Search

Abstract Nucleic acid sensing is involved in viral infections, immune response-related diseases, and therapeutics. Based on the composition of nucleic acids, nucleic acid sensors are defined as DNA or RNA sensors. Pathogen-associated nucleic acids are recognized by membrane-bound and intracellular receptors, known as pattern recognition receptors (PRRs), which induce innate immune-mediated antiviral responses. PRR activation is tightly regulated to eliminate infections and prevent abnormal or excessive immune responses. Nucleic acid sensing is an essential mechanism in tumor immunotherapy and gene therapies that target cancer and infectious diseases through genetically engineered immune cells or therapeutic nucleic acids. Nucleic acid sensing supports immune cells in priming desirable immune responses during tumor treatment. Recent studies have shown that nucleic acid sensing affects the efficiency of gene therapy by inhibiting translation. Suppression of innate immunity induced by nucleic acid sensing through small-molecule inhibitors, virus-derived proteins, and chemical modifications offers a potential therapeutic strategy. Herein, we review the mechanisms and regulation of nucleic acid sensing, specifically covering recent advances. Furthermore, we summarize and discuss recent research progress regarding the different effects of nucleic acid sensing on therapeutic efficacy. This study provides insights for the application of nucleic acid sensing in therapy.

Abstract Background Patients face a serious threat if a solid tumor leaves behind partial residuals or cannot be completely removed after surgical resection. Immunotherapy has attracted attention as a method to prevent this condition. However, the conventional immunotherapy method targeting solid tumors, that is, intravenous injection, has limitations in homing in on the tumor and in vivo expansion and has not shown effective clinical results. Method To overcome these limitations, NK cells (Natural killer cells) were encapsulated in micro/macropore-forming hydrogels using 3D bioprinting to target solid tumors. Sodium alginate and gelatin were used to prepare micro-macroporous hydrogels. The gelatin contained in the alginate hydrogel was removed because of the thermal sensitivity of the gelatin, which can generate interconnected micropores where the gelatin was released. Therefore, macropores can be formed through bioprinting and micropores can be formed using thermally sensitive gelatin to make macroporous hydrogels. Results It was confirmed that intentionally formed micropores could help NK cells to aggregate easily, which enhances cell viability, lysis activity, and cytokine release. Macropores can be formed using 3D bioprinting, which enables NK cells to receive the essential elements. We also characterized the functionality of NK 92 and zEGFR-CAR-NK cells in the pore-forming hydrogel. The antitumor effects on leukemia and solid tumors were investigated using an in vitro model. Conclusion We demonstrated that the hydrogel encapsulating NK cells created an appropriate micro–macro environment for clinical applications of NK cell therapy for both leukemia and solid tumors via 3D bioprinting. 3D bioprinting makes macro-scale clinical applications possible, and the automatic process shows potential for development as an off-the-shelf immunotherapy product. This immunotherapy system could provide a clinical option for preventing tumor relapse and metastasis after tumor resection. Graphical Abstract Micro/macropore-forming hydrogel with NK cells fabricated by 3D bioprinting and implanted into the tumor site.

Natural killer (NK) cells are immune effector cells with outstanding features for adoptive immunotherapy. Immune effector cells with chimeric antigen receptors (CARs) are promising targeted therapeutic agents for various diseases. Because tumor cells exhibit heterogeneous antigen expression and lose cell surface antigen expression during malignant progression, many CARs fixed against only one antigen have limited efficacy and are associated with tumor relapse. To expand the utility of CAR-NK cells, we designed a split and universal cotinine-CAR (Cot-CAR) system, comprising a Cot-conjugator and NK92 cells (α-Cot-NK92 cells) engineered with a CAR containing an anti-Cot-specific single-chain variable fragment and intracellular signaling domain. The efficacy of the Cot-CAR system was assessed in vitro using a cytolysis assay against various tumor cells, and its single- or multiple- utility potential was demonstrated using an in vivo lung metastasis model by injecting A549-Red-Fluc cells. The α-Cot-NK92 cells could switch targets, logically respond to multiple antigens, and tune cytolytic activation through the alteration of conjugators without re-engineering. Therefore the universal Cot-CAR system is useful for enhancing specificity and diversity of antigens, combating relapse, and controlling cytolytic activity. In conclusion, this universal Cot-CAR system reveals that multiple availability and controllability can be generated with a single, integrated system.

This paper proposes a reduction method for DC-link ripple current and common-mode voltage (CMV) in a hybrid active neutral-point-clamped (ANPC) inverter. A Si and SiC hybrid ANPC inverter has been developed recently to overcome the extremely high cost of a full-SiC ANPC inverter. A hybrid ANPC requires much fewer SiC MOSFETs than a full-SiC ANPC inverter while providing a comparable power density. Voltage source inverters such as hybrid ANPC inverters utilize electrolytic capacitors, which have a large capacitance per volume, as a DC link. However, an electrolytic capacitor is one of the most vulnerable components in a power electronic converter due to its small allowable ripple current. A large ripple current flowing into the electrolytic capacitor generates a heat loss, which shortens the lifetime of the capacitor. Furthermore, the common-mode voltage (CMV) causes an undesirable leakage current and electromagnetic interference. The CMV depends on the pulse-width modulation of the voltage source inverters. The proposed method enhances the reliability of the hybrid ANPC inverter by reducing the DC-link ripple current and CMV simultaneously. The effectiveness and validity of the proposed method are verified through simulations and experimental results.

A critical issue that needs to be addressed for wider utilization of hydrogen as fuel is protection against hydrogen embrittlement during cryogenic storage as it weakens the microstructure bonding force of metals through hydrogen penetration. Austenitic stainless steel, which is usually used in cryogenic vessels and is well known for its high hydrogen resistance at room temperature, has also been reported to be vulnerable to hydrogen embrittlement under cryogenic temperatures. In addition, because large storage vessels are operated over a wide range of temperatures, material behavior at various temperature conditions should also be considered. Therefore, in the present study, hydrogen charging of austenitic stainless steel was performed under various temperature conditions for carrying out prestrain and tensile tests. A decrease in the tensile strength and elongation and an increase in the yield strength were observed in all cases. In particular, the case of 20% prestrain at cryogenic temperature followed by tensile test at room temperature after hydrogen charging showed fracture in the elastic region. The hydrogen index was evaluated from the perspective of elongation and reduction in area, which are factors that indicate the degree of ductility. The aforementioned case showed the most severe results, while non-prestraining followed by tensile tests at room temperature was the least effected by hydrogen. In addition, the effect of strain-induced martensite on hydrogen embrittlement was analyzed using electron backscattered diffraction (EBSD). It was observed that the higher is the prestrain at cryogenic temperatures, the greater is the volume fraction of α’ martensite, which leads to hydrogen embrittlement. The edges and center of the fracture surface were analyzed using scanning electron microscopy (SEM). The hydrogen-charged specimens exhibited brittle fractures at the edges and ductile fractures at the center. The more severe the embrittlement, the more were the number of intergranular fractures and microdimples observed at the edges.

This paper proposes an improved discontinuous pulse-width modulation (DPWM) method based on phase-shifted PWM (PS-PWM) for three-phase cascaded H-bridge (CHB) multilevel inverters. Due to the CHB multilevel inverters employ a large number of power semiconductor switches, they suffer from device failures because the switches are one of the major failure factors in a power electronic system. Therefore, the reliability and life-time of CHB multilevel inverters are mainly determined by the reliability of switches. The most failure of switches is occurred by its accumulated thermal stress which is generated by power losses including the conduction and switching loss. By applying the conventional DPWM, the life-time of switches can be extended because it reduces the switching loss. The conventional DPWM, however, still operates with high speed carrier frequency and it is not enough to reduce the switching loss of the CHB multilevel inverter with a large number of inverter cells. The proposed novel DPWM method provides the improved switching loss reduction through the different switching behaviors for each leg. Additionally, the rotation scheme promises even power distribution and life-time management among inverter cells in a CHB multilevel inverter. The simulation results and experimental results confirm the switching loss reduction and even power distribution abilities of the proposed modulation method.

In this paper, an optimized space-vector modulation (SVM) to reduce DC-link ripple current in three-level inverters is presented. Various types of capacitors can be utilized to balance the voltage in the DC-link of voltage source inverters. Electrolytic capacitors are widely used owing to their large capacitance per volume. However, electrolytic capacitors have a short lifespan because the allowable ripple current is low. A conventional SVM that is focused on improving harmonic characteristics. Therefore, it cannot balance the DC-link ripple current properly because using some voltage vectors increases the DC-link ripple current. To overcome this limitation and extend the lifespan of the capacitors, an optimized SVM is proposed to reduce the DC-link ripple current. The proposed modulation scheme synthesizes the reference voltage by choosing voltage vectors that cause smaller increases in the DC-link ripple current, avoiding those that cause larger increases. The effectiveness of the proposed optimized SVM is verified by simulations and experimental results.

This paper proposes open fault diagnosis and tolerance control methods for grid-connected hybrid active neutral-point-clamped (ANPC) inverters with optimized carrier-based pulse width modulation. The hybrid ANPC inverter is composed of silicon carbide metal-oxide-semiconductor field effect transistors and Si-insulated gate bipolar transistors. Open faults of switch devices lead to the distortion of output currents, inverter performance degradation, and low reliability. Therefore, the open-circuit fault diagnosis and the tolerance control methods should be handled to improve the reliability of the inverter. The fault tolerance control methods for grid-connected inverters should also consider the fixed voltage modulation index as the modulation index is fixed. In this study, the proposed fault tolerance control method handles the open fault without reducing the modulation index. The implementation of the proposed method is easy and can be conducted through a simple change in the reference waveform. The effectiveness of the proposed fault detection and fault tolerance control methods was verified through the simulation and experimental results.

The cascaded H-bridge multilevel inverter (CHMI) is a modular structure that consists of many power semiconductor switches. With this increase in the number of power semiconductor switches, it is hard to predict and handle the failure of the devices, and hence reliability of CHMI decreases. The major cause of power semiconductor switch failure is junction temperature that is produced by power losses. The study proposes a multi-carrier pulse-width modulation (PWM) scheme for reduction in switching losses of CHMI. In the proposed modulation scheme, the two legs conduct switching operation at different frequencies for switching reduction. One leg conducts switching operation with high frequency, while the other leg conducts switching operation with fundamental frequency. The switching operations with different frequencies cause unbalanced switching loss to each leg. Therefore, the junction temperature that is based on power losses leads to different life-times for the power semiconductor switch. Additionally, the switching frequency of the two legs is alternated to evenly distribute switching losses and junction temperature. Simulation and experimental results verify the performance of the proposed PWM scheme.

Abstract We investigate the thermal stability of alumina supporting layers sputtered at different conditions and its effect on the growth of aligned single-walled carbon nanotube arrays. Radio frequency magnetron sputtering of alumina under oxygen–argon atmosphere produces a Si-rich alumina alloy film on a silicon substrate. Atomic force microscopy on the annealed catalysts reveals that Si-rich alumina films are more stable than alumina layers with low Si content at the elevated temperatures at which the growth of single-walled carbon nanotubes is initiated. The enhanced thermal stability of the Si-rich alumina layer results in a narrower (

MicroRNA (miR)-150 is a developmental regulator of several immune-cell types, but its role in CD8+ T cells is largely unexplored. Here, we show that miR-150 regulates the generation of memory CD8+ T cells. After acute virus infection, miR-150 knockout (KO) mice exhibited an accelerated differentiation of CD8+ T cells into memory cells and improved production of effector cytokines. Additionally, miR-150 KO CD8+ T cells displayed an enhanced recall response and improved protection against infections with another virus and bacteria. We found that forkhead box O1 (Foxo1) and T cell-specific transcription factor 1 (TCF1) are upregulated during the early activation phase in miR-150 KO CD8+ T cells and that miR-150 directly targets and suppresses Foxo1. These results suggest that miR-150-mediated suppression of Foxo1 regulates the balance between effector and memory cell differentiation, which might aid in the development of improved vaccines and T cell therapeutics.

Micro/nano-precision glass molding (MNPGM) is an efficient approach for manufacturing micro/nanostructured glass components with intricate geometry and a high-quality optical finish. In MNPGM, the mold, which directly imprints the desired pattern on the glass substrate, is a key component. To date, a wide variety of mold inserts have been utilized in MNPGM. The aim of this article is to review the latest advances in molds for MNPGM and their fabrication methods. Surface finishing is specifically addressed because molded glass is usually intended for optical applications in which the surface roughness should be lower than the wavelength of incident light to avoid scattering loss. The use of molds for a wide range of molding temperatures is also discussed in detail. Finally, a series of tables summarizing the mold fabrication methods, mold patterns and their dimensions, anti-adhesion coatings, molding conditions, molding methods, surface roughness values, glass substrates and their glass transition temperatures, and associated applications are presented. This review is intended as a roadmap for those interested in the glass molding field.

The design or dimension of micro-supercapacitor electrodes is an important factor that determines their performance. In this study, a microsupercapacitor was precisely fabricated on a silicon substrate by irradiating an imprinted furan micropattern with a CO2 laser beam under ambient conditions. Since furan is a carbon-abundant polymer, electrically conductive and porous carbon structures were produced by laser-induced pyrolysis. While the pyrolysis of a furan film in a general electric furnace resulted in severe cracks and delamination, the laser pyrolysis method proposed herein yielded porous carbon films without cracks or delamination. Moreover, as the imprinting process already designated the furan area for laser pyrolysis, high-precision patterning was achieved in the subsequent laser pyrolysis step. This two-step process exploited the superior resolution of imprinting for the fabrication of a laser-pyrolyzed carbon micropattern. As a result, the technical limitations of conventional laser direct writing could be overcome. The laser-pyrolyzed carbon structure was employed for microsupercapacitor electrodes. The microsupercapacitor showed a specific capacitance of 0.92 mF/cm2 at 1 mA/cm2 with a PVA-H2SO4 gel electrolyte, and retained an up to 88% capacitance after 10,000 charging/discharging cycles.

Although polymer nanoimprinting on glass substrates has been widely employed for the fabrication of functional anti-reflective (AR) nanostructures, several drawbacks exist with respect to durability and delamination. The direct patterning of glass material is a potential solution for outdoor applications that require AR functional nanostructured glass plates. In this study, a glass imprinting technique was employed for the fabrication of an AR nanostructure on a soda-lime glass substrate using a vitreous carbon (VC) stamp. The VC stamp, which had a high aspect ratio nanopost array with a pitch of 325 nm, diameter of 110 nm, and height of ~220 nm, was fabricated by the carbonization of a replicated Furan precursor from an Si master. During the glass imprinting process using the nanopost array VC stamp, the softened glass material gradually protruded into the spaces between the nanopins owing to viscoelastic behavior, and one can achieve a cross-sinusoidal surface relief under specific imprinting condition, which can be used as an AR nanostructure with a gradually increasing refractive index. The effects of the processing temperature on the surface profile of the glass imprinted parts and the measured transmission spectra were analyzed, and a glass imprinting temperature of 700 °C and pressure of 1 MPa were found to be the optimum condition. The height of the fabricated cross-sinusoidal nanostructure was 80 nm, and the light transmission was increased by ~2% over the entire visible-light range. Furthermore, the measured transmission spectrum observed to be in good agreement with the simulation results.

The effects of structural design parameters on the performance of nano-replicated photonic crystal (PC) label-free biosensors were examined by the analysis of simulated reflection spectra of PC structures. The grating pitch, duty, scaled grating height and scaled TiO2 layer thickness were selected as the design factors to optimize the PC structure. The peak wavelength value (PWV), full width at half maximum of the peak, figure of merit for the bulk and surface sensitivities, and surface/bulk sensitivity ratio were also selected as the responses to optimize the PC label-free biosensor performance. A parametric study showed that the grating pitch was the dominant factor for PWV, and that it had low interaction effects with other scaled design factors. Therefore, we can isolate the effect of grating pitch using scaled design factors. For the design of PC-label free biosensor, one should consider that: (1) the PWV can be measured by the reflection peak measurement instruments, (2) the grating pitch and duty can be manufactured using conventional lithography systems, and (3) the optimum design is less sensitive to the grating height and TiO2 layer thickness variations in the fabrication process. In this paper, we suggested a design guide for highly sensitive PC biosensor in which one select the grating pitch and duty based on the limitations of the lithography and measurement system, and conduct a multi objective optimization of the grating height and TiO2 layer thickness for maximizing performance and minimizing the influence of parameter variation. Through multi-objective optimization of a PC structure with a fixed grating height of 550 nm and a duty of 50%, we obtained a surface FOM of 66.18 RIU−1 and an S/B ratio of 34.8%, with a grating height of 117 nm and TiO2 height of 210 nm.

The detection of body fluids has been used to identify a suspect and build a criminal case. As the amount of evidence collected at a crime site is limited, a multiplex identification system for body fluids using a small amount of sample is required. In this study, we proposed a multiplex detection platform using an Ag vertical nanorod metal enhanced fluorescence (MEF) substrate for semen and vaginal fluid (VF), which are important evidence in cases of sexual crime. The Ag nanorod MEF substrate with a length of 500 nm was fabricated by glancing angle deposition, and amino functionalization was conducted to improve binding ability. The effect of incubation time was analyzed, and an incubation time of 60 min was selected, at which the fluorescence signal was saturated. To assess the performance of the developed identification chip, the identification of semen and VF was carried out. The developed sensor could selectively identify semen and VF without any cross-reactivity. The limit of detection of the fabricated microarray chip was 10 times better than the commercially available rapid stain identification (RSID) Semen kit.

We report on the study of energy-harvesting performance in medium-size (400 cm2) glass-based semitransparent solar concentrators employing edge-mounted photovoltaic modules. Systems using several different types of glazing system architecture and containing embedded diffractive structures are prepared and characterized. The technological approaches to the rapid manufacture of large-area diffractive elements suitable for use in solar window-type concentrators are described. These elements enable the internal deflection and partial trapping of light inside glass-based concentrator windows. We focus on uncovering the potential of pattern-transfer polymer-based soft lithography for enabling both the improved photon collection probability at solar cell surfaces, and the up-scaling of semitransparent solar window dimensions. Results of photovoltaic characterization of several solar concentrators employing different internal glazing-system structure and diffractive elements produced using different technologies are reported and discussed.

The application of microtechnology to traditional mechanical industries is limited owing to the lack of suitable micropatterning technology for durable materials including metal. In this research, a glassy carbon (GC) micromold was applied for the direct metal forming (DMF) of a microstructure on an aluminum (Al) substrate. The GC mold with microdome cavities was prepared by carbonization of a furan precursor, which was replicated from the thermal reflow photoresist master pattern. A microdome array with a diameter of 8.4 μm, a height of ~0.74 μm, and a pitch of 9.9 μm was successfully fabricated on an Al substrate by using DMF at a forming temperature of 645 °C and an applied pressure of 2 MPa. As a practical application of the proposed DMF process, the enhanced boiling heat transfer characteristics of the DMF microdome Al substrate were analyzed. The DMF microdome Al substrate showed 20.4 ± 2.6% higher critical heat flux and 34.1 ± 5.3% higher heat transfer coefficient than those of a bare Al substrate.

This paper presents a method to improve the quality of input-output currents in a three-level neutral-point clamped (NPC) inverter with small direct current-link (DC-link) capacitor systems. The inverter systems with the small DC-link capacitors have several advantages in terms of cost, volume, life-time, and reliability when compared to inverters that use large DC-link capacitors. However, there are problems with respect to the deterioration of the input current quality and a severe ripple of neutral-point voltage (NPV), which can cause an aggravated output current. To mitigate these issues, an additional circuit is applied for the input current shaping and a compensation algorithm is applied to reduce the ripple voltage of NPV. The effectiveness of the proposed design and control method is verified with various simulation and experimental results.

This study reports a cost-effective method of replicating glass microfluidic chips using a vitreous carbon (VC) stamp. A glass replica with the required microfluidic microstructures was synthesized without etching. The replication method uses a VC stamp fabricated by combining thermal replication using a furan-based, thermally-curable polymer with carbonization. To test the feasibility of this method, a flow focusing droplet generator with flow-focusing and channel widths of 50 µm and 100 µm, respectively, was successfully fabricated in a soda-lime glass substrate. Deviation between the geometries of the initial shape and the vitreous carbon mold occurred because of shrinkage during the carbonization process, however this effect could be predicted and compensated for. Finally, the monodispersity of the droplets generated by the fabricated microfluidic device was evaluated.

An all-glass bifurcation microfluidic chip for blood plasma separation was fabricated by a cost-effective glass molding process using an amorphous carbon (AC) mold, which in turn was fabricated by the carbonization of a replicated furan precursor. To compensate for the shrinkage during AC mold fabrication, an enlarged photoresist pattern master was designed, and an AC mold with a dimensional error of 2.9% was achieved; the dimensional error of the master pattern was 1.6%. In the glass molding process, a glass microchannel plate with negligible shape errors (~1.5%) compared to AC mold was replicated. Finally, an all-glass bifurcation microfluidic chip was realized by micro drilling and thermal fusion bonding processes. A separation efficiency of 74% was obtained using the fabricated all-glass bifurcation microfluidic chip.