•Strong acidity removes the economic hurdle of fluidized struvite crystallization.•A 12 m3/d pilot-scale system is built to evaluate the performance on-site.•P recovery rate is achieved at 99.2% via ...internal circulation mode.•Using MgO with MgCl2 supplement reduces 42.8% of the reagent cost.•Struvite technology saves up to 31.33 million CYN over 20 years at 500 m3/d scale.
Production of wood-based activated carbon (WAC) generates large volume of highly acidic and phosphate-rich wastewater. Currently, the routine treatment (i.e. lime precipitation) creates significant secondary pollution, leading to extra economic and environmental burdens. Here, by exploiting the strong acidity of WAC wastewater, we successfully demonstrate fluidized struvite crystallization as a low-cost treatment alternative. Based on a 12 m3/d on-site pilot-scale system, four different fluidized struvite crystallization scenarios are evaluated from technical, economic, and environmental perspectives. The results show that using MgO with MgCl2 supplement saves 42.8% of the reagent cost when treating phosphate-rich wastewater (i.e. P = 3125.2 mg/L), and also maintains ideal P removal rate and struvite product purity. Meanwhile, the internal circulation mode exhibits higher P recovery (99.2%) than the external mode (55.3%-89.3%), while also demonstrates superior economic and environmental benefit because of less chemical consumption. In addition, the struvite morphology can be turned between pellets with strong crushing strength (external mode) to powder (internal mode). By Life cycle cost (LCC) analysis, we find that, on a treatment scale of 500 m3/d, struvite-based technology saves up to 31.33 million Chinese Yuan (CYN) during a 20-year lifespan, with relative payback period of 2.60 year. The technical, economic, and environmental assessments confirm that the struvite technology is a promising alternative in solving the bottleneck of WAC wastewater treatment.
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Usher syndrome (USH) is the leading genetic cause of combined hearing and vision loss. Among the three USH clinical types, type 2 (USH2) occurs most commonly. USH2A, GPR98, and WHRN are three known ...causative genes of USH2, whereas PDZD7 is a modifier gene found in USH2 patients. The proteins encoded by these four USH genes have been proposed to form a multiprotein complex, the USH2 complex, due to interactions found among some of these proteins in vitro, their colocalization in vivo, and mutual dependence of some of these proteins for their normal in vivo localizations. However, evidence showing the formation of the USH2 complex is missing, and details on how this complex is formed remain elusive. Here, we systematically investigated interactions among the intracellular regions of the four USH proteins using colocalization, yeast two-hybrid, and pull-down assays. We show that multiple domains of the four USH proteins interact among one another. Importantly, both WHRN and PDZD7 are required for the complex formation with USH2A and GPR98. In this USH2 quaternary complex, WHRN prefers to bind to USH2A, whereas PDZD7 prefers to bind to GPR98. Interaction between WHRN and PDZD7 is the bridge between USH2A and GPR98. Additionally, the USH2 quaternary complex has a variable stoichiometry. These findings suggest that a non-obligate, short term, and dynamic USH2 quaternary protein complex may exist in vivo. Our work provides valuable insight into the physiological role of the USH2 complex in vivo and informs possible reconstruction of the USH2 complex for future therapy.Assembly of the protein complex associated with Usher syndrome type 2 (USH2) is unclear.
WHRN and PDZD7 heterodimerization and their interactions with USH2A and GPR98 are required for USH2 complex formation.
An USH2 quaternary protein complex may exist in vivo.
This study provides clues to USH2 pathogenesis and may permit the complex reconstitution for therapeutic intervention.
Whirlin mutations cause retinal degeneration and hearing loss in Usher syndrome type II (USH2) and non-syndromic deafness, DFNB31. Its protein recruits other USH2 causative proteins to form a complex ...at the periciliary membrane complex in photoreceptors and the ankle link of the stereocilia in hair cells. However, the biological function of this USH2 protein complex is largely unknown. Using a yeast two-hybrid screen, we identified espin, an actin-binding/bundling protein involved in human deafness when defective, as a whirlin-interacting protein. The interaction between these two proteins was confirmed by their coimmunoprecipitation and colocalization in cultured cells. This interaction involves multiple domains of both proteins and only occurs when espin does not bind to actin. Espin was partially colocalized with whirlin in the retina and the inner ear. In whirlin knockout mice, espin expression changed significantly in these two tissues. Further studies found that whirlin increased the mobility of espin and actin at the actin bundles cross-linked by espin and, eventually, affected the dimension of these actin bundles. In whirlin knockout mice, the stereocilia were thickened in inner hair cells. We conclude that the interaction between whirlin and espin and the balance between their expressions are required to maintain the actin bundle network in photoreceptors and hair cells. Disruption of this actin bundle network contributes to the pathogenic mechanism of hearing loss and retinal degeneration caused by whirlin and espin mutations. Espin is a component of the USH2 protein complex and could be a candidate gene for Usher syndrome.
Whirlin is the causative gene for Usher syndrome type IID (USH2D), a condition manifested as both retinitis pigmentosa and congenital deafness. Mutations in this gene cause disruption of the USH2 ...protein complex composed of USH2A and VLGR1 at the periciliary membrane complex (PMC) in photoreceptors. In this study, the adeno-associated virus (AAV)-mediated whirlin replacement was evaluated as a treatment option.
Murine whirlin cDNA driven by the human rhodopsin kinase promoter (hRK) was packaged as an AAV2/5 vector and delivered into the whirlin knockout retina through subretinal injection. The efficiency, efficacy, and safety of this treatment were examined using immunofluorescent staining, confocal imaging, immunoelectron microscopy, Western blot analysis, histologic analysis, and electroretinogram.
The AAV-mediated whirlin expression started at two weeks, reached its maximum level at 10 weeks, and lasted up to six months post injection. The transgenic whirlin product had a molecular size and an expression level comparable to the wild-type. It was distributed at the PMC in both rod and cone photoreceptors from the central to peripheral retina. Importantly, the transgenic whirlin restored the cellular localization and expression level of both USH2A and VLGR1 and did not cause defects in the retinal histology and function in the whirlin knockout mouse.
Whirlin transgene recruits USH2A and VLGR1 to the PMC and is sufficient for the formation of the USH2 protein complex in photoreceptors. The combined hRK and AAV gene delivery system could be an effective gene therapy approach to treat retinal degeneration in USH2D patients.
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