Visualizing Oxidative Cellular Stress Induced by Nanoparticles in the Subcytotoxic Range Using Fluorescence Lifetime Imaging

Nanoparticles hold a great promise in biomedical science. However, due to their unique physical and chemical properties they can lead to overproduction of intracellular reactive oxygen species (ROS). As an important mechanism of nanotoxicity, there is a great need for sensitive and high‐throughput adaptable single‐cell ROS detection methods. Here, fluorescence lifetime imaging microscopy (FLIM) is employed for single‐cell ROS detection (FLIM‐ROX) providing increased sensitivity and enabling high‐throughput analysis in fixed and live cells. FLIM‐ROX owes its sensitivity to the discrimination of autofluorescence from the unique fluorescence lifetime of the ROS reporter dye. The effect of subcytotoxic amounts of cationic gold nanoparticles in J774A.1 cells and primary human macrophages on ROS generation is investigated. FLIM‐ROX measures very low ROS levels upon gold nanoparticle exposure, which is undetectable by the conventional method. It is demonstrated that cellular morphology changes, elevated senescence, and DNA damage link the resulting low‐level oxidative stress to cellular adverse effects and thus nanotoxicity. Multiphoton FLIM‐ROX enables the quantification of spatial ROS distribution in vivo, which is shown for skin tissue as a target for nanoparticle exposure. Thus, this innovative method allows identifying of low‐level ROS in vitro and in vivo and, subsequently, promotes understanding of ROS‐associated nanotoxicity.     

Blood and Oxidative Stress (BOS): Soyuz mission “Eneide”

The aim of this experiment was to assay astronaut antioxidant status, to analyse red blood cell membrane composition of astronauts prior and after flight and to study the correlation with oxidative stress that erythrocytes have undergone due to space radiations. Results obtained from this single case study, indicate that during a short time flight, erythrocytes decrease their antioxidant defences, to counteract oxidative stress.     

Understanding the Antibacterial Mechanism of CuO Nanoparticles: Revealing the Route of Induced Oxidative Stress

To date, there is still a lack of definite knowledge regarding the interaction of CuO nanoparticles with bacteria and the possible permeation of the nanoparticles into bacterial cells. This study was aimed at shedding light on the size‐dependent (from the microscale down to the small nanoscale) antibacterial activity of CuO. The potent antibacterial activity of CuO nanoparticles was found to be due to ROS‐generation by the nanoparticles attached to the bacterial cells, which in turn provoked an enhancement of the intracellular oxidative stress. This paradigm was confirmed by several assays such as lipid peroxidation and reporter strains of oxidative stress. Furthermore, electron microscopy indicated that the small nanoparticles of CuO penetrated the cells. Collectively, the results reported herein may reconcile conflicting concepts in the literature concerning the antibacterial mechanism of CuO nanoparticles, as well as highlight the potential for developing sustainable CuO nanoparticles‐based devices for inhibiting bacterial infections.     

Synthesis of a Potential Tumor Imaging Agent by Oxidative Radioiodination of Aspirin and Its Preclinical Study

Radiochemistry - Aspirin, one of nonsteroidal anti-inflammatory analgesic drugs, was labeled with 125I with a labeling yield of 85.5% under the following conditions: pH 9, 100 mg of the substrate,...     

Gold Nanoparticles of Diameter 1.4 nm Trigger Necrosis by Oxidative Stress and Mitochondrial Damage

Gold nanoparticles (AuNPs) are generally considered nontoxic, similar to bulk gold, which is inert and biocompatible. AuNPs of diameter 1.4 nm capped with triphenylphosphine monosulfonate (TPPMS), Au1.4MS, are much more cytotoxic than 15‐nm nanoparticles (Au15MS) of similar chemical composition. Here, major cell‐death pathways are studied and it is determined that the cytotoxicity is caused by oxidative stress. Indicators of oxidative stress, reactive oxygen species (ROS), mitochondrial potential and integrity, and mitochondrial substrate reduction are all compromised. Genome‐wide expression profiling using DNA gene arrays indicates robust upregulation of stress‐related genes after 6 and 12 h of incubation with a 2 × IC50 concentration of Au1.4MS but not with Au15MS nanoparticles. The caspase inhibitor Z‐VAD‐fmk does not rescue the cells, which suggests that necrosis, not apoptosis, is the predominant pathway at this concentration. Pretreatment of the nanoparticles with reducing agents/antioxidants N‐acetylcysteine, glutathione, and TPPMS reduces the toxicity of Au1.4MS. AuNPs of similar size but capped with glutathione (Au1.1GSH) likewise do not induce oxidative stress. Besides the size dependency of AuNP toxicity, ligand chemistry is a critical parameter determining the degree of cytotoxicity. AuNP exposure most likely causes oxidative stress that is amplified by mitochondrial damage. Au1.4MS nanoparticle cytotoxicity is associated with oxidative stress, endogenous ROS production, and depletion of the intracellular antioxidant pool.     

Tailoring Enzyme‐Like Activities of Gold Nanoclusters by Polymeric Tertiary Amines for Protecting Neurons Against Oxidative Stress

The cytotoxicity of nanozymes has drawn much attention recently because their peroxidase‐like activity can decompose hydrogen peroxide (H₂O₂) to produce highly toxic hydroxyl radicals (?OH) under acidic conditions. Although catalytic activities of nanozymes are highly associated with their surface properties, little is known about the mechanism underlying the surface coating‐mediated enzyme‐like activities. Herein, it is reported for the first time that amine‐terminated PAMAM dendrimer‐entrapped gold nanoclusters (AuNCs‐NH₂) unexpectedly lose their peroxidase‐like activity while still retaining their catalase‐like activity in physiological conditions. Surprisingly, the methylated form of AuNCs‐NH₂ (i.e., MAuNCs‐N⁺R₃, where R = H or CH₃) results in a dramatic recovery of the intrinsic peroxidase‐like activity while blocking most primary and tertiary amines (1°‐ and 3°‐amines) of dendrimers to form quaternary ammonium ions (4°‐amines). However, the hidden peroxidase‐like activity is also found in hydroxyl‐terminated dendrimer‐encapsulated AuNCs (AuNCs‐OH, inside backbone with 3°‐amines), indicating that 3°‐amines are dominant in mediating the peroxidase‐like activity. The possible mechanism is further confirmed that the enrichment of polymeric 3°‐amines on the surface of dendrimer‐encapsulated AuNCs provides sufficient suppression of the critical mediator ?OH for the peroxidase‐like activity. Finally, it is demonstrated that AuNCs‐NH₂ with diminished cytotoxicity have great potential for use in primary neuronal protection against oxidative damage.     

Massively Evoking Immunogenic Cell Death by Focused Mitochondrial Oxidative Stress using an AIE Luminogen with a Twisted Molecular Structure

Immunogenic cell death (ICD) provides momentous theoretical principle for modern cancer immunotherapy. However, the currently available ICD inducers are still very limited and photosensitizer‐based ones can hardly induce sufficient ICD to achieve satisfactory cancer immunotherapy by themselves. Herein, an organic photosensitizer (named TPE‐DPA‐TCyP) with a twisted molecular structure, strong aggregation‐induced emission activity, and specific ability is reported for effectively inducing focused mitochondrial oxidative stress of cancer cells, which can serve as a much superior ICD inducer to the popularly used ones, including chlorin e6 (Ce6), pheophorbide A, and oxaliplatin. Furthermore, more effective in vivo ICD immunogenicity of TPE‐DPA‐TCyP than Ce6 is also demonstrated using a prophylactic tumor vaccination model. The underlying mechanism of the effectiveness and robustness of TPE‐DPA‐TCyP in inducing antitumor immunity and immune‐memory effect in vivo is verified by immune cell analyses. This study thus reveals that inducing focused mitochondrial oxidative stress is a highly effective strategy to evoke abundant and large‐scale ICD.     

Physical Forces Modulate Oxidative Status and Stress Defense Meditated Metabolic Adaptation of Yeast Colonies: Spaceflight and Microgravity Simulations

Baker’s yeast (Saccharomyces cerevisiae) has broad genetic homology to human cells. Although typically grown as 1-2mm diameter colonies under certain conditions yeast can form very large (10 + mm in diameter) or ‘giant’ colonies on agar. Giant yeast colonies have been used to study diverse biomedical processes such as cell survival, aging, and the response to cancer pharmacogenomics. Such colonies evolve dynamically into complex stratified structures that respond differentially to environmental cues. Ammonia production, gravity driven ammonia convection, and shear defense responses are key differentiation signals for cell death and reactive oxygen system pathways in these colonies. The response to these signals can be modulated by experimental interventions such as agar composition, gene deletion and application of pharmaceuticals. In this study we used physical factors including colony rotation and microgravity to modify ammonia convection and shear stress as environmental cues and observed differences in the responses of both ammonia dependent and stress response dependent pathways We found that the effects of random positioning are distinct from rotation. Furthermore, both true and simulated microgravity exacerbated both cellular redox responses and apoptosis. These changes were largely shear-response dependent but each model had a unique response signature as measured by shear stress genes and the promoter set which regulates them These physical techniques permitted a graded manipulation of both convection and ammonia signaling and are primed to substantially contribute to our understanding of the mechanisms of drug action, cell aging, and colony differentiation.     

Oxidative stabilization of acrylic fibres

A new model for the structure of oriented acrylic fibres is presented. The polyacrylonitrile molecules (or the acrylic sequences in a co-polymer) are suggested to form two distinct regions within a fibril: amorphous (disordered) and partially ordered. In the partially ordered regions, the polymer molecules assume a contorted helical shape to form rods with a diameter averaging about 6.0 Å in which the nitrile units are oriented at various angles to the rod axis, but are spaced irregularly on or near the surface of the rod. The nitrile groups of adjacent rods can interpenetrate to form dipole pairs. The rods are ordered into a liquid crystal-type array, giving in some cases a lamellar-like texture oriented perpendicular to the fibril axis, with the ordered lamellae regions interspersed with amorphous regions. Evidence for the structure is obtained from transmission electron microscopy observations, a transient peak observed in small-angle X-ray scattering when fibres are thermally treated, as well as wide-angle X-ray diffraction patterns. The proposed model is consistent with the absence of a periodic repeat unit along the chain direction, with the h k 0 reflections seen in wide-angle X-ray and electron diffraction, with the spherulitic morphology reported in some studies, and with the platelike morphologies obtained under some conditions of precipitation from dilute solution.     

Oxidative stabilization of acrylic fibres

The mechanism of oxidative stabilization of acrylic fibres is characterized by two limiting cases which are determined by the fibre chemistry, the reaction conditions, and the diameter of the filament. These limiting cases correspond to diffusion-limited and reaction-limited kinetic processes. Although the chemistry of stabilization is too complex to specify, the various reactions are separated into two categories: those which occur prior to or concurrently with polymerization of the nitrile groups, called prefatory reactions; and those which occur subsequent to nitrile polymerization, called sequent reactions. Under conditions which allow the prefatory reactions to occur significantly before the sequent reactions, the diffusion of oxygen to reactive sites is limited by previously oxidized material; and the fibre shows a typical two-zone morphology. Under conditions where the prefatory and sequent reactions occur sequentially, the overall stabilization process is limited by the rate of the prefatory reactions; but a skin is established at the fibre surface which acts as an oxygen barrier. Data from a variety of sources, including oxygen analysis, microscopic examination, fibre residue after etching, tension developed in fibres held at constant length, and small-angle X-ray patterns, are cited as evidence for the two limiting cases.Based in part on a thesis submitted by SBW in partial fulfilment of the requirements for the Sc.D. degree in materials engineering, MIT, 1976;     

Oxidative desulfurization: kinetic modelling

Toxicity of hexavalent chromium (Cr(6+)) was focused on with a publication of EU RoHS directive, a novel method to determine hexavalent chromium is developed. It is a combination of energy dispersion X-ray fluorescence spectrometry (EDXRF), spot test, alkali digestion and UV-vis spectrophotometric analysis. First, by EDXRF screening, the presence or absence of element Cr was established. Spot test was followed to identify the valent state of chromium because Cr(6+) and Cr(3+) normally coexist. After alkali digestion, Cr(VI) was separated without an undersired Cr(VI)-Cr(III) interconversions. With a color reagent (DPC) to chelated with Cr(VI), the solution was finally detected by a UV-vis spectrophotometer at a wavelength of 540 nm which is the basis of analyzing Cr(VI) quantitatively. Some parameters affecting analyses were studied. It was found that when pH in the final solution was 2.0, the extraction time was 60 min, the extraction temperature was 90 degrees C, pH during the extraction process was 7.5-8.5, and a mixed buffer solution (0.5M K(2)HPO(4)/0.5M KH(2)PO(4)) was added up to 1 ml, colorimetric reagent was added to 2 ml, it is optimal for extraction. Under this condition, interferences from Fe(3+), Pb(2+), Ag(+), etc., were overcome. It was also found that the curves are rectilinear in the range of 0-500 microg l(-1), the correlation coefficient is up to 0.999924, and the recovery rates are more than 85%, the Cr(III)-DPCO complex can be kept stable for 24h with a relative humidity (RH) range of 60-90%, and a temperature range of 5-40 degrees C. So it can be concluded that the proposed method has a good sensitivity and high precision. It is a more convincing and reliable method due to its relative standard deviation (R.S.D.) <1% after six replicate determinations of Cr(VI) in an Fe-Ni alloy sample.     

Oxidative stabilization of acrylic fibres

When acrylic fibres are heat treated for various times at 220 to 250° C, they form dark, insoluble structures of uncertain chemical character which are inert to many strong oxidizing and reducing agents. The heat-treated fibres are, however, rapidly decoloured by warm alkaline hypochlorite solutions. When fibres which have undergone short-time heat treatment are subjected to the hypochlorite, incubation periods are observed before decolouration is noted; and a swollen acrylic network remains after decolouration is complete. The acrylic network is primarily unreacted precursor units save for a small amount of hydrolyzed material. The decoloured reaction is zero order, indicating a reaction at the surface. The rate of the decolouration reaction also increases with increasing duration of the stabilization heat treatment. In fibres which have undergone partial diffusion-controlled stabilization, a dark mantle surrounds a lightly coloured core. The rate of decolouration is unaffected as the decolouration interface passes from the mantle to the core, indicating that the decolouration reaction is not influenced by the occurance of any sequent reactions. The existence of the acrylic residue indicates that the prefatory reactions are continuing in both mantle and core during the course of stabilization.¹³C-NMR spectra of the acrylic residue show the same triad methine peak areas as those obtained on the untreated fibre; hence the stereoregularity of the nitrile groups has no influence on the rate of nitrile polymerization. The mechanisms of nitrile initiation and of decolouration are discussed. The residue obtained by sulphuric acid etch is different from that obtained by hypochlorite treatment. These results suggest that during the early-to-intermediate stages of stabilization, the fibre consists of interpenetrating networks of original material, i.e., fibre which has undergone only the prefatory reactions and fibre which has undergone the sequent reactions.     

X射线成像术(上)

该文着重从衬度的形成讨论了X射线成像。介绍衬度生成的三种机制:吸收、位相变动和衍射。还介绍了成像设备的主要构成部件:X射线源(X射线发生器、直线加速器和同步辐射),各类探测器、像增强器与显示器,机械、控制与数据处理系统等;应用各种衬度形成的一些成像方法也作了简述,如利用吸收衬度的造影成像、数字减影和双色减影成像、计算机断层成像;利用相位衬度的干涉仪法、类同轴全息法和衍射增强法;利用衍射衬度的Lang透射法和Berg-Barrett反射法等,并用少量例子说明。     

X射线成像术(下)

该文的前半部分(本刊上一期)已扼要介绍了X射线成像的三种衬度机制及成像设备各主要组件的构造,下半部分将继续介绍应用各种衬度的不同的成像方法和一些实例。     

Multimodal-Synergistic-Modulation Neuromorphic Imaging Systems for Simulating Dry Eye Imaging

Inspired by human eyes, the neuromorphic visual system employs a highly efficient imaging and recognition process, which offers tremendous advantages in image acquisition, data pre-processing, and dynamic storage. However, it is still an enormous challenge to simultaneously simulate the structure, function, and environmental adaptive behavior of the human eye based on one device. Here, a multimodal-synergistic-modulation neuromorphic imaging system based on ultraflexible synaptic transistors is successfully presented and firstly simulates the dry eye imaging behavior at the device level. Moreover, important functions of the human visual system in relation to optoelectronic synaptic plasticity, image erasure and enhancement, real-time preprocessing, and dynamic storage are simulated by versatile devices. This work not only simplifies the complexity of traditional neuromorphic visual systems, but also plays a positive role in the publicity of biomedical eye care.