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Eveline Brodl, Andreas Winkler, Peter Macheroux
Bioluminescence refers to the production of light by living organisms. Bioluminescent bacteria with a variety of bioluminescence emission characteristics have been identified in Vibrionaceae , Shewanellaceae and Enterobacteriaceae . Bioluminescent bacteria are mainly found in marine habitats and they are either free-floating, sessile or have specialized to live in symbiosis with other marine organisms. On the molecular level, bioluminescence is enabled by a cascade of chemical reactions catalyzed by enzymes encoded by the lux operon with the gene order luxCDABEG ...
2018: Computational and Structural Biotechnology Journal
Ulrike Binder, Maria Isabel Navarro-Mendoza, Verena Naschberger, Ingo Bauer, Francisco E Nicolas, Johannes D Pallua, Cornelia Lass-Flörl, Victoriano Garre
Invasive fungal infections caused by Mucorales (mucormycosis) have increased worldwide. These life-threatening infections affect mainly, but not exclusively, immunocompromised patients, and are characterized by rapid progression, severe tissue damage and an unacceptably high rate of mortality. Still, little is known about this disease and its successful therapy. New tools to understand mucormycosis and a screening method for novel antimycotics are required. Bioluminescent imaging is a powerful tool for in vitro and in vivo approaches...
December 7, 2018: Genes
Giulia Brigante, Laura Riccetti, Clara Lazzaretti, Laura Rofrano, Samantha Sperduti, Francesco Potì, Chiara Diazzi, Flavia Prodam, Giovanni Guaraldi, Andrea G Lania, Vincenzo Rochira, Livio Casarini
Growth Hormone (GH) deficiency is frequent in HIV-infected patients treated with antiretroviral therapy. We treated GH3 cells with antiretrovirals (nevirapine, ritonavir or abacavir sulfate; 100 pM-1 mM range), after transfection with human growth hormone releasing hormone (GHRH) receptor cDNA. Cells viability, intracellular cAMP, phosphorylation of CREB and calcium increase, GH production and secretion were evaluated both in basal condition and after GHRH, using MTT, bioluminescence resonance energy transfer, western blotting and ELISA...
December 10, 2018: Molecular and Cellular Endocrinology
Hung-I Lin, Kanchan Yadav, Kun-Ching Shen, Golam Haider, Pradip Kumar Roy, Monika Kataria, Ting-Jia Chang, Yao-Hsuan Li, Tai-Yuan Lin, Yit-Tsong Chen, Yang-Fang Chen
Plasmonic material has been emerged with multi-functionalities for its remarkable tailoring light emission, reshaping density of states (DOS) and focusing subwavelength light. However, restricted by its propagation loss and narrowband resonance in nature, it is a challenge issue for plasmonic material to provide a broadband DOS to advance its application. Here, we develop a novel nanoscale core-shell hyperbolic structure that possesses a remarkably coupling effect inside the multishell nanoscale composite owing to a higher DOS and longer time of collective oscillations of the electrons than the plasmonic-based pure metal nanoparticles...
December 13, 2018: ACS Applied Materials & Interfaces
Roderick Adrian Slavcev, Chi Hong Sum, Jesse St Jean, Haein Huh, Nafiseh Nafissi
Fluorescent-based visualization techniques have long been used to monitor biological activity. This chapter explores the delivery of reporter genes as a means to assay and track activity in biological systems. Bioluminescence is the production of light due to biochemical processes. By encoding genes for bioluminescence, biological processes can be visualized based on gene expression. This chapter also discusses the primary applications of bioluminescence as seen through bioluminescent imaging techniques, flow cytometry, and PCR-based methods of gene detection...
2018: Experientia. Supplementum
Edwin Chang, Chirag B Patel, Christoph Pohling, Caroline Young, Jonathan Song, Thomas Anthony Flores, Yitian Zeng, Lydia-Marie Joubert, Hamed Arami, Arutselvan Natarajan, Robert Sinclair, Sanjiv S Gambhir
Glioblastoma is the most common yet most lethal of primary brain cancers with a one-year post-diagnosis survival rate of 65% and a five-year survival rate of barely 5%. Recently the U.S. Food and Drug Administration approved a novel fourth approach (in addition to surgery, radiation therapy, and chemotherapy) to treating glioblastoma; namely, tumor treating fields (TTFields). TTFields involves the delivery of alternating electric fields to the tumor but its mechanisms of action are not fully understood. Current theories involve TTFields disrupting mitosis due to interference with proper mitotic spindle assembly...
2018: Cell Death Discovery
Isao T Tokuda, Daisuke Ono, Sato Honma, Ken-Ichi Honma, Hanspeter Herzel
Circadian clocks are autonomous oscillators driving daily rhythms in physiology and behavior. In mammals, a network of coupled neurons in the suprachiasmatic nucleus (SCN) is entrained to environmental light-dark cycles and orchestrates the timing of peripheral organs. In each neuron, transcriptional feedbacks generate noisy oscillations. Coupling mediated by neuropeptides such as VIP and AVP lends precision and robustness to circadian rhythms. The detailed coupling mechanisms between SCN neurons are debated...
December 10, 2018: PLoS Computational Biology
Maria J L Kracht, Eelco J P de Koning, Rob C Hoeben, Bart O Roep, Arnaud Zaldumbide
During type 1 diabetes development, cells in the islets of Langerhans engage adaptive mechanisms in response to inflammatory signals to cope with stress, to restore cellular homeostasis, and to preserve cell function. Disruption of these mechanisms may induce the formation of a repertoire of stress-induced neoantigens, which are critical in the loss of tolerance to beta cells and the development of autoimmunity. While multiple lines of evidence argue for a critical role of the endoplasmic reticulum in these processes, the lack of tools to specifically monitor beta cell stress hampers the development of therapeutic interventions focusing on maintaining endoplasmic reticulum homeostasis...
December 10, 2018: Scientific Reports
Haoyu Sun, Min Zheng, Jinyuan Song, Shengyou Huang, Yongzheng Pan, Ruochong Gong, Zhifen Lin
Hormesis is a dose-response relationship phenomenon characterized by low-dose stimulation and high-dose inhibition. Although hormetic phenomena have been reported in broadly ranging biological areas, there is still no unified mechanism of hormesis. Investigating multiple-species hormesis of one compound and then exploring the possible mechanism may be an effective approach to clarify the reason for the occurrence of hormetic phenomena in a broad range of organisms. In this study, indole was selected as the test chemical due to the broad biological and hormetic effects of indole compounds...
December 3, 2018: Science of the Total Environment
You-Cheng Hseu, Yi-Chun Lin, Peramaiyan Rajendran, Varadharajan Thigarajan, Dony Chacko Mathew, Kai-Yuan Lin, Tzong-Der Way, Jiunn-Wang Liao, Hsin-Ling Yang
Antrodia salmonea (AS), a fungus that is indigenous to Taiwan has been well known for its anti-cancer properties. We investigated the anti-metastatic and anti-epithelial-mesenchymal transition (EMT) properties of AS in TNBC cells. To determine their EMT and metastasis levels, in vitro wound healing, wound invasion, Western blotting, RT-PCR, luciferase activity and immunofluorescence assays were performed, while the in vivo anti-metastatic efficacy of AS was evaluated in BALB/c-nu mice through bioluminescence imaging, HE staining, and immunohistochemical staining...
December 7, 2018: Food and Chemical Toxicology
Kouki K Touhara, Roderick MacKinnon
Stimulated muscarinic acetylcholine receptors (M2Rs) release Gβγ subunits, which slow heart rate by activating a G protein-gated K+ channel (GIRK). Stimulated β2 adrenergic receptors (β2ARs) also release Gβγ subunits, but GIRK is not activated. This study addresses the mechanism underlying this specificity of GIRK activation by M2Rs. K+ currents and bioluminescence resonance energy transfer between labelled G proteins and GIRK show that M2Rs catalyze Gβγ subunit release at higher rates than β2ARs, generating higher Gβγ concentrations that activate GIRK and regulate other targets of Gβγ...
December 10, 2018: ELife
Yan Ni, Remco Arts, Maarten Merkx
Bioluminescent sensor proteins provide attractive tools for applications ranging from in vivo imaging to point-of-care testing. Here we introduce a new class of ratiometric bioluminescent sensor proteins that do not rely on direct modulation of BRET efficiency, but are based on competitive intramolecular complementation of split NanoLuc luciferase. Proof of concept for the feasibility of this sensor principle was provided by developing a blue-red light emitting sensor protein for the detection of anti-HIV1-p17 antibodies with a 500% change in emission ratio and a Kd of 10 pM...
December 11, 2018: ACS Sensors
Patrick Hardinge, Guy Kiddle, Laurence Tisi, James A H Murray
Loop-mediated amplification (LAMP) has been widely used to amplify and hence detect nucleic acid target sequences from various pathogens, viruses and genetic modifications. Two distinct types of primer are required for LAMP; hairpin-forming LAMP and displacement. High specificity arises from this use of multiple primers, but without optimal conditions for LAMP, sensitivity can be poor. We confirm here the importance of LAMP primer design, concentrations and ratios for efficient LAMP amplification. We further show that displacement primers are non-essential to the LAMP reaction at certain concentrations providing accelerating loop primers are present...
December 4, 2018: Scientific Reports
Vadim R Viviani, Gabriele V M Gabriel, Vanessa R Bevilaqua, A F Simões, T Hirano, P S Lopes-de-Oliveira
Firefly luciferases produce yellow-green light under physiological and alkaline conditions, however at acidic pH, higher temperatures or in the presence of heavy metals the color changes to red, a property called pH-sensitivity. Despite many decades of studies, the proton and metal binding sites responsible for pH-sensitivity remain enigmatic. Previously we suggested that the salt bridge E311/R337 keeps a closed conformation of the luciferin phenolate binding site. Here we further investigated the effect of this salt bridge and mutations of the neighbor residues H310 and E/N354, on metal and pH-sensitivity of firefly luciferases emitting distinct bioluminescence colors (Cratomorphus distinctus: 548 nm; Macrolampis sp2: 569 nm)...
December 4, 2018: Scientific Reports
Yoon Namkung, Christian LeGouill, Sahil Kumar, Yubo Cao, Larissa B Teixeira, Viktoriya Lukasheva, Jenna Giubilaro, Sarah C Simões, Jean-Michel Longpré, Dominic Devost, Terence E Hébert, Graciela Piñeyro, Richard Leduc, Claudio M Costa-Neto, Michel Bouvier, Stéphane A Laporte
G protein-coupled receptors (GPCRs) are important therapeutic targets that exhibit functional selectivity (biased signaling), in which different ligands or receptor variants elicit distinct downstream signaling. Understanding all the signaling events and biases that contribute to both the beneficial and adverse effects of GPCR stimulation by given ligands is important for drug discovery. Here, we report the design, validation, and use of pathway-selective bioluminescence resonance energy transfer (BRET) biosensors that monitor the engagement and activation of signaling effectors downstream of G proteins, including protein kinase C (PKC), phospholipase C (PLC), p63RhoGEF, and Rho...
December 4, 2018: Science Signaling
Marc Prante, Christian Ude, Miriam Große, Lukas Raddatz, Ulrich Krings, Gernot John, Shimshon Belkin, Thomas Scheper
Buried explosive material, e.g., landmines, represent a severe issue for human safety all over the world. Most explosives consist of environmentally hazardous chemicals like 2,4,6-trinitrotoluene (TNT), carcinogenic 2,4-dinitrotoluene (2,4-DNT) and related compounds. Vapors leaking from buried landmines offer a detection marker for landmines, presenting an option to detect landmines without relying on metal detection. 2,4-Dinitrotoluene (DNT), an impurity and byproduct of common TNT synthesis, is a feasible detection marker since it is extremely volatile...
December 3, 2018: Sensors
Yuji Kaneko, Julian P Tuazon, Xunming Ji, Cesario V Borlongan
BACKGROUND/AIMS: The endogenous neurotrophic peptides pituitary adenylate cyclase-activating polypeptides (PACAP-27/38) protect against stroke, but the molecular mechanism remains unknown. METHODS: Primary rat neural cells were exposed to PACAP-27 or PACAP-38 before induction of experimental acute ischemic stroke via oxygen-glucose deprivation-reperfusion (OGD/R) injury. To reveal PACAP's role in neuroprotection, we employed fluorescent live/dead cell viability and caspase 3 assays, optical densitometry of mitochondrial dehydrogenase and cell growth, glutathione disulfide luciferase activity, ELISA for high mobility group box1 extracellular concentration, ATP bioluminescence, Western blot analysis of PACAP, NMDA subunits, apoptosis regulator Bcl-2, social interaction hormone oxytocin, and trophic factor BDNF, and immunocytochemical analysis of PACAP...
December 4, 2018: Cellular Physiology and Biochemistry
Arkadiy A Bazhin, Marc Chambon, Jonathan Vesin, Julien Bortoli, James W Collins, Gerardo Turcatti, Chieh Jason Chou, Elena A Goun
Aminopeptidases, such as dipeptidyl peptidase-4 (DPP-4, CD26), are potent therapeutic targets for pharmacological interventions because they play key roles in many important pathological pathways. To analyze aminopeptidase activity in vitro (including high throughput screening [HTS]), in vivo, and ex vivo, we developed a highly sensitive and quantitative bioluminescence-based readout method. We successfully applied this method to screening drugs with potential DPP-4 inhibitory activity. Using this method, we found that cancer drug mitoxantrone possesses significant DPP-4 inhibitory activity both in vitro and in vivo...
December 4, 2018: Analytical Chemistry
Ioan A Lina, Wataru Ishida, Jason A Liauw, Sheng-Fu L Lo, Benjamin D Elder, Alexander Perdomo-Pantoja, Debebe Theodros, Timothy F Witham, Christina Holmes
PURPOSE: Bone marrow aspirate has been successfully used alongside a variety of grafting materials to clinically augment spinal fusion. However, little is known about the fate of these transplanted cells. Herein, we develop a novel murine model for the in vivo monitoring of implanted bone marrow cells (BMCs) following spinal fusion. METHODS: A clinical-grade scaffold was implanted into immune-intact mice undergoing spinal fusion with or without freshly isolated BMCs from either transgenic mice which constitutively express the firefly luciferase gene or syngeneic controls...
December 3, 2018: European Spine Journal
Cátia Vieira, Ana T P C Gomes, Mariana Q Mesquita, Nuno M M Moura, M Graça P M S Neves, M Amparo F Faustino, Adelaide Almeida
Antimicrobial photodynamic therapy (aPDT) is gaining a special importance as an effective approach against multidrug-resistant strains responsible of fatal infections. The addition of potassium iodide (KI), a non-toxic salt, is recognized to increase the aPDT efficiency of some photosensitizers (PSs) on a broad-spectrum of microorganisms. As the reported cases only refer positive aPDT potentiation results, in this work we selected a broad range of porphyrinic and non-porphyrinic PSs in order to gain a more comprehensive knowledge about this aPDT potentiation by KI...
2018: Frontiers in Microbiology
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