Heparan Sulfate Binding Promotes Accumulation of Intravitreally Delivered Adeno-associated Viral Vectors at the Retina for Enhanced Transduction but Weakly Influences Tropism

ABSTRACT Many adeno-associated virus (AAV) serotypes efficiently transduce the retina when delivered to the subretinal space but show limited success when delivered to the vitreous due to the inner limiting membrane (ILM). Subretinal delivery of AAV serotype 2 (AAV2) and its heparan sulfate (HS)-binding-deficient capsid led to similar expression, indicating transduction of the outer retina occurred by HS-independent mechanisms. However, intravitreal delivery of HS-ablated recombinant AAV2 (rAAV2) led to a 300-fold decrease in transduction compared to AAV2. Fluorescence in situ hybridization of AAV transgenes was used to identify differences in retinal trafficking and revealed that HS binding was responsible for AAV2 accumulation at the ILM. This mechanism was tested on human ex vivo retinas and showed similar accumulation with HS-binding AAV2 capsid only. To evaluate if HS binding could be applied to other AAV serotypes to enhance their transduction, AAV1 and AAV8 were modified to bind HS with a single-amino-acid mutation and tested in mice. Both HS-binding mutants of AAV1 and AAV8 had higher intravitreal transduction than their non-HS-binding parent capsid due to increased retinal accumulation. To understand the influence that HS binding has on tropism, chimeric AAV2 capsids with dual-glycan usage were tested intravitreally in mice. Compared to HS binding alone, these chimeric capsids displayed enhanced transduction that was correlated with a change in tropism. Taken together, these data indicate that HS binding serves to sequester AAV capsids from the vitreous to the ILM but does not influence retinal tropism. The enhanced retinal transduction of HS-binding capsids provides a rational design strategy for engineering capsids for intravitreal delivery. IMPORTANCE Adeno-associated virus (AAV) has become the vector of choice for viral gene transfer and has shown great promise in clinical trials. The need for development of an easy, less invasive injection route for ocular gene therapy is met by intravitreal delivery, but delivery of AAV by this route results in poor transduction outcomes. The inner limiting membrane (ILM) creates a barrier separating the vitreous and the retina. Binding of AAV to heparan sulfate proteoglycan (HSPG) at the ILM may allow the virus to traverse this barrier for better retinal transduction. We show that HSPG binding is correlated with greater accumulation and penetration of AAV in the retina. We demonstrated that this accumulation is conserved across mouse and human retinas and that the addition of HSPG binding to other AAV capsids can increase the number of vectors accumulating at the ILM without dictating tropism

Young Hispanics’ Motivations to Use Smartphones: A Three-Country Comparative Study

The fast-changing digital media environment has provoked new communication processes in contemporary society. In particular, younger generations engage in new ways of interacting with each other thanks to the constant emergence of digital technologies and mobile communication outlets. New habits arise every year, changing not only our interpersonal communication, but also our overall consumption of information and entertainment content, with video and instant messaging apps being particularly popular among younger populations. This study employs a mixed method approach to measure young Hispanics’ motivations to use smartphones in Chile, Mexico and the U.S. The subjects, 18 to 25 years old, participated in focus groups and surveys conducted in 2015: 55 Hispanic smartphone users joined focus groups whilst 1,403 respondents completed surveys in Chile, Mexico and Texas (U.S.). Results show similarities and differences in young Hispanics’ motivations and uses of smartphones, including the personal and commercial value they ascribe to information, time spent on social networks, and their preferences for mobile devices compared to other media. The findings are used to develop three archetypes of young users, based on smartphone’s motivations and uses. Utilizing factor analysis scores, a linear regression analysis was conducted to identify the main traits of these three archetypes: “always connected,” “entertained,” and “secure.

A high resolution study of dynamic changes of Ce2O3 and CeO2 nanoparticles in complex environmental media

Ceria nanoparticles (NPs) rapidly and easily cycle between Ce(III) and Ce(IV) oxidation states, making them prime candidates for commercial and other applications. Increased commercial use has resulted in increased discharge to the environment and increased associated risk. Once in complex media such as environmental waters or toxicology exposure media, the same redox transformations can occur, causing altered behavior and effects compared to the pristine NPs. This study used high resolution scanning transmission electron microscopy and electron energy loss spectroscopy to investigate changes in structure and oxidation state of small, polymer-coated ceria suspensions in complex media. NPs initially in either the III or IV oxidation states, but otherwise identical, were used. Ce(IV) NPs were changed to mixed (III, IV) NPs at high ionic strengths, while the presence of natural organic macromolecules (NOM) stabilized the oxidation state and increased crystallinity. The Ce(III) NPs remained as Ce(III) at high ionic strengths, but were modified by the presence of NOM, causing reduced crystallinity and degradation of the NPs. Subtle changes to NP properties upon addition to environmental or ecotoxicology media suggest that there may be small but important effects on fate and effects of NPs compared to their pristine form

Dealing with mobility: Understanding access anytime, anywhere

The rapid and accelerating move towards the adoption and use of mobile technologies has increasingly provided people and organisations with the ability to work away from the office and on the move. The new ways of working afforded by these technologies are often characterised in terms of access to information and people ‘anytime, anywhere’. This paper presents a study of mobile workers that highlights different facets of access to remote people and information, and different facets of anytime, anywhere. Four key factors in mobile work are identified from the study: the role of planning, working in ‘dead time’, accessing remote technological and informational resources, and monitoring the activities of remote colleagues. By reflecting on these issues, we can better understand the role of technology and artefact use in mobile work and identify the opportunities for the development of appropriate technological solutions to support mobile workers

Transplanckian axions !?

We discuss quantum gravitational effects in Einstein theory coupled to periodic axion scalars to analyze the viability of several proposals to achieve superplanckian axion periods (aka decay constants) and their possible application to large field inflation models. The effects we study correspond to the nucleation of euclidean gravitational instantons charged under the axion, and our results are essentially compatible with (but independent of) the Weak Gravity Conjecture, as follows: Single axion theories with superplanckian periods contain gravitational instantons inducing sizable higher harmonics in the axion potential, which spoil superplanckian inflaton field range. A similar result holds for multi-axion models with lattice alignment (like the Kim-Nilles-Peloso model). Finally, theories with $N$ axions can still achieve a moderately superplanckian periodicity (by a $\sqrt{N}$ factor) with no higher harmonics in the axion potential. The Weak Gravity Conjecture fails to hold in this case due to the absence of some instantons, which are forbidden by a discrete $\mathbf{Z}_N$ gauge symmetry. Finally we discuss the realization of these instantons as euclidean D-branes in string compactifications.Comment: 46 pages, 6 figures. Added references, clarifications, and missing factor of 1/2 to instanton action. Conclusions unchange

Characterization of Fast Ion Transport via Position-Dependent Optical Deshelving

Ion transport is an essential operation in some models of quantum information processing, where fast ion shuttling with minimal motional excitation is necessary for efficient, high-fidelity quantum logic. While fast and cold ion shuttling has been demonstrated, the dynamics and specific trajectory of an ion during diabatic transport have not been studied in detail. Here we describe a position-dependent optical deshelving technique useful for sampling an ion's position throughout its trajectory, and we demonstrate the technique on fast linear transport of a $^{40}\text{Ca}^+$ ion in a surface-electrode ion trap. At high speed, the trap's electrode filters strongly distort the transport potential waveform. With this technique, we observe deviations from the intended constant-velocity (100 m/s) transport: we measure an average speed of 83(2) m/s and a peak speed of 251(6) m/s over a distance of 120 $\mu$