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https://www.readbyqxmd.com/read/28651420/-effects-of-hypoxia-inducible-factor-1%C3%AE-on-p311-and-its-influence-on-the-migration-of-murine-epidermal-stem-cells
#1
Z D Xu, H S Li, S Wang, W F He, J Wu, G X Luo
Objective: To explore the effects of hypoxia inducible factor-1α (HIF-1α) on P311 and its influence on the migration of murine epidermal stem cells (ESCs) under hypoxia in vitro. Methods: Two kinds of murine ESCs were isolated and obtained from 15 neonatal wild-type C57BL/6J mice and 5 congeneric source P311 gene knock-out mice, respectively. The first passage of cells were used in the following experiments after morphologic observation and detection of expression of cell surface markers CD71 and CD49f with flow cytometer...
May 20, 2017: Zhonghua Shao Shang za Zhi, Zhonghua Shaoshang Zazhi, Chinese Journal of Burns
https://www.readbyqxmd.com/read/28651174/label-free-chronopotentiometric-glycoprofiling-of-prostate-specific-antigen-using-sialic-acid-recognizing-lectins
#2
Stefan Belicky, Hana Černocká, Tomas Bertok, Alena Holazova, Kamila Réblová, Emil Paleček, Jan Tkac, Veronika Ostatná
In recent decades, it has become clear that most of human proteins are glycosylated and that protein glycosylation plays an important role in health and diseases. At present, simple, fast and inexpensive methods are sought for clinical applications and particularly for improved diagnostics of various diseases, including cancer. We propose a label- and reagent-free electrochemical method based on chronopotentiometric stripping (CPS) analysis and a hanging mercury drop electrode for the detection of interaction of sialylated protein biomarker a prostate specific antigen (PSA) with two important lectins: Sambucus nigra agglutinin (SNA) and Maackia amurensis agglutinin (MAA)...
June 19, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/28650983/differential-induction-of-apoptosis-and-autophagy-by-pyrrolizidine-alkaloid-clivorine-in-human-hepatoma-huh-7-5-cells-and-its-toxic-implication
#3
Wenju Liu, Xu Li, Bo Zhou, Shoucai Fang, Wenzhe Ho, Hui Chen, Hao Liang, Li Ye, Jun Tang
Growing evidence suggests that the pyrrolizidine alkaloids (PAs)-induced hepatotoxicity is mediated by multiple cell death/defence modalities. However, the detailed mechanisms are still lacking. In this study, the hepatotoxic effects of four PAs including three retronecine-type ones (senecionine, seneciphylline and monocrotaline) and one otonecine-type (clivorine) on the proliferation of Huh-7.5 cells and the possible mechanisms were investigated. The results showed that all the PAs could inhibit cell proliferation and induce apoptosis in a concentration-dependent manner...
2017: PloS One
https://www.readbyqxmd.com/read/28650636/orthogonal-electric-field-measurements-near-the-green-fluorescent-protein-fluorophore-through-stark-effect-spectroscopy-and-pka-shifts-provide-a-unique-benchmark-for-electrostatics-models
#4
Joshua D Slocum, Jeremy T First, Lauren J Webb
Measurement of the magnitude, direction, and functional importance of electric fields in biomolecules has been a longstanding experimental challenge. pKa shifts of titratable residues have been the most widely implemented measurements of local electrostatic environment around the labile proton, and experimental data sets of pKa shifts in a variety of systems have been used to test and refine computational prediction capabilities of protein electrostatic fields. A more direct and increasingly popular technique to measure electric fields in proteins is Stark effect spectroscopy, where the change in absorption energy of a chromophore relative to a reference state is related to the change in electric field felt by the chromophore...
June 26, 2017: Journal of Physical Chemistry. B
https://www.readbyqxmd.com/read/28650487/engineering-thiophene-based-nanoparticles-to-induce-phototransduction-in-live-cells-under-illumination
#5
M Zangoli, F Di Maria, E Zucchetti, C Bossio, M R Antognazza, G Lanzani, R Mazzaro, F Corticelli, M Baroncini, G Barbarella
We report that nanoparticles prepared from appropriately functionalized polythiophenes once administered to live cells can acquire phototransduction properties under illumination, becoming photoactive sites able to absorb visible light and convert it to an electrical signal through cell membrane polarization. Amine-reactive fluorescent nanoparticles with pendant N-succinimidyl-ester groups (NPs-NHS) are prepared from polythiophenes alternating unsubstituted and 3-(2,5-dioxopyrrolidin-1-yl-8-octanoate)-substituted thiophenes by a nanoprecipitation method...
June 26, 2017: Nanoscale
https://www.readbyqxmd.com/read/28650478/a-dna-origami-platform-for-quantifying-protein-copy-number-in-super-resolution
#6
Francesca Cella Zanacchi, Carlo Manzo, Angel S Alvarez, Nathan D Derr, Maria F Garcia-Parajo, Melike Lakadamyali
Single-molecule-based super-resolution microscopy offers researchers a unique opportunity to quantify protein copy number with nanoscale resolution. However, while fluorescent proteins have been characterized for quantitative imaging using calibration standards, similar calibration tools for immunofluorescence with small organic fluorophores are lacking. Here we show that DNA origami, in combination with GFP antibodies, is a versatile platform for calibrating fluorophore and antibody labeling efficiency to quantify protein copy number in cellular contexts using super-resolution microscopy...
June 26, 2017: Nature Methods
https://www.readbyqxmd.com/read/28650461/a-light-and-calcium-gated-transcription-factor-for-imaging-and-manipulating-activated-neurons
#7
Wenjing Wang, Craig P Wildes, Tanyaporn Pattarabanjird, Mateo I Sanchez, Gordon F Glober, Gillian A Matthews, Kay M Tye, Alice Y Ting
Activity remodels neurons, altering their molecular, structural, and electrical characteristics. To enable the selective characterization and manipulation of these neurons, we present FLARE, an engineered transcription factor that drives expression of fluorescent proteins, opsins, and other genetically encoded tools only in the subset of neurons that experienced activity during a user-defined time window. FLARE senses the coincidence of elevated cytosolic calcium and externally applied blue light, which together produce translocation of a membrane-anchored transcription factor to the nucleus to drive expression of any transgene...
June 26, 2017: Nature Biotechnology
https://www.readbyqxmd.com/read/28650154/interrogating-detergent-desolvation-of-nanopore-forming-proteins-by-fluorescence-polarization-spectroscopy
#8
Aaron J Wolfe, Yi-Ching Hsueh, Adam R Blanden, Mohammad M Mohammad, Bach Pham, Avinash Kumar Thakur, Stewart N Loh, Min Chen, Liviu Movileanu
Understanding how membrane proteins interact with detergents is of fundamental and practical significance in structural and chemical biology as well as in nanobiotechnology. Current methods for inspecting protein-detergent complex (PDC) interfaces require high concentrations of protein and are of low throughput. Here, we describe a scalable, spectroscopic approach that uses nanomolar protein concentrations in native solutions. This approach, which is based on steady-state fluorescence polarization (FP) spectroscopy, kinetically resolves the dissociation of detergents from membrane proteins and protein unfolding...
June 26, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28650069/transcriptional-engineering-of-the-glyceraldehyde-3-phosphate-dehydrogenase-promoter-for-improved-heterologous-protein-production-in-pichia-pastoris
#9
Özge Ata, Roland Prielhofer, Brigitte Gasser, Diethard Mattanovich, Pınar Çalık
The constitutive glyceraldehyde-3-phosphate dehydrogenase promoter (PGAP ), which is one of the benchmark promoters of Pichia pastoris, was analyzed in terms of putative transcription factor binding sites. We constructed a synthetic library with distinct regulatory properties through deletion and duplication of these putative transcription factor binding sites and selected transcription factor (TF) genes were overexpressed or deleted to understand their roles on heterologous protein production. Using enhanced green fluorescent protein, an expression strength in a range between 0...
June 26, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28650022/interaction-and-inhibitory-influence-of-the-azo-dye-carmoisine-on-lysozyme-amyloid-fibrillogenesis
#10
Anirban Basu, Gopinatha Suresh Kumar
The binding of the common food colorant carmoisine and its inhibitory effect on amyloid fibrillation in lysozyme have been investigated. Since humans are increasingly exposed to various food colorants like carmoisine, such studies are highly relevant. In the presence of lysozyme, the carmoisine absorption spectrum exhibited hypochromic changes. The intrinsic fluorescence of lysozyme was also quenched on interaction. Time-resolved fluorescence results suggested that the binding mechanism involved ground state complexation...
June 26, 2017: Molecular BioSystems
https://www.readbyqxmd.com/read/28649965/pawflim-reducing-bias-and-uncertainty-to-enable-lower-photon-count-in-flim-experiments
#11
Mauro Silberberg, Hernán E Grecco
Förster resonant energy transfer measured by fluorescence lifetime imaging microscopy (FRET-FLIM) is the method of choice for monitoring the spatio-temporal dynamics of protein interactions in living cells. To obtain an accurate estimate of the molecular fraction of interacting proteins requires a large number of photons, which usually precludes the observation of a fast process, particularly with time correlated single photon counting (TCSPC) based FLIM. In this work, we propose a novel method named pawFLIM (phasor analysis via wavelets) that allows the denoising of FLIM datasets by adaptively and selectively adjusting the desired compromise between spatial and molecular resolution...
June 26, 2017: Methods and Applications in Fluorescence
https://www.readbyqxmd.com/read/28649842/lipo-oligomer-nanoformulations-for-targeted-intracellular-protein-delivery
#12
Peng Zhang, Benjamin Steinborn, Ulrich Lächelt, Stefan Zahler, Ernst Wagner
Here, we report novel lipo-oligoaminoamide nanoformulations for targeted intracellular protein delivery. Formulations are generated by first bioreversibly conjugating a sequence-defined amphiphilic lipo-oligomer 728 to the cargo protein via disulfide bonds, followed by formulation of the formed 728-SS-protein conjugate with different helper lipids in various compositions. The triblock oligoaminoamide 728 contains cysteines for reversible covalent protein conjugation and crosslink-stabilization of formed nanoparticles, polyethylene glycol (PEG) for shielding and providing a hydrophilic domain, eight cationizable succinoyl tetraethylene pentamine (Stp) repeats for endosomal buffering and escape into the cytosol, and a tetra-oleic acid block for hydrophobic stabilization...
June 26, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28649828/dimerization-of-the-conserved-n-terminal-domain-of-a-spider-silk-protein-controls-the-self-assembly-of-the-repetitive-core-domain
#13
Joschka Bauer, Thomas Scheibel
Spider silk proteins comprise a repetitive core domain with polyalanine and glycine/proline stretches flanked by highly conserved nonrepetitive N- and C-terminal domains. The termini are responsive to assembly triggers, sensing changes in the ionic (H+, phosphate) and mechanical (shear stress) environment along the spinning duct. Upon assembly, the N-terminal domain dimerizes in a pH-dependent manner induced by protonation of conserved acidic residues. So far, dimerization of N-terminal spider silk domains has been individually investigated in the absence of large core domains...
June 26, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28649656/lgr5-is-a-marker-for-fetal-mammary-stem-cells-but-is-not-essential-for-stem-cell-activity-or-tumorigenesis
#14
Christy L Trejo, Gidsela Luna, Christopher Dravis, Benjamin T Spike, Geoffrey M Wahl
The search for the bipotent mammary stem cells that drive mammary development requires markers to enable their prospective isolation. There is general agreement that bipotent mouse mammary stem cells are abundant in late fetal development, but their existence in the adult is vigorously debated. Among markers useful for mammary stem cell identification, the Wnt co-receptor Lgr5 has been suggested by some to be both "necessary and sufficient" for bipotency and transplantation of adult mammary stem cell activity, though other studies disagree...
2017: NPJ Breast Cancer
https://www.readbyqxmd.com/read/28649260/tomato-udp-glucose-sterol-glycosyltransferases-a-family-of-developmental-and-stress-regulated-genes-that-encode-cytosolic-and-membrane-associated-forms-of-the-enzyme
#15
Karla Ramirez-Estrada, Nídia Castillo, Juan A Lara, Monserrat Arró, Albert Boronat, Albert Ferrer, Teresa Altabella
Sterol glycosyltransferases (SGTs) catalyze the glycosylation of the free hydroxyl group at C-3 position of sterols to produce sterol glycosides. Glycosylated sterols and free sterols are primarily located in cell membranes where in combination with other membrane-bound lipids play a key role in modulating their properties and functioning. In contrast to most plant species, those of the genus Solanum contain very high levels of glycosylated sterols, which in the case of tomato may account for more than 85% of the total sterol content...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28649045/roles-of-phytoplankton-and-macrophyte-derived-dissolved-organic-matter-in-sulfamethazine-adsorption-on-goethite
#16
Leilei Bai, Chicheng Cao, Chunliu Wang, Changhui Wang, Hui Zhang, Helong Jiang
Phytoplankton-derived dissolved organic matter (PDOM) and macrophyte-derived dissolved organic matter (MDOM) exist ubiquitously in eutrophic freshwater lakes. To understand the heterogeneous roles of individual fluorescent DOM components in the adsorption of antibiotics onto sediment minerals, the adsorptive fractionation of DOM on goethite (α-FeOOH) and its interaction with sulfamethazine (SMT) were investigated using fluorescence excitation-emission matrix combined with parallel factor analysis (EEM-PARAFAC)...
June 22, 2017: Environmental Pollution
https://www.readbyqxmd.com/read/28649039/pm2-5-exposure-in-utero-contributes-to-neonatal-cardiac-dysfunction-in-mice
#17
Vineeta Tanwar, Jeremy M Adelstein, Jacob A Grimmer, Dane J Youtz, Benjamin P Sugar, Loren E Wold
OBJECTIVE: Exposure of fine particulate matter (PM2.5) to pregnant dams has been shown to be strongly associated with adverse cardiovascular outcomes in offspring at adulthood, however, effects evident during neonatal periods are unclear. We designed this study to examine cardiac function of neonatal mice (14 days old) exposed to in utero PM2.5. METHODS: Pregnant FVB female mice were exposed either to filtered air (FA) or PM2.5 at an average concentration of 91...
June 22, 2017: Environmental Pollution
https://www.readbyqxmd.com/read/28649025/a%C3%AE-plaque-selective-nir-fluorescence-probe-to-differentiate-alzheimer-s-disease-from-tauopathies
#18
K Rajasekhar, Nagarjun Narayanaswamy, N Arul Murugan, Keith Viccaro, Hyoung-Gon Lee, Kavita Shah, Thimmaiah Govindaraju
Selective detection and staining of toxic amyloid plaques, a potential biomarker present in the Alzheimer's disease (AD) brain is crucial for both clinical diagnosis and monitoring AD disease progression. Herein, we report a coumarin-quinoline (CQ) conjugate-based turn-on near-infrared (NIR) fluorescence probe for specific detection of β-amyloid (Aβ) aggregates. CQ probe is highly sensitive and exhibits ~100-fold fluorescence enhancement in vitro upon binding Aβ aggregates with enhanced quantum yield. Furthermore, the probe has ~10-fold higher binding affinity towards Aβ aggregates (86nM) compared to commonly used Thioflavin T...
June 17, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28649017/chiral-alkylated-aniline-as-a-noninvasive-fluorescence-sensor-spectroscopic-and-molecular-modeling-studies
#19
Bidisha Sengupta, Chirantan Sen Mukherjee, Sandipan Chakraborty, Maria Jones Muhammad, William Gladney, George Armstrong
Aniline, heterocyclic aromatic amines, and arylamines are known carcinogens. Recently aniline mustard has come into prominence as a novel anticancer agent. In this project, microwave irradiation has been used to synthesize an optically active alkylated aniline namely 2,6-dimethyl-4-(1-(p-tolyl)ethyl)aniline (abbreviated DMPA). The presence of quartet and doublet peaks in NMR and a single chromatogram in HPLC verified that the final product DMPA, prepared from the synthesis reactions, had no major impurities...
June 7, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/28648994/dynamic-and-differential-expression-of-the-gonadal-aromatase-during-the-process-of-sexual-differentiation-in-a-novel-transgenic-cyp19a1a-egfp-zebrafish-line
#20
Nathalie Hinfray, Frédéric Sohm, Morgane Caulier, Edith Chadili, Benjamin Piccini, Camille Torchy, Jean-Marc Porcher, Yann Guiguen, François Brion
In zebrafish, there exists a clear need for new tools to study sex differentiation dynamic and its perturbation by endocrine disrupting chemicals. In this context, we developed and characterized a novel transgenic zebrafish line expressing green fluorescent protein (GFP) under the control of the zebrafish cyp19a1a (gonadal aromatase) promoter. In most gonochoristic fish species including zebrafish, cyp19a1a, the enzyme responsible for the synthesis of estrogens, has been shown to play a critical role in the processes of reproduction and sexual differentiation...
June 22, 2017: General and Comparative Endocrinology
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