Switch-2 determines Mg2+ADP-release kinetics and fine-tunes the duty ratio of Dictyostelium class-1 myosins.

Though myosins share a structurally conserved motor domain, single amino acid variations of active site elements, including the P-loop, switch-1 and switch-2, which act as nucleotide sensors, can substantially determine the kinetic signature of a myosin, i.e., to either perform fast movement or enable long-range transport and tension generation. Switch-2 essentially contributes to the ATP hydrolysis reaction and determines product release. With few exceptions, class-1 myosin harbor a tyrosine in the switch-2 consensus sequence DIYGFE, at a position where class-2 myosins and a selection of myosins from other classes have a substitution. Here, we addressed the role of the tyrosine in switch-2 of class-1 myosins as potential determinant of the duty ratio. We generated constitutively active motor domain constructs of two class-1 myosins from the social amoeba Dictyostelium discoideum, namely, Myo1E, a high duty ratio myosin and Myo1B, a low duty ratio myosin. In Myo1E we introduced mutation Y388F and in Myo1B mutation F387Y. The detailed functional characterization by steady-state and transient kinetic experiments, combined with in vitro motility and landing assays revealed an almost reciprocal relationship of a number of critical kinetic parameters and equilibrium constants between wild-type and mutants that dictate the lifetime of the strongly actin-attached states of myosin. The Y-to-F mutation increased the duty ratio of Moy1B by almost one order of magnitude, while the introduction of the phenylalanine in switch-2 of Myo1E transformed the myosin into a low duty ratio motor. These data together with structural considerations propose a role of switch-2 in fine-tuning ADP release through a mechanism, where the class-specific tyrosine together with surrounding residues contributes to the coordination of Mg2+ and ADP. Our results highlight the importance of conserved switch-2 residues in class-1 myosins for efficient chemo-mechanical coupling, revealing that switch-2 is important to adjust the duty ratio of the amoeboid class-1 myosins for performing movement, transport or gating functions.

Photovoltaic connected active switched boost quasi-Z-source (ASB-qZSI)-based multi-level inverter system using Q2OGEO approach.

An effective hybrid technique is proposed for enhancing the efficiency of photovoltaic (PV) system by an extended boost inverter called active-switched boost quasi-Z source inverter (ASB-qZSI). The hybrid approach is the combination of quasi-oppositional chemical reaction optimization (QOCRO) and golden eagle optimizer (GEO), and later called Q2OGEO approach. The main objective of the proposed system is "to increase the proficiency of PV system and the extraction of maximum power." Here, ASB-qZSI method is used to increase the voltage gain of converter in less duty cycles. The objective of the system is defined as controller-based parameters and limitations like voltages, modulation index, current, and power. The QOCRO method is utilized to regulate the system voltage, and GEO is used to regulate the current of system. The proposed Q2OGEO method efficiently controls the voltage, current, and DC link voltage and then extracts maximal power of PV system. Additionally, increasing the power delivery, voltage profile, but minimizing the total harmonic distortion (THD). By using the ASB-qZSI, control the shoot-through duty ratio to minimize the modulation burden. Then, the proposed approach is enthused on the MATLAB software, and the effectiveness is observed with existing techniques.

Duty ratio drive prediction model of lifetime degradation for organic light emitting diode-on-silicon microdisplay.

A duty ratio drive prediction (DRDP) model of luminance degradation for organic light emitting diodes (OLED) microdisplay is proposed in this paper. The traditional stretched exponential decay (SED) model is not applicable for OLED driven by duty ratio. The DRDP model introduces the duty ratio as the variables affecting the lifetime of OLED. By fitting the undetermined coefficients with the measured luminance data, the quantitative relationships among the initial luminance, duty ratio, and OLED lifetime are obtained. Meanwhile, the model quantifies the phenomenon of spontaneous luminance recovery, which occurs when OLED switches from bright to dark. Finally, the DRDP model is used to compensate the luminance degradation of OLED driven by duty ratio. The experimental results show that the average prediction accuracy of DRDP model for white, red, green, and blue (W/R/G/B) OLED degradation trend is 0.9623. The average prediction accuracy of W/R/G/B OLED lifetime is 0.6119, which is greater than that of SED model. The lifetime is extended by 89.83% after compensation.

Effect of Plasma On-Time with a Fixed Duty Ratio on Reactive Species in Plasma-Treated Medium and Its Significance in Biological Applications.

Optimizing the therapeutic range of nonthermal atmospheric pressure plasma (NTAPP) for biomedical applications is an active research topic. For the first time, we examined the effect of plasma on-times in this study while keeping the duty ratio and treatment time fixed. We have evaluated the electrical, optical, and soft jet properties for two different duty ratios of 10% and 36%, using the plasma on-times of 25, 50, 75, and 100 ms. Furthermore, the influence of plasma on-time on reactive oxygen and nitrogen species (ROS/RNS) levels in plasma treated medium (PTM) was also investigated. Following treatment, the characteristics of (DMEM media) and PTM (pH, EC, and ORP) were also examined. While EC and ORP rose by raising plasma on-time, pH remained unchanged. Finally, the PTM was used to observe the cell viability and ATP levels in U87-MG brain cancer cells. We found it interesting that, by increasing the plasma on-time, the levels of ROS/RNS dramatically increased in PTM and significantly affected the viability and ATP levels of the U87-MG cell line. The results of this study provide a significant indication of advancement by introducing the optimization of plasma on-time to increase the efficacy of the soft plasma jet for biomedical applications.

Influence of Cathode Channel Parameters and Fan Duty Ratio on Low Power Forced-Convection Open-Cathode Proton Exchange Membrane Fuel Cell Stack.

The open-cathode forced-convection proton exchange membrane fuel cell has emerged as a viable option for portable energy sources. The forced-convection open-cathode mode, however, makes the cell's performance sensitive to changes in the cathode channel and fan parameters. In this study, small fuel cell stacks with varying cathode channel depths, widths, and width-rib ratios were assembled, and the effects of different cathode channel parameters and fan duty ratios on cell performance were investigated. The experimental results show that changing the cathode channel parameters has a significant impact on oxidant supply. When the channel width is increased, the cell performance increases first, then decreases. The cell performance decreases as the channel width-rib ratio increases. The performance of the cell improves as the cathode channel depth increases. Furthermore, the experimental results show that decreasing the duty ratio of the fan and using moderate heating improves cell performance.

Efficient Illumination for a Light-Addressable Potentiometric Sensor.

A light-addressable potentiometric sensor (LAPS) is a chemical sensor that is based on the field effect in an electrolyte-insulator-semiconductor structure. It requires modulated illumination for generating an AC photocurrent signal that responds to the activity of target ions on the sensor surface. Although high-power illumination generates a large signal, which is advantageous in terms of the signal-to-noise ratio, excess light power can also be harmful to the sample and the measurement. In this study, we tested different waveforms of modulated illuminations to find an efficient illumination for a LAPS that can enlarge the signal as much as possible for the same input light power. The results showed that a square wave with a low duty ratio was more efficient than a sine wave by a factor of about two.

Research on Multi-Physics Coupling Simulation for the Pulse Electrochemical Machining of Holes with Tube Electrodes.

Electrical parameters of the power supply are significant factors affecting the accuracy and stability of the electrochemical machining (ECM). However, the electric field, flow velocity and temperature in the machining area are difficult to measure directly under the influence of the power supply. Therefore, taking the film cooling hole as an example, the multi-physics coupling simulation analysis of the ECM is performed on the basis of Faraday's law and fluid heat transfer mathematical model. The machining characteristics of the direct current and pulse ECM are compared through simulation. The results show that the pulse ECM improves the distribution of temperature and current density in the machining area. The period has little effect on the temperature, current density and side removal rate. The side removal rate increases with the increase of the duty ratio and lateral gap. Increasing of the duty ratio and decreasing of the lateral gap will increase the temperature and current density. Increasing the inlet pressure accelerates the frequency of renewal of heat and electrolysis products, which can reduce the single side gap. The experience of the ECM holes verifies the results of the simulation. The accuracy and stability of the ECM of holes are enhanced by optimizing the duty ratio, lateral gap and inlet pressure.

The effect of character stroke width on legibility: The relationship between duty ratio and contrast threshold.

Stroke width is an important factor affecting letter legibility. Although there seems to be a critical or optimal value for stroke width, it has been difficult to estimate quantitatively because shape deterioration and the increase of stroke width are covariates. This study attempted to predict the optimal value of stroke width considering it as a duty ratio. The duty ratio is related to the amplitude of the fundamental frequency, with the maximum amplitude of the fundamental frequency occurring at a duty ratio of 0.5. The fundamental frequency decreases when the duty ratio is set either lower or higher than 0.5. Based on previous research indicating that letters also have a critical band (CB), or the frequency band which contributes most to letter recognition, which corresponds to their fundamental frequency, we hypothesized that the amplitude of CB is related to the legibility of letters. We measured the contrast thresholds using square-wave gratings and letters of differing duty ratios. In both the grating and letter stimuli, the amplitude of CB reached its maximum when the duty ratio was around 0.5. Contrast thresholds for stimuli with various duty ratios were accurately predicted by the amplitudes of CB and the contrast thresholds in the individual's sine-wave grating. Our results suggest that the amplitude of a single critical component contributes significantly to the legibility of letters, even though they contain diverse frequency components.

Unraveling a Force-Generating Allosteric Pathway of Actomyosin Communication Associated with ADP and Pi Release.

The actomyosin system generates mechanical work with the execution of the power stroke, an ATP-driven, two-step rotational swing of the myosin-neck that occurs post ATP hydrolysis during the transition from weakly to strongly actin-bound myosin states concomitant with Pi release and prior to ADP dissociation. The activating role of actin on product release and force generation is well documented; however, the communication paths associated with weak-to-strong transitions are poorly characterized. With the aid of mutant analyses based on kinetic investigations and simulations, we identified the W-helix as an important hub coupling the structural changes of switch elements during ATP hydrolysis to temporally controlled interactions with actin that are passed to the central transducer and converter. Disturbing the W-helix/transducer pathway increased actin-activated ATP turnover and reduced motor performance as a consequence of prolonged duration of the strongly actin-attached states. Actin-triggered Pi release was accelerated, while ADP release considerably decelerated, both limiting maximum ATPase, thus transforming myosin-2 into a high-duty-ratio motor. This kinetic signature of the mutant allowed us to define the fractional occupancies of intermediate states during the ATPase cycle providing evidence that myosin populates a cleft-closure state of strong actin interaction during the weak-to-strong transition with bound hydrolysis products before accomplishing the power stroke.

Inactivation Kinetics and Membrane Potential of Pathogens in Soybean Curd Subjected to Pulsed Ohmic Heating Depending on Applied Voltage and Duty Ratio.

The aim of this research was to investigate the efficacy of the duty ratio and applied voltage in the inactivation of pathogens in soybean curd by pulsed ohmic heating (POH). The heating rate of soybean curd increased rapidly as the applied voltage increased, although the duty ratio did not affect the temperature profile. We supported this result by verifying that electrical conductivity increased with the applied voltage. Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes in soybean curd were significantly (P < 0.05) inactivated by more than 1 log unit at 80 Vrms (root mean square voltage). To elucidate the mechanism underlying these results, the membrane potential of the pathogens was examined using DiBAC4(3) [bis-(1,3-dibutylbarbituric acid)trimethine oxonol] on the basis of a previous study showing that the electric field generated by ohmic heating affected the membrane potential of cells. The values of DiBAC4(3) accumulation increased under increasing applied voltage, and they were significantly (P < 0.05) higher at 80 Vrms, while the duty ratio had no effect. In addition, morphological analysis via transmission electron microscopy showed that electroporation and expulsion of intracellular materials were predominant at 80 Vrms Moreover, electrode corrosion was overcome by the POH technique, and the textural and color properties of soybean curd were preserved. These results substantiate the idea that the applied voltage has a profound effect on the microbial inactivation of POH as a consequence of not only the thermal effect, but also the nonthermal effect, of the electric field, whereas the duty ratio does not have such an effect.IMPORTANCE High-water-activity food products, such as soybean curd, are vulnerable to microbial contamination, which causes fatal foodborne diseases and food spoilage. Inactivating microorganisms inside food is difficult because the transfer of thermal energy is slower inside than it is outside the food. POH is an adequate sterilization technique because of its rapid and uniform heating without causing electrode corrosion. To elucidate the electrical factors associated with POH performance in the inactivation of pathogens, the effects of the applied voltage and duty ratio on POH were investigated. In this study, we verified that a high applied voltage (80 Vrms) at a duty ratio of 0.1 caused thermal and nonthermal effects on pathogens that led to an approximately 4-log-unit reduction in a significantly short time. Therefore, the results of this research corroborate database predictions of the inactivation efficiency of POH based on pathogen control strategy modeling.

Direct observation of speed fluctuations of flagellar motor rotation at extremely low load close to zero.

The bacterial flagellar motor accommodates ten stator units around the rotor to produce large torque at high load. But when external load is low, some previous studies showed that a single stator unit can spin the rotor at the maximum speed, suggesting that the maximum speed does not depend on the number of active stator units, whereas others reported that the speed is also dependent on the stator number. To clarify these two controversial observations, much more precise measurements of motor rotation would be required at external load as close to zero as possible. Here, we constructed a Salmonella filament-less mutant that produces a rigid, straight, twice longer hook to efficiently label a 60 nm gold particle and analyzed flagellar motor dynamics at low load close to zero. The maximum motor speed was about 400 Hz. Large speed fluctuations and long pausing events were frequently observed, and they were suppressed by either over-expression of the MotAB stator complex or increase in the external load, suggesting that the number of active stator units in the motor largely fluctuates near zero load. We conclude that the lifetime of the active stator unit becomes much shorter when the motor operates near zero load.

Thick Filament Mechano-Sensing in Skeletal and Cardiac Muscles: A Common Mechanism Able to Adapt the Energetic Cost of the Contraction to the Task.

A dual regulation of contraction operates in both skeletal and cardiac muscles. The first mechanism, based on Ca2+-dependent structural changes of the regulatory proteins in the thin filament, makes the actin sites available for binding of the myosin motors. The second recruits the myosin heads from the OFF state, in which they are unable to split ATP and bind to actin, in relation to the force during contraction. Comparison of the relevant X-ray diffraction signals marking the state of the thick filament demonstrates that the force feedback that controls the regulatory state of the thick filament works in the same way in skeletal as in cardiac muscles: even if in an isometric tetanus of skeletal muscle force is under the control of the firing frequency of the motor unit, while in a heartbeat force is controlled by the afterload, the stress-sensor switching the motors ON plays the same role in adapting the energetic cost of the contraction to the force. A new aspect of the Frank-Starling law of the heart emerges: independent of the diastolic filling of the ventricle, the number of myosin motors switched ON during systole, and thus the energetic cost of contraction, are tuned to the arterial pressure. Deterioration of the thick-filament regulation mechanism may explain the hyper-contractility related to hypertrophic cardiomyopathy, an inherited heart disease that in 40% of cases is due to mutations in cardiac myosin.

Dilated cardiomyopathy myosin mutants have reduced force-generating capacity.

Dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM) can cause arrhythmias, heart failure, and cardiac death. Here, we functionally characterized the motor domains of five DCM-causing mutations in human ß-cardiac myosin. Kinetic analyses of the individual events in the ATPase cycle revealed that each mutation alters different steps in this cycle. For example, different mutations gave enhanced or reduced rate constants of ATP binding, ATP hydrolysis, or ADP release or exhibited altered ATP, ADP, or actin affinity. Local effects dominated, no common pattern accounted for the similar mutant phenotype, and there was no distinct set of changes that distinguished DCM mutations from previously analyzed HCM myosin mutations. That said, using our data to model the complete ATPase contraction cycle revealed additional critical insights. Four of the DCM mutations lowered the duty ratio (the ATPase cycle portion when myosin strongly binds actin) because of reduced occupancy of the force-holding A·M·D complex in the steady state. Under load, the A·M·D state is predicted to increase owing to a reduced rate constant for ADP release, and this effect was blunted for all five DCM mutations. We observed the opposite effects for two HCM mutations, namely R403Q and R453C. Moreover, the analysis predicted more economical use of ATP by the DCM mutants than by WT and the HCM mutants. Our findings indicate that DCM mutants have a deficit in force generation and force-holding capacity due to the reduced occupancy of the force-holding state.

Seeker-Azimuth Determination with Gyro Rotor and Optoelectronic Sensors.

This paper presents an approach to seeker-azimuth determination using the gyro rotor and optoelectronic sensors. In the proposed method, the gyro rotor is designed with a set of black and white right spherical triangle patterns on its surface. Two pairs of optoelectronic sensors are located symmetrically around the gyro rotor. When there is an azimuth, the stripe width covering the black and white patterns changes. The optoelectronic sensors then capture the reflected optical signals from the different black and white pattern stripes on the gyro rotor and produce the duty ratio signal. The functional relationship between the measured duty ratio and the azimuth information is numerically derived, and, based on this relationship, the azimuth is determined from the measured duty ratio. Experimental results show that the proposed approach produces a large azimuth range and high measurement accuracy with the linearity error of less than 0.005.

Cardiac actin changes in the actomyosin interface have different effects on myosin duty ratio.

Hypertrophic cardiomyopathy (HCM) is an inherited cardiovascular disease (CD) that commonly causes an increased size of cardiomyocytes in the left ventricle. The proteins myosin and actin interact in the myocardium to produce contraction through the actomyosin ATPase cycle. The duty ratio (r) of myosin is the proportion of the actomyosin ATPase cycle that myosin is bound to actin and does work. A common hypothesis is that HCM mutations increase contraction in cardiac sarcomeres; however, the available data are not clear on this connection. Based on previous work with human α-cardiac actin (ACTC), we hypothesize that HCM-linked ACTC variants with alterations near the myosin binding site have an increased r, producing more force. Myosin duty ratios using human ACTC variant proteins were calculated with myosin ATPase activity and in-vitro motility data. We found no consistent changes in the duty ratio of the ACTC variants, suggesting that other factors are involved in the development of HCM when ACTC variants are present.

A Rapid One-Step Process for Fabrication of Biomimetic Superhydrophobic Surfaces by Pulse Electrodeposition.

Inspired by some typical plants such as lotus leaves, superhydrophobic surfaces are commonly prepared by a combination of low surface energy materials and hierarchical micro/nano structures. In this work, superhydrophobic surfaces on copper substrates were prepared by a rapid, facile one-step pulse electrodepositing process, with different duty ratios in an electrolyte containing lanthanum chloride (LaCl3·6H2O), myristic acid (CH3(CH2)12COOH), and ethanol. The equivalent electrolytic time was only 10 min. The surface morphology, chemical composition and superhydrophobic property of the pulse electrodeposited surfaces were fully investigated with SEM, EDX, XRD, contact angle meter and time-lapse photographs of water droplets bouncing method. The results show that the as-prepared surfaces have micro/nano dual scale structures mainly consisting of La[CH3(CH2)12COO]3 crystals. The maximum water contact angle (WCA) is about 160.9°, and the corresponding sliding angle is about 5°. This method is time-saving and can be easily extended to other conductive materials, having a great potential for future applications.

Bed Rest and Hypoxic Exposure Affect Sleep Architecture and Breathing Stability.

Objective: Despite over 50 years of research on the physiological effects of sustained bed rest, data characterizing its effects on sleep macrostructure and breathing stability in humans are scarce. This study was conducted to determine the effects of continuous exposure to hypoxia and sustained best rest, both individually and combined, on nocturnal sleep and breathing stability. Methods: Eleven participants completed three randomized, counter-balanced, 21-days trials of: (1) normoxic bed rest (NBR, PIO2 = 133.1 ± 0.3), (2) hypoxic ambulatory confinement (HAMB, PIO2 = 90.0 ± 0.4) and (3) hypoxic bed rest (HBR, PIO2 = 90.0 ± 0.4; ~4,000 m equivalent altitude). Full objective polysomnography was performed at baseline, on Night 1 and Night 21 in each condition. Results: In NBR Night 1, more time was spent in light sleep (10 ± 2%) compared to baseline (8 ± 2%; p = 0.028); Slow-wave sleep (SWS) was reduced from baseline in the hypoxic-only trial by 18% (HAMB Night 21, p = 0.028) and further reduced by 33% (HBR Night 1, p = 0.010), and 36% (HBR Night 21, p = 0.008) when combined with bed rest. The apnea-hypopnea index doubled from Night 1 to Night 21 in HBR (32-62 events·h-1) and HAMB (31-59 events·h-1; p = 0.002). Those who experienced greatest breathing instability from Night 1 to Night 21 (NBR) were correlated to unchanged or higher (+1%) night SpO2 concentrations (R2 = 0.471, p = 0.020). Conclusion: Bed rest negatively affects sleep macrostructure, increases the apnea-hypopnea index, and worsens breathing stability, each independently exacerbated by continuous exposure to hypoxia.

Myosins: Domain Organisation, Motor Properties, Physiological Roles and Cellular Functions.

Myosins are cytoskeletal motor proteins that use energy derived from ATP hydrolysis to generate force and movement along actin filaments. Humans express 38 myosin genes belonging to 12 classes that participate in a diverse range of crucial activities, including muscle contraction, intracellular trafficking, cell division, motility, actin cytoskeletal organisation and cell signalling. Myosin malfunction has been implicated a variety of disorders including deafness, hypertrophic cardiomyopathy, Usher syndrome, Griscelli syndrome and cancer. In this chapter, we will first discuss the key structural and kinetic features that are conserved across the myosin family. Thereafter, we summarise for each member in turn its unique functional and structural adaptations, cellular roles and associated pathologies. Finally, we address the broad therapeutic potential for pharmacological interventions that target myosin family members.

Do cardiac actin mutations lead to altered actomyosin interactions?

It is currently hypothesized that increased heart muscle contractility leads to hypertrophic cardiomyopathy (HCM), and reduced contractility leads to dilated cardiomyopathy (DCM). To determine if changes in the core interaction between actin and myosin occur due to mutations in the cardiac actin gene (ACTC), we measured the interactions between myosin and 8 ACTC mutant proteins found in patients with HCM or DCM. R312H showed a decreased actin-activated myosin S1 ATPase rate (13.1 ± 0.63 µmol/L/min) compared to WT (15.3 ± 1.6 µmol/L/min), whereas the rate with E99K was significantly higher (20.1 ± 1.5 µmol/L/min). In vitro motility assays with varying ATP concentrations showed that the KM for E99K remains unchanged with a significantly decreased Vmax (1.90 ± 0.37 µm/sec) compared to WT (3.33 ± 0.46 µm/sec). Based on a 5 nm myosin step size, we calculated a duty ratio of approximately 0.04 for WT and the majority of mutant actins; however, the duty ratio for E99K was twice as high. Based on our analysis of 8 ACTC mutants, we infer that mutations in ACTC lead to disease through various molecular mechanisms. While changes in actomyosin interactions with the E99K mutation might cause increased ATP usage and tension leading to HCM, measurable changes in the basic interaction between actin and myosin do not appear to be involved in the mechanisms of disease development for the other ACTC mutants tested.

Getting myosin-V on the right track: tropomyosin sorts transport in yeast.

Recent studies have revealed a novel mechanism of myosin regulation in which the actin-binding protein tropomyosin converts atypical type-V myosins into processive cargo transporters. To achieve this, tropomyosin's primary role appears to lie in its ability to influence myosin's enzyme kinetics, prolonging the strong actin-bound ADP/apo state to enable hand-over-hand walking of myosin-V dimers along actin tracks. Activation of myosin-V mediated transport by tropomyosin underscores its function in helping to direct cargos to specific actin tracks and subcellular destinations. This type of regulation supports the broader notion that tropomyosin plays a key role in actomyosin sorting.