DNA Nanotechnology for Investigating Mechanical Signaling in the Immune System

Immune recognition occurs at specialized cell-cell junctions when immune cells and target cells physically touch. In this junction, groups of receptor-ligand complexes assemble and experience molecular forces that are ultimately generated by the cellular cytoskeleton. These forces are in the range of piconewton (pN) but play crucial roles in immune cell activation and subsequent effector responses. In this minireview, we will review the development of DNA based molecular tension sensors and their applications in mapping and quantifying mechanical forces experienced by immunoreceptors including T-cell receptor (TCR), Lymphocyte function-associated antigen (LFA-1), and the B-cell receptor (BCR) among others. In addition, we will highlight the use of DNA as a mechanical gate to manipulate mechanotransduction and decipher how mechanical forces regulate antigen discrimination and receptor signaling.     

Proteomics-Based Discovery of First-in-Class Chemical Probes for Programmed Cell Death Protein 2 (PDCD2)

Chemical probes are essential tools for understanding biological systems and for credentialing potential biomedical targets. Programmed cell death 2 (PDCD2) is a member of the B-cell lymphoma 2 (Bcl-2) family of proteins, which are critical regulators of apoptosis. Here we report the discovery and characterization of 10 e , a first-in-class small molecule degrader of PDCD2. We discovered this PDCD2 degrader by serendipity using a chemical proteomics approach, in contrast to the conventional approach for making bivalent degraders starting from a known binding ligand targeting the protein of interest. Using 10 e as a pharmacological probe, we demonstrate that PDCD2 functions as a critical regulator of cell growth by modulating the progression of the cell cycle in T lymphoblasts. Our work provides a useful pharmacological probe for investigating PDCD2 function and highlights the use of chemical proteomics to discover selective small molecule degraders of unanticipated targets.     

Detection of Unfolded Cellular Proteins Using Nanochannel Arrays with Probe-Functionalized Outer Surfaces

Nanochannel technology has emerged as a powerful tool for label-free and highly sensitive detection of protein folding/unfolding status. However, utilizing the inner walls of a nanochannel array may cause multiple events even for proteins with the same conformation, posing challenges for accurate identification. Herein, we present a platform to detect unfolded proteins through electrical and optical signals using nanochannel arrays with outer-surface probes. The detection principle relies on the specific binding between the maleimide groups in outer-surface probes and the protein cysteine thiols that induce changes in the ionic current and fluorescence intensity responses of the nanochannel array. By taking advantage of this mechanism, the platform has the ability to differentiate folded and unfolded state of proteins based on the exposure of a single cysteine thiol group. The integration of these two signals enhances the reliability and sensitivity of the identification of unfolded protein states and enables the distinction between normal cells and Huntington's disease mutant cells. This study provides an effective approach for the precise analysis of proteins with distinct conformations and holds promise for facilitating the diagnoses of protein conformation-related diseases.     

Near-Infrared Photodynamic Chemiluminescent Probes for Cancer Therapy and Metastasis Detection

There is growing interest in the development of chemiluminescence (CL) probes for phototheranostics because of their minimized tissue autofluorescence. However, due to a lack of near-infrared (NIR)-absorbing chemiluminophores, current probes for NIR CL-guided phototherapy are based on nanoparticles made up of multiple components. We report bright unimolecular chemiluminophores with NIR absorptions and emissions, long CL half-lives and ideal photodynamic efficiency. One luminophore is modified into an activatable probe, DBP_OL, with a turn-on CL signal and photodynamic activity that are specific to a cancer biomarker. The highly sensitive DBP_OL allows CL-guided photodynamic therapy which completely inhibits tumor growth and lung metastasis in mouse models, and can be applied for noninvasive monitoring of lung metastasis. We provide molecular guidelines for NIR-absorbing CL probes for imaging-guided phototherapy.     

比率型荧光探针的设计合成及其对Al3+识别性能研究

为了进一步提高金属离子探针的选择性和灵敏性,以3,5-二氨基苯甲酸和2,4-二羟基苯甲醛为原料,设计合成了一种比率型金属离子探针L(3,5-二(2,4-二羟基）苯基亚氨基)苯甲酸)。通过核磁、质谱和红外等方法对探针L的结构进行了表征,并利用荧光光谱、紫外光谱对探针L的光学性能进行了研究,结果表明：L对Al3+具有良好的选择性和灵敏性,受其它金属离子的干扰较小,并且反应前后伴有明显的颜色变化（黄色到无色）。通过计算,缔合常数为Kb = 1.35×104,L对Al3+的检出限低至8.6×10-8 mol/L,低于饮用水和食品中铝含量的国家标准,该离子探针有望应用于食品和水环境中Al3+的检测。     

铜Ⅱ和水合肼双功能螺吡喃探针的合成及应用

以4-甲基-2,6-二甲酰基苯酚和1,3,3-三甲基-2-亚甲基吲哚啉为原料合成了能同时识别Cu2＋和水合肼的螺吡喃双功能探针L,其结构经1 H NMR、13 C NMR、FT-IR和 H RMS进行了表征。通过紫外-可见光谱法、荧光光谱法、核磁共振和质谱研究了探针L在pH=7.40的Trus-HCl-乙醇溶液(1∶1, V/V)中与19种金属离子、18种阴离子及9种胺类化合物的识别特性。结果表明,在pH=7.40的Trus-HCl-乙醇溶液(1∶1, V/V)中,相对于其它干扰离子,L对Cu2＋和水合肼具有特异选择性和专一性,制成试纸能够实现肉眼识别检测含μmol/L Cu2＋的水样。尤为重要的是,制备的含L的TLC板能实现在紫外灯下对液体和气体水合肼的肉眼识别。将L应用于水样和药物中Cu2＋或水合肼测定的回收率在83.5%~111.0%之间,相对标准偏差( RSD)<4%,说明L适用于环境污染监测和药物分析检测,具有潜在的应用前景。     

Ultrasonication an intensifying tool for preparation of stable nanofluids and study the time influence on distinct properties of graphene nanofluids – A systematic overview

Optimum ultrasonication time will lead to the better performance for heat transfer in addition to preparation methods and thermal properties of the nanofluids. Nano particles are dispersed in base fluids like water (water-based fluids), glycols (glycol base fluids) &oils at different mass or volume fraction by using different preparation techniques. Significant preparation technique can enhance the stability, effects various parameters & thermo-physical properties of fluids. Agglomeration of the dispersed nano particles will lead to declined thermal performance, thermal conductivity, and viscosity. For better dispersion and breaking down the clusters, Ultrasonication method is the highly influential approach. Sonication hour is unique for different nano fluids depending on their response to several considerations. In this review, systematic investigations showing effect on various physical and thermal properties based on ultrasonication/ sonication time are illustrated. In this analysis it is found that increased power or time of ideal sonication increases the dispersion, leading to higher stable fluids, decreased particle size, higher thermal conductivity, and lower viscosity values. Employing the ultrasonic probe is substantially more effective than ultrasonic bath devices. Low ultrasonication power and time provides best outcome. Various sonication time periods by various research are summarized with respect to the different thermophysical properties. This is first review explaining sonication period influence on thermophysical properties of graphene nanofluids.     

Azo-pyrene–based fluorescent sensor of reductive cleavage of isomeric azo functional group

In this study we investigated the reductive azo cleavage of an azo compound presenting a pyrene fluorophore (Azo-py). Because of dramatic changes in its fluorescence, Azo-py could be used as a monitoring system for the reductive azo cleavage. Electron transfer from the pyrene unit to the azo moiety induced fluorescence quenching; this quenched fluorescence was recovered after the reductive azo cleavage. IR and NMR spectroscopy were used to study the various structural states. The rate of reductive cleavage of the azo compound, determined through fluorescence monitoring, depended on its structural state: the cleavage of trans-Azo-py was much faster than that of the cis-Azo-py. Furthermore, the Azo-py fluorophore was highly sensitive to the presence of zinc, but not other metal compounds, and the pH.     

An efficient (1)H/(31)P double-resonance solid-state NMR probe that utilizes a scroll coil

The construction and performance of a scroll coil double-resonance probe for solid-state NMR on stationary samples is described. The advantages of the scroll coil at the high resonance frequencies of (1)H and (31)P include: high efficiency, minimal perturbations of tuning by a wide range of samples, minimal RF sample heating of high dielectric samples of biopolymers in aqueous solution, and excellent RF homogeneity. The incorporation of a cable tie cinch for mechanical stability of the scroll coil is described. Experimental results obtained on a Hunter Killer Peptide 1 (HKP1) interacting with phospholipid bilayers of varying lipid composition demonstrate the capabilities of this probe on lossy aqueous samples.     

Software note: using probe secondary structure information to enhance Affymetrix GeneChip background estimates

High-density short oligonucleotide microarrays are a primary research tool for assessing global gene expression. Background noise on microarrays comprises a significant portion of the measured raw data. A number of statistical techniques have been developed to correct for this background noise. Here, we demonstrate that probe minimum folding energy and structure can be used to enhance a previously existing model for background noise correction. We estimate that probe secondary structure accounts for up to 3% of all variation on Affymetrix microarrays.