Evaluation assays measuring platelet kinetics in bone marrow and peripheral blood. An overview.

Platelets play an important role in haemostasis and thrombosis. For an understanding of the pathophysiology and treatment of thrombocytopenia, it is not sufficient to measure only the platelet count. Platelet kinetic parameters, such as platelet survival and turnover, might be useful because many thrombocytopenia related disorders result from the interaction between production, utilization or destruction, and sequestration of platelets. Therefore, measuring platelet turnover with radiolabelled platelets could be a sensitive and qualitative tool for clinicians. However, the method does not enjoy widespread use because it has some serious drawbacks, such as the problems associated with the manipulation of blood and platelets, and the use of radioactivity. Recently, other useful assays for measuring platelet fluxes have been described in the literature, including plasma thrombopoietin and glycocalicin. In this review, these new tests will be described, compared with the classical method using radiolabelled platelets, and finally evaluated for their usefulness in clinical practice.

Abnormal platelet adhesion on abdominal vessels in asymptomatic patients with paroxysmal nocturnal hemoglobinuria.

Platelet kinetic studies were performed in eight patients with paroxysmal nocturnal hemoglobinuria (PNH) but without symptoms or signs related to thrombosis. Indium-111 tropolonate labeled autologous platelets were used to determine mean platelet life and platelet production rate; total body images were also acquired. Three of seven patients with a thrombocytopenia showed a shortened mean platelet life, and the other five demonstrated a (nearly) normal platelet survival. Four of the seven patients with thrombocytopenia showed a decreased platelet production rate, but bone marrow cellularity was normo- to hypercellular. These observations suggest ineffective thrombopoiesis in 57% of the patients with PNH and thrombocytopenia. Total body imaging was performed during the platelet kinetic study in order to study organ uptake. Imaging of the abdominal vessels was demonstrated in all patients, suggesting enhanced adherence of platelets to the endothelium in patients with PNH. Since thrombosis of especially the abdominal vessels is a major cause of morbidity and mortality, the use of antithrombotic drugs might be considered despite the absence of abdominal complaints.

Effects of prednisone and splenectomy in patients with idiopathic thrombocytopenic purpura: only splenectomy induces a complete remission.

Idiopathic thrombocytopenic purpura (ITP) is a heterogeneous disease, whereby it is unclear if and in which way prednisone and splenectomy affect the platelet kinetics leading to a complete remission. To determine the effects of prednisone and splenectomy on the mean platelet life (MPL) and platelet production, platelet kinetic studies with Indium-111 tropolonate-labeled autologous platelets were performed in patients with ITP ( n=41). In 17 patients platelet kinetic studies were performed before and during prednisone treatment, and in 24 patients before and after splenectomy. MPL increased after prednisone therapy only in patients ( n=13) with a full recovery (FR, platelets >150 x 10(9)/l) and partial recovery (PR, 50 x 10(9)/l

Platelet kinetic studies in patients with idiopathic thrombocytopenic purpura.

PURPOSE: To determine the value in diagnosis and treatment of mean platelet life, platelet production, and major sites of platelet destruction in patients with idiopathic thrombocytopenic purpura (ITP). PATIENTS AND METHODS: Sternal or posterior superior iliac spine bone marrow aspiration was performed in 141 patients. Platelet kinetic studies with Indium-111 tropolonate labeled autologous platelets were utilized to determine platelet production. RESULTS: Two subgroups of patients could be defined. The first group (n = 81, 58%) had normal or increased platelet production and increased peripheral platelet destruction. These patients fulfilled the conventional criteria for ITP, including reduced platelet survival time (mean +/- SD, 1.6 +/- 1.4 days). Forty-eight (59%) of these patients had increased splenic sequestration and 30 (88%) of the 34 patients who underwent splenectomy had a complete or partial remission. The second group (n = 60, 42%) had decreased platelet production, with significantly greater platelet survival times (3.6 +/- 2 days, P <0.0001). In this group, the proportion of patients with complete or partial response to splenectomy (62%) was somewhat lower (P = 0.09). These patients mainly had ineffective platelet production in the bone marrow. CONCLUSIONS: Platelet kinetic studies suggest that ITP is a heterogeneous disease that comprises two subgroups. Further studies are needed to validate these findings and to determine their effect on the choice and outcome of therapy.

Rehydration following exercise: effects of administration of water versus an isotonic oral rehydration solution (ORS).

The effects of administering (1) 6L isotonic oral rehydration solution (ORS), similar in composition to plasma (except for an elevated potassium concentration) and with an osmotic skeleton and (2) 6L water (no osmotic skeleton), were evaluated in five thoroughbred horses following exercise-induced dehydration. The horses were exercised on a treadmill for 10 min at walk (1.7 m.s-1; approximately 15% VO2max), 40 min at trot (3.7 m.s-1; approximately 25% VO2max) and 10 min at walk (1.7 m.s-1; approximately 15% VO2max). Exercise was undertaken on a 3 degrees incline at 30 degrees C/80% RH. Solutions of water or ORS at 20 degrees C were administered by nasogastric tube over 60s 5 min following exercise. Mean weight loss following exercise was 9.2 +/- 1.7 kg (2.0 +/- 0.4% body weight; mean +/- SEM) with water and 9.2 +/- 1.1 kg (2.0 +/- 0.2% body weight) with ORS and was not different between treatments (P > 0.05). Water treatment resulted in a fall in plasma [Na+] (approximately 3 mmol.L-1) and C1- (1-2 mmol.L-1) concentrations by 30 min after administration and the effect persisted until the end of the study (300 min post fluids). There was little change in plasma total protein (TP) from that at the end of exercise, suggesting a failure of water to restore or maintain PV. In contrast, ORS administration resulted in a small increase in plasma [Na+] (1-2 mmol.L-1) and [Cl-] (2-3 mmol.L-1) with a corresponding decrease in plasma TP. By 120 min post ORS, plasma TP and PV were no longer significantly different from rest or pre-exercise (P > 0.05), whilst with water, TP was elevated (approximately 3-4 g.L-1) and PV reduced (approximately 4-5 mL.kg-1). Total urine output was not significantly different between water (1096 +/- 135 mL) and ORS (750 +/- 215 mL, P > 0.05). Estimates of expected plasma volume and electrolyte concentration changes as a result of either treatment compared well with measured changes of TP and PV. On the basis of calculated or measured changes, it was estimated that only 1L of water contributed to rehydration following exercise compared to 4L isotonic, plasma-like ORS. The administration of 6L ORS restored the PV deficit induced by exercise with minimal or no disturbance of plasma electrolyte concentrations. In contrast, water alone resulted in minimal improvement in PV. When fluid intake after periods of fluid loss, such as induced by exercise or transport, is not accompanied by food intake, the present study has clearly demonstrated that water alone is ineffective in promoting rehydration compared with an isotonic, plasma-like ORS.

Characterization of the erythropoiesis in myelodysplasia by means of ferrokinetic studies, in vitro erythroid colony formation and soluble transferrin receptor.

In refractory anemia (RA) and refractory anemia with ringed sideroblasts (RARS) a discrepancy is observed between the decreased in vitro erythroid colony formation and the normal or increased number of normoblasts in the bone marrow. To study the in vivo and in vitro erythropoiesis in more detail erythron transferrin uptake (ETU), soluble transferrin receptor (sTfR) and erythroid in vitro colony formation were performed in 24 patients with RA and five patients with RARS. These results were correlated with bone marrow morphology and transfusion dependency. Increased (mean, 124.9; range, 74-225 micromol/l blood/day) and normal (mean, 60.6; range, 50-71) ETU values were observed in 51% and 28% of the cases, whereas 21% of the cases demonstrated a diminished ETU value (mean, 35.8; range, 28-46), which correlated significantly with sTfR in cases with RA (P < 0.05, r = 0.64). A significant difference in ETU values was observed between RA (mean, 77.6; range, 28-189) and RARS (mean, 144.0; range, 59-225, P < 0.05). Most of the cases (73%) with increased ETU values showed an augmented percentage of erythroblasts in the bone marrow, which was inversely related with the serum Epo levels (P < 0.05, r = 0.51). However no correlation was found between the ETU values and the in vitro erythroid colony formation. Transfusion dependency was associated with normal to increased ETU levels (P < 0.05) and cytogenetic abnormalities (P < 0.05). These observations demonstrate that different patterns of defects can be observed in the erythropoiesis of RA and RARS patients whereby normal to increased ETU levels and the presence of cytogenetic abnormalities differentiate between cases of RA with ineffective erythropoiesis associated with regular transfusions and cases who are relatively transfusion independent.

Effects of administration of water versus an isotonic oral rehydration solution (ORS) at rest and changes during exercise and recovery.

The administration of 41 of an isotonic, plasma-like oral rehydration solution (ORS) with an osmotic skeleton and 41 of water (water; no osmotic skeleton), were evaluated in five thoroughbred horses. Solutions were administered by nasogastric tube 4 h after feeding. Uptake of deuterium, concentrations of plasma sodium, potassium, chloride, glucose, total protein and packed cell volume, pH, PCO2, HCO3-, total CO2, actual base excess, standard base excess, plasma volume and weight loss were assessed both at rest, and during and after exercise on a treadmill. Each horse underwent four experimental sessions (water-resting; ORS-resting; water-exercise; ORS-exercise). There was an indication of uptake of both water and ORS by 10 min post-administration. Based on the appearance of deuterium in plasma, there was no significant difference in the rate of uptake of water or ORS at rest, although there was a trend for the uptake of ORS to be slower than water during the exercise session. The mean decrease in total protein (TP, 3.0 g l-1) and the increase in plasma volume (PV, 4.6 ml kg-1) after administration of ORS at rest was greater (P < 0.05) than that of water (TP, 1.3 g l-1 and PV, -1.2 ml kg-1). There was no significant difference in TP or PV following administration of water or ORS during the exercise treatment. Exercise had little effect on plasma sodium concentration. The results confirmed that administration of 41 of isotonic, plasma-like ORS provided a much more distinct and durable contribution to the maintenance of plasma volume and circulation than administration of an equal amount of water at rest. Subsequent exercise may, however, mask some of the effects observed at rest.

Pyridoxin-responsive anaemia in a patient with a history of polycythaemia vera.

Pancytopenia in the course of polycythaemia vera (PV) following the proliferative and stable phase, ultimately leads to a spent phase characterized by extensive marrow fibrosis. We describe a patient with a history of PV and pancytopenia caused by myelodysplasia, before a genuine end stage myelofibrosis had occurred. The related anaemia was responsive to pyridoxin.

Normal values and reproducibility of left ventricular filling parameters by radionuclide angiography.

BACKGROUND: In physiologic situations age, heart rate (HR) and left ventricular ejection fraction (EF) may influence left ventricular filling rate. In this study, we determined normal values for radionuclide angiography (RNA) derived diastolic filling parameters, the correlations with age, HR and EF and their reproducibility. METHODS: The study was performed in 20 patients, 40-76 years old (mean 57), with normal findings at coronary angiography and left ventriculography. The first RNA was performed at rest (RNA1). Then, five minutes bicycle ergometry was performed and the patients were allowed five minutes rest before RNA was repeated (RNA2). From the left ventricular time activity curve we determined peak filling rate (PFR), time to peak filling rate (TPFR) and atrial contribution (AC) to ventricular filling. RESULTS: Values for PFR1 were 2.2 +/- 0.6 EDV/sec (PFR2 2.4 +/- 0.7 EDV/sec, r = 0.82), for TPFR1 198 +/- 22 msec (TPFR2 203 +/- 24 msec, r = 0.45) and for AC1 31 +/- 11% (AC2 31 +/- 10%, r = 0.72). The correlations of PFR and TPFR with age were statistically significant (respectively r = -0.68 and r = 0.48, P < 0.05). PFR was also influenced by HR and EF (resp. r = 0.51 and r = 0.50, P < 0.05). TPFR however was not influenced by HR and EF, whereas AC was positively correlated with HR (r = 0.79, P < 0.01). CONCLUSIONS: Radionuclide angiography is a reliable and reproducible method to assess parameters of diastolic left ventricular filling in individual patients. It may therefore be used to serially follow diastolic function. When used for interindividual comparison the dependency of RNA derived left ventricular filling parameters on age, HR and EF should however be considered.

111In-tropolonate labelled platelets; studies in normals and in patients with thrombocytopenia.

Human platelets were labelled with aqueous 111In-tropolonate in comparison with 111In-oxinate. In normals the labelling efficiency with 111In-tropolonate was higher (93% +/- 2%) than with 111In-oxinate (67% +/- 8%) (P less than 0.05). In cases of severe thrombocytopenia, lower labelling efficiencies were obtained. In six normals a mean platelet life of 9 days +/- 3 days and an initial recovery of 59% +/- 15% were obtained. In twelve patients with thrombocytopenia the mean platelet life was 4 days +/- 4 days and the initial recovery was 58% +/- 20%. The absolute uptake of radioactivity in spleen and in liver in both groups are reported.