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Figure 4: (a) Anxiety related behavioral analysis (I) in open field test (OFT) for Cd2+ and nicotine toxicity. Bar chart representing the center square duration (CSD) in OFT for the treated groups and control after the acute (D1–D7) and chronic (D7–D21) treatment phases. The duration spent in the center square of the open field area was estimated to determine the mean duration for each group at the end of the acute and chronic phases. After the acute treatment phase, no significant change in anxiety linked behavior was seen in the nicotine and (or) cadmium treatment groups. After chronic treatment, cadmium and nicotine treatment increased anxiety linked behavior in the animals, as shown by a significant decrease in CSD (D7–D21). Similarly, a combined nicotine-cadmium treatment also caused an increase in anxiety linked behavior (reduced CSD) when compared with the control (). (b) Anxiety related behavioral analysis (II) in open field test (OFT) for the treatment and control groups. The mean frequency of rearing was determined for the nicotine and (or) treatment groups versus the control after the acute (D1–D7) and chronic (D7–D21) treatment phases. No significant change in the frequency of rearing for the treatment groups after the acute treatment phase. After the chronic phase, subsequent tests show a significant reduction in rearing time (anxiety) for the cadmium treatment group when compared with the control, nicotine, and nicotine-cadmium treatment groups . (c) Anxiety related behavioral analysis (III) in open field test (OFT) for Cd2+ and (or) nicotine toxicity. Bar chart representing the frequency of grooming in OFT for the treatment groups and control after the acute (D1–D7) and chronic (D7–D21) treatment phases. No significant change in the frequency of grooming was observed after the acute treatment phase. Similarly, no significant change was seen in the frequency of grooming after the chronic treatment phase for all treatment groups versus the control.