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https://www.readbyqxmd.com/read/28107381/in-vivo-two-photon-imaging-of-astrocytes-in-gfap-gfp-transgenic-mice
#1
Dongjun Guo, Jia Zou, Nicholas Rensing, Michael Wong
Astrocytes play important roles in normal brain function and neurological diseases. In vivo two-photon excitation laser scanning microscopy has the potential to reveal rapid, dynamic structural changes in cells in a variety of physiological and pathological conditions. The type of in vivo imaging method has been shown to affect the plasticity of dendritic spines of neurons, but the optimal in vivo imaging methods of astrocytes have not been established. We compared open-skull and thinned-skull imaging methods for two-photon laser microscopy of live astrocytes in neocortex of GFAP-GFP transgenic mice...
2017: PloS One
https://www.readbyqxmd.com/read/28106206/moo3-x-quantum-dots-for-photoacoustic-imaging-guided-photothermal-photodynamic-cancer-treatment
#2
Dandan Ding, Wei Guo, Chongshen Guo, Jianzhe Sun, Nannan Zheng, Fei Wang, Mei Yan, Shaoqin Liu
A theranostic system of image-guided phototherapy is considered as a potential technique for cancer treatment because of the ability to integrate diagnostics and therapies together, thus enhancing accuracy and visualization during the treatment. In this work, we realized photoacoustic (PA) imaging-guided photothermal (PT)/photodynamic (PD) combined cancer treatment just via a single material, MoO3-x quantum dots (QDs). Due to their strong NIR harvesting ability, MoO3-x QDs can convert incident light into hyperthermia and sensitize the formation of singlet oxygen synchronously as evidenced by in vitro assay, hence, they can behave as both PT and PD agents effectively and act as a "dual-punch" to cancer cells...
January 20, 2017: Nanoscale
https://www.readbyqxmd.com/read/28106175/a-bioorthogonal-turn-on-fluorescent-probe-for-tracking-mitochondrial-nitroxyl-formation
#3
Kyoung Sunwoo, Kondapa Naidu Bobba, Ja-Yun Lim, Taegun Park, Arup Podder, June Seok Heo, Seung Gwan Lee, Sankarprasad Bhuniya, Jong Seung Kim
A bioreductant-resistant 'turn-on' chemodosimetric fluorescent probe Mito-1 has been developed for the detection of mitochondrial HNO in live cells. Mito-1 enables the detection of HNO as low as ∼18 nM. It has the capability to detect both exogenous and endogenous mitochondrial HNO formations in cellular milieus by providing fluorescence images. Its two-photon imaging ability fosters its use as a noninvasive imaging tool for the detection of mitochondrial nitroxyl.
January 20, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28106114/multiplexing-pka-and-erk1-2-kinases-fret-biosensors-in-living-cells-using-single-excitation-wavelength-dual-colour-flim
#4
Claire Demeautis, François Sipieter, Julien Roul, Catherine Chapuis, Sergi Padilla-Parra, Franck B Riquet, Marc Tramier
Monitoring of different signalling enzymes in a single assay using multiplex biosensing provides a multidimensional workspace to elucidate biological processes, signalling pathway crosstalk, and determine precise sequence of events at the single living cell level. In this study, we interrogate the complexity in cAMP/PKA-MAPK/ERK1&2 crosstalk by using multi-parameter biosensing experiments to correlate biochemical activities simultaneously in time and space. Using a single excitation wavelength dual colour FLIM method we are able to detect fluorescence lifetime images of two donors to simultaneously measure PKA and ERK1&2 kinase activities in the same cellular localization by using FRET biosensors...
January 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28105835/silver-nanoclusters-beacon-as-stimuli-responsive-versatile-platform-for-multiplex-dnas-detection-and-aptamer-substrate-complexes-sensing
#5
Guoliang Liu, Jingjing Li, Da-Qian Feng, Jun-Jie Zhu, Wei Wang
An activatable silver nanoclusters beacon (ASNCB) was synthesized through a facile one-pot approach and applied for multiplex DNAs, small molecule, and protein sensing. Multifunctional single-stranded DNA sequences are rationally designed and used for ASNCB in situ synthesis. Via target-responsive structure transformation of ASNCB, target recognition induced ASNCB conformational transition and lit up the fluorescent signal of silver nanoclusters. By further implementing two different color ASNCBs (520 and 600 nm), the parallel multiplexed analysis of two target genes (Influenza A virus genes H1N1 and H5N1) is achieved...
January 3, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28103687/live-cell-imaging-of-endogenous-mrna-using-rna-based-fluorescence-turn-on-probe
#6
Wei Qiang Ong, Y Rose Citron, Sayaka Sekine, Bo Huang
Messenger RNA (mRNA) plays a critical role in cellular growth and development. However, there have been limited methods available to visualize endogenous mRNA in living cells with ease. We have designed RNA-based fluorescence "turn-on" probes that target mRNA by fusing an unstable form of Spinach with target-complementary sequences. These probes have been demonstrated to be selective, stable, and capable of targeting various mRNAs for live E. coli imaging.
January 20, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28103663/biological-photothermal-nanodots-based-on-self-assembly-of-peptide-porphyrin-conjugates-for-antitumor-therapy
#7
Qianli Zou, Manzar Abbas, Luyang Zhao, Shukun Li, Guizhi Shen, Xuehai Yan
Photothermal agents can harvest light energy and convert it into heat, offering a targeted and remote-controlled way to destroy carcinomatous cells and tissues. Inspired by the biological organization of polypeptides and porphyrins in living systems, here we have developed a supramolecular strategy to fabricate photothermal nanodots through peptide-modulated self-assembly of photoactive porphyrins. The self-assembling nature of porphyrins induces the formation of J-aggregates as substructures of the nanodots, and thus enables the fabrication of nanodots with totally inhibited fluorescence emission and singlet oxygen production, leading to a high light-to-heat conversion efficiency of the nanodots...
January 19, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28103302/crude-extracts-flavokawain-b-and-alpinetin-compounds-from-the-rhizome-of-alpinia-mutica-induce-cell-death-via-uck2-enzyme-inhibition-and-in-turn-reduce-18s-rrna-biosynthesis-in-ht-29-cells
#8
Ibrahim Malami, Ahmad Bustamam Abdul, Rasedee Abdullah, Nur Kartinee Bt Kassim, Rozita Rosli, Swee Keong Yeap, Peter Waziri, Imaobong Christopher Etti, Muhammad Bashir Bello
Uridine-cytidine kinase 2 is an enzyme that is overexpressed in abnormal cell growth and its implication is considered a hallmark of cancer. Due to the selective expression of UCK2 in cancer cells, a selective inhibition of this key enzyme necessitates the discovery of its potential inhibitors for cancer chemotherapy. The present study was carried out to demonstrate the potentials of natural phytochemicals from the rhizome of Alpinia mutica to inhibit UCK2 useful for colorectal cancer. Here, we employed the used of in vitro to investigate the effectiveness of natural UCK2 inhibitors to cause HT-29 cell death...
2017: PloS One
https://www.readbyqxmd.com/read/28102310/coordinated-cell-motility-is-regulated-by-a-combination-of-lkb1-farnesylation-and-kinase-activity
#9
S Wilkinson, Y Hou, J T Zoine, J Saltz, C Zhang, Z Chen, L A D Cooper, A I Marcus
Cell motility requires the precise coordination of cell polarization, lamellipodia formation, adhesion, and force generation. LKB1 is a multi-functional serine/threonine kinase that associates with actin at the cellular leading edge of motile cells and suppresses FAK. We sought to understand how LKB1 coordinates these multiple events by systematically dissecting LKB1 protein domain function in combination with live cell imaging and computational approaches. We show that LKB1-actin colocalization is dependent upon LKB1 farnesylation leading to RhoA-ROCK-mediated stress fiber formation, but membrane dynamics is reliant on LKB1 kinase activity...
January 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28100648/multi-scale-structural-analysis-of-plant-er-pm-contact-sites
#10
Heather E McFarlane, Eun Kyoung Lee, Laura S van Bezouwen, Bradford Ross, Abel Rosado, A Lacey Samuels
Membrane contact sites (MCS) are recognized across eukaryotic systems as important nanostructures. Endoplasmic reticulum (ER)-plasma membrane (PM) contact sites (EPCS) are involved in excitation-contraction coupling, signalling, and plant responses to stress. In this report, we perform a multi-scale structural analysis of Arabidopsis EPCS that combine live cell imaging, quantitative transmission electron microscopy (TEM), and electron tomography over a developmental gradient. To place EPCS in the context of the entire cortical ER, we examined GFP-HDEL in living cells over a developmental gradient, then SYT1-GFP was used as a specific marker of EPCS...
January 18, 2017: Plant & Cell Physiology
https://www.readbyqxmd.com/read/28100485/quantification-of-growth-factor-signaling-and-pathway-crosstalk-by-live-cell-imaging
#11
Sean M Gross, Peter Rotwein
Peptide growth factors stimulate cellular responses through activation of their trans-membrane receptors. Multiple intracellular signaling cascades are engaged following growth factor - receptor binding, leading to short- and long-term biological effects. Each receptor-activated signaling pathway does not act in isolation, but rather interacts at different levels with other pathways to shape signaling networks that are distinctive for each growth factor. To gain insights into the specifics of growth factor-regulated interactions among different signaling cascades, we developed a HeLa cell line stably expressing fluorescent live-cell imaging reporters that are readouts for two major growth factor-stimulated pathways, Ras - Raf - Mek - Erk and PI3-kinase - Akt...
January 18, 2017: American Journal of Physiology. Cell Physiology
https://www.readbyqxmd.com/read/28098984/red-emitting-ruthenium-ii-and-iridium-iii-complexes-as-phosphorescent-probes-for-methylglyoxal-in-vitro-and-in-vivo
#12
Wenzhu Zhang, Feiyue Zhang, Yong-Lei Wang, Bo Song, Run Zhang, Jingli Yuan
Transition-metal complexes, ruthenium(II) and iridium(III) complexes in particular, with fascinating triplet emissions are rapidly emerging as important phosphorescent dyes for application in the sensing and imaging of biological makers in live cells and organisms. In this contribution, two red-emitting transition-metal complexes, [Ru(bpy)2(DA-phen)](PF6)2 and [Ir(ppy)2(DA-phen)](PF6) (bpy = 2,2'-bipyridine, DA-phen = 4,5-diamino-1,10-phenanthroline, and ppy = 2-phenylpyridine), were designed and synthesized as phosphorescent probes for the highly sensitive and selective detection of methylglyoxal (MGO), an essential biomarker in the etiopathogenesis of several diseases...
January 18, 2017: Inorganic Chemistry
https://www.readbyqxmd.com/read/28098307/a-ratiometric-fluorescence-probe-for-imaging-sulfur-dioxide-derivatives-in-the-mitochondria-of-living-cells
#13
Yongfei Wang, Qingtao Meng, Run Zhang, Hongmin Jia, Xuehu Zhang, Zhiqiang Zhang
Although sulfur dioxide (SO2) plays an essential role in several physiological processes, monitoring of intracellular SO2 at subcellular levels remains challenging due to the lack of rapid and sensitive methods for its quantification in a 100% aqueous solution. Herein, a new hemicyanine dyes-based fluorescence probe, NBD-Id, was designed and synthesized for the detection of SO2 derivatives in pure aqueous solution and living cells. By virtue of a specific 1,4-addition reaction of SO3(2-)HSO3(-) and the polymethine chain of hemicyanine, significant changes in the absorption and fluorescence emission spectra were observed in less than 20 seconds...
January 18, 2017: Organic & Biomolecular Chemistry
https://www.readbyqxmd.com/read/28098271/a-two-channel-responsive-fluorescent-probe-with-aie-characteristics-and-its-application-for-selective-imaging-of-superoxide-anions-in-living-cells
#14
Xiaoying Gao, Guangxue Feng, Purnima Naresh Manghnani, Fang Hu, Nan Jiang, Jianzhao Liu, Bin Liu, Jing Zhi Sun, Ben Zhong Tang
A two-channel responsive and AIE-active fluorescent probe was developed to selectively detect superoxide anions in living cells, which can be used to track the endogenous superoxide anion level when cells undergo apoptosis and inflammation.
January 18, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28095620/tools-for-live-imaging-of-active-rho-gtpases-in-xenopus
#15
REVIEW
Rachel E Stephenson, Ann L Miller
Rho family GTPases are signaling molecules that orchestrate cytoskeletal dynamics in a variety of cellular processes. Because they effect localized changes to the cytoskeleton only in their active (GTP-bound) conformation, the ability to monitor the active state of Rho GTPases in space and time is critical for understanding their function. Here, we summarize popular tools used for live imaging of active Rho GTPases, outlining advantages and drawbacks of these approaches. Additionally, we highlight key features of the Xenopus laevis embryo that make it well-suited for epithelial cell biology and discuss how application of Rho activity reporters in the Xenopus laevis embryo led to the discovery of a novel phenomenon, junctional Rho flares...
January 17, 2017: Genesis: the Journal of Genetics and Development
https://www.readbyqxmd.com/read/28095612/xenopus-laevis-as-a-model-system-to-study-cytoskeletal-dynamics-during-axon-pathfinding
#16
REVIEW
Paula G Slater, Laurie Hayrapetian, Laura Anne Lowery
The model system, Xenopus laevis, has been used in innumerable research studies and has contributed to the understanding of multiple cytoskeletal components, including actin, microtubules, and neurofilaments, during axon pathfinding. Xenopus developmental stages have been widely characterized, and the Xenopus genome has been sequenced, allowing gene expression modifications through exogenous molecules. Xenopus cell cultures are ideal for long periods of live imaging because they are easily obtained and maintained, and they do not require special culture conditions...
January 17, 2017: Genesis: the Journal of Genetics and Development
https://www.readbyqxmd.com/read/28095175/evaluation-of-the-droplet-microarray-platform-for-high-throughput-screening-of-suspension-cells
#17
Anna A Popova, Claire Depew, Katya Manuella Permana, Alexander Trubitsyn, Ravindra Peravali, Jorge Ángel González Ordiano, Markus Reischl, Pavel A Levkin
Phenotypic cell-based high-throughput screenings play a central role in drug discovery and toxicology. The main tendency in cell screenings is the increase of the throughput and decrease of reaction volume in order to accelerate the experiments, reduce the costs, and enable screenings of rare cells. Conventionally, cell-based assays are performed in microtiter plates, which exist in 96- to 1536-wells formats and cannot be further miniaturized. In addition, performing screenings of suspension cells is associated with risk of losing cell content during the staining procedures and incompatibility with high-content microscopy...
November 1, 2016: Journal of Laboratory Automation
https://www.readbyqxmd.com/read/28092786/imaging-thiol-redox-status-in-murine-tumors-in-vivo-with-rapid-scan-electron-paramagnetic-resonance
#18
Boris Epel, Subramanian V Sundramoorthy, Martyna Krzykawska-Serda, Matthew C Maggio, Mark Tseytlin, Gareth R Eaton, Sandra S Eaton, Gerald M Rosen, Joseph P Y Kao, Howard J Halpern
Thiol redox status is an important physiologic parameter that affects the success or failure of cancer treatment. Rapid scan electron paramagnetic resonance (RS EPR) is a novel technique that has shown higher signal-to-noise ratio than conventional continuous-wave EPR in in vitro studies. Here we used RS EPR to acquire rapid three-dimensional images of the thiol redox status of tumors in living mice. This work presents, for the first time, in vivo RS EPR images of the kinetics of the reaction of (2)H,(15)N-substituted disulfide-linked dinitroxide (PxSSPx) spin probe with intracellular glutathione...
December 31, 2016: Journal of Magnetic Resonance
https://www.readbyqxmd.com/read/28092560/diffeomorphic-multi-frame-non-rigid-registration-of-cell-nuclei-in-2d-and-3d-live-cell-images
#19
Marco Tektonidis, Karl Rohr
To gain a better understanding of cellular and molecular processes it is important to quantitatively analyze the motion of subcellular particles in live cell microscopy image sequences. Since generally the subcellular particles move and cell nuclei move as well as deform, it is important to decouple the movement of particles from that of the cell nuclei using nonrigid registration methods. We have developed a diffeomorphic multi-frame approach for non-rigid registration of cell nuclei in 2D and 3D live cell fluorescence microscopy images...
January 16, 2017: IEEE Transactions on Image Processing: a Publication of the IEEE Signal Processing Society
https://www.readbyqxmd.com/read/28092374/the-movers-and-shapers-in-immune-privilege-of-the-cns
#20
REVIEW
Britta Engelhardt, Peter Vajkoczy, Roy O Weller
Discoveries leading to an improved understanding of immune surveillance of the central nervous system (CNS) have repeatedly provoked dismissal of the existence of immune privilege of the CNS. Recent rediscoveries of lymphatic vessels within the dura mater surrounding the brain, made possible by modern live-cell imaging technologies, have revived this discussion. This review emphasizes the fact that understanding immune privilege of the CNS requires intimate knowledge of its unique anatomy. Endothelial, epithelial and glial brain barriers establish compartments in the CNS that differ strikingly with regard to their accessibility to immune-cell subsets...
February 2017: Nature Immunology
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