PMID: 18214303
Owners commonly notice one or more of the following: Limping or lameness: favoring a limb, "toe-touching," or intermittent limping that comes and goes Stiffness after rest: difficulty rising after sleeping, or a slow, stiff first few minutes that loosens with movement Reluctance to jump, climb, or use stairs: hesitating at the car, couch, or staircase they used to handle easily Reduced activity or stamina: shorter walks, less play, or lagging behind on familiar routes Audible joint changes: clicking or popping from a joint, particularly the stifle (knee) Muscle loss: visible thinning of the muscle on one hind limb compared with the other, a sign of chronic underuse Behavioral changes: licking a joint, irritability when a limb is handled, or a drop in overall mood tied to discomfort A sudden non-weight-bearing lameness the dog will not put any weight on a leg warrants prompt veterinary attention rather than watchful waiting, as it can indicate an acute cruciate rupture or fracture
adding 50mL of deprotection reagent to remove Fmoc for 30 minutes at the temperature of not more than 20 , adding 50mL of DMF to wash for multiple times, and obtaining positive indene detection result
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