No abstract

Phosphatase and tensin homolog (PTEN)-induced putative kinase 1 (PINK1) is a gatekeeper of mitochondrial quality control. The present study was aimed to examine whether PINK1 possesses a protective function against gentamicin (GM)-induced sensory hair cell (HC) damage in vitro. The formation of parkin particles (a marker revealing the activation of PINK1 pathway which is a substrate of PINK1 and could signal depolarized mitochondria for clearance) and autophagy were determined by immunofluorescence staining. The expressions of PINK1, LC3B, cleaved-caspase 3 and p53 were measured by Western blotting. The levels of reactive oxygen species (ROS) and apoptosis were respectively evaluated by DCFH-DA staining, Annexin V Apoptosis Detection Kit and TUNEL staining. Cell viability was tested by a CCK8 kit. We found that treatment of 400 μM GM elicited the formation of ROS, which, in turn, led to PINK1 degradation, parkin recruitment, autophagy formation, an increase of p53 and cleaved-caspase 3 in HEI-OC1 cells and murine HCs. In contrast, co-treatment with ROS scavenger N-acetyl-L-cysteine (NAC) inhibited parkin recruitment, alleviated autophagy and p53 pathway-related damaged-cell elimination. Moreover, PINK1 interference contributed to a decrease of autophagy but an increase of p53 level in HEI-OC1 cells in response to GM stimulus. Findings from this work indicate that PINK1 alleviates the GM-elicited ototoxicity via induction of autophagy and resistance the increase of p53 in HCs.

The objective of this study was to elucidate whether paeoniflorin (PF) exerted an effect on cisplatin‐induced spiral ganglion neuron (SGN) damage, with special attention given to the role of PINK1/BAD pathway in this process. Middle cochlear turn culture and C57BL/6 mice were utilized to identify the character of PF in vitro and in vivo. We found that cisplatin treatment led to SGN damage, in which reactive oxygen species (ROS) generation increased, PINK1 expression decreased, BAD accumulation on mitochondria raised and mitochondrial apoptotic pathway activated. Conversely, we demonstrated that PF pre‐treatment obviously mitigated cisplatin‐induced SGN damage. Mechanistic studies showed that PF could reduce ROS levels, increase PINK1 expression, decrease the BAD accumulation on mitochondria and, thus, alleviate the activated mitochondrial apoptosis in SGNs caused by cisplatin. Overall, the findings from this work reveal the important role of PF and provide another strategy against cisplatin‐induced ototoxicity.

The autonomous shuttle and stacker crane (AC/SC) warehousing system, as a new automated deep-lane unit load storage/retrieval system, has been becoming more popular, especially for batch order fulfilment because of its high flexibility, low operational cost and improved storage capacity. This system consists of a shuttle sub-system that controls motion along the x-axis and a stacker crane sub-system that controls motion along the y-axis and z-axis. The combination of shuttles and a stacker crane performs storage and retrieval tasks. Modelling the parallel motion is an important design tool that can be used to calculate the optimal number of shuttles for a given configuration of the warehousing system. In this study, shuttle movements from one lane to another are inserted into the stock-keeping unit (SKU) task queue, and convert such that they are consistent with the retrieval tasks. The tasks are then grouped according to their starting lane, and converted to an assembly-line parallel job problem by analysing the operating mode with the objectives of minimising the total working time of the stacker crane and the wasted shuttle time. A time sequence mathematical model based on the motion of the shuttles and stacker crane is proposed, and an improved Pareto-optimal elitist non-dominated sorting genetic algorithm is used to solve this multi-objective optimization problem. The model is validated via a simulation study, and via a real-world warehousing case study. We go on to describe guidelines for the layout and configuration of AS/SC warehousing systems, including the optimal number of shuttles and number of x-axis storage cells of lanes, which can improve efficiency and minimise both capital investment and operating costs.