Gait analysis was performed on 15 male rats (10 weeks old) to evaluate their walking pattern. To obtain footprints, four paws were colored with different nontoxic water colors and animal was trained to walk through an ally (width, 8 cm; length, 120 cm; height, 10 cm), leading to its home cage. After training, animals were divided randomly into three groups: (i) direct exposure, (ii) poly-Oxime gel–treated, and (iii) naïve control. Footprints were analyzed manually, and SFI was calculated using following formula (40, 41)Embedded Imagewhere N indicates normal/before exposure, E indicates after exposure, TOF is the distance to the opposite foot, PL is the distance from the heel to the third toe (the print length), TS is the distance from the first toe to the fifth toe (the toe spread), and IT is the distance from the second toe to the fourth toe (the intermediate toe spread).

EMG of animals from these three groups was recorded using Muscle SpikerBox (Backyard Brains). Skin surface electrodes were used with adhesive pads and conductive gel to facilitate the recordings. To track the muscle spasms under anesthesia (2.5% isoflurane in carbogen), EMG from biceps femoris of left hind limb was recorded. To avoid animal from disturbing the electrodes, we recorded EMG between spinotrapezius and gluteus maximus when the animal was awake.

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