Differential adeno-associated virus mediated gene transfer to sensory neurons following intrathecal delivery by direct lumbar puncture

<p>Abstract</p> <p>Background</p> <p>Neuronal transduction by adeno-associated viral (AAV) vectors has been demonstrated in cortex, brainstem, cerebellum, and sensory ganglia. Intrathecal delivery of AAV serotypes that transduce neurons in dorsal root ganglia (DRG) and spinal cord offers substantial opportunities to 1) further study mechanisms underlying chronic pain, and 2) develop novel gene-based therapies for the treatment and management of chronic pain using a non-invasive delivery route with established safety margins. In this study we have compared expression patterns of AAV serotype 5 (AAV5)- and AAV serotype 8 (AAV8)-mediated gene transfer to sensory neurons following intrathecal delivery by direct lumbar puncture.</p> <p>Results</p> <p>Intravenous mannitol pre-treatment significantly enhanced transduction of primary sensory neurons after direct lumbar puncture injection of AAV5 (rAAV5-GFP) or AAV8 (rAAV8-GFP) carrying the green fluorescent protein (GFP) gene. The presence of GFP in DRG neurons was consistent with the following evidence for primary afferent origin of the majority of GFP-positive fibers in spinal cord: 1) GFP-positive axons were evident in both dorsal roots and dorsal columns; and 2) dorsal rhizotomy, which severs the primary afferent input to spinal cord, abolished the majority of GFP labeling in dorsal horn. We found that both rAAV5-GFP and rAAV8-GFP appear to preferentially target large-diameter DRG neurons, while excluding the isolectin-B4 (IB4) -binding population of small diameter neurons. In addition, a larger proportion of CGRP-positive cells was transduced by rAAV5-GFP, compared to rAAV8-GFP.</p> <p>Conclusions</p> <p>The present study demonstrates the feasibility of minimally invasive gene transfer to sensory neurons using direct lumbar puncture and provides evidence for differential targeting of subtypes of DRG neurons by AAV vectors.</p

Extending the remit of evidence-based policing

Evidence-based policing (EBP) is an important strand of the UK’s College of Policing’s Police Education Qualifications Framework (PEQF), itself a component of a professionalisation agenda. This article argues that the two dominant approaches to EBP, experimental criminology and crime science, offer limited scope for the development of a comprehensive knowledge base for policing. Although both approaches share a common commitment to the values of science, each recognizes their limited coverage of policing topics. The fundamental difference between them is what each considers ‘best’ evidence. This article critically examines the generation of evidence by these two approaches and proposes an extension to the range of issues EBP should cover by utilizing a greater plurality of methods to exploit relevant research. Widening the scope of EBP would provide a broader foundational framework for inclusion in the PEQF and offers the potential for identifying gaps in the research, constructing blocks for knowledge building, and syllabus development in higher level police education

The GAIN Short Screener (GSS) as a Predictor of Future Arrest or Incarceration among Youth Presenting to Substance Use Disorder (SUD) Treatment

The National Institutes of Health (NIH) data harmonization project on existing measures ( www.phenx.org ) has recommended the Global Appraisal of Individual Needs (GAIN)–-Short Screener (GSS) as one of the most reliable, valid, efficient, and inexpensive general behavioral health screeners to quickly identify people with internalizing and externalizing mental health disorders, substance use disorders, and crime/violence problems. The present study examined how well the four GSS screeners and their sum predict future arrest or incarceration among individuals entering treatment for a substance use disorder. Using a cross-validation design, a diverse sample of 6,815 youth with substance use disorders was split into a development sample and a validation sample. Overall, results found the GSS's crime and violence screener (CVScr) and the substance disorder screener (SDScr) to be the two best predictors of arrest/incarceration within the 12 months following treatment intake. Additionally, we found that these screeners could be used to categorize individuals into three groups (low risk, moderate risk, high risk) and this simplified classification had good predictive validity (Area Under the Curve = 0.601). In sum, the GSS's predictive validity was similar to other instruments that have been developed to predict risk for recidivism; however, the GSS takes only a fraction of the time to collect (ie, approximately 2–3 minutes for just these two screeners)

Stress-Induced Cellular Clearance Is Mediated by the SNARE Protein ykt6 and Disrupted by α-Synuclein.

Age-related neurodegenerative disorders are characterized by a slow, persistent accumulation of aggregated proteins. Although cells can elicit physiological responses to enhance cellular clearance and counteract accumulation, it is unclear how pathogenic proteins evade this process in disease. We find that Parkinson's disease α-synuclein perturbs the physiological response to lysosomal stress by impeding the SNARE protein ykt6. Cytosolic ykt6 is normally autoinhibited by a unique farnesyl-mediated regulatory mechanism; however, during lysosomal stress, it activates and redistributes into membranes&nbsp;to preferentially promote hydrolase trafficking and enhance cellular clearance. α-Synuclein aberrantly binds and deactivates ykt6 in patient-derived neurons, thereby disabling the lysosomal stress response and facilitating protein accumulation. Activating ykt6 by small-molecule farnesyltransferase inhibitors restores lysosomal activity and reduces α-synuclein in patient-derived neurons and mice. Our findings indicate that α-synuclein creates a permissive environment for aggregate persistence by inhibiting regulated cellular clearance and provide a therapeutic strategy to restore protein homeostasis by harnessing SNARE activity