31P MRS analysis of the phospholipid composition of the peripheral blood mononuclear cells (PBMC) and bone marrow mononuclear cells (BMMC) of patients with acute leukemia (AL).

The aim of this study was to evaluate the phospholipid concentration in acute leukemia (AL) blast cells from peripheral blood (PBMC) and bone marrow (BMMC). In vitro 31P Nuclear Magnetic Resonance Spectroscopy (31P MRS) was used. The integral intensities of the resonant peaks and the phospholipid concentrations in PBMC and BMMC were analyzed. Differences in the phospholipid concentrations in cells from myeloblastic or lymphoblastic lines were also evaluated. This investigation was carried out on phospholipid extracts from PBMC and BMMC from 15 healthy volunteers and 77 patients with AL (samples taken at the moment of diagnosis). A significant decrease in sphingomyelin (SM) and phosphtidylserine (PS) was observed in the PBMC of patients with AL relative to the results for the healthy volunteers. For ALL, we found a significant decrease in the concentration of phosphatidylcholine plasmalogen (CPLAS), SM, PI+PE (phosphatidylinositol + phosphatidylethanolamine) and PS in comparison with the results for healthy volunteers and patients with AML. Experiments with BMMC cells revealed a significant decrease in the concentration of CPLAS, SM, PI+PE, and PS in ALL relative to AML. Additionally, a significant decrease in phosphatidylcholine (PC) concentration was observed in ALL compared to AML. If the phospholipid extracts were taken simultaneously from the same patient, there were no significant differences in the integral intensities and phospholipid concentrations between PBMC and BMMC.

Platelet-activating factor changes in phospholipid extracts from the plasma, peripheral blood mononuclear cells and bone marrow mononuclear cells of patients with acute leukemia--a 31P MRS in vitro study.

The aim of this investigation was to evaluate the changes in PAF concentrations in the plasma, PBMC and BMMC of patients with acute lymphoblastic leukemia (ALL) and acute myeloblastic leukemia (AML). The plasma was from 23 healthy volunteers (HV) and 44 patients with AL (16 ALL, 28 AML). The PBMC were from 15 HV and 55 patients with AL (18 ALL, 37 AML), and the BMMC from 40 patients with AL (11 ALL, 29 AML). Methanol-chloroform phospholipid extraction from 60 x 10(6) cells (PBMC or BMMC) was performed according to a modified version of Folch's method. (31)P MRS data was obtained on an AMX 300 Bruker spectrometer (7.05 T). The PAF concentration in the plasma of the patients with ALL or AML was lower than that for the healthy volunteers. The PAF concentration in the plasma of the patients with ALL did not differ significantly from that of the patients with AML. In the case of both the PBMC and BMMC, the PAF concentration was significantly diminished in patients with ALL relative to the concentration for those with AML and for the healthy volunteers. No differences were observed in the PAF concentrations for the AML patients and the healthy volunteers.

Application of 31P MRS to the analysis of phospholipid changes in plasma of patients with acute leukemia.

The aim of the experiment was to evaluate the changes of phospholipid concentrations in patients (n=30) with acute leukemia compared with reference group of healthy volunteers (n=21). The analysis focused on the following phospholipids (PL) collected from plasma: phosphatidylcholine (PC), plasmalogen of phosphatidylcholine (CPLAS), lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylethanolamine (PE), and phosphatidylinositol (PI). Phospholipid extracts were obtained by Folch's method from 4 ml of plasma. 31P MR spectra were obtained on an AMX Bruker 300 MHz (7.05 T) spectrometer. Calculation of concentration based on integral intensity of the phospholipid relative to an internal concentration standard of MDPA. For healthy volunteers, the following values of phospholipid concentrations were obtained: (5.18+/-1.615) mmol/l for PC+CPLAS; (0.364+/-0.178) mmol/l for LPC; (1.211+/-0.411) mmol/l for SM; (0.343+/-0.124) mmol/l for PI+PE. PLs of patients were assayed at least twice: at the time of diagnosis and, when appropriate, at the time of complete remission from the disease (CR). At the time of diagnosis, the mean concentrations of studied compounds were: (1.602+/-0.716) mmol/l for PC+CPLAS; (0.041+/-0.048) mmol/l for LPC; (0.398+/-0.198) mmol/l for SM; (0.045+/-0.071) mmol/l for PI+PE. After attainment of complete remission (CR), the respective values were as follows: (4.094+/-1.886) mmol/l for PC+CPLAS; (0.295+/-0.139) mmol/l for LPC; (1.123+/-0.634) mmol/l for SM; (0.230+/-0.125) mmol/l for PI+PE. All concentrations found in patients at the time of diagnosis were significantly lower than in reference group and in those benefited from complete remission (CR). By contrast the differences in concentrations of phospholipids in plasma between patients with complete remission (CR) and healthy volunteers were no statistically significant.

31P MRS analysis of the phospholipid composition of normal human peripheral blood mononuclear cells (PBMC).

The aim of this investigation was to characterize the phospholipid composition of normal human blood mononuclear cells using 31P NMR spectroscopy. Mononuclear cells of peripheral blood were obtained from 10 volunteers. Phospholipid extracts were prepared from 60x10(6) cells according to modified Folch's method. An AMX 300 Bruker spectrometer 7.05 T was used. The 31P spectrum of phospholipid extracts from normal human PBMC consisted of 9 peaks, with one each for phosphatidylcholine (PC), plasmalogen of phosphatidylcholine (CPLAS), lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS) and cardiolipin (CL), and another one due to the external reference substance, methylenediphosphonic acid (MDPA). The concentrations of these phospholipids (PL), based on the integral intensities, were as follows: 0.398 +/- 0.078 mmole/l for PC; 0.033 +/- 0.019 mmole/l for CPLAS; 0.155 +/- 0.043 mmole/l for SM; 0.266 +/- 0.104 mmole/l for PI+PE; 0.101 +/- 0.040 mmole/l for PS, and 0.026 +/- 0.033 mmole/l for CL. The results of this study confirmed that 31P MRS is a convenient tool for measuring the phospholipid concentrations of biological samples.

Bone marrow transplantation in the course of hematological malignancies--follow-up study in blood serum by 31P MRS.

BACKGROUND: Bone marrow transplantation (BMT) is one of the most efficient methods for the treatment of hematological neoplasms. Its successful application requires previous complete remission (CR) and exclusion of residual disease. MATERIAL/METHODS: This 31P MRS study reports data collected during several years of observation of nineteen patients. Duration of observation varied from 1 to 8.5 years from the time of diagnosis. Data collected after BMT covered periods ranging from 1.5 months to 6.5 years. RESULTS: 31P MRS spectra of normal sera showed four peaks, that at the lowest field due to Pi and the other three to phospholipids (PL), i.e. phosphatidylethanolamine + sphingomyelin (PE + SM), lysophosphatidylcholine (LPC), and phosphatidylcholine (PC). Long-term follow-up studies showed good correlation between the 31P MR spectral features of sera and disease response to therapy. In particular, at the time of diagnosis spectra showed significant decreases in the levels of all detected phospholipids (PC, LPC, PE and SM). These decreases, dependent upon disease severity, changed during chemotherapy according to the individual response of the patient. During chemotherapy and after BMT, the spectral profile changed in responding patients to resemble that of normal serum with the typical, higher peak intensities. CONCLUSIONS: Longitudinal analysis of in vitro 31P spectra of sera of patients with hematological malignancies represents a reliable method to evaluate the susceptibility of the neoplasm to a cytostatic treatment. The same approach seems to be useful in evaluating the effectiveness and outcome of bone marrow transplantation over time.