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Office arterial blood pressure was assessed with a UA-779 automatic digital device (A&D Instruments LTD., Abingdon, Oxon, UK), taking one measurement on each arm after a minimum of 5 min at rest, seated upright.
The plates were shaken vigorously at room temperature for 10 min and incubated for another 10 min at rest.
After 10 min at rest, the absorbance of the solutions of the tube was read at 490 nm.
The instantaneous HR was acquired and monitored by an HR monitor (Polar® S810i) during 10 min at rest pre-exercise, 4 min of DBT, 5 min of exercise protocol (active hand and foot exercises, and active-assisted lower extremities exercises) and 10 min at rest post-exercise in the supine position.
The record was collected in two situations: 10 min at rest in the sitting position during spontaneous breathing; and 10 min at the same position with BiPAP (BiPAP®; Tranquility Respironics) application through a nasal mask.
The plates were incubated at 42°C and shaken intermittently at 900 r.p.m. (87 sec shaking, 33 min at rest) using a FLUOstar OMEGA microplate reader (BMG Labtech) in a double orbital configuration.
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Subjects received visual stimulation for 2 min at resting baseline and recovery and every 5 min after the onset of exercise until exhaustion.
For tumours analysis, blocks containing breast tumours of 4 µm thick sections were cut, deparaffinized and subjected to antigen retrieval by water bath in citrate buffer (pH 6.0) during 40 min, and at rest during 20 min. Then, the slides were treated according to the manufacturer's instructions (Duolink II Fluorescence, Olink Bioscience, Sweden).
Fig. 2A shows DAF-FM fluorescence from adult and old mice over 60 min, both at rest and where fibers were stimulated to contract for two 10 min periods (indicated by black bars).
The respiratory rate was expressed as breaths per min (BPM) at rest.
A second identical stimulation period was then applied followed by a 10 min period at rest [31].
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com