Cell-Engineered Recombinant α-Synuclein: A Gage R&R Validated Protocol.

α-Synuclein (α-Syn) misfolding and its presence in Lewy bodies are observed in almost all Parkinson's disease (PD) patients. Basic biomedical research would benefit from a quick, low-cost approach to purifying α-Syn and developing in vitro and in vivo models for PD. Several research groups utilize PFF-based models, yet the production of α-Syn PFFs is inconsistent, resulting in nonconclusive findings. Some research laboratories prepare recombinant α-Syn (r α-Syn) by molecular cloning to overexpress α-Syn with various purifying techniques. Laboratory-to-laboratory protocols cause considerable variability and sometimes contradictory findings. PD researchers spend more on protein than solving α-Syn's riddles. This article uncovered a novel method for expressing and purifying r α-Syn validated through gage reproducibility and repeatability (Gage R&R). For the production of r α-Syn, we have employed the ability of a high-cell-density-based expression system to overexpress protein in BL21(DE3). A simple, high-throughput, nonchromatographical purification protocol has been devised to facilitate research with higher reproducibility, which was validated through Gage R&R. A crossover experimental design was utilized, and the purified protein was characterized using orthogonal high-end analytical methods, which displayed higher similarity between the isolated r α-Syn. Batch-to-batch variability was the least for produced protein and hence can be utilized for exploring the iceberg of PD.

Smart Pneumatic Artificial Muscle Using a Bend Sensor like a Human Muscle with a Muscle Spindle.

Shortage of labor and increased work of young people are causing problems in terms of care and welfare of a growing proportion of elderly people. This is a looming social problem because people of advanced ages are increasing. Necessary in the fields of care and welfare, pneumatic artificial muscles in actuators of robots are being examined. Pneumatic artificial muscles have a high output per unit of weight, and they are soft, similarly to human muscles. However, in previous research of robots using pneumatic artificial muscles, rigid sensors were often installed at joints and other locations due to the robots' structures. Therefore, we developed a smart actuator that integrates a bending sensor that functions as a human muscle spindle; it can be externally attached to the pneumatic artificial muscle. This paper reports a smart artificial muscle actuator that can sense contraction, which can be applied to developed self-monitoring and robot posture control.

A Comprehensive Design of Six-Axis Force/Moment Sensor.

Strain gage type six-axis force/moment (F/M) sensors have been largely studied and implemented in industrial applications by using an external data acquisition board (DAQ). The use of external DAQs will ill-affect accuracy and crosstalk due to the possibility of voltage drop through the wire length. The most recent research incorporated DAQ within a relatively small F/M sensor, but only for sensors of the capacitance and optical types. This research establishes the integration of a high-efficiency DAQ on six-axis F/M sensor with a revolutionary arrangement of 32 strain gages. The updated structural design was optimized using the sequential quadratic programming method and validated using Finite Element Analysis (FEA). A new, integrated DAQ system was designed, tested, and compared to commercial DAQ systems. The proposed six-axis F/M sensor was examined with the calibrated jig. The results show that the measurement error and crosstalk have been significantly reduced to 1.15% and 0.68%, respectively, the best published combination at this moment.

Determining Wheel Forces and Moments on Aircraft Landing Gear with a Dynamometer Sensor.

This paper describes airfield measurement of forces and moments that act on a landing gear wheel. For the measurement, a wheel force sensor was used. The sensor was designed and built based on strain gage technology and was embedded in the left landing gear wheel of a test aircraft. The sensor is capable of measuring simultaneously three perpendicular forces and three moments and sends data to a handheld device wirelessly. For the airfield tests, the sensor was installed on a PZL 104 Wilga 35A multipurpose aircraft. The aircraft was towed at a "marching man" speed and the measurements were performed at three driving modes: Free rolling, braking, and turning. The paper contains results obtained in the field measurements performed on a grassy runway of the Rzeszów Jasionka Aerodrome, Poland. Rolling resistance of aircraft tire, braking friction, as well as aligning moment were analyzed and discussed with respect to surface conditions.

Upregulated circular RNA circ_0016760 indicates unfavorable prognosis in NSCLC and promotes cell progression through miR-1287/GAGE1 axis.

Non-small cell lung cancer (NSCLC) is an aggressive malignancy with increasing worldwide incidence and is characterized by dismal prognosis due to its early recurrence and metastasis. Accumulating evidence documented that aberrantly expressed circular RNAs (circRNAs) are critically involved in tumorigenesis. In the current study, we focused on a novel circRNA, circ_0016760 in NSCLC. qRT-PCR was carried out to determine the expression level of circ_0016760 in tissue samples and cell lines. The clinical value of circ_0016760 was also investigated. The functions of circ_0016760 was explored by CCK-8, colony formation, flow cytometry, Transwell and animal experiments. Luciferase reporter assays were conducted to investigate the association between circ_0016760 and miR-1287 and miR-1287 and GAGE1. We found that circ_0016760 was enhanced in NSCLC tissues and cells and the upregulation of circ_0016760 is associated with advanced TNM stages, lymph node metastasis and adverse prognosis in NSCLC patients. Moreover, circ_0016760 could significantly promote cell growth and metastatic properties and inhibit cell apoptosis in NSCLC cells. In mechanism, circ_0016760 functioned as a sponge for miR-1287 and regulated the expression of GAGE1. Subsequently, functional assays illustrated that the oncogenic properties of circ_0016760 is partly attribute to the regulation of miR-1287/GAGE1 axis. In summary, circ_0016760/miR-1287/GAGE1 signaling might play vital roles in the tumorigenesis and progression of NSCLC.

Distributed Deformation Monitoring for a Single-Cell Box Girder Based on Distributed Long-Gage Fiber Bragg Grating Sensors.

Distributed deformation based on fiber Bragg grating sensors or other kinds of strain sensors can be used to monitor bridges during operation. However, most research on distributed deformation monitoring has focused on solid rectangular beams rather than box girders-a kind of typical hollow beam widely employed in actual bridges. The deformation of a single-cell box girder contains bending deflection and also two additional deformations respectively caused by shear lag and shearing action. This paper revises the improved conjugated beam method (ICBM) based on the long-gage fiber Bragg grating (LFBG) sensors to satisfy the requirements for monitoring the two additional deformations in a single-cell box girder. This paper also proposes a suitable LFBG sensor placement in a box girder to overcome the influence of strain fluctuation on the flange caused by the shear lag effect. Results from numerical simulations show that the theoretical monitoring errors of the revised ICBM are typically 0.3⁻1.5%, and the maximum error is 2.4%. A loading experiment for a single-cell box gilder monitored by LFBG sensors shows that most of the practical monitoring errors are 6⁻8% and the maximum error is 11%.

Bragg-Grating-Based Photonic Strain and Temperature Sensor Foils Realized Using Imprinting and Operating at Very Near Infrared Wavelengths.

Thin and flexible sensor foils are very suitable for unobtrusive integration with mechanical structures and allow monitoring for example strain and temperature while minimally interfering with the operation of those structures. Electrical strain gages have long been used for this purpose, but optical strain sensors based on Bragg gratings are gaining importance because of their improved accuracy, insusceptibility to electromagnetic interference, and multiplexing capability, thereby drastically reducing the amount of interconnection cables required. This paper reports on thin polymer sensor foils that can be used as photonic strain gage or temperature sensors, using several Bragg grating sensors multiplexed in a single polymer waveguide. Compared to commercially available optical fibers with Bragg grating sensors, our planar approach allows fabricating multiple, closely spaced sensors in well-defined directions in the same plane realizing photonic strain gage rosettes. While most of the reported Bragg grating sensors operate around a wavelength of 1550 nm, the sensors in the current paper operate around a wavelength of 850 nm, where the material losses are the lowest. This was accomplished by imprinting gratings with pitches 280 nm, 285 nm, and 290 nm at the core-cladding interface of an imprinted single mode waveguide with cross-sectional dimensions 3 × 3 µm². We show that it is possible to realize high-quality imprinted single mode waveguides, with gratings, having only a very thin residual layer which is important to limit bend losses or cross-talk with neighboring waveguides. The strain and temperature sensitivity of the Bragg grating sensors was found to be 0.85 pm/µÎµ and -150 pm/°C, respectively. These values correspond well with those of previously reported sensors based on the same materials but operating around 1550 nm, taking into account that sensitivity scales with the wavelength.

Transient Analysis of a Circular Foil Gage in a Convective and Radiative Environment.

An analysis of a circular thin-foil gage is presented that includes transient effects, convective heat transfer, and an arbitrary time-varying boundary condition at the foil edge to account for fluctuations in cooling water temperature. The governing energy equation is solved using Laplace transform to obtain the temporal and spatial temperature distributions of the foil. Under constant temperature at the foil edge and constant thermal radiative heat flux, closed-form response curves for the gage under various modes of heat transfer are provided. Steady-state results are also presented as limiting cases.

The unique C- and N-terminal sequences of Metallothionein isoform 3 mediate growth inhibition and Vectorial active transport in MCF-7 cells.

BACKGROUND: The 3rd isoform of the metallothionein (MT3) gene family has been shown to be overexpressed in most ductal breast cancers. A previous study has shown that the stable transfection of MCF-7 cells with the MT3 gene inhibits cell growth. The goal of the present study was to determine the role of the unique C-terminal and N-terminal sequences of MT3 on phenotypic properties and gene expression profiles of MCF-7 cells. METHODS: MCF-7 cells were transfected with various metallothionein gene constructs which contain the insertion or the removal of the unique MT3 C- and N-terminal domains. Global gene expression analysis was performed on the MCF-7 cells containing the various constructs and the expression of the unique C- and N- terminal domains of MT3 was correlated to phenotypic properties of the cells. RESULTS: The results of the present study demonstrate that the C-terminal sequence of MT3, in the absence of the N-terminal sequence, induces dome formation in MCF-7 cells, which in cell cultures is the phenotypic manifestation of a cell's ability to perform vectorial active transport. Global gene expression analysis demonstrated that the increased expression of the GAGE gene family correlated with dome formation. Expression of the C-terminal domain induced GAGE gene expression, whereas the N-terminal domain inhibited GAGE gene expression and that the effect of the N-terminal domain inhibition was dominant over the C-terminal domain of MT3. Transfection with the metallothionein 1E gene increased the expression of GAGE genes. In addition, both the C- and the N-terminal sequences of the MT3 gene had growth inhibitory properties, which correlated to an increased expression of the interferon alpha-inducible protein 6. CONCLUSIONS: Our study shows that the C-terminal domain of MT3 confers dome formation in MCF-7 cells and the presence of this domain induces expression of the GAGE family of genes. The differential effects of MT3 and metallothionein 1E on the expression of GAGE genes suggests unique roles of these genes in the development and progression of breast cancer. The finding that interferon alpha-inducible protein 6 expression is associated with the ability of MT3 to inhibit growth needs further investigation.

Suitability of Strain Gage Sensors for Integration into Smart Sport Equipment: A Golf Club Example.

Wearable devices and smart sport equipment are being increasingly used in amateur and professional sports. Smart sport equipment employs various sensors for detecting its state and actions. The correct choice of the most appropriate sensor(s) is of paramount importance for efficient and successful operation of sport equipment. When integrated into the sport equipment, ideal sensors are unobstructive, and do not change the functionality of the equipment. The article focuses on experiments for identification and selection of sensors that are suitable for the integration into a golf club with the final goal of their use in real time biofeedback applications. We tested two orthogonally affixed strain gage (SG) sensors, a 3-axis accelerometer, and a 3-axis gyroscope. The strain gage sensors are calibrated and validated in the laboratory environment by a highly accurate Qualisys Track Manager (QTM) optical tracking system. Field test results show that different types of golf swing and improper movement in early phases of golf swing can be detected with strain gage sensors attached to the shaft of the golf club. Thus they are suitable for biofeedback applications to help golfers to learn repetitive golf swings. It is suggested that the use of strain gage sensors can improve the golf swing technical error detection accuracy and that strain gage sensors alone are enough for basic golf swing analysis. Our final goal is to be able to acquire and analyze as many parameters of a smart golf club in real time during the entire duration of the swing. This would give us the ability to design mobile and cloud biofeedback applications with terminal or concurrent feedback that will enable us to speed-up motor skill learning in golf.

GAGE12 mediates human gastric carcinoma growth and metastasis.

The spontaneous metastasis from human gastric carcinoma (GC) remains poorly reproduced in animal models. Here, we established an experimental mouse model in which GC progressively developed in the orthotopic stomach wall and metastasized to multiple organs; the tumors colonized in the ovary exhibited typical characteristics of Krukenberg tumor. The expression of mesenchymal markers was low in primary tumors and high in those in intravasating and extravasating veins. However, the expression of epithelial markers did not differ, indicating that the acquisition of mesenchymal markers without a concordant loss of typical epithelial markers was associated with metastasis. We identified 35 differentially expressed genes (DEGs) in GC cells metastasized to ovary, among which overexpression of GAGE12 family genes, the top-ranked DEGs, were validated. In addition, knockdown of the GAGE12 gene family affected transcription of many of the aforementioned 35 DEGs and inhibited trans-well migration, tumor sphere formation in vitro and tumor growth in vivo. In accordance, GAGE12 overexpression augmented migration, tumor sphere formation and sustained in vivo tumor growth. Taken together, the GAGE12 gene family promotes GC growth and metastasis by modulating the expression of GC metastasis-related genes.

Review of Trackside Monitoring Solutions: From Strain Gages to Optical Fibre Sensors.

A review of recent research on structural monitoring in railway industry is proposed in this paper, with a special focus on stress-based solutions. After a brief analysis of the mechanical behaviour of ballasted railway tracks, an overview of the most common monitoring techniques is presented. A special attention is paid on strain gages and accelerometers for which the accurate mounting position on the track is requisite. These types of solution are then compared to another modern approach based on the use of optical fibres. Besides, an in-depth discussion is made on the evolution of numerical models that investigate the interaction between railway vehicles and tracks. These models are used to validate experimental devices and to predict the best location(s) of the sensors. It is hoped that this review article will stimulate further research activities in this continuously expanding field.

Exploring quantitative indices to characterize piano timbre with precision validated using measurement system analysis.

Aim: Timbre in piano performance plays a critical role in enhancing musical expression. However, timbre control in current piano performance education relies mostly on descriptive characterization, which involves large variations of interpretation. The current study aimed to mitigate the limitations by identifying quantitative indices with adequate precision to characterize piano timbre. Methods: A total of 24 sounds of G6 were recorded from 3 grand pianos, by 2 performers, and with 4 repetitions. The sounds were processed and analyzed with audio software for the frequencies and volumes of harmonic series in the spectrum curves. Ten quantitative timbre indices were calculated. Precision validation with statistical gage R&R analysis was conducted to gage the repeatability (between repetitions) and reproducibility (between performers) of the indices. The resultant percentage study variation (%SV) of an index must be ≤10% to be considered acceptable for characterizing piano timbre with enough precision. Results: Out of the 10 indices, 4 indices had acceptable precision in characterizing piano timbre with %SV ≤10%, including the square sum of relative volume (4.40%), the frequency-weighted arithmetic mean of relative volume (4.29%), the sum of relative volume (3.11%), and the frequency-weighted sum of relative volume (2.09%). The novel indices identified in the current research will provide valuable tools to advance the measurement and communication of timbre and advance music performance education.

Evaluation of accuracy of direct transfer snapon impression coping closed tray impression technique and direct transfer open tray impression technique: an in vitro study.

Accuracy of the implant impression technique is one of the key factor determining the strain free fit of the prosthesis fabricated which influences the treatment success. Two implant impression techniques namely the closed tray technique with transfer coping and open tray technique were evaluated for accuracy with stone casts obtained from them. Casts were evaluated using a custom constructed bar on strain gage (SYSCOM) and abutment coordinates using Coordinate Measuring Machine (TESA micro-HITE). The statistical analysis with one way ANOVA and Mann-Whitney tests show that the casts obtained with open tray technique were accurate than the casts of closed tray technique (significance P < 0.001). Direct transfer impression technique with less number of components ensures the high accuracy of transfer of implant positions from master cast to the laboratory cast compared to the indirect transfer impression technique.

Sagittal suture strain in capuchin monkeys (Sapajus and Cebus) during feeding.

OBJECTIVES: Morphological variation in cranial sutures is used to infer aspects of primate feeding behavior, including diet, but strain regimes across sutures are not well documented. Our aim is to test hypotheses about sagittal suture morphology, strain regime, feeding behavior, and muscle activity relationships in robust Sapajus and gracile Cebus capuchin primates. MATERIALS AND METHODS: Morphometrics of sinuosity in three regions of the sagittal suture were compared among museum specimens of Sapajus and Cebus, as well as in robust and gracile lab specimens. In vivo strains and bilateral electromyographic (EMG) activity were recorded from these regions in the temporalis muscles of capuchin primates while they fed on mechanically-varying foods. RESULTS: Sapajus and the anterior suture region exhibited greater sinuosity than Cebus and posterior regions. In vivo data reveal minor differences in strain regime between robust and gracile phenotypes but show higher strain magnitudes in the middle suture region and higher tensile strains anteriorly. After gage location, feeding behavior has the most consistent and strongest impact on strain regime in the sagittal suture. Strain in the anterior suture has a high tension to compression ratio compared to the posterior region, especially during forceful biting in the robust Sapajus-like individual. DISCUSSION: Sagittal suture complexity in robust capuchins likely reflects feeding behaviors associated with mechanically challenging foods. Sutural strain regimes in other anthropoid primates may also be affected by activity in feeding muscles.

Investigation of the transient behavior of a cantilever beam using PVDF sensors.

In this paper, a PVDF film sensor was used to measure the transient responses of a cantilever beam subjected to an impact loading. The measurement capability of a PVDF sensor is affected by the area of the PVDF film sensor and the signal conditioner (charge amplifier). The influences of these effects on the experimental measurements were investigated. The transient responses for the dynamic strain of the beam were measured simultaneously by the PVDF sensor and a conventional strain gauge. The resonant frequencies of the beam were determined by applying the Fast Fourier Transform on transient results in the time domain of the PVDF sensor and the strain gauge. The experimentally measured resonant frequencies from the PVDF sensor and the strain gauge were compared with those predicted from theoretical and FEM numerical calculations. Based on the comparison of the results measured for these two sensors, the PVDF film sensor proved capable of measuring transient responses for dynamic strain, and its sensitivity is better than that of the strain gauge. Furthermore, almost all the resonant frequencies can be obtained from the results of transient responses for PVDF film.

Computational Micromechanics Investigation of Percolation and Effective Electro-Mechanical Properties of Carbon Nanotube/Polymer Nanocomposites using Stochastically Generated Realizations: Effects of Orientation and Waviness.

The electrical and mechanical properties of carbon nanotube/polymer nanocomposites depend strongly upon several factors such as CNT volume fraction, CNT alignment, CNT dispersion and CNT waviness among others. This work focuses on obtaining estimates and distribution for the effective electrical conductivity, elastic constants and piezoresistive properties as a function of these factors using a stochastic approach with numerous CNT/polymer realizations coupled with parallel computation. Additionally, electrical percolation volume fraction and percolation transitional behavior is also studied. The effective estimates and percolation values were found to be in good agreement with experimental works in the literature. It was found that with increasing CNT volume fraction, the mechanical properties improved. However, due to the interaction of CNTs with one another through electrical tunneling, the conductivity and piezoresistivity properties evolved in a more complex manner. While the degree of alignment played a strong role in the effective properties making them anisotropic, the effect of waviness was found to be insubstantial.

In the shadow of the dam - Hydrology of the Little Conemaugh river and its South Fork, with insights about past and future flooding.

The Little Conemaugh River watershed and its South Fork sub-basin figure prominently in historical flooding of Johnstown, Pennsylvania and nearby communities with catastrophic flooding in 1889, 1936, and 1977 (reviewed herein). Historical stream gage data and data from a new gage on the South Fork (established via a novel, portable cableway system) are used with Nexrad rainfall data to assess watershed response and provide novel analysis of flood hydrology in the Little Conemaugh basin and the sub-basin. Using unit hydrograph estimates for longer duration storms (>8 h) and different baseflow conditions, we probe possible effects of several design storms, including those stemming from a hurricane remnant scenario (Agnes in 1972) and 50-, 100-, and 500-year 12-hour precipitation depths. The unit hydrographs provided peak discharge (Qpeak) estimates for 1977 (the only flood event with available hourly rainfall data) that are in good agreement with empirical peak discharges. Significant channel improvements completed in 1943 were designed to carry the largest known natural flow on record at that time (1936 Qpeak). Preliminary results from design storm scenarios indicate the need for a careful evaluation of extreme discharges and their return periods (including snowmelt-related contributions), as future flood levels in Johnstown may occur more frequently than originally thought. The 1977 flood, which triggered 7 dam failures and eclipsed 1936 Qpeak, resulted from less than 40% the estimated probable maximum precipitation (PMP) for a 12-hour storm. Peak discharges of similar magnitude would have ensued in 1972 had remnants of Hurricane Agnes tracked slightly westward. Flooding and infrastructure problems could be compounded for storms of 24-hour or longer durations, similar to record flooding seen in central Pennsylvania and New York in 1972. Flood recurrence, emergency procedures, and dam safety (particularly, spillway capacity in the Little Conemaugh basin and surrounding region) should likely be reassessed and protective early-warning measures (ineffective in 1977) implemented for the people of Johnstown.

A method for assessing and monitoring consistency of nonclinical ECG analysis.

Cardiovascular safety is a key area of concern for new drugs in development, and the collection and analysis of electrocardiograms (ECGs) is a standard and major component of nonclinical testing. Digital data capture technology allows for high-throughput and long-duration ECG collections, resulting in large volumes of data. Consistent analysis of these ECG data is critical for detecting meaningful changes during nonclinical drug development. We developed a method to assess the consistency of nonclinical ECG analysis for a group of analysts over time. Eight­lead ECGs were collected from conscious dogs using Ponemah (v5.2, DSI). Analysts placed Pstart, Qstart, Rpeak, Send, and Tend marks on six waveforms for each animal. The ECG files were randomized and re-marked under blinded conditions 4 to 14 days following initial mark placement. Averages of each parameter measured (RR interval, QRS duration, PR interval, and QT interval) were compiled for each marking session and analyst. A Gage R&R evaluation was completed. Graphical output from the Gage R&R evaluation showed distinct variability on group and individual analyst levels. Differences in inter- and intra-analyst variability (reproducibility and repeatability, respectively) were observed between trained analysts and analysts in training. The Gage R&R method is an effective tool for assessing consistency of digital ECG mark placement at a group level. Furthermore, it is able to identify areas of improvement for individual ECG analysts and to assess ECG analyst consistency during their training period. The assessment results are useful for facilitating discussions on best practices and maintaining consistency of mark placement.

Supporting data for strengthening and deformation behavior of as-cast CoCrCu1.5MnNi high entropy alloy with micro-/nanoscale precipitation.

The data presented here are related to the research article entitled "Strengthening and deformation behavior of as-cast CoCrCu1.5MnNi high-entropy alloy (HEA) with micro-/nanoscale precipitation [1]". Non-equimolar CoCrCu1.5MnNi was cast by the conventional induction melting under a high-purity Ar atmosphere. Scanning electron microscopy equipped with energy dispersive spectroscopy (EDS), and transmission electron microscopy (TEM) were used for micro- and nanostructure characterization. Subsize tensile specimens with two different gage length to width ratio were tested at room and cryogenic temperatures to assess the accuracy of strength and ductility data in the as-cast CoCrCu1.5MnNi HEAs. The mixing enthalpy (ΔHmix) versus lattice elastic energy (ΔHel) criterion was used to predict the stable phases. The data on the effects of microstructural and nanostructural distribution of various phases on mechani-cal properties in the as-cast HEA could be used in designing high entropy alloys with excellent as-cast mechanical performance.