photoconversion

光转换
  • 文章类型: Journal Article
    调节自噬和线粒体自噬,至关重要的细胞质量控制系统,为重大疾病提供治疗潜力。然而,有限的药物筛选方案阻碍了进展。我们提出了一种使用光开关荧光报告子的新测定法,mito-kaede,量化线粒体自噬通量。Mito-Kaede优越的紫外线诱导的光转换和亮度转换后,使其成为延长线粒体动力学跟踪的理想选择。它对线粒体自噬反应的特异性,通过parkin敲除细胞证实,增加价值。当与定制的流体交换系统结合时,实现有效的媒介变化,精确的线粒体自噬观察变得可行。这个线粒体自噬试验,除了我们的方法论见解,可以破译线粒体自噬在病理学中的作用,并支持药物筛选工作。
    我们的方法介绍了一种新颖的系统方法,用于按时间顺序跟踪可光活化荧光蛋白的荧光衰减,Mito-Kaede.这与流体交换方法相结合,可以在线粒体自噬刺激之前和之后进行定点观察。
    Modulating autophagy and mitophagy, vital cellular quality control systems, offer therapeutic potential for critical illnesses. However, limited drug screening options hinder progress. We present a novel assay using the photoswitchable fluorescent reporter, mito-Kaede, to quantify mitophagy flux. Mito-Kaede\'s superior UV-induced photoconversion and brightness post-conversion make it ideal for prolonged mitochondrial dynamics tracking. Its specificity in responding to mitophagy, confirmed by parkin-knockout cells, adds value. When coupled with a custom fluid exchange system, enabling efficient medium changes, precise mitophagy observations become feasible. This mitophagy assay, alongside our methodological insights, can decipher mitophagy\'s role in pathology and supports drug screening efforts.
    Our method introduces a novel systematic approach for chronologically tracking the fluorescent decay of a photoactivatable fluorescent protein, mito-Kaede. This is combined with a fluid-exchange method to enable fixed-point observations before and after mitophagy stimulation.
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  • 文章类型: Journal Article
    六个先前未描述的2,9-脱氧类黄酮(1/2a,1/2b,6和7),连同六个已知的化合物(3-5和8-10),是从Aglaiaodorata的树枝和叶子中分离出来的。它们的结构是通过光谱分析的组合确定的,ECD计算和酶水解测定。令人惊讶的是,(±)aglaindanoneE(11a,11b)在暴露于环境自然光时出乎意料地形成为化合物3-6的衍生物。此外,提出了2,9-脱氧黄酮的合理生物合成途径,并进行了化学模拟。在生物活性测定中,化合物1/2a,1/2b,图4和6在0.75%CSE诱导的BEAS-2B细胞损伤模型中显示出潜在的保护作用。
    Six previously undescribed 2, 9-deoxyflavonoids (1/2a, 1/2b, 6, and 7), along with six known compounds (3-5, and 8-10), were isolated from the twigs and leaves of Aglaia odorata. Their structures were determined by a combination of spectral analysis, ECD calculation and enzymatic hydrolysis assay. Surprisingly, (±) aglaindanone E (11a, 11b) were unexpectedly formed as the derivatives of compounds 3-6 when exposed to ambient natural light. Furthermore, the plausible biosynthetic pathway of 2, 9-deoxyflavonoids was proposed and chemically mimicked. In biological activity assay, compounds 1/2a, 1/2b, 4, and 6 showed potential protective effects in the 0.75%CSE-induced BEAS-2B cells injury model.
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  • 文章类型: Journal Article
    铝石榴石在掺入不匹配元素方面表现出卓越的适应性,从而促进具有定制性能的新型材料的合成。这项研究探索了Ce3掺杂的Tb3Al5-xScxO12晶体(其中x范围从0.5到3.0),揭示了一种通过原子尺寸失配工程控制发光和光转换的新方法。拉曼光谱证实了石榴石和钙钛矿相的共存,Sc取代显着影响石榴石晶格,并诱导A1g模式软化至Sc浓度x=2.0。Sc原子控制亚共晶夹杂物的形成,创建有效的光散射中心,并揭示八面体位置饱和的成分阈值。这种调制能够控制Ce3+和Tb3+离子之间的能量转移动力学,增强发光和减轻猝灭。Sc混合过程调节发光效率(LE),显色指数(CRI),和相关色温(CCT),CRI从68调整到84,CCT从3545K调整到12,958K。掺杂Ce3+的Tb3Al5-xScxO12晶体(其中x=2.0)达到了114.6lm/W的最高LE,并以4942K的CCT发射光,类似于白日。这种方法可以设计和开发具有适用于照明技术的定制光学特性的功能材料,持久性磷光体,闪烁体,和储存磷光体。
    Aluminum garnets display exceptional adaptability in incorporating mismatching elements, thereby facilitating the synthesis of novel materials with tailored properties. This study explored Ce3+-doped Tb3Al5-xScxO12 crystals (where x ranges from 0.5 to 3.0), revealing a novel approach to control luminescence and photoconversion through atomic size mismatch engineering. Raman spectroscopy confirmed the coexistence of garnet and perovskite phases, with Sc substitution significantly influencing the garnet lattice and induced A1g mode softening up to Sc concentration x = 2.0. The Sc atoms controlled sub-eutectic inclusion formation, creating efficient light scattering centers and unveiling a compositional threshold for octahedral site saturation. This modulation enabled the control of energy transfer dynamics between Ce3+ and Tb3+ ions, enhancing luminescence and mitigating quenching. The Sc admixing process regulated luminous efficacy (LE), color rendering index (CRI), and correlated color temperature (CCT), with adjustments in CRI from 68 to 84 and CCT from 3545 K to 12,958 K. The Ce3+-doped Tb3Al5-xScxO12 crystal (where x = 2.0) achieved the highest LE of 114.6 lm/W and emitted light at a CCT of 4942 K, similar to daylight white. This approach enables the design and development of functional materials with tailored optical properties applicable to lighting technology, persistent phosphors, scintillators, and storage phosphors.
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  • 文章类型: Journal Article
    追踪整个群体中的单个细胞通路对于理解他们的行为至关重要,细胞通讯,迁移动态,和命运。光学标记是追踪单个细胞的一种方法,但它通常需要遗传修饰来诱导光转化蛋白的产生。然而,这种方法有局限性,不适用于某些细胞类型。例如,遗传修饰常导致巨噬细胞死亡。这项研究旨在通过利用光可转换的微米大小的胶囊来开发一种标记巨噬细胞的替代方法,该胶囊能够容易内化并延长细胞内的滞留时间。在聚乙烯醇凝胶介质中的热处理用于具有多种荧光染料的胶囊的可扩展合成,包括罗丹明6G,维罗宁B,荧光素,吖啶黄,吖啶橙,噻嗪红,以前报道过罗丹明B的荧光亮度,光稳定性,使用共聚焦激光扫描显微镜评估胶囊的光转换能力。活力,摄取,移动性,在RAW264.7和骨髓来源的巨噬细胞上进行光转换研究,作为模型细胞系。由于使用聚乙烯醇凝胶,胶囊的产量增加,消除了对常规过滤步骤的需要。包埋罗丹明B和罗丹明6G的胶囊满足在单个细胞追踪中细胞内使用的所有要求。质谱分析揭示了一系列脱乙基步骤,这些步骤在辐照时导致染料光谱中的蓝移。对巨噬细胞的细胞研究表明胶囊的强烈摄取。胶囊表现出最小的细胞毒性,对细胞运动性的影响可忽略不计。细胞内含有RhB的胶囊的成功光转换凸显了它们作为单个细胞标记的光转换蛋白的替代品的潜力。在个性化医疗中具有广阔的应用前景。
    Tracing individual cell pathways among the whole population is crucial for understanding their behavior, cell communication, migration dynamics, and fate. Optical labeling is one approach for tracing individual cells, but it typically requires genetic modification to induce the generation of photoconvertible proteins. Nevertheless, this approach has limitations and is not applicable to certain cell types. For instance, genetic modification often leads to the death of macrophages. This study aims to develop an alternative method for labeling macrophages by utilizing photoconvertible micron-sized capsules capable of easy internalization and prolonged retention within cells. Thermal treatment in a polyvinyl alcohol gel medium is employed for the scalable synthesis of capsules with a wide range of fluorescent dyes, including rhodamine 6G, pyronin B, fluorescein, acridine yellow, acridine orange, thiazine red, and previously reported rhodamine B. The fluorescence brightness, photostability, and photoconversion ability of the capsules are evaluated using confocal laser scanning microscopy. Viability, uptake, mobility, and photoconversion studies are conducted on RAW 264.7 and bone marrow-derived macrophages, serving as model cell lines. The production yield of the capsules is increased due to the use of polyvinyl alcohol gel, eliminating the need for conventional filtration steps. Capsules entrapping rhodamine B and rhodamine 6G meet all requirements for intracellular use in individual cell tracking. Mass spectrometry analysis reveals a sequence of deethylation steps that result in blue shifts in the dye spectra upon irradiation. Cellular studies on macrophages demonstrate robust uptake of the capsules. The capsules exhibit minimal cytotoxicity and have a negligible impact on cell motility. The successful photoconversion of RhB-containing capsules within cells highlights their potential as alternatives to photoconvertible proteins for individual cell labeling, with promising applications in personalized medicine.
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  • 文章类型: Journal Article
    由于其大的疏水性,可以在悬浮颗粒和沉积物中广泛检测到十氯联苯(PCB-209),和它的一些转化产品可能通过食物链威胁生物。在这里,我们研究了PCB-209在黄河悬浮颗粒上的光化学转化。发现悬浮颗粒具有明显的屏蔽作用,可以在很大程度上抑制PCB-209的光降解。同时,无机离子(如Mg2+和NO3-)和有机物(如腐殖酸,HA)在黄河水中抑制了反应。PCB-209的主要转化产物为低氯化和羟基化多氯联苯(OH-PCBs),还观察到少量的五氯苯酚(PCP)和多氯二苯并呋喃(PCDF)。通过理论计算,提出了通过双•OH攻击碳桥形成PCP的机理和通过离子态OH-PCBs的消除反应形成PCDF的机理。这为持久性有机污染物之间的相互转化提供了一些新的见解。结合VEGA和EPISuite软件,一些中间体如PCDF对生物体的毒性比PCB-209更大。这项研究加深了对PCB-209在阳光下悬浮颗粒上转化行为的理解。
    Decachlorobiphenyl (PCB-209) can be widely detected in suspended particles and sediments due to its large hydrophobicity, and some of its transformation products may potentially threaten organisms through the food chain. Here we investigate the photochemical transformation of PCB-209 on suspended particles from the Yellow River. It was found that the suspended particles had an obvious shielding effect to largely inhibit the photodegradation of PCB-209. Meanwhile, the presence of inorganic ions (e.g. Mg2+ and NO3-) and organic matters (e.g. humic acid, HA) in the Yellow River water inhibited the reaction. The main transformation products of PCB-209 were lower-chlorinated and hydroxylated polychlorinated biphenyls (OH-PCBs), and small amounts of pentachlorophenol (PCP) and polychlorinated dibenzofurans (PCDFs) were also observed. The mechanisms of PCP formation by double •OH attacking carbon bridge and PCDFs formation by elimination reaction of ionic state OH-PCBs were proposed using theoretical calculations, which provided some new insights into the inter-transformations between persistent organic pollutants. In combination with VEGA and EPI Suite software, some intermediates such as PCDFs were more toxic to organisms than PCB-209. This study deepens the understanding of the transformation behavior of PCB-209 on suspended particles under sunlight.
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  • 文章类型: Journal Article
    蓝细菌色素(CBCR)是与植物色素可逆地感应红光和远红光的蓝细菌光感受器。仅需要cGMP磷酸二酯酶/腺苷酸环化酶/FhlA(GAF)结构域用于发色团掺入和适当的光转化。CBCRGAF结构域共价连接线性四吡咯发色团,并在两个光吸收状态之间显示可逆的光转换。在大多数情况下,两种光吸收状态在黑暗条件下是稳定的,但在某些情况下,在热松弛期间,光产物状态经历热松弛回到暗适应状态。在这项研究中,我们检查了工程化的CBCRGAF域,AnPixJg2_BV4。AnPixJg2_BV4共价结合胆绿素IX-α(BV),并在远红色吸收Pfr暗适应状态和橙色吸收Po光产物状态之间显示可逆的光转换。因为BV是哺乳动物细胞的固有发色团,吸收穿透深层组织的远红光,BV结合CBCR分子可用于开发哺乳动物中使用的光遗传学和生物成像工具。为了获得更好的发育平台分子,我们在Phe319位置进行了位点饱和随机诱变。我们成功地获得了具有更高发色团结合效率和更高摩尔消光系数的变体分子。此外,我们观察到热弛豫动力学的广泛变化,最慢和最快的速度相差81倍。具有相对缓慢和快速热弛豫的两种分子对于光遗传学控制将是有利的。
    Cyanobacteriochromes (CBCRs) are cyanobacterial photoreceptors distantly related to the phytochromes sensing red and far-red light reversibly. Only the cGMP phosphodiesterase/Adenylate cyclase/FhlA (GAF) domain is needed for chromophore incorporation and proper photoconversion. The CBCR GAF domains covalently ligate linear tetrapyrrole chromophores and show reversible photoconversion between two light-absorbing states. In most cases, the two light-absorbing states are stable under dark conditions, but in some cases, the photoproduct state undergoes thermal relaxation back to the dark-adapted state during thermal relaxation. In this study, we examined the engineered CBCR GAF domain, AnPixJg2_BV4. AnPixJg2_BV4 covalently binds biliverdin IX-alpha (BV) and shows reversible photoconversion between a far-red-absorbing Pfr dark-adapted state and an orange-absorbing Po photoproduct state. Because the BV is an intrinsic chromophore of mammalian cells and absorbs far-red light penetrating into deep tissues, BV-binding CBCR molecules are useful for the development of optogenetic and bioimaging tools used in mammals. To obtain a better developmental platform molecule, we performed site-saturation random mutagenesis on the Phe319 position. We succeeded in obtaining variant molecules with higher chromophore-binding efficiency and higher molar extinction coefficient. Furthermore, we observed a wide variation in thermal relaxation kinetics, with an 81-fold difference between the slowest and fastest rates. Both molecules with relatively slow and fast thermal relaxation would be advantageous for optogenetic control.
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  • 文章类型: Journal Article
    背景:DHP→CS光转换首次在电子密度重新分布方面进行了分析。提出了以下对CS松弛时C4a-C4b键不恢复的解释:在此过程中,这些原子对之间的库仑排斥能增加了近一倍半,并且它们在LUMO处通过电子键合不足以恢复C4a-C4b键。根据计算,在CS松弛时,连接苯环的连接体发生显著的结构变化。在这种情况下,C4a和C4b原子之间的距离从3.00±增加到3.28±。计算表明,DHP键的C4a-C4b振动具有非常低的强度。因此,热运动不会导致这种键的断裂。
    方法:所有计算均使用Gaussian16软件包在B3LYP/6-311++G(d,p)/IEFPCM理论水平。B3LYP是Gaussian16支持的唯一杂合功能,它确保了DHP的C4a-C4b键的裂解,同时优化了其S1激发态。使用原子电荷的NPA分析,对所研究的构象异构体中电子密度的重新分布进行了定量描述。环己烷用作隐含指定的非极性溶剂。分子轨道的可视化,和电子密度,以及计算的红外光谱的绘图,使用Gaussview6软件包进行。
    BACKGROUND: DHP → CS photoconversion was analyzed in terms of electron density redistribution for the first time. The following explanation for the non-recovery of the C4a-C4b bond upon CS relaxation is proposed: during this process, the Coulomb repulsion energy between these pairs of atoms increases by almost one and a half times, and their bonding by an electron at LUMO is insufficient to recover the C4a-C4b bond. According to calculations, upon CS relaxation, the linker connecting the benzene rings undergoes significant structural changes. In this case, the distance between the C4a and C4b atoms increases from 3.00 Å to 3.28 Å. Calculations showed that the C4a-C4b vibration of the DHP bond has a very low intensity. Therefore, thermal motion does not contribute to the rupture of this bond.
    METHODS: All calculations were performed using the Gaussian16 software package at the B3LYP/6-311 +  + G(d,p)/IEFPCM theory level. B3LYP was the only hybrid functional supported by Gaussian16, which ensured the cleavage of the C4a-C4b bond of DHP while optimizing its S1 excited state. A quantitative description of the redistribution of electron density in the studied conformers was carried out using the analysis of the NPA of atomic charges. Cyclohexane was used as an implicitly specified non-polar solvent. Visualization of molecular orbitals, and electron densities, as well as plotting of calculated IR spectra, were performed using the Gaussview6 software package.
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  • 文章类型: Journal Article
    近红外(NIR)光因其深层组织渗透而非常适合恶性疾病的光学成像和无线光疗,低自发荧光,微弱的组织散射,和非侵入性。稀土纳米粒子(RENP)是很有前途的近红外响应材料,由于其优异的物理和化学性质。镧系元素的4f电子亚壳层,稀土元素的主要群体,具有丰富的能级结构。这促进了广谱光到光的转换以及将光转换为其他形式的能量,如热能和化学能。此外,表面上丰富的可加载和可修改的位点为RENP的功能扩展提供了有利条件。在这次审查中,作者系统地讨论了RENP对近红外光响应的主要过程和机制,并总结了其在光学成像中应用的最新进展,光热疗法,光动力疗法,光免疫疗法,光遗传学,和光响应性药物释放。最后,考虑了在NIR激活下将RENP应用于光学成像和无线光疗的挑战和机遇。
    Near-infrared (NIR) light is well-suited for the optical imaging and wireless phototherapy of malignant diseases because of its deep tissue penetration, low autofluorescence, weak tissue scattering, and non-invasiveness. Rare earth nanoparticles (RENPs) are promising NIR-responsive materials, owing to their excellent physical and chemical properties. The 4f electron subshell of lanthanides, the main group of rare earth elements, has rich energy-level structures. This facilitates broad-spectrum light-to-light conversion and the conversion of light to other forms of energy, such as thermal and chemical energies. In addition, the abundant loadable and modifiable sites on the surface offer favorable conditions for the functional expansion of RENPs. In this review, the authors systematically discuss the main processes and mechanisms underlying the response of RENPs to NIR light and summarize recent advances in their applications in optical imaging, photothermal therapy, photodynamic therapy, photoimmunotherapy, optogenetics, and light-responsive drug release. Finally, the challenges and opportunities for the application of RENPs in optical imaging and wireless phototherapy under NIR activation are considered.
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  • 文章类型: Journal Article
    2015年,我们报告了启动转换,一种将绿色光转换为红色的光转换荧光蛋白的新方法,它成为精密光学成像的强大工具。启动转换使用蓝光和红光到远红光的拦截,而不是传统的紫光或近紫外光照明,这提供了一系列的优势。这里,我们回顾了引物转换的基本原理和应用,重点是它在单细胞标记和谱系追踪中的应用。我们提供了血统追踪技术的历史视角,从而涵盖了荧光的基本原理,光转换荧光蛋白,并最终启动转换。然后,我们提出了引发转换的分子要求,并展示了如何将其用于双色高保真谱系追踪。Further,我们讨论了引物转换成像工具包的潜在未来发展,该工具包可以有益于发展和疾病进展的研究。本文受版权保护。保留所有权利。
    In 2015, we reported primed conversion, a novel way to convert green-to-red photoconvertible fluorescent proteins, which emerges as a powerful tool for precision optical imaging. Primed conversion uses the intercept of blue and red-to-far-red light instead of traditional violet or near-UV light illumination which offers a series of advantages. Here, we review the fundamental principles and applications of primed conversion with a focus on its use in single-cell labelling and lineage tracing. We provide a historical perspective of lineage tracing techniques, thereby covering basic principles of fluorescence, photoconvertible fluorescent proteins, and eventually primed conversion. We then present the molecular requirements for primed conversion to take place and showcase how it can be used for dual-colour high-fidelity lineage tracing. Further, we discuss potential future developments of the primed conversion imaging toolkit that can benefit the study of both development and disease progression.
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  • 文章类型: Journal Article
    使用稳态荧光光谱法研究了溶剂对光敏药物双氯芬酸(DCF)光化学转化率的影响。在一组纯溶剂中DCF的光化学反应获得的光谱信息表明,DCF的光转化反应速率不仅取决于中等极性,而且取决于氢键。使用各种溶剂化变色模型对溶剂效应进行了定性和定量评估,包括多参数线性回归分析(MLRA)。有趣的是,MLRA结果(R=0.99)表明,光转换率随着溶剂极化率(π*)和H键供体能力(α)的增加而增加,而速率随着氢键受体能力(β)的增加而降低。然而,观察到溶剂酸度与碱度和极化率相比的主要影响。提出并讨论了合理化H键和中等极性对DCF光转换反应影响的假设。
    The solvent effects on the photochemical conversion rate of the photosensitizing drug diclofenac (DCF) were investigated using steady-state fluorescence spectroscopy. The spectral information obtained for the photochemical reaction of DCF in a set of neat solvents demonstrates that the photoconversion reaction rate of DCF is not only medium polarity dependent but also hydrogen-bonding dependent. The solvent effects were qualitatively and quantitatively assessed employing various solvatochromic models, including multi-parameter linear regression analysis (MLRA). Interestingly, the MLRA results (R = 0.99) revealed that the photoconversion rate increases with increasing solvent polarizability (π*) and H-bond donor capability (α), whereas the rate decreases with increasing hydrogen-bond acceptor capability (β). However, predominant effect of the solvent acidity compared to basicity and polarizability was observed. A hypothesis rationalizing the effects of H-bonding and medium polarity on DCF photoconversion reaction is presented and discussed.
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