title: Intraobserver and interobserver variability and spatial differences in histologic examination of carotid endarterectomy specimens.
abstract: BACKGROUND
  Studies using histologic examination and protein analysis of atherosclerotic plaques are increasingly being performed, but reproducibility of plaque histology and variation of plaque composition among different parts of the plaque, which are key to reliability of these studies, are relatively unexplored. Therefore, this study investigated the intraobserver and interobserver variability of plaque histology and spatial variability in plaque composition.
  
  
  METHODS
  Atherosclerotic plaques (n = 100) obtained during carotid endarterectomy were divided into 0.5-cm segments. Paraffin sections were stained and semiquantitatively analyzed (four categories: no, minor, moderate, and heavy) for fat, macrophages, smooth muscle cells, collagen, calcification, thrombus, and overall phenotype. First, to determine the intraobserver and interobserver reproducibility, two independent observers independently analyzed the plaques. Second, to investigate spatial variability in plaque composition, histologic appearances of the culprit lesions (0-segment) were compared with the histologic appearances of adjacent (+5 mm) and more distant (+10 mm) plaque segments of 30 specimens.
  
  
  RESULTS
  The kappa values for intraobserver variability of fat, macrophages, smooth muscle cells, collagen, calcifications, thrombus, and overall phenotype were 0.83, 0.85, 0.71, 0.63, 0.81, 0.80, and 0.86, respectively, and kappa values for interobserver variability were 0.68, 0.74, 0.54, 0.59, 0.82, 0.75, and 0.71, respectively. Comparison of the histologic scorings of adjacent segments revealed a mean kappa of 0.40 (range, 0.33 to 0.60). When the culprit segment was compared with the more distant segment, the mean kappa was 0.24; however, in 91% of cases, the difference between the culprit segment and the distal segment was one category or less.
  
  
  CONCLUSIONS
  Semiquantitative analysis of carotid atherosclerotic plaque histology was well reproducible, both intraobserver and interobserver. Although variation between different plaque segments in histologic appearance was observed, differences were small in almost all cases. Variability in histologic examination needs to be taken into account in studies comparing plaque imaging with histopathology and plaque research studies.
authors.affiliation: Department of Vascular Surgery, University Medical Center Utrecht, Utrecht, The Netherlands. w.e.hellings@umcutrecht.nl
authors.forename: Willem E
authors.initials: WE
authors.lastname: Hellings
authors.forename: Gerard
authors.initials: G
authors.lastname: Pasterkamp
authors.forename: Anne
authors.initials: A
authors.lastname: Vollebregt
authors.forename: Cees A
authors.initials: CA
authors.lastname: Seldenrijk
authors.forename: Jean-Paul P M
authors.initials: JP
authors.lastname: De Vries
authors.forename: Evelyn
authors.initials: E
authors.lastname: Velema
authors.forename: Dominique P V
authors.initials: DP
authors.lastname: De Kleijn
authors.forename: Frans L
authors.initials: FL
authors.lastname: Moll
mesh_terms: D000273:Adipose Tissue / Q000473:pathology; D002114:Calcinosis / Q000473:pathology; D002340:Carotid Artery Diseases / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D003094:Collagen / Q000032:analysis; D016894:Endarterectomy, Carotid; D006801:Humans; D008264:Macrophages / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000473:pathology; D009426:Netherlands; D015588:Observer Variation; D010641:Phenotype; D015203:Reproducibility of Results; D012720:Severity of Illness Index; D013927:Thrombosis / Q000473:pathology
chemical_list: D003094:Collagen