scholarly journals Detailed imaging of mitochondrial transport and precise manipulation of mitochondrial function with genetically-encoded photosensitizers in adult Drosophila neurons

2021 ◽  
Author(s):  
Francesca Mattedi ◽  
George Chennell ◽  
Alessio Vagnoni
Keyword(s):  
Real Time ◽  
Mitochondrial Function ◽  
Super Resolution ◽  
Neuronal Homeostasis ◽  
Regulation Of Transport ◽  
Detailed Imaging ◽  
Precise Distribution ◽  
Adult Drosophila

Abstract Precise distribution of mitochondria is essential for maintaining neuronal homeostasis. Although detailed mechanisms governing the transport of mitochondria have emerged, it is still poorly understood how the regulation of transport is coordinated in space and time within the physiological context of an organism. How alteration in mitochondrial functionality may trigger changes in organellar dynamics also remains unclear in this context. Therefore, the use of genetically-encoded tools to perturb mitochondrial functionality in real time would be desirable. Here we describe methods to interfere with mitochondrial function with high spatiotemporal precision with the use of photosensitisers in vivo in the intact wing nerve of adult Drosophila. We also provide details on how to visualise the transport of mitochondria and to improve the quality of the imaging to attain super-resolution in this tissue.

Mitochondrial function in vivo evaluated by NADH fluorescence: from animal models to human studies

2007 ◽  
Vol 292 (2) ◽  
pp. C615-C640 ◽  
Author(s):  
Avraham Mayevsky ◽  
Gennady G. Rogatsky
Keyword(s):  
Animal Models ◽  
Real Time ◽  
Mitochondrial Function ◽  
The Body ◽  
Patient Treatment ◽  
Metabolic State ◽  
Experimental Conditions ◽  
Fluorescence Measurements ◽  
Nadh Fluorescence

Normal mitochondrial function is a critical factor in maintaining cellular homeostasis in various organs of the body. Due to the involvement of mitochondrial dysfunction in many pathological states, the real-time in vivo monitoring of the mitochondrial metabolic state is crucially important. This type of monitoring in animal models as well as in patients provides real-time data that can help interpret experimental results or optimize patient treatment. The goals of the present review are the following: 1) to provide an historical overview of NADH fluorescence monitoring and its physiological significance; 2) to present the solid scientific ground underlying NADH fluorescence measurements based on published materials; 3) to provide the reader with basic information on the methodologies used in the past and the current state of the art fluorometers; and 4) to clarify the various factors affecting monitored signals, including artifacts. The large numbers of publications by different groups testify to the valuable information gathered in various experimental conditions. The monitoring of NADH levels in the tissue provides the most important information on the metabolic state of the mitochondria in terms of energy production and intracellular oxygen levels. Although NADH signals are not calibrated in absolute units, their trend monitoring is important for the interpretation of physiological or pathological situations. To understand tissue function better, the multiparametric approach has been developed where NADH serves as the key parameter. The development of new light sources in UV and visible spectra has led to the development of small compact units applicable in clinical conditions for better diagnosis of patients.

SHEDDING LIGHT ON LIFE: OPTICAL ASSESSMENT OF MITOCHONDRIAL FUNCTION AND TISSUE VITALITY IN BIOLOGY AND MEDICINE

2008 ◽  
Vol 01 (01) ◽  
pp. 71-83 ◽  
Author(s):  
AVRAHAM MAYEVSKY
Keyword(s):  
Blood Flow ◽  
Real Time ◽  
Mitochondrial Function ◽  
Foley Catheter ◽  
The Body ◽  
Tissue Blood Flow ◽  
Mitochondrial Activity ◽  
New Device ◽  
Urethral Wall

The involvement of mitochondrial dysfunction in various pathophysiological conditions, developed in experimental and clinical situations, is widely documented. Nevertheless, real time monitoring of mitochondrial function In-vivo is very rare. The pressing question is how the mitochondria of intact tissues behave under In-vivo conditions as compared to isolated mitochondria that had been described by Chance and Williams over 50 years ago. This subject has been recently discussed in detail (Mayevsky and Rogatsky 2007). We reviewed the subject of evaluating mitochondrial function by monitoring NADH fluorescence together with microcirculatory blood flow, Hemoglobin oxygenation and tissue reflectance. These 4 parameters represent the vitality of the tissue and could be monitored in vivo, using optical spectroscopy, in animal models as well as in clinical practice. It is a well known physiological hypothesis that, under emergency conditions, the sympathetic nervous system will give preference to the most vital organs in the body, namely the brain, heart and adrenal glands. The less vital organs, such as the skin, GI-tract, and Urethral wall, will become hypoperfused and their mitochondrial activity will be inhibited. The monitoring of the less vital organs may reveal critical tissue conditions that may manifest an early phase of body deterioration. The aim of the current presentation is to review the experimental and preliminary clinical results accumulated using a new integrated medical device – the "CritiView" which enabled, for the first time, monitoring 4 parameters from the tissue using a single optical probe. The CritiView is a computerized optical device that integrates hardware and software in order to provide real time information on tissue vitality. In preliminary clinical testing, we used a 3-way Foley catheter that includes a bundle of optical fibers enabling the monitoring of the 4 parameters, representing the vitality of the urethral wall (a less vital organ).We found that the exposure of patients to metabolic imbalances in the operation room led to changes in tissue blood flow and inhibition of mitochondrial function in the urethral wall. In conclusion, the new device "CritiView" could provide reliable, real time data on mitochondrial function and tissue vitality in experimental animals as well as in patients.

scholarly journals Study on the Quality Evaluation of Compound Danshen Preparations Based on the xCELLigence Real-Time Cell-Based Assay and Pharmacodynamic Authentication

Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2090 ◽  
Author(s):  
Guojun Yan ◽  
Zhitao Zhu ◽  
Liliang Jin ◽  
Jun Chen ◽  
Hui Xie ◽  
...  
Keyword(s):  
Biological Activity ◽  
Real Time ◽  
Quality Evaluation ◽  
Blood Stasis ◽  
Verification Methods ◽  
Dependent Cell ◽  
Response Profiles ◽  
Dose Dependent

Objective: To perform a preliminary study on the quality evaluation of compound Danshen preparations based on the xCELLigence Real-Time Cell-based Assay (RTCA) system and make a pharmacodynamics verification. Methods: The compound Danshen was discussed as a methodological example, and the bioactivity of the compound Danshen preparations were evaluated by real-time cell electronic analysis technology. Meanwhile, an in vivo experiment on an acute blood stasis rat model was performed in order to verify this novel evaluation through the curative effect of dissipating blood stasis. Results: We determined the cell index (CI) and IC50 of the compound Danshen preparations and produced time/dose-dependent cell response profiles (TCRPs). The quality of the three kinds of compound Danshen preparations was evaluated through the RTCA data. The trend of CI and TCRPs reflected the effect of drugs on the cell (promoting or inhibiting), and it was verified that the results correlated with the biological activity of the drugs using a pharmacodynamics experiment. Conclusion: The RTCA system can be used to evaluate the quality of compound Danshen Preparations, and it can provide a new idea and new method for quantitatively characterizing the biological activity of traditional Chinese medicines (TCMs).

scholarly journals Development of a Quick Quantitative Real-Time PCR for the In Vivo Detection and Quantification of Peach latent mosaic viroid

Plant Disease ◽  
2011 ◽  
Vol 95 (2) ◽  
pp. 137-142 ◽  
Author(s):  
Olivier Parisi ◽  
Philippe Lepoivre ◽  
M. Haissam Jijakli
Keyword(s):  
Real Time ◽  
Plant Pathogens ◽  
Low Cost ◽  
Extraction Procedure ◽  
Reaction Conditions ◽  
Routine Testing ◽  
In Vivo Detection ◽  
Peach Trees

Viroids are plant pathogens infecting a broad range of herbaceous and tree crops. Among them, the Peach latent mosaic viroid (PLMVd) infects mainly peach trees, causing a loss of production with no curative options. Detecting this viroid is thus important for certification procedures aiming to avoid the release of infected material into orchards. Presented here is a complete detection method based on reverse transcription (RT) followed by a quantitative real-time polymerase chain reaction (PCR). New primers were selected and optimal reaction conditions determined for routine application of the method. The technique is 105 times more sensitive than the endpoint RT-PCR used for PLMVd detection, and permits earlier detection of PLMVd in infected plants. The quick, low-cost extraction procedure used and the quality of the results obtained make this method suitable for routine testing.

scholarly journals In Vivo Imaging of the Adult Drosophila melanogaster Heart With Real-Time Optical Coherence Tomography

Circulation ◽  
2006 ◽  
Vol 114 (2) ◽  
Author(s):  
Michael A. Choma ◽  
Susan D. Izatt ◽  
Robert J. Wessells ◽  
Rolf Bodmer ◽  
Joseph A. Izatt
Keyword(s):  
Optical Coherence Tomography ◽  
Drosophila Melanogaster ◽  
Real Time ◽  
In Vivo Imaging ◽  
Optical Coherence ◽  
Adult Drosophila

scholarly journals Signed Real-Time Delay Multiply and Sum Beamforming for Multispectral Photoacoustic Imaging

2018 ◽  
Vol 4 (10) ◽  
pp. 121 ◽  
Author(s):  
Thomas Kirchner ◽  
Franz Sattler ◽  
Janek Gröhl ◽  
Lena Maier-Hein
Keyword(s):  
Open Source ◽  
Real Time ◽  
Photoacoustic Imaging ◽  
Signal To Noise Ratio ◽  
Ultrasound Transducers ◽  
Full Spectrum ◽  
Simple Implementation

Reconstruction of photoacoustic (PA) images acquired with clinical ultrasound transducers is usually performed using the Delay and Sum (DAS) beamforming algorithm. Recently, a variant of DAS, referred to as Delay Multiply and Sum (DMAS) beamforming has been shown to provide increased contrast, signal-to-noise ratio (SNR) and resolution in PA imaging. The main reasons for the use of DAS beamforming in photoacoustics are its simple implementation, real-time capability, and the linearity of the beamformed image to the PA signal. This is crucial for the identification of different chromophores in multispectral PA applications. In contrast, current DMAS implementations are not responsive to the full spectrum of sound frequencies from a photoacoustic source and have not been shown to provide a reconstruction linear to the PA signal. Furthermore, due to its increased computational complexity, DMAS has not been shown yet to work in real-time. Here, we present an open-source real-time variant of the DMAS algorithm, signed DMAS (sDMAS), that ensures linearity in the original PA signal response while providing the increased image quality of DMAS. We show the applicability of sDMAS for multispectral PA applications, in vitro and in vivo. The sDMAS and reference DAS algorithms were integrated in the open-source Medical Imaging Interaction Toolkit (MITK) and are available as real-time capable implementations.

scholarly journals LncRNA UCA1 Promotes Mitochondrial Function of Bladder Cancer via the MiR-195/ARL2 Signaling Pathway

2017 ◽  
Vol 43 (6) ◽  
pp. 2548-2561 ◽  
Author(s):  
Hui-Jin Li ◽  
Xiao-Min Sun ◽  
Zheng-Kun Li ◽  
Qian-Wen Yin ◽  
Huan Pang ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cell Viability ◽  
Real Time ◽  
Signaling Pathway ◽  
Mitochondrial Function ◽  
Real Time Pcr ◽  
Ectopic Expression ◽  
Metabolic Reprogramming ◽  
Cell Counting

Background/Aims: This study aims to identify whether Urothelial Cancer Associated 1 (UCA1) regulates mitochondrial metabolic reprogramming in bladder cancer, and to explore how UCA1 participates in mitochondrial metabolism by the UCA1/miR-195/ARL2 signaling pathway; these findings may be aid in the development of tumor diagnostic and therapeutic strategies. Methods: Bladder tissues were obtained from patients. Stable cell lines were constructed, with ectopic expression of UCA1 in UMUC2 cells and knockdown of UCA1 in 5637 cells. The expression levels of UCA1, miR-195, and ARL2 were detected by real-time PCR, western blotting, and immunohistochemistry Cell viability was detected by Cell Counting Kit-8 (CCK8) assay; mitochondrial DNA copy numbers were tested by realtime PCR; ATP level was evaluated by ATP assay kit; mitochondrial membrane potential was analyzed by 5,5’,6,6’-tetrachloro-1,1’,3,3’- tetraethylbenzimidazolylcarbocyanine iodide (JC-1) fluorescent probe. miRNAs between UCA1 and ARL2 were predicted by TargetScan and RNAHybrid, and then determined by real-time PCR. Dual-luciferase activity assay and RNA immunoprecipitation (RIP) assay were used to verify the relationship between UCA1 and miR-195. The expression level of ARL2 was silenced by small interfering RNA(siRNA). For in vivo experiments, UCA1-silencing 5637 cells were subcutaneously injected into BALB/C nude mice to evaluate the effects of UCA1 on tumor progression by the regulation of miR-195 and ARL2. Results: We demonstrate here that UCA1 enhances mitochondrial function in bladder cancer cells. UCA1 contributes to ARL2-induced mitochondrial activity, which plays an important role in mitochondrial function. UCA1, as a competing endogenous RNA (ceRNA), regulates mitochondrial function through upregulating ARL2. In this way, it inhibited the miR-195 signaling pathway to enhance mitochondrial function in bladder cancer. Additionally, ARL2 is a direct target of miR-195 and can be repressed by either miR-195 overexpression or UCA1 inhibition. Knockdown of ARL2 was analogous to the inhibition of UCA1 and the upregulation of miR-195. Animal experiments further indicated that UCA1 promoted bladder tumor growth by regulating miR-195 /ARL2. Conclusion: These data suggest that UCA1 enhanced mitochondrial function and cell viability through the UCA1/miR-195/ARL2 axis in vitro and in vivo. The elucidation of this signaling network provides a more adequate theoretical basis for understanding the molecular pathology of bladder cancer, and also UCA1 as a potential diagnosis and treatment target for bladder cancer.

Development and Application of Quality Control System for Highway Subgrade Construction

2019 ◽  
Vol 2 (5) ◽  
Author(s):  
Tong Wang
Keyword(s):  
Quality Control ◽  
Real Time ◽  
Detection Method ◽  
System Development ◽  
Effective Control ◽  
Construction Process ◽  
Development Mode ◽  
The Road ◽  
Whole Process

The compaction quality of the subgrade is directly related to the service life of the road. Effective control of the subgrade construction process is the key to ensuring the compaction quality of the subgrade. Therefore, real-time, comprehensive, rapid and accurate prediction of construction compaction quality through informatization detection method is an important guarantee for speeding up construction progress and ensuring subgrade compaction quality. Based on the function of the system, this paper puts forward the principle of system development and the development mode used in system development, and displays the development system in real-time to achieve the whole process control of subgrade construction quality.

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