Bead-based polymerase chain reaction on a microchip

We present a bead-based approach to microfluidic polymerase chain reaction (PCR), enabling fluorescent detection and sample conditioning in a single microchamber. Bead-based PCR, while not extensively investigated in microchip format, has been used in a variety of bioanalytical applications in recent years. We leverage the ability of bead-based PCR to accumulate fluorescent labels following DNA amplification to explore a novel DNA detection scheme on a microchip. The microchip uses an integrated microheater and temperature sensor for rapid control of thermal cycling temperatures, while the sample is held in a microchamber fabricated from (poly)dimethylsiloxane and coated with Parylene. The effects of key bead-based PCR parameters, including annealing temperature and concentration of microbeads in the reaction mixture, are studied to achieve optimized device sensitivity and detection time. The device is capable of detecting a synthetically prepared section of the Bordetella pertussis genome in as few as 10 temperature cycles with times as short as 15?min. We then demonstrate the use of the procedure in an integrated device; capturing, amplifying, detecting, and purifying template DNA in a single microfluidic chamber. These results show that this method is an effective method of DNA detection which is easily integrated in a microfluidic device to perform additional steps such as sample pre-conditioning.     

Ionic transport studies of the glassy silver vanadomolybdate system

Ionic conductivity of the Ag₂O-MoO₃-V₂0₅ system has been studied over a wide range of frequency, temperature and composition. A narrower glass forming region has been found in comparison to the corresponding Ag₂O-MoO₃-P₂O₅ and Ag₂O-B₂O₃-P₂O₅ systems. The highest conductivity at room temperature, _rt, = 3.21 × 10^–6–1 cm^–1 (d.c.) with an activation energy,E _act, of 0.466 eV, was observed for the glass former's ratio of unity. Further, it reached a maximum value of 2.2 × 10^–2¨-1 cm^–1 withE _act = 0.153 eV when the oxide-base glass was dissolved with Agl. D.c. conductivity, hopping rate and relaxation time in the present system have been found to be characterized by the same activation energy.     

基于光通信的数据单向导入系统的设计与实现

随着信息网络的日趋成熟,网络信息安全尤其是内网信息安全,越来越受到人们的关注,成为一时研究的热点。其中如何防止在移动存储设备与内网计算机信息交流过程中内网信息流失的问题,显得尤为迫切。为此,本文提出一种基于光通信的数据单向导入系统,为用户提供一种安全地将信息或者数据单向导入到内网的解决方案。利用光的单向性,从传输通道上卡断了反向的可能,有效地保护内网的机密信息不会流失到外部,同时做到良好的人机交互,用户可以任意选择存储设备中的文件发送。     

磁耦合谐振式无线能量传输的阻抗匹配特性

针对无线电能传输过程中传输效率随匹配阻抗大小变化的特点,提出两种阻抗匹配的方法:一种是基于步进电动机通过检测网络反馈信号,控制网络通过GA神经网络算法控制步进电动机,改变电容实现阻抗匹配;另一种是基于直直变换器中Cuk电路的占空比大小来匹配阻抗的方法.在负载和线圈之间嵌入单相桥式整流电路和Cuk升降压变压器实现匹配.根据最大传输功率理论,定量分析了改变电路的阻抗值大小,使系统的传输功率及效率最大.在MATLAB/Simulink平台验证了两种阻抗匹配设计方案的有效性.结果 表明,利用基于步进电动机和Cuk电路的阻抗匹配方法可以实现最佳阻抗值匹配,使无线电能传输效率最大化.     

Human DDX3X Unwinds Japanese Encephalitis and Zika Viral 5′ Terminal Regions

Flavivirus genus includes many deadly viruses such as the Japanese encephalitis virus (JEV) and Zika virus (ZIKV). The 5′ terminal regions (TR) of flaviviruses interact with human proteins and such interactions are critical for viral replication. One of the human proteins identified to interact with the 5′ TR of JEV is the DEAD-box helicase, DDX3X. In this study, we in vitro transcribed the 5′ TR of JEV and demonstrated its direct interaction with recombinant DDX3X (K_d of 1.66 ± 0.21 µM) using microscale thermophoresis (MST). Due to the proposed structural similarities of 5′ and 3′ TRs of flaviviruses, we investigated if the ZIKV 5′ TR could also interact with human DDX3X. Our MST studies suggested that DDX3X recognizes ZIKV 5′ TR with a K_d of 7.05 ± 0.75 µM. Next, we performed helicase assays that suggested that the binding of DDX3X leads to the unwinding of JEV and ZIKV 5′ TRs. Overall, our data indicate, for the first time, that DDX3X can directly bind and unwind in vitro transcribed flaviviral TRs. In summary, our work indicates that DDX3X could be further explored as a therapeutic target to inhibit Flaviviral replication     

Fluctuating NMDA Receptor Subunit Levels in Perirhinal Cortex Relate to Their Dynamic Roles in Object Memory Destabilization and Reconsolidation

Reminder cues can destabilize consolidated memories, rendering them modifiable before they return to a stable state through the process of reconsolidation. Older and stronger memories resist this process and require the presentation of reminders along with salient novel information in order to destabilize. Previously, we demonstrated in rats that novelty-induced object memory destabilization requires acetylcholine (ACh) activity at M₁ muscarinic receptors. Other research predominantly has focused on glutamate, which modulates fear memory destabilization and reconsolidation through GluN2B- and GluN2A-containing NMDARs, respectively. In the current study, we demonstrate the same dissociable roles of GluN2B- and N2A-containing NMDARs in perirhinal cortex (PRh) for object memory destabilization and reconsolidation when boundary conditions are absent. However, neither GluN2 receptor subtype was required for novelty-induced destabilization of remote, resistant memories. Furthermore, GluN2B and GluN2A subunit proteins were upregulated selectively in PRh 24 h after learning, but returned to baseline by 48 h, suggesting that NMDARs, unlike muscarinic receptors, have only a temporary role in object memory destabilization. Indeed, activation of M₁ receptors in PRh at the time of reactivation effectively destabilized remote memories despite inhibition of GluN2B-containing NMDARs. These findings suggest that cholinergic activity at M₁ receptors overrides boundary conditions to destabilize resistant memories when other established mechanisms are insufficient.     

基于无线网络的制造业物料配送系统的设计*

针对制造业传统物料配送系统存在的缺点,基于业务流程再造的理念,运用无线局域网技术设计了全新的物料配送系统。该系统能够对配送流程进行优化,提高了配送效率,有效地解决了配送不及时所造成的生产速度瓶颈问题。     

超导故障限流器抑制短路电流直流分量的仿真分析

短路电流中的周期分量和直流分量对系统短路容量都有贡献,而后者在工程应用中没有引起足够的重视.目前我国宁夏、上海、广州等电网普遍面临短路电流直流分量超标,系统保护装置难以可靠清除故障的状况,严重威胁电力系统的安全可靠性.基于此,本文提出使用超导故障限流器在限制短路电流幅值的同时,抑制短路电流直流分量的方法,缓解断路器的开断负担.目前超导限流器对短路电流直流分量的抑制效果及在高直流分量系统超导限流器的设计方法尚不清楚.因此,搭建了超导故障限流器模型,以330 kV系统为例,研究了不同直流分量时间常数下超导限流器阻值与直流分量的关系,提出了超导故障限流器失超阻值的优化设计方法.结果表明在不同的短路故障条件下,超导限流器对直流分量的抑制效果均非常明显,能有效提高系统的可靠性.     

An Engineered Biliverdin-Compatible Cyanobacteriochrome Enables a Unique Ultrafast Reversible Photoswitching Pathway

Cyanobacteriochromes (CBCRs) are promising optogenetic tools for their diverse absorption properties with a single compact cofactor-binding domain. We previously uncovered the ultrafast reversible photoswitching dynamics of a red/green photoreceptor AnPixJg2, which binds phycocyanobilin (PCB) that is unavailable in mammalian cells. Biliverdin (BV) is a mammalian cofactor with a similar structure to PCB but exhibits redder absorption. To improve the AnPixJg2 feasibility in mammalian applications, AnPixJg2_BV4 with only four mutations has been engineered to incorporate BV. Herein, we implemented femtosecond transient absorption (fs-TA) and ground state femtosecond stimulated Raman spectroscopy (GS-FSRS) to uncover transient electronic dynamics on molecular time scales and key structural motions responsible for the photoconversion of AnPixJg2_BV4 with PCB (Bpcb) and BV (Bbv) cofactors in comparison with the parent AnPixJg2 (Apcb). Bpcb adopts the same photoconversion scheme as Apcb, while BV4 mutations create a less bulky environment around the cofactor D ring that promotes a faster twist. The engineered Bbv employs a reversible clockwise/counterclockwise photoswitching that requires a two-step twist on ~5 and 35 picosecond (ps) time scales. The primary forward P_fr → P_o transition displays equal amplitude weights between the two processes before reaching a conical intersection. In contrast, the primary reverse P_o → P_fr transition shows a 2:1 weight ratio of the ~35 ps over 5 ps component, implying notable changes to the D-ring-twisting pathway. Moreover, we performed pre-resonance GS-FSRS and quantum calculations to identify the Bbv vibrational marker bands at ~659,797, and 1225 cm⁻¹. These modes reveal a stronger H-bonding network around the BV cofactor A ring with BV4 mutations, corroborating the D-ring-dominant reversible photoswitching pathway in the excited state. Implementation of BV4 mutations in other PCB-binding GAF domains like AnPixJg4, AM1_1870g3, and NpF2164g5 could promote similar efficient reversible photoswitching for more directional bioimaging and optogenetic applications, and inspire other bioengineering advances.     

The Effects of Insulin-Like Growth Factor I and BTP-2 on Acute Lung Injury

Acute lung injury (ALI) afflicts approximately 200,000 patients annually and has a 40% mortality rate. The COVID-19 pandemic has massively increased the rate of ALI incidence. The pathogenesis of ALI involves tissue damage from invading microbes and, in severe cases, the overexpression of inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). This study aimed to develop a therapy to normalize the excess production of inflammatory cytokines and promote tissue repair in the lipopolysaccharide (LPS)-induced ALI. Based on our previous studies, we tested the insulin-like growth factor I (IGF-I) and BTP-2 therapies. IGF-I was selected, because we and others have shown that elevated inflammatory cytokines suppress the expression of growth hormone receptors in the liver, leading to a decrease in the circulating IGF-I. IGF-I is a growth factor that increases vascular protection, enhances tissue repair, and decreases pro-inflammatory cytokines. It is also required to produce anti-inflammatory 1,25-dihydroxyvitamin D. BTP-2, an inhibitor of cytosolic calcium, was used to suppress the LPS-induced increase in cytosolic calcium, which otherwise leads to an increase in proinflammatory cytokines. We showed that LPS increased the expression of the primary inflammatory mediators such as toll like receptor-4 (TLR-4), IL-1β, interleukin-17 (IL-17), TNF-α, and interferon-γ (IFN-γ), which were normalized by the IGF-I + BTP-2 dual therapy in the lungs, along with improved vascular gene expression markers. The histologic lung injury score was markedly elevated by LPS and reduced to normal by the combination therapy. In conclusion, the LPS-induced increases in inflammatory cytokines, vascular injuries, and lung injuries were all improved by IGF-I + BTP-2 combination therapy.