Wilson's disease (WD) is a monogenetic liver disease that is based on a mutation of the ATP7B gene and leads to a functional deterioration in copper (Cu) excretion in the liver. The excess Cu accumulates in various organs such as the liver and brain. WD patients show clinical heterogeneity, which can range from acute or chronic liver failure to neurological symptoms. The course of the disease can be improved by a lifelong treatment with zinc or chelators such as D-penicillamine in a majority of patients, but serious side effects have been observed in a significant portion of patients, e.g. neurological deterioration and nephrotoxicity, so that a liver transplant would be inevitable. An alternative therapy option would be the genetic correction of the ATP7B gene. The novel gene therapy method CRISPR/Cas9, which has recently been used in the clinic, may represent a suitable therapeutic opportunity. In this study, we first initiated an artificial ATP7B point mutation in a human cell line using CRISPR/Cas9 gene editing, and corrected this mutation by the additional use of singlestranded oligo DNA nucleotides (ssODNs), simulating a gene correction of a WD point mutation in vitro. By the addition of 0.5 mM of Cu three days after lipofection, a high yield of CRISPR/Cas9-mediated ATP7B repaired cell clones was achieved (60%). Moreover, the repair efficiency was enhanced using ssODNs that incorporated three blocking mutations. The repaired cell clones showed a high resistance to Cu after exposure to increasing Cu concentrations. Our findings indicate that CRISPR/Cas9-mediated correction of ATP7B point mutations is feasible and may have the potential to be transferred to the clinic.

Upcoming Energy related Products (ErP) regulations on wastewater pumps by the European Commission will affect all pump manufacturers and operators of wastewater systems. Hence, the preparation of efficiency standards for wastewater pumps is intensively accompanied by input from the affected stakeholders and experts of different fields [1]. The previous approaches of ErP regulations, as in lot 11 (Electric motors, Ventilation fans, Circulators in buildings, Electric pumps), focus only on efficiency. However, when applying the philosophy of Ecodesign directly to wastewater pumps, the complex flow structure and the transport behaviour of this inhomogeneous multi-phase fluid must be taken into account. While efficiency is an important criterion, it is necessary to take the specifics of sewage transport into account when designing a new test standard, so as not to compromise on proven and “system-efficient” technologies. Therefore, the Berlin Institute of Technology is currently investigating wastewater compositions and limits for reliable pump operation in order to design a test standard for wastewater pumps comparable to the DIN EN ISO 9906 efficiency tests for clear water [2]. The test will assess the functional fulfilment level of the pump performance, differentiating between the wastewater classes.

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To date, there is no standardised test available, which assesses the actual performance of a pump for wastewater (especially its clog-resistance) as well as providing references about its efficiency. The developed testing procedure presented in this paper can display the different levels of performance for different pumps. The functionality of the pump, together with its efficiency is aggregated to the Functional Efficiency Index (FEI). The developed testing procedure is validated with two pumps from the field. Both are considered to be very prone to clogging by their respective wastewater network operators.