scholarly journals Quantitative imaging of intracellular nanoparticle exposure enables prediction of nanotherapeutic efficacy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Qingqing Yin ◽  
Anni Pan ◽  
Binlong Chen ◽  
Zenghui Wang ◽  
Mingmei Tang ◽  
...  
Keyword(s):  
Quantitative Imaging ◽  
Tumour Mass ◽  
Nanoparticle Exposure ◽  
Imaging Tool ◽  
Intracellular Compartments ◽  
Signal Output ◽  
Extracellular Microenvironment ◽  
Intracellular Targets ◽  
Ph Variations ◽  
Therapeutic Responses

AbstractNanoparticle internalisation is crucial for the precise delivery of drug/genes to its intracellular targets. Conventional quantification strategies can provide the overall profiling of nanoparticle biodistribution, but fail to unambiguously differentiate the intracellularly bioavailable particles from those in tumour intravascular and extracellular microenvironment. Herein, we develop a binary ratiometric nanoreporter (BiRN) that can specifically convert subtle pH variations involved in the endocytic events into digitised signal output, enabling the accurately quantifying of cellular internalisation without introducing extracellular contributions. Using BiRN technology, we find only 10.7–28.2% of accumulated nanoparticles are internalised into intracellular compartments with high heterogeneity within and between different tumour types. We demonstrate the therapeutic responses of nanomedicines are successfully predicted based on intracellular nanoparticle exposure rather than the overall accumulation in tumour mass. This nonlinear optical nanotechnology offers a valuable imaging tool to evaluate the tumour targeting of new nanomedicines and stratify patients for personalised cancer therapy.

Quantitative imaging of translocated silver following nanoparticle exposure by laser ablation-inductively coupled plasma-mass spectrometry

2018 ◽  
Vol 10 (8) ◽  
pp. 836-840 ◽  
Author(s):  
David P. Bishop ◽  
Mandy Grossgarten ◽  
Dörthe Dietrich ◽  
Antje Vennemann ◽  
Nerida Cole ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Ablation ◽  
Inductively Coupled Plasma ◽  
Quantitative Imaging ◽  
Nanoparticle Exposure ◽  
Inductively Coupled ◽  
Icp Ms ◽  
Plasma Mass Spectrometry

We demonstrate the use of LA-ICP-MS for determining the location and quantification of silver in a rat spleen following nanoparticle exposure.

scholarly journals Gravin-associated kinase signaling networks coordinate γ-tubulin organization at mitotic spindle poles

2020 ◽  
Vol 295 (40) ◽  
pp. 13784-13797
Author(s):  
Paula J. Bucko ◽  
Irvin Garcia ◽  
Ridhima Manocha ◽  
Akansha Bhat ◽  
Linda Wordeman ◽  
...  
Keyword(s):  
Quantitative Imaging ◽  
Subcellular Location ◽  
Asymmetric Distribution ◽  
Binding Partner ◽  
Mitotic Kinases ◽  
Spindle Poles ◽  
Biochemical Assays ◽  
Anchoring Protein ◽  
Intracellular Compartments ◽  
Genetic Deletion

Mitogenic signals that regulate cell division often proceed through multienzyme assemblies within defined intracellular compartments. The anchoring protein Gravin restricts the action of mitotic kinases and cell-cycle effectors to defined mitotic structures. In this report we discover that genetic deletion of Gravin disrupts proper accumulation and asymmetric distribution of γ-tubulin during mitosis. We utilize a new precision pharmacology tool, Local Kinase Inhibition, to inhibit the Gravin binding partner polo-like kinase 1 at spindle poles. Using a combination of gene-editing approaches, quantitative imaging, and biochemical assays, we provide evidence that disruption of local polo-like kinase 1 signaling underlies the γ-tubulin distribution defects observed with Gravin loss. Our study uncovers a new role for Gravin in coordinating γ-tubulin recruitment during mitosis and illuminates the mechanism by which signaling enzymes regulate this process at a distinct subcellular location.

Quantitative Imaging Of the Green Fluorescent Proteins

1998 ◽  
Vol 4 (S2) ◽  
pp. 1004-1005
Author(s):  
David W. Piston ◽  
George H. Patterson ◽  
Susan M. Knobel
Keyword(s):  
Fluorescent Proteins ◽  
Protein Localization ◽  
Quantitative Imaging ◽  
Cloning And Expression ◽  
Green Fluorescent Proteins ◽  
Specific Gene ◽  
Full Potential ◽  
Imaging Tool ◽  
Naturally Occurring ◽  
Green Fluorescent

The cloning and expression of GFP in heterologous systems introduced a fantastic tool for studying specific gene expression and protein localization inside living cells. However, one aspect of GFP that has not been exploited to its full potential is its use as a quantitative imaging tool. To determine its quantitative usefulness, we have addressed five points that are important in GFP imaging: detectable signal over background, photostability, pH stability of the molecule, temperature dependence of chromophore formation, and estimation and normalization of GFP levels.To determine the quantitative limits of GFP in cells, several GFP versions (wtGFP, αGFP (F99S/M153T/V163A), S65T, EGFP (F64L/S65T), and a blue-shifted variant, EBFP (F64L/S65T/Y66H/Y145F)) were compared by imaging of GFP expressing cells or by spectroscopic measurements of purified proteins. When imaged, the GFP signals are contaminated by the naturally occurring background autofluorescence, but improved detection can be achieved for each green GFP by combination of confocal microscopy using 488 nm excitation, a rapid cut-on dichroic mirror, and a narrow bandpass emission filter (Figure l).

scholarly journals Imaging and Electrophysiology for Degenerative Cervical Myelopathy [AO Spine RECODE DCM Research Priority Number 9]

2021 ◽  
pp. 219256822110574
Author(s):  
Allan R. Martin ◽  
Lindsay Tetreault ◽  
Benjamin M. Davies ◽  
Armin Curt ◽  
Patrick Freund ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Cervical Myelopathy ◽  
Tissue Injury ◽  
Quantitative Imaging ◽  
Cord Compression ◽  
Narrative Review ◽  
Imaging Tool ◽  
Spinal Cord Dysfunction ◽  
Degenerative Cervical Myelopathy ◽  
And Function

Study Design Narrative review. Objective The current review aimed to describe the role of existing techniques and emerging methods of imaging and electrophysiology for the management of degenerative cervical myelopathy (DCM), a common and often progressive condition that causes spinal cord dysfunction and significant morbidity globally. Methods A narrative review was conducted to summarize the existing literature and highlight future directions. Results Anatomical magnetic resonance imaging (MRI) is well established in the literature as the key imaging tool to identify spinal cord compression, disc herniation/bulging, and inbuckling of the ligamentum flavum, thus facilitating surgical planning, while radiographs and computed tomography (CT) provide complimentary information. Electrophysiology techniques are primarily used to rule out competing diagnoses. However, signal change and measures of cord compression on conventional MRI have limited utility to characterize the degree of tissue injury, which may be helpful for diagnosis, prognostication, and repeated assessments to identify deterioration. Early translational studies of quantitative imaging and electrophysiology techniques show potential of these methods to more accurately reflect changes in spinal cord microstructure and function. Conclusion Currently, clinical management of DCM relies heavily on anatomical MRI, with additional contributions from radiographs, CT, and electrophysiology. Novel quantitative assessments of microstructure, perfusion, and function have the potential to transform clinical practice, but require robust validation, automation, and standardization prior to uptake.

scholarly journals Gravin-associated kinase signaling networks coordinate γ-tubulin organization at mitotic spindle poles

2020 ◽  
Author(s):  
Paula J. Bucko ◽  
Irvin Garcia ◽  
Ridhima Manocha ◽  
Akansha Bhat ◽  
Linda Wordeman ◽  
...  
Keyword(s):  
Quantitative Imaging ◽  
Subcellular Location ◽  
Asymmetric Distribution ◽  
Binding Partner ◽  
Mitotic Kinases ◽  
Spindle Poles ◽  
Biochemical Assays ◽  
Anchoring Protein ◽  
Intracellular Compartments ◽  
Genetic Deletion

AbstractMitogenic signals that regulate cell division often proceed through multi-enzyme assemblies within defined intracellular compartments. The anchoring protein Gravin restricts the action of mitotic kinases and cell-cycle effectors to defined mitotic structures. In this report we discover that genetic deletion of Gravin disrupts proper accumulation and asymmetric distribution of γ-tubulin during mitosis. We utilize a new precision pharmacology tool, Local Kinase Inhibition (LoKI), to inhibit the Gravin binding partner polo-like kinase 1 (Plk1) at spindle poles. Using a combination of gene-editing approaches, quantitative imaging, and biochemical assays we provide evidence that disruption of local Plk1 signaling underlies the γ-tubulin distribution defects observed with Gravin loss. Our study uncovers a new role for Gravin in coordinating γ-tubulin recruitment during mitosis and illuminates the mechanism by which signaling enzymes regulate this process at a distinct subcellular location.

scholarly journals Correction: Quantitative imaging of translocated silver following nanoparticle exposure by laser ablation-inductively coupled plasma-mass spectrometry

2018 ◽  
Vol 10 (8) ◽  
pp. 926-926 ◽  
Author(s):  
David P. Bishop ◽  
Mandy Grossgarten ◽  
Dörthe Dietrich ◽  
Antje Vennemann ◽  
Nerida Cole ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Ablation ◽  
Inductively Coupled Plasma ◽  
Quantitative Imaging ◽  
Nanoparticle Exposure ◽  
Inductively Coupled ◽  
Plasma Mass Spectrometry

Correction for ‘Quantitative imaging of translocated silver following nanoparticle exposure by laser ablation-inductively coupled plasma-mass spectrometry’ by David P. Bishop et al., Anal. Methods, 2018, DOI: 10.1039/c7ay02294h.

scholarly journals Intracellular delivery of therapeutic antibodies into specific cells using antibody-peptide fusions

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Julie Gaston ◽  
Nicolas Maestrali ◽  
Guilhem Lalle ◽  
Marie Gagnaire ◽  
Alessandro Masiero ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Cell Penetrating Peptides ◽  
Therapeutic Modality ◽  
Therapeutic Antibodies ◽  
Cell Penetration ◽  
C Terminus ◽  
Macromolecular Drugs ◽  
Cell Penetrating ◽  
Intracellular Compartments ◽  
Intracellular Targets

AbstractBecause of their favorable properties as macromolecular drugs, antibodies are a very successful therapeutic modality for interfering with disease-relevant targets in the extracellular space or at the cell membrane. However, a large number of diseases involve cytosolic targets and designing antibodies able to efficiently reach intracellular compartments would expand the antibody-tractable conditions. Here, we genetically fused cell penetrating peptides (CPPs) at various positions to an antibody targeting cancer cells, evaluated the developability features of the resulting antibody-peptide fusions and the ability of selected constructs to reach the cytosol. We first determined positions in the IgG structure that were permissive to CPP incorporation without destabilizing the antibody. Fusing CPPs to the C-terminus of the light chain and either before or after the hinge had the least effect on antibody developability features. These constructs were further evaluated for cell penetration efficiency. Two out of five tested CPPs significantly enhanced antibody penetration into the cytosol, in particular when fused before or after the hinge. Finally, we demonstrate that specific antibody binding to the cell surface target is necessary for efficient cell penetration of the CPP-antibody fusions. This study provides a solid basis for further exploration of therapeutic antibodies for intracellular targets.

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