Cell counting of body fluids: comparison between three automated haematology analysers and the manual microscope method.

INTRODUCTION: Haematological analysis of body fluids (BF) specimens can provide clinicians with valuable diagnostic information because it can indicate one of several serious medical conditions. Although up to now the microscopic counting and the differentiation of WBC in a BF smear have been used as a reference. The introduction of semiautomated and automated methods of analysis has reduced interoperator variability and improved turnaround time and precision. The aim of our study was to evaluate the accuracy and the correlation between the three methods and with the reference method. METHODS: We examined 110 body fluid samples. Total counting of each sample has been conducted with all systems: Pentra DX120, ADVIA 2120 and XE-2100 and the manual method. RESULTS: We found statistically significant correlation between the data obtained in the ascitic and pleuric liquid but not in the cerebrospinal fluid. CONCLUSION: The introduction of automated method for BF analysis is more and more useful in the routine job of a laboratory analysis. It is therefore very important to evaluate the performance of the different automated haematology technologies, because there is a lack of literature in this field. The comparison between the Pentra DX 120, the other technologies and the manual counting showed instrumental overlapping capabilities.

Flow cytometric reticulocyte counting: a comparison between two methods.

The peripheral reticulocyte count is commonly used as an indicator of the erythropoietic activity of the bone marrow. Manual counting provides results with a high degree of inaccuracy and imprecision. Automation of counting is therefore needed. The increase in the number of methods available requires however that the results from the various methods agree with one another. The aim of our study was to evaluate the analytic performance of two automated hematology analyzers by a parallel study. We compared the analyzers between them and with manual counting. We enrolled in our study a total of 100 healthy subjects and an additional 80 patients affected by various hematological diseases. Difference between methods is statistically significant: the reference intervals of ADVIA2120 are higher than the Sysmex XE-2100. The correlation between methods and correlation with the microscopic method are excellent and statistically significant. In conclusion, we can affirm that total automation of reticulocyte counts represents a definite improvement over microscopic counts. This study confirms the diversity of the reference intervals still exists in the new automated hematology analyzers.

Erythrocyte and reticulocyte indices in iron deficiency in chronic kidney disease: comparison of two methods.

OBJECTIVE: Anaemia is a common complication of chronic kidney disease (CKD), particularly in dialysis patients. The recent European guidelines for anaemia treatment in CKD indicate the percentage of hypochromic red cells (%HYPO) and reticulocyte haemoglobin content (CHr) calculated by Siemens ADVIA haematology analysers as a useful tool indicating iron deficiency. The aim of this study was to evaluate the agreement between CHr and %HYPO parameters and the reticulocyte haemoglobin equivalent (RET-He) and red blood cell haemoglobin equivalent (RBC-He) calculated by the Sysmex XE-2100 haematology analyser in a cohort of 200 dialysis patients referred to the Nephrology Unit of our hospital. Furthermore, we evaluated a new index, the DF-Hypo XE, obtained from haemoglobin (Hb), haematocrit (Hct) and RET-He, provided by the Sysmex XE-2100, as a new potential marker of %HYPO in dialysed patients. MATERIAL AND METHODS: Blood samples collected in EDTA anticoagulant from 200 CKD patients receiving erythropoietin and iron to maintain haemoglobin level between 10 and 12 mg/dL were analysed on both the Siemens ADVIA 2120 and the Sysmex XE-2100 within 2 h of collection. RESULTS: There was good correlation between CHr and RET-He (r = 0.88; p<0.0001), %HYPO and DF-Hypo XE (r = 0.89; p<0.0001) and between RBC-He and CH (r = 0.96; p<0.0001), but there was a lower correlation, even though statistically significant, between RBC-He and %HYPO (r = -0.59; p<0.0001). The Altman-Bland analysis showed a very good level of agreement between CHr and RET-He (mean bias = 1.04 pg), %HYPO and DF-Hypo XE (mean bias = 1.73). Using a cut-off value of 29.4 pg for the RET-He and of 10.2 for the DF-Hypo XE, 15 out 17 patients with a CHr <29.0 pg and 9 out 11 patients with a %Hypo <10.0% were respectively correctly identified. CONCLUSIONS: Our study shows good correlation and agreement between CHr and RET-He and between %HYPO and DF-Hypo XE in evaluating CKD patients needing iron support.

Reference limits and behaviour of serum transferrin receptor in children 6-10 years of age.

Serum transferrin receptor (sTfR) originates mostly from erythroblasts and lesser from reticulocytes. The usefulness of sTfR has been implicated in several clinical situations, mainly as a marker of accelerated erythropoiesis or iron deficiency. The assessment of sTfR may be useful in the period of rapid growth during infancy, childhood and adolescence. We evaluated sTfR and the other quantitative and qualitative parameters of the erythropoiesis (Hb, MCV, CHr, Ret-He) and of the iron storage (serum ferritin, sTfR/ferritin index) in a total of 916 children aged 6-10 years. Children were divided into three groups: (A) healthy children, (B) with storage iron deficiency (serum ferritin < 12 microg/l) and (C) Beta trait carriers (HbA2 > 3.3). We determined reference intervals by sex and by age in healthy children. sTfR showed a slight but statistically significant age related increase but did not show significant sex differences. We compared sTfR and the other parameters investigated in the three groups of children. sTfR is not a decisive parameter that can be utilized alone in discriminating the border-line situations between normal and pathologic ones but can help in completing the panel of tests in iron deficiency and in thalassaemia Beta trait carriers.

Delayed decline of gamma-globin expression in infant age associated with the presence of Ggamma-158 (C-->T) polymorphism.

Persistent production of fetal hemoglobin (HbF) in adult has ameliorative effects on hemoglobinopathies and great efforts are currently made to achieve an exhaustive understanding of the molecular mechanisms of the switching in globin gene expression. One of the factors reported to be associated with the expression of fetal globin genes is the Xmn I Ggamma-158 polymorphism, although it is still unclear if it is involved in this mechanism either by itself or in strong linkage disequilibrium with other loci. Here, we report a novel effect of the Xmn I Ggamma-158 site that was found associated with a significant delayed decline of HbF production in infant age. The prolonged decay trend was enhanced when the Ggamma-158 C-->T substitution was co-inherited with a beta-thalassemic trait. Our observations reinforce the hypothesis that this region plays an important role in the expression of the gamma-globin genes and give new insights on the intriguing and still poorly understood mechanisms of globin gene expression switching.

Lipid profile in beta-thalassemia intermedia patients: correlation with erythroid bone marrow activity.

Lipid abnormalities, including low levels of all fractions of serum lipids, have been repeatedly reported in all phenotypes of beta-thalassemia. Unexpectedly, in more recent studies, the concentration of total cholesterol (TC) and high- and/or low-density lipoprotein cholesterol (HDL-C and LDL-C) has been found in beta-thalassemia intermedia (TI) patients even lower than in thalassemia major, without a clear explanation of pathophysiology of these findings. This lack of information prompted us to evaluate the plasma lipids and lipoproteins pattern in the TI patients followed in our department; the data were compared with those found in hereditary spherocytosis patients. Furthermore, in both groups of patients, the erythroid bone marrow activity was evaluated, utilizing the level of soluble transferrin receptors (sTfR) in the plasma. Both groups of patients showed similar lipid abnormalities (low-TC, HDL-C and LDL-C) and the same increase of sTfR, with significantly lower hemoglobin levels in TI patients. Data analysis of our study shows that the lipid profile in TI patients is not influenced by age, sex, liver injury, hemoglobin or ferritin levels; the higher erythroid bone marrow activity with the enhanced cholesterol consumption could be the dominant mechanism implicated in the lipid abnormalities of TI patients.

Hematologic values in healthy and small for gestational age newborns.

Many of the published reports of reference values in neonates are found in older medical literature. Recognition of abnormalities in blood cell morphology and hematologic parameters depend on well-established normative data; it is essential that each neonatal medical unit have its own reference ranges. We give the umbilical cord blood complete blood count reference values from 142 healthy, appropriate for gestational age (AGA) newborns and 58 small for gestational age (SGA) newborns (term and preterm). Our data, obtained by automated blood cell counter analysis of umbilical cord blood samples taken at birth, are comparable to other previously published data. The correlation between previous data and our reference data confirms that in term AGA newborns, values for red blood cells, hemoglobin, and hematocrit are higher and mean corpuscular volume values lower than in preterm AGA newborns. Also, we found that platelet levels are reduced in SGA newborns, in accordance with the literature. These findings further support the fact that preterm SGA infants are truly growth restricted, whereas term SGA infants are most likely small but otherwise healthy babies.

[Homozygous hemoglobin-E (Hb-EE) disease]. / Malattia da emoglobina E omozigote (Hb-EE).

The Authors report on a 16 year-old girl, of Cambodian descent, who was admitted to the hospital for hematuria. She showed a mild microcytic, hypochromic anemia with a normal iron balance; clinical examination was normal with neither pallor nor icterus nor splenomegaly; electrophoresis of hemoglobin yielded no hemoglobin A, a sligtly increased amount of HbF and a single band with a mobility similar to that of HbA2; the patient showed no evidence of overt increased hemolysis. With the DNA technology a final diagnosis of homozygous hemoglobin E was made. Hemoglobin E is the most common Hb variant among Southeast Asian populations. The Authors discuss on the benign nature of Hb-EE disease, pointing out that the presence of a single HbE gene in combination with that for beta-thalassemia leads generally to a disorder often comparable in severity to that of homozygous beta-thalassemia. With the recent migration of a high number of people from the countries, where HbE is extremely frequent, to the Western world (including Italy), this thalassemia syndrome is now a global health problem; therefore its knowledge is an important diagnostic challenge to all the experts involved in the care of thalassemic patients.

Selective enhancement of multipotential hematopoietic progenitors in vitro and in vivo by IL-20.

In a search for novel growth factors, we discovered that human interleukin-20 (IL-20) enhanced colony formation by CD34+ multipotential progenitors. IL-20 had no effect on erythroid, granulocyte-macrophage, or megakaryocyte progenitors. IL-20 transgenic mice increased the numbers and cell cycling of multipotential but not other progenitors. IL-20 administration to normal mice significantly increased only multipotential progenitor cells, demonstrating that IL-20 significantly influences hematopoiesis, with specificity toward multipotential progenitors. This is the first cytokine with such specificity identified.

Flow-cytometric analysis of erythrocytes and reticulocytes in congenital dyserythropoietic anaemia type II (CDA II): value in differential diagnosis with hereditary spherocytosis.

Congenital dyserythropoietic anaemia type II (CDA II) is the most common congenital dyserythropoietic anaemia. CDA II is frequently misdiagnosed as Hereditary Spherocytosis (HS) due to the presence of mild chronic haemolytic anaemia with splenomegaly, increased osmotic fragility, and presence of microspherocytes. Accurate diagnosis of CDA II is important to prevent severe iron overload. Erythrocyte and reticulocyte indices were assessed in 10 patients from six families with CDA II, 18 patients from eight families with HS, and 50 normal controls. Characteristic increases in distribution width were present in CDA II for cell volume (RDW, anisocytosis) and in HS for cell haemoglobin concentration (HDW, anisochromia), resulting in an RDW/HDW ratio which was significantly greater in CDA than HS (P < 0.0002). A cut-off value for RDW/HDW of 5.34 resulted in 89% sensitivity and 70% specificity in distinguishing CDA II from HS. Distribution width for cell haemoglobin content of reticulocytes (CHDWr) was characteristically increased in CDA II, resulting in a CHDW/CHDWr ratio significantly lower in CDA II than HS (P < 0.0002). A cut-off value of 0.98 provided 89% sensitivity and 80% specificity in distinguishing CDA II from HS. These differences in distribution widths of flow-cytometric parameters of reticulocytes and mature erythrocytes reflect the different pathogeneses of the two diseases and are helpful for the differential diagnosis of these two conditions.

[Long-term follow-up of chronic idiopathic thrombocytopenic purpura in children]. / "Follow-up" a lungo termine della porpora trombocitopenica cronica idiopatica in età pediatrica.

Idiopathic thrombocytopenic purpura (ITP) is a common acquired bleeding disorder in infancy and childhood. Most children rapidly improve, exhibiting a rise in platelet count to hemostatically normal levels within weeks to several months. Traditionally, chronic ITP is defined as persistence of thrombocytopenia (platelet count < 150 x 10(9)/L) for greater than 6 months. The Authors retrospectively evaluated 16 patients with chronic ITP, identified during a 12-year period of time in their Department of Pediatrics. The most important clinical and hematological parameters of patients were analyzed, including age at diagnosis, type and response to the initial treatment, number of multiple treatments, and duration of follow-up. At the last evaluation (december 1999) one patient was lost to the follow-up; one died of overwhelming postsplenectomy sepsis; four still require intermittent or chronic infusions of intravenous gamma-globulin; seven are in stable partial remission (PLT > 50 x 10(9)/L < 150 x 10(9)/L) and do not require any treatment; three are in complete remission (PLT > 150 x 10(9)/L). Finally, the Authors discuss of the natural history and management of this rare disease. Presently there are insufficient trial data to support evidence-based treatment guidelines in childhood chronic ITP and therefore it is reasonable to encourage future multicentre collaboration.

[Laurence-Moon-Bardet-Biedl syndrome. Physiopathological aspects and description of a clinical case]. / La sindrome di Laurence-Moon-Bardet-Biedl. Aspetti fisiopatologici e descrizione di un caso clinico.

After some general introductory remarks, a case of Laurence-Moon-Bardet-Biedl syndrome is reported. The main physiopathological aspects of the syndrome and some elements of differential diagnosis are discussed.