## 4 Discussion
Our findings suggest that, despite reductions in protein levels, liver function as assessed by ALT and ALP may not have been significantly impacted by either alcohol or benzodiazep exposure in this model. A challenge when investigating the globulin concentration in hypothermia is also represented by certain conditions that can affect protein values, such as trauma. Traumain combination with hypothermia can cause co-agulopathies that can affect the globulin concentration. Traumatic injuries, especially those involving significant blood loss, environmental exposure, or severe pain, can precipitate hypothermia by disrupting thermoregulation and increasing metabolic demands. Trauma triggers an inflammatory response, which can lead to the increased production of globulins, particularly immuno-globulins. This response is aimed at combating potential infections and facilitating healing. In cases of significant blood loss due to trauma, the concentration of globulins can be affected by hemodilution if fluid resuscitation is employed. This may lead to falsely low measurements of globulin levels despite ongoing inflammatory responses. In traumatic cases, the combination of injury and hypothermia can significantly impact globulin levels. Elevated globulins may indicate an inflammatory response to trauma. Such conditions make it difficult to determine whether it is primary hypothermia that has resulted in a change in the globulin concentration or secondary hypothermia after trauma, which also affects the globulin concentration [35]. In our study, the protein levels varied significantly among the groups (\(p<0.001\) for total protein and \(p=0.002\) for globulin, respectively). Post hoc tests confirmed these differences for the comparisons of alcohol vs. control and benzodiazepine vs. control comparisons, but not between the alcohol and benzodiazepine groups. This indicates the potential of globulin to be used as a parameter for distinguishing primary from secondary hypothermia. The amylase levels were significantly different among the groups (\(p<0.001\)), with post hoc tests confirming significant differences between all pairs: alcohol vs. benzodiazepine (\(p=0.002\)), alcohol vs. control (\(p=0.003\)), and benzodiazepine vs. control (\(p<0.001\)). Elevated amylase levels can indicate pancreatic stress or damage. In the context of hypothermia, elevated amylase might be associated with secondary complications like pancreatitis or stress from severe cold exposure [36; 37]. Hypothermia can reduce the metabolic rate and slow down enzymatic reactions. As temperatures drop, the activity of amylase may decrease, potentially leading to lower serum amylase concentrations. In conditions leading to secondary hypothermia, such as pancreatitis or sepsis, elevated amylase levels may occur due to tissue damage or inflammation. In these cases, hypothermia may coexist with elevated amylase, complicating the clinical picture; for example, acute pancreatitis occur as an underlying disease or be caused by some chemical agent [38]. The context in which hypothermia occurs is crucial, and while this study did not investigate the possible interaction of alcohol and benzodiazepines, this is worth exploring in future research using animal models, as CNS depressants are shown to have multiplicative effects on the thermoregulatory response when taken in combination [39].