Primary care physician and resident perceptions of gun safety counseling.

A significant public health burden exists related to firearm-related injuries in the United States. Primary care physicians (PCPs) find themselves on the frontline to address it. This study aimed to determine if PCPs feel prepared and responsible for providing firearm safety counseling. Data were gathered via a direct 23-question survey of family medicine, internal medicine, and pediatric physicians and residents in North and Central Texas between May 2019 and January 2020. The survey was delivered to 518 outpatient providers, with a 24% response rate. Associations were assessed by chi-square/Fisher's exact test. A total of 62% agreed that preventable firearm death is an important health issue and PCPs can have a positive effect; 10% felt their residency provided or provides opportunities to learn about firearm safety and how to counsel patients; 55% agreed that a standardized tool to question and counsel patients on firearm safety such as a template in the electronic health record would be useful; and 68% stated they would use a standardized template in their practice if proven to reduce firearm-related injuries. Physicians who had received training in residency felt their knowledge was adequate to be able to provide meaningful counseling to patients (P < 0.001). In conclusion, PCPs are pivotal in addressing firearm safety. A formal curriculum across primary care specialties' graduate education programs should be developed to improve physicians' competency in addressing firearm safety. Further investigation into standardized tools that could be implemented into an electronic health record may help bridge the gap from awareness to implementation of interventions.

Synthetic Adeno-Associated Viral Vector Efficiently Targets Mouse and Nonhuman Primate Retina In Vivo.

Gene therapy is a promising approach in the treatment of inherited and common complex disorders of the retina. Preclinical and clinical studies have validated the use of adeno-associated viral vectors (AAV) as a safe and efficient delivery vehicle for gene transfer. Retinal pigment epithelium and rods-and to a lesser extent, cone photoreceptors-can be efficiently targeted with AAV. Other retinal cell types however are more challenging targets. The aim of this study was to characterize the transduction profile and efficiency of in silico designed, synthetic Anc80 AAVs for retinal gene transfer. Three Anc80 variants were evaluated for retinal targeting in mice and primates following subretinal delivery. In the murine retina Anc80L65 demonstrated high level of retinal pigment epithelium and photoreceptor targeting with comparable cone photoreceptor affinity compared to other AAVs. Remarkably, Anc80L65 enhanced transduction kinetics with visible expression as early as day 1 and steady state mRNA levels at day 3. Inner retinal tropism of Anc80 variants demonstrated distinct transduction patterns of Müller glia, retinal ganglion cells and inner nuclear layer neurons. Finally, murine findings with Anc80L65 qualitatively translated to the Rhesus macaque in terms of cell targets, levels and onset of expression. Our findings support the use of Anc80L65 for therapeutic subretinal gene delivery.

In Silico Reconstruction of the Viral Evolutionary Lineage Yields a Potent Gene Therapy Vector.

Adeno-associated virus (AAV) vectors have emerged as a gene-delivery platform with demonstrated safety and efficacy in a handful of clinical trials for monogenic disorders. However, limitations of the current generation vectors often prevent broader application of AAV gene therapy. Efforts to engineer AAV vectors have been hampered by a limited understanding of the structure-function relationship of the complex multimeric icosahedral architecture of the particle. To develop additional reagents pertinent to further our insight into AAVs, we inferred evolutionary intermediates of the viral capsid using ancestral sequence reconstruction. In-silico-derived sequences were synthesized de novo and characterized for biological properties relevant to clinical applications. This effort led to the generation of nine functional putative ancestral AAVs and the identification of Anc80, the predicted ancestor of the widely studied AAV serotypes 1, 2, 8, and 9, as a highly potent in vivo gene therapy vector for targeting liver, muscle, and retina.