Performance status was recorded using the Palliative Performance Scale (PPS) [19]. All assessments were conducted between 0900 h and 1300 h. All participants were asked to refrain from smoking and caffeine ingestion on the morning of assessment, but were not asked to stop any of their usual medications or fast. A physician and research nurse performed the tests of autonomic function in a quiet room at ambient

temperature (21-23 C). Autonomic function tests were carried out using a modified Ewing’s battery [17]. Heart rate was measured by ECG Inhibitors,research,lifescience,medical using standard limb leads; heart rate (HR) tests were excluded if invalidated by arrhythmia, excessive ectopic activity or movement artefact. Blood pressure (BP) was monitored using the

Finometer Pro device (Finapres Medical Systems BV, Amsterdam, the Netherlands) which enables noninvasive Inhibitors,research,lifescience,medical beat-to-beat BP measurement from finger arterial BP. The BP recordings are derived from the circumferential pressure generated by a finger cuff, which is varied to maintain a constant digital arterial size, as measured Inhibitors,research,lifescience,medical by a photoplethysmograph. Under such conditions the external cuff pressure equals the internal digital arterial pressure [20]. Participants rested in the supine position for at least ten minutes before testing. During this time they were covered with a blanket and wore a thermal mitten with glove liner in order to improve BP signal pick-up. Blood pressure tests were excluded if the trace was obscured by movement artefact or artefact due to external pressure on the finger-cuff. Parasympathetic tests 1. Deep breathing Whilst supine, participants were requested to ‘take slow Inhibitors,research,lifescience,medical deep breaths, so that each breath in lasts five seconds and each breath out lasts 5 s, for a total of six consecutive breaths’. This was rehearsed prior to testing and the Onalespib in vitro tester guided the timing of the breaths for the participant Inhibitors,research,lifescience,medical by verbally counting through each of the six breaths/cycles. The maximum and minimum HR during each breathing cycle was calculated from the corresponding shortest and longest R-R interval, and the response recorded as the mean of the differences during

three successive breathing cycles. 2. Active stand Participants were requested to stand up from the supine position as quickly as possible ADAMTS5 and to remain standing, in silence, for three minutes, with the monitored arm resting by their side. Assistance with rising was provided when this could not be achieved independently. Heart rate response was measured as the ratio of the maximum R-R interval at or around the 30th beat after starting to stand, to the minimum R-R interval at or around the 15th beat. 3. Valsalva manoeuvre The valsalva manoeuvre was achieved by forced expiration, against an open glottis. Participants were requested to achieve a constant pressure of 40 mmHg for 15 s. The procedure was rehearsed prior to testing and the tester guided the participant by counting aloud through the fifteen seconds.