4.2.3. Modeling of Therapeutic Hypothermia

Pups were randomly assigned to either normothermic or hypothermic recovery immediately after termination of hypoxia for a duration of 6 h. Normothermic recovery was maintained at a temperature of 37 °C, while hypothermic recovery was maintained at 30 °C according to the previously described protocol [65]. Pups were placed in an open container in a water bath to achieve the desired temperatures. In this experimental set, there were three groups of animals: intact rats (n = 13), rats with HIE and hypothermia (n = 16), and rats with HIE and normothermia (n = 14) (Figure 1C). As in the previous experiments, dry blood spots (DBS) were collected after 6 h of hypoxic exposure for further analysis.