In a 2-yr old girl a hemolytic anemia was present since birth requiring multiple blood transfusions. Pyruvate kinase deficiency was suspected on the basis of a marginal enzyme activity, but could not be established due to the presence of massive numbers of donor cells in her peripheral blood. However, by density fractionation we succeeded in the isolation of a small fraction of the patient's own cells, in which a severe pyruvate kinase deficiency could be detected. In contrast hexokinase and glucose-6-phosphate dehydrogenase activities were extremely high, which is indicative that a very immature cell population is present in this fraction. In immunofluorescence studies a clear crossreaction was apparent with anti M2-type pyruvate kinase antibodies, whereas only a faint reaction with anti L-type could be detected. Despite the presence of a slight amount of L-type immunoreactive material, the residual activity in the patient's cell fraction could only be attributed to M2-type pyruvate kinase as was shown by cellulose acetate electrophoresis.

Protein phosphorylation plays an important role in signal transduction of both normal and neoplastic cells. Since increased protein tyrosine phosphorylation may be associated with malignant transformation, we studied the activities of protein tyrosine kinases (PTK) and protein tyrosine phosphatases (PTPase) in patients with various head and neck tumors. Furthermore, we determined the patterns of tyrosine phosphorylated protein (P-tyr) in tissues by western blotting. Enzyme activities were studied in tumor and histologically normal, non-tumorous tissues of 54 patients and in 11 controls. P-tyr patterns were determined in 3 patients and 2 controls. PTK and PTPase activities were greater in tumor tissues than in normal tissue of the cancer patients as well as controls. P-tyr levels in tumors were also higher than in normal tissues. Additionally, PTK activity in normal tissue of tumor patients was significantly higher than in normal tissue of the control group. The same trend was observed for the PTPase activity and P-tyr levels.

Oncogenes play an important role in the process of malignant transformation. Since many of the protein tyrosine kinases (PTK) are products of oncogenes, the aim of this study was to demonstrate whether an increased PTK activity could be found in head and neck tumors. By using a non-radioactive dot-blot assay, PTK activity was measured in tumor and normal tissues of 38 patients with laryngeal cancer. The control group consisted of 19 healthy persons. PTK activity in tumor cells was significantly higher (P < 0.001) than in normal cells of the tumor patients and normal controls. Additionally, the PTK activity in the normal mucosa of the tumor patients was significantly higher than in the normal mucosa of the control group.