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Each mouse was manually restrained, and corneal stimulation (0.2 ms pulse, 6 Hz, 3 s) was performed at a current intensity of 17 mA (ECT Unit 57800, Ugo Basile, Comerio, Italy).
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The fat was injected subcutaneously into the scalp using a 14G needle while the animals were manually restrained.
During blood collection, gilts were manually restrained using a pig snare.
Llamas were manually restrained with a halter and lead rope for blood collection.
Captured bats were manually restrained and anesthetized by intravenous injection; ≈0.2 1.0 mL of blood was collected from the propatagial vein before the bat was released.
Fish were manually restrained for <3 min during which time fork length (FL) was measured, tissues were biopsied and a radio transmitter was inserted.
Cattle and goats were manually restrained and had blood samples taken from the jugular vein using 10 ML plain vacutainer tubes.
For experimentally infected pigs and the pigs in cohorts that were repeatedly sampled, the animals were manually restrained and blood was obtained from the anterior vena cava using a vacutainer and 20-gauge needle.
Three kennel-housed coyotes (Canis latrans) were manually restrained and 1 ml of blood was collected via the cephalic vein using a sterile 23 gauge syringe and transferred to sterile 5 ml tubes.
Vaccination methods were the same as described above for Study 1. SL vaccination of unanesthetized animals was performed in a manner similar to that described for anesthetized animals except that animals were manually restrained for at least 1 minute after administration.
The resulting model was manually completed in Coot [35] and restrained-structure-refinement implementing TLS refinement [36] resulted in cAMP model with Rwork and Rfree of 20.6% and 23.0% respectively.
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CEO of Professional Science Editing for Scientists @ prosciediting.com