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https://www.readbyqxmd.com/read/28931067/in-vivo-imaging-system-for-explants-analysis-a-new-approach-for-assessment-of-cell-transplantation-effects-in-large-animal-models
#1
Weronika Zarychta-Wiśniewska, Anna Burdzinska, Radosław Zagozdzon, Bartosz Dybowski, Marta Butrym, Zdzisław Gajewski, Leszek Paczek
INTRODUCTION: Despite spectacular progress in cellular transplantology, there are still many concerns about the fate of transplanted cells. More preclinical studies are needed, especially on large animal models, to bridge the translational gap between basic research and the clinic. Herein, we propose a novel approach in analysis of cell transplantation effects in large animals explants using in vivo imaging system (IVIS®) or similar equipment. MATERIAL AND METHODS: In the in vitro experiment cells labeled with fluorescent membrane dyes: DID (far red) or PKH26 (orange) were visualized with IVIS®...
2017: PloS One
https://www.readbyqxmd.com/read/28930309/a-global-method-for-handling-fluorescence-spectra-at-high-concentration-derived-from-the-competition-between-emission-and-absorption-of-colloidal-cdte-quantum-dots
#2
Thomas Noblet, Laurent Dreesen, Julie Hottechamps, Christophe Humbert
We investigate the effects of the concentration of CdTe quantum dots (QDs) on their fluorescence in water. The emission spectra, acquired in right angle geometry, exhibit highly variable shapes. The measurements evidence a critical value of the concentration beyond which the intensity and the spectral bandwidth decrease and the fluorescence maximum is redshifted. To account for these observations, we develop a model based on the primary and secondary inner filter effects. The accuracy of the model ensures that the concentration dependent behaviour of QD fluorescence is completely due to inner filter effects...
September 20, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28929742/beta-hemolytic-bacteria-selectively-trigger-liposome-lysis-enabling-rapid-and-accurate-pathogen-detection
#3
Rongji Sum, Muthukaruppan Swaminathan, Sahil Kumar Rastogi, Obdulio Piloto, Ian Cheong
For more than a century, blood agar plates have been the only test for beta-hemolysis. Although blood agar cultures are highly predictive for bacterial pathogens, they are too slow to yield actionable information. Here, we show that beta-hemolytic pathogens are able to lyse and release fluorophores encapsulated in sterically stabilized liposomes whereas alpha and gamma-hemolytic bacteria have no effect. By analyzing fluorescence kinetics, beta-hemolytic colonies cultured on agar could be distinguished in real time with 100% accuracy within 6 h...
September 20, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28929568/a-selective-and-sensitive-fluorescent-sensor-for-cysteine-detection-based-on-bi-8-carboxamidoquinoline-derivative-and-cu-2-complex
#4
Guoping Chai, Qingwen Liu, Qiang Fei, Jinling Zhang, Xiaoxiao Sun, Hongyan Shan, Guodong Feng, Yanfu Huan
In this paper, a novel fluorescent sensor 1 for selective and sensitive detection of cysteine was developed based on a complex between bi-8-carboxamidoquinoline derivative ligand (L) and Cu(2+) . The interaction of Cu(2+) with the ligand causes a dramatic fluorescence quenching most likely due to its high affinity towards Cu(2+) and a ligand-metal charge transfer (LMCT) process. The in situ generated L-Cu(2) complex was utilized as a chemosensing ensemble for cysteine. In the presence of cysteine, the fluorophore, L, was released from L-Cu(2) complex because of the strong affinity of cysteine to Cu(2+) via the Cu-S bond, leading to the fluorescence recovery of the ligand...
September 20, 2017: Luminescence: the Journal of Biological and Chemical Luminescence
https://www.readbyqxmd.com/read/28927835/evaluation-of-rice-tetraticopeptide-domain-containing-thioredoxin-as-a-novel-solubility-enhancing-fusion-tag-in-escherichia-coli
#5
Wenjun Xiao, Li Jiang, Weiyu Wang, Ruyue Wang, Jun Fan
Fusion of solubility-enhancing tag is frequently used for improving soluble production of target protein in Escherichia coli. The Arabidopsis tetraticopeptide domain-containing thioredoxin (TDX) has been documented to exhibit functions of disulfide reductase, foldase chaperone, and holdase chaperone. Here, we identified that fusion of rice TDX with the smaller size increased soluble expression levels of three fluorescent proteins with different fluorophores in the E. coli strain BL21(DE3) or the Rosetta (DE3) strain with coexpression of six rare tRNAs, but decreased conformational quality of certain fluorescent proteins, as comparison with the His6-tagged ones...
September 15, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28927677/std-nmr-experiments-identify-a-structural-motif-with-novel-second-site-activity-against-west-nile-virus-ns2b-ns3-protease
#6
Tobias Schöne, Lena Lisbeth Grimm, Naoki Sakai, Linlin Zhang, Rolf Hilgenfeld, Thomas Peters
West Nile virus (WNV) belongs to the genus Flavivirus of the family Flaviviridae. This mosquito-borne virus that is highly pathogenic to humans has been evolving into a global threat during the past two decades. Despite many efforts, neither antiviral drugs nor vaccines are available. The viral protease NS2B-NS3(pro) is essential for viral replication, and therefore it is considered a prime drug target. However, success in the development of specific NS2B-NS3(pro) inhibitors had been moderate so far. In the search for new structural motifs with binding affinity for NS2B-NS3(pro), we have screened a fragment library, the Maybridge Ro5 library, employing saturation transfer difference (STD) NMR experiments as readout...
September 16, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28926050/temperature-dependence-of-the-conversion-efficiency-of-photochromic-perylene-bisimide-dithienylcyclopentene-triads-embedded-in-a-polymer
#7
Johann Thurn, Johannes Maier, Martti Pärs, Katja Gräf, Mukundan Thelakkat, Jürgen Köhler
Photochromic molecules that are covalently linked to a strong fluorophore combine the requirements of external control and strong fluorescence, which will become increasingly important for super-resolution microscopy techniques based on single molecules. However, given the bulky structure of such constructs, steric hindrance might affect their photoconversion efficiencies upon immobilising them for imaging purposes. In this study the efficiencies of the photochromic conversion processes of molecular triads that are embedded in a polymer have been studied as a function of temperature...
September 19, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28925686/a-naphthalimide-based-fluorophore-for-soft-anionic-interface-monitoring
#8
Li Wang, Gang Wang, Congdi Shang, Rui Kang, Yu Fang
A naphthalimide-based low-molecular mass fluorophore (NA) was designed and synthesized by introducing an azetidine unit onto the aromatic ring of the compound as an electron-donating structure, and a hydrophilic (2-(2-aminoethylamino)ethanol) moiety into the 'N-imide site' of the core structure. UV-vis absorption and fluorescence measurements revealed that the fluorophore is soluble in water, and shows a fluorescent quantum yield of ~20% and lifetime of ~3.7 ns in the solvent within a wide pH range. Moreover, the fluorescence emission and anisotropy of the fluorophore as produced are both dependent upon the viscosity and polarity of the medium...
September 19, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28925371/inclusion-of-fluorophores-in-cyclodextrins-a-closer-look-at-the-fluorometric-determination-of-association-constants-by-linear-and-non-linear-fitting-procedures
#9
Rudi Hutterer
The author discusses methods for the fluorometric determination of affinity constants by linear and non-linear fitting methods. This is outlined in particular for the interaction between cyclodextrins and several anesthetic drugs including benzocaine. Special emphasis is given to the limitations of certain fits, and the impact of such studies on enzyme-substrate interactions are demonstrated. Both the experimental part and methods of analysis are well suited for students in an advanced lab.
September 19, 2017: Methods and Applications in Fluorescence
https://www.readbyqxmd.com/read/28925107/formulation-of-long-wavelength-indocyanine-green-nanocarriers
#10
Vikram J Pansare, William J Faenza, Hoang Lu, Douglas H Adamson, Robert K Prud'homme
Indocyanine green (ICG), a Food and Drug Administration (FDA)-approved fluorophore with excitation and emission wavelengths inside the "optical imaging window," has been incorporated into nanocarriers (NCs) to achieve enhanced circulation time, targeting, and real-time tracking in vivo. While previous studies transferred ICG exogenously into NCs, here, a one-step rapid precipitation process [flash nanoprecipitation (FNP)] creates ICG-loaded NCs with tunable, narrow size distributions from 30 to 180 nm. A hydrophobic ion pair of ICG-tetraoctylammonium or tetradodecylammonium chloride is formed either in situ during FNP or preformed then introduced into the FNP feed stream...
September 2017: Journal of Biomedical Optics
https://www.readbyqxmd.com/read/28924668/synthesis-of-janelia-fluor-halotag-and-snap-tag-ligands-and-their-use-in-cellular-imaging-experiments
#11
Jonathan B Grimm, Timothy A Brown, Brian P English, Timothée Lionnet, Luke D Lavis
The development of genetically encoded self-labeling protein tags such as the HaloTag and SNAP-tag has expanded the utility of chemical dyes in microscopy. Intracellular labeling using these systems requires small, cell-permeable dyes with high brightness and photostability. We recently discovered a general method to improve the properties of classic fluorophores by replacing N,N-dimethylamino groups with four-membered azetidine rings to create the "Janelia Fluor" dyes. Here, we describe the synthesis of the HaloTag and SNAP-tag ligands of Janelia Fluor 549 and Janelia Fluor 646 as well as standard labeling protocols for use in ensemble and single-molecule cellular imaging...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28924666/brain-slice-staining-and-preparation-for-three-dimensional-super-resolution-microscopy
#12
Christopher L German, Manasa V Gudheti, Annette E Fleckenstein, Erik M Jorgensen
Localization microscopy techniques-such as photoactivation localization microscopy (PALM), fluorescent PALM (FPALM), ground state depletion (GSD), and stochastic optical reconstruction microscopy (STORM)-provide the highest precision for single-molecule localization currently available. However, localization microscopy has been largely limited to cell cultures due to the difficulties that arise in imaging thicker tissue sections. Sample fixation and antibody staining, background fluorescence, fluorophore photoinstability, light scattering in thick sections, and sample movement create significant challenges for imaging intact tissue...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28924663/quantitative-single-molecule-localization-microscopy-qsmlm-of-membrane-proteins-based-on-kinetic-analysis-of-fluorophore-blinking-cycles
#13
Franziska Fricke, Joel Beaudouin, Sebastian Malkusch, Roland Eils, Mike Heilemann
Photoswitchable or photoactivatable fluorophores are the key in single-molecule localization microscopy. Next to providing fluorescence images with subdiffraction spatial resolution, additional information is available from observing single fluorophores over time. This includes the characteristic photophysical phenomenon of "blinking" that is exhibited by single fluorescent proteins or fluorophores and follows well-defined kinetic laws. Analyzing the kinetics of "blinking" allows determining the number of fluorophores in a multi-molecular complex...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28924659/sted-imaging-of-golgi-dynamics-with-cer-sir-a-two-component-photostable-high-density-lipid-probe-for-live-cells
#14
Roman S Erdmann, Derek Toomre, Alanna Schepartz
Long time-lapse super-resolution imaging in live cells requires a labeling strategy that combines a bright, photostable fluorophore with a high-density localization probe. Lipids are ideal high-density localization probes, as they are >100 times more abundant than most membrane-bound proteins and simultaneously demark the boundaries of cellular organelles. Here, we describe Cer-SiR, a two-component, high-density lipid probe that is exceptionally photostable. Cer-SiR is generated in cells via a bioorthogonal reaction of two components: a ceramide lipid tagged with trans-cyclooctene (Cer-TCO) and a reactive, photostable Si-rhodamine dye (SiR-Tz)...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28924656/live-cell-sted-imaging-with-the-hyper2-biosensor
#15
Natalia M Mishina, Vsevolod V Belousov
Stimulated emission depletion (STED) microscopy is a popular super resolution imaging technique. Not only synthetic dyes and fluorescent proteins can be utilized as STED fluorophores, but also genetically encoded biosensors. Fusing the biosensor with proteins of interest allows subdiffraction imaging of intracellular macromolecular architecture with simultaneous extraction of functional information about cellular activities. Here, we describe a protocol for live-cell STED microscopy of the HyPer2 biosensor fused to cytoskeletal filaments...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28921977/photoinduced-bimolecular-electron-transfer-in-ionic-liquids
#16
Boning Wu, Mark Maroncelli, Edward W Castner
The present work seeks to better understand the role of solute diffusion and solvation dynamics on bimolecular electron transfer in ionic liquids (ILs). Steady-state and time-resolved measurements of the reductive fluorescence quenching of five fluorophores ("F") by six quenchers ("Q"; electron donors) are reported in acetonitrile and two ionic liquids, 1-ethyl-3-methylimidazolium bis(trifluoromethylsulfonyl)amide and trihexyl(tetradecyl)phosphonium bis(trifluoromethylsulfonyl)amide. Data were collected on 66 different F-Q-solvent systems, which span a 2...
September 18, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28920994/a-light-up-endoplasmic-reticulum-probe-based-on-a-rational-design-of-red-emissive-fluorogens-with-aggregation-induced-emission
#17
Chong-Jing Zhang, Xiaolei Cai, Shidang Xu, Ruoyu Zhan, Wu Jien, Bin Liu
Fine-tuning electron acceptors through changing one cyano group to an amide generates a more stable and emissive fluorophore with the character of aggregation-induced emission. Conjugation between the new fluorophore and CFFKDEL generated an excellent ER targeting light-up probe with high specificity and good photostability.
September 18, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28920988/fluorogenic-nir-probes-based-on-1-2-4-5-tetrazine-substituted-bf2-azadipyrromethenes
#18
Dan Wu, Donal F O'Shea
A series of 1,2,4,5-tetrazine integrated near infrared (NIR) fluorophores based on the BF2 azadipyrromethene (NIR-AZA) class has been synthesised and their ability to modulate emission from low to high in response to Diels-Alder cycloaditions has been assessed. Substituents on the tetrazine component of the probe (Cl, OMe, p-NO2C6H4O) were seen to strongly influence quantum yields, fluorescence enhancement factors, and rates of cycloadditions. Cycloadditions between tetrazine-NIR-AZA constructs and a strained alkyne substrate were seen to be highly efficient in organic or aqueous solutions and in gels with high fluorescence enhancements of up to 48-fold observed...
September 18, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28919344/fluorophore-labeling-of-a-cell-penetrating-peptide-induces-differential-effects-on-its-cellular-distribution-and-affects-cell-viability
#19
Ditlev Birch, Malene Vinther Christensen, Dan Staerk, Henrik Franzyk, Hanne Mørck Nielsen
Cell-penetrating peptides constitute efficient delivery vectors, and studies of their uptake and mechanism of translocation typically involve fluorophore-labeled conjugates. In the present study, the influence of a number of specific fluorophores on the physico-chemical properties and uptake-related characteristics of penetratin were studied. An array of seven fluorophores belonging to distinct structural classes was examined, and the impact of fluorophore labeling on intracellular distribution and cytotoxicity was correlated to the physico-chemical properties of the conjugates...
September 14, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28918077/membrane-electro-oxidizer-a-new-hybrid-membrane-system-with-electrochemical-oxidation-for-enhanced-organics-and-fouling-control
#20
Naresh Mameda, Hyung-June Park, Kwang-Ho Choo
The synergistic combination of membrane filtration with advanced oxidation is of particular interest for next-generation wastewater treatment technologies. A membrane electro-oxidizer (MEO) hybridizing a submerged microfilter and an electrochemical cell was developed and investigated for tertiary treatment of secondary industrial (textile) wastewater effluent. Laboratory- and pilot-scale MEO systems were designed and evaluated for treatment efficiency and membrane fouling control. The MEO achieved substantial removal of color (50-90%), turbidity (>90%), and bacteria (>4 log) as well as chemical oxygen demand (13-31%) and 1,4-dioxane (∼25-53%)...
September 5, 2017: Water Research
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