The Experts below are selected from a list of 48399 Experts worldwide ranked by ideXlab platform
Yi Yang - One of the best experts on this subject based on the ideXlab platform.
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In Vivo monitorIng of cellular energy metabolism usIng sonar a highly responsive sensor for nad nadh redox state
Nature Protocols, 2016Co-Authors: Yuzheng Zhao, Aoxue Wang, Yejun Zou, Joseph Loscalzo, Yi YangAbstract:NADH and its oxidized form NAD(+) have a central role In energy metabolism, and their concentrations are often considered to be among the most important readouts of metabolic state. Here, we present a detailed protocol to image and monitor NAD(+)/NADH redox state In livIng cells and In Vivo usIng a highly responsive, genetically encoded fluorescent sensor known as SoNar (sensor of NAD(H) redox). The chimeric SoNar proteIn was Initially developed by InsertIng circularly permuted yellow fluorescent proteIn (cpYFP) Into the NADH-bIndIng domaIn of Rex proteIn from Thermus aquaticus (T-Rex). It functions by bIndIng to either NAD(+) or NADH, thus InducIng proteIn conformational changes that affect its fluorescent properties. We first describe steps for how to establish SoNar-expressIng cells, and then discuss how to use the system to quantify the Intracellular redox state. This approach is sensitive, accurate, simple and able to report subtle perturbations of various pathways of energy metabolism In real time. We also detail the application of SoNar to high-throughput chemical screenIng of candidate compounds targetIng cell metabolism In a microplate-reader-based assay, along with In Vivo fluorescence imagIng of tumor xenografts expressIng SoNar In mice. Typically, the approximate time frame for fluorescence imagIng of SoNar is 30 mIn for livIng cells and 60 mIn for livIng mice. For high-throughput chemical screenIng In a 384-well-plate assay, the whole procedure generally takes no longer than 60 mIn to assess the effects of 380 compounds on cell metabolism.
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In Vivo monitorIng of cellular energy metabolism usIng sonar a highly responsive sensor for nad nadh redox state
Nature Protocols, 2016Co-Authors: Yuzheng Zhao, Aoxue Wang, Joseph Loscalzo, Ni Su, Yi YangAbstract:SoNar is a fluorescent biosensor that is able to monitor NAD+/NADH redox state In livIng cells and In Vivo. This protocol describes how to use SoNar for sIngle cell imagIng, high-throughput chemical screenIng, and In Vivo imagIng In mice.
Tsutomu Kawasaki - One of the best experts on this subject based on the ideXlab platform.
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In Vivo monitorIng of plant small gtpase activation usIng a forster resonance energy transfer biosensor
Plant Methods, 2018Co-Authors: Hann Ling Wong, Akira Akamatsu, Masayuki Higuchi, Tomonori Matsuda, Kenichi Kosami, Noriko Inada, Jun Okuda, Qiong Wang, Tsutomu KawasakiAbstract:Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patternIng and hormone responses. Thus, to monitor their activation status withIn plant cells is believed to be the key step In understandIng their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and InteractIng proteIn chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 durIng various defence responses In cells. Here, we describe the protocol for visualizIng spatiotemporal activity of plant Rac/ROP GTPase In livIng plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be adaptable for monitorIng activation for other plant small GTPases and proteIn–proteIn Interactions for other FRET sensors In various plant cells.
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In Vivo monitorIng of plant small gtpase activation usIng a forster resonance energy transfer biosensor
bioRxiv, 2018Co-Authors: Hann Ling Wong, Akira Akamatsu, Masayuki Higuchi, Tomonori Matsuda, Kenichi Kosami, Noriko Inada, Tsutomu Kawasaki, Jun Okuda, Qiong Wang, Shingo NagawaAbstract:Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patternIng and hormone responses. Thus, to monitor their activation status withIn plant cells is believed to be the key step In understandIng their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and InteractIng proteIn chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 durIng various defence responses In rice cells. Here, we describe the protocol for visualizIng spatiotemporal activity of plant Rac/ROP GTPase In livIng plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be widely adaptable for monitorIng activation for other plant small GTPases and for other FRET sensors In various plant cells.
Kartik M Varadarajan - One of the best experts on this subject based on the ideXlab platform.
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Interfacial load monitorIng and failure detection In total joInt replacements via piezoresistive bone cement and electrical impedance tomography
Smart Materials and Structures, 2020Co-Authors: Hamid Ghaednia, Crystal E Owens, Ricardo Roberts, Tyler N Tallman, Anastasios John Hart, Kartik M VaradarajanAbstract:Aseptic loosenIng, or loss of implant fixation, is a common complication followIng total joInt replacement. Revision surgeries cost the healthcare system over $8 billion annually In the United States. Despite the prevalence of aseptic loosenIng, timely and accurate detection remaIns a challenge because traditional imagIng modalities, such as plaIn radiographs, struggle to reliably detect the early stages of implant loosenIng. Motivated by this challenge, we present a novel approach for In Vivo monitorIng and failure detection of cemented joInt replacements. Poly(methyl methacrylate) (PMMA) bone cement is modified with low volume fractions of chopped carbon fiber (CF) to impart piezoresistive-based self-sensIng. Electrical impedance tomography (EIT) is then used to detect and monitor load-Induced deformation and fracture of CF/PMMA In a phantom tank. We therefore show that EIT Indeed is able to detect loadIng force on a prosthetic surrogate, distInguish between IncreasIng load magnitudes, detect failure of implant fixation, and even distInguish between cement crackIng and cement de-bondIng without direct contact to the surrogate. Because EIT is a low-cost, physiologically benign, and potentially real-time imagIng modality, the feasibility study hereIn presented could positively impact orthopedic researchers by providIng, via In Vivo monitorIng, Insight Into the factors that Initiate aseptic loosenIng.
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Interfacial load monitorIng and failure detection In total joInt replacements via piezoresistive bone cement and electrical impedance tomography
arXiv: Signal Processing, 2019Co-Authors: Hamid Ghaednia, Crystal E Owens, Ricardo Roberts, Tyler N Tallman, John A Hart, Kartik M VaradarajanAbstract:Aseptic loosenIng, or loss of implant fixation, is a common complication followIng total joInt replacement. Revision surgeries cost the healthcare system over $8 billion annually In the US. Despite the prevalence of aseptic loosenIng, timely and accurate detection remaIns a challenge because traditional imagIng modalities such as plaIn radiographs struggle to reliably detect the early stages of implant loosenIng. Motivated by this challenge, we present a novel approach for In Vivo monitorIng and failure detection of cemented joInt replacements. Poly(methyl methacrylate) (PMMA) bone cement is modified with low volume fractions of chopped carbon fiber (CF) to impart piezoresistive-based self-sensIng. Electrical impedance tomography (EIT) is then used to detect and monitor load-Induced deformation and fracture of CF/PMMA In a phantom tank. We therefore show that EIT Indeed is able to adeptly detect loadIng force on a prosthetic surrogate, distInguish between IncreasIng load magnitudes, detect failure of implant fixation, and even distInguish between cement crackIng and cement de-bondIng. Because EIT is a low-cost, physiologically benign, and potentially real-time imagIng modality, the feasibility study hereIn presented has potential to transform the standard of care for post-operative monitorIng of implant conditions. Beyond clInical relevance, this technique could positively impact orthopedic researchers by providIng, via In Vivo monitorIng, Insight Into the factors that Initiate aseptic loosenIng.
Akira Akamatsu - One of the best experts on this subject based on the ideXlab platform.
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In Vivo monitorIng of plant small gtpase activation usIng a forster resonance energy transfer biosensor
Plant Methods, 2018Co-Authors: Hann Ling Wong, Akira Akamatsu, Masayuki Higuchi, Tomonori Matsuda, Kenichi Kosami, Noriko Inada, Jun Okuda, Qiong Wang, Tsutomu KawasakiAbstract:Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patternIng and hormone responses. Thus, to monitor their activation status withIn plant cells is believed to be the key step In understandIng their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and InteractIng proteIn chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 durIng various defence responses In cells. Here, we describe the protocol for visualizIng spatiotemporal activity of plant Rac/ROP GTPase In livIng plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be adaptable for monitorIng activation for other plant small GTPases and proteIn–proteIn Interactions for other FRET sensors In various plant cells.
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In Vivo monitorIng of plant small gtpase activation usIng a forster resonance energy transfer biosensor
bioRxiv, 2018Co-Authors: Hann Ling Wong, Akira Akamatsu, Masayuki Higuchi, Tomonori Matsuda, Kenichi Kosami, Noriko Inada, Tsutomu Kawasaki, Jun Okuda, Qiong Wang, Shingo NagawaAbstract:Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patternIng and hormone responses. Thus, to monitor their activation status withIn plant cells is believed to be the key step In understandIng their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and InteractIng proteIn chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 durIng various defence responses In rice cells. Here, we describe the protocol for visualizIng spatiotemporal activity of plant Rac/ROP GTPase In livIng plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be widely adaptable for monitorIng activation for other plant small GTPases and for other FRET sensors In various plant cells.
Hann Ling Wong - One of the best experts on this subject based on the ideXlab platform.
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In Vivo monitorIng of plant small gtpase activation usIng a forster resonance energy transfer biosensor
Plant Methods, 2018Co-Authors: Hann Ling Wong, Akira Akamatsu, Masayuki Higuchi, Tomonori Matsuda, Kenichi Kosami, Noriko Inada, Jun Okuda, Qiong Wang, Tsutomu KawasakiAbstract:Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patternIng and hormone responses. Thus, to monitor their activation status withIn plant cells is believed to be the key step In understandIng their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and InteractIng proteIn chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 durIng various defence responses In cells. Here, we describe the protocol for visualizIng spatiotemporal activity of plant Rac/ROP GTPase In livIng plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be adaptable for monitorIng activation for other plant small GTPases and proteIn–proteIn Interactions for other FRET sensors In various plant cells.
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In Vivo monitorIng of plant small gtpase activation usIng a forster resonance energy transfer biosensor
bioRxiv, 2018Co-Authors: Hann Ling Wong, Akira Akamatsu, Masayuki Higuchi, Tomonori Matsuda, Kenichi Kosami, Noriko Inada, Tsutomu Kawasaki, Jun Okuda, Qiong Wang, Shingo NagawaAbstract:Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patternIng and hormone responses. Thus, to monitor their activation status withIn plant cells is believed to be the key step In understandIng their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and InteractIng proteIn chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 durIng various defence responses In rice cells. Here, we describe the protocol for visualizIng spatiotemporal activity of plant Rac/ROP GTPase In livIng plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be widely adaptable for monitorIng activation for other plant small GTPases and for other FRET sensors In various plant cells.
