Ozonation of Human Blood Induces a Remarkable Upregulation of Heme Oxygenase-1 and Heat Stress Protein-70

Heme oxygenase-I (HO-1) has emerged as one of the most protective enzymes and its pleiotropic activities have been demonstrated in a variety of human pathologies. Unpublished observations have shown that HO-1 is induced after the infusion of ozonated blood into the respective donors, and many other experimental observations have demonstrated the efficacy of oxidizing agents. It appeared worthwhile to evaluate whether we could better define the activity of potential inducers such as hydrogen peroxide and ozonated human plasma. Human vascular endothelial cells at confluence were challenged with different concentrations of these inducers and the simultaneous production of nitric oxide (NO); and HO-1 was measured by either measuring nitrite, or bilirubin formation, or/and the immune reactivity of the protein by Western blot using a rabbit antihuman HO-1 and Hsp-70. The results show that production of both NO and HO-1 is fairly dose dependent but is particularly elevated using human plasma after transient exposure to a medium ozone concentration. At this concentration, there is also induction of Hsp-70. The results clarify another positive effect achievable by the use of ozone therapy.

Polypeptide Pattern of Apple Tissues Affected by Calcium-related Physiopathologies

Polypeptides from the apple pulp of Smoothee Golden Delicious and White Renete apples were resolved by 1-D denaturing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). According to the electropherograms, there were lower concentrations of 88, 74, 70.6 and 47.5-42kDa proteins in bitter pit spots. Proteins weighing 30 and 26kDa were rare in sound pulp but frequently appeared in pits and adjacent tissue. Finally, a novel 18kDa protein was found in bitter pit spots in both varieties, and also in chemically induced corky lesions either by magnesium infiltration or ammonium oxalate cortical injections. The available data suggested that the novel protein might be an inhibitor of pectinmethylesterase, a small heat stress protein (smHSP) or a product of the Ypr-10 gene family identified with ‘Mal d 1’, the main allergen of apples. To elucidate the possible smHSP nature of the 18kDa, a set of apples were heated at 40°C for 20h, developing this protein in both the oxidised tissue and in the adjacent.