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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 Biol+Proced+Online
2018 ; 20
(ä): 7
Nephropedia Template TP
gab.com Text
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English Wikipedia
DNA Area and NETosis Analysis (DANA): a High-Throughput Method to Quantify
Neutrophil Extracellular Traps in Fluorescent Microscope Images
#MMPMID29618953
Rebernick R
; Fahmy L
; Glover C
; Bawadekar M
; Shim D
; Holmes CL
; Rademacher N
; Potluri H
; Bartels CM
; Shelef MA
Biol Proced Online
2018[]; 20
(ä): 7
PMID29618953
show ga
BACKGROUND: Neutrophil extracellular traps (NETs), extracellular structures
composed of decondensed chromatin and antimicrobial molecules, are released in a
process called NETosis. NETs, which are part of normal host defense, have also
been implicated in multiple human diseases. Unfortunately, methods for
quantifying NETs have limitations which constrain the study of NETs in disease.
Establishing optimal methods for NET quantification holds the potential to
further elucidate the role of NETs in normal and pathologic processes. RESULTS:
To better quantify NETs and NET-like structures, we created DNA Area and NETosis
Analysis (DANA), a novel ImageJ/Java based program which provides a simple,
semi-automated approach to quantify NET-like structures and DNA area. DANA can
analyze many fluorescent microscope images at once and provides data on a per
cell, per image, and per sample basis. Using fluorescent microscope images of
Sytox-stained human neutrophils, DANA quantified a similar frequency of NET-like
structures to the frequency determined by two different individuals counting by
eye, and in a fraction of the time. As expected, DANA also detected increased DNA
area and frequency of NET-like structures in neutrophils from subjects with
rheumatoid arthritis as compared to control subjects. Using images of
DAPI-stained murine neutrophils, DANA (installed by an individual with no
programming background) gave similar frequencies of NET-like structures as the
frequency of NETs determined by two individuals counting by eye. Further, DANA
quantified more NETs in stimulated murine neutrophils compared to unstimulated,
as expected. CONCLUSIONS: DANA provides a means to quantify DNA decondensation
and the frequency of NET-like structures using a variety of different fluorescent
markers in a rapid, reliable, simple, high-throughput, and cost-effective manner
making it optimal to assess NETosis in a variety of conditions.