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Simultaneous determination of chloroquine, amodiaquine and their metabolites in human plasma, red blood cells, whole blood and urine by column liquid chromatography.

A column liquid chromatographic method for the simultaneous determination of chloroquine, amodiaquine and their monodesethyl metabolite in human plasma, red blood cells, whole blood and urine is described. The drugs and internal standard were extracted as bases with methylene dichloride and then re-extracted into an acid aqueous phase. Separation was obtained using a reversed-phase column and a mobile phase of phosphate buffer (pH 3.0)-acetonitrile (88:12). The absorbance of the drugs was monitored at 340 nm with a sensitivity limit of 10 pmol/ml. No endogenous compound interfered at this wavelength. The mean overall recovery from each biological fluid was greater than 75%. This method can be applied to therapeutic, pharmacokinetic and epidemiological studies. The metabolism of these two amino-4-quinolines in humans is compared.

Amodiaquine↗

Effects of lost surface area on red blood cells and red blood cell survival in mice.

The effects of removing area from mouse red blood cells on the fate of the cells after reinfusion were investigated. When cells were made nearly spherical (by reducing cell area by approximately 35%) and then reinfused into the animal, most were cleared from the circulation within 1-2 h, although approximately 20% of the cells survived for 4 h or longer. When only 20% of the area was removed (leaving a 15% excess), more than 90% of the cells continued to circulate for 4 h. After reinfusion, the mean surface area of the surviving cells remained constant (73-75 microns2), but the mean volume decreased, from 56.6 +/- 2.1 to 19.1 +/- 1.5 microns3 (+/- SD of 5 replicates) over 4 h. These changes did not occur in cells suspended in plasma but not reinfused into the animal. Thus a loss of surface area results in a decrease in cell volume, as if to maintain a requisite degree of deformability. The results support the hypothesis that the increase in cell density associated with increasing cell age may be a consequence of surface area loss.

Animals↗

The societal unit cost of allogenic red blood cells and red blood cell transfusion in Canada.

BACKGROUND: There is a dearth of information about the cost of allogenic red blood cells (RBCs) and RBC transfusion in Canada in the aftermath of the Canadian blood system reorganization and the introduction of various safety measures. The unit cost of allogenic RBCs and RBC transfusion in Canada in 1994 was estimated at 152.17 US dollars. The objective of this study was to determine the unit cost of allogenic RBC transfusion in Canada from a societal perspective. STUDY DESIGN AND METHODS: A cost-structure analysis using the cost information from 2001 through 2002 was used. Costs of blood collection, production, distribution, delivery (hospital transfusion service processing and patient administration), transfusion reaction management, and opportunity cost of donor's time were included in the analysis. Canadian Blood Services and Héma-Québec supplied the data for collection, production, and distribution stages. Delivery and transfusion reaction costs were collected from eight hospitals across six Canadian provinces. In-patient costs were assessed for the intensive care unit, emergency, general medicine ward, and operating room. RESULTS: The aggregate mean societal unit cost of RBCs transfused on an inpatient basis in 2002 was 264.81 US dollars (95% confidence interval [CI], 256.29 dollars-275.65 dollars). The mean cost of blood collection, production, and distribution was 202.74 US dollars (95% CI, 199.63 dollars-204.31 dollars), the mean opportunity cost of donor time was 18.21 US dollars (95% CI, 17.11 dollars-21.63 dollars), the mean cost of hospital transfusion service processing was 16.65 US dollars (95% CI, 13.50 dollars-19.79 dollars), of RBC transfusion was 26.92 US dollars (95% CI, 25.33 dollars-28.52 dollars), and of transfusion reaction management was 0.29 US dollars(95% CI, 0.22 dollars-0.36 dollars). There were substantial variations in hospital transfusion service processing and RBC transfusion costs across hospitals. CONCLUSION: The societal unit cost of RBC transfusion has doubled since 1994 to 1995. Further increases in unit costs would be expected as additional safety measures are introduced. This will have important financial implications for treating patient populations that require a high level of RBC transfusions.

Blood Banks↗

Sister-chromatid exchange induction by sodium selenite: dependence on the presence of red blood cells or red blood cell lysate.

Sodium selenite (Na2SeO3) sister-chromatid exchange (SCE) induction was studied in both short-term and long-term cell cultures. The ability of Na2SeO3 to induce SCEs was found to depend on the culture conditions employed. Concentrations of Na2SeO3 (7.90 X 10(-6) M and greater) that produced elevated SCE frequencies in whole blood cultures resulted in control level SCE frequencies (6-8 SCEs/cell) in Ficoll-Hypaque--purified lymphocyte cultures. However, whole blood and purified lymphocyte cultures were equally sensitive to SCE induction by methyl methanesulfonate (MMS), ethyl methanesulfonate (EMS), and N-hydroxy-2-acetylaminofluorene (N-OH-AAF). Analysis of different whole blood components showed that the presence of red blood cells (RBCs), and specifically RBC lysate, was a prerequisite for Na2SeO3 SCE induction in purified lymphocyte cultures. The SCE frequencies of xeroderma pigmentosum (XP12RO) and normal human lymphoblastoid cell lines were also found to be unaffected by Na2SeO3 concentrations that produced elevated SCE frequencies in whole blood cultures. Incubation of these latter two cell types with Na2SeO3 and RBC lysate resulted in SCE frequencies comparable to those in Na2SeO3-exposed whole blood cultures.

Cell Line↗

Effect of red blood cells and red blood cell ghosts on reticuloendothelial system function.

Previous studies have shown that hepatic phagocytosis of red blood cell (RBC) stroma can depress reticuloendothelial system (RES) phagocytic function and increase susceptibility to shock. Since the RBC stroma used in these experiments contained substantial amounts of adherent hemoglobin, the present study was carried out to evaluate the role of the hepatic uptake of RBC membrane material on RES phagocytic function and susceptibility to endotoxin shock in rats. Neuraminidase-treated RBC which contained normal amounts of hemoglobin and RBC ghosts which were hemoglobin-free were used. Both preparations were removed from the circulation primarily by the liver. RES phagocytic function was depressed following the hepatic uptake of 29 X 10(8) neuraminidase-treated RBC and 26 X 10(8) RBC ghosts. RES uptake of neuraminidase-treated RBC was associated with an increase in susceptibility to endotoxin shock, but RBC ghosts did not affect shock susceptibility. Thus, RBC ghosts and intact RBC are equally effective in depressing RES phagocytic function, but RBC ghosts did not affect susceptibility to endotoxin shock.

Animals↗

Factors affecting the efficiency of collection of CD34-positive peripheral blood cells by a blood cell separator.

BACKGROUND: The yield of CD34-positive cells obtained from an apheresis procedure is determined, in part, by the efficiency of collection. Optimization of the efficiency of CD34-positive peripheral blood cell collection requires identification of predictive factors. STUDY DESIGN AND METHODS: Demographic, stem cell collection, mobilization, and disease-related measures from autologous and allogeneic donors undergoing 252 progenitor cell apheresis procedures were retrospectively reviewed. Statistical relationships between CD34 collection efficiency and the various measures were determined by correlation and multiple linear regression analysis. RESULTS: CD34 collection efficiency inversely correlated with the peripheral white cell count, hematocrit, and serum albumin concentration (R2 = 0.29). White cell count was the single best predictor of CD34 efficiency (R2 = 0.19). Donor groups with cytopenias (patients vs. normal donors; increased cycles of prior chemotherapy; bone marrow involvement; chemotherapy plus growth factor mobilization) had higher collection efficiencies. Only 29 percent of the variability in the data could be attributed to white cell count, hematocrit, and albumin concentration. The majority of the remaining variability was due to unexplained differences between donors. CONCLUSION: CD34 collection efficiencies show considerable variation. Higher peripheral white cell counts, hematocrits, and/or albumin concentrations result in decreased CD34 collection efficiency, but most of the variability in the data is not accounted for by these three factors.

Adolescent↗

[Difference of blood cell counts with reference blood cell counters in different makers: Part II].

This is the second report of evaluation on the difference of blood cell counting among different automated blood cell counters in Japan. We tested reference blood cell counters of 6 different companies: Coulter, Sysmex, Bayer-Sankyo, Nihon Kohden, Horiba and Dainabot. Forty ml of whole blood were taken from 3 healthy persons and EDTA-2K anticoagulated blood samples (Sample 1, 2 and 3) were sent to each company to determine blood cell counts with a reference automated counter. As a result, the following items showed more than 10% difference among makers: RBC between Dainabot and Horiba in Sample 3, hematocrit values between Coulter and Dainabot in Sample 2, WBC between Nihon Kohden and each of three makers (Sysmex, Horiba and Dainabot) in all 3 samples and that between Bayer-Sankyo and each of two makers (Sysmex and Horiba) in Sample 3 and platelet count between Dainabot and each of 3 makers (Bayer-Sankyo, Nihon Kohden and Horiba) in all 3 samples. The following items showed difference between 5 and 10%: MCV between Coulter and each of two makers (Bayer-Sankyo and Horiba), WBC between each of two makers (Coulter and Nihon Kohden) and each of other 4 makers, and platelet count between each of two makers (Nihon Kohden and Horiba) and each of 3 makers (Coulter, Sysmex and Bayer-Sankyo). Recently Japanese Committee for Clinical Laboratory Standards proposed minimum clinical allowance of blood cell count as follows: hemoglobin 3%, RBC 4%, MCV 4%, WBC 7% and platelet count 10%. It is suggested that all of the items showing the difference more than above allowance among makers should be improved for clinical use to have good external quality control in blood cell counting by automated instruments.

Blood Cell Count↗

Prevention of immunization to D+ red blood cells with red blood cell exchange and intravenous Rh immune globulin.

BACKGROUND: Although young women who are D- occasionally receive unintentional transfusions with D+ red blood cells (RBCs), there are little data to assist with management of such an event. Two cases of D- girls transfused with D+ RBCs are reported. In an effort to prevent formation of anti-D, RBC exchange followed by administration of intravenous (IV) Rh immune globulin (RhIg) was used. CASE REPORTS: Patient 1, a 56-kg, 16-year-old D- girl, was involved in a motor vehicle crash. She received 4 units of Group O uncrossmatched D+ RBCs. Thirty-six hours after admission, she underwent RBC exchange with 10 units of D- RBCs, followed by a total of 2718 microg of IV RhIg over 32 hours. Six months later, her antibody screen was negative. Patient 2, a 39-kg, 10-year-old D- girl with aplastic anemia, received 1 unit of D+ RBCs. She underwent RBC exchange on the same day with 5 units of D- RBCs, followed by a total of 900 microg of IV RhIg over 8 hours. Six months later her antibody screen was negative. CONCLUSION: RBC exchange followed by a calculated dose of IV RhIg was successful in preventing allo-immunization to D. Several small studies suggest that both trauma and hematology patients may be less capable of becoming immunized with the transfusion of D+ blood components. Until these findings are more clearly defined, there will be times when prevention of immunization of any D- girl is desired. RBC exchange followed by RhIg appears to be one way to achieve this goal.

Adolescent↗

High in-hospital mortality of intensive care patients with nucleated red blood cells in blood.

The detection of nucleated red blood cells (NRBCs) in blood of patients suffering from a variety of severe diseases is known to be highly associated with increased mortality. Blood analyzers to routinely measure NRBC concentrations are now available. However, the diagnostic and prognostic significance of this parameter for intensive care patients has not been evaluated. Using a Sysmex XE-2100 analyzer, NRBC concentrations were determined in blood samples from 421 patients treated in intensive care units (general and accident surgery, cardiothoracic surgery, and internal medicine) of a university hospital. NRBCs were found at least once in 19.2% of all patients. The mortality of NRBC-positive patients (n=81) was 42.0% (n=34); this was significantly higher (p<0.001) than the mortality of NRBC-negative patients (5.9%, n=340). The NRBC concentration was 115+/-4x10(6)/l (median 40x10(6)/l; range 20-2930x10(6)/l) at initial detection of NRBCs in the blood. Mortality increased with increasing NRBC concentration and increasing frequency of occurrence. With regard to in-hospital mortality, NRBCs in blood showed sensitivity and specificity of 63.0% and 87.2%, respectively. The detection of NRBCs is highly predictive of death, the odds ratio after adjustment for other laboratory prognostic indicators being 1.01 (p<0.01) for each increase in the NRBC concentration of +1x10(6)/l. NRBCs were detected for the first time, on average, 13 days (median 8 days) before death. The routine analysis of NRBCs in blood is of high prognostic power with regard to in-hospital mortality of critically ill patients. Therefore, this parameter may serve as an early indicator for patients at increased mortality risk.

Critical Care↗

Quantitation, mobilization, and clinical use of long-term culture-initiating cells in blood cell autografts.

Mobilized peripheral blood is increasingly used as a source of cells to provide hematopoietic reconstitution in marrow ablated patients. There is controversy as to how the engraftment potential of these preparations should be evaluated. The long-term culture-initiating cell is a very primitive hematopoietic with unique phenotypic feature. The potential role and significance of these cells in peripheral blood cell autografts is discussed.

Animals↗

A comparison of washed red blood cells versus packed red blood cells (AS-1) for cardiopulmonary bypass prime and their effects on blood glucose concentration in children.

The effects on blood glucose concentrations of packed red blood cells (AS-1) (group I) versus washed red blood cells (group II) for cardiopulmonary bypass prime were compared in 20 infants weighing less than 10 kg undergoing cardiac surgical procedures. All patients were anesthetized with N2O/O2/isoflurane/fentanyl and received lactated Ringer's solution prior to bypass. Blood glucose concentrations prior to bypass were 85 +/- 15 mg/dl (mean +/- SD) in group I and 81 +/- 14 mg/dl in group II. Blood glucose concentrations were 210 +/- 21 mg/dl versus 78 +/- 14 mg/dl (P less than 0.001) 10 min after initiation of bypass, 241 +/- 48 mg/dl versus 107 +/- 28 mg/dl (P less than 0.001) prior to separation from bypass, and 214 +/- 52 mg/dl versus 97 +/- 19 mg/dl (P less than 0.001) after protamine administration in group I and group II, respectively. The use of washed red blood cells for cardiopulmonary bypass priming solution in infants significantly attenuates the increase in blood glucose concentration otherwise observed during cardiopulmonary bypass.

Adenine↗

Influence of blood cells and blood flow on venous endothelium.

The principal emphasis in the past has been on the physical characteristics of the vein wall and to some extent the flow characteristics of the blood it contains. The interaction between the blood cells and the venous endothelium has been largely neglected until recently. This brief review summarises the secretion by the endothelium of such important substances as fibrinolytic agents, antiplatelet agents and venodilators. Many of these secretory activities are modulated by the wall shear stress which in turn is related to the rheological properties of the blood. The rheological properties of bulk venous blood have been shown to be significantly abnormal in patients with venous hypertension. Such haemodynamic and haemorheological alterations also play an important role in the interaction between the formed elements of blood and the vessel wall. Most recently this has assumed the importance in relation to the margination and adhesion of white cells to the endothelium and the subsequent activation of the white cells. Interaction between the circulatory blood and the venous endothelium probably plays an important role in the pathophysiology of both simple varicose veins and the more serious complications of venous disease such as thrombosis.

Cell Adhesion↗

Induction of trunk lateral cells, the blood cell precursors, during ascidian embryogenesis.

The tadpole larvae of the ascidian Halocynthia roretzi have trunk lateral cells (TLCs) in their trunk. TLCs give rise to adult blood cells after metamorphosis. TLCs are exclusively derived from the A7.6 cell pair of 64-cell embryos. When prospective TLC blastomeres were isolated from embryos before the 16-cell stage, they failed to express TLC-specific antigen, a molecular indication of differentiation of TLCs. Isolates after the 32-cell stage, however, autonomously expressed the antigen. Results of experiments involving coisolation and recombination of blastomeres at the 16-cell stage showed that the inductive influence emanating from cells of animal hemisphere (presumptive epidermis blastomeres) is required for TLC formation. The inductive interaction takes place at the 16-cell stage, two cell cycles before the developmental fate becomes exclusively restricted to TLC formation. The inducing activity is distributed widely in animal hemisphere. By contrast, only presumptive TLC blastomeres have competence to be induced to form TLCs.

Animals↗