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https://www.readbyqxmd.com/read/27919754/neutralization-takes-precedence-over-igg-or-iga-isotype-related-functions-in-mucosal-hiv-1-antibody-mediated-protection
#1
Rena D Astronomo, Sampa Santra, Lamar Ballweber-Fleming, Katharine G Westerberg, Linh Mach, Tiffany Hensley-McBain, Laura Sutherland, Benjamin Mildenberg, Georgeanna Morton, Nicole L Yates, Gregory J Mize, Justin Pollara, Florian Hladik, Christina Ochsenbauer, Thomas N Denny, Ranjit Warrier, Supachai Rerks-Ngarm, Punnee Pitisuttithum, Sorachai Nitayapan, Jaranit Kaewkungwal, Guido Ferrari, George M Shaw, Shi-Mao Xia, Hua-Xin Liao, David C Montefiori, Georgia D Tomaras, Barton F Haynes, M Juliana McElrath
HIV-1 infection occurs primarily through mucosal transmission. Application of biologically relevant mucosal models can advance understanding of the functional properties of antibodies that mediate HIV protection, thereby guiding antibody-based vaccine development. Here, we employed a human ex vivo vaginal HIV-1 infection model and a rhesus macaque in vivo intrarectal SHIV challenge model to probe the protective capacity of monoclonal broadly-neutralizing (bnAb) and non-neutralizing Abs (nnAbs) that were functionally modified by isotype switching...
November 21, 2016: EBioMedicine
https://www.readbyqxmd.com/read/27853830/exploiting-nanoluc-luciferase-for-smartphone-based-bioluminescence-cell-biosensor-for-anti-inflammatory-activity-and-toxicity
#2
Luca Cevenini, Maria Maddalena Calabretta, Antonia Lopreside, Giuseppe Tarantino, Annalisa Tassoni, Maura Ferri, Aldo Roda, Elisa Michelini
The availability of smartphones with high-performance digital image sensors and processing power has completely reshaped the landscape of point-of-need analysis. Thanks to the high maturity level of reporter gene technology and the availability of several bioluminescent proteins with improved features, we were able to develop a bioluminescence smartphone-based biosensing platform exploiting the highly sensitive NanoLuc luciferase as reporter. A 3D-printed smartphone-integrated cell biosensor based on genetically engineered Hek293T cells was developed...
December 2016: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/27786307/coupling-optogenetic-stimulation-with-nanoluc-based-luminescence-bret-ca-sensing
#3
Jie Yang, Derrick Cumberbatch, Samuel Centanni, Shu-Qun Shi, Danny Winder, Donna Webb, Carl Hirschie Johnson
Optogenetic techniques allow intracellular manipulation of Ca(++) by illumination of light-absorbing probe molecules such as channelrhodopsins and melanopsins. The consequences of optogenetic stimulation would optimally be recorded by non-invasive optical methods. However, most current optical methods for monitoring Ca(++) levels are based on fluorescence excitation that can cause unwanted stimulation of the optogenetic probe and other undesirable effects such as tissue autofluorescence. Luminescence is an alternate optical technology that avoids the problems associated with fluorescence...
October 27, 2016: Nature Communications
https://www.readbyqxmd.com/read/27779402/detection-and-activity-profiling-of-synthetic-cannabinoids-and-metabolites-with-a-newly-developed-bio-assay
#4
Annelies Cannaert, Jolien Storme, Florian Franz, Volker Auwärter, Christophe P Stove
Synthetic cannabinoids (SCs) are the largest group of compounds currently monitored in Europe by the EU Early Warning System on new psychoactive substances. Emerging recreational use of these products has led to multiple cases of adverse health effects and even death. In contrast to marijuana, where Δ9-tetrahydrocannabinol (Δ9THC) is metabolized to only one major active metabolite, it has been reported that several major phase I metabolites of SCs remain biologically active, exerting cannabinoid (CB) receptor affinity, potency and efficacy greater than Δ9THC...
October 25, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/27687581/sod1-dimerization-monitoring-using-a-novel-split-nanoluc-nanobit
#5
Kentaro Oh-Hashi, Yoko Hirata, Kazutoshi Kiuchi
In the present study, we applied a highly sensitive NanoLuc-based technology to understand the status of superoxide dismutase 1 (SOD1) within mammalian cells. Two fragments of NanoLuc (NanoBit), large N-terminal and small C-terminal regions, were fused with wild-type (wt) and mutant human SOD1 (hSOD1) genes and transfected into cells. Luciferase activity through NanoBit assembly was only detected in NanoBit-tagged wtSOD1-expressing cells. Furthermore, the developed NanoLuc system was used to investigate the role of protein-protein interactions in the pathogenesis of amyotrophic lateral sclerosis (ALS)...
October 2016: Cell Biochemistry and Function
https://www.readbyqxmd.com/read/27624517/the-use-of-a-novel-nanoluc-based-reporter-phage-for-the-detection-of-escherichia-coli-o157-h7
#6
Dandan Zhang, Claudia P Coronel-Aguilera, Patricia L Romero, Lynda Perry, Udit Minocha, Carla Rosenfield, Andrew G Gehring, George C Paoli, Arun K Bhunia, Bruce Applegate
Rapid detection of the foodborne pathogen Escherichia coli O157:H7 is of vital importance for public health worldwide. Among detection methods, reporter phages represent unique and sensitive tools for the detection of E. coli O157:H7 from food as they are host-specific and able to differentiate live cells from dead ones. Upon infection, target bacteria become identifiable since reporter genes are expressed from the engineered phage genome. The E. coli O157:H7 bacteriophage ΦV10 was modified to express NanoLuc luciferase (Nluc) derived from the deep-sea shrimp Oplophorus gracilirostris...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27588207/use-of-a-new-proximity-assay-nanobret-to-investigate-the-ligand-binding-characteristics-of-three-fluorescent-ligands-to-the-human-%C3%AE-1-adrenoceptor-expressed-in-hek-293-cells
#7
Mark Soave, Leigh A Stoddart, Alastair Brown, Jeanette Woolard, Stephen J Hill
Previous research has indicated that allosteric interactions across the dimer interface of β 1-adrenoceptors may be responsible for a secondary low affinity binding conformation. Here we have investigated the potential for probe dependence, in the determination of antagonist pKi values at the human β 1-adenoceptor, which may result from such allosterism interactions. Three fluorescent β 1-adrenoceptor ligands were used to investigate this using bioluminescence energy transfer (BRET) between the receptor-bound fluorescent ligand and the N-terminal NanoLuc tag of a human β 1-adrenoceptor expressed in HEK 293 cells (NanoBRET)...
October 2016: Pharmacology Research & Perspectives
https://www.readbyqxmd.com/read/27547982/dual-readout-bret-fret-sensors-for-measuring-intracellular-zinc
#8
Stijn J A Aper, Pieterjan Dierickx, Maarten Merkx
Genetically encoded FRET-based sensor proteins have significantly contributed to our current understanding of the intracellular functions of Zn(2+). However, the external excitation required for these fluorescent sensors can give rise to photobleaching and phototoxicity during long-term imaging, limits applications that suffer from autofluorescence and light scattering, and is not compatible with light-sensitive cells. For these applications, sensor proteins based on Bioluminescence Resonance Energy Transfer (BRET) would provide an attractive alternative...
October 21, 2016: ACS Chemical Biology
https://www.readbyqxmd.com/read/27518620/using-%C3%AE-lactamase-and-nanoluc-luciferase-reporter-gene-assays-to-identify-inhibitors-of-the-hif-1-signaling-pathway
#9
Thai Khuc, Chia-Wen Amy Hsu, Srilatha Sakamuru, Menghang Xia
The hypoxia-inducible factor 1 (HIF-1) is a transcriptional factor involved in the regulation of oxygen within cellular environments. In hypoxic tissues or those with inadequate oxygen concentrations, activation of the HIF-1 transcription factor allows for subsequent activation of target gene expression implicated in cell survival. As a result, cells proliferate through formation of new blood vessels and expansion of vascular systems, providing necessary nourishment needed of cells. HIF-1 is also involved in the complex pathophysiology associated with cancer cells...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27515429/application-of-nanoluc-to-monitor-the-intrinsic-promoter-activity-of-grp78-using-the-crispr-cas9-system
#10
Kentaro Oh-Hashi, Eri Furuta, Junpei Norisada, Fumimasa Amaya, Yoko Hirata, Kazutoshi Kiuchi
In this study, we applied a highly sensitive small luciferase, NanoLuc, to establish a knock-in cell line using the CRISPR/Cas9 system and characterized the endogenous promoter activity of the glucose-regulated protein 78 (GRP78) gene. The N-terminal region of the human GRP78 gene was fused to the NanoLuc gene and aligned with the puromycin-resistant gene through the 2A peptide sequence and used as a knock-in vector. The selected cells responded to both pharmacological and genetic ER stress and show NanoLuc-based CRISPR/Cas9 system is a very useful tool to isolate gene-edited cells and to characterize the endogenous promoter activity for genes of interest...
October 2016: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
https://www.readbyqxmd.com/read/27424896/bioluminescent-ligand-receptor-binding-assays-for-protein-or-peptide-hormones
#11
Ya-Li Liu, Zhan-Yun Guo
Bioluminescence has been widely used in biomedical research due to its high sensitivity, low background, and broad linear range. In recent studies, we applied bioluminescence to ligand-receptor binding assays for some protein or peptide hormones based on a newly developed small monomeric Nanoluciferase (NanoLuc) reporter that has the so far brightest bioluminescence. The conventional ligand-receptor binding assays rely on radioligands that have drawbacks, such as radioactive hazards and short shelf lives. In contrast, the novel bioluminescent binding assays use the NanoLuc-based protein or peptide tracers that are safe, stable, and ultrasensitive...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27414797/novel-bioluminescent-binding-assays-for-ligand-receptor-interaction-studies-of-the-fibroblast-growth-factor-family
#12
Ge Song, Xiao-Xia Shao, Qing-Ping Wu, Zeng-Guang Xu, Ya-Li Liu, Zhan-Yun Guo
We recently developed novel bioluminescent binding assays for several protein/peptide hormones to study their interactions with receptors using the so far brightest NanoLuc reporter. To validate the novel bioluminescent binding assay using a variety of protein/peptide hormones, in the present work we applied it to the fibroblast growth factor (FGF) family using the prototype member FGF2 as an example. A fully active recombinant FGF2 retaining a unique exposed cysteine (Cys) residue was chemically conjugated with an engineered NanoLuc carrying a unique exposed Cys residue at the C-terminus via formation of an intermolecular disulfide linkage...
2016: PloS One
https://www.readbyqxmd.com/read/27392906/elucidation-of-a-novel-phenformin-derivative-on-glucose-deprived-stress-responses-in-ht-29-cells
#13
Kentaro Oh-Hashi, Nao Irie, Takayuki Sakai, Kensuke Okuda, Hideko Nagasawa, Yoko Hirata, Kazutoshi Kiuchi
Recently, we developed a variety of phenformin derivatives as selective antitumor agents. Based on previous findings, this study evaluated a promising compound, 2-(2-chlorophenyl)ethylbiguanide (2-Cl-Phen), on the basis of stress responses in the human colon cancer cell line HT-29 under a serum- and glucose-deprived condition. 2-Cl-Phen triggered morphological changes such as shrinkage and plasma membrane disintegration, as well as a decrease in mitochondrial activity and an increase in LDH leakage. To understand intracellular issues relating to 2-Cl-Phen, this study focused on the expression levels of ER stress-inducible genes and several oncogenic genes...
August 2016: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/27317001/bret-nanoluc-based-bioluminescence-resonance-energy-transfer-platform-to-monitor-protein-protein-interactions-in-live-cells
#14
Xiu-Lei Mo, Haian Fu
Bioluminescence resonance energy transfer (BRET) is a prominent biophysical technology for monitoring molecular interactions, and has been widely used to study protein-protein interactions (PPI) in live cells. This technology requires proteins of interest to be associated with an energy donor (i.e., luciferase) and an acceptor (e.g., fluorescent protein) molecule. Upon interaction of the proteins of interest, the donor and acceptor will be brought into close proximity and energy transfer of chemical reaction-induced luminescence to its corresponding acceptor will result in an increased emission at an acceptor-defined wavelength, generating the BRET signal...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27240196/a-bright-cyan-excitable-orange-fluorescent-protein-facilitates-dual-emission-microscopy-and-enhances-bioluminescence-imaging-in-vivo
#15
Jun Chu, Younghee Oh, Alex Sens, Niloufar Ataie, Hod Dana, John J Macklin, Tal Laviv, Erik S Welf, Kevin M Dean, Feijie Zhang, Benjamin B Kim, Clement Tran Tang, Michelle Hu, Michelle A Baird, Michael W Davidson, Mark A Kay, Reto Fiolka, Ryohei Yasuda, Douglas S Kim, Ho-Leung Ng, Michael Z Lin
Orange-red fluorescent proteins (FPs) are widely used in biomedical research for multiplexed epifluorescence microscopy with GFP-based probes, but their different excitation requirements make multiplexing with new advanced microscopy methods difficult. Separately, orange-red FPs are useful for deep-tissue imaging in mammals owing to the relative tissue transmissibility of orange-red light, but their dependence on illumination limits their sensitivity as reporters in deep tissues. Here we describe CyOFP1, a bright, engineered, orange-red FP that is excitable by cyan light...
July 2016: Nature Biotechnology
https://www.readbyqxmd.com/read/27157466/quantum-dot-nanoluc-bioluminescence-resonance-energy-transfer-enables-tumor-imaging-and-lymph-node-mapping-in-vivo
#16
Anyanee Kamkaew, Haiyan Sun, Christopher G England, Liang Cheng, Zhuang Liu, Weibo Cai
A small luciferase protein (Nluc) was conjugated to QDs as a bioluminescence resonance energy transfer (BRET) pair. The conjugate showed 76% BRET efficiency and lymph node mapping was successfully performed. The cRGD peptide was conjugated to QD-Nluc for tumor targeting. The self-illuminating QD-Nluc showed excellent energy transfer in a living system and offered an optimal tumor-to-background ratio (>85).
May 19, 2016: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/27102897/an-ultrasensitive-nanoluc-based-luminescence-system-for-monitoring-plasmodium-berghei-throughout-its-life-cycle
#17
Mariana De Niz, Rebecca R Stanway, Rahel Wacker, Derya Keller, Volker T Heussler
BACKGROUND: Bioluminescence imaging is widely used for cell-based assays and animal imaging studies, both in biomedical research and drug development. Its main advantages include its high-throughput applicability, affordability, high sensitivity, operational simplicity, and quantitative outputs. In malaria research, bioluminescence has been used for drug discovery in vivo and in vitro, exploring host-pathogen interactions, and studying multiple aspects of Plasmodium biology. While the number of fluorescent proteins available for imaging has undergone a great expansion over the last two decades, enabling simultaneous visualization of multiple molecular and cellular events, expansion of available luciferases has lagged behind...
2016: Malaria Journal
https://www.readbyqxmd.com/read/27045664/nanoluc-a-small-luciferase-is-brightening-up-the-field-of-bioluminescence
#18
Christopher G England, Emily B Ehlerding, Weibo Cai
The biomedical field has greatly benefited from the discovery of bioluminescent proteins. Currently, scientists employ bioluminescent systems for numerous biomedical applications, ranging from highly sensitive cellular assays to bioluminescence-based molecular imaging. Traditionally, these systems are based on Firefly and Renilla luciferases; however, the applicability of these enzymes is limited by their size, stability, and luminescence efficiency. NanoLuc (NLuc), a novel bioluminescence platform, offers several advantages over established systems, including enhanced stability, smaller size, and >150-fold increase in luminescence...
May 18, 2016: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/27020777/novel-bioluminescent-binding-assays-for-interaction-studies-of-protein-peptide-hormones-with-their-receptors
#19
REVIEW
Ya-Li Liu, Zhan-Yun Guo
Protein/peptide hormones are the largest group of endogenous signaling molecules and exert various biological functions by binding to specific cell membrane receptors. To study the interactions between these hormones and their receptors, quantitative ligand-receptor binding assays have been widely used for decades. However, the assays conventionally relied on the use of radioligands, which have some major drawbacks and can only be used in laboratories with a radioactive material license. We recently developed novel bioluminescent binding assays for several protein/peptide hormones using the brightest bioluminescent reporter known to date, nanoluciferase (NanoLuc)...
May 2016: Amino Acids
https://www.readbyqxmd.com/read/27018236/detection-of-antibodies-in-blood-plasma-using-bioluminescent-sensor-proteins-and-a-smartphone
#20
Remco Arts, Ilona den Hartog, Stefan E Zijlema, Vito Thijssen, Stan H E van der Beelen, Maarten Merkx
Antibody detection is of fundamental importance in many diagnostic and bioanalytical assays, yet current detection techniques tend to be laborious and/or expensive. We present a new sensor platform (LUMABS) based on bioluminescence resonance energy transfer (BRET) that allows detection of antibodies directly in solution using a smartphone as the sole piece of equipment. LUMABS are single-protein sensors that consist of the blue-light emitting luciferase NanoLuc connected via a semiflexible linker to the green fluorescent acceptor protein mNeonGreen, which are kept close together using helper domains...
April 19, 2016: Analytical Chemistry
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