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https://www.readbyqxmd.com/read/28103588/quantifying-kinetics-from-time-series-of-single-molecule-fret-efficiency-histograms
#1
Stephan Benke, Daniel Nettels, Hagen Hofmann, Benjamin Schuler
Single-molecule fluorescence spectroscopy is a powerful approach for probing biomolecular structure and dynamics, including protein folding. For the investigation of nonequilibrium kinetics, F��rster resonance energy transfer (FRET) in combination with confocal multiparameter detection has proven particularly versatile, owing to the large number of observables and the broad range of accessible timescales, especially in combination with rapid microfluidic mixing. However, a comprehensive kinetic analysis of the resulting time series of transfer efficiency histograms and complementary observables can be challenging owing to the complexity of the data...
January 19, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28103316/alpha-melanocyte-stimulating-hormone-protects-against-cytokine-induced-barrier-damage-in-caco-2-intestinal-epithelial-monolayers
#2
Judit Váradi, András Harazin, Ferenc Fenyvesi, Katalin Réti-Nagy, Péter Gogolák, György Vámosi, Ildikó Bácskay, Pálma Fehér, Zoltán Ujhelyi, Gábor Vasvári, Eszter Róka, David Haines, Mária A Deli, Miklós Vecsernyés
Alpha-melanocyte-stimulating hormone (α-MSH) is a potent anti-inflammatory peptide with cytoprotective effect in various tissues. The present investigation demonstrates the ability of α-MSH to interact with intestinal epithelial cell monolayers and mitigate inflammatory processes of the epithelial barrier. The protective effect of α-MSH was studied on Caco-2 human intestinal epithelial monolayers, which were disrupted by exposure to tumor necrosis factor-α and interleukin-1β. The barrier integrity was assessed by measuring transepithelial electric resistance (TEER) and permeability for marker molecules...
2017: PloS One
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
#3
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/28103286/mirnas-do-not-regulate-circadian-protein-synthesis-in-the-dinoflagellate-lingulodinium-polyedrum
#4
Steve Dagenais-Bellefeuille, Mathieu Beauchemin, David Morse
Dinoflagellates have been shown to express miRNA by bioinformatics and RNA blot (Northern) analyses. However, it is not yet known if miRNAs are able to alter gene expression in this class of organisms. We have assessed the possibility that miRNA may mediate circadian regulation of gene expression in the dinoflagellate Lingulodinium polyedrum using the Luciferin Binding Protein (LBP) as a specific example. LBP is a good candidate for regulation by miRNA since mRNA levels are constant over the daily cycle while protein synthesis is restricted by the circadian clock to a period of several hours at the start of the night phase...
2017: PloS One
https://www.readbyqxmd.com/read/28103015/antimicrobial-activity-ame-resistance-and-translation-inhibition-studies-of-anthraquinone-neomycin-conjugates
#5
Natalya N Degtyareva, Changjun Gong, Sandra Story, Nathanael Levinson, Adegboyega K Oyelere, Keith D Green, Sylvie Garneau-Tsodikova, Dev Priya Arya
The antibacterial effects of aminoglycosides are based on their association with the A-site of bacterial ribosomal RNA and interference with the translational process in the bacterial cell, causing cell death. The clinical use of aminoglycosides is complicated by resistance and side effects, some of which arise from their interactions with the human mitochondrial 12S rRNA and its deafness-associated mutations, C1494U and A1555G. We report a rapid assay that allows screening of aminoglycoside compounds to these classes of rRNAs...
January 19, 2017: ACS Infectious Diseases
https://www.readbyqxmd.com/read/28103010/a-fluorescent-hsp90-probe-demonstrates-the-unique-association-between-extracellular-hsp90-and-malignancy-in-vivo
#6
Lauren B Crowe, Philip F Hughes, David A Alcorta, Takuya Osada, Aaron P Smith, Juliane Totzke, David R Loiselle, Isaac D Lutz, Madhusudhana Gargesha, Debasish Roy, Jose Roques, David Darr, H Kim Lyerly, Neil L Spector, Timothy Aj Haystead
Extracellular expression of heat shock protein 90 (eHsp90) by tumor cells is correlated with malignancy. Development of small molecule probes that can detect eHsp90 in vivo may therefore have utility in the early detection of malignancy. We synthesized a cell impermeable far-red fluorophore-tagged Hsp90 inhibitor to target eHsp90 in vivo. High resolution confocal and lattice light sheet microscopy show that probe-bound eHsp90 accumulates in punctate structures on the plasma membrane of breast tumor cells and is actively internalized...
January 19, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28102753/chloroplastic-iron-sulfur-scaffold-protein-nfu3-is-essential-to-overall-plant-fitness
#7
Krishna Nath, James P O'Donnell, Yan Lu
A previous study showed that Nitrogen-Fixing-subunit-U-type protein NFU3 may act an iron-sulfur scaffold protein in the assembly and transfer of 4Fe-4S and 3Fe-4S clusters in the chloroplast. Examples of 4Fe-4S and 3Fe-4S-requiring proteins and complexes include Photosystem I (PSI), NAD(P)H dehydrogenase, and ferredoxin-dependent glutamine oxoglutarate aminotransferases. In this paper, the authors provided additional evidence for the role of NFU3 in 4Fe-4S and 3Fe-4S cluster assembly and transfer, as well as its role in overall plant fitness...
January 19, 2017: Plant Signaling & Behavior
https://www.readbyqxmd.com/read/28102396/zinc-sparks-induce-physiochemical-changes-in-the-egg-zona-pellucida-that-prevent-polyspermy
#8
Emily L Que, Francesca E Duncan, Amanda R Bayer, Steven J Philips, Eric W Roth, Reiner Bleher, Sophie C Gleber, Stefan Vogt, Teresa K Woodruff, Thomas V O'Halloran
During fertilization or chemically-induced egg activation, the mouse egg releases billions of zinc atoms in brief bursts known as 'zinc sparks.' The zona pellucida (ZP), a glycoprotein matrix surrounding the egg, is the first structure zinc ions encounter as they diffuse away from the plasma membrane. Following fertilization, the ZP undergoes changes described as 'hardening', which prevent multiple sperm from fertilizing the egg and thereby establish a block to polyspermy. A major event in zona hardening is cleavage of ZP2 proteins by ovastacin; however, the overall physiochemical changes contributing to zona hardening are not well understood...
January 19, 2017: Integrative Biology: Quantitative Biosciences From Nano to Macro
https://www.readbyqxmd.com/read/28102317/time-resolved-neutron-scattering-provides-new-insight-into-protein-substrate-processing-by-a-aaa-unfoldase
#9
Ziad Ibrahim, Anne Martel, Martine Moulin, Henry S Kim, Michael Härtlein, Bruno Franzetti, Frank Gabel
We present a combination of small-angle neutron scattering, deuterium labelling and contrast variation, temperature activation and fluorescence spectroscopy as a novel approach to obtain time-resolved, structural data individually from macromolecular complexes and their substrates during active biochemical reactions. The approach allowed us to monitor the mechanical unfolding of a green fluorescent protein model substrate by the archaeal AAA+ PAN unfoldase on the sub-minute time scale. Concomitant with the unfolding of its substrate, the PAN complex underwent an energy-dependent transition from a relaxed to a contracted conformation, followed by a slower expansion to its initial state at the end of the reaction...
January 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28102176/synthesis-of-magnetic-cytosine-imprinted-chitosan-nanoparticles
#10
Mei-Hwa Lee, Arti Ahluwalia, Jian-Zhou Chen, Neng-Lang Shih, Hung-Yin Lin
Molecularly imprinted polymer nanoparticles incorporating magnetic nanoparticles (MNPs) have been investigated for their selective adsorption properties. Here we describe the synthesis and characterization of magnetic cytosine-imprinted chitosan nanoparticles (CIPs) for gene delivery. In particular, CIPs carrying the mammalian expression plasmid of enhanced green fluorescent protein were prepared by the co-precipitation of MNPs, chitosan and a template nucleobase (cytosine). The results show that the selective reabsorption of cytosine to magnetic CIPs was at least double that of non-imprinted polymers and other nucleobases (such as adenine and thymine)...
January 19, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28102011/local-myogenic-pulp-derived-cell-injection-enhances-craniofacial-muscle-regeneration-in-vivo
#11
J E Jung, M J Song, S Shin, Y J Choi, K H Kim, C J Chung
OBJECTIVES: To enhance myogenic differentiation in pulp cells isolated from extracted premolars by epigenetic modification using a DNA demethylation agent, 5-aza-2'-deoxycytidine (5-Aza), and to evaluate the potent stimulatory effect of 5-Aza-treated pulp cell injection for craniofacial muscle regeneration in vivo. SETTING AND SAMPLE POPULATION: Pulp cells were isolated from premolars extracted for orthodontic purposes from four adults (age range, 18-22.1 years)...
February 2017: Orthodontics & Craniofacial Research
https://www.readbyqxmd.com/read/28101532/overexpression-of-brain-derived-neurotrophic-factor-protects-large-retinal-ganglion-cells-after-optic-nerve-crush-in-mice
#12
Liang Feng, Zhen Puyang, Hui Chen, Peiji Liang, John B Troy, Xiaorong Liu
Brain-derived neurotrophic factor (BDNF), a neurotrophin essential for neuron survival and function, plays an important role in neuroprotection during neurodegenerative diseases. In this study, we examined whether a modest increase of retinal BDNF promotes retinal ganglion cell (RGC) survival after acute injury of the optic nerve in mice. We adopted an inducible Cre-recombinase transgenic system to up-regulate BDNF in the mouse retina and then examined RGC survival after optic nerve crush by in vivo imaging...
January 2017: ENeuro
https://www.readbyqxmd.com/read/28101420/compact-diode-laser-source-for-multiphoton-biological-imaging
#13
Robert D Niederriter, Baris N Ozbay, Gregory L Futia, Emily A Gibson, Juliet T Gopinath
We demonstrate a compact, pulsed diode laser source suitable for multiphoton microscopy of biological samples. The center wavelength is 976 nm, near the peak of the two-photon cross section of common fluorescent markers such as genetically encoded green and yellow fluorescent proteins. The laser repetition rate is electrically tunable between 66.67 kHz and 10 MHz, with 2.3 ps pulse duration and peak powers >1 kW. The laser components are fiber-coupled and scalable to a compact package. We demonstrate >600 μm depth penetration in brain tissue, limited by laser power...
January 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28101336/regulatory-elements-and-transcriptional-control-of-chicken-vasa-homologue-cvh-promoter-in-chicken-primordial-germ-cells
#14
So Dam Jin, Bo Ram Lee, Young Sun Hwang, Hong Jo Lee, Jong Seop Rim, Jae Yong Han
BACKGROUND: Primordial germ cells (PGCs), the precursors of functional gametes, have distinct characteristics and exhibit several unique molecular mechanisms to maintain pluripotency and germness in comparison to somatic cells. They express germ cell-specific RNA binding proteins (RBPs) by modulating tissue-specific cis- and trans-regulatory elements. Studies on gene structures of chicken vasa homologue (CVH), a chicken RNA binding protein, involved in temporal and spatial regulation are thus important not only for understanding the molecular mechanisms that regulate germ cell fate, but also for practical applications of primordial germ cells...
2017: Journal of Animal Science and Biotechnology
https://www.readbyqxmd.com/read/28101201/diosmetin-inhibits-cell-proliferation-and-induces-apoptosis-by-regulating-autophagy-via-the-mammalian-target-of-rapamycin-pathway-in-hepatocellular-carcinoma-hepg2-cells
#15
Jie Liu, Hao Ren, Bin Liu, Qingyu Zhang, Mingyi Li, Runzhi Zhu
Hepatocellular carcinoma (HCC), which is a type of malignant tumor, is the fifth most common cancer in men and ninth in women worldwide. The aim of the present study was to investigate the antitumor effect of diosmetin (DIOS) in hepatocellular carcinoma HepG2 cells. The proliferation, apoptosis and autophagy rates of HepG2 cells were measured following treatment with DIOS. The effects of DIOS treatment on HepG2 cell proliferation and apoptosis rates were analyzed using MTT assays and Annexin V staining, respectively...
December 2016: Oncology Letters
https://www.readbyqxmd.com/read/28101198/elevated-expression-of-nrf2-mediates-multidrug-resistance-in-cd133-head-and-neck-squamous-cell-carcinoma-stem-cells
#16
Bao-Cai Lu, Jing Li, Wen-Fa Yu, Guo-Zheng Zhang, Hui-Min Wang, Hui-Min Ma
Enhanced expression of the ATP-binding cassette (ABC) transporter protein ABC sub-family G member 2 (ABCG2) in cancer stem cells (CSCs) plays a major role in chemotherapeutic drug efflux, which results in therapy failure and tumor relapse. In addition to downregulating apoptosis in CSCs, it has been reported that the transcriptional upregulation of the redox sensing factor Nrf2 is involved in the upregulation of ABCG2 expression and consequent chemoresistance. The current study investigated the presence of cancer stem-like side population (SP) cells from head and neck squamous cell carcinoma (HNSCC) samples, and evaluated the Nrf2 expression profile and multidrug resistance properties of HNSCC stem cells...
December 2016: Oncology Letters
https://www.readbyqxmd.com/read/28101152/the-association-between-herg-gene-expression-and-cardiac-arrhythmia-disease-in-children
#17
Mingyu Fu, Chunli Li, Qingwen Wang, Yan Wang, Xinjiang An
We explored the possible link between the expression of HERG gene and cardiomyopathy in children. From April 2013 to April 2015, 73 children with cardiac arrhythmia who were treated were enrolled in the present study to serve as the observation group. At the same time, 76 normal individuals were also enrolled as the control group. HERG expression level in the observation group was compared with the control group. To determine the level of HERG gene expression we used fluorescent directional PCR, enzyme immunoassay and western blot analysis...
December 2016: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/28100205/a-fast-and-effective-determination-of-the-biodistribution-and-subcellular-localization-of-fluorescent-immunoliposomes-in-freshly-excised-animal-organs
#18
Felista L Tansi, Ronny Rüger, Ansgar M Kollmeier, Claudia Böhm, Roland E Kontermann, Ulf K Teichgraeber, Alfred Fahr, Ingrid Hilger
BACKGROUND: Preclinical research implementing fluorescence-based approaches is inevitable for drug discovery and technology. For example, a variety of contrast agents developed for biomedical imaging are usually evaluated in cell systems and animal models based on their conjugation to fluorescent dyes. Biodistribution studies of excised organs are often performed by macroscopic imaging, whereas the subcellular localization though vital, is often neglected or further validated by histological procedures...
January 18, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28099027/inactivation-of-soybean-trypsin-inhibitor-by-epigallocatechin-gallate-stopped-flow-fluorescence-thermodynamics-and-docking-studies
#19
Chun Liu, Fenfen Cheng, Xiaoquan Yang
Tea is one of the most widely daily consumed beverages all over the world, and it is usually consumed with milk and/or soy milk. However, very few researches have studied the interactions between tea polyphenols (TPs) and soy milk proteins as compared with milk proteins. Here, we reported that epigallocatechin gallate (EGCG), a major component of TPs, can effectively inhibit the inhibitory activity of Kunitz trypsin inhibitor (KTI, a major antinutrient in soy milk). The mechanism of inactivation of KTI by EGCG was investigated by stopped-flow/fluorescence, thermodynamics, and docking studies...
January 18, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28098972/hepta-mutant-staphylococcus-aureus-sortase-a-srta7m-as-a-tool-for-in-vivo-protein-labeling-in-caenorhabditis-elegans
#20
Qin Wu, Hidde L Ploegh, Matthias C Truttmann
In vivo protein ligation is of emerging interest as a means of endowing proteins with new properties in a controlled fashion. Tools to site-specifically and covalently modify proteins with small molecules, peptides, or other proteins in living cells are few and far between. Here, we describe the development of a Staphylococcus aureus sortase (SrtA)-based protein ligation approach for site-specific conjugation of fluorescent dyes and ubiquitin (Ub) to modify proteins in Caenorhabditis elegans. Hepta-mutant SrtA (SrtA7m) expressed in C...
January 18, 2017: ACS Chemical Biology
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