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X Demographics
Mendeley readers
Attention Score in Context
| Title |
A Novel Stopped-Flow Assay for Quantitating Carbonic-Anhydrase Activity and Assessing Red-Blood-Cell Hemolysis
|
|---|---|
| Published in |
Frontiers in Physiology, March 2017
|
| DOI | 10.3389/fphys.2017.00169 |
| Pubmed ID | |
| Authors |
Pan Zhao, R. Ryan Geyer, Walter F. Boron |
| Abstract |
We report a novel carbonic-anhydrase (CA) assay and its use for quantitating red-blood-cell (RBC) lysis during stopped-flow (SF) experiments. We combine two saline solutions, one containing HEPES/pH 7.03 and the other, ~1% CO2/44 mM [Formula: see text]/pH 8.41, to generate an out-of-equilibrium CO2/[Formula: see text] solution containing ~0.5% CO2/22 [Formula: see text]/pH ~7.25 (10°C) in the SF reaction cell. CA catalyzes relaxation of extracellular pH to ~7.50: [Formula: see text] + H(+) → CO2 + H2O. Proof-of-concept studies (no intact RBCs) show that the pH-relaxation rate constant (kΔpH)-measured via pyranine fluorescence-rises linearly with increases in [bovine CAII] or [murine-RBC lysate]. The y-intercept (no CA) was kΔpH = 0.0183 s(-1). Combining increasing amounts of murine-RBC lysate with ostensibly intact RBCs (pre-SF hemolysis ≅0.4%)-fixing total [hemoglobin] at 2.5 μM in the reaction cell to simulate hemolysis from ostensibly 0 to 100%-causes kΔpH to increase linearly. This y-intercept (0% lysate/100% ostensibly intact RBCs) was kΔpH = 0.0820 s(-1), and the maximal kΔpH (100% lysate/0% intact RBCs) was 1.304 s(-1). Thus, mean percent hemolysis in the reaction cell was ~4.9%. Phenol-red absorbance assays yield indistinguishable results. The increase from 0.4 to 4.9% presumably reflects mechanical RBC disruption during rapid mixing. In all fluorescence studies, the CA blocker acetazolamide reduces kΔpH to near-uncatalyzed values, implying that all CA activity is extracellular. Our lysis assay is simple, sensitive, and precise, and will be valuable for correcting for effects of lysis in physiological SF experiments. The underlying CA assay, applied to blood plasma, tissue-culture media, and organ perfusates could assess lysis in a variety of applications. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Switzerland | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 16 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 16 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 25% |
| Researcher | 4 | 25% |
| Professor | 1 | 6% |
| Student > Master | 1 | 6% |
| Student > Bachelor | 1 | 6% |
| Other | 0 | 0% |
| Unknown | 5 | 31% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 2 | 13% |
| Agricultural and Biological Sciences | 2 | 13% |
| Engineering | 2 | 13% |
| Immunology and Microbiology | 1 | 6% |
| Medicine and Dentistry | 1 | 6% |
| Other | 1 | 6% |
| Unknown | 7 | 44% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 28 March 2017.
All research outputs
#20,411,380
of 22,961,203 outputs
Outputs from Frontiers in Physiology
#9,439
of 13,712 outputs
Outputs of similar age
#268,927
of 308,511 outputs
Outputs of similar age from Frontiers in Physiology
#167
of 215 outputs
Altmetric has tracked 22,961,203 research outputs across all sources so far. This one is in the 1st percentile – i.e., 1% of other outputs scored the same or lower than it.
So far Altmetric has tracked 13,712 research outputs from this source. They typically receive more attention than average, with a mean Attention Score of 7.6. This one is in the 1st percentile – i.e., 1% of its peers scored the same or lower than it.
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We're also able to compare this research output to 215 others from the same source and published within six weeks on either side of this one. This one is in the 1st percentile – i.e., 1% of its contemporaries scored the same or lower than it.