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Rings were mounted on a small vessel wire myograph (Danish Myotech), placed under a resting tension of 1 g and allowed to equilibrate.
Arterial segments (1.0 to 1.5 mm long) were mounted in a small vessel wire myograph (Dual Wire Myograph System 410A, DMT A/S, Aarhus, Denmark).
For short-term exposures, aortic rings were mounted on intra-luminal wires in a small vessel wire myograph [19,20] containing DMEM without l-Arg (37 °C, continuously perfused with 95%O2: 5% CO2) [14].
Tracheal rings containing 3 to 4 cartilage rings were mounted at isometric tension in a small vessel wire myograph (DMT 610 M, DMT-USA, Atlanta, GA) in Krebs bicarbonate buffer containing 119 mM NaCl, 4.7 mM KCl, 1.2 mM MgSO4, 1.2 mM KH2PO4, 2.5 mM CaCl2, 25 mM NaHCO3, 0.03 mM EDTA, and 5.5 mM glucose and continuously gassed with 95% O2: 5% CO2.
The arteries were then cut into segments (1.28 ± 0.04 mm long) and mounted as ring-shaped preparations in the Mulvany-Halpern style small vessel wire myograph chamber (Dual Wire Myograph System 410A, DMT A/S, Aarhus, Denmark) to determine the vascular reactivity during isometric conditions as described elsewhere [ 33].
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When necessary, especially for the smallest vessels, wire incannulation was performed using fluoroscopy.
Planimetric measurements of lesion area correlated well with those made from histological sections subsequently produced from the same vessels (wire-injury: R2 = 0.92; ligation-injury: R2 = 0.89; atherosclerosis: R2 = 0.85), confirming both the accuracy of this methodology and its non-destructive nature.
In the defence cuts of that winter, HMS Endurance, the navy's "trip wire" vessel in the region, was withdrawn, leaving just 50 squaddies at Port Stanley - who duly surrendered when faced with overwhelming odds.
The endothelium was removed by gently rubbing the intimal layer of the vessel with a wire.
It has been demonstrated that vessels in a wire myograph are less sensitive to and respond differently to a variety of agonists (i.e. difference in maximum response and slope of the curve) when compared to vessels in a pressurized system or in vivo [31, 32].
This process, which includes the specification of angioblasts towards venous and arterial fates, along with differential mechanisms of EC migration and subsequent specification within a tissue-specific niche, is highly relevant to our understanding of blood vessel formation and wiring during disease states and tissue regeneration.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com