Optineurin mediates delivery of membrane from transferrin receptor-positive endosomes to autophagosomes

Autophagy is a conserved quality control mechanism that removes damaged proteins, organelles and invading bacteria through lysosome-mediated degradation. During autophagy several organelles including endoplasmic reticulum, mitochondria, plasma membrane and endosomes contribute membrane for autophagosome formation. However, the mechanisms and proteins involved in membrane delivery to autophagosomes are not clear. Optineurin (OPTN), a cytoplasmic adaptor protein, is involved in promoting maturation of phagophores into autophagosomes; it is also involved in regulating endocytic trafficking and recycling of transferrin receptor (TFRC). Here, we have examined the role of optineurin in the delivery of membrane from TFRC-positive endosomes to autophagosomes. Only a small fraction of autophagosomes was positive for TFRC, indicating that TFRC-positive endosomes could contribute membrane to a subset of autophagosomes. The percentage of TFRC-positive autophagosomes was reduced in Optineurin knockout mouse embryonic fibroblasts (Optn-/-MEFs) in comparison with normal MEFs. Upon over-expression of optineurin, the percentage of TFRC-positive autophagosomes was increased in Optn-/- MEFs. Unlike wild-type optineurin, a disease-associated mutant, E478G, defective in ubiquitin binding, was not able to enhance formation of TFRC-positive autophagosomes in Optn-/- MEFs. TFRC degradation mediated by autophagy was decreased in optineurin deficient cells. Our results suggest that optineurin mediates delivery of TFRC and perhaps associated membrane from TFRC-positive endosomes to autophagosomes, and this may contribute to autophagosome formation.

Influence of Adaptive Gap Control Mechanism and Tool Electrodes on Machining Titanium (Ti-6Al-4V) Alloy in EDM Process

Titanium alloy is widely used for orthodontic technology and easily machined using the EDM process. In the EDM process, the workpiece and tool electrode must be separated by a continuous air gap during the machining operation to generate discharge energy in this method. In the present study, an endeavor was made to analyze the effects of a servo feed air gap control and tool electrode in the EDM process. The developed mechanical setup consists of a linear action movement with zero backlash along the X-axis, which can be controlled up to 0.03 mm. It was observed that the suggested air gap control scheme can enhance the servo feed mechanism on a machining titanium alloy. A tungsten carbide electrode can enhance the surface measures owing to its ability to produce tiny craters with uniform distribution. Since it produces a little crater and has a higher melting point, a tungsten carbide electrode can create lesser surface roughness than a copper tool and brass tool electrode.

Analysis of the parameters of the double clutch drive with reduced control energy consumption

The global trend of improving vehicle and traction transmissions is associated with the use of robotic preselector gearboxes, an important component of which is the friction clutch. The review of existing designs and analysis of clutch development concepts allows determining the directions of their improvement, in particular, the improvement of dry double clutches, which is the subject of research in this paper. A significant disadvantage of existing dual-clutch designs is the use of additional special (mostly hydraulic or combined) systems for on-off friction pairs, so the structural cost and energy costs for control are increased, and the design, maintenance, and repair are complicated. The proposed design of the drive to control the original dry dual-clutch involves a lever mechanism with the use of rotary stops, which are made in the form of mobile carriages with rollers, this provides switching clutches in a short time and actually without interruption of power flow, so the acceleration dynamics of vehicles will be improved, the clutch design will be simplified and energy consumption for its control during the start and gear shifting will be reduced. The mathematical model of the proposed clutch drive on the basis of which the control mechanism operation is simulated and influence of its design parameters on operational indicators of the clutch drive, in particular, on kinematic characteristics of the drive, ranges of kinematic and power gear ratios, the power interaction of the mechanism links, the power of the actuator when each clutch is turning on. It is established that the switching of clutches requires less energy, the movement of the mechanism elements is different, and the compression force of the friction pairs when starting the first and second clutches is different. The simulation results confirm that the proposed original lever design of the clutch control mechanism is effective. The obtained results allow us to reasonably determine the parameters of a clutch, but the choice of optimal-rational parameters of its control mechanism requires further research, for which appropriate methods, search algorithms, and their software implementation have been developed.

Trans-Translation Is an Appealing Target for the Development of New Antimicrobial Compounds

Because of the ever-increasing multidrug resistance in microorganisms, it is crucial that we find and develop new antibiotics, especially molecules with different targets and mechanisms of action than those of the antibiotics in use today. Translation is a fundamental process that uses a large portion of the cell’s energy, and the ribosome is already the target of more than half of the antibiotics in clinical use. However, this process is highly regulated, and its quality control machinery is actively studied as a possible target for new inhibitors. In bacteria, ribosomal stalling is a frequent event that jeopardizes bacterial wellness, and the most severe form occurs when ribosomes stall at the 3′-end of mRNA molecules devoid of a stop codon. Trans-translation is the principal and most sophisticated quality control mechanism for solving this problem, which would otherwise result in inefficient or even toxic protein synthesis. It is based on the complex made by tmRNA and SmpB, and because trans-translation is absent in eukaryotes, but necessary for bacterial fitness or survival, it is an exciting and realistic target for new antibiotics. Here, we describe the current and future prospects for developing what we hope will be a novel generation of trans-translation inhibitors.