Imbalance in nucleotide pools of myeloid leukemia cells and HL-60 cells: correlation with cell cycle phase, cell proliferation and differentiation.
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Cell Cycle↗
Biomedical subjects
D de Korte
Publications and source records attributed to D de Korte.
Abnormal nucleotide pattern in the eosinophils of a patient with acute lymphoblastic leukemia associated with eosinophilia.
In this article we present a patient with acute lymphoblastic leukemia (ALL) associated with eosinophilia, in which the eosinophilia preceded a meningeal and bone-marrow relapse of ALL. We analysed the purine and pyrimidine nucleotide content of the eosinophils (92% pure) and compared the nucleotide pattern with that of eosinophils from healthy donors and from patients with eosinophilia not associated with leukemia. The ratios of purine:pyrimidine and of uracil:cytosine nucleotides were decreased compared with those in eosinophils from healthy donors and from patients with eosinophilia with other aetiologies. The total nucleotide concentration was increased, especially the concentration of UDP-sugars and pyrimidine nucleotides. The decrease in these ratios and the increase in concentration of the nucleotides and the UDP-sugars were also detected in leukemic cells of patients with ALL (de Korte et al., Leukemia Res. 10, 389-396 (1986) compared to normal lymphocytes. We suggest a malignant character of the eosinophils in our patient with ALL associated with eosinophilia, in contrast with the non-malignant state suggested previously for these cells.
Adult↗
Anion-exchange high performance liquid chromatography method for the quantitation of nucleotides in human blood cells.
An anion-exchange high performance liquid chromatography (HPLC) method is described for the quantitation of intracellular purine and pyrimidine nucleotides. With an ammonium phosphate salt and pH gradient, complete separation is achieved of all major nucleotides and several interfering substances, such as dehydroascorbic acid and NAD. For optimal resolution of the monophosphates, strict control of the equilibration pH is essential. To prevent interference by a degradation product of NADPH with the determination of GDP, the pH of the high-ionic strength buffer has to be in the range of 4.9-5.0. The use of radially compressed, prepacked cartridges filled with Partisil-10 SAX appeared to be a fast and cheap alternative for expensive stainless-steel columns. The use of ammonium phosphate buffers, in combination with precolumns filled with pellicular silica and SAX resin, and interim EDTA washes prevents baseline shift. This allows analysis at 0.01 Absorbance Units Full Scale during the entire column lifetime (about 180 analyses), which is sufficiently sensitive for the quantitation of low levels of nucleotides, especially when the amount of sample is limited. The usefulness of the presented chromatographic system is demonstrated by the quantitation of the nucleotides in extracts of lymphocytes and neutrophils from the blood of healthy human donors. With this method nucleotide concentrations were measured, with a within-assay variation of 5-10% and an inter-donor variation of 10%.
Blood Cells↗
Nucleotide profiles of normal human blood cells determined by high-performance liquid chromatography.
An anion-exchange high-performance liquid chromatography method has been used to quantitate the intracellular purine and pyrimidine nucleotides in extracts of pure lymphocytes, monocytes, neutrophils, eosinophils, erythrocytes, and platelets isolated from the blood of healthy human donors. For accurate and reproducible measurements of the nucleotide profiles in different types of pure leukocytes, the cell suspensions have to be free of platelets and erythrocytes. Incubation of the purified leukocytes for 1 h at 0 degrees C did not alter the nucleotide concentrations but reduced the interdonor variation to 10%. Incubation of purified lymphocytes for 1 h at 37 degrees C caused considerable changes in the relative concentrations of the adenine, guanine, uracil, and cytosine nucleotides. During this incubation the cell viability, the cell number, and the ATP:ADP ratio decreased. Incubation of monocytes and granulocytes for 1 h at 37 degrees C caused considerable loss of cells and/or cell death. For erythrocytes and platelets reproducible nucleotide concentrations were obtained after extraction of freshly isolated cells. During storage of erythrocytes, both at 0 degrees C and at 37 degrees C, a decrease in the ATP:ADP ratio was detected. In all cell types the predominant nucleotides were purine nucleotides, especially adenosine triphosphate. The relative concentrations of the adenine, guanine, uracil, and cytosine nucleotides were very reproducible per cell type and appeared to be characteristic for each cell type. The total nucleotide content was nearly the same for all cell types except erythrocytes, when expressed per microgram of protein. The described methods for purification and storage of blood cells will be useful for comparison of blood cells from healthy donors with those of patients, for example, leukemia patients, in which deviations of the purine and pyrimidine metabolic enzymes have already been described.
Blood Cells↗
Effect of pre-incubation at 37 degrees C of platelet concentrates on the post-transfusion platelet adhesion capacity to collagen and fibrinogen.
The effect of warming (37 degrees C) of stored platelet concentrates (PC) on the post-transfusion platelet function as measured by the adhesion capacity in a rectangular perfusion system under flow conditions was analyzed in 22 patients undergoing transfusion for stable thrombocytopenia. Nine patients received a PC stored at 22 degrees C and incubated at 37 degrees C for 1 h before transfusion, 10 patients received a non-warmed PC, 3 patients received both a pre-warmed and a non-warmed PC. In the PC the platelet adhesion capacity to collagen was higher in the pre-warmed PC than in the non-warmed PC (33 +/- 5.9% coverage vs. 22 +/- 4.7% coverage, respectively, in a selected group with the same platelet concentration). The adhesion capacity to collagen of the platelets in the patient's blood, measured 10 min after transfusion, had increased considerably in both patient groups and 4 h later the adhesion capacity in both patient groups was similar to that of the pre-warmed PC before transfusion. We conclude that though pre-warming of stored PC had a beneficial effect on the adhesion capacity to collagen of the platelets in the PC, the clinical significance is questionable because already 10 min after transfusion the adhesion capacity to collagen of stored non-warmed platelets had improved to the level of the pre-warmed platelets and 4 h after transfusion this improvement was still present.
Adult↗