Rapid Electrical Stimulation Increased Cardiac Apoptosis Through Disturbance of Calcium Homeostasis and Mitochondrial Dysfunction in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

Background/Aims: Heart failure induced by tachycardia, the most common arrhythmia, is frequently observed in clinical practice. This study was designed to investigate the underlying mechanisms. Methods: Rapid electrical stimulation (RES) at a frequency of 3 Hz was applied on human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) for 7 days, with 8 h/day and 24 h/day set to represent short-term and long-term tachycardia, respectively. Age-matched hiPSC-CMs without electrical stimulation or with slow electrical stimulation (1 Hz) were set as no electrical stimulation (NES) control or low-frequency electrical stimulation (LES) control. Following stimulation, JC-1 staining flow cytometry analysis was performed to examine mitochondrial conditions. Apoptosis in hiPSC-CMs was evaluated using Hoechst staining and Annexin V/propidium iodide (AV/PI) staining flow cytometry analysis. Calcium transients and L-type calcium currents were recorded to evaluate calcium homeostasis. Western blotting and qPCR were performed to evaluate the protein and mRNA expression levels of apoptosis-related genes and calcium homeostasis-regulated genes. Results: Compared to the controls, hiPSC-CMs following RES presented mitochondrial dysfunction and an increased apoptotic percentage. Amplitudes of calcium transients and L-type calcium currents were significantly decreased in hiPSC-CMs with RES. Molecular analysis demonstrated upregulated expression of Caspase3 and increased Bax/Bcl-2 ratio. Genes related to calcium re-sequence were downregulated, while phosphorylated Ca2+/calmodulin-dependent protein kinase II (CaMKII) was significantly upregulated following RES. There was no significant difference between the NES control and LES control groups in these aspects. Inhibition of CaMKII with 1 µM KN93 partly reversed these adverse effects of RES. Conclusion: RES on hiPSC-CMs disturbed calcium homeostasis, which led to mitochondrial stress, promoted cell apoptosis and caused electrophysiological remodeling in a time-dependent manner. CaMKII played a central role in the damages induced by RES, pharmacological inhibition of CaMKII activity partly reversed the adverse effects of RES on both structural and electrophysiological properties of cells

Biochar-based fertilizer: Supercharging root membrane potential and biomass yield of rice

Biochar-based compound fertilizers (BCF) and amendments have proven to enhance crop yields and modify soil properties (pH, nutrients, organic matter, structure etc.) and are now in commercial production in China. While there is a good understanding of the changes in soil properties following biochar addition, the interactions within the rhizosphere remain largely unstudied, with benefits to yield observed beyond the changes in soil properties alone. We investigated the rhizosphere interactions following the addition of an activated wheat straw BCF at an application rates of 0.25% (g·g−1 soil), which could potentially explain the increase of plant biomass (by 67%), herbage N (by 40%) and P (by 46%) uptake in the rice plants grown in the BCF-treated soil, compared to the rice plants grown in the soil with conventional fertilizer alone. Examination of the roots revealed that micron and submicron-sized biochar were embedded in the plaque layer. BCF increased soil Eh by 85 mV and increased the potential difference between the rhizosphere soil and the root membrane by 65 mV. This increased potential difference lowered the free energy required for root nutrient accumulation, potentially explaining greater plant nutrient content and biomass. We also demonstrate an increased abundance of plant-growth promoting bacteria and fungi in the rhizosphere. We suggest that the redox properties of the biochar cause major changes in electron status of rhizosphere soils that drive the observed agronomic benefits

The Characterization of the Inhibitory Substances Produced by <i>Bacillus pumilus</i> LYMC-3 and the Optimization of Fermentation Conditions

Bacillus pumilus plays a very important role in the biological control of plant diseases. In this study, we conducted indoor biocontrol experiments using the LYMC-3 strain of B. pumilus to assess its efficacy against poplar canker and tea gray blight. Simultaneously, we explored the optimal fermentation conditions for the production of inhibitory substances against Phomopsis macrospora and Pestalotiopsis vesicolor. Subsequently, the inhibitory substances were extracted crudely and subjected to a preliminary investigation of their properties. The results of the indoor biocontrol experiments showed that the LYMC-3 strain exhibited disease prevention rates of 44.12% and 48.59% against poplar canker and tea gray blight, respectively. The fermentation optimization experiment showed that a culture medium containing 10 g/L of glucose, 15 g/L of tryptone, and 5 g/L of sodium chloride favored the production of inhibitory substances via the LYMC-3 strain. Among the crude extracts obtained, the non-protein extracts demonstrated the most obvious inhibitory effects. Furthermore, the inhibitory substances exhibited high polarity, excellent thermal stability, and high sensitivity to strong acids and bases. This study establishes a theoretical foundation for the purification and identification of inhibitory substances using the LYMC-3 strain. Additionally, it contributes to elucidating biological control mechanisms and facilitates the application of the LYMC-3 strain in production

An Inverse Kinematics Solution for a Series-Parallel Hybrid Banana-Harvesting Robot Based on Deep Reinforcement Learning

A series-parallel hybrid banana-harvesting robot was previously developed to pick bananas, with inverse kinematics intractable to an address. This paper investigates a deep reinforcement learning-based inverse kinematics solution to guide the banana-harvesting robot toward a specified target. Because deep reinforcement learning algorithms always struggle to explore huge robot workspaces, a practical technique called automatic goal generation is first developed. This draws random targets from a dynamic uniform distribution with increasing randomness to facilitate deep reinforcement learning algorithms to explore the entire robot workspace. Then, automatic goal generation is applied to a state-of-the-art deep reinforcement learning algorithm, the twin-delayed deep deterministic policy gradient, to learn an effective inverse kinematics solution. Simulation experiments show that with automatic goal generation, the twin-delayed deep deterministic policy gradient solved the inverse kinematics problem with a success rate of 96.1% and an average running time of 23.8 milliseconds; without automatic goal generation, the success rate was just 81.2%. Field experiments show that the proposed method successfully guided the robot to approach all targets. These demonstrate that automatic goal generation enables deep reinforcement learning to effectively explore the robot workspace and to learn a robust and efficient inverse kinematics policy, which can, therefore, be applied to the developed series-parallel hybrid banana-harvesting robot

Short-Term Trajectory Prediction of Maritime Vessel Using k-Nearest Neighbor Points

The prediction of ship location has become an increasingly popular research hotspot in the field of maritime transportation engineering, which benefits maritime safety supervision and security. Existing methods of ship location prediction based on motion characteristics have a large uncertainty and cannot guarantee trajectory prediction accuracy of the target ship. An improved method of location prediction using k-nearest neighbor (KNN) is proposed in this paper. An expanded circle area of the latest point of the target ship is first generated to find the reference points with similar movement characteristics in the constraints of distance and time intervals. Then, the top k-nearest neighbors are determined based on the degree of similarity. Relationships between the reference point of each neighbor and the latest points of the target ship are calculated. The predicted location of the target ship can then be determined by a weighted calculation of the locations of all neighbors at the predicted time and their relationships with the target ship. Experiments of ship location prediction in 10 min, 20 min, and 30 min were conducted. The correlation coefficient of the location prediction error for the three experiments was 0.992, 0.99, and 0.9875, respectively. The results show that ship location prediction with reference to multiple nearest neighbors with similar movements can provide better accuracy

Advances in bioleaching of waste lithium batteries under metal ion stress

In modern societies, the accumulation of vast amounts of waste Li-ion batteries (WLIBs) is a grave concern. Bioleaching has great potential for the economic recovery of valuable metals from various electronic wastes. It has been successfully applied in mining on commercial scales. Bioleaching of WLIBs can not only recover valuable metals but also prevent environmental pollution. Many acidophilic microorganisms (APM) have been used in bioleaching of natural ores and urban mines. However, the activities of the growth and metabolism of APM are seriously inhibited by the high concentrations of heavy metal ions released by the bio-solubilization process, which slows down bioleaching over time. Only when the response mechanism of APM to harsh conditions is well understood, effective strategies to address this critical operational hurdle can be obtained. In this review, a multi-scale approach is used to summarize studies on the characteristics of bioleaching processes under metal ion stress. The response mechanisms of bacteria, including the mRNA expression levels of intracellular genes related to heavy metal ion resistance, are also reviewed. Alleviation of metal ion stress via addition of chemicals, such as spermine and glutathione is discussed. Monitoring using electrochemical characteristics of APM biofilms under metal ion stress is explored. In conclusion, effective engineering strategies can be proposed based on a deep understanding of the response mechanisms of APM to metal ion stress, which have been used to improve bioleaching efficiency effectively in lab tests. It is very important to engineer new bioleaching strains with high resistance to metal ions using gene editing and synthetic biotechnology in the near future

Oxidative Stress Induced by Metal Ions in Bioleaching of LiCoO2 by an Acidophilic Microbial Consortium

© Copyright © 2020 Liu, Liu, Wu, Zhang, Gu, Zhu and Tan. An acidophilic microbial consortium (AMC) was used to investigate the fundamental mechanism behind the adverse effects of pulp density increase in the bioleaching of waste lithium ion batteries (WLIBs). Results showed that there existed the effect of metal-ion stress on the bio-oxidative activity of AMC. The Li+ and Co2+ accumulated in the leachate were the direct cause for the decrease in lithium and cobalt recovery yields under a high pulp density. In a simulated bioleaching system with 4.0% (w ⋅v–1) LiCoO2, the intracellular reactive oxygen species (ROS) content in AMC increased from 0.82 to 6.02 within 24 h, which was almost three times higher than that of the control (2.04). After the supplementation of 0.30 g⋅L–1 of exogenous glutathione (GSH), the bacterial intracellular ROS content decreased by 40% within 24 h and the activities of intracellular ROS scavenging enzymes, including glutathione peroxidase (GSH-Px) and catalase (CAT), were 1.4- and 2.0-folds higher in comparison with the control within 24 h. In the biofilms formed on pyrite in the bioleaching of WLIBs, it was found that metal-ion stress had a great influence on the 3-D structure and the amount of biomass of the biofilms. After the exogenous addition of GSH, the structure and the amount of biomass of the biofilms were restored to some extent. Eventually, through ROS regulation by the exogenous addition of GSH, very high metal recovery yields of 98.1% Li and 96.3% Co were obtained at 5.0% pulp density