PMID- 29945605
OWN - NLM
STAT- MEDLINE
DCOM- 20181011
LR  - 20181114
IS  - 1746-6148 (Electronic)
IS  - 1746-6148 (Linking)
VI  - 14
IP  - 1
DP  - 2018 Jun 27
TI  - A novel approach to identifying and quantifying neutrophil extracellular trap 
      formation in septic dogs using immunofluorescence microscopy.
PG  - 210
LID - 10.1186/s12917-018-1523-z [doi]
LID - 210
AB  - BACKGROUND: Canine neutrophils release neutrophil extracellular traps (NETs) in 
      response to lipopolysaccharide but NETs from clinical septic dogs had not been 
      identified. The primary aim is to describe the methodology of identifying and 
      quantifying neutrophil extracellular traps (NETs) in cytology samples of septic 
      foci in dogs with sepsis using immunofluorescence microscopy. Cytology 
      samples including endotracheal tracheal wash (ETW), bronchoalveolar lavage (BAL), 
      abdominal and pleural effusion collected from 5 dogs (3 septic, 2 non-septic) 
      were fixed, permeabilized and stained for myeloperoxidase (MPO), citrullinated 
      histone H3 (citH3) and cell-free DNA (cfDNA). Fluorescence microscopy was used to 
      identify and quantify NETs in 10 random views at 40x magnification. NETs were 
      identified based on co-localization of MPO, citH3 and cfDNA. NETs were quantified 
      as a ratio (number of NETs: number of neutrophils). Neutrophils were identified 
      based on cytoplasmic MPO, cellular diameter and nuclear morphology. RESULTS: NETs 
      were identified and quantified in all cytology samples collected from septic 
      dogs. A small number of NETs was documented in one dog with sterile chronic 
      bronchitis. No NETs were found in sterile abdominal effusion collected from one 
      dog with congestive heart failure. CONCLUSIONS: Immunofluorescence microscopy 
      could be a useful tool for the study of NETs in dogs with clinical sepsis.
FAU - Li, Ronald H L
AU  - Li RHL
AUID- ORCID: 0000-0002-3325-2744
AD  - Department of Veterinary Surgical and Radiological Sciences, School of Veterinary 
      Medicine, University of California, One Shield Avenue, Davis, California, 95161, 
      USA. rhli@ucdavis.edu.
FAU - Johnson, Lynelle R
AU  - Johnson LR
AD  - Department of Medicine and Epidemiology, School of Veterinary Medicine, 
      University of California, Davis, California, USA.
FAU - Kohen, Casey
AU  - Kohen C
AD  - William Pritchard Veterinary Medical Teach Hospital, University of California, 
      Davis, California, USA.
FAU - Tablin, Fern
AU  - Tablin F
AD  - Department of Anatomy, Physiology and Cell Biology, School of Veterinary 
      Medicine, University of California, Davis, California, USA.
LA  - eng
GR  - D15CA-907/Morris Animal Foundation/
GR  - 2016-24-F/School of Veterinary Medicine, University of California, Davis/
PT  - Journal Article
DEP - 20180627
PL  - England
TA  - BMC Vet Res
JT  - BMC veterinary research
JID - 101249759
SB  - IM
MH  - Animals
MH  - Bronchoalveolar Lavage Fluid/cytology
MH  - Dog Diseases/*diagnosis
MH  - Dogs
MH  - Extracellular Traps/*metabolism
MH  - Microscopy, Fluorescence/methods/*veterinary
MH  - Sepsis/diagnosis/*veterinary
PMC - PMC6020318
OTO - NOTNLM
OT  - Cell-free DNA
OT  - Citrullinated histones
OT  - Peptidylarginine deiminase
OT  - Sepsis
COIS- ETHICS APPROVAL: The study protocol was approved by the Institutional Animal Care 
      and Use Committee at the University of California, Davis. All samples were 
      collected after informed written consent was obtained from the owners. CONSENT 
      FOR PUBLICATION: Not applicable. COMPETING INTERESTS: The authors declare that 
      they have no competing interests.
EDAT- 2018/06/28 06:00
MHDA- 2018/10/12 06:00
PMCR- 2018/06/27
CRDT- 2018/06/28 06:00
PHST- 2017/11/20 00:00 [received]
PHST- 2018/06/12 00:00 [accepted]
PHST- 2018/06/28 06:00 [entrez]
PHST- 2018/06/28 06:00 [pubmed]
PHST- 2018/10/12 06:00 [medline]
PHST- 2018/06/27 00:00 [pmc-release]
AID - 10.1186/s12917-018-1523-z [pii]
AID - 1523 [pii]
AID - 10.1186/s12917-018-1523-z [doi]
PST - epublish
SO  - BMC Vet Res. 2018 Jun 27;14(1):210. doi: 10.1186/s12917-018-1523-z.