An adaptable luminescence resonance energy transfer assay for measuring and screening protein-protein interactions and their inhibition

Protein-protein interactions (PPIs) are central to biological processes and represent an important class of therapeutic targets. Here we show that the interaction between FK506-binding protein 12 fused to green fluorescent protein (GFP-FKBP) and the rapamycin-binding domain of mTor fused to Escherichia coli dihydrofolate reductase (FRB-eDHFR) can be sensitively detected (signal-to-background ratio (S/B)>100) and accurately quantified within an impure cell lysate matrix using a luminescence resonance energy transfer (LRET) assay. Ascomycin-mediated inhibition of GFP-FKBP-rapamycin-FRB-eDHFR complex formation was also detected at high S/B ratio (>80) and Z'-factor (0.89). The method leverages the selective, stable binding of trimethoprim (TMP)-terbium complex conjugates to eDHFR, and time-resolved, background-free detection of the long-lifetime (～ms) terbium-to-GFP LRET signal that indicates target binding. TMP-eDHFR labeling can be adapted to develop high-throughput screening assays and complementary, quantitative counter-screens for a wide variety of PPI targets with a broad range of affinities that may not be amenable to purification.     

Topographic and biomechanical evaluation of cornea in patients with ichthyosis vulgaris

PurposeTo compare the topographic and biomechanical properties of corneas in eyes of patients with ichthyosis vulgaris (IV) and eyes of healthy individuals.MethodsThirty healthy individuals (control group) and 30 patients with IV (study group) were enrolled in this prospective study. Topographic measurements, including keratometry values, irregularity, and surface asymmetry index in the right eye of each participant were obtained using Scheimpflug camera with a Placido disc topographer (Sirius). Corneal hysteresis (CH), corneal resistance factor (CRF), corneal-compensated intraocular pressure (IOPcc) and Goldman-related intraocular pressure (IOPg) were measured using the Reichert Ocular Response Analyzer (ORA). Central corneal thickness (CCT) was also measured with ultrasonic pachymetry and the Sirius corneal topography system.ResultsTopographic parameters were not significantly different between both groups (p > 0.05). Although mean CH was not significantly different between the groups, the CRF was significantly lower in patients with IV (p = 0.249 and p = 0.005, respectively). The CCT was significantly lower in patients with IV compared to healthy controls (p < 0.001). The IOPg and IOPcc were significantly lower in the patients with ichthyosis than in healthy controls (p = 0.001 and p = 0.004, respectively).ConclusionsThe study demonstrated that while the eyes of patients with IV had corneal topographic findings and corneal hysteresis similar to those of healthy controls, some of corneal biomechanical properties such CRF and CCT and IOP values such as IOPg and IOPcc were significantly lower in patients with IV. These results should be taken into account when planning a corneal refractive surgery and glaucoma screening for patients with IV.     

Transient protein-protein interactions

Transient complexes are crucial for diverse biological processes such as biochemical pathways and signaling cascades in the cell. Here, we give an overview of the transient interactions; the importance of transient interactions as drug targets; and the structural characterization of transient protein-protein complexes based on the geometrical and physicochemical features of the transient complexes' interfaces. To better understand and eventually design transient protein-protein interactions (TPPIs), a molecular perspective of the protein-protein interfaces is necessary. Obtaining high-quality structures of protein-protein interactions could be one way of achieving this goal. After introducing the association kinetics of TPPIs, we elaborate on the experimental techniques detecting TPPIs in combination with the computational methods which classify transient and/or non-obligate complexes. In this review, currently available databases and servers that can be used to identify and predict TPPIs are also compiled.     

The effect of inappropriate dead time settings on the regulation of unevenness of material implemented by the automatic control: an application in the textile spinning industry

In this paper, we analyze the efficiency of regulation of unevenness material implemented by automatic control in case of inappropriate dead time settings. In this analysis, we consider various degrees of inaccurate dead time settings, different sizes, and occurring frequencies of the faults on the material. The results obtained indicate that in some cases, the regulation can still continue even in case of quite high degree of inaccurate dead time settings. However, the results obtained indicate also that the evenness of material after the regulation process can be worse than before regulation process in some cases. The work is carried out using simulation methodology and by considering the autoleveler equipment used as an automatic control system in the textile industry.     

水质自动监测记录仪的研制

指出了国内环保部门在水质监测方面存在的问题，阐述了研制水质自动监测仪的必要性。文中对该仪器的硬件和软件组成做了详细的介绍。同时对它的技术创新之处做了较详细的说明。     

Type-2 fuzzy model based controller design for neutralization processes

In this study, an inverse controller based on a type-2 fuzzy model control design strategy is introduced and this main controller is embedded within an internal model control structure. Then, the overall proposed control structure is implemented in a pH neutralization experimental setup. The inverse fuzzy control signal generation is handled as an optimization problem and solved at each sampling time in an online manner. Although, inverse fuzzy model controllers may produce perfect control in perfect model match case and/or non-existence of disturbances, this open loop control would not be sufficient in the case of modeling mismatches or disturbances. Therefore, an internal model control structure is proposed to compensate these errors in order to overcome this deficiency where the basic controller is an inverse type-2 fuzzy model. This feature improves the closed-loop performance to disturbance rejection as shown through the real-time control of the pH neutralization process. Experimental results demonstrate the superiority of the inverse type-2 fuzzy model controller structure compared to the inverse type-1 fuzzy model controller and conventional control structures.     

通用螺旋立铣刀的力学和动力学模型(二)

4模拟和试验结果 使用本文提出的广义方法,用各种刀具几何参数和材料,进行了超过300个切削试验.这里介绍螺旋和镶齿立铣实例,用以证明所提出模型的柔性.     

Development of a micro-punching machine and study on the influence of vibration machining in micro-EDM

This paper describes the development of a novel micro-punching machine that is capable of producing precision micro-holes. A significant feature of this machine is to fabricate the micro-punch and then the micro-die in the same machine, totally eliminating the eccentricity between the punch and the die when punching is proceeded. By applying vibration machining technique, we can decrease the possibility of electric short-circuiting during the micro-EDM process. The utilization of a proportional solenoid as the power unit of the micro-punching machine and as the source of vibration is found to be a successful attempt. Experiments to punch micro-holes with diameters of 0.1 and 0.2 mm on an SUS 304 stainless steel strip with 0.1 mm in thickness were carried out. The results show that the performance of this machine and the geometry of punched micro-holes are satisfactory.