Gene therapy reforms photoreceptor structure and restores vision in NPHP5-associated Leber congenital amaurosis.

The inherited childhood blindness caused by mutations in NPHP5, a form of Leber congenital amaurosis, results in abnormal development, dysfunction, and degeneration of photoreceptors. A naturally occurring NPHP5 mutation in dogs leads to a phenotype that very nearly duplicates the human retinopathy in terms of the photoreceptors involved, spatial distribution of degeneration, and the natural history of vision loss. We show that adeno-associated virus (AAV)-mediated NPHP5 gene augmentation of mutant canine retinas at the time of active degeneration and peak cell death stably restores photoreceptor structure, function, and vision with either the canine or human NPHP5 transgenes. Mutant cone photoreceptors, which failed to form outer segments during development, reform this structure after treatment. Degenerating rod photoreceptor outer segments are stabilized and develop normal structure. This process begins within 8 weeks after treatment and remains stable throughout the 6-month posttreatment period. In both photoreceptor cell classes mislocalization of rod and cone opsins is minimized or reversed. Retinal function and functional vision are restored. Efficacy of gene therapy in this large animal ciliopathy model of Leber congenital amaurosis provides a path for translation to human treatment.

Restoration of Vision and Retinal Responses After Adeno-Associated Virus-Mediated Optogenetic Therapy in Blind Dogs.

Purpose: Optogenetic gene therapy to render remaining retinal cells light-sensitive in end-stage retinal degeneration is a promising strategy for treatment of individuals blind because of a variety of different inherited retinal degenerations. The clinical trials currently in progress focus on delivery of optogenetic genes to ganglion cells. Delivery of optogenetic molecules to cells in the outer neural retina is predicted to be even more advantageous because it harnesses more of the retinal circuitry. However, this approach has not yet been tested in large animal models. For this reason, we evaluated the safety and efficacy of optogenetic therapy targeting remaining diseased cone photoreceptors in the Rcd1 dog model of retinitis pigmentosa. Methods: Imaging and measures of retinal function and functional vision were carried out, as well as terminal studies evaluating multi-electrode array recordings and histology. Results: Animals remained healthy and active throughout the study and showed improved retinal and visual function as assessed by electroretinography and visual-evoked potentials, improved navigational vision, and improved function of cone photoreceptors and the downstream retinal circuitry. Conclusions: The findings demonstrate that an optogenetic approach targeting the outer retina in a blind large animal model can partially restore vision. Translational Relevance: This work has translational relevance because the approach could potentially be extrapolated to treat humans who are totally blind because of retinal degenerative disease.

Flow-mode water treatment under simultaneous hydrodynamic cavitation and plasma.

Over the last two decades, the scientific community and industry have made huge efforts to develop environmental protection technologies. In particular, the scarcity of drinking water has prompted the investigation of several physico-chemical treatments, and synergistic effects have been observed in hyphenated techniques. Herein, we report the first example of water treatment under simultaneous hydrodynamic cavitation and plasma discharge with the intense generation of radicals, UV light, shock waves and charged particles. This highly reactive environment is well suited to the bulk treatment of polluted water (i.e. E. coli disinfection and organic pollutant degradation). We have developed a new prototype and have efficiently applied this hybrid technology to water disinfection and the complete degradation of methanol in water with the aim of demonstrating its scalability. We have analyzed the mechanisms of water disinfection under the abovementioned conditions and verified them by measuring cavitation noise spectra and plasma emission spectra. We have also used the degradation of textile dyes and methanol solutions as an indicator for the formation of radicals.

Sonozonation (sonication/ozonation) for the degradation of organic contaminants - A review.

Ozonation (OZ) is an important advanced oxidation process to purify water and wastewater. Because of the lower solubility and instability of ozone (O3), selective oxidation and dependence on pH value, the industrial applications of OZ have been hindered by the following disadvantages: incomplete removal of pollutants, lower mineralization efficiency and the formation of toxic by-products. Meanwhile, OZ seems to have higher processing costs than other technologies. To improve the treatment efficiency and O3 utilization, several combined processes, such as H2O2/O3, UV/O3, and Cavitation/O3, have been explored, while the combined method of ultrasonication (US) with OZ is a promising treatment technology with a complex physicochemical mechanism. In US alone, the sonolysis of water molecules can produce more powerful unselective oxidant hydroxyl radicals (OH), and directly cause the sonochemical pyrolysis of volatile pollutants. In US/OZ, US can promote the mass transfer of O3, and also drive the chemical conversion of O3 to enhance the formation of OH. Various layouts of US/OZ devices and the interactive effects of US/OZ (synergism or antagonism) on the degradation of various organics are illustrated in this review. The main factors, including US frequency, pH value, and radical scavengers, significantly affect the mass transfer and decomposition of O3, the formation of OH and H2O2, the degradation rates of organics and the removal efficiencies of COD and TOC (mineralization). As a result, US can significantly increase the yield of OH, thereby improving the degradation efficiency and mineralization of refractory organics. However, US also enhances the decomposition of ozone, thereby reducing the concentration of O3 in water and impairing the efficiency of selective oxidation with O3 molecules.

The pecularities of ultrasonic equipment design for stabilization of dispersed structures of alumosilicic reagents for wastewater treatment.

Acoustic fields formed during operation of ultrasonic reactors with waveguides of following types: rod-type, cylindrical with rectangular protrusions and tubular were calculated and measured. The influence of distribution of acoustic fields arising from the operation of waveguide systems of three different types on the efficiency of ultrasonic activation of alumosilicic flocculant-coagulant and magnetite intended for water purification was investigated. It was shown that regardless of the equipment used on an industrial scale it is possible to reactivate the alumosilicic flocculant-coagulant even after the shelf life period of it passed, however in case of activation of magnetite the use of a bigger reactor in inefficient. In case of industrial scale processes, the choice of the correct reactor design is of significant importance, since it allows to reduce the required processing time, and, as a result, the energy consumption of the processes. The advantages of tubular waveguide systems include the possibility of processing large volumes of liquid. The high efficiency and uniformity of the excited ultrasonic fields can lead to reduction of operating costs. In case of smaller flows, the waveguide system with rectangular protrusions allowed to obtain better results. Our work illustrates the dependence of the success of a specific method on the choice of the waveguide and the size of the reactor during upscale.

Dual SMAD inhibition and Wnt inhibition enable efficient and reproducible differentiations of induced pluripotent stem cells into retinal ganglion cells.

Glaucoma is a group of progressive optic neuropathies that share common biological and clinical characteristics including irreversible changes to the optic nerve and visual field loss caused by the death of retinal ganglion cells (RGCs). The loss of RGCs manifests as characteristic cupping or optic nerve degeneration, resulting in visual field loss in patients with Glaucoma. Published studies on in vitro RGC differentiation from stem cells utilized classical RGC signaling pathways mimicking retinal development in vivo. Although many strategies allowed for the generation of RGCs, increased variability between experiments and lower yield hampered the cross comparison between individual lines and between experiments. To address this critical need, we developed a reproducible chemically defined in vitro methodology for generating retinal progenitor cell (RPC) populations from iPSCs, that are efficiently directed towards RGC lineage. Using this method, we reproducibly differentiated iPSCs into RGCs with greater than 80% purity, without any genetic modifications. We used small molecules and peptide modulators to inhibit BMP, TGF-ß (SMAD), and canonical Wnt pathways that reduced variability between iPSC lines and yielded functional and mature iPSC-RGCs. Using CD90.2 antibody and Magnetic Activated Cell Sorter (MACS) technique, we successfully purified Thy-1 positive RGCs with nearly 95% purity.

A method for water well regeneration based on shock waves and ultrasound.

The regeneration of water wells is an urgent problem nowadays, when drilling of new wells becomes more and more expensive. Formation damage leads to a reduction of the formation's permeability and/or pore volume which in turn inhibits the ability of the water to flow from the reservoir formation into the wellbore. A new technology that uses high-power ultrasound to remove formation damage of water wells has been developed. The effectiveness of regeneration of wells can be enhanced if ultrasound and shockwaves are used during the same treatment. It was shown by computer modelling, that the two methods have different depths of impact. Whereas the ultrasonic method has a strong impact on the area of the filter tube, the impact of the shock waves is focused on the gavel pack, the wall of the well and the adjacent aquifer. A shockwave treatment, which is normally more effective due to larger impact zone, needs to be followed by ultrasonic treatment in order to facilitate the removal of the detached deposits. These theoretical assumptions were confirmed by field tests on two wells. The use of the method leaded to an increase of the production by 40% and 109% respectively.