression of mGluR1, Homer1 or Gα proteins was not altered by prenatal cocaine exposure. Collectively, these data indicate that prenatal cocaine exposure triggers PKC-mediated hyper-phosphorylation of the mGluR1 leading to uncoupling of mGluR1 from its signaling components. Hence, blockade of excessive PKC activation may alleviate abnormalities in mGluR1 signaling and restores mGluR1-regulated brain functions in prenatal cocaine-exposed brains.