title: Repeatability of the evaluation of systemic microvascular endothelial function using laser doppler perfusion monitoring: clinical and statistical implications.
abstract: OBJECTIVE
  An awareness of the repeatability of biological measures is required to properly design and calculate sample sizes for longitudinal interventional studies. We investigated the day-to-day repeatability of measures of systemic microvascular reactivity using laser Doppler perfusion monitoring.
  
  
  METHODS
  We performed laser Doppler perfusion monitoring in combination with skin iontophoresis using acetylcholine and sodium nitroprusside as well as post-occlusive reactive and thermal hyperemia twice within two weeks. The repeatability was assessed by calculating the within-subject standard deviations, limits of agreement, typical errors and intra-class correlation coefficients between days 1 and 2. The ratio of the within-subject standard deviation to the mean values obtained on days 1 and 2 (within-subject standard deviation/GM) was used to determine the condition with the best repeatability.
  
  
  RESULTS
  Twenty-four healthy subjects, aged 24.6 ± 3.8 years, were recruited. The area under the curve of the vasodilatory response to post-occlusive reactivity showed marked variability (within-subject standard deviation/GM = 0.83), while the area under the curve for acetylcholine exhibited less variability (within-subject standard deviation/ GM = 0.52) and was comparable to the responses to sodium nitroprusside and thermal treatment (within-subject standard deviations/GM of 0.67 and 0.56, respectively). The area under the blood flow/time curve for vasodilation during acetylcholine administration required the smallest sample sizes, the area under the blood flow/time curve during post-occlusive reactivity required the largest sample sizes, and the area under the blood flow/time curves of vasodilation induced by sodium nitroprusside and thermal treatment required intermediate sizes.
  
  
  CONCLUSIONS
  In view of the importance of random error related to the day-to-day repeatability of laser Doppler perfusion monitoring, we propose an original and robust statistical methodology for use in designing prospective clinical studies.
authors.affiliation: Diabetes Unit, State University of Rio de Janeiro, Brazil. etibi@ioc.fiocruz.br
authors.forename: Eduardo
authors.initials: E
authors.lastname: Tibiriçá
authors.forename: Alessandra S M
authors.initials: AS
authors.lastname: Matheus
authors.forename: Bruno
authors.initials: B
authors.lastname: Nunes
authors.forename: Sandro
authors.initials: S
authors.lastname: Sperandei
authors.forename: Marilia B
authors.initials: MB
authors.lastname: Gomes
mesh_terms: D000109:Acetylcholine; D000328:Adult; D003430:Cross-Sectional Studies; D004730:Endothelium, Vascular / Q000502:physiology*; D005260:Female; D006801:Humans; D007478:Iontophoresis / Q000379:methods*; D017078:Laser-Doppler Flowmetry / Q000379:methods*; D008137:Longitudinal Studies; D008297:Male; D008833:Microcirculation / Q000502:physiology*; D008991:Monitoring, Physiologic / Q000379:methods; D009599:Nitroprusside; D015203:Reproducibility of Results; D012867:Skin / Q000098:blood supply*; D014664:Vasodilation; D014665:Vasodilator Agents
chemical_list: D014665:Vasodilator Agents; D009599:Nitroprusside; D000109:Acetylcholine