Influence of adrenergic agonists on the release of amino acids from rat skeletal muscle studied by microdialysis

Rosdahl,; Samuelsson,; Ungerstedt,; Henriksson,

doi:10.1046/j.1365-201x.1998.00341.x

Acta Physiol Scand

1998

DOI: 10.1046/j.1365-201x.1998.00341.x

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Influence of adrenergic agonists on the release of amino acids from rat skeletal muscle studied by microdialysis

Rosdahl Rosdahl

Samuelsson Samuelsson

Ungerstedt Ungerstedt

et al.

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“…Thus, the addition of any stabilizing proteins should be avoided if MS is used for detection. Dextrans, with a molecular mass of 60−70 kDa, are other additives that have been used successfully to increase the osmotic pressure. ,,,− Dextran is the collective name for neutral polysaccharides with large dimensions that are used extensively in many biological applications. Dextrans do not affect the MS signal, since they can easily be removed prior to separation and detection.…”

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confidence: 99%

Methodological Aspects on Microdialysis Protein Sampling and Quantification in Biological Fluids: An In Vitro Study on Human Ventricular CSF

et al. 2010

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There is growing interest in sampling of protein biomarkers from the interstitial compartment of the brain and other organs using high molecular cutoff membrane microdialysis (MD) catheters. However, recent data suggest that protein sampling across such MD membranes is a highly complex process that needs to be further studied. Here, we report three major improvements for microdialysis sampling of proteins in complex biological matrixes. The improvements in this in vitro study using human ventricular cerebrospinal fluid as the sample matrix include increased fluid recovery control, decreased protein adsorption on the microdialysis membrane and materials, and novel quantitative mass spectrometry analysis. Dextrans in different concentrations and sizes were added to the perfusion fluid. It was found that dextrans with molecular mass 250 and 500 kDa provided a fluid recovery close to 100%. An improved fluid recovery precision could be obtained by self-assembly triblock polymer surface modification of the MD catheters. The modified catheters also delivered a significantly increased extraction efficiency for some of the investigated proteins. The final improvement was to analyze the dialysates with isobaric tagged (iTRAQ) proteomics, followed by tandem mass spectrometric analysis. By using this technique, 48 proteins could be quantified and analyzed with respect to their extraction efficiencies. The novel aspects of microdialysis protein sampling, detection, and quantification in biological fluids presented in this study should be considered as a first step toward better understanding and handling of the challenges associated with microdialysis sampling of proteins. The next step is to optimize the developed methodology in vivo.

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Methodological Aspects on Microdialysis Protein Sampling and Quantification in Biological Fluids: An In Vitro Study on Human Ventricular CSF

et al. 2010

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“…To overcome these fluid losses, the osmotic pressure difference between the outside sample space and the dialysate has to be compensated for by adding either dextran-70 or additional protein, such as BSA. 10,18 In addition to the problems with fluid loss, the most common analytical method for detecting cytokines is enzyme-linked immunosorbent assay (ELISA). 19,20 The limitation of a standard ELISA method is that only a single cytokine can be analyzed per well.…”

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Enhanced Microdialysis Relative Recovery of Inflammatory Cytokines Using Antibody-Coated Microspheres Analyzed by Flow Cytometry

Sellati

Stenken

2004

Anal. Chem.

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Achieving high relative recovery (RR) of proteins during microdialysis sampling is difficult due to the diffusion limitations inherent to this sampling process. This often causes low microdialysis RR for proteins with molecular weight >10 kDa. A RR enhancement process for microdialysis sampling of proteins has been developed that can be readily used with flow cytometers. Multiplexed RR enhancement and detection of five different cytokines (TNF-alpha, IFN-gamma, IL-2, IL-4, and IL-5) was achieved by including antibody-coated microspheres in the microdialysis perfusion fluid. Inclusion of these antibody-coated microspheres causes an increase in the analyte diffusive driving force across the dialysis membrane and a subsequent increase in the relative recovery. For the five cytokines, typical control and enhanced relative recoveries at a 1.0 microL/min flow rate were as follows (n = 3): TNF-alpha, 5.5 +/- 0.6% and 60.4 +/- 5.8%; IFN-gamma, 2.6 +/- 0.3% and 25.8 +/- 2.3%; IL-5, 1.4 +/- 0.2% and 4.9 +/- 0.1%; IL-4, 10.9 +/- 0.6% and 78.8 +/- 8.0%; and IL-2, 4.1 +/- 0.4% and 19.8 +/- 2.5%. Using this approach, a four- to 12-fold enhancement of microdialysis RR was achieved for the five cytokines from a quiescent solution. The enhancement varies among the five cytokines and may be due to different diffusive and antibody binding properties. TNF-alpha exhibited the highest RR enhancement, while IL-5 exhibited the lowest. Experimental parameters that affect the enhancement, such as flow rate, sample collection volume, and bead density, were studied.

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Influence of adrenergic agonists on the release of amino acids from rat skeletal muscle studied by microdialysis

Cited by 2 publications

References 0 publications

Methodological Aspects on Microdialysis Protein Sampling and Quantification in Biological Fluids: An In Vitro Study on Human Ventricular CSF

Methodological Aspects on Microdialysis Protein Sampling and Quantification in Biological Fluids: An In Vitro Study on Human Ventricular CSF

Enhanced Microdialysis Relative Recovery of Inflammatory Cytokines Using Antibody-Coated Microspheres Analyzed by Flow Cytometry

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