How reproducible is bilateral forearm plethysmography?

Abstract

Aims: In studies using strain-gauge forearm plethysmography to measure changes in forearm blood flow (FBF) during intra-arterial infusions of vasoactive substances, measurements are often made in both arms simultaneously and the change in ratio of the infused and control arms used to express responses. However, the reproducibility of bilateral plethysmography in this setting has not been addressed in published studies. The unilateral technique remains in use, and forearm vascular resistance (FVR), an alternative method of expressing responses, is used by some investigators. We have assessed: (a) the intra-subject variability of bilateral FBF measurements (FBF ratios) at rest, after unilateral forearm exercise, and during intra-arterial infusions of vasoconstrictor substances; (b) whether bilateral plethysmography is more reproducible than unilateral plethysmography; and (c) the reproducibility of FVR (unilateral and bilateral).

Methods: Study 1 Nine healthy subjects attended 3 study days, 1 week apart. FBF was measured at rest and after 2 min of standardized unilateral forearm exercise; between-day intra-subject variability was expressed as coefficients of variation (CV) calculated using two-way analysis of variance (ANOVA). Study 2 Five healthy subjects attended 2 study days when FBF was measured during incremental infusions of noradrenaline (15, 30, 150, 300 pmol min[-1]) and angiotensin II (1, 5, 10, 50 pmol min[-1]); for each individual subject at each dose intra-subject variability was assessed using the difference between responses (percentage change from baseline) on days 1 and 2.

Results: Study 1 At rest, intra-subject variability (CV) of baseline FBF ratios was 19% compared with 31% (left) and 39% (right) for unilateral FBF measurements. After ipsilateral exercise, unilateral FBF measurements were more reproducible (32 vs 17%) than FBF ratios; by 20 min after exercise, the previous pattern had been re-established (19 vs 27%). Intra-subject variability (CV) of baseline FVR ratio and post-exercise FVR was 14%. Study 2 Inter-quartile ranges of the differences between responses on days 1 and 2 (FBF ratios vs FBF) were: angiotensin II 14 vs 18%; noradrenaline 16 vs 27%.

Conclusions: FBF ratios are more reproducible than unilateral FBF measurements at rest (CV 19% vs 39%) and for measuring responses to intra-arterial infusions of vasoconstrictor substances. FVR may have a small reproducibility advantage. Non-experimental stimuli can cause significant and misleading changes in measured responses if unilateral measurements are used; it is therefore recommended that responses to intra-arterial infusions should be measured using bilateral forearm plethysmography with the results expressed as FBF ratios.

Figure 1

Experimental protocols for Study 1 (upper panel) and Study 2 (lower panel). Bold bars represent 3 min periods of FBF measurements (see Methods). B1 = baseline 1, B2 = baseline 2.

Figure 2

Individual FBF (left and right) and FBF ratios in nine healthy male subjects on days 1, 2 and 3. Data are shown for Baseline 1 (left panel), Unilateral exercise (middle panel), and 20 min post-exercise (right panel).

Figure 3

Grouped (mean±s.e. mean, % change from baseline) FBF (left and right) and FBF ratios in five healthy male subjects during intra-arterial infusions of angiotensin II (left) and noradrenaline (right) on day 1 (▪) and day 2 (□).

Figure 4

Individual FBF (left and right) and FBF ratios (% change from baseline) in five healthy male subjects during intra-arterial infusions of angiotensin II on day 1 (closed symbols) and day 2 (open symbols).

Figure 5

Individual FBF (left and right) and FBF ratios (% change from baseline) in five healthy male subjects during intra-arterial infusions of noradrenaline on day 1 (closed symbols) and day 2 (open symbols).

Figure 6

Difference between responses on days 1 and 2 plotted against mean response for each individual subject at each dose for both FBF and FBF ratios (see Methods).