Evidence map›Paper›PMID 33273578›Full record

ArticleScientific reports2020

NADPH diaphorase detects S-nitrosylated proteins in aldehyde-treated biological tissues.

James M Seckler, Jinshan Shen, Tristan H J Lewis, Mohammed A Abdulameer, Khalequz Zaman, Lisa A Palmer, James N Bates, Michael W Jenkins, Stephen J Lewis

Open access · goldAbstract read
In one paragraph

Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.

0numbers the graph read from it
0cells of the map it votes in
20citing papers in PubMed
1.9field-weighted citation impact, top 13% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

20 citing papers in PubMed, 31 citations in OpenAlex.

  1. Article
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  7. Red light mediates the exocytosis of vasodilatory vesicles from cultured endothelial cells: a cellular, and ex vivo murine model.Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology · 2024
    Article
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  16. Photolytic Measurement of TissueMolecules (Basel, Switzerland) · 2022
    Article
  17. Article
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors at 4 institutions in 1 country.

James M SecklerDepartment of Pediatrics, Case Western Reserve University, Cleveland, OH, 44106, USA.
Jinshan ShenDepartment of Pharmacology, University of Iowa, Iowa City, IA, 52242, USA.
Tristan H J LewisDepartment of Pharmacology and Physiology, University of Georgia, Athens, GA, 30602, USA.
Mohammed A AbdulameerDepartment of Pediatrics, Case Western Reserve University, Cleveland, OH, 44106, USA.
Khalequz ZamanDepartment of Pediatrics, Case Western Reserve University, Cleveland, OH, 44106, USA.
Lisa A PalmerDepartment of Pediatrics, University of Virginia, Charlottesville, VA, 801366, USA.
James N BatesDepartment of Anesthesia, University of Iowa, Iowa City, IA, 52242, USA.
Michael W JenkinsDepartment of Pediatrics, Case Western Reserve University, Cleveland, OH, 44106, USA.
Stephen J LewisDepartment of Pediatrics, Case Western Reserve University, Cleveland, OH, 44106, USA. sjl78@case.edu.
Case Western Reserve University · USUniversity of Iowa · USUniversity of Georgia · USUniversity of Virginia · US

Funding

S-nitrosothiol signaling in pulmonary epithelial cellsP01HL101871 · NHLBI · UNIVERSITY OF VIRGINIA · PI GASTON, BENJAMIN, LEWIS, STEPHEN JOHN · 2011 to 2015
$8.7M
NHLBI NIH HHS P01 HL101871
6 · The paper itself

Abstract

NADPH diaphorase is used as a histochemical marker of nitric oxide synthase (NOS) in aldehyde-treated tissues. It is thought that the catalytic activity of NOS promotes NADPH-dependent reduction of nitro-blue tetrazolium (NBT) to diformazan. However, it has been argued that a proteinaceous factor other than NOS is responsible for producing diformazan in aldehyde-treated tissues. We propose this is a NO-containing factor such as an S-nitrosothiol and/or a dinitrosyl-iron (II) cysteine complex or nitrosated proteins including NOS. We now report that (1) S-nitrosothiols covalently modify both NBT and TNBT, but only change the reduction potential of NBT after modification, (2) addition of S-nitrosothiols or β- or α-NADPH to solutions of NBT did not elicit diformazan, (3) addition of S-nitrosothiols to solutions of NBT plus β- or α-NADPH elicited rapid formation of diformazan in the absence or presence of paraformaldehyde, (4) addition of S-nitrosothiols to solutions of NBT plus β- or α-NADP did not produce diformazan, (5) S-nitrosothiols did not promote NADPH-dependent reduction of tetra-nitro-blue tetrazolium (TNBT) in which all four phenolic rings are nitrated, (6) cytoplasmic vesicles in vascular endothelial cells known to stain for NADPH diaphorase were rich in S-nitrosothiols, and (7) procedures that accelerate decomposition of S-nitrosothiols, markedly reduced NADPH diaphorase staining in tissue sections subsequently subjected to paraformaldehyde fixation. Our results suggest that NADPH diaphorase in aldehyde-fixed tissues is not enzymatic but is due to the presence of NO-containing factors (free SNOs or nitrosated proteins such as NOS), which promote NADPH-dependent reduction of NBT to diformazan.

Indexed as

AnimalsAzo CompoundsBrain StemCerebellumFormaldehydeMaleNADPH DehydrogenaseNitric Oxide SynthaseNitroblue TetrazoliumOxidation-ReductionPolymersRatsRats, Sprague-DawleyS-NitrosothiolsStaining and LabelingAzo Compoundsdiformazan dyeFormaldehydeNADPH DehydrogenaseNitric Oxide SynthaseNitroblue TetrazoliumparaformPolymersS-Nitrosothiols

Identifiers

PMID33273578
PMCPMC7713249
OpenAlexW3109888177

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.