The non-pathogenic parvovirus, adeno-associated virus (AAV), is an efficient vector for transgene expression in vivo and shows promise for treatment of brain disorders in clinical trials. Currently, there are more than 100 AAV serotypes identified that differ in the binding capacity of capsid proteins to specific cell surface receptors that can transduce different cell types and brain regions in the CNS. In the current study, multiple AAV serotypes expressing a GFP reporter (AAV1, AAV2/1, AAVDJ, AAV8, AAVDJ8, AAV9, AAVDJ9) were screened for their infectivity in both primary murine astrocyte and neuronal cell cultures. AAV2/1, AAVDJ8 and AAV9 were selected for further investigation of their tropism throughout different brain regions and cell types. Each AAV was administered to P0-neonatal mice via intracerebroventricular injections (ICV). Brains were then systematically analyzed for GFP expression at 3 or 6 weeks post-infection in various regions, including the olfactory bulb, striatum, cortex, hippocampus, substantia nigra (SN) and cerebellum. Cell counting data revealed that AAV2/1 infections were more prevalent in the cortical layers but penetrated to the midbrain less than AAVDJ8 and AAV9. Additionally, there were differences in the persistence of viral transgene expression amongst the three serotypes examined in vivo at 3 and 6 weeks post-infection. Because AAV-mediated transgene expression is of interest in neurodegenerative diseases such as Parkinson’s Disease, we examined the SN with microscopy techniques, such as CLARITY tissue transmutation, to identify AAV serotypes that resulted in optimal transgene expression in either astrocytes or dopaminergic neurons. AAVDJ8 displayed more tropism in astrocytes compared to AAV9 in the SN region. We conclude that ICV injection results in lasting expression of virally encoded transgene when using AAV vectors and that specific AAV serotypes are required to selectively deliver transgenes of interest to different brain regions in both astrocytes and neurons.

The orphan nuclear receptor Nurr1 (also called nuclear receptor-4A2) regulates inflammatory gene expression in glial cells, as well as genes associated with homeostatic and trophic function in dopaminergic neurons. Despite these known functions of Nurr1, an endogenous ligand has not been discovered. We postulated that the activation of Nurr1 would suppress the activation of glia and thereby protect against loss of dopamine (DA) neurons after subacute lesioning with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Our previous studies have shown that a synthetic Nurr1 ligand, 1,1-bis(3′-indolyl)-1-(p-chlorophenyl)methane (C-DIM12), suppresses inflammatory gene expression in primary astrocytes and induces a dopaminergic phenotype in neurons. Pharmacokinetic analysis of C-DIM12 in mice by liquid chromatography-mass spectrometry demonstrated that approximately three times more compound concentrated in the brain than in plasma. Mice treated with four doses of MPTP + probenecid over 14 days were monitored for neurobehavioral function, loss of dopaminergic neurons, and glial activation. C-DIM12 protected against the loss of DA neurons in the substantia nigra pars compacta and DA terminals in the striatum, maintained a ramified phenotype in microglia, and suppressed activation of astrocytes. In vitro reporter assays demonstrated that C-DIM12 was an effective activator of Nurr1 transcription in neuronal cell lines. Computational modeling of C-DIM12 binding to the three-dimensional structure of human Nurr1 identified a high-affinity binding interaction with Nurr1 at the coactivator domain. Taken together, these data suggest that C-DIM12 is an activator of Nurr1 that suppresses glial activation and neuronal loss in vivo after treatment with MPTP, and that this receptor could be an efficacious target for disease modification in individuals with Parkinson’s disease and related disorders.

Background: The growing focus on subjective patient experiences has created an increase in popularity for physician rating websites. The purpose of this study was to characterize extremely negative reviews of pediatric orthopaedic surgeons. Methods: Pediatric orthopaedic surgeons were randomly selected using the Pediatric Orthopaedic Society of North America comprehensive list of surgeons. A search was then performed on Healthgrades.com, Vitals.com, and Yelp.com for 1-star reviews. Reviews were classified into clinical and nonclinical categories. Statistical analyses were performed regarding the frequency of reviews and complaints for each respective category. Results: Of the 279 one-star reviews categorized, 248 reviews (88.9% of reviews) included nonclinical complaints, and 182 reviews (65.2% of reviews) included clinical complaints. Nonsurgical patients were associated with 255 reviews, and the remaining 24 were related to surgical patients. Of the 430 comments within reviews, 248 referenced nonclinical aspects of care, and 182 referenced clinical care. Clinical factors most frequently noted included clinical disagreement (37%), unclear treatment plan (25%), complication (17%), misdiagnosis (15%), uncontrolled pain (13%), and delay in care (8%). The most addressed nonclinical factors included physician bedside manner (68%), time spent with provider (21%), wait time (18%), unprofessional staff (17%), scheduling issues (9%), cost (8%), and billing (8%). Compared with surgical reviews, nonsurgical reviews were more likely to contain nonclinical complaints (rate ratio: 1.5; P < 0.05) and less likely to contain clinical complaints (rate ratio: 0.7; P < 0.05). The most common complaint by surgical patients was complications (91.7%). Conclusions: To our knowledge, this is the first study to examine the factors associated with negative reviews of pediatric orthopaedic surgeons. The majority of reviews of pediatric orthopaedic surgeons were left by nonsurgical patients and were related to nonclinical aspects of care. We also found surgeon-dependent factors such as poor physician bedside manner, unclear treatment plan, or parents' disagreement with treatment plan were the most common reasons for negative reviews. Level of Evidence: Level IV.