Advanced glycation end product-modified low-density lipoprotein promotes pro-osteogenic reprogramming via RAGE/NF-κB pathway and exaggerates aortic valve calcification in hamsters.

BACKGROUND: Advanced glycation end product-modified low-density lipoprotein (AGE-LDL) is related to inflammation and the development of atherosclerosis. Additionally, it has been demonstrated that receptor for advanced glycation end products (RAGE) has a role in the condition known as calcific aortic valve disease (CAVD). Here, we hypothesized that the AGE-LDL/RAGE axis could also be involved in the pathophysiological mechanism of CAVD. METHODS: Human aortic valve interstitial cells (HAVICs) were stimulated with AGE-LDL following pre-treatment with or without interleukin 37 (IL-37). Low-density lipoprotein receptor deletion (Ldlr-/-) hamsters were randomly allocated to chow diet (CD) group and high carbohydrate and high fat diet (HCHFD) group. RESULTS: AGE-LDL levels were significantly elevated in patients with CAVD and in a hamster model of aortic valve calcification. Our in vitro data further demonstrated that AGE-LDL augmented the expression of intercellular cell adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6) and alkaline phosphatase (ALP) in a dose-dependent manner through NF-κB activation, which was attenuated by nuclear factor kappa-B (NF-κB) inhibitor Bay11-7082. The expression of RAGE was augmented in calcified aortic valves, and knockdown of RAGE in HAVICs attenuated the AGE-LDL-induced inflammatory and osteogenic responses as well as NF-κB activation. IL-37 suppressed inflammatory and osteogenic responses and NF-κB activation in HAVICs. The vivo experiment also demonstrate that supplementation with IL-37 inhibited valvular inflammatory response and thereby suppressed valvular osteogenic activities. CONCLUSIONS: AGE-LDL promoted inflammatory responses and osteogenic differentiation through RAGE/NF-κB pathway in vitro and aortic valve lesions in vivo. IL-37 suppressed the AGE-LDL-induced inflammatory and osteogenic responses in vitro and attenuated aortic valve lesions in a hamster model of CAVD.

Oral health status and associated factors among 12 to 15-year-old Chinese adolescents in Southeast China: A cross-sectional study.

This study assessed oral health conditions and associated factors (including sociodemographic characteristics and self-reported oral health-related behaviors) among Chinese adolescents. This cross-sectional study enrolled 3840 adolescents aged 12 to 15 years from 12 middle schools in Foshan, Southeast China, in 2016, using multistage, stratified cluster sampling. Participants underwent a clinical oral examination and completed a questionnaire. The prevalence of dental caries, probe bleeding, and calculus was 37.6%, 46.2%, and 39.7%, respectively; the mean decayed/missed/filled teeth index was 0.86 ±â€…1.58. A mean of 2.09 ±â€…3.65 and 1.85 ±â€…3.52 teeth showed probe bleeding and calculus, respectively. Only 0.3% and 0.1% of adolescents aged 15 years had periodontal pockets (depth ≥ 4 mm) and attachment loss, respectively, which were most common in tooth positions 46 and 36 (Federation Dentaire International 2-digit system). Regarding oral health-related behavior, 49.1% of the participants failed to brush their teeth at least twice daily, 98.5% never or rarely used dental floss, and 58.7% reported middle-high frequency sugar consumption. Older age, female, administrative region, maternal education lower than university, brushing teeth less than twice daily, flossing less than once daily, and frequent sugar consumption were significant risk factors of caries. Older age, female, administrative region, brushing less than twice daily, and flossing less than once daily significantly increased periodontal risk. Despite the overall low prevalence of adverse dental conditions among adolescents in Foshan, their oral hygiene habits were undeveloped. Thus, their identified risk factors need close monitoring, and families, schools, communities, and the government should jointly promote adolescents' oral health.

Targeted inhibition of PTPN22 is a novel approach to alleviate osteogenic responses in aortic valve interstitial cells and aortic valve lesions in mice.

BACKGROUND: Calcific aortic valve disease (CAVD) is the most prevalent valvular disease and has high morbidity and mortality. CAVD is characterized by complex pathophysiological processes, including inflammation-induced osteoblastic differentiation in aortic valve interstitial cells (AVICs). Novel anti-CAVD agents are urgently needed. Protein tyrosine phosphatase nonreceptor type 22 (PTPN22), an intracellular nonreceptor-like protein tyrosine phosphatase, is involved in several chronic inflammatory diseases, including rheumatoid arthritis and diabetes. However, it is unclear whether PTPN22 is involved in the pathogenesis of CAVD. METHODS: We obtained the aortic valve tissue from human and cultured AVICs from aortic valve. We established CAVD mice model by wire injury. Transcriptome sequencing, western bolt, qPCR, and immunofluorescence were performed to elucidate the molecular mechanisms. RESULTS: Here, we determined that PTPN22 expression was upregulated in calcific aortic valve tissue, AVICs treated with osteogenic medium, and a mouse model of CAVD. In vitro, overexpression of PTPN22 induced osteogenic responses, whereas siRNA-mediated PTPN22 knockdown abolished osteogenic responses and mitochondrial stress in the presence of osteogenic medium. In vivo, PTPN22 ablation ameliorated aortic valve lesions in a wire injury-induced CAVD mouse model, validating the pathogenic role of PTPN22 in CAVD. Additionally, we discovered a novel compound, 13-hydroxypiericidin A 10-O-α-D-glucose (1 → 6)-ß-D-glucoside (S18), in a marine-derived Streptomyces strain that bound to PTPN22 with high affinity and acted as a novel inhibitor. Incubation with S18 suppressed osteogenic responses and mitochondrial stress in human AVICs induced by osteogenic medium. In mice with aortic valve injury, S18 administration markedly alleviated aortic valve lesions. CONCLUSION: PTPN22 plays an essential role in the progression of CAVD, and inhibition of PTPN22 with S18 is a novel option for the further development of potent anti-CAVD drugs. Therapeutic inhibition of PTPN22 retards aortic valve calcification through modulating mitochondrial dysfunction in AVICs.

Activation of Piezo1 promotes osteogenic differentiation of aortic valve interstitial cell through YAP-dependent glutaminolysis.

Hemodynamic overload and dysregulation of cellular metabolism are involved in development of calcific aortic valve disease (CAVD). However, how mechanical stress relates to metabolic changes in CAVD remains unclear. Here, we show that Piezo1, a mechanosensitive ion channel, regulated glutaminase 1 (GLS1)-mediated glutaminolysis to promote osteogenic differentiation of valve interstitial cells (VICs). In vivo, two models of aortic valve stenosis were constructed by ascending aortic constriction (AAC) and direct wire injury (DWI). Inhibition of Piezo1 and GLS1 in these models respectively mitigated aortic valve lesion. In vitro, Piezo1 activation induced by Yoda1 and oscillatory stress triggered osteogenic responses in VICs, which were prevented by Piezo1 inhibition or knockdown. Mechanistically, Piezo1 activation promoted calcium-dependent Yes-associated protein (YAP) activation. YAP modulated GLS1-mediated glutaminolysis, which enhanced osteogenic differentiation through histone acetylation of runt-related transcription factor 2 (RUNX2) promoters. Together, our work provided a cross-talk between mechanotransduction and metabolism in the context of CAVD.

Hesperetin, a Promising Dietary Supplement for Preventing the Development of Calcific Aortic Valve Disease.

BACKGROUND: No effective therapeutic agents for calcific aortic valve disease (CAVD) are available currently. Dietary supplementation has been proposed as a novel treatment modality for various diseases. As a flavanone, hesperetin is widely abundant in citrus fruits and has been proven to exert protective effects in multiple diseases. However, the role of hesperetin in CAVD remains unclear. METHODS: Human aortic valve interstitial cells (VICs) were isolated from aortic valve leaflets. A mouse model of aortic valve stenosis was constructed by direct wire injury (DWI). Immunoblotting, immunofluorescence staining, and flow cytometry were used to investigate the roles of sirtuin 7 (Sirt7) and nuclear factor erythroid 2-related factor 2 (Nrf2) in hesperetin-mediated protective effects in VICs. RESULTS: Hesperetin supplementation protected the mice from wire-injury-induced aortic valve stenosis; in vitro, hesperetin inhibited the lipopolysaccharide (LPS)-induced activation of NF-κB inflammatory cytokine secretion and osteogenic factors expression, reduced ROS production and apoptosis, and abrogated LPS-mediated injury to the mitochondrial membrane potential and the decline in the antioxidant levels in VICs. These benefits of hesperetin may have been obtained by activating Nrf2-ARE signaling, which corrected the dysfunctional mitochondria. Furthermore, we found that hesperetin could directly bind to Sirt7 and that the silencing of Sirt7 decreased the effects of hesperetin in VICs and potently abolished the ability of hesperetin to increase Nrf2 transcriptional activation. CONCLUSIONS: Our work demonstrates that hesperetin plays protective roles in the aortic valve through the Sirt7-Nrf2-ARE axis; thus, hesperetin might be a potential dietary supplement that could prevent the development of CAVD.

Marine-Derived Piericidin Diglycoside S18 Alleviates Inflammatory Responses in the Aortic Valve via Interaction with Interleukin 37.

Calcific aortic valve disease (CAVD) is a valvular disease frequently in the elderly individuals that can lead to the valve dysfunction. Osteoblastic differentiation of human aortic valve interstitial cells (HAVICs) induced by inflammation play a crucial role in CAVD pathophysiological processes. To date, no effective drugs for CAVD have been established, and new agents are urgently needed. Piericidin glycosides, obtained from a marine-derived Streptomyces strain, were revealed to have regulatory effects on mitochondria in previous studies. Here, we discovered that 13-hydroxypiericidin A 10-O-α-D-glucose (1→6)-ß-D-glucoside (S18), a specific piericidin diglycoside, suppresses lipopolysaccharide- (LPS) induced inflammatory responses of HAVICs by alleviating mitochondrial stress in an interleukin (IL)-37-dependent manner. Knockdown of IL-37 by siRNA not only exaggerated LPS-induced HAVIC inflammation and mitochondrial stress but also abrogated the anti-inflammatory effect of S18 on HAVICs. Moreover, S18 alleviated aortic valve lesions in IL-37 transgenic mice of CAVD model. Microscale thermophoresis (MST) and docking analysis of five piericidin analogues suggested that diglycosides, but not monoglycosides, exert obvious IL-37-binding activity. These results indicate that S18 directly binds to IL-37 to alleviate inflammatory responses in HAVICs and aortic valve lesions in mice. Piericidin diglycoside S18 is a potential therapeutic agent to prevent the development of CAVD.

4-Octyl itaconate suppresses the osteogenic response in aortic valvular interstitial cells via the Nrf2 pathway and alleviates aortic stenosis in mice with direct wire injury.

Calcific aortic valve disease (CAVD) is the most prevalent valvular heart disease in older individuals, but there is a lack of drug treatment. The cellular biological mechanisms of CAVD are still unclear. Oxidative stress and endoplasmic reticulum stress (ER stress) have been suggested to be involved in the progression of CAVD. Many studies have demonstrated that 4-octyl itaconate (OI) plays beneficial roles in limiting inflammation and oxidative injury. However, the potential role of OI in CAVD has not been thoroughly explored. Thus, we investigated OI-mediated modulation of ROS generation and endoplasmic reticulum stress to inhibit osteogenic differentiation in aortic valve interstitial cells (VICs). In our study, calcified aortic valves showed increased levels of ER stress and superoxide anion, as well as abnormal expression of Hmox1 and NQO1. In VICs, OI activated the Nrf2 signaling cascade and contributed to Nrf2 stabilization and nuclear translocation, thus augmenting the expression of genes downstream of Nrf2 (Hmox1 and NQO1). Moreover, OI ameliorated osteogenic medium (OM)-induced ROS production, mitochondrial ROS levels and the loss of mitochondrial membrane potential in VICs. Furthermore, OI attenuated the OM-induced upregulation of ER stress markers, osteogenic markers and calcium deposition, which were blocked by the Nrf2-specific inhibitor ML385. Interestingly, we found that OM-induced ER stress and osteogenic differentiation were ROS-dependent and that Hmox1 silencing triggered ROS production, ER stress and elevated osteogenic activity, which were inhibited by NAC. Overexpression of NQO1 mediated by adenovirus vectors significantly suppressed OM-induced ER stress and osteogenic markers. Collectively, these results showed the anti-osteogenic effects of OI on AVICs by regulating the generation of ROS and ER stress by activating the Nrf2 signaling pathway. Furthermore, OI alleviated aortic stenosis in a mouse model with direct wire injury. Due to its antioxidant properties, OI could be a potential drug for the prevention and/or treatment of CAVD.

Efficacy and safety of antithrombotic therapy with non-vitamin K antagonist oral anticoagulants after transcatheter aortic valve replacement: a systematic review and meta-analysis.

OBJECTIVE: A meta-analysis was performed to compare the efficacy and safety of antithrombotic therapy with non-vitamin K antagonist oral anticoagulants (NOACs) versus standard care in patients after successful transcatheter aortic valve replacement (TAVR). METHODS: A systematic search of PubMed, Cochrane Central Register of Controlled Trials, and EMBASE databases and ClinicalTrials.gov website (through 21 October 2020) was performed. Risk ratios (RRs) with 95% confidence intervals (CIs) for all outcomes were calculated using random-effects models. RESULTS: Twelve studies (two studies were randomized controlled trials) comprising 6943 patients were included (5299 had indications for oral anticoagulation (OAC) and 1644 had none). No significant differences were found between NOACs and the standard care in the incidences of all stroke, a composite endpoint, and major/life-threatening bleeding. NOACs were associated with lower all-cause mortality than vitamin K antagonists (VKAs) in post-TAVR patients with indications for OAC after more than 1 year of follow-up [RR = 0.64; 95% CI, (0.42, 0.96); p = 0.03], whereas NOACs exhibited poor outcomes than antiplatelet therapy (APT) in patients without indications for OAC [RR = 1.66; 95% CI, (1.12, 2.45); p = 0.01]. In the prevention of valve thrombosis, NOACs and VKAs were not significantly different in patients with indications for OAC [RR = 0.66; 95% CI, (0.24, 1.84); p = 0.43], whereas NOACs were better than APT in patients without indications for OAC [RR = 0.19; 95% CI, (0.04, 0.83); p = 0.03]. CONCLUSIONS: In patients with indications for OAC, post-TAVR antithrombotic therapy with NOACs was more favorable due to its lower all-cause mortality after more than 1 year of follow-up. In those without indications for OAC, NOACs presented poorer outcomes due to its higher all-cause mortality.

Patient value co-creation behavior scale based on the DART model.

OBJECTIVES: Value co-creation focuses on customer participation and co-creates value with suppliers. Patients' support and cooperation can improve the quality of medical care. Value creation is closely related to participants' behavior. The DART (Dialogue, Access, Risk assessment, Transparency) model is widely used in commercial research because it defines and classifies value co-creation behavior clearly and systematically. However, there is little research using the DART model in the field of health care. This study aimed to develop and evaluate the psychometric properties of a scale measuring patient value co-creation behavior based on the DART model. STUDY DESIGN: The Delphi technique was used to determine the scale content with a panel of 17 experts. A cross-sectional survey was administered to 356 outpatients and inpatients of a hospital in Guangzhou, China. METHODS: Internal consistency reliability and composite reliability (CR) were used to estimate the scale's reliability. Validity was assessed using convergent and discriminant validity. RESULTS: Three rounds of expert consultation were completed before a final consensus was reached regarding scale content. The patient value co-creation behavior scale was composed of 23 items and 4 dimensions. The overall Cronbach's α was 0.934, and the CRs of the 4 DART dimensions were 0.843, 0.872, 0.911, and 0.884, respectively, showing satisfactory reliability. The average variance extracted ranged from 0.473 to 0.659, and the χ2 difference between constrained and free models was significant, indicating convergent and discriminant validity. CONCLUSIONS: The scale exhibited acceptable reliability and validity and could serve as an evaluation tool for patient value co-creation behavior.

Interleukin 37 Suppresses M1 Macrophage Polarization Through Inhibition of the Notch1 and Nuclear Factor Kappa B Pathways.

Macrophage-orchestrated chronic inflammation plays an important role in cardiovascular disease, including accelerating the development of calcific aortic valve disease (CAVD). M1 and M2 macrophage polarization imbalances can alter intensity of inflammatory responses. Recombinant human interleukin 37 (IL-37) could be involved in regulating immune cell function to attenuate inflammation. This study aimed to identify IL-37 specifically modulates M1 polarization and investigate the underlying mechanism. Compared with normal valves, there are more M1 macrophages accumulation and less IL-37 expression in calcific aortic valves, which may indicate a negative relationship between IL-37 and M1 polarization. THP-1 cells could differentiate into resting macrophages with phorbol-12-myristate-13-acetate (PMA) and then polarize into M1 macrophages following treatment with lipopolysaccharide (LPS) and interferon gamma (IFN-γ). In vitro, recombinant human IL-37 attenuated the expression of inducible nitric oxide synthase (iNOS), CD11c, IL-6 and monocyte chemoattractant protein 1 (MCP-1) in M1 but augmented the expression of CD206 and IL-10 in M2. The suppression of M1 polarization was associated with the inhibition of the activation of the nuclear factor kappa B (NF-κB) and Notch1 signaling pathways. These results demonstrated that IL-37 inhibits the macrophages polarizing into M1 type via the inhibition of the Notch1 and nuclear factor kappa B pathways. In summary, IL-37 could be a potential therapeutic candidate for progressive CAVD by modulating M1 polarization and its orchestrated inflammation.

The Reliability and Validity of Short Form-12 Health Survey Version 2 for Chinese Older Adults.

BACKGROUND: We assessed the information regarding the psychometric properties of the Short Form-12 Health Survey Version 2 (SF-12v2) among older adults in China. METHODS: A cross-sectional study was conducted on a stratified representative sample of older adults (≥60 years) residing in community and nursing home settings in 2017-18. Reliability was estimated using the internal consistency method. Validity was assessed using convergent and discriminant validity checks, factor analyses (including both exploratory and confirmatory factor analyses [EFA and CFA]), and "known groups" construct validity. RESULTS: The final sample comprised 1000 older adults (451 community-dwelling and 549 institutional). Cronbach's α was 0.81 for the Physical Component Summary (PCS) and 0.83 for the Mental Component Summary (MCS), showing satisfactory internal consistency for both. Most items were strongly correlated with their represented component (Spearman's correlation coefficient: 0.62-0.87), although the correlation of SF items with PCS was a bit stronger than that with MCS. A two-factor structure (physical and mental health) indicated by EFA jointly accounted for 68.50% of the variance and presented adequate goodness-of-fit indices (GFI=0.98, AGFI=0.92, RMSEA=0.08, 90% Cl RMSEA=0.06 to 0.11, NFI=0.98, and CFI=0.98) in CFA. Known-groups comparison showed that SF-12v2 summary scores did well in differentiating subgroups of older adults by age, marital status, and self-reported health problems (P≤0.05). CONCLUSION: SF-12v2 is a reliable and valid health-related quality of life instrument for Chinese older adults that works equally well with older adults under institutional care and community-based home care models.

Health-related quality of life and related factors among elderly persons under different aged care models in Guangzhou, China: a cross-sectional study.

PURPOSE: To analyze health-related quality of life (HRQoL) and related factors among elderly persons receiving community-based home care and institutional care in Guangzhou, a large city of mainland China. METHODS: A representative sample of 1600 subjects aged 60 years and over residing in communities and nursing homes was randomly selected through stratified sampling. The 12-item Short Form Health Survey version 2 (SF-12v2) was used to assess HRQoL. RESULTS: In total, 1014 elderly persons under different aged care models responded to the survey (response rate 63.4%) and 1000 were eligible for data analyses. Compared with the elderly receiving community-based home care or private institutional care, those in public institutional care had the lowest scores on the physical component summary (PCS, 36.89 ± 10.44) and the mental component summary (MCS, 47.16 ± 11.14). Number of chronic diseases, loneliness, and age were the most common significant factors (P < 0.05) affecting PCS and MCS. The interaction term between aged care model and number of chronic diseases significantly affected PCS (ß = - 0.165, P < 0.05), indicating a stronger association between these factors for participants receiving community-based home care than institutional care. The interaction term between aged care model and loneliness had a significant effect on MCS (ß = 0.189, P < 0.05), indicating a weaker association between loneliness and MCS for participants receiving community-based home care. CONCLUSIONS: This study found poor HRQoL among the elderly in Guangzhou. The main factors associated with the physical and mental HRQoL of elderly persons included number of chronic diseases, loneliness, age, and education level. It also revealed the moderating effects of aged care model on HRQoL, suggesting specific health management strategies for elderly in community-based home care and institutional care, respectively.

Association between homocysteine levels and calcific aortic valve disease: a systematic review and meta-analysis.

Previous studies have reported inconsistent results regarding the association between homocysteine (Hcy) levels and calcific aortic valve disease (CAVD). We investigate the association between Hcy levels in patients with CAVD and controls by conducting a systematic review and meta-analysis. We conducted a systematic search of studies published prior to the end of March 2017 in the PubMed, Embase, Web of Science, Cochrane Central Register of Controlled Trials and the Chinese Biomedical Literature databases. Eligible studies evaluating plasma Hcy levels in CAVD patients and controls were identified by two independent investigators. Standardized mean difference (SMD) and the corresponding 95% confidence intervals (95% CIs) were estimated using the random-effects model. Ten studies involving 6349 participants were included. Pooled analysis demonstrated that Hcy levels were significantly elevated in patients with CAVD compared with controls (pooled SMD: 0.57, 95% CI: 0.36-0.79). This elevation was more obvious in American and Asian populations than in Turkish populations. Furthermore, Hcy levels were significantly elevated in patients with mild-to-moderate CAVD and severe CAVD. Our results demonstrate that CAVD is associated with elevated Hcy levels.

Long-term compulsive exercise reduces the rewarding efficacy of 3,4-methylenedioxymethamphetamine.

Although exercise has been known to regulate brain plasticity, its impact on psychostimulant reward and the associated mesolimbic dopamine system remained scarcely explored. A psychostimulant, 3,4-methylenedioxymethamphetamine (MDMA), is currently a worldwide abused drug of choice. We decided to examine the modulating effects of long-term, compulsive treadmill exercise on the hedonic value of MDMA in male C57BL/6J mice. MDMA-induced conditioned place preference (CPP) was used as a behavioral paradigm to indicate the reward efficacy of MDMA. We observed that sedentary control mice all demonstrated reliable MDMA-induced CPP with our conditioning protocol. Interestingly, pre-exposure to a treadmill exercise decreased the later MDMA-induced CPP in a running period-dependent manner. Specifically, mice undergoing a 12-week treadmill running exercise did not exhibit any approaching bias toward the MDMA-associated compartment in this CPP paradigm. Twelve weeks of treadmill running did not alter peripheral metabolism of MDMA 30min following single intraperitoneal injection of MDMA (3mg/kg). We further used microdialysis technique to study the underlying mechanisms for the impaired MDMA reward produced by the12-week exercise pre-exposure. We found that acute MDMA-stimulated dopamine release in nucleus accumbens was abolished in the exercised mice, whereas an obvious elevation of accumbal dopamine release was observed in sedentary control mice. Finally, the 12-week exercise program did not alter the protein levels of primary dopamine receptors, vesicular or membrane transporters in this area. We conclude that the long-term, compulsive exercise is effective in curbing the reward efficacy of MDMA possibly via its direct effect on reversing the MDMA-stimulated dopamine release in nucleus accumbens.

Prenatal exposure of bupropion may enhance agitation, anxiety responses, and sensitivity to cocaine effects in adult mice.

Major depression and dysthymia afflict a proportion of gravid and breast-feeding women. These women are frequently recommended on antidepressants to relieve their symptoms even if the drug effects on fetal growth and postnatal development are not completely known. In a previous study, we reported that prenatal bupropion exposure seemed to enhance the hedonic value of cocaine in adult mice. This study was undertaken to examine the dose-related effects for prenatal bupropion exposure on the stress susceptibility, cocaine-associated reinforcing property, and cocaine-induced behavioral sensitization in adult mice. Our results showed that various doses (ranging 12.5-50 mg/kg) of prenatal bupropion administration at the third trimester of pregnancy did not affect body weight of the adult mice. Bupropion administration at 50 mg/kg enhanced both ambulatory and rearing responses in the open field test. Moreover, bupropion administration (at 25 and 50 mg/kg) significantly decreased the numbers in open arm entry in the elevated plus maze test. Furthermore, prenatal bupropion treatment appeared to facilitate the cocaine-induced place preference in a sex-dependent manner. Finally, prenatal bupropion exposure (at 25 and 50 mg/kg) accelerated and elevated the development of cocaine-induced sensitization in locomotor activity. While the antidepressant and smoking-curbing effects of bupropion have been addressed in literature, we suggest that prenatal bupropion exposure could run a risk of enhancing individual's agitation, stress susceptibility and cocaine stimulating propensity in adulthood.