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The supernatant was discarded and 100 l DMSO was added and mixed gently for 10 min while avoiding light

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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

The obesity trial studied a population averaging about 240 lb at BMI 37, while lean-recomposition reports describe people well under 180 lb