Challenges and new opportunities for detecting endogenous opioid peptides in reward

The endogenous opioid peptide system, comprised of enkephalins, endorphins, dynorphins, and nociceptin, is a highly complex neurobiological system. Opioid peptides are derived from four precursor molecules and undergo several processing events yielding over 20 unique opioid peptides. This diversity together with lo

Forever-Fit Summer Camp: The Impact of a 6-Week Summer Healthy Lifestyle Day Camp on Anthropometric, Cardiovascular, and Physical Fitness Measures in Youth With Obesity

Pediatric obesity is a public health concern with lifestyle intervention as the first-line treatment. Forever-Fit Summer Camp (FFSC) is a 6-week summer day program offering physical activity, nutrition education, and well-balanced meals to youth at low cost. The aim of the study was to assess the efficacy of this program that does not emphasize weight loss rather emphasizes healthy behaviors on body mass index, cardiovascular and physical fitness. Methods: The inclusion criteria were adolescents between 8 and 12 years and body mass index (BMI) ≥85th percentile. The data were collected at baseline and week 6 (wk-6) and was analyzed for 2013-2018 using paired-sample t tests. Results: The participants' (N = 179) average age was 10.6 ± 1.6 years with a majority of females (71%) and black race/ethnicity (70%). At wk-6, BMI and waist circumference decreased by 0.8 ± 0.7 kg/m2 and 1.0 ± 1.3 in, respectively. Resting heart rate, diastolic and systolic blood pressure decreased by 8.5 ± 11.0 bpm, 6.3 ± 8.8 mmHg, and 6.4 ± 10.1 mmHg, respectively. The number of pushups, curl-ups, and chair squats were higher by 5.8 ± 7.5, 6.7 ± 9.1, and 7.7 ± 8.5, respectively. Conclusion: The FFSC is efficacious for improving BMI, cardiovascular, and physical fitness in the short term. The effect of similar episodic efforts that implement healthy lifestyle modifications throughout the school year should be investigated

Ontogenetic oxycodone exposure affects early life communicative behaviors, sensorimotor reflexes, and weight trajectory in mice

Nationwide, opioid misuse among pregnant women has risen four-fold from 1999 to 2014, with commensurate increase in neonates hospitalized for neonatal abstinence syndrome (NAS). NAS occurs when a fetus exposed to opioid

Molecular Mechanism of Action of Antimalarial Benzoisothiazolones: Species-Selective Inhibitors of the Plasmodium spp. MEP Pathway enzyme, IspD

The methylerythritol phosphate (MEP) pathway is an essential metabolic pathway found in malaria parasites, but absent in mammals, making it a highly attractive target for the discovery of novel and selective antimalarial therapies. Using high-throughput screening, we have identified 2-phenyl benzo[d]isothiazol-3(2H)-ones as species-selective inhibitors of Plasmodium spp. 2-C-methyl-D-erythritol-4-phosphate cytidyltransferase (IspD), the third catalytic enzyme of the MEP pathway. 2-Phenyl benzo[d]isothiazol-3(2H)-ones display nanomolar inhibitory activity against P. falciparum and P. vivax IspD and prevent the growth of P. falciparum in culture, with EC50 values below 400 nM. In silico modeling, along with enzymatic, genetic and crystallographic studies, have established a mechanism-of-action involving initial non-covalent recognition of inhibitors at the IspD binding site, followed by disulfide bond formation through attack of an active site cysteine residue on the benzo[d]isothiazol-3(2H)-one core. The species-selective inhibitory activity of these small molecules against Plasmodium spp. IspD and cultured parasites suggests they have potential as lead compounds in the pursuit of novel drugs to treat malaria

Short-Term Change in Measures of Glycemia in Obese Youth Meeting Criteria for Prediabetes: A Retrospective Chart Review

Background: The prevalence of youth diagnosed with prediabetes is increasing, yet there is a lack of guidelines on how to manage this condition clinically. Objectives: The aim was to determine the short-term outcomes of patients referred with prediabetes and to determine predictors of worsening dysglycemia in youth. Study Design: This is a retrospective chart review of patients referred to our Youth Diabetes Prevention Clinic (YDPC) with laboratory tests indicating an increased risk for type 2 diabetes (T2D). We defined glycemic categories by HbA1c with normoglycemia as HbA1c <5.7%, prediabetes I (P1) as HbA1c 5.7 to <6.0%, and prediabetes II (P2) as HbA1c 6.0 to <6.5%. We compared HbA1c at the time of referral (screening HbA1c) and at the YDPC visit (YDPC HbA1c) to assess for improvement or worsening. Multinomial logistic regression was used to assess predictors of prediabetes. Results: Among 562 patients seen, 336 had both screening and YDPC HbA1c values. Race (p < 0.001) and screening glycemic category (p < 0.001) were significantly associated with dysglycemia at the YDPC visit, while sex (p = 0.50), BMI z-score change (p = 0.27), and days from referral (p = 0.83) were not. As compared to those who reverted to normoglycemia, patients with prediabetes at YDPC were 7 times more likely to have a higher screening HbA1c (both P1 and P2). The majority of patients referred with prediabetes had lower HbA1c at the YDPC (75.4–82.6%). Conclusion: Patients with screening HbA1c <6% might benefit from a 4-month follow-up at primary care while recommending lifestyle changes. Patients of minority race and screening HbA1c ≥6% are more likely to have a persistent elevation of HbA1c

Intervention effects of Ganoderma lucidum spores on epileptiform discharge hippocampal neurons and expression of Neurotrophin-4 and N-Cadherin

Epilepsy can cause cerebral transient dysfunctions. Ganoderma lucidum spores (GLS), a traditional Chinese medicinal herb, has shown some antiepileptic effects in our previous studies. This was the first study of the effects of GLS on cultured primary hippocampal neurons, treated with Mg2+ free medium. This in vitro model of epileptiform discharge hippocampal neurons allowed us to investigate the anti-epileptic effects and mechanism of GLS activity. Primary hippocampal neurons from <1 day old rats were cultured and their morphologies observed under fluorescence microscope. Neurons were confirmed by immunofluorescent staining of neuron specific enolase (NSE). Sterile method for GLS generation was investigated and serial dilutions of GLS were used to test the maximum non-toxic concentration of GLS on hippocampal neurons. The optimized concentration of GLS of 0.122 mg/ml was identified and used for subsequent analysis. Using the in vitro model, hippocampal neurons were divided into 4 groups for subsequent treatment i) control, ii) model (incubated with Mg2+ free medium for 3 hours), iii) GLS group I (incubated with Mg2+ free medium containing GLS for 3 hours and replaced with normal medium and incubated for 6 hours) and iv) GLS group II (neurons incubated with Mg2+ free medium for 3 hours then replaced with a normal medium containing GLS for 6 hours). Neurotrophin-4 and N-Cadherin protein expression were detected using Western blot. The results showed that the number of normal hippocampal neurons increased and the morphologies of hippocampal neurons were well preserved after GLS treatment. Furthermore, the expression of neurotrophin-4 was significantly increased while the expression of N-Cadherin was decreased in the GLS treated group compared with the model group. This data indicates that GLS may protect hippocampal neurons by promoting neurotrophin-4 expression and inhibiting N-Cadherin expression

Integrating Diabetes Prevention Education Among Teenagers Involved in Summer Employment: Encouraging Environments for Health in Adolescence (ENHANCE)

Type 2 diabetes (T2D) in youth is a growing healthcare and public health concern. It is costly, and youth suffer from disabling and deadly comorbid conditions at a faster pace than adult onset. However, T2D is preventable. The population of obese youth at greatest risk for T2D is of minority race/ethnicity and socioeconomically disadvantaged background, which creates barriers to health promoting lifestyles. Despite being the first line of prevention efforts for T2D, efficacious behavioral lifestyle interventions are still lacking at the community level. During the summers of 2016 and 2017, a study integrated obesity and diabetes prevention health education into TeenWorks summer employment program at Indy Urban Acres in Indianapolis, Indiana. Results were analyzed using paired sample t-tests. Participants (N = 168) had a mean age of 15.8 ± 0.7 years, 61% female, 13% Hispanic, 80% Black. By the end of the intervention, physical activity (p = 0.000) and prevention knowledge (p = 0.000) were significantly higher. Dietary intake (p = 0.204), self-efficacy (p = 0.58), food insecurity (p = 0.058) and depression screening scores (p = 0.809) were not significantly different. In light of the continuing childhood obesity epidemic and increasing prevalence of prediabetes and T2D in youth, there is a pressing need to understand and reduce barriers to obesity and diabetes prevention in high-risk populations. This study demonstrated the feasibility of integrating obesity and T2D prevention health education into a teen summer employment program

Alternating hemiplegia of childhood: evolution over time and mouse model corroboration

Alternating hemiplegia of childhood is a rare neurodevelopmental disorder caused by ATP1A3 mutations. Some evidence for disease progression exists, but there are few systematic analyses. Here, we evaluate alternating hemiplegia of childhood progression in humans and in the D801N knock-in alternating hemiplegia of childhood mouse, Mashlool, model. This study performed an ambidirectional (prospective and retrospective data) analysis of an alternating hemiplegia of childhood patient cohort (n = 42, age 10.24 ± 1.48 years) seen at one US centre. To investigate potential disease progression, we used linear mixed effects models incorporating early and subsequent visits, and Wilcoxon Signed Rank test comparing first and last visits. Potential early-life clinical predictors were determined via multivariable regression. We also compared EEG background at first encounter and at last follow-up. We then performed a retrospective confirmation study on a multicentre cohort of alternating hemiplegia of childhood patients from France (n = 52). To investigate disease progression in the Mashlool mouse, we performed behavioural testing on a cohort of Mashlool- mice at prepubescent and adult ages (n = 11). Results: US patients, over time, demonstrated mild worsening of non-paroxysmal disability index scores, but not of paroxysmal disability index scores. Increasing age was a predictor of worse scores: P < 0.0001 for the non-paroxysmal disability index, intellectual disability scale and gross motor scores. Earliest non-paroxysmal disability index score was a predictor of last visit non-paroxysmal disability index score (P = 0.022), and earliest intellectual disability score was a predictor of last intellectual disability score (P = 0.035). More patients with EEG background slowing were noted at last follow-up as compared to initial (P = 0.015). Similar worsening of disease with age was also noted in the French cohort: age was a significant predictor of non-paroxysmal disability index score (P = 0.001) and first and last non-paroxysmal disability index score scores significantly differed (P = 0.002). In animal studies, adult Mashlool mice had, as compared to younger Mashlool mice, (i) worse balance beam performance; (ii) wider base of support; (iii) higher severity of seizures and resultant mortality; and (iv) no increased predisposition to hemiplegic or dystonic spells. In conclusion, (i) non-paroxysmal alternating hemiplegia of childhood manifestations show, on average over time, progression associated with severity of early-life non-paroxysmal disability and age. (ii) Progression also occurs in Mashlool mice, confirming that ATP1A3 disease can lead to age-related worsening. (iii) Clinical findings provide a basis for counselling patients and for designing therapeutic trials. Animal findings confirm a mouse model for investigation of underlying mechanisms of disease progression, and are also consistent with known mechanisms of ATP1A3-related neurodegeneration

Functional genome-wide siRNA screen identifies KIAA0586 as mutated in Joubert syndrome

Defective primary ciliogenesis or cilium stability forms the basis of human ciliopathies, including Joubert syndrome (JS), with defective cerebellar vermis development. We performed a high-content genome wide siRNA screen to identify genes regulating ciliogenesis as candidates for JS. We analyzed results with a supervised learning approach, using SYSCILIA gold standard, Cildb3.0, a centriole siRNA screen and the GTex project, identifying 591 likely candidates. Intersection of this data with whole exome results from 145 individuals with unexplained JS identified six families with predominantly compound heterozygous mutations in KIAA0586. A c.428del base deletion in 0.1% of the general population was found in trans with a second mutation in an additional set of 9 of 163 unexplained JS patients. KIAA0586 is an orthologue of chick Talpid3, required for ciliogenesis and sonic hedgehog signaling. Our results uncover a relatively high frequency cause for JS and contribute a list of candidates for future gene discoveries in ciliopathies

Clinical profile of patients with ATP1A3 mutations in Alternating Hemiplegia of Childhood\u2014a study of 155 patients

Background: Mutations in the gene ATP1A3 have recently been identified to be prevalent in patients with alternating hemiplegia of childhood (AHC2). Based on a large series of patients with AHC, we set out to identify the spectrum of different mutations within the ATP1A3 gene and further establish any correlation with phenotype. Methods: Clinical data from an international cohort of 155 AHC patients (84 females, 71 males; between 3 months and 52 years) were gathered using a specifically formulated questionnaire and analysed relative to the mutational ATP1A3 gene data for each patient. Results: In total, 34 different ATP1A3 mutations were detected in 85 % (132/155) patients, seven of which were novel. In general, mutations were found to cluster into five different regions. The most frequent mutations included: p.Asp801Asn (43 %; 57/132), p.Glu815Lys (16 %; 22/132), and p.Gly947Arg (11 %; 15/132). Of these, p.Glu815Lys was associated with a severe phenotype, with more severe intellectual and motor disability. p.Asp801Asn appeared to confer a milder phenotypic expression, and p.Gly947Arg appeared to correlate with the most favourable prognosis, compared to the other two frequent mutations. Overall, the comparison of the clinical profiles suggested a gradient of severity between the three major mutations with differences in intellectual (p = 0.029) and motor (p = 0.039) disabilities being statistically significant. For patients with epilepsy, age at onset of seizures was earlier for patients with either p.Glu815Lys or p.Gly947Arg mutation, compared to those with p.Asp801Asn mutation (p &lt; 0.001). With regards to the five mutation clusters, some clusters appeared to correlate with certain clinical phenotypes. No statistically significant clinical correlations were found between patients with and without ATP1A3 mutations. Conclusions: Our results, demonstrate a highly variable clinical phenotype in patients with AHC2 that correlates with certain mutations and possibly clusters within the ATP1A3 gene. Our description of the clinical profile of patients with the most frequent mutations and the clinical picture of those with less common mutations confirms the results from previous studies, and further expands the spectrum of genotype-phenotype correlations. Our results may be useful to confirm diagnosis and may influence decisions to ensure appropriate early medical intervention in patients with AHC. They provide a stronger basis for the constitution of more homogeneous groups to be included in clinical trial