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Background The construction and application of synthetic genetic circuits is frequently improved if gene expression can be orthogonally controlled, relative to the host. In plants, orthogonality can be achieved via the use of CRISPR-based transcription factors that are programmed to act on natural or synthetic promoters. The construction of complex gene circuits can require multiple, orthogonal regulatory interactions, and this in turn requires that the full programmability of CRISPR elements be adapted to non-natural and non-standard promoters that have few constraints on their design. Therefore, we have developed synthetic promoter elements in which regions upstream of the minimal 35S CaMV promoter are designed from scratch to interact via programmed gRNAs with dCas9 fusions that allow activation of gene expression. Results A panel of three, mutually orthogonal promoters that can be acted on by artificial gRNAs bound by CRISPR regulators were designed. Guide RNA expression targeting these promoters was in turn controlled by either Pol III (U6) or ethylene-inducible Pol II promoters, implementing for the first time a fully artificial Orthogonal Control System (OCS). Following demonstration of the complete orthogonality of the designs, the OCS was tied to cellular metabolism by putting gRNA expression under the control of an endogenous plant signaling molecule, ethylene. The ability to form complex circuitry was demonstrated via the ethylene-driven, ratiometric expression of fluorescent proteins in single plants. Conclusions The design of synthetic promoters is highly generalizable to large tracts of sequence space, allowing Orthogonal Control Systems of increasing complexity to potentially be generated at will. The ability to tie in several different basal features of plant molecular biology (Pol II and Pol III promoters, ethylene regulation) to the OCS demonstrates multiple opportunities for engineering at the system level. Moreover, given the fungibility of the core 35S CaMV promoter elements, the derived synthetic promoters can potentially be utilized across a variety of plant species.

Electrodiagnostic studies (EDXs) are the confirmative diagnostic tool for carpal tunnel syndrome (CTS). Previous studies have evaluated the relationship between EDXs and ultrasonography (US) but not with X-rays. Recently, many studies on the diagnostic value of X-rays in various diseases have been reported, but data on CTS are lacking. We evaluated the relationship between electrodiagnostic parameters and roentgenographic and ultrasonographic features in CTS and investigated the usefulness of X-rays and US for CTS. This retrospective study included 97 wrists of 62 patients. All patients with suspected CTS underwent EDXs, wrist US, and wrist X-rays. The CTS patients were classified into mild, moderate, and severe groups. The roentgenographic features included the ulnar variance (UV) and the anteroposterior diameter of the wrist (APDW), and the ultrasonographic features included the flattening ratio (FR) and the thickest anteroposterior diameter of the median nerve (TAPDM). Most EDX parameters showed significant correlations with roentgenographic and US features. The electrodiagnostic severity was also correlated with all imaging features. Therefore, both wrist X-rays and wrist US can be useful for the diagnosis of CTS as supplements to EDXs.

Ticks transmit a diverse array of microbes to vertebrate hosts, including human pathogens, which has led to a human-centric focus in this vector system. Far less is known about pathogens of ticks themselves. Here, we discover that a toxin in blacklegged ticks (Ixodes scapularis) horizontally acquired from bacteria-called domesticated amidase effector 2 (dae2)-has evolved to kill mammalian skin microbes with remarkable efficiency. Secreted into the saliva and gut of ticks, Dae2 limits skin-associated staphylococci in ticks while feeding. In contrast, Dae2 has no intrinsic ability to kill Borrelia burgdorferi, the tick-borne Lyme disease bacterial pathogen. These findings suggest ticks resist their own pathogens while tolerating symbionts. Thus, just as tick symbionts can be pathogenic to humans, mammalian commensals can be harmful to ticks. Our study underscores how virulence is context-dependent and bolsters the idea that "pathogen" is a status and not an identity.

Lifelogging records personal data generated from one’s own physical and emotional activities in daily life. It is widely used to discover sleep, exercise, or socializing patterns. In recent years, wearable devices and mobile applications have been used to automatically log daily activities, expanding the capabilities of lifelogging applications for a variety of purposes. One such application is the development of self-awareness, comprehensive knowledge about the self with which one may discover their inclinations and set clear goals to enhance future performance. This study proposes an approach for improving self-awareness that allows users to precisely review personal standards and levels of happiness and stress using a smartwatch and mobile application. Related work is investigated to understand the various types of methods and purposes involved, and a survey is conducted to gain insights on daily activities that induce specific emotional states. The findings support dividing one’s daily activities into seven categories, which are socialization, leisure, eating, sleeping, working, studying, and other, and an effective mobile application is thus designed. The potential for behavioral changes and the positive effects of the gamification of data visualization to be deployed in the mobile application are also discussed.

This article deals with an ADAS (Advanced Driver Assistance System) application using newly developed neuromorphic artificial intelligent chip NM500. Neuromorphic artificial intelligence is distinguished from other AI by its particular hardware structure and parallel algorithms of learning and recognition. Thus, neurons of NM500 can learn and recognize patterns extracted from any data sources with less energy and complexity than modern microprocessors. The proposed application can control the vehicle speed by recognizing the traffic information images marked on road. We have built a small-scaled vehicle model to discuss the real-time performance as well as hardware implementation with NM500. Taking advantages of NM500, the system simply consists of a low-priced surveillance camera attached in the front windshield of a vehicle and an Arduino kit, which processes the video signal from the camera and speed control signal.

Current wooden reel modules are frequently used to carry high-voltage electric transmission lines by winding them around the reel module. But the wooden reel has been reported to have several problems for users and manufacturers, such as high manufacturing cost, heavy structure, and difficulty in recycling, so that it is necessary to make them from other light-weight materials with sufficient strength. One of the alternative materials is engineering plastic, and a new design process for reel modules should be developed for the engineering plastic material to minimize the total development duration. In this study, a numerical approach using a finite-element model of the reel module was used to ensure structural rigidity requirement over the maximum payload, 4,000 kgf, as well as the equivalent impact load from more than 10-cm free fall. The candidate finite-element model of the reel module was simulated for more than the maximum payload, and a structural static analysis was conducted for the equivalent impact force derived from free-fall motion with the center of the reel module fixed. Both simulation results revealed that the candidate reel model made from engineering plastic has satisfactory static rigidity compared with the current wooden reel module.

The intracellular bacterial pathogen Francisella tularensis causes tularemia, a zoonosis that can be fatal. The type VI secretion system (T6SS) encoded by the Francisella pathogenicity island (FPI) is critical for the virulence of this organism. Existing studies suggest that the complete repertoire of T6SS effectors delivered to host cells is encoded by the FPI. Using a proteome-wide approach, we discovered that the FPI-encoded T6SS exports at least three effectors encoded outside of the island. These proteins share features with virulence determinants of other pathogens, and we provide evidence that they can contribute to intramacrophage growth. The remaining proteins that we identified are encoded within the FPI. Two of these FPI-encoded proteins constitute effectors, whereas the others form a unique complex required for core function of the T6SS apparatus. The discovery of secreted effectors mediating interactions between Francisella and its host significantly advances our understanding of the pathogenesis of this organism.

This paper discusses about analyzing in-wheel vehicle’s dynamic motion and load torque. Since in-wheel vehicle controls each left and right driving wheels, it is dangerous if vehicle’s wheels are not in a cooperative control. First, this study builds the main wheel control logic using PID control theory and evaluates the stability. Using Carsim-Matlab/Simulink, vehicle dynamic motion is simulated in virtual 3D driving road. Through this, in-wheel vehicle’s driving performance can be analyzed. The target vehicle is a rear-wheel drive in D-class sedan. Second, by using the first In-wheel vehicle’s performance results, it derivate the drive motor’s dynamic load torque for applying the dynamometer. Extracted load torque impute to dynamometer’s load motor, linear experiment in dynamometer can replicated the 3-D road driving status. Also it, will be able to evaluate the more accurate performance analysis and stability, as a previous step of actual vehicle experiment. Key words : Inwheel(인휠), Carsim(카심), Driving performance(주행 성능), Load torque(부하 토크), Dynamo-meter(동력 시험계)

Although celecoxib is quite effective in the management of inflammation-related diseases, especially arthritis, its use is limited by concerns including low bioavailability (BA), non-linear pharmacokinetic (PK) profile, and peak concentration-related toxicity. To overcome these issues, we designed and prepared hydrophilic celecoxib prodrugs, namely N-glycyl-aspart-1yl celecoxib (N-GA1C), glutam-1-yl celecoxib (G1C), and aspart-1yl celecoxib (A1C), for the sustained release of celecoxib in the intestine with limited systemic absorption. The celecoxib derivatives were converted to celecoxib in the intestinal contents. The conversion rates were in order of N-GA1C &gt, G1C &gt, A1C. Oral administration of the celecoxib derivatives (oral celecoxib derivatives) sustained the plasma concentration of celecoxib for 24 h, improving the BA and linearity of the PK profile of celecoxib. The peak concentrations (Cmax) of celecoxib after oral celecoxib derivatives were lower than that after oral celecoxib. In a carrageenan-induced rat paw edema model, oral N-GA1C exhibited greater anti-inflammatory activity for a longer duration compared with oral celecoxib. The order of efficacy of the celecoxib derivatives was N-GA1C &gt, A1C. Taken together, the prodrug approach is a feasible strategy to improve the PK and therapeutic properties of celecoxib, and among the celecoxib derivatives, N-GA1C may be the most promising prodrug of celecoxib.

Non-holonomy in input-output systems can be characterized by a reachable space whose dimension is larger than that of its input space. Among the various mechanical examples of non-holonomy, we have dealt with the friction drive type semi-spherical continuously variable transmission (SS-CVT) in this paper. SS-CVT transmits power by rolling friction and realizes the neutral, forward and reverse states of output shaft without any additional devices. A spherical shaped variator in SS-CVT transmits power and changes its gear ratio keeping contact with an output disc. It is necessary to consider the friction forces along with in two-directional slip on the contact point, because the directions of friction forces to transmit power and to change gear ratio are perpendicular to each other. For this, the friction forces must be analyzed by using the slip ratio and slip angle on the contact patch. Therefore, we have determined the planar friction variables by the dimensional analysis, and the SS-CVT test bench. Finally, the empirical formula for evaluating the two friction forces has been proposed based on the experimental results. And the numerical results regarding the dynamics of SS-CVT have been addressed.

Bacteria in polymicrobial habitats contend with a persistent barrage of competitors, often under rapidly changing environmental conditions 1 . The direct antagonism of competitor cells is thus an important bacterial survival strategy 2 . Towards this end, many bacterial species employ an arsenal of antimicrobial effectors with multiple activities; however, the benefits conferred by the simultaneous deployment of diverse toxins are unknown. Here we show that the multiple effectors delivered to competitor bacteria by the type VI secretion system (T6SS) of Pseudomonas aeruginosa display conditional efficacy and act synergistically. One of these effectors, Tse4, is most active in high-salinity environments and synergizes with effectors that degrade the cell wall or inactivate intracellular electron carriers. We find Tse4 synergizes with these disparate mechanisms by forming pores that disrupt the ΔΨ component of the proton motive force. Our results provide evidence that the concomitant delivery of a cocktail of effectors serves as a bet-hedging strategy to promote bacterial competitiveness in the face of unpredictable and variable environmental conditions.

The Firmicutes are a phylum of bacteria that dominate numerous polymicrobial habitats of importance to human health and industry. Although these communities are often densely colonized, a broadly distributed contact-dependent mechanism of interbacterial antagonism utilized by Firmicutes has not been elucidated. Here we show that proteins belonging to the LXG polymorphic toxin family present in Streptococcus intermedius mediate cell contact- and Esx secretion pathway-dependent growth inhibition of diverse Firmicute species. The structure of one such toxin revealed a previously unobserved protein fold that we demonstrate directs the degradation of a uniquely bacterial molecule required for cell wall biosynthesis, lipid II. Consistent with our functional data linking LXG toxins to interbacterial interactions in S. intermedius, we show that LXG genes are prevalent in the human gut microbiome, a polymicrobial community dominated by Firmicutes. We speculate that interbacterial antagonism mediated by LXG toxins plays a critical role in shaping Firmicute-rich bacterial communities. DOI: http://dx.doi.org/10.7554/eLife.26938.001, eLife digest Most bacteria live in densely colonized environments, such as the human gut, in which they must constantly compete with other microbes for space and nutrients. As a result, bacteria have evolved a wide array of strategies to directly fight their neighbors. For example, some bacteria release antimicrobial compounds into their surroundings, while others ‘inject’ protein toxins directly into adjacent cells. Bacteria can be classified into two groups known as Gram-positive and Gram-negative. Previous studies found that Gram-negative bacteria inject toxins into neighboring cells, but no comparable toxins in Gram-positive bacteria had been identified. Before a bacterium can inject molecules into an adjacent cell, it needs to move the toxins from its interior to the cell surface. It had been suggested that a transport system in Gram-positive bacteria called the Esx pathway may export toxins known as LXG proteins. However, it was not clear whether these proteins help Gram-positive bacteria to compete against other bacteria. Whitney et al. studied the LXG proteins in Gram-positive bacteria known as Firmicutes. The experiments reveal that Firmicutes found in the human gut possess LXG genes. A Firmicute known as Streptococcus intermedius produces three LXG proteins that are all toxic to bacteria. To avoid being harmed by its own LXG proteins, S. intermedius also produces matching antidote proteins. Further experiments show that LXG proteins are exported out of S. intermedius cells and into adjacent competitor bacteria by the Esx pathway. Examining one of these LXG proteins in more detail showed that it can degrade a molecule that bacteria need to make their cell wall. Together, these findings suggest that LXG proteins may influence the species living in many important microbial communities, including the human gut. Changes in the communities of gut microbes have been linked with many diseases. Therefore, understanding more about how the LXG proteins work may help us to develop ways to manipulate these communities to improve human health. DOI: http://dx.doi.org/10.7554/eLife.26938.002

This article describes the dynamic behaviors of a rubber tired gantry crane(RTGC) under typical load conditions which are used in the design of gantry cranes. In order to investigate the dynamic characteristics of an RTGC, we developed a finite element crane model for its huge structure. The finite element model was validated with the modal test results, e.g., natural frequencies and normal modes. And other components of RTGC were converted into detailed 3D CAD models and finally transformed to rigid body models in a dynamic simulation program ADAMS. The load conditions considered in this paper were a normal operating condition(OP1) and container hanging condition with no external loads. As a result, we could find there was large influence of crane's vibration owing to its structural stiffness and deformation. And the vibration of crane could made the movements of RTGC, which occurs crash or malfunction of crane works. Keywords :rubber tired gantry crane, finite element, multi-flexible body dynamics, simulation, wheel load

This paper deals with the virtual prototyping and performance analysis of an electric vehicle powertrain using testbench and ADAMS software. The powertrain in this paper is proposed for use in an electric vehicle and especially adopts the function of an infinitely variable transmission (IVT). The IVT can realize not only a continuously variable transmission (CVT) function but also the forward, reverse, and neutral states of the vehicle speed without a clutch and/or brake. In this paper, a novel powertrain configuration is designed using an epicyclic gear set which can realize the IVT function by controlling the angular velocity of its one element. The virtual prototyping using ADAMS software is described in order to analyze the power performance of the proposed electric vehicle. And the dynamic behaviors of the electric vehicle are simulated and investigated with various driving conditions focused on the power performances. The simulation results are verified with the experiments of the testbench.

A method for the prediction of strain responses in a linear beam system using only output data—without direct measurement of stress or strain—is suggested. An indirect method is evaluated for predicting the strain behavior at a “hot spot” on a notched simple beam using the frequency response function (FRF) relating strain and acceleration. The dynamic response of the simple beam is obtained through uniaxial excitation. The results show that the prediction of dynamic strain is accurate within an error of less than 10% as compared to the measured strain data, indicating that the FRF prediction method is reliable enough to be used in practical applications.

The current treatment of choice for attention deficit hyperactivity disorder (ADHD) is pharmacotherapy. A search for new treatment options is underway, however, as the wide application of drugs to the general population of patients with ADHD is limited by side effects and the variance of pharmacokinetic effects of the drugs in each patient. In the present study, we applied repetitive transcranial magnetic stimulation (rTMS), a non-invasive treatment used in a number of other psychiatric disorders, to spontaneously hypertensive rats (SHRs), an animal model of ADHD, in order to assess the efficacy of the treatment in modifying behavioural symptoms as well as levels of dopamine, noradrenaline, serotonin, and brain-derived neurotrophic factor (BDNF). A total of fifteen sessions of high-frequency rTMS treatment were administered. Behavioural symptoms were observed using open field, Y-maze, and elevated plus-maze tests. Upon completion of the experiments, rats were sacrificed, and the neurochemical changes in brain tissue were analysed using high performance liquid chromatography and Western blotting. The SHRs treated with rTMS tended to exhibit less locomotor activity in the open field test over the course of treatment, but there was no improvement in inattention as measured by the Y-maze test. Furthermore, BDNF concentration increased and noradrenaline concentration decreased in the prefrontal cortex of SHRs treated with rTMS. The results of the present preclinical study indicate that rTMS may constitute a new modality of treatment for patients with ADHD, through further evaluation of specific treatment parameters as well as safety and efficacy in humans are required.

Yonghyun Lee,1,2,* Jungyun Kim,1,* Wooseong Kim,1,* Joon Nam,1,* Seongkeun Jeong,1 Sunyoung Lee,1 Jin-Wook Yoo,1 Min-Soo Kim,1 Yunjin Jung1 1College of Pharmacy, Pusan National University, Busan, 2Bio-Nanomedicine Laboratory, Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea *These authors contributed equally tothis work Abstract: Celecoxib, a selective cyclooxygenase-2 inhibitor, is potentially useful for the treatment of colonic diseases such as colorectal cancer and colitis. However, the cardiovascular toxicity of celecoxib limits its routine use in the clinic. Generally, colon-specific delivery of a drug both increases the therapeutic availability in the large intestine and decreases the systemic absorption of the drug, most likely resulting in enhanced therapeutic effects against colonic diseases such as colitis and reduced systemic side effects. To develop a colon-specific prodrug of celecoxib that could reduce its cardiovascular toxicity and improve its therapeutic activity, dextran–glutamic acid–celecoxib conjugate (glutam-1-yl celecoxib-dextran ester [G1CD]) was prepared and evaluated. While stable in pH 1.2 and 6.8 buffer solutions and small-intestinal contents, G1CD efficiently released celecoxib in cecal contents. Oral administration of G1CD to rats delivered a larger amount of celecoxib to the large intestine than free celecoxib. G1CD prevented the systemic absorption of celecoxib and did not decrease the serum level of 6-ketoprostaglandin F1α, an inverse indicator of cardiovascular toxicity of celecoxib. Collectively, G1CD may be a polymeric colon-specific celecoxib prodrug with therapeutic and toxicological advantages. Keywords: colon-specific drug delivery, dextran, celecoxib, prodrug, cardiovascular toxicity