A New Photocaged Puromycin for an Efficient Labeling of Newly Translated Proteins in Living Neurons.

Monitoring newly synthesized proteins is becoming increasingly important to characterize proteome composition in regulatory networks. Puromycin is a peptidyl transfer inhibitor, widely used in cell biology for tagging newly synthesized proteins. Here, we report synthesis and application of an optimized puromycin carrying a photolabile protecting group as a powerful tool for tagging nascent proteins with high spatiotemporal resolution. The photocaged 7-N,N-(diethylaminocumarin-4-yl)-methoxycarbonyl-puromycin (DEACM-puromycin) was synthesized and compared with the previously developed 6-nitroveratryloxycarbonyl puromycin (NVOC-puromycin). The photochemical behavior as well as the effectiveness in controlling puromycylation in living hippocampal neurons using two-photon excitation is superior to the previously used NVOCpuromycin. We further report on the application of light-controlled puromycylation to visualize new translated proteins in neurons.

Comparison of Aberrant Behavior Checklist profiles across Prader-Willi syndrome, Down syndrome, and autism spectrum disorder.

Prader-Willi syndrome (PWS, OMIM # 176270) and Down syndrome (DS, OMIM #190685) are neurodevelopmental genetic disorders with higher rates of autism spectrum disorder (ASD). The Aberrant Behavior Checklist (ABC) is a caregiver rating scale that assesses maladaptive behaviors. Overlapping symptoms exist between PWS, DS, and ASD, including maladaptive behaviors. We aimed to evaluate ABC profiles between PWS, DS, and ASD alone (without known genetic syndrome). In addition, we hypothesized PWS and DS with a comorbid ASD positive screen or diagnosis would have similar ABC profiles to ASD alone. ABC data from the following cohorts were analyzed: PWS (Seattle Children's Hospital, n = 28, mean age = 12.8 ± 4.9 years; University of Florida, n = 35, mean age = 9.3 ± 7.1 years), DS (Johns Hopkins, n = 406, mean age = 8.1 ± 2.4 years), and ASD (University of Florida, n = 102, mean age = 10.8 ± 3.5 years). ASD alone had significantly higher ABC scores. Subgroups of PWS and DS with a comorbid ASD positive screen or diagnosis had similarities in scores with the ASD only group, with subscale patterns unique to each syndrome. The ABC indicated worse maladaptive behaviors in children with ASD, including those with genetic syndromes. Although more studies are needed to evaluate the utility and the accuracy of the ABC as a tool to screen for ASD in special populations, it may be a useful adjunct in screening those children with PWS or DS who need more in depth ASD evaluation.

Early intervention and the role of pediatricians.

Growing evidence supports the efficacious nature of early intervention (EI) services to children with developmental delays and their families. Pediatricians can play a crucial role in identifying developmental delays and referring families to EI services provided by each state, under Part C of the Individuals with Disabilities Education Act (IDEA). The goals of this article are to educate pediatricians about the benefits of EI services and resources available within their state to help facilitate family involvement in EI. By being involved in the EI process, pediatricians can work as partners with parents and multidisciplinary teams to provide seamless coordinated care to children and their families.

Research Brief: The Teen ADHD Workshop to Improve Adolescent ADHD Treatment Engagement.

Objective: Adolescents with ADHD are at risk of functional problems that may be mitigated by consistent ADHD treatment. This study pilots a brief intervention for adolescents with ADHD and their parents to increase treatment knowledge and family motivation to seek treatment. Method: The 3-hr curriculum was developed by a multidisciplinary team and included psychoeducation, goal setting, and motivational interviewing. Fifteen adolescents and 20 caregivers participated in the workshop, completed pre- and post-test assessments, and reported on acceptability. Results: Acceptability and satisfaction with the intervention were high. Perceived knowledge of ADHD increased post intervention; stigma was unchanged. Parents reported more acceptability of stimulant medications and less willingness to use special diets or cognitive games. Family feedback informed modifications to the curriculum. Conclusion: The Teen ADHD Workshop is a feasible and acceptable intervention to increase knowledge of ADHD and evidence-based treatments. Further research will evaluate effects on treatment participation.

Intellectual Disability and Psychotropic Medications.

CASE: Andrew is a 17-year-old male with trisomy 21, commonly known as Down syndrome, and accompanying severe intellectual disability who presents to your primary care office with his father for the first time to establish care and assistance with transition. Andrew has a history of a complete atrioventricular canal that was repaired as an infant and poorly controlled infantile spasms. Currently, he struggles with constipation, esophageal strictures, medullary nephrocalcinosis, urinary retention, sleep dysregulation, G-tube dependency, and hip dysplasia.Andrew walked at 11 to 12 years of age. Currently, he ambulates on his feet at home and in a wheelchair out in the community. He is nonverbal but can imprecisely sign for "more" and understands a few words. His father reports that his main concern is long-standing nonsuicidal self-injury (NSSI) and aggression. His self-injury consists of head banging against hard objects such as concrete floors and biting or scratching himself to the point of bleeding. Over the past 13 years, he has been prescribed over 10 different psychotropic medications, including various typical and atypical antipsychotics, selective serotonin reuptake inhibitors, benzodiazepines, mood stabilizers, and alpha agonists, all of which were discontinued because of the perception of undesirable side effects or lack of efficacy. His current medications include aripiprazole, olanzapine, levetiracetam, clorazepate, and trazodone. To rule out causes of irritability, you order a brain and spine magnetic resonance imaging, metabolic testing (for causes of NSSI such as Lesch-Nyhan), an autoimmune workup (for causes of pain or inflammation such as juvenile idiopathic arthritis), and hearing/vision testing, which are all normal. Previous testing by subspecialists (he is followed by gastroenterology, sleep medicine, orthopedics, nephrology, neurology, cardiology, and psychiatry) included normal renal ultrasound and no clear sources of gastrointestinal pain. However, key providers are spread among multiple institutions and do not regularly communicate.Andrew lives with his parents, who are highly educated and very dedicated to his health and wellness. His mother travels frequently for work, and his father is Andrew's full-time caregiver. Despite remaining ostensibly positive, his father reports significant caregiver burnout and fatigue.Over the next several months, Andrew continues to experience worsening NSSI necessitating medication changes despite active involvement in applied behavior analysis therapy. During this time, he presents to the emergency department multiple times for irritability and self-injury. On examination, he is aggressive, irritable, has bruises on his forehead and scratches on his skin, and has intermittent vertical gaze deviation that was noticeable to parents. The rest of his physical and neurological examination was unremarkable and revealed no asymmetry, clonus, hyperreflexia, or changes in muscle tone. While examining his extremities, joints, and abdomen, there was no obvious source of pain.What are your next steps? How would you support this family, both in the immediate management of his self-injury and long-term care needs for this medically and behaviorally complex adolescent?

Water-soluble Py-BIPS spiropyrans as photoswitches for biological applications.

The ultrafast photochemistry of a new spiropyran photoswitch (Py-BIPS) has been investigated, revealing many advantages in the application in water over the previously studied spiropyrans. Functionalized Py-BIPS derivatives are presented for the study of pH dependence, stability, toxicity, and the thermal and photochemical behavior on longer time scales in aqueous media using several spectroscopic methods. These investigations pave the way for the practical use of Py-BIPS derivatives as photoswitchable ligands of biomolecules.